2003 JOURNAL PAPER Summaries based on PubMed

(2003). "[In Process Citation]." Morfologiia 124(4): 47-50.
 An expression of high molecular component of neurofilament triplet NF200 (marker of neurons forming A-fibers) and binding of isolectin B4 (IB4) was examined immunohistochemically in LIV-LV dorsal root ganglia (DRG) neurons after ligation or transection of the sciatic nerve in rat. NF200 immunoreactivity was detected in 15% of all neurons in DRG of intact rats. Ligation of sciatic nerve caused a two-fold decrease in number of NF200-positive neurons by 90th day after nerve injury, however in animals treated with peripheral nerve regeneration stimulator xymedon the number of surviving NF200-positive neurons was increased by 50.7% as compared with control group (nerve ligation without treatment). In DRG of intact rats 23.6% of neurons showed IB4 binding. Of the DRG neurons 2.6% were labeled by IB4 at 30th day after ligation of the nerve. At 90th day after ligation no IB4-positive neurons were revealed in DRG of untreated rats, while xymedon treatment was shown to result in more than 8-fold increase in the number of surviving IB4-positive neurons. IB4-positive neurons have greater probability of entering the posttraumatic apoptosis. After nerve ligation the survival of NF200- and IB4-positive neurons was less than that one following nerve transection, suggesting that axon lengthening could be a the factor supporting neuronal survival. Pyrimidine derivative xymedon promoted the survival of neurons in both subpopulations with predominant effect on IB4-positive neurons.

(2003). "[In Process Citation]." Morfologiia 124(4): 38-40.
 In the present study the immunohistochemical localization of proline-rich peptide, so called galarmin, was examined in the brain structures of intact and galarmin-treated rats. Galarmin (a fragment of neurophysin II C-end glycopeptide) was isolated by A.A. Galoyan and coworkers in 1997, from the neurosecretory granules of bovine neurohypophysis, produced by the hypothalamic magnocellular nuclei. In intact rats galarmin-immunoreactive neurons and nerve fibers were widely distributed in the central nervous system. Single intramuscular injection of galarmin to the rats resulted in the increase of both galarmin-immunoreactivity and the number of galarmin-immunoreactive nerve cells, fibers and capillaries. In control experiments where the antisera against the fragment of immunophilin (a receptor of immunosuppressor macrolide FK-506) and the pancreatic neuropeptide Y were used as the primary antibodies, the significant increase of neuropeptide Y-immunoreactive nerve fibers and immunophilin-positive lymphocytes was revealed in galarmin-treated rats. Based on these results and the data on the motoneurons regeneration in the spinal cord hemisectioned rats given galarmin daily for 3 weeks, galarmin has been suggested to act as an immunomodulator, neurotransmitter and neuroregulator.

(2003). "Retinal cell rescue: stem cells, remodelling, neuroprotection, apoptosis and replacement of retinal neurons. Proceedings of the 6th Annual Vision Research Conference. Ft. Lauderdale, Florida, USA. May 3-4, 2002." Vision Res 43(8): 857-956.
 
(2003). "Summaries for patients. Nerve growth factor improves healing of pressure ulcers of the foot." Ann Intern Med 139(8): I10.
 
(2003). "[In Process Citation]." Vestn Khir Im I I Grek 162(4): 30-3.
 Experiments in 26 dogs were carried out in order to analyze the quality of comparing the nerve fiber bundles, morphometrical and functional parameters of regeneration after the restoration of the nerve with a standard epineural microsurgical suture and by an original method including the coaptation of the freshened ends by a relaxation transneural suture followed by epiperineural microneurorrhaphy. Five variants of the comparison of the nerve fiber bundles in the suture zone are described. The results of expert assessments and computed morphometry allowed to register higher indices of regeneration and differentiation of nerve fibers.

Aagaard, B. D., D. A. Lazar, et al. (2003). "High-resolution magnetic resonance imaging is a noninvasive method of observing injury and recovery in the peripheral nervous system." Neurosurgery 53(1): 199-203; discussion 203-4.
 OBJECTIVE: Noninvasive observation of degenerating and regenerating peripheral nerves could improve the diagnosis and treatment of nerve injuries. We constructed a novel phased-array radiofrequency coil to permit magnetic resonance imaging (MRI) observation of the sciatic nerve and its target muscles in rats after injury. METHODS: Adult male Lewis rats underwent either crushing (n = 18) or cutting and capping (n = 17) of their right sciatic nerves and then underwent serial MRI. Serial gait track analysis was performed to assess behavioral recovery. Animals from both groups were killed at several time points for histological evaluation of the nerves, with axon counting. RESULTS: Crushed sciatic nerves demonstrated increased T2-weighted signals, followed by signal normalization as axonal regeneration and behavioral recovery occurred. Cut sciatic nerves prevented from regenerating displayed a prolonged phase of increased signal intensity. Acutely denervated muscles exhibited hyperintense T2-weighted signals, which normalized with reinnervation and behavioral recovery. Chronically denervated muscles demonstrated persistently increased T2-weighted signals and atrophy. CONCLUSION: In this study, we demonstrated the ability of MRI to noninvasively monitor injury and recovery in the peripheral nervous system, by demonstrating changes in nerve and muscle that correlated with histological and behavioral evidence of axonal degeneration and regeneration. This study demonstrates the potential of MRI to distinguish traumatic peripheral nerve injuries that recover through axonal regeneration (i.e., axonotmetic grade) from those that do not and therefore require surgical repair (i.e., neurotmetic grade). This diagnostic modality could improve treatment by providing earlier and more accurate diagnoses of nerve damage, as well as reducing the need for exploratory surgery.

Abe, I., A. Tsujino, et al. (2003). "Effect of the rate of prestretching a peripheral nerve on regeneration potential after transection and repair." J Orthop Sci 8(5): 693-9.
 We tested the influence of nerve stretch injury on nerve regeneration after cutting and suturing. An external fixator was used to lengthen the femur, and consequently the sciatic nerve, progressively, by 3 mm/day (5%/day: group I) or 5 mm/day (9%/day: group II). In both groups the total lengthening was 15 mm, corresponding to approximately 23%-28% elongation of the sciatic nerve. Seven days after initiating nerve lengthening, the external fixator was removed and the nerve was transected and sutured. At 2, 4, 6, or 8 weeks after this suturing, semithin sections were prepared from the sciatic and tibial nerves. Although regeneration of nerve fibers was observed beginning at 2 weeks in all groups, nerve regeneration showed a delay only in group II at 6 weeks, when group I showed nerve regeneration as good as in the unstretched control group. Differences in nerve regeneration after nerve lengthening at different rates were considered the result of mechanical nerve injury, nerve degeneration, ischemia, and fibrosis, all made worse with more rapid lengthening. Nerve lengthening at a more moderate rate (3 mm/day) did not appear to compromise regeneration.

Abe, S., I. Mizusawa, et al. (2003). "Changes in mRNA expression patterns for cytokines in blood leukocytes of a rat tourniquet model." Leg Med (Tokyo) 5 Suppl 1: S275-7.
 We examined changes in mRNA expression patterns for proinflammatory cytokines and growth factors in blood samples after application of a tourniquet to the rat hind limb. Slight upregulations of interferon (IFN)-gamma, macrophage colony-stimulating factor (M-CSF) and transforming growth factor (TGF)-beta1 mRNA began at 2h after tourniquet application and were short-lived. The levels of activating transcription factor (ATF)-3, a stress-inducible gene, had increased at 1h after tourniquet application. No significant expression of interleukin (IL)-6 mRNA was observed in most samples. There were no significant temporal changes in the levels of IL-1beta, cardiotrophin (CT)-1 mRNA compared to the control levels, but, downregulation of gp130, a receptor of the IL-6 family, began at 1h after tourniquet application. Nerve growth factor (NGF) mRNA gradually increased and reached a significantly high level at 4h after application of the tourniquet. Gene expression induction in blood leukocytes occurred soon after application of the tourniquet and was short-lived. The transient mRNA expressions probably trigger secondary events that may be beneficial to wound repair and regeneration.

Abrams, C. K., M. Freidin, et al. (2003). "Pathogenesis of X-linked Charcot-Marie-Tooth disease: differential effects of two mutations in connexin 32." J Neurosci 23(33): 10548-58.
 X-linked Charcot-Marie-Tooth disease is an inherited peripheral neuropathy arising in patients with mutations in the gene encoding connexin 32 (Cx32). Cx32 is expressed at the paranodes and Schmidt-Lantermann incisures of myelinating Schwann cells in which it is believed to form a reflexive pathway between the abaxonal and adaxonal cytoplasmic domains. Patients with the Val181Ala (V181A) mutation have a severe peripheral neuropathy. Experiments using a nude mouse xenograft system show that Schwann cells expressing only this mutant form of Cx32 are profoundly impaired in their ability to support the earliest stages of regeneration of myelinated fibers. Coupling between paired Xenopus oocytes expressing V181A is reduced compared with the coupling between oocytes expressing wild-type human Cx32 (32WT), and protein levels assayed by Western blot are substantially lower. Immunocytochemisty shows that Neuro2a cells expressing the V181A mutant have very few gap junction plaques compared with cells expressing 32WT; Cx32 protein levels are lower in these cells than in those expressing 32WT. Because failure of normal regeneration is evident before formation of myelin, loss of function of Cx32 may impact on the function of precursors of the myelinating Schwann cell before the formation of the hypothesized reflexive pathway. The Glu102Gly (E102G) mutation leads to a milder phenotype. Early regeneration is normal in grafts with Schwann cells expressing the E102G mutant. The only abnormality detected in the behavior of its channel is increased sensitivity to acidification-induced closure, a property that may lead to reduced gap junction coupling during periods of metabolic stress. This restricted functional abnormality may explain the relatively mild phenotype seen in the xenograft model and in E102G patients.

Ackermann, P. W., J. Li, et al. (2003). "Neuronal plasticity in relation to nociception and healing of rat achilles tendon." J Orthop Res 21(3): 432-41.
 Nerve regeneration and the occurrence of three neuropeptides; i.e. substance P (SP), calcitonin gene related peptide (CGRP) and galanin (GAL), were studied during healing of tendon rupture in the rat by semi-quantitative immunohistochemistry. The neuronal findings were related to nociception as assessed by hindpaw withdrawal latencies at thermal and mechanical tests.Experimental rupture of rat Achilles tendon--normally devoid of nerves--elicited extensive nerve ingrowth into the rupture site in the early phase of healing followed by almost complete fiber disappearance (weeks 12-16). The ingrowth of SP and CGRP positive fibers, seen already at weeks 1-2, was associated with increased nociception. Subsequently, the occurrence of GAL positive fibers at weeks 4-6 was associated with decreased nociception. An even stronger relationship to nociception during healing was observed when the rate of change in neuropeptide expression instead of the expression in absolute terms was considered, according to the "cascade" formula of SP(')+CGRP(')-GAL(').It may prove that the observed temporal occurrence of different neuropeptides reflects a role of the peripheral nervous system in regulating synchronously nociception and healing.

Adams, D. S., B. Hasson, et al. (2003). "A peptide fragment of ependymin neurotrophic factor uses protein kinase C and the mitogen-activated protein kinase pathway to activate c-Jun N-terminal kinase and a functional AP-1 containing c-Jun and c-Fos proteins in mouse NB2a cells." J Neurosci Res 72(3): 405-16.
 Ependymin (EPN) is a goldfish brain neurotrophic factor previously shown to function in a variety of cellular events related to long-term memory formation and neuronal regeneration. CMX-8933, an 8-amino-acid synthetic peptide fragment of EPN, was designed for aiding an investigation of the biological properties of this glycoprotein. We reported from previous studies that treatment of mouse neuroblastoma (NB2a) cultures with CMX-8933 promotes activation of transcription factor AP-1, a characteristic previously associated with the following full-length neurotrophic factors: nerve growth factor, neurotropin-3, and brain-derived neurotrophic factor. The CMX-8933-activated AP-1 specifically bound an AP-1 consensus probe and appeared to contain c-Jun and c-Fos protein components in antibody supershift experiments. Because AP-1 influences a variety of positive and negative cellular processes, determined in part by its exact protein composition and mechanism of activation, we extended these initial AP-1 observations in the current study to confirm the identity of the CMX-8933-activated c-Jun and c-Fos components. CMX-8933 increases the enzymatic activity of c-Jun N-terminal kinase (JNK), increases the phosphorylation of JNK and c-Jun proteins, and increases the cellular titers of c-Jun and c-Fos mRNAs. Furthermore, the AP-1 activated by CMX-8933 is functional, insofar as it transactivates both synthetic and natural AP-1-dependent reporter plasmids. Inhibition studies indicate that activation of the 8933-induced AP-1 occurs via the mitogen-activated protein kinase pathway. These data are in agreement with the recently proposed model for the conversion of short- to long-term synaptic plasticity and memory, in which a JNK-activated transcription factor AP-1, containing c-Jun and c-Fos components, functions at the top of a hierarchy of transcription factors known to regulate long-term neural plasticity.

Adanali, G., M. Verdi, et al. (2003). "Effects of hyaluronic acid-carboxymethylcellulose membrane on extraneural adhesion formation and peripheral nerve regeneration." J Reconstr Microsurg 19(1): 29-36.
 The goal of nerve repair in the peripheral nervous system is to increase the number of axons passing from proximal to distal stump, and to enable the regenerated axons to reach the end organ as soon as possible. In the present study, the effect of the membrane formed by a mixture of hyaluronic acid and carboxymethylcellulose (HA-CMC) on nerve regeneration and perineurial scar formation was investigated. Eighteen New Zealand rabbits were allocated into control (n = 9) and experimental groups (n = 9). In the control group, conventional nerve repair was carried out following the transection of the sciatic nerve, while in the experimental group, following repair of the nerve, the repair line was covered by HA-CMC membrane extending 1 cm beyond the distal and proximal ends. Nerve regeneration and extraneurial adhesion formation were compared between the two groups 3 months later. It was observed that adhesion in the surrounding tissues was significantly less in the experimental group than in the control group. Furthermore, morphometric analysis of specimens obtained from the distal parts of nerves showed that the number of axons with myelin was higher in the experimental group than in the control group, with a statistically significant difference. Histologic sections obtained from the nerve repair line demonstrated that extraneural and intraneural fibrosis was significantly lower in the experimental group. It was concluded that HA-CMC membrane had a favorable effect on nerve regeneration, as well as extraneural scar formation, encouraging the clinical application of HA-CMC following nerve injuries.

Adams, M. (2003). "Third annual meeting of the California Spinal Cord Injury/Neural Regeneration Consortium March 20-21, 2002, the Reeve-Irvine Research Center of the University of California, Irvine, USA." Spinal Cord 41(2): 134-6.
 
Adani, R., I. Marcoccio, et al. (2003). "Flap coverage of dorsum of hand associated with extensor tendons injuries: A completely vascularized single-stage reconstruction." Microsurgery 23(1): 32-9.
 This study reports results in 12 patients treated with "completely vascularized single-stage approaches," so defined because skin, tendon, and nerve are transferred as a compound flap, and all are vascularized. A free dorsalis pedis cutaneotendinous flap was used in 7 patients, while a radial forearm cutaneotendinous island flap was transposed in 5 patients. A dorsalis pedis flap provides four vascularized extensor tendons (extensor digitorum comunis tendons), and the radial artery flap permits the inclusion of one completely vascularized tendon (palmaris longus) and two "strips" of vascularized tendons (flexor carpi radialis and brachioradialis). The flaps survived in all cases, and the transferred tendons were functioning well. The dorsalis pedis flap can be employed in the reconstruction of cutaneotendinous defects of the dorsum of the hand which require the use of three or four tendons grafts. We suggest the use of forearm cutaneotendinous flaps in cases of reconstruction of one or two extensor tendons. The "completely vascularized single-stage reconstruction" avoids prolonged hospitalization and results in a rapid restoration of near-normal function and appearance of the hand.

Agbulut, O., M. L. Menot, et al. (2003). "Temporal patterns of bone marrow cell differentiation following transplantation in doxorubicin-induced cardiomyopathy." Cardiovasc Res 58(2): 451-9.
 OBJECTIVE: Recent studies have suggested benefits of bone marrow cell transplantation for the regeneration of ischemic cardiac tissue. To extend the potential of cell transplantation, we assessed this treatment in a mouse model of acute nonischemic doxorubicin-induced cardiomyopathy. METHODS: To allow detection of engrafted cells, we used transgenic mice expressing the nuclear-located LacZ under the control of either desmin or vimentin promoters, which identify muscle lineage and mesenchymal cells, respectively. All transplanted cells were also labeled with the fluorescent dye DIL. One week after the administration of doxorubicin (15 mg/kg), mice were intramyocardially injected with either allogeneic unpurified bone marrow cells (6 x 10(6) in 30 microl, n=59) or purified sca-1(pos) cells (4 x 10(5) in 30 microl, n=22). In parallel, control normal mice received only unpurified bone marrow cells (n=28). Hearts were harvested at serial intervals until 2 weeks after transplantation and analyzed by immunohistochemistry to assess the degree of engraftment and transplanted cell differentiation. RESULTS: In control mice, no differentiation of bone marrow cells was detected. In contrast, unpurified bone marrow cells grafted into diseased myocardium featured two successive phases of cell differentiation. The first yielded cells with a mesenchymal phenotype (44.1+/-10.1 cells/3 x 10(-2) mm(3) at 2 days), was transient and lasted 1 week. The second phase was characterized by cells with a muscular phenotype detected in a small number of cells (5.6+/-2.3 cells/3 x 10(-2) mm(3) at 7 days). Two weeks after transplantation, some of these cells appeared phenotypically close to cardiomyocytes, as evidenced by morphology and positive staining for myosin binding protein C, vinculin and myosin heavy chain. In sca-1(pos) hematopoietic progenitor grafted mice hearts, no transdifferentiation into cardiac cells was detected at any time point. CONCLUSION: These data support the hypothesis of the potential for a myogenic differentiation of bone marrow cells following engraftment in a nonischemic model of global cardiomyopathy. Bone marrow-derived cells amenable to cardiac differentiation are present in total unpurified bone marrow but not in the sca-1(pos) hematopoietic progenitor cell population. However, the very small number of transdifferentiated cells raises concerns over their functional efficacy.

Ahmed, Z., S. Underwood, et al. (2003). "Nerve guide material made from fibronectin: assessment of in vitro properties." Tissue Eng 9(2): 219-31.
 We have previously used orientated mats of fibronectin as conduits to repair short gaps in peripheral nerves. Here we describe the in vitro properties of a new material in the form of large cables produced from a fibronectin-enriched solution with potential as a conduit for longer nerve defects. Large cables of fibronectin were made up to 14 cm long x 1.5 cm in diameter. When freeze dried, scanning electron microscopy revealed a predominant fiber orientation. Dried cables hydrated rapidly to 1.6 and 4.8 times their original length and diameter, respectively. Once hydrated these cables had pores that ranged from 10 to 100 microm through which Schwann cells and fibroblasts were able to grow in vitro and align with the axis of the fibrils by contact guidance. Furthermore, the porosity of the cable was enhanced by the natural dissolution of protein over a 3-week duration in culture with cells, such that 50- to 200-microm pores were observed. This study suggests that large fibronectin cables are a suitable alternative to the original fibronectin mats to guide components of the peripheral nerves and so to act as conduits with potential use in guiding regeneration across long nerve defects.

Akassoglou, K., P. Akpinar, et al. (2003). "Fibrin is a regulator of Schwann cell migration after sciatic nerve injury in mice." Neurosci Lett 338(3): 185-8.
 Fibrin, derived from the blood protein fibrinogen, is deposited in the sciatic nerve after injury and retards functional regeneration. Since Schwann cell migration is critical for remyelination of injured nerves, we investigated the effects of fibrin in this process. In vivo experiments showed that fibrin co-localizes with fibronectin deposition in the injured sciatic nerve. In vitro migration assays demonstrated that fibrin alone is not a permissive substrate for Schwann cell migration. Furthermore, migration assays of Schwann cells on mixed fibrin/fibronectin substrates showed that fibrin has a dose dependent inhibitory effect on Schwann cell migration on fibronectin. Our results show that fibrin, deposited in the sciatic nerve after injury, changes the composition of the extracellular matrix and inhibits Schwann cell migration. This negative effect of fibrin should be considered in the therapeutic application of biomaterials based on fibrin matrices.

Allegretto, M., M. Morrison, et al. (2003). "Selective denervation: reinnervation for the control of adductor spasmodic dysphonia." J Otolaryngol 32(3): 185-9.
 OBJECTIVES: The objective of this study was to evaluate the efficacy of a new surgical procedure for adductor spasmodic dysphonia (AddSD). This surgery involves the bilateral selective division of the adductor branches of the recurrent laryngeal nerves with immediate reinnervation of the distal nerve trunks with branches of the ansa cervicalis (selective denervation-reinnervation). METHODS: Our first six patients to undergo this procedure were enrolled in the study. All patients suffered from AddSD and had previously received botulinum toxin A (Botox, Allergen, Markham, ON) therapy. Patients were recorded preoperatively and all underwent the same surgical procedure performed by the same lead surgeon. All patients were surveyed postoperatively and then re-recorded. Expert and untrained judges undertook perceptual evaluation of voice quality. Voice samples were also objectively evaluated for aphonic voice breaks. RESULTS: No major surgical complications were noted. Patient satisfaction was excellent, and five of the six patients no longer require botulinum toxin therapy. In five of the six patients, the majority of untrained and expert listeners perceived the postoperative voice to be superior. Objectively, the rate of aphonic voice breaks was also reduced in five of the six patients.

Alsancak, S. (2003). "Splint satisfaction in the treatment of traumatic radial nerve injuries." Prosthet Orthot Int 27(2): 139-45.
 Between the years 1986-2001, 135 patients who have had traumatic radial nerve injury, were supplied with dynamic wrist-hand orthoses (WHOs). The aim of this study is to examine the applied splints, considering aspects such as apperance, practicality of use, function, comfort and endurance. The author's assessments show that the biggest problem was due to the unaesthetic appearance of the finger extensor spring of the splints. For this reason, modification of these finger extensor outriggers has been attempted. Also, aspects of the splints such as the problems faced and regeneration of the nerve, have been considered and modifications made in order to eliminate any disfunction. This study reflects the reactions and satisfaction of 83 patients concerning the splints they have used. Conventional wire springs have been used in the splints of 71 patients, and 12 had modifications done on their outriggers. In terms of appearance, there were significant differences between modified outriggers and conventional outrigger (p < 0.05). Also, when the modified outriggers were compared in terms of appearance and durability, there was a statistically significant opinion in favour of them (p < 0.01).

Amer, R. K., C. R. Pace-Asciak, et al. (2003). "A lipoxygenase product, hepoxilin A(3), enhances nerve growth factor-dependent neurite regeneration post-axotomy in rat superior cervical ganglion neurons in vitro." Neuroscience 116(4): 935-46.
 Hepoxilins are 12-lipoxygenase metabolites of arachidonic acid found in the CNS. They can modulate neuronal signaling but their functions are not known. We examined the effects of hepoxilin A(3) on neurite outgrowth post-axotomy in an in vitro model of spinal cord transection using superior cervical ganglion neurons. In the absence of nerve growth factor, hepoxilin A(3) did not support neuronal survival, or regeneration post-axotomy but did significantly enhance neurite regeneration in the presence of nerve growth factor. As early as 1 h post-injury hepoxilin A(3)-treated cultures (+nerve growth factor) had significantly more neurites than controls (nerve growth factor alone). Average hourly rates of outgrowth in hepoxilin A(3)-treated cultures were significantly higher than in controls for at least 12 h post-injury, suggesting that the effect of hepoxilin A(3) is maintained in vitro for several hours post-injury. In uninjured neurons hepoxilin A(3) caused a rapid but transient increase in intracellular calcium in the somata; by 2 min post-addition, calcium levels decreased to a new stable plateau significantly higher than pre treatment levels. In injured neurons, hepoxilin A(3) addition immediately post-transection caused a rapid transient increase in intracellular calcium in cell bodies; however, peak calcium levels were significantly lower than in uninjured neurons and the new baseline lower than in uninjured cells. In uninjured cells hepoxilin A(3) addition in zero calcium produced the same pattern, a transient elevation and subsequent decline to a new stable baseline significantly above rest but in injured cells levels fell rapidly to pretreatment values. Taken overall, these findings demonstrate a novel role for hepoxilins as a potentiator of neurite regeneration. They also provide the first evidence that this lipoxygenase metabolite can alter intracellular calcium in neurons by causing release of calcium from intracellular stores and modulating calcium influx mechanisms.

Anderson, K. D., M. A. Merhege, et al. (2003). "Increased expression and localization of the RNA-binding protein HuD and GAP-43 mRNA to cytoplasmic granules in DRG neurons during nerve regeneration." Exp Neurol 183(1): 100-8.
 The neuronal-specific RNA-binding protein, HuD, binds to a U-rich regulatory element of the 3' untranslated region (3' UTR) of the GAP-43 mRNA and delays the onset of its degradation. We have recently shown that overexpression of HuD in embryonic rat cortical cells accelerated the time course of normal neurite outgrowth and resulted in a twofold increase in GAP-43 mRNA levels. Given this evidence, we sought to investigate the involvement of HuD during nerve regeneration. It is known that HuD protein and GAP-43 mRNA are expressed in the dorsal root ganglia (DRG) of adult rat and that GAP-43 is upregulated in DRG neurons during regeneration. In this study, we examined the expression patterns and levels of HuD and GAP-43 mRNA in DRG neurons following sciatic nerve injury using a combination of in situ hybridization, immunocytochemistry, and quantitative RT-PCR. GAP-43 and HuD expression increased in the ipsilateral DRG during the first 3 weeks of regeneration, with peak values seen at 7 days postcrush. At this time point, the levels of HuD and GAP-43 mRNAs in the ipsilateral DRG increased by twofold and sixfold, respectively, relative to the contralateral DRG. Not only were the temporal patterns of expression of HuD protein and GAP-43 mRNA similar, but also they were found to colocalize in the cytoplasm of DRG neurons. Moreover, both molecules were distributed in cytoplasmic granules containing ribosomal RNA. In conclusion, our results suggest that HuD is involved in the upregulation of GAP-43 expression observed at early stages of peripheral nerve regeneration.

Andre, S., H. Boukhaddaoui, et al. (2003). "Axotomy-induced expression of calcium-activated chloride current in subpopulations of mouse dorsal root ganglion neurons." J Neurophysiol 90(6): 3764-73.
 Whole cell patch-clamp recordings of calcium-activated chloride current [ICl(Ca)] were made from adult sensory neurons of naive and axotomized mouse L4-L6 lumbar dorsal root ganglia after 1 day of culture in vitro. A basal ICl(Ca) was specifically expressed in a subset of naive medium-diameter neurons (30-40 microm). Prior nerve injury, induced by sciatic nerve transection 5 days before experiments, increased both ICl(Ca) amplitude and its expression in medium-diameter neurons. Moreover, nerve injury also induced ICl(Ca) expression in a new subpopulation of neurons, the large-diameter neurons (40-50 microm). Small-diameter neurons (inferior to 30 microm) never expressed ICl(Ca). Regulated ICl(Ca) expression was strongly correlated with injury-induced regenerative growth of sensory neurons in vitro and nerve regeneration in vivo. Cell culture on a substrate not permissive for growth, d,l-polyornithine, prevented both elongation growth and ICl(Ca) expression in axotomized neurons. Regenerative growth and the induction of ICl(Ca) expression take place 2 days after injury, peak after 5 days of conditioning in vivo, slowly declining thereafter to control values. The selective expression of ICl(Ca) within medium- and large-diameter neurons conditioned for rapid, efficient growth suggests that these channels play a specific role in postinjury behavior of sensory neuron subpopulations such as neuropathic pain and/or axonal regeneration.

Andre, S., S. Puech-Mallie, et al. (2003). "Axotomy differentially regulates voltage-gated calcium currents in mice sensory neurones." Neuroreport 14(1): 147-50.
 Medium sized dorsal root ganglion neurones are involved in tactile sensation and responsible for allodynia following nerve injury. We examined the effects of sciatic nerve injury on the expression of low and high voltage-gated calcium currents in medium sized neurones isolated from lumbar dorsal root ganglia of adult mice. Based on the relative expression of these calcium channel types, three populations of medium sized neurones were identified in controls. Type I, II and III populations were characterised respectively by small, predominant and no low voltage-gated current compared to the high voltage-gated current. Five days after nerve injury, calcium current expression was differentially affected by axotomy in these three subsets of medium neurones. Altogether, these results suggest that calcium channels are heterogeneously distributed among the medium sized neurones. This heterogeneity should provide specificity not only to sensory functions but also to sensory responses following nerve injury.

Andrews, R. J. (2003). "Neuroprotection trek--the next generation: neuromodulation II. Applications--epilepsy, nerve regeneration, neurotrophins." Ann N Y Acad Sci 993: 14-24; discussion 48-53.
 Three examples of neuroprotective applications of electrical stimulation-neuromodulation-are considered: (1) the diagnosis and treatment of epilepsy, (2) the augmentation of peripheral nerve regeneration after transection, and (3) the interaction between electrical stimulation and neurotrophins (notably brain derived neurotrophic factor [BDNF]) in various neuroprotective situations. The research cited demonstrates clear benefit from appropriate electrical stimulation in the treatment of (1) certain patients with medication-refractory epilepsy, and (2) the functional regeneration of peripheral nerves after transection and surgical repair. Furthermore, neuromodulation of peripheral nerve regeneration has been associated with an increase in the neurotrophin BDNF. The roles of BDNF and other neurotrophins in several disorders of the nervous system are discussed in the context of neuromodulation and its augmentation of neurotrophins. Neuromodulation-at least in part through its effect on BDNF and other neurotrophins-will likely play a major role in the treatment (and possibly prevention) of disorders of the nervous system for which neuroproteive pharmacologic agents have traditionally been sought.

Arlotta, P., S. S. Magavi, et al. (2003). "Induction of adult neurogenesis: molecular manipulation of neural precursors in situ." Ann N Y Acad Sci 991: 229-36.
 Over most of the past century, it was thought that the adult brain was completely incapable of generating new neurons. However, in the last decade, the development of new techniques has resulted in an explosion of new research showing that (i) neurogenesis, the birth of new neurons, is not restricted to embryonic development, but normally also occurs in two limited regions of the adult mammalian brain (the olfactory bulb and the dentate gyrus of the hippocampus); (ii) that there are significant numbers of multipotent neural precursors in many parts of the adult mammalian brain; and (iii) that it is possible to induce neurogenesis even in regions of the adult mammalian brain, like the neocortex, where it does not normally occur, via manipulation of endogenous multipotent precursors in situ. In the neocortex, recruitment of small numbers of new neurons can be induced in a region-specific, layer-specific, and neuronal type-specific manner, and newly recruited neurons can form long-distance connections to appropriate targets. This suggests that elucidation of the relevant molecular controls over adult neurogenesis from endogenous neural precursors/stem cells may allow the development of neuronal replacement therapies for neurodegenerative disease and other central nervous system injuries that may not require transplantation of exogenous cells.

Arlotta, P., S. S. Magavi, et al. (2003). "Molecular manipulation of neural precursors in situ: induction of adult cortical neurogenesis." Exp Gerontol 38(1-2): 173-82.
 Over the past three decades, research exploring potential neuronal replacement therapies have focused on replacing lost neurons by transplanting cells or grafting tissue into diseased regions of the brain. Over most of the past century of modern neuroscience, it was thought that the adult brain was completely incapable of generating new neurons. However, in the last decade, the development of new techniques has resulted in an explosion of new research showing that neurogenesis, the birth of new neurons, normally occurs in two limited and specific regions of the adult mammalian brain, and that there are significant numbers of multipotent neural precursors in many parts of the adult mammalian brain. Recent findings from our lab demonstrate that it is possible to induce neurogenesis de novo in the adult mammalian brain, particularly in the neocortex where it does not normally occur, and that it may become possible to manipulate endogenous multipotent precursors in situ to replace lost or damaged neurons. Recruitment of new neurons can be induced in a region-specific, layer-specific, and neuronal type-specific manner, and newly recruited neurons can form long-distance connections to appropriate targets. Elucidation of the relevant molecular controls may both allow control over transplanted precursor cells and potentially allow the development of neuronal replacement therapies for neurodegenerative disease and other CNS injuries that do not require transplantation of exogenous cells.

Armstrong, J., L. Zhang, et al. (2003). "Axonal regeneration of descending and ascending spinal projection neurons in spinal cord-transected larval lamprey." Exp Neurol 180(2): 156-66.
 The distributions of descending and ascending spinal projection neurons (i.e., spinal neurons with moderate to long axons) were compared in normal larval lamprey and in animals that had recovered for 8 weeks following a complete spinal cord transection at 50% body length (BL, normalized distance from the anterior head). In normal animals, application of HRP to the spinal cord at 60% BL (40% BL) labeled an average of 713.8 +/- 143.2 descending spinal projection neurons (718.4 +/- 108.0 ascending spinal projection neurons) along the rostral (caudal) spinal cord, most of which were unidentified neurons. Some of these neurons project for at least approximately 50-60 spinal cord segments (approximately 36-47 mm in animals with an average length of approximately 90 mm used in the present study). At 8 weeks posttransection, the numbers of HRP-labeled descending or ascending spinal neurons that extended their axons through the transection were about 40% of those in similar areas of the spinal cord in normal animals. Thus, in larval lamprey, axonal regeneration of descending and ascending spinal projection neurons is incomplete, similar to that found for descending brain neurons. The majority of restored projections were from unidentified spinal neurons that have not been documented previously. In contrast to results from several other lower vertebrates, in the lamprey ascending spinal neurons exhibited substantial axonal regeneration. Identified descending spinal neurons, such as lateral interneurons and crossed contralateral interneurons, and identified ascending spinal neurons, such as giant interneurons and edge cells, regenerated their axons at least 9 mm beyond the transection site in animals with an average length of approximately 90 mm, which is appreciably farther than previously reported. In contrast, most dorsal cells, which are centrally located sensory neurons, exhibited very little axonal regeneration.

Au, E. and A. J. Roskams (2003). "Olfactory ensheathing cells of the lamina propria in vivo and in vitro." Glia 41(3): 224-36.
 Olfactory ensheathing cells (OECs) continuously support the regeneration of olfactory receptor neurons (ORNs). In addition, OECs promote regeneration of neurons within the CNS in a number of transplantation paradigms, but details of exactly how they support regeneration remain elusive. The majority of studies using OECs to promote regeneration have thus far focused on understanding the cell biology of OECs purified from the olfactory bulb (OB). Here we show that a population of OECs similar to those obtained from the OB is present in the lamina propria (LP) beneath the olfactory epithelium (OE). These OECs are the first glial cells encountered by the axons of developing ORNs as they exit the OE and display distinct and variable expression of p75, S100beta, GFAP, and O4, characteristic markers of bulb OECs. Once purified in vitro, they display Schwann cell-like and astrocyte-like properties and expand rapidly. In addition to resembling OB-OECs, LP-OECs also express a unique combination of developmentally important proteins-CD 44, beta1 integrin, P200, Notch 3, NG2, VEGF, and PACAP and CREB binding protein (CBP/p300)-not previously reported in OB-OECs. These data suggest that LP-OECs, like OB-OECs, are a developmentally distinct class of glia that are capable of both immature and mature function, depending on environmental stimuli, within the adult nervous system.

Auld, D. S. and R. Robitaille (2003). "Perisynaptic Schwann cells at the neuromuscular junction: nerve- and activity-dependent contributions to synaptic efficacy, plasticity, and reinnervation." Neuroscientist 9(2): 144-57.
 Glial cells are increasingly recognized for their important contributions to CNS and PNS synaptic function. Perisynaptic Schwann cells, which are glial cells at the neuromuscular junction, have proven to be an exceptionally useful model for studying these roles. Recent studies have shown that they detect and reciprocally modulate synaptic efficacy in an activity-dependent manner in the short term. In addition, perisynaptic Schwann cells guide reinnervating nerve sprouts after deinnervation, and many important parameters of this are dependent on synapse activity. Thus, it is hypothesized that perisynaptic Schwann cells are key integrators in a continuum of synaptic efficacy, stability, and plasticity at the neuromuscular junction, which is important for maintaining and restoring synaptic efficacy.

Avramut, M. and C. L. Achim (2003). "Immunophilins in nervous system degeneration and regeneration." Curr Top Med Chem 3(12): 1376-82.
 Immunophilins are receptors for immunosuppressive drugs like cyclosporin A, FK506, rapamycin and their non- immunosuppressive analogs, which are collectively referred to as "immunophilin ligands" (IPL). Cyclosporin A binds to a class of IP called cyclophilins, whereas the receptors for FK506 and rapamycin belong to the family of FK506- binding proteins (FKBP). The latter are designated according to their molecular weight: FKBP12, 25, 52 etc. FKBP levels in the rat brain are up to 50 times higher than in the immune system. FKBP12 is associated with IP3 and ryanodine receptors present on the endoplasmic reticulum and plays a role in stabilizing calcium release. It has also been proposed to be a modulator of the TGFbeta receptor activity. Crush injury of facial or sciatic nerves in rat leads to markedly increased FKBP12 levels in the respective nerve nuclei and this increase is related to nerve regeneration. Cyclophilin A protects cells from death following expression of mutant Cu/ Zn superoxide dismutase, which is associated with familial amyotrophic lateral sclerosis. Our recent studies show that FKBP12 and FKBP52 are expressed in the human nervous system, especially in the substantia nigra- deep gray matter axis. In neurodegenerative diseases, FKBP12 levels increase in neurons situated in areas of pathology. This IP colocalizes with synaptophysin and alpha- synuclein, suggesting that it may become a novel marker of pathology. Immunophilins participate in axonal transport, synaptic vesicle assembly and may play a role in neuroprotection against abnormal protein aggregation, suggesting a potential avenue of therapeutic interventions.

Ayhan, S., N. Markal, et al. (2003). "Effect of subepineurial dehydroepiandrosterone treatment on healing of transected nerves repaired with the epineurial sleeve technique." Microsurgery 23(1): 49-55.
 The epineurial sleeve technique for nerve repair is designed in part to protect a healing nerve from external humoral influences, but research suggests that the external factor dehydroepiandrosterone (DHEA) may actually improve nerve healing in crush injuries. To test the effect of DHEA, we injected it into the epineurial chambers created to repair transected rat sciatic nerves. In 18 control rats, the nerve was transected and repaired without DHEA treatment. Eighteen animals received subepineurial injections of propylene glycol vehicle, and 18 received subepineurial injections of about 0.2 ml DHEA. Walking-track analysis and toe-contracture measurements showed no significant differences among the three groups. At 12 weeks, the gastrocnemius muscles in the DHEA group were significantly heavier than those of untreated controls. At 6 and 12 weeks, DHEA-treated nerves had significantly more myelinated axons, larger average fiber diameter, and greater axonal cross-sectional areas in the proximal, middle, and distal sections. Myelin thickness did not differ between groups, except at 6 weeks between the DHEA and vehicle-treated groups. We conclude that subepineurial dehydroepiandrosterone treatment reduced the extent of denervation atrophy and induced an earlier onset of axonal regeneration.

Bacakoglu, A., M. H. Ozkan, et al. (2003). "Forearm arterial vein grafting: problems and alternative solutions." J Int Med Res 31(5): 458-65.
 We aimed to evaluate patency rates following forearm arterial reconstruction and suggest improvements. Thirty-two vein grafted reconstructions (using saphenous and dorsal hand veins) were evaluated for patency and development of symptoms using clinical examination, Colour-Doppler Sonography and angiography. Overall patency was 59%, with no significant difference between saphenous and dorsal hand veins. Stasis, turbulence, decrease in blood velocity, change in flow pattern, stenosis in the anastomotic area or increase in compliance was detected in 10 patent grafts, but was not associated with symptoms. Six out of 13 non-patent grafted patients had severe or troublesome symptoms associated with accompanying nerve regeneration. To improve patency, careful microsurgical techniques, 'fit vein' and valveless grafts should be used. Dorsal hand veins are most appropriate for short defects. Patency should be evaluated soon after reconstruction.

Bach-y-Rita, P. (2003). "Theoretical basis for brain plasticity after a TBI." Brain Inj 17(8): 643-51.
 Evidence has been accumulating that the brain can reorganize extensively after damage and that reorganization can be obtained even many years after the trauma with appropriate late rehabilitation. An understanding of the brain plasticity mechanisms should lead to more effective rehabilitation and neuropharmacology. In this communication, several emerging concepts with supporting experimental evidence have been presented. These include non-synaptic diffusion neurotransmission, extracellular space volume fraction, neurotransmitters, regeneration and neurogenesis and multiplexing.

Bach, H., D. A. Feldheim, et al. (2003). "Persistence of graded EphA/Ephrin-A expression in the adult frog visual system." J Comp Neurol 467(4): 549-65.
 Many studies have demonstrated the involvement of the EphA family of receptor tyrosine kinases and their ligands, ephrin-A2 and -A5, in the development of the temporonasal axis of the retinotectal/collicular map, but the role of these molecules in optic nerve regeneration has not been well studied. Noting that the characteristic gradients of the EphA/ephrin-A family that are expressed topographically in the retina and tectum of embryonic chicks and mice tend to disappear after birth, we took as our starting point an analysis of EphA and ephrin-A expression in leopard frogs (Rana pipiens and utricularia), species capable of regenerating the retinotectal map as adults. For the EphA family to be involved in the regeneration, one would expect these topographic gradients to persist in the adult or, if downregulated after metamorphosis, to be reexpressed after optic nerve injury. Using EphA3 receptor and ephrin-A5 ligand alkaline phosphatase in situ affinity probes (RAP and LAP, respectively) in whole-mount applications, we report that reciprocally complementary gradients of RAP and LAP binding persist in the optic tract and optic tectum of postmetamorphic frogs, including mature adults. EphA expression in temporal retinal axons in the optic tract was significantly reduced after nerve section but returned during regeneration. However, ephrin-A expression in the tectal parenchyma was not significantly elevated by either eye removal, with degeneration of optic axons, or during regeneration of the retinotectal projection. Thus, the present study has demonstrated a persisting expression of EphA/ephrin-A family members in the retinal axons and tectal parenchyma that may help guide regenerating fibers, but we can offer no evidence for an upregulation of ephrin-A expression in conjunction with optic nerve injury.

Balkowiec-Iskra, E., I. Kurkowska-Jastrzebska, et al. (2003). "Post intoxicative therapeutic immunization with myelin oligodendrocyte glycoproteine (MOG 35-55) suppresses spontaneous regeneration of dopaminergic neurons injured with 1-methyl-4 phenyl-1,2,3,6-tetrahydropiridine (MPTP)." Acta Neurobiol Exp (Wars) 63(2): 109-15.
 The pathological process of neurodegeneration is accompanied by an inflammatory reaction that is believed to contribute to the pathogenesis of neurodegenerative diseases. The aim of our study was to evaluate the influence of autoimmune reaction induced by post-traumatic vaccination with myelin self-antigen on spontaneous regeneration of dopaminergic neurons, injured with MPTP. C57BL mice were intoxicated with 40 mg/kg MPTP and seven days later immunized with MOG 35-55 peptide in CFA. On the 7th day following intoxication, the MPTP treated mice showed decrease of the dopamine level by 63% as compared to the control mice. However, starting from the 14th day following intoxication, a spectacular increase of dopamine content was observed. Immunization with MOG resulted in a statistically significant reduction of the increase in striatum as compared to non-immunized animals, and was lower by 23%, 17% and 15% on days 14, 28 and 50, respectively. Our results show suppressive influence of autoimmune reaction induced after injury on regeneration of dopamine cells intoxicated with MPTP.

Bambakidis, N. C., R. Z. Wang, et al. (2003). "Sonic hedgehog-induced neural precursor proliferation after adult rodent spinal cord injury." J Neurosurg 99(1 Suppl): 70-5.
 OBJECT: The glycoprotein molecule sonic hedgehog (Shh) has been shown to play a critical role in neuraxial development. To assess its role in the repair of demyelination following spinal cord injury (SCI), escalating doses of Shh were injected into demyelinated lesions in adult rat spinal cords. METHODS: Twenty-seven adult rats underwent thoracic laminectomy and chemical demyelination of the spinal cord dorsal columns without neurological deficit A subset of 20 rats was treated after 3 days by direct injection of Shh at two different doses. Rats were killed at 7 or 21 days after SCI, and tissue samples underwent immediate fixation or were placed into cell culture. Diffuse cellular proliferative responses throughout the gray and white matter were observed in up to 70% of Shh-treated rats. Proliferation around the central canal, believed to be derived from the ventricular ependyma consistent with neuronal stem cell induction, was demonstrated in up to 60% of the treated rats. No significant proliferation in these areas was detected in control rats. Dorsal areas of nestin-positive cells were also observed in 70% of rats treated with high doses of Shh, and these observations were reproduced in cell culture as well as in cultures of dorsal spinal cord explants. Cell counts revealed significant increases in the percentage of oligodendrocyte precursors and neurons in treated compared with control rats. CONCLUSIONS: Exogenous Shh administration promotes nestin-positive cell proliferation after SCI in adult rodents. These cells are believed to be derived from neural precursor cells. The populations of oligodendrocyte precursors and neurons were likewise increased in Shh-treated rats, suggesting that these cells may be derived from neural stem cells.

Bandtlow, C. E. (2003). "Regeneration in the central nervous system." Exp Gerontol 38(1-2): 79-86.
 Unlike neonatal axons, mammalian adult axons of the CNS do not regenerate after injury. This developmental loss of regenerative capacity, is correlated with the onset of myelination. Likewise, myelin, or myelin-associated components such as Nogo-A and myelin-associated glycoprotein (MAG) inhibit regeneration from older but not younger neurons. Identification of the molecular events responsible for this developmental loss of regenerative capacity is central to devise strategies to encourage regeneration in adults after injury. Endogenous levels of the cyclic nucleotides cAMP and cGMP have been suggested to determine the neuronal responsiveness to various axonal guidance factors. Elevating cAMP concentrations block Nogo-A or MAG induced inhibition of neurite outgrowth in older neurons, whereas suppressing cAMP levels in young neurons renders them susceptible to Nogo-A and MAG. Interestingly, elevated cAMP levels abrogated the Nogo-A and MAG mediated activation of RhoA and down regulation of Rac1 in adult neurons. In contrast, elevation of cAMP leads to the inactivation of RhoA and prevents activation of downstream effector proteins, while Rac is activated. We therefore conclude that the endogenous neuronal cAMP levels determine the neuronal responsiveness to myelin-associated neurite growth inhibitors by regulating rho GTPase activities.

Barcelos, A. S., A. C. Rodrigues, et al. (2003). "Inside-out vein graft and inside-out artery graft in rat sciatic nerve repair." Microsurgery 23(1): 66-71.
 Although veins and arteries present similar wall structures, there are differences which may be relevant in peripheral nerve reconstruction. Inside-out vein grafts (IOVG) have been satisfactorily used to repair both motor and sensitive nerves. However, the inside-out artery graft (IOAG) is a new technique and not fully investigated. Our study presents comparative morphological data on nerve regeneration achieved with IOVG and IOAG in the repair of Wistar rat sciatic nerves. Jugular veins and aorta arteries were harvested from donor animals and used "inside-out" to bridge a 10-mm gap. Animals were sacrificed at 10 weeks to evaluate nerve regeneration. Both techniques presented great variability in nervous tissue, though some animals showed satisfactory results. Different intensities of scarring processes might have interfered with nerve regeneration. Although IOVG and IOAG techniques showed similar morphometric results, in general, IOVG presented a closer-to-normal nerve organization than IOAG.

Bareyre, F. M. and M. E. Schwab (2003). "Inflammation, degeneration and regeneration in the injured spinal cord: insights from DNA microarrays." Trends Neurosci 26(10): 555-63.
 
Barres, B. A. (2003). "What is a glial cell?" Glia 43(1): 4-5.
 
Beazley, L. D., J. Rodger, et al. (2003). "Training on a visual task improves the outcome of optic nerve regeneration." J Neurotrauma 20(11): 1263-70.
 Optic nerve regeneration in a lizard, Ctenophorus ornatus, is dysfunctional despite survival of most retinal ganglion cells and axon regeneration to the optic tectum. The regenerated retino-tectal projection at 6 months has crude topography but by 1 year is disordered; visually-elicited behavior is absent via the experimental eye. Here, we assess the influence of training on the outcome of optic nerve regeneration. Lizards were trained to catch prey presented within the monocular field of either eye. One optic nerve was then severed and visual stimulation resumed throughout regeneration. In the trained group, presentation was restricted to the eye undergoing optic nerve regeneration; for the untrained group, the unoperated eye was stimulated. Pupil responses returned in trained but not in untrained animals. At 1 year, trained animals oriented to and captured prey; untrained animals demonstrated minimal orienting and failed to capture prey. Regenerated retino-tectal projections were topographic in the trained but not in the untrained group as assessed by in vitro electrophysiological recording and by carbocyanine dye tracing. In vitro electrophysiological recording during application of neurotransmitter antagonists to the tectum revealed that the level of GABAergic inhibition was modest in trained animals but elevated in the untrained group; responses were mainly AMPA-mediated in both groups. We conclude that training improves the behavioral outcome of regeneration, presumably by stabilizing and refining the transient retino-tectal map and preventing a build-up of tectal inhibition. The results suggest that for successful central nerve regeneration to occur in mammals, it may be necessary to introduce training to complement procedures stimulating axon regeneration.

Becker, D., C. L. Sadowsky, et al. (2003). "Restoring function after spinal cord injury." Neurolog 9(1): 1-15.
 BACKGROUND: By affecting young people during the most productive period of their lives, spinal cord injury is a devastating problem for modern society. A decade ago, treating SCI seemed frustrating and hopeless because of the tremendous morbidity and mortality, life-shattering impact, and limited therapeutic options associated with the condition. Today, however, an understanding of the underlying pathophysiological mechanisms, the development of neuroprotective interventions, and progress toward regenerative interventions are increasing hope for functional restoration. REVIEW SUMMARY: This study addresses the present understanding of SCI, including the etiology, pathophysiology, treatment, and scientific advances. The discussion of treatment options includes a critical review of high-dose methylprednisolone and GM-1 ganglioside therapy. The concept that limited rebuilding can provide a disproportionate improvement in quality of life is emphasized throughout. CONCLUSIONS: New surgical procedures, pharmacologic treatments, and functional neuromuscular stimulation methods have evolved over the last decades that can improve functional outcomes after spinal cord injury, but limiting secondary injury remains the primary goal. Tissue replacement strategies, including the use of embryonic stem cells, become an important tool and can restore function in animal models. Controlled clinical trials are now required to confirm these observations. The ultimate goal is to harness the body's own potential to replace lost central nervous system cells by activation of endogenous progenitor cell repair mechanisms.

Bedogni, B., G. Pani, et al. (2003). "Redox regulation of cAMP-responsive element-binding protein and induction of manganous superoxide dismutase in nerve growth factor-dependent cell survival." J Biol Chem 278(19): 16510-9.
 Reactive oxygen species (ROS) act as both signaling molecules and mediators of cell damage in the nervous system and are implicated in the pathogenesis of neurodegenerative diseases. Neurotrophic factors such as the nerve-derived growth factor (NGF) support neuronal survival during development and promote regeneration after neuronal injury through the activation of intracellular signals whose molecular effectors and downstream targets are still largely unknown. Here we present evidence that early oxidative signals initiated by NGF in PC12 cells, an NGF-responsive cell line, play a critical role in preventing apoptosis induced by serum deprivation. This redox-signaling cascade involves phosphatidylinositol 3-kinase, the small GTPase Rac-1, and the transcription factor cAMP-responsive element-binding protein (CREB), a molecule essential to promote NGF-dependent survival. We found that ROS are necessary for NGF-dependent phosphorylation of CREB, an event directly correlated with CREB activity, whereas hydrogen peroxide induces a robust CREB phosphorylation. Cells exposed to NGF show a late decrease in the intracellular content of ROS when compared with untreated cells and increased expression of the mitochondrial antioxidant enzyme manganese superoxide dismutase, a general inhibitor of cell death. Accordingly, serum deprivation-induced apoptosis was selectively inhibited by low concentrations of the mitochondrially targeted antioxidant Mito Q (mitoquinol/mitoquinone). Taken together, these data demonstrate that the oxidant-dependent activation of CREB is a component of NGF survival signaling in PC12 cells and outline an intriguing circuitry by which a cytosolic redox cascade promotes cell survival at least in part by increasing mitochondrial resistance to oxidative stress.

Befort, K., L. Karchewski, et al. (2003). "Selective up-regulation of the growth arrest DNA damage-inducible gene Gadd45 alpha in sensory and motor neurons after peripheral nerve injury." Eur J Neurosci 18(4): 911-22.
 The growth arrest and DNA damage-inducible gene 45 alpha (Gadd45a) was one of 240 genes found previously by high density oligonucleotide microarray analysis to be regulated in the rat L4 and L5 dorsal root ganglia 3 days after transection of the sciatic nerve (>four-fold up-regulation). The Gadd45a mRNA expression profile investigated by northern blot, RNase protection assay and in situ hybridization in the rat shows negligible constitutive mRNA levels in embryonic, neonatal or adult intact dorsal root ganglia. Within 24 h of a sciatic nerve injury, a very large induction is found that persists for as long as regeneration of injured fibres is prevented by peripheral nerve ligation. When axons are allowed to regrow following sciatic nerve crush injury, Gadd45a expression is terminated at later time points, when levels of other markers of injury return towards normal. Colocalization with activating transcription factor 3-LI and c-jun mRNA implies that all peripherally injured primary sensory and motor neurons express Gadd45a mRNA. Injury to the central axons of dorsal root ganglion neurons produces only a minimal induction of Gadd45a while peripheral inflammation is without effect. Gadd45a is a specific marker of the presence of peripheral axonal injury in adult primary sensory and motor neurons.

Belecky-Adams, T., M. Holmes, et al. (2003). "An intact intermediate filament network is required for collateral sprouting of small diameter nerve fibers." J Neurosci 23(28): 9312-9.
 Expression of the intermediate filament (IF) protein peripherin is initiated during development at the time of axonal extension and increases during regeneration of nerve fibers. To test whether the IF network is essential for neuron process outgrowth in the mature organism in vivo, we disrupted endogenous peripherin IF in small-sized dorsal root ganglion (DRG) neurons in transgenic mice via expression of a mutant peripherin transgene under control of peripherin gene regulatory sequences. Anatomical and functional analyses showed that these neurons send peripheral and central axonal projections to correct targets, express correct neuropeptides, and mediate acute pain responses normally. However, disruption of IF significantly impaired the ability of uninjured small-sized DRG neurons to sprout collateral axons into adjacent denervated skin, indicating a critical role for intact IF in plasticity, specifically in compensatory nociceptive nerve sprouting.

ben-Aaron, M. (2003). "Topological aspects of axonal regeneration." Med Hypotheses 61(5-6): 597-600.
 The fibers making up any sensory system in the spinal cord come from the same cells as do the fibers in peripheral nerves yet severed nerve fibers in the adult spinal cord do not regenerate but damaged peripheral nerves - those in the extremities - do heal themselves. Why should spinal cord regeneration even be an issue, why should an inhibiting protein have evolved to prevent it and what causes this protein to be expressed? From a holistic perspective, an answer to this conundrum shows that these questions are intertwined, and suggests that: (1) The model of the neurons as 'wires' is too simplistic. (2) In humans, the 'map' of individual connections is (topologically, at least) locally variable, though the overall global topology and 'functionality' of each normal spinal cord is constant. Both of these issues have to be addressed if functional restoration is to be achieved.

Benitez-Temino, B., R. R. de la Cruz, et al. (2003). "Grafting of a new target prevents synapse loss in abducens internuclear neurons induced by axotomy." Neuroscience 118(3): 611-26.
 The loss of afferent synaptic boutons is a prominent alteration induced by axotomy on adult central neurons. In this work we attempted to prove whether synapse loss could be reverted by reconnection with a new target. We severed the medial longitudinal fascicle of adult cats and then transplanted embryonic cerebellar primordia at the lesion site immediately after lesion. As previously shown, the transected axons from abducens internuclear neurons penetrate and reinnervate the graft [J Comp Neurol 444 (2002) 324]. By immunocytochemistry and electron microscopy we studied the synaptology of abducens internuclear neurons under three conditions: control, axotomy and transplant (2 months of survival time). Semithin sections of the abducens nucleus were immunostained against calretinin, to identify abducens internuclear neurons, and either synaptophysin (SF), to label synaptic terminals, or glial fibrillary acidic protein (GFAP) to detect the astrocytic reaction. Optical and linear density of SF and GFAP immunostaining were measured. Data revealed a significant decrease in the density of SF-labeled terminals with a parallel increase in GFAP-immunoreactive elements after axotomy. On the contrary, in the transplant group, the density of SF-labeled terminals was found similar to control, and the astrocytic reaction induced by lesion was significantly reduced. At the ultrastructural level, synaptic coverage and linear density of boutons were measured around the somata of abducens internuclear neurons. Whereas a significant reduction in both parameters was found after axotomy, cells of the transplant group received a normal density of synaptic endings. The ratio between F- and S-type boutons was found similar in the three groups. Therefore, these findings indicate that the grafting of a new target can prevent the loss of afferent synaptic boutons produced by the axotomy.

Benjamin, B. (2003). "Vocal cord paralysis, synkinesis and vocal fold motion impairment." ANZ J Surg 73(10): 784-6.
 Felix Semon's 'laws' of vocal cord paralysis were conceived over a century ago, based on the simple concept that abductor function of the recurrent laryngeal nerve was more vulnerable than adductor function. It is now clear that the neuromuscular pathology of laryngeal innervation is much more complex. Whether the nerve has been cut, crushed, stretched, cauterized or otherwise injured, it is seldom completely transected. There might be no detectable vocal cord movement at laryngoscopy, yet, electromyography usually shows at least some activity because of incomplete denervation and/or developing synkinesis. Electrical silence hardly ever persists forever. Disordered vocal fold movement following nerve injury appears to depend on laryngeal synkinesis with disorganized neuromuscular function caused by misdirected regeneration and aberrant reinnervation, sometimes by adjacent nerves. The severity of the injury, abnormal random reinnervation, scar tissue formation and nerve growth-stimulating and inhibiting factors influence the final position of the vocal fold. For a better understanding of neurolaryngological disorders it is no longer sufficient to think merely in terms of 'vocal cord paralysis'.

Bermingham-McDonogh, O. and E. W. Rubel (2003). "Hair cell regeneration: winging our way towards a sound future." Curr Opin Neurobiol 13(1): 119-26.
 The discovery of hair cell regeneration in the inner ear of birds provides new optimism that there may be a treatment for hearing and balance disorders. In this review we describe the process of hair cell regeneration in birds; including restoration of function, recovery of perception and what is currently known about molecular events, such as growth factors and signalling systems. We examine some of the key recent findings in both birds and mammals.

Bertelli, J. A. and M. F. Ghizoni (2003). "Nerve repair by end-to-side coaptation or fascicular transfer: a clinical study." J Reconstr Microsurg 19(5): 313-8.
 End-to-side nerve repair has recently achieved special interest in the laboratory. In clinical practice, its use remains controversial and very few studies have been published. In the present report, nerve repair by end-to-side neurorrhaphy and by fascicular transfer is evaluated in seven patients. Clinical, electrophysiologic and histologic recovery was not observed in the patients who underwent end-to-side nerve coaptation, but recovery was observed when a fascicular transfer was performed.

Bertelli, J. A. and M. F. Ghizoni (2003). "Brachial plexus avulsion injury repairs with nerve transfers and nerve grafts directly implanted into the spinal cord yield partial recovery of shoulder and elbow movements." Neurosurgery 52(6): 1385-9; discussion 1389-90.
 OBJECTIVE: Complete avulsion of the brachial plexus is a devastating injury that primarily affects young adults. The current treatment is based on nerve transfers, which yield very limited recovery. In this study, brachial plexus injuries were repaired with nerve transfers and nerve grafts directly implanted into the spinal cord. METHODS: Eight patients with complete brachial plexus avulsion injuries were surgically treated. Roots or target nerves of the brachial plexus were repaired with peripheral nerve grafts directly implanted into the spinal cord and with extraplexal nerve transfers. RESULTS: Muscle reinnervation was observed for six patients who received spinal implants. Among those patients, one recovered M4 muscle power. Reinnervation was observed only in proximal upper limb muscles. CONCLUSION: Muscle reinnervation through nerve grafts directly implanted into the spinal cord was demonstrated. It seems that the combination of intra- and extradural neurotizations improves the proximal muscle function results. However, the extent of this improvement is limited and, in our opinion, does not justify the use of spinal implants.

Biers, S. M. and A. F. Brading (2003). "Nerve regeneration: might this be the only solution for functional problems of the urinary tract?" Curr Opin Urol 13(6): 495-500.
 SUMMARY: PURPOSE OF REVIEW To assess the potential role of nerve regeneration in restoring urinary tract function, the rapidly developing and exciting area of central and peripheral nerve repair and regeneration is reviewed, with particular reference to papers in which animal models of nerve damage resulting in urogenital dysfunction have been used. The difficulties and potential of these techniques for therapeutic application to human subjects with functional problems of the urinary tract are discussed.RECENT FINDINGS Methods for encouraging regeneration of cut axons and directed growth in the inhibitory environment of the central nervous system are being extensively explored. The recent discovery of the potential of olfactory ensheathing cells has proved a significant advance. Olfactory ensheathing cells are a type of glial cell which can be harvested from the olfactory mucosa. Transplantation of these cells, in conjunction with a biodegradable synthetic nerve guide or conduit, has been shown to restore urinary tract function after spinal cord injury. Artificial, biodegradable conduits have also restored bladder and spermatic duct function after sympathetic nerve damage. Other adjuvants facilitating the process of axonal recovery include the use of neurotrophins to accelerate and guide the formation of new nerve-fibre growth.SUMMARY These revolutionary technologies may, in the future, provide a means of treating urinary tract dysfunction with some types of aetiology, including acute spinal cord injury, and injury to nerves following pelvic surgery. It is, however, less likely that these treatments will be used successfully in the near future in patients in which the neural damage is long term, or associated with death of post-ganglionic neurons.

Biran, R., M. D. Noble, et al. (2003). "Directed nerve outgrowth is enhanced by engineered glial substrates." Exp Neurol 184(1): 141-52.
 In the present study, the influence of astrocyte alignment on the direction and length of regenerating neurites was examined in vitro. Astrocytes were experimentally manipulated by different approaches to create longitudinally aligned monolayers. When cultured on the aligned monolayers, dorsal root ganglion neurites grew parallel to the long axis of the aligned astrocytes and were significantly longer than controls. Engineered monolayers expressed linear arrays of fibronectin, laminin, neural cell adhesion molecule, and chondroitin sulfate proteoglycan that were organized parallel to one another, suggesting that a particular spatial arrangement of these molecules on the astrocyte surface may be necessary to direct nerve regeneration in vivo. In contrast, no bias in directional outgrowth was observed for neurites growing on unorganized monolayers. The results suggest that altering the organization of astrocytes and their scar-associated matrix at the lesion site may be used to influence the direction and the length of adjacent regenerating axons in the damaged brain and spinal cord.

Bird, E. V., F. M. Boissonade, et al. (2003). "Neuropeptide expression following ligation of the ferret lingual nerve." Arch Oral Biol 48(7): 541-6.
 Previous studies on the ferret inferior alveolar nerve found a close association between the spontaneous neural activity generated at a site of nerve injury, and the accumulation of neuropeptides in the injured axons. More recent electrophysiological studies on the lingual nerve revealed high levels of spontaneous activity 3 days after injury, a decline at 3 weeks and a late rise at 3 months. In the present study we have used immunocytochemical techniques to see whether this time course of spontaneous activity is again paralleled by an accumulation of neuropeptides. In 20 anaesthetised adult ferrets the left lingual nerve was ligated and sectioned distally, and the animals left to recover for 3 days, 3 weeks or 3 months. The tissue was processed using indirect immunofluorescence and image analysis was used to quantify levels of the neuropeptides; calcitonin gene-related peptide (CGRP), substance P (SP), vasoactive intestinal polypeptide (VIP), galanin (GAL), enkephalin (ENK) and neuropeptide Y (NPY). Immunoreactivity to all of the neuropeptides was present proximal to the ligature 3 days after injury, and these high levels of expression had decreased considerably by 3 weeks. By 3 months ENK and NPY expression had almost disappeared proximal to the ligature, but levels of CGRP, SP, VIP and GAL had increased slightly. This was also accompanied by an accumulation of all of the neuropeptides, except NPY, in the portion of nerve immediately distal to the ligature. This late accumulation of certain neuropeptides coincides with the increase in spontaneous activity seen in our previous electrophysiological studies and supports the suggestion that neuropeptides may play a role in the aetiology of sensory disorders after nerve injury.

Birouk, N., H. Azzedine, et al. (2003). "Phenotypical features of a Moroccan family with autosomal recessive Charcot-Marie-Tooth disease associated with the S194X mutation in the GDAP1 gene." Arch Neurol 60(4): 598-604.
 BACKGROUND: The first locus for demyelinating autosomal recessive Charcot-Marie-Tooth (ARCMT) disease was identified in 8q13, where mutations in GDAP1 have been found. Mutations in the same gene have been detected in families with axonal ARCMT disease. OBJECTIVE: To determine the clinical, electrophysiologic, and morphologic characteristics of a consanguineous Moroccan family with ARCMT disease associated with the S194X mutation in the GDAP1 gene. METHODS: Four patients from a consanguineous Moroccan family were examined clinically and electrophysiologically. In one patient, a morphometric and ultrastructural study of a peroneal nerve biopsy sample was performed. Mutation in the coding region of the GDAP1 gene was identified by direct sequencing. RESULTS: Neuropathy was evident early in childhood, walking was delayed in one patient, and onset of symptoms occurred before 18 months in the others. The phenotype was severe: foot deformities and disabilities involving the hands and feet developed toward the end of the first decade, followed by involvement of proximal muscles in the lower limbs, leading to loss of autonomy. Electrophysiologic findings were consistent with an axonal form of CMT disease: motor nerve conduction velocities, recordable in one patient only, were greater than 40 m/sec. Sensory nerve action potentials were either abolished or substantially reduced in amplitude. The morphologic data supported the diagnosis of axonal neuropathy, showing a marked reduction in myelinated fibers and signs of axonal regeneration, including frequent pseudo-onion bulb formations. The 4 patients in this family were homozygous for the S194X mutation in the GDAP1 gene. CONCLUSION: Electrophysiologic and pathological findings support the hypothesis of an axonal disorder in this ARCMT family with the S194X mutation in the GDAP1 gene.

Blaauw, G. and A. C. Slooff (2003). "Transfer of pectoral nerves to the musculocutaneous nerve in obstetric upper brachial plexus palsy." Neurosurgery 53(2): 338-41; discussion 341-2.
 OBJECTIVE: To investigate the results of transfer of pectoral nerves to the musculocutaneous nerve for treatment of obstetric brachial palsy. METHODS: In 25 cases of obstetric brachial palsy (20 after breech deliveries), branches of the pectoral nerve plexus were transferred directly to the musculocutaneous nerve. For all patients, the nerve transfer was part of an extended brachial plexus reconstruction. Results were tested both clinically and with the Mallet scale, at a mean follow-up time of 70 months (standard deviation, 34.3 mo). RESULTS: There were two complete failures, which were attributable to disconnection of the transferred nerve endings. The results after transfer were excellent in 17 cases and fair in 5 cases. Steindler flexorplasty improved elbow flexion for three patients. CONCLUSION: Transfer of pectoral nerves to the musculocutaneous nerve for treatment of obstetric upper brachial palsy may be effective, if the specific anatomic features of the pectoral nerve plexus are sufficiently appreciated.

Blits, B., M. Oudega, et al. (2003). "Adeno-associated viral vector-mediated neurotrophin gene transfer in the injured adult rat spinal cord improves hind-limb function." Neuroscience 118(1): 271-81.
 To foster axonal growth from a Schwann cell bridge into the caudal spinal cord, spinal cells caudal to the implant were transduced with adeno-associated viral (AAV) vectors encoding for brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (AAV-NT-3). Control rats received AAV vectors encoding for green fluorescent protein or saline. AAV-BDNF- and AAV-NT-3-transduced 293 human kidney cells produced and secreted BDNF or NT-3, respectively, in vitro. The secreted neurotrophins were biologically active; they both promoted outgrowth of sensory neurites in vitro. In vivo, transgene expression was observed predominantly in neurons for at least 16 weeks after injection. Compared with controls, a modest though significant improvement in hind-limb function was found in rats that received AAV-BDNF and AAV-NT-3. Retrograde tracing demonstrated that twice as many neurons with processes extending toward the Schwann cell graft were present in the second lumbar cord segment of AAV-BDNF- and AAV-NT-3-injected animals compared with controls. We found no evidence, however, for growth of regenerated axons from the Schwann cell implant into the caudal cord.Our results suggest that AAV vector-mediated overexpression of BDNF and NT-3 in the cord caudal to a Schwann cell bridge modified the local lumbar axonal circuitry, which was beneficial for locomotor function.

Bontioti, E. N., M. Kanje, et al. (2003). "Regeneration and functional recovery in the upper extremity of rats after various types of nerve injuries." J Peripher Nerv Syst 8(3): 159-68.
 The aim was to establish an accurate, reproducible, and simple method to evaluate functional recovery after different types of nerve injuries to the brachial plexus of rats. To that end, pawprints, measured as distance between the first and fourth and second and third digits, were used for evaluation of injuries including crush injury, transection/repair, or graft repair of the median, ulnar, and radial nerves. Immunocytochemistry of the C-terminal flanking peptide of neuropeptide Y (CPON) and neurofilaments was used to investigate the cell body response and axonal outgrowth, respectively. Functional recovery was dependent on the severity as well as on the level of the lesion. Neither a single injury to the median nerve nor an injury to the ulnar nerve affected the pawprint, while an injury to both these nerves or a single injury to the radial nerve caused impairment of pawprints. There was a rapid recovery after crush injury to these nerves compared to previous reports of a similar injury to the sciatic nerve. The pattern of axonal outgrowth was related to the severity of the lesion. A conditioning lesion, i.e., an initial lesion of the same nerve preceding a test injury by a few days, of both motor/sensory fibers led to a quicker functional recovery. Surprisingly, conditioning of only sensory fibers had nearly the same effect. The cell body response was dependent on the level of the nerve lesion. The upper extremity of rats might be useful to evaluate the effects of new repair methods after nerve injuries using functional evaluation with pawprints as a simple and accurate method.

Borgens, R. B. (2003). "Restoring function to the injured human spinal cord." Adv Anat Embryol Cell Biol 171: III-IV, 1-155.
 Of catastrophic traumas to the human body, spinal cord injury (SCI) has least benefited innovations arising from the new biology. Since after WW II, the "standard of care" for SCI has changed little. The controversial use of high dosages of steroids has provided only modest benefit to patients--but not without the enhanced risk of mortality. Novel therapies arising from biochemistry and genetics have not materialized in over 15 years, and are unlikely to in the author's opinion. Instead, appreciation of biophysics and cell physiology in controlling nerve injury, growth, regeneration, and function has produced innovative clinical approaches now in testing in human spinal cord injury.

Borisoff, J. F., C. C. Chan, et al. (2003). "Suppression of Rho-kinase activity promotes axonal growth on inhibitory CNS substrates." Mol Cell Neurosci 22(3): 405-16.
 Several molecules inhibit axonal growth cones and may account for the failure of central nervous system regeneration, including myelin proteins and various chondroitan sulfate proteoglycans expressed at the site of injury. Axonal growth inhibition by myelin and chondroitan sulfate proteoglycans may in part be controlled by Rho-GTPase, which mediates growth cone collapse. Here, we tested in vitro whether pharmacological inhibition of a major downstream effector of Rho, Rho-kinase, promotes axonal outgrowth from dorsal root ganglia grown on aggrecan. Aggrecan substrates stimulated Rho activity and were inhibitory to axonal growth. Y-27632 treatment promoted the growth of axons by 5- to 10-fold and induced "steamlined" growth cones with longer filopodia and smaller lamellipodia. Interestingly, more actin bundles reminiscent of stress fibers in the central domain of the growth cone were observed when grown on aggrecan compared to laminin. In addition, Y-27632 significantly promoted axonal growth on both myelin and adult rat spinal cord cryosections. Our data suggest that suppression of Rho-kinase activity may enhance axonal regeneration in the central nervous system.

Borschel, G. H., K. F. Kia, et al. (2003). "Mechanical properties of acellular peripheral nerve." J Surg Res 114(2): 133-9.
 BACKGROUND: Acellular nerve has been used in experimental models as a peripheral nerve substitute. Our objective was to determine the difference in tensile strength between fresh and chemically treated acellularized peripheral nerve. MATERIALS AND METHODS: F344 rat sciatic nerves were either fresh or acellularized and tested either whole (Part A) or transected and repaired (Part B). For all constructs, the mean ultimate stress, mean ultimate strain, Young's modulus, and total mechanical work to fracture were calculated.The average ultimate strains for Groups A-1 and A-2 were 0.480 +/- 0.117 and 0.810 +/- 0.114, respectively. The Young's moduli in Groups A-1 and A-2 were 576 +/- 160 and 580 +/- 150 kPa, respectively. In Groups A-1 and A-2, the normalized work to failure was 0.35 +/- 0.14 and 1.11 +/- 0.38 N. The specimens in Group B-1 withstood an average ultimate stress of 780 +/- 280 kPa. The specimens in Group B-2 withstood an average ultimate stress of 405 +/- 20 kPa. RESULTS: The average ultimate strains for Groups B-1 and B-2 were 0.319 +/- 0.087 and 0.266 +/- 0.019, respectively. The Young's moduli in Groups B-1 and B-2 were 4,030 +/- 1360 and 2,290 +/- 280 kPa, respectively. The normalized work to failure in Groups B-1 and B-2 was calculated as 0.22 +/- 0.04 and 0.11 +/- 0.02 N. CONCLUSIONS: Although adequately robust for reconstructive procedures, the acellular peripheral nerve had decreased tensile strength compared with fresh nerve either when tested whole or when transected and repaired.

Bouslama-Oueghlani, L., R. Wehrle, et al. (2003). "The developmental loss of the ability of Purkinje cells to regenerate their axons occurs in the absence of myelin: an in vitro model to prevent myelination." J Neurosci 23(23): 8318-29.
 Axonal regeneration in the mammalian CNS is a property of immature neurons that is lost during development. Using organotypic culture of cerebellum, we have shown that in vitro Purkinje cells lose their regenerative capacity in parallel with the process of myelination. We have investigated whether myelination is involved in the age-dependent loss of regeneration of these neurons. By applying a high dose of bromodeoxyuridine in the culture medium of newborn cerebellar slices during the first 3 d in vitro, we have succeeded in obtaining cultures with oligodendrocyte depletion, together with a lack of ameboid microglia and enhancement of Purkinje cell survival. These cultures, after 14 d in vitro, are completely devoid of myelin. We have compared the ability of Purkinje cells to regenerate their axons in the presence or absence of myelin. Purkinje cells in cerebellar explants taken at birth, treated with bromodeoxyuridine and axotomized after 7 d in vitro, survive better than similar neurons in untreated cultures. However, despite the lack of myelin and the enhanced survival, Purkinje cells do not regenerate, whereas they do regenerate when the axotomy is done at postnatal day 0. Thus, the Purkinje cell developmental switch from axonal regeneration to lack of regeneration does not appear to be regulated by myelin.

Boyd, J. G. and T. Gordon (2003). "Glial cell line-derived neurotrophic factor and brain-derived neurotrophic factor sustain the axonal regeneration of chronically axotomized motoneurons in vivo." Exp Neurol 183(2): 610-9.
 In contrast to injuries in the central nervous system, injured peripheral neurons will regenerate their axons. However, axotomized motoneurons progressively lose their ability to regenerate their axons, following peripheral nerve injury often resulting in very poor recovery of motor function. A decline in neurotrophic support may be partially responsible for this effect. The initial upregulation of glial cell line-derived neurotrophic factor (GDNF) and brain-derived neurotrophic factor (BDNF) by Schwann cells of the distal nerve stump after nerve injury has led to the speculation that they are important for motor axonal regeneration. However, few experiments directly measure the effects of exogenous BDNF or GDNF on motor axonal regeneration. This study provided the first direct and quantitative evidence that long-term continuous treatment with exogenous GDNF significantly increased the number of motoneurons which regenerate their axons, completely reversing the negative effects of chronic axotomy. The beneficial effect of GDNF was not dose-dependent. A combination of exogenous GDNF and BDNF on motor axonal regeneration was significantly greater than either factor alone, and this effect was most pronounced following long-term continuous treatment. The ability of GDNF, either alone or in combination with BDNF, to increase the number of motoneurons that regenerated their axons correlated well with an increase in axon sprouting within the distal nerve stump. Thus long-term continuous treatment with neurotrophic factors, such as GDNF and BDNF, can be used as a viable treatment to sustain motor axon regeneration.

Boyd, J. G., V. Skihar, et al. (2003). "Olfactory ensheathing cells: historical perspective and therapeutic potential." Anat Rec 271B(1): 49-60.
 Olfactory ensheathing cells (OECs) are the glial cells that ensheath the axons of the first cranial nerve. They are attracting increasing attention from neuroscientists as potential therapeutic agents for use in the repair of spinal cord injury and as a source of myelinating glia for use in remyelinating axons in demyelinating diseases such as multiple sclerosis. This review mainly addresses the cell biological aspects of OECs pertinent to addressing two questions. Namely, where do OECs fit into the groupings of central nervous system (CNS)/peripheral nervous system (PNS) glial cells and should OECs be viewed as a clinically relevant alternative to Schwann cells in the treatment of spinal cord injury? The evidence indicates that OECs are indeed a clinically relevant alternative to Schwann cells. However, much more work needs to be done before we can even come close to answering the first question as to the lineage and functional relationship of OECs to the other types of CNS and PNS glial cells.

Boyd, J. G. and T. Gordon (2003). "Neurotrophic factors and their receptors in axonal regeneration and functional recovery after peripheral nerve injury." Mol Neurobiol 27(3): 277-324.
 Over a half a century of research has confirmed that neurotrophic factors promote the survival and process outgrowth of isolated neurons in vitro. The mechanisms by which neurotrophic factors mediate these survival-promoting effects have also been well characterized. In vivo, peripheral neurons are critically dependent on limited amounts of neurotrophic factors during development. After peripheral nerve injury, the adult mammalian peripheral nervous system responds by making neurotrophic factors once again available, either by autocrine or paracrine sources. Three families of neurotrophic factors were compared, the neurotrophins, the GDNF family of neurotrophic factors, and the neuropoetic cytokines. Following a general overview of the mechanisms by which these neurotrophic factors mediate their effects, we reviewed the temporal pattern of expression of the neurotrophic factors and their receptors by axotomized motoneurons as well as in the distal nerve stump after peripheral nerve injury. We discussed recent experiments from our lab and others which have examined the role of neurotrophic factors in peripheral nerve injury. Although our understanding of the mechanisms by which neurotrophic factors mediate their effects in vivo are poorly understood, evidence is beginning to emerge that similar phenomena observed in vitro also apply to nerve regeneration in vivo.

Brauer, A. U., N. E. Savaskan, et al. (2003). "A new phospholipid phosphatase, PRG-1, is involved in axon growth and regenerative sprouting." Nat Neurosci 6(6): 572-8.
 Outgrowth of axons in the central nervous system is governed by specific molecular cues. Molecules detected so far act as ligands that bind to specific receptors. Here, we report a new membrane-associated lipid phosphate phosphatase that we have named plasticity-related gene 1 (PRG-1), which facilitates axonal outgrowth during development and regenerative sprouting. PRG-1 is specifically expressed in neurons and is located in the membranes of outgrowing axons. There, it acts as an ecto-enzyme and attenuates phospholipid-induced axon collapse in neurons and facilitates outgrowth in the hippocampus. Thus, we propose a novel mechanism by which axons are able to control phospholipid-mediated signaling and overcome the growth-inhibiting, phospholipid-rich environment of the extracellular space.

Brisby, H. (2003). "Nerve root injuries in patients with chronic low back pain." Orthop Clin North Am 34(2): 221-30.
 In conclusion, the nerve roots and the DRG play an important role in the pain mechanisms of patients suffering from chronic low back pain. Signs of demyelination and increased sensitization for stimuli occurs after a direct nerve root trauma, and the plasticity for the DRG also may change the response to a given peripheral stimuli when repeated frequently over a long period of time. The regeneration mechanisms of spinal nerve roots and DRG regarding function are slow, and the final grade of recurrence depends on the degree of injury. The limited regeneration mechanisms for nerve injury and the fact that "established chronic pain centers" are hard to influence after a long pain history favor an aggressive strategy for pain management. Today, a number of treatment strategies exist for chronic low back pain patients (with or without a diagnosed nerve root injury). These strategies include physiotherapy, nonsteroid anti-inflammatory drugs (NSAIDs), steroids, analgesics of different types and administration routes, surgery, and other sorts of invasive treatments. Further knowledge about the nerve root, DRG, and the rest of the nervous system in these patients is necessary; for understanding how and when to treat patients with chronic low back pain, we need to understand more about what we are trying to treat.

Bryant, S. M., K. L. Cumpston, et al. (2003). "Facial nerve neuritis secondary to ultraviolet radiation." Vet Hum Toxicol 45(4): 217-8.
 We describe a patient who developed facial nerve injury following significant exposure to UV radiation. A 49-y-old construction worker developed erythema and edema on the left side of his face (exposed side) 12 h after working within 18 in of a compromised metal halide incandescent light bulb for a total of 2 h. One month later, the patient noted a painful burning sensation over the left side of his face associated with marked left facial weakness and inability to close his eye (peripheral VIIth nerve palsy). Two months later, synkinetic left facial movements were consistent with aberrant regeneration. Over the next several months, forceful episodic spasmodic activity developed in the muscles of facial expression on the left, identical to that seen in hemifacial spasm. Rarely has UV radiation been implicated in damage to subcutaneous nerves. This case demonstrates that significant neurologic morbidity may follow high exposure to UV radiation.

Bryan, D. J., J. B. Tang, et al. (2003). "Enhanced peripheral nerve regeneration elicited by cell-mediated events delivered via a bioresorbable PLGA guide." J Reconstr Microsurg 19(2): 125-34.
 Using an established rat peripheral-nerve regeneration model, the authors have demonstrated enhancement of regeneration following subcutaneous priming of bioresorbable poly(lactic-co-glycolic)acid (PLGA) guides in vivo. Four weeks after nerve reconstruction, regeneration of the peripheral nerve through the cell-infiltrated guides displayed a significant increase in the total axon number and myelination status recorded in primed over unprimed guides, demonstrating the importance of cell-mediated events in the regeneration process. To define the different components enhancing nerve regeneration in this model, they have focused on identifying factors capable of eliciting Schwann-cell migration, since this has been identified as an early and necessary event in nerve regeneration. Using an in vitro migration assay, screening of a limited number of cellular and extracellular factors has demonstrated differential promotion of Schwann-cell migration. Of interest, combining fibronectin and bFGF resulted in a two-fold enhancement in Schwann-cell migration over that recorded with either alone. These results describe a rapid screening process for identifying various molecules and combinations thereof, with potential involvement in Schwann-cell migration. Coupling these findings to the use of the PLGA guide as an in vivo delivery system provides a rationale for the selection of exogenous factors to test for the enhancement of peripheral-nerve regeneration.

Buhusi, M., B. R. Midkiff, et al. (2003). "Close homolog of L1 is an enhancer of integrin-mediated cell migration." J Biol Chem 278(27): 25024-31.
 Close homolog of L1 (CHL1) is a member of the L1 family of cell adhesion molecules expressed by subpopulations of neurons and glia in the central and peripheral nervous system. It promotes neurite outgrowth and neuronal survival in vitro. This study describes a novel function for CHL1 in potentiating integrin-dependent cell migration toward extracellular matrix proteins. Expression of CHL1 in HEK293 cells stimulated their haptotactic migration toward collagen I, fibronectin, laminin, and vitronectin substrates in Transwell assays. CHL1-potentiated cell migration to collagen I was dependent on alpha1beta1 and alpha2beta1 integrins, as shown with function blocking antibodies. Potentiated migration relied on the early integrin signaling intermediates c-Src, phosphatidylinositol 3-kinase, and mitogen-activated protein kinase. Enhancement of migration was disrupted by mutation of a potential integrin interaction motif Asp-Gly-Glu-Ala (DGEA) in the sixth immunoglobulin domain of CHL1, suggesting that CHL1 functionally interacts with beta1 integrins through this domain. CHL1 was shown to associate with beta1 integrins on the cell surface by antibody-induced co-capping. Through a cytoplasmic domain sequence containing a conserved tyrosine residue (Phe-Ile-Gly-Ala-Tyr), CHL1 recruited the actin cytoskeletal adapter protein ankyrin to the plasma membrane, and this sequence was necessary for promoting integrin-dependent migration to extracellular matrix proteins. These results support a role for CHL1 in integrin-dependent cell migration that may be physiologically important in regulating cell migration in nerve regeneration and cortical development.

Burnett, A. L. (2003). "Neuroprotection and nerve grafts in the treatment of neurogenic erectile dysfunction." J Urol 170(2 Pt 2): S31-4; discussion S34.
 PURPOSE: The rationale for protecting the nerve supply of the penis derives mainly from the fact that neurological injury or disease states involving this organ commonly result in erectile dysfunction. Novel directions in the management of neurogenic erectile dysfunction that pertain specifically to sustaining penile neuronal function are described. MATERIALS AND METHODS: The review constitutes a summary of neuroprotective strategies for penile erection that are under investigation at the basic science level or have been brought to clinical practice. The basic exercise consisted primarily of a literature search using the National Library of Medicine PubMed Services, with references made to such keywords as nerve grafts, nerve growth factors, neuroprotection and nerve regeneration. RESULTS: Primary advances in this field have centered on repairing structural defects and restoring the functional integrity of the cavernous nerves of the penis. In the former autologous nerve conduits, such as sural nerve grafts, have been explored and used prominently in the context of radical prostatectomy. In the latter diverse neurotrophic treatments have been investigated, with progress mostly limited to animal models of cavernous nerve injury. Basic concepts and ongoing developments in the neurobiology of axonal regeneration were identified as being applicable to this area of neurourology. CONCLUSIONS: Because neurogenic origins represent a leading categorical cause of erectile dysfunction, the importance of developing and applying treatment approaches to alleviate neuropathic effects on the erectile tissue of the penis is certain. Medical and surgical innovations for preserving and reconstituting the functional nerve supply of the penis offer great promise in the management of erectile dysfunction.

Burnett, A. L. (2003). "Strategies to promote recovery of cavernous nerve function after radical prostatectomy." World J Urol 20(6): 337-42.
 While the application of penile autonomic nerve-sparing techniques during radical prostatectomy for clinically localized prostate cancer has improved erection recovery rates after surgery, many men still experience delayed or incomplete recovery of erectile function. In recognition of neuropathy as a likely basis for erectile dysfunction after radical prostatectomy, investigators have begun exploring new strategies to promote the functional recovery of nerves responsible for penile erection in the course of this management. Primary efforts continue for preserving the integrity of the penile nerves, while the next frontier in clinical management has encompassed strategies directed toward maximally restoring their function. Such strategies include cavernous nerve interposition grafting and neurotrophic treatments that meet nerve reconstructive and nerve regenerative objectives, respectively. Early successes with both innovations preclinically and clinically suggest their feasibility and potential roles to reduce the incidence of erectile dysfunction after radical prostatectomy. The purpose of this report is to review strategies under development to promote post-prostatectomy erectile function, particularly with respect to preserving penile innervation involved in this function.

Burnett, A. L. (2003). "Rationale for cavernous nerve restorative therapy to preserve erectile function after radical prostatectomy." Urology 61(3): 491-7.
 
Burrell, B. D., C. L. Sahley, et al. (2003). "Progressive recovery of learning during regeneration of a single synapse in the medicinal leech." J Comp Neurol 457(1): 67-74.
 The leech escape reflex-shortening of the body-can change with nonassociative conditioning, including sensitization, habituation, and dishabituation. Capacity for sensitization, which is an enhancement of the reflex, is lost when a single S-interneuron is ablated, but the reflex response itself remains. In the present experiments, the S-interneuron's axon in the living leech was filled with 6-carboxyfluorescein (6-CF) dye and cut with an argon laser microbeam (lambda = 488 nm). In contrast to sham-operated animals, axotomized preparations did not sensitize, reflecting the key role of the S-cell. By 2 weeks or more, S-cell axons had regenerated and reestablished synapses at their usual locations with neighboring S-cells. By 4 weeks, this restored the ability to sensitize to a level indistinguishable from that of controls, but an intermediate state of recovery was seen from 2-3 weeks after injury-a period not previously examined. The small capacity for sensitization among newly regenerated preparations was significantly lower than in sham controls but appeared higher than in animals whose cut S-cell axon had not regenerated its synapse. The results confirm the crucial role of the S-cell in sensitization. Moreover, full sensitization does not occur immediately upon synapse regeneration.

Buss, A., G. A. Brook, et al. (2003). "Gradual loss of myelin and formation of an astrocytic scar during Wallerian degeneration in the human spinal cord." Brain.
 Axons undergo Wallerian degeneration distal to a point of injury. Experimental investigations have documented many of the cellular and molecular events that underlie this behaviour. Since relatively little is known about such events in human CNS pathologies and current experimental intervention strategies indicate the possibility of significant axon regeneration along the original degenerated fibre tract, we performed an immunohistochemical investigation of the dynamics of Wallerian degeneration in post mortem spinal cords of patients who died 2 days to 30 years after either cerebral infarction or traumatic spinal cord injury. Neurofilament (NF) staining demonstrated a spatio-temporal pattern of axonal loss within degenerating descending nerve fibre tracts that could be detected close to the lesion as early as 12 days after injury and progressed to an almost complete loss of NF immunoreactivity at survival times of 1 year and longer. Immunohistochemistry for glial fibrillary acidic protein revealed a late astrocytic reaction starting at 4 months after injury in the degenerating tracts, leading to the long-term deposition of a dense astrocytic scar. These events were accompanied by the gradual reduction of myelin basic protein in affected nerve fibre tracts, leading to almost complete loss by 3 years after injury. Since the extracellular matrix molecule chondroitin sulphate proteoglycan (CSPG) is known to be strongly inhibitory for axonal regeneration and to be a major component of gliotic scar tissues, we investigated the possible deposition of CSPG within the degenerating nerve fibre tracts. Apart from a local up-regulation close to the lesion site, our results show no enhanced CSPG expression within degenerated tracts at any survival time. This suggests that despite the apparent lack of CSPG in Wallerian degeneration, the slow reduction of CNS myelin and the long-term deposition of a dense astrocytic scar may present an environment that is non-supportive for axon regrowth.

Buss, A. and M. E. Schwab (2003). "Sequential loss of myelin proteins during Wallerian degeneration in the rat spinal cord." Glia 42(4): 424-32.
 Axotomy of nerve fibers leads to the subsequent degeneration of their distal part, a process termed Wallerian degeneration (WD). While WD in the peripheral nervous system is usually followed by regeneration of the lesioned axons, central nervous system (CNS) neurons are generally unable to regrow. In this study, we investigated the process of WD in the dorsal columns of the rat spinal cord rostral to a mid-thoracic lesion. We confirm earlier studies describing a very delayed microglial and an early and sustained astroglial reaction finally leading to scar formation. Interestingly, we found a differential time course in the loss of myelin proteins depending on their location. Proteins situated on the periaxonal myelin membrane such as myelin associated glycoprotein disappeared early, within a few days after lesion, concomitantly with cytoskeletal axonal proteins, whereas compact myelin and outer myelin membrane proteins such as MBP and Nogo-A remained for long intervals in the degenerating tracts. Two distinct mechanisms are probably responsible for this difference: processes of protein destruction emanating from and initially probably located in the axon act on a time scale of 1-3 days. In contrast, the bulk of myelin destruction is due to phagocytosis known to be slow, prolonged, and inefficient in the CNS. These results may also have implications for future intervention strategies aiming at enhancing CNS regeneration.

Byers, M. R., H. Suzuki, et al. (2003). "Dental neuroplasticity, neuro-pulpal interactions, and nerve regeneration." Microsc Res Tech 60(5): 503-15.
 This review covers current information about the ability of dental nerves to regenerate and the role of tooth pulp in recruitment of regenerating nerve fibers. In addition, the participation of dental nerves in pulpal injury responses and healing is discussed, especially concerning pulp regeneration and reinnervation after tooth replantation. The complex innervation of teeth is highly asymmetric and guided towards specific microenvironments along blood vessels or in the crown pulp and dentin. Pulpal products such as nerve growth factor are distributed in the same asymmetric gradients as the dentinal sensory innervation, suggesting regulation and recruitment of those nerve fibers by those specific factors. The nerve fibers have important effects on pulpal blood flow and inflammation, while their sprouting and cytochemical changes after tooth injury are in response to altered pulpal cytochemistry. Thus, their pattern and neuropeptide intensity are indicators of pulp status, while their local actions continually affect that status. When denervated teeth are injured, either by pulp exposure on the occlusal surface or by replantation, they have more pulpal necrosis than occurs for innervated teeth. However, small pulp exposures on the side of denervated crowns or larger lesions in germ-free animals can heal well, showing the value of postoperative protection from occlusal trauma or from infection. Current ideas about dental neuroplasticity, neuro-pulpal interactions, and nerve regeneration are related to the overall topics of tooth biomimetics and pulp/dentin regeneration.

Cai, F. and C. J. Helke (2003). "Abnormal PI3 kinase/Akt signal pathway in vagal afferent neurons and vagus nerve of streptozotocin-diabetic rats." Brain Res Mol Brain Res 110(2): 234-44.
 The PI3 (phosphatidylinositol-3) kinase/Akt (protein kinase B) signal pathway is involved in the molecular signaling that regulates retrograde axonal transport of neurotrophins in the nervous system. Previous work showed that a reduced retrograde axonal transport of endogenous nerve growth factor (NGF) and neurotrophin-3 (NT-3) in the vagus nerve of diabetic rats occurred in the presence of normal production of neurotrophins and neurotrophin receptors. To assess the potential involvement of an impaired PI3 kinase/Akt signal pathway in the diabetes-induced reduction in retrograde axonal transport of neurotrophins in the vagus nerve, we characterized diabetes-induced changes in the PI3 kinase/Akt signal pathway in the vagus nerve and vagal afferent neurons. Control and streptozotocin (STZ)-induced diabetic rats with a duration of 16 weeks, kinase assays, Western blotting, and immunocytochemistry were used to show that diabetes resulted in alterations in activity and protein expression of the PI3 kinase/Akt signal pathway in the vagus nerve and vagal afferent neurons. Diabetes caused a significant decrease in enzymatic activity of PI3 kinase and Akt (52 and 36% of control, respectively) in the vagus nerve. The reduced enzymatic activity was not associated with decreased protein expression of the p85 subunit of PI3 kinase, Akt and phosphorylation of Akt (ser473). In contrast, there was a significant increase in the phosphorylation of p70s6 kinase (thr421/ser424) along with a normal protein expression of p70s6 kinase in the vagus nerve of diabetic rats. However, diabetes induced an overall decrease in immunoreactivity of the p85 subunit of PI3 kinase, phospho-Akt (ser473) and phospho-p70s6/p85s6 kinase (thr421/ser424) in vagal afferent neurons. Thus, impaired PI3 kinase/Akt signal pathway may partly account for the reduced retrograde axonal transport of neurotrophins in the vagus nerve of STZ-induced diabetic rats.

Calguner, E., R. Gozil, et al. (2003). "Atrophic and regenerative changes in rabbit mimic muscles after lidocaine and bupivacaine application." Anat Histol Embryol 32(1): 54-9.
 Destruction and denervation atrophy in skeletal muscles caused by the injection of local anaesthetics was investigated by injecting lidocaine or bupivacaine around the rabbit facial nerve to produce facial paralysis. Animals were then sacrificed at 2, 4, 6, and 8 weeks post-injection, and changes in mimic muscle tissue were assessed at each stage by light microscopy and electron microscopy. Atrophic changes were observed at 2-6 weeks after injection, and regeneration started at 6-8 weeks. Compared to bupivacaine, lidocaine caused more dramatic atrophic changes and was associated with slower muscle regeneration.

Calza, L., M. Fernandez, et al. (2003). "Nerve growth factor in the central nervous system: more than neuron survival." Arch Ital Biol 141(2-3): 93-102.
 
Cameron, A. A., G. Vansant, et al. (2003). "Identification of reciprocally regulated gene modules in regenerating dorsal root ganglion neurons and activated peripheral or central nervous system glia." J Cell Biochem 88(5): 970-85.
 Differential gene expression in the rat after injury of dorsal root ganglion neurons in vivo, and simulation injury of Schwann cells and oligodendrocytes in vitro was analyzed using high-density cDNA microarrays. The analyses were carried out to study the genetic basis of peripheral nerve regeneration, and to compare gene regulation in glia of the central (oligodendrocyte) and peripheral (Schwann cell) nervous systems. The genes showing significant differential regulation in the three study groups represented all aspects of cellular metabolism. However, two unexpected observations were made. Firstly, a number of identical genes were differentially regulated in activated Schwann cells, activated oligodendrocytes and regenerating DRG neurons. Specifically, a group of 113 out of 210 genes that were down-regulated in Schwann cells upon lipopolysaccharide (LPS) treatment, were identical to genes up-regulated in the injured, regenerating DRG. Furthermore, a group of 53 out of 71 genes that were down-regulated in interferon gamma (IFN-gamma)/LPS-activated oligodendrocytes, were identical to genes up-regulated in the DRG neurons. Finally, 22 genes were common to these three groups, i.e., down-regulated in activated oligodendrocytes, down-regulated in activated Schwann cells, and up-regulated in regenerating DRG neurons. Secondly, a group of 16 cell-cycle and proliferation-related genes were up-regulated in the DRG following sciatic nerve crush, despite the absence of cells undergoing mitosis in the DRG, or any significant presence of apoptosis-related gene expression. Therefore, it appears that in these three cell types, large sets of genes are reciprocally regulated upon injury and/or activation. This suggests that the activation of the injury-related gene expression program in cell derivatives of the neuroectoderm involves, in part, highly conserved genetic elements.

Campana, W. M., R. R. Myers, et al. (2003). "Identification of PINCH in Schwann cells and DRG neurons: shuttling and signaling after nerve injury." Glia 41(3): 213-23.
 Particularly interesting new cysteine-histidine rich protein (PINCH) is a double zinc finger domain (LIM)-only adapter protein that functions to recruit the integrin-linked kinase (ILK) to sites of integrin activation. Genetic studies have shown that PINCH and ILK are required for integrin signaling. Since integrin activation is associated with Schwann cell migration, neurite outgrowth and regeneration, this study examined PINCH in the normal peripheral nervous system and after chronic constriction injury (CCI) in adult Sprague-Dawley rats. Immunohistochemistry identified PINCH immunoreactivity in cell bodies of dorsal root ganglia (DRG) neurons, axons, satellite cells, and Schwann cells. PINCH immunostaining was localized to the membrane of uninjured DRG cell bodies consistent with its localization at a site of integrin activation. In contrast, 5 days following CCI, PINCH immunostaining was diffuse throughout the DRG cell cytoplasm. Confocal microscopy of primary and transformed Schwann cells localized PINCH in cytoplasmic, perinuclear and nuclear areas. Examination of the PINCH sequence revealed a putative leucine-rich nuclear export signal (NES) and an overlapping basic nuclear localization signal (NLS). To demonstrate nuclear export of PINCH, rabbit anti-PINCH IgG was microinjected into Schwann cell nuclei and allowed to combine with PINCH contained within the nucleus. Immunofluorescence showed that the PINCH and anti-PINCH IgG complex rapidly translocated to the cytoplasm. Treatment with leptomycin B caused nuclear accumulation of PINCH, indicating that the CRM1 pathway mediates nuclear export of PINCH. ILK activity in Schwann cells was enhanced by platelet-derived growth factor (PDGF) and tumor necrosis factor alpha. PINCH immunoprecipitates from PDGF- and TNFalpha-stimulated Schwann cells contained several high-molecular-weight threonine-phosphorylated proteins. Taken together, these results indicate that PINCH is an abundant shuttling/signaling protein in Schwann cells and DRG neurons.

Cao, X., J. Li, et al. (2003). "C3,4 transfer for neurotization of C5,6 nerve roots in brachial plexus injury in a rabbit model." J Reconstr Microsurg 19(4): 265-70.
 To evaluate the root neurotization properties of extraplexal donor nerves, an avulsion injury model of brachial plexus was created and repaired by C 3,4 nerve-root transfers in the rabbit. Eighteen rabbits were divided into three groups. In Group 1 (n = 6), the right C 5,6 nerve roots were avulsed and bridged by a nerve graft taken from the femoral nerve, with C 3,4 as C 3 to C 5 and C 4 to C 6. In Group 2 (n = 6), the right C 5,6 nerve roots were cut and directly sutured end-to-end. Group 3 (n = 6) was a negative group, in which C 5,6 nerve roots were avulsed without repair. All three groups were positively controlled by the contralateral side. Postoperative behavior observation and anatomic, electrophysiologic studies were conducted 4 months later for comparison among groups. Axon existence was observed by acetylcholinesterase staining. Results showed that active motion was not found in all three groups by the end of the study. Extraplexal nerve transfer indeed was able to re-neurotize the avulsed nerve roots down to their target organ, but C 3,4 nerve transfer was weaker than direct end-to-end suture, in terms of neurotization ability. The authors conclude that "root or trunk repair" for avulsion injury of the brachial plexus is possible, provided that the donor nerve has enough fibers and the nerve regeneration ability is increased by modern moleculobiologic techniques.

Carrasco, J., M. Penkowa, et al. (2003). "Role of metallothionein-III following central nervous system damage." Neurobiol Dis 13(1): 22-36.
 We evaluated the physiological relevance of metallothionein-III (MT-III) in the central nervous system following damage caused by a focal cryolesion onto the cortex by studying Mt3-null mice. In normal mice, dramatic astrogliosis and microgliosis and T-cell infiltration were observed in the area surrounding the lesioned tissue, along with signs of increased oxidative stress and apoptosis. There was also significant upregulation of cytokines/growth factors such as tumor necrosis factor-alpha, interleukin (IL)-1 alpha/beta, and IL-6 as measured by ribonuclease protection assay. Mt3-null mice did not differ from control mice in these responses, in sharp contrast to results obtained in Mt1- Mt2-null mice. In contrast, Mt3-null mice showed increased expression of several neurotrophins as well as of the neuronal sprouting factor GAP-43. Thus, unlike MT-I and MT-II, MT-III does not affect the inflammatory response elicited in the central nervous system by a cryoinjury, nor does it serve an important antioxidant role, but it may influence neuronal regeneration during the recovery process.

Carro, E., J. L. Trejo, et al. (2003). "Brain repair and neuroprotection by serum insulin-like growth factor I." Mol Neurobiol 27(2): 153-62.
 The existence of protective mechanisms in the adult brain is gradually being recognized as an important aspect of brain function. For many years, self-repair processes in the post-embryonic brain were considered of minor consequence or nonexistent. This notion dominated the study of neurotrophism. Thus, although the possibility that neurotrophic factors participate in brain function in adult life was prudently maintained, the majority of the studies on the role of trophic factors in the brain were focused on developmental aspects. With the recent recognition that the adult brain keeps a capacity for cell renewal, although limited, a new interest in the regenerative properties of brain tissue has emerged. New findings on the role of insulin-like growth factor I (IGF-I), a potent neurotrophic peptide present at high levels in serum, may illustrate this current trend. Circulating IGF-I is an important determinant of proper brain function in the adult. Its pleiotropic effects range from classical trophic actions on neurons such as housekeeping or anti-apoptotic/ pro-survival effects to modulation of brain-barrier permeability, neuronal excitability, or new neuron formation. More recent findings indicate that IGF-I participates in physiologically relevant neuroprotective mechanisms such as those triggered by physical exercise. The increasing number of neurotrophic features displayed by serum IGF-I reinforces the view of a physiological neuroprotective network formed by IGF-I, and possibly other still uncharacterized signals. Future studies with IGF-I, and hopefully other neurotrophic factors, will surely reveal and teach us how to potentiate the self-reparative properties of the adult brain.

Ceballos, D., N. Lago, et al. (2003). "Role of metallothioneins in peripheral nerve function and regeneration." Cell Mol Life Sci 60(6): 1209-16.
 The physiological role of the metallothionein (MT) family of proteins during peripheral nerve injury and regeneration was examined in Mt1+ 2 and Mt3 knockout (KO) mice. To this end, the right sciatic nerve was crushed, and the regeneration distance was evaluated by the pinch test 2-7 days postlesion (dpl) and electrophysiologically at 14 dpl. The quality of the regeneration was assessed by light microscopy and immunohistochemical methods. The results show that the regeneration distance was greater in the Mt3 KO than in the Mt1+ 2 KO mice, whereas control mice showed intermediate values. Moreover, the number of regenerating axons in the distal tibial nerve was significantly higher in Mt3KO mice than in the other two strains at 14 dpl. Immunoreactive profiles to protein gene product 9.5 were present in the epidermis and the sweat glands of the plantar skin of the hindpaw of the Mt3 KO group. The improved regeneration observed with the Mt3 KO mice was confirmed by compound nerve action potentials that were recorded from digital nerves at 14 dpl only in this group. We conclude that Mt3 normally inhibits peripheral nerve regeneration.

Chafik, D., D. Bear, et al. (2003). "Optimization of Schwann cell adhesion in response to shear stress in an in vitro model for peripheral nerve tissue engineering." Tissue Eng 9(2): 233-41.
 The design of nerve guidance channels (NGCs) is evolving to produce a favorable environment for neural regeneration. We created an in vitro model to evaluate the interactions between three centrally important components of this altered host environment: (1). Schwann cells, (2). substrate, and (3). sustained mechanical stimulus in the form of shear stress with laminar fluid flow. Preconfluent Schwann cells were plated on slides coated either with laminin, poly-D-lysine, type IV collagen, or fibronectin. These slides were placed into custom-designed, parallel-plate, flow chambers and were administered laminar fluid flow at a rate of 15 mL/min for 2 h. Schwann cell adhesion assays demonstrated that laminin (mean, 86.1%; SEM, 4.47%) and fibronectin (mean, 81.7%; SEM, 3.24%) were statistically superior to collagen type IV (mean, 57.7%; SEM, 3.96%) and poly-D-lysine (mean, 58.0%; SEM, 4.97%) (p < 0.001). Fibronectin (mean, 12.20%; SEM, 0.374%) induced statistically greater Schwann cell proliferation than did laminin (mean, 8.14%; SEM, 0.682%) (p < 0.001). Therefore, we recommend that fibronectin should be used as an important component of NGCs with further in vivo studies. As mechanical stress is an integral part of the host environment, our study is the first to incorporate this factor into an in vitro model for peripheral nerve tissue engineering.

Chalfoun, C., T. Scholz, et al. (2003). "Primary nerve grafting: A study of revascularization." Microsurgery 23(1): 60-5.
 It was the purpose of this study to evaluate the revascularization of primary nerve repair and grafts using orthogonal polarization spectral (OPS) (Cytometrix, Inc.) imaging, a novel method for real-time evaluation of microcirculatory blood flow. Twenty male Sprague Dawley rats (250 g) were anesthetized with vaporized halothane and surgically prepared for common peroneal nerve resection. Group I animals (n = 10) underwent primary neurorraphy following transection, utilizing a microsurgical technique with 10-0 nylon suture. Group II (n = 10) animals had a 7-mm segment of nerve excised, reversed, and subsequently replaced as a nerve graft under similar techniques. All animals were evaluated using the OPS imaging system on three portions (proximal, transection site/graft, and distal) of the nerve following repair or grafting. Reevaluation of 5 animals randomly selected from each group using the OPS imaging system was again performed on days 14 and 28 following microsurgical repair/grafting. Values were determined by percent change in vascularity of the common peroneal nerve at 0 hr following surgery. Real-time evaluation of blood flow was utilized as an additional objective criterion. Percent vascularity in group I and II animals increased from baseline in all segments at day 14. By day 28, vascularity in nerves of group I rats decreased in all segments to values below baseline, with the exception of the transection site, which remained at a higher value than obtained directly after surgical repair. In group II animals, vascularity remained above baseline in all segments except the distal segment, which returned to vascularity levels similar to those at 0 hr. Further, occlusion of the vessels demonstrated in the graft and distal segments following initial transection appeared to be corrected. This study suggests that revascularization may occur via bidirectional inosculation with favored proximal vascular growth advancement. The use of real-time imaging offers a unique evaluation of tissues through emerging technologies.

Chan, Y. M., L. W. Yick, et al. (2003). "Inhibition of caspases promotes long-term survival and reinnervation by axotomized spinal motoneurons of denervated muscle in newborn rats." Exp Neurol 181(2): 190-203.
 We examined whether (1) a pan-caspase inhibitor, Boc-D-FMK, exerts long-term neuroprotective effects on spinal motoneurons (MNs) after root avulsion in neonatal rats and (2) whether the rescued spinal MNs regenerate their axons into a peripheral nerve (PN) graft and reinnervate a previously denervated target muscle. Eight weeks after root avulsion, 67% of spinal MNs remained in the Boc-D-FMK-treated group, whereas all MNs died in the sham control group. By 12 weeks postinjury, however, all Boc-D-FMK treated MNs died. In the regeneration experiment, a PN graft was implanted at different times after injury. The animals were allowed to survive for 4 weeks following the operation. Without caspase inhibition, MNs did not regenerate at any time point. In animals treated with Ac-DEVD-CHO, a caspase-3-specific inhibitor, and Boc-D-FMK, 44 and 62% of MNs, respectively, were found to regenerate their axons into a PN graft implanted immediately after root avulsion. When the PN graft was implanted 2 weeks after injury, however, MNs failed to regenerate following Ac-DEVD-CHO treatment, whereas 53% of MNs regenerated their axons into the graft after treatment with Boc-D-FMK. No regeneration was observed when a PN graft was implanted later than 2 weeks after injury. In the reinnervation study, injured MNs and the target biceps muscle were reconnected by a PN bridge implanted 2 weeks after root avulsion with administration of Boc-D-FMK. Eight weeks following the operation, 39% of MNs reinnervated the biceps muscle. Morphologically normal synapses and motor endplates were reformed in the muscle fibers. Collectively, these data provide evidence that injured neonatal motoneurons can survive and reinnervate peripheral muscle targets following inhibition of caspases.

Chao, M. V. (2003). "Neurotrophins and their receptors: a convergence point for many signalling pathways." Nat Rev Neurosci 4(4): 299-309.
 
Chavez-Delgado, M. E., J. Mora-Galindo, et al. (2003). "Facial nerve regeneration through progesterone-loaded chitosan prosthesis. A preliminary report." J Biomed Mater Res 67B(2): 702-11.
 Biodegradable nerve guides have represented new treatment alternatives for nerve repairing. They are gradually biodegradable, exert biological effects directly to the injured nerve, and act as drug- or cell-delivery devices. Furthermore, progesterone (PROG) has been demonstrated to promote injured peripheral nerve regeneration. In this study, it was hypothesized that PROG delivered from chitosan prostheses provides better facial nerve regenerative response than chitosan prostheses with no PROG. As there are no reports on the use of the former as nerve-guide material in the regeneration of injured nerves, this is the main objective of the present work. Chitosan prostheses containing PROG were used to bridge 10-mm gaps in rabbit facial nerves. The regenerated nerves were evaluated 45 days after implantation in animals with the use of light microscopy and morphometric analysis. Gas chromatography was used in order to quantify PROG content in prosthesis prior to and after implantation in subcutaneous tissue at different periods of up to 60 days. In addition, the prosthesis walls were evaluated with histological techniques in order to assess their integrity and the surrounding tissue reaction. Chitosan prostheses allowed PROG release during the time needed for nerve regeneration. At 45 days myelinated nerve fibers were observed in both the proximal and distal stumps. This parameter and the N ratio were higher in the progesterone-treated group when compared to that of the vehicle control. Findings indicate that chitosan prostheses were useful in nerve regeneration, acting as a long-lasting PROG delivery device a faster nerve regeneration.

Chen, Z. L. and S. Strickland (2003). "Laminin gamma1 is critical for Schwann cell differentiation, axon myelination, and regeneration in the peripheral nerve." J Cell Biol 163(4): 889-99.
 Laminins are heterotrimeric extracellular matrix proteins that regulate cell viability and function. Laminin-2, composed of alpha2, beta1, and gamma1 chains, is a major matrix component of the peripheral nervous system (PNS). To investigate the role of laminin in the PNS, we used the Cre-loxP system to disrupt the laminin gamma1 gene in Schwann cells. These mice have dramatically reduced expression of laminin gamma1 in Schwann cells, which results in a similar reduction in laminin alpha2 and beta1 chains. These mice exhibit motor defects which lead to hind leg paralysis and tremor. During development, Schwann cells that lack laminin gamma1 were present in peripheral nerves, and proliferated and underwent apoptosis similar to control mice. However, they were unable to differentiate and synthesize myelin proteins, and therefore unable to sort and myelinate axons. In mutant mice, after sciatic nerve crush, the axons showed impaired regeneration. These experiments demonstrate that laminin is an essential component for axon myelination and regeneration in the PNS.

Chen, J., Y. Tu, et al. (2003). "Heme oxygenase-1 and heme oxygenase-2 have distinct roles in the proliferation and survival of olfactory receptor neurons mediated by cGMP and bilirubin, respectively." J Neurochem 85(5): 1247-61.
 Heme oxygenase (HO) is implicated in protection against oxidative stress, proliferation and apoptosis in many cell types, including neurons. We utilized olfactory receptor neurons (ORNs) as a model to define the roles of HO-1 and HO-2 in neuronal development and survival, and to determine the mediators of these effects. The olfactory system is a useful model as ORNs display neurogenesis post-natally and do not contain nitric oxide synthase (NOS) activity, which could confound results. HO isoforms were expressed in ORNs during embryogenesis and post-natally. Mice null for either HO-1 or HO-2 displayed decreased proliferation of neuronal precursors. However, apoptosis was increased only in HO-2 null mice. Cyclic GMP immunostaining was reduced in ORNs in both genotypes, providing direct evidence that HO mediates cGMP production in vivo. Bilirubin immunostaining was reduced only in HO-2 null mice. These roles for HO-1 and HO-2 were confirmed using detergent ablation of the epithelium to observe increased neurogenesis of ORNs after target disruption in HO null mice. Primary cultures of ORNs revealed that proliferative and survival effects of HO were mediated through cGMP and bilirubin, respectively. These results support a role for HO, the CO-cGMP signaling system and bilirubin in neurodevelopment and in response to injury.

Chen, P., F. Zindy, et al. (2003). "Progressive hearing loss in mice lacking the cyclin-dependent kinase inhibitor Ink4d." Nat Cell Biol 5(5): 422-6.
 Maintenance of the post-mitotic state in the post-natal mammalian brain is an active process that requires the cyclin-dependent kinase inhibitors (CKIs) p19Ink4d (Ink4d) and p27Kip1 (Kip1). In animals with targeted deletions of both Ink4d and Kip1, terminally differentiated, post-mitotic neurons are observed to re-enter the cell cycle, divide and undergo apoptosis. However, when either Ink4d or Kip1 alone are deleted, the post-mitotic state is maintained, suggesting a redundant role for these genes in mature neurons. In the organ of Corti--the auditory sensory epithelium of mammals--sensory hair cells and supporting cells become post-mitotic during embryogenesis and remain quiescent for the life of the animal. When lost as a result of environmental insult or genetic abnormality, hair cells do not regenerate, and this loss is a common cause of deafness in humans. Here, we report that targeted deletion of Ink4d alone is sufficient to disrupt the maintenance of the post-mitotic state of sensory hair cells in post-natal mice. In Ink4d-/- animals, hair cells are observed to aberrantly re-enter the cell cycle and subsequently undergo apoptosis, resulting in progressive hearing loss. Our results identify a novel mechanism underlying a non-syndromic form of progressive hearing loss in mice.

Chen, Y. and R. A. Swanson (2003). "Astrocytes and brain injury." J Cereb Blood Flow Metab 23(2): 137-49.
 Astrocytes are the most numerous cell type in the central nervous system. They provide structural, trophic, and metabolic support to neurons and modulate synaptic activity. Accordingly, impairment in these astrocyte functions during brain ischemia and other insults can critically influence neuron survival. Astrocyte functions that are known to influence neuronal survival include glutamate uptake, glutamate release, free radical scavenging, water transport, and the production of cytokines and nitric oxide. Long-term recovery after brain injury, through neurite outgrowth, synaptic plasticity, or neuron regeneration, is influenced by astrocyte surface molecule expression and trophic factor release. In addition, the death or survival of astrocytes themselves may affect the ultimate clinical outcome and rehabilitation through effects on neurogenesis and synaptic reorganization.

Cheng, M., W. Cao, et al. (2003). "Studies on nerve cell affinity of biodegradable modified chitosan films." J Biomater Sci Polym Ed 14(10): 1155-67.
 Chitosan, a natural polysaccharide that has excellent biocompatibility and biodegradability, can be used as nerve conduit material. The purpose of this work was to study the ability of chitosan and some chitosan-derived materials to facilitate nerve cell attachment, differentiation and growth. The biomaterials studied were chitosan, poly-L-lysine-blended chitosan (CP), collagen-blended chitosan (CC) and albumin-blended chitosan (CA), with collagen control material. Culture of PC12 cells and fetal mouse cerebral cortex (FMCC) cells on these biomaterials was used to evaluate their nerve cell affinity. The composite materials, including CP, CC and CA, had significantly improved nerve cell affinity compared to chitosan, as established by increasing attachment, differentiation and growth of PC12 cells. FMCC cells could also grow better on composite materials than on chitosan. CP exhibited the best nerve cell affinity among these three types of composite material. CP is an even better material in promoting neurite outgrowth than collagen, a substrate that is widely used in tissue engineering, suggesting that CP is a promising candidate material for nerve regeneration.

Cheng, M., J. Deng, et al. (2003). "Study on physical properties and nerve cell affinity of composite films from chitosan and gelatin solutions." Biomaterials 24(17): 2871-80.
 A series of chitosan-gelatin composite films was prepared by varying the ratio of constituents. FT-IR and X-ray analysis showed good compatibility between these two biopolymers. Differential scanning calorimetry (DSC) analysis indicated that the water take-up of chitosan film increased when blended with gelatin. Composite film exhibited a lower Young's modulus and a higher percentage of elongation-at-break compared with chitosan film, especially in wet state. All composite films were hydrophilic materials with water contact angles ranging from 55 degrees to 65 degrees. The results obtained from ELISA indicated the adsorption amount of fibronectin on composite films was much higher than on chitosan film. PC12 cells culture was used to evaluate the nerve cell affinity of materials. The cells cultured on the composite film with 60wt% gelatin differentiated more rapidly and extended longer neurites than on chitosan film. The results suggest that the soft and elastic complex of chitosan and gelatin, which has better nerve cell affinity compared to chitosan, is a promising candidate biomaterial for nerve regeneration.

Choi, D. and G. Raisman (2003). "Immune rejection of a facial nerve xenograft does not prevent regeneration and the return of function: an experimental study." Neuroscience 121(2): 501-7.
 Nerve grafts may be used to repair damaged peripheral nerves and also to facilitate spinal cord regeneration after experimental trauma. Little is known, however, about the possible use of xenografts and the role of immune rejection in the outcome of repair. In rats, excision of a short (7-8 mm) segment of facial nerve at its exit point from the skull base results in a permanent deficit in eye closure in the blink reflex. This deficit can be repaired by transplantation of a segment of either syngeneic rat facial nerve or xenogeneic Balb-C mouse sciatic nerve either with or without cyclosporine immunosuppression. With longer (15-20 mm) transplants, however, restoration of eye closure becomes dependent on cyclosporine administration. Thus, in a situation where nerve repair does not occur without a graft, a host immune attack has an attritional effect which is not sufficient to prevent repair over short distances, but becomes obvious when the regenerating fibres have to cross longer segments of transplanted tissue.

Christianson, J. A., J. T. Riekhof, et al. (2003). "Restorative effects of neurotrophin treatment on diabetes-induced cutaneous axon loss in mice." Exp Neurol 179(2): 188-99.
 Chronic hyperglycemia in diabetes causes a variety of somatosensory deficits, including reduced cutaneous innervation of distal extremities. Deficient neurotrophin support has been proposed to contribute to the development of diabetic neuropathy. Here, studies were carried out in streptozotocin (STZ)-treated mice to determine whether (1) cutaneous innervation deficits develop in response to hyperglycemia, (2) neurotrophin production is altered in the skin, and (3) neurotrophin treatment improves cutaneous innervation deficits. Cutaneous innervation was quantified in the hindlimb skin using antibodies that label nerve growth factor- (NGF) responsive (CGRP), glial cell line-derived neurotrophic factor (GDNF)/neurturin (NTN) -responsive (P2X(3)), or all cutaneous axons (PGP 9.5). Diabetic mice displayed severely reduced cutaneous innervation for all three antibodies in both flank and footpad skin regions, similar to reports of cutaneous innervation loss in human diabetic patients. Qualitative assessment of mRNAs for NGF, GDNF, and NTN demonstrated that these mRNAs were expressed in hindlimb flank and footpad skin from diabetic mice. Next, diabetic mice were then treated intrathecally for 2 weeks with NGF, GDNF, or NTN. NGF treatment failed to improve cutaneous innervation, but stimulated axon branching. In comparison, GDNF and NTN treatment increased cutaneous innervation and axon branching. Our results reveal that similar to human diabetic patients, STZ-induced diabetes significantly reduces hindlimb cutaneous innervation in mice. Importantly, intrathecal treatment using GDNF or NTN strongly stimulated axon growth and branching, suggesting that administration of these trophic factors can improve cutaneous innervation deficits caused by diabetes.

Colman, D., C. Lubetzki, et al. (2003). "Multiple paths towards repair in multiple sclerosis." Trends Neurosci 26(2): 59-61.
 Multiple sclerosis (MS) is an inflammatory demyelinating disease of the CNS, affecting approximately 1/1000 individuals in the Western world. Available treatments limit CNS inflammation and strategies to repair damage in the CNS offer the potential of recovery of both tissue and function. With further fundamental knowledge developing, this area is ripe for 'translation' to clinical application.

Constantinidis, J., A. Akbarian, et al. (2003). "Effects of immediate and delayed facial-facial nerve suture on rat facial muscle." Acta Otolaryngol 123(8): 998-1003.
 OBJECTIVE: To investigate regeneration of the mimic musculature after delayed facial nerve repair. MATERIALS AND METHOD: In 30 rats the facial nerve on the right side was resected and immediately repaired with an end-to-end anastomosis. The entire levator labii muscle was removed on the right side and histochemically and morphometrically analyzed at 7, 14, 21, 28, 90 and 180 days after immediate nerve repair. In a further 20 rats, reinnervation was performed after 4 weeks in a further operation and the muscle was assessed at 14, 28, 90 and 180 days. Seven muscles of four normal rats were used as controls. RESULTS: Three muscle fiber types could be identified by comparative histochemical analysis of two enzymes of succinate dehydrogenase (SDH) and m-ATPase: oxidative glycolytic (FOG) fibers, fast glycolytic (FG) fibers and intermediate muscle fibers that were designated SDH-INT. The number of fibers and the frequency distribution of single-fiber cross-sectional areas were determined for each type of fiber. No significant difference in the cross-sectional area of all fiber types was noted after 180 days when comparing immediate and delayed end-to-end sutures. In contrast, comparison of the fiber-type composition showed a significant increase in the number of FG fibers in the delayed suture, at the expense of FOG fibers. After immediate reinnervation the muscle approximately attained its natural fiber-type composition, whereas after delayed reinnervation the muscle showed an overall loss of oxidative capacity and differentiation towards phasic activity. CONCLUSION: Changes in fiber-type composition following delayed nerve suture are irreversible and have specific effects on the precise function of the muscle. Immediate nerve repair improves axonal reinnervation.

Corey, J. M. and E. L. Feldman (2003). "Substrate patterning: an emerging technology for the study of neuronal behavior." Exp Neurol 184 Suppl 1: S89-96.
 Extracellular matrix (ECM) proteins and cell-cell adhesion molecules (CAM) play important roles in neuronal development and differentiation. In the investigation of these roles, patterned substrates have proven to be a notably useful tool. Photolithographic and microprinting techniques can be used to make patterns of ECMs, CAMs, amino acids, and organofunctional groups for culturing neurons and other cell types. Experiments performed using these substrates have provided unique insights into the roles of cell-substratum adhesion, cell shape, and ECM composition on important cell functions, including survival, migration, neurite outgrowth, and development of polarity. Patterns may also be designed to localize cell bodies and confine their processes to predetermined areas of a substrate. Finally, the behavior of neurons on patterned substrates may prove helpful in the design of scaffoldings and nerve guides tailored for regeneration and repair of the nervous system.

Corti, S., F. Locatelli, et al. (2003). "Neuronal generation from somatic stem cells: current knowledge and perspectives on the treatment of acquired and degenerative central nervous system disorders." Curr Gene Ther 3(3): 247-72.
 Stem cell transplantation through cell replacement or as vector for gene delivery is a potential strategy for the treatment of neurodegenerative diseases. Several studies have reported the transdifferentiation of different somatic stem cells into neurons in vitro or after transplantation into animal models. This observation has pointed out the perspective of using an ethical and accessible cell source to "replace" damaged neurons or provide support to brain tissue. However, recent findings such as the cell fusion phenomenon have raised some doubts about the real existence of somatic stem cell plasticity. In this review, we will discuss current evidence and controversial issues about the neuroneogenesis from various sources of somatic cells focusing on the techniques of isolation, expansion in vitro as well as the inductive factors that lead to transdifferentiation in order to identify the factors peculiar to this process. The morphological, immunochemical, and physiological criteria to correctly judge whether the neuronal transdifferentation occurred are critically presented. We will also discuss the transplantation experiments that were done in view of a possible clinical therapeutic application. Animal models of stroke, spinal cord and brain trauma have improved with Mesenchymal Stem Cells or Bone Marrow transplantation. This improvement does not seem to depend on the replacement of the lost neurons but may be due to increased expression levels of neurotrophic factors, thus suggesting a beneficial effect of somatic cells regardless of transdifferentiation. Critical understanding of available data on the mechanisms governing the cell fate reprogramming is a necessary achievement toward an effective cell therapy.

Cuadros-Romero, M. (2003). "Nerve regeneration using polyglicolic acid as alternative to grafting." J Hand Surg [Am] 28 Suppl 1: 17.
 
Csoknya, M., J. Barna, et al. (2003). "Reorganization of monoaminergic systems in the earthworm, Eisenia fetida, following brain extirpation." J Exp Zoolog Part A Comp Exp Biol 296(1): 18-29.
 The present study describes the major aspects of how monoaminergic (serotonin, dopamine) systems change in the course of regeneration of the brain in the earthworm (Eisenia fetida), investigated by immunocytochemistry, HPLC assay, and ligand binding. Following brain extirpation, the total regeneration time is about 80 days at 10 degrees C. On the 3rd postoperative day serotonin, and on the 11th postoperative day tyrosine hydroxylase-immunoreactive neurons can be observed in the wound tissue. Thereafter the number of the immunoreactive cells increases gradually, and by the 76th-80th postoperative days all serotonin- and tyrosine hydroxylase-immunopositive neurons can be found in their final positions, similarly to those observed in the intact brain. Labeled neurons located in the dorsal part of the regenerated brain appear earlier than the cells in lateral and ventral positions. Both serotonin- and tyrosine hydroxylase-immunoreactive neurons of the newly formed brain seem to originate from undifferentiated neuroblasts situated within and around the ventral ganglia and the pleura. Dopaminergic (tyrosine hydroxylase-immunoreactive) elements may additionally derive from the proliferation of neurons localized in the subesophageal ganglion and the pharyngeal nerve plexus. Following brain extirpation, both serotonin and dopamine levels, assayed by HPLC, first increase in the subesophageal ganglion; by the 25th day of regeneration, the monoamine content decreases in it and increases in the brain. Hence it is suggested that monoamines are at least partly transported from this ganglion to the regenerating brain. At the same time, (3)H-LSD binding can be detected in the regenerating brain from the 3rd postoperative day, showing a continuous increase until the 80th postoperative day, suggesting a guiding role of postsynaptic elements in the monoaminergic reinnervation of the newly formed brain.

Cui, S. S., C. P. Yang, et al. (2003). "Valproic acid enhances axonal regeneration and recovery of motor function after sciatic nerve axotomy in adult rats." Brain Res 975(1-2): 229-36.
 It has recently been demonstrated that valproic acid (VPA) robustly promotes neurite outgrowth, activates the extracellular signal regulated kinase pathway, and increases growth cone-associated protein 43 and bcl-2 levels in cultured human neuroblastoma SH-SY5Y cells. We hypothesized that VPA could also enhance peripheral nerve regeneration in adult animals. To test this hypothesis, we examined the effects of VPA (300 mg/kg daily for 16 weeks) on sciatic axonal regeneration following single or conditional axotomies in rats. The results showed that in VPA-treated rats there was a significant increase in the total numbers of regenerated myelinated nerve fibers and reinnervated muscle fibers in comparison with those rats not treated with VPA. As measured by sciatic function index and toe spread index, the motor function of the reinnervated hind limbs of rats receiving single axotomy without VPA treatment significantly improved at week 8 and reached plateau levels at about week 11, whereas the motor function of the reinnervated hind limbs of rats receiving single axotomy plus VPA and rats receiving conditional axotomy with or without VPA treatment significantly improved at week 4 and reached plateau levels at about week 8; there was no significant difference of the motor function among the three later groups. The results demonstrated that VPA is able to enhance sciatic nerve regeneration and recovery of motor function in adult rats, suggesting the potential clinical application of VPA for the treatment of peripheral nerve injury in humans.

Cui, Q., M. A. Pollett, et al. (2003). "A new approach to CNS repair using chimeric peripheral nerve grafts." J Neurotrauma 20(1): 17-31.
 We have examined whether transplanted freeze-thawed peripheral nerve (PN) sheaths repopulated ex vivo with purified adult Schwann cells (SCs) support the regeneration of adult rat retinal ganglion cell (RGC) axons. Cultured adult SCs were derived from donor rats or from the host animals themselves. We also transplanted PN sheaths filled with neonatal SCs or donor adult olfactory ensheathing glia (OEG). 100,000 cells were injected into 1.5-cm lengths of freeze-thawed PN. After 2 days in culture, repopulated PN segments were grafted onto the transected optic nerve of adult Fischer rats. Three weeks later, 6% fluorogold (FG) was applied to distal PN. Retrogradely labeled RGCs were counted in retinal wholemounts and PN grafts were processed for immunohistochemistry. As expected, there was no RGC axon regeneration in cell-free grafts. Regrowth was also absent in neonatal SC- and adult OEG-filled grafts, which contained only small numbers of surviving donor cells. Many cells were, however, seen in adult SC repopulated PN grafts, intermingled with pan-neurofilament(+) and GAP-43(+) fibers. SCs were aligned along the grafts and were S-100(+), p75(+). Ultrastructurally, SCs were associated with myelinated and unmyelinated axons. Hundreds of FG-labeled RGCs were seen in retinas of rats with congeneic or allogeneic PN sheaths repopulated with either donor or autologous (host-derived) adult SCs. Intraocular CNTF injections significantly increased the number of regenerating RGCs in donor and autologous adult SC groups. The use of chimeric grafts to bridge CNS tissue defects could provide a clinical alternative to using multiple PN autografts, the harvesting of which would exacerbate peripheral dysfunction in already injured patients.

Cui, Q., H. K. Yip, et al. (2003). "Intraocular elevation of cyclic AMP potentiates ciliary neurotrophic factor-induced regeneration of adult rat retinal ganglion cell axons." Mol Cell Neurosci 22(1): 49-61.
 In vitro, cyclic AMP (cAMP) elevation alters neuronal responsiveness to diffusible growth factors and myelin-associated inhibitory molecules. Here we used an established in vivo model of adult central nervous system injury to investigate the effects of elevated cAMP on neuronal survival and axonal regeneration. We studied the effects of intraocular injections of neurotrophic factors and/or a cAMP analogue (CPT-cAMP) on the regeneration of axotomized rat retinal ganglion cell (RGC) axons into peripheral nerve autografts. Elevation of cAMP alone did not significantly increase RGC survival or the number of regenerating RGCs. Ciliary neurotrophic factor increased RGC viability and axonal regrowth, the latter effect substantially enhanced by coapplication with CPT-cAMP. Under these conditions over 60% of surviving RGCs regenerated their axons. Neurotrophin-4/5 injections also increased RGC viability, but there was reduced long-distance axonal regrowth into grafts, an effect partially ameliorated by cAMP elevation. Thus, cAMP can act cooperatively with appropriate neurotrophic factors to promote axonal regeneration in the injured adult mammalian central nervous system.

Curtis, M. A., E. B. Penney, et al. (2003). "Increased cell proliferation and neurogenesis in the adult human Huntington's disease brain." Proc Natl Acad Sci U S A 100(15): 9023-7.
 Neurogenesis has recently been observed in the adult human brain, suggesting the possibility of endogenous neural repair. However, the augmentation of neurogenesis in the adult human brain in response to neuronal cell loss has not been demonstrated. This study was undertaken to investigate whether neurogenesis occurs in the subependymal layer (SEL) adjacent to the caudate nucleus in the human brain in response to neurodegeneration of the caudate nucleus in Huntington's disease (HD). Postmortem control and HD human brain tissue were examined by using the cell cycle marker proliferating cell nuclear antigen (PCNA), the neuronal marker beta III-tubulin, and the glial cell marker glial fibrillary acidic protein (GFAP). We observed a significant increase in cell proliferation in the SEL in HD compared with control brains. Within the HD group, the degree of cell proliferation increased with pathological severity and increasing CAG repeats in the HD gene. Most importantly, PCNA+ cells were shown to coexpress beta III-tubulin or GFAP, demonstrating the generation of neurons and glial cells in the SEL of the diseased human brain. Our results provide evidence of increased progenitor cell proliferation and neurogenesis in the diseased adult human brain and further indicate the regenerative potential of the human brain.

Daniele, H. R. and L. Aguado (2003). "Early compensatory sensory re-education." J Reconstr Microsurg 19(2): 107-10; discussion 111-2.
 After a neurorrhaphy, there will be a distal disconnection between the cortex and skin receptors, along with interruption of sensibility information. This report demonstrates the efficacy of a new sensory re-education program for achieving optimal sensation in a relatively short time. Between 1999 and 2001, in the authors' Hand Rehabilitation Department, 11 patients with previous neurorrhaphy were subjected to a program of early "compensatory sensory re-education." Lesions were caused by clean cut. There were 13 primary digital nerve procedures, 12 at the distal palmar MP level, and one at the radial dorsal branch of the index (just after emerging from the common digital nerve). The technique of compensatory sensory re-education was based on a previous, but modified, sensory re-education method. In order to evaluate the results in the compensatory sensory re-education series described, additional tests for evaluation of achieved functional sensibility were used. The authors' best results were achieved in a maximum of 8 weeks (4-8 weeks), much less time than with the original method (1-2 years). Using the British classification, it was possible to compare the achieved levels of sensibility and the time required for optimal results. The different methods of sensibility re-education may be similar, but with the authors' compensatory sensory re-education method, substantial time is saved.

David, S. and S. Lacroix (2003). "Molecular approaches to spinal cord repair." Annu Rev Neurosci 26: 411-40.
 Axon growth inhibitors associated with myelin and the glial scar contribute to the failure of axon regeneration in the injured adult mammalian central nervous system (CNS). A number of these inhibitors, their receptors, and signaling pathways have been identified. These inhibitors can now be neutralized by a variety of approaches that point to the possibility of developing new therapeutic strategies to stimulate regeneration after spinal cord injury.

de Buys Roessingh, A. S. and O. Reinberg (2003). "Open or closed pinning for distal humerus fractures in children?" Swiss Surg 9(2): 76-81.
 INTRODUCTION: In children, the choice between percutaneous pinning (PP) and open pinning fixation (OPF) for the surgical treatment of fractures of the distal humerus remains controversial, especially the PP method for internal humeral condylar (IHC) fractures. PATIENTS AND METHODS: Eighty fractures of the distal humerus in children were treated surgically in our hospital over a ten year period. 47% (n = 38) were supracondylar (SC), 20% (n = 16) comminuted (COM), 18% (n = 14) internal humeral condylar (IHC), and 15% (n = 12) lateral humeral condylar (LHC). We used PP, OPF and three times osteosynthesis with screws. RESULTS: In comparison to OPF, PP reduced the length of hospitalization in SC fractures (2.8 versus 6.1 days) and IHC fractures (2.4 versus five days). It reduced the risk of extension deficiency (11.1% versus 15%) and of cubitus valgus (0% versus 20%) in SC fractures, and of cubitus varus in IHC fractures (0% versus 11.1%). However it induced a higher rate of cubitus valgus (11.1% versus 20%) in IHC fractures, one persistent neurological motor deficiency (radial nerve) and four cases of transitional neurological involvement (ulnar nerve). CONCLUSIONS: PP is a good surgical method for SC and for also for IHC fractures, if performed by experienced surgeons so as to avoid neurological damage.

De Jonge, R. R., I. N. Van Schaik, et al. (2003). "Expression of complement components in the peripheral nervous system." Hum Mol Genet.
 We have generated a SAGE (serial analysis of gene expression) library of normal sciatic nerve and found tags encoding for mRNAs of the complement system highly represented. RNA (RT-PCR and Northern blot hybridisation) and protein (Western blot analysis and immunohistochemistry) studies confirmed these findings. High expression of classical pathway components, alternative pathway components and inhibitory components was observed in specific regions of the sciatic nerve. The first components of complement were found in axons, whereas the inhibitory components were detected in the perineurium, thereby protecting the nerve from a complement attack. Immunoreactivity towards activated complement factors was noted in post traumatic neuromas and after acute crush injury, which exemplify nerve regeneration and degeneration. We propose that local production of complement in the peripheral nervous system participates in the protection of healthy nerve and is needed for efficient clearance of myelin after injury: a prerequisite for normal regeneration and remyelination of the peripheral nerve.

Deda, G., H. Caksen, et al. (2003). "A case of Marin-Amat syndrome." Genet Couns 14(1): 115-7.
 
Degidi, M., G. Petrone, et al. (2003). "Bone contact around acid-etched implants: a histological and histomorphometrical evaluation of two human-retrieved implants." J Oral Implantol 29(1): 13-8.
 The surface characteristics of dental implants play an important role in their clinical success. One of the most important surface characteristics of implants is their surface topography or roughness. Many techniques for preparing dental implant surfaces are in clinical use: turning, plasma spraying, coating, abrasive blasting, acid etching, and electropolishing. The Osseotite surface is prepared by a process of thermal dual etching with hydrochloric and sulfuric acid, which results in a clean, highly detailed surface texture devoid of entrapped foreign material and impurities. This seems to enhance fibrin attachment to the implant surface during the clotting process. The authors retrieved 2 Osseotite implants after 6 months to repair damage to the inferior alveolar nerve. Histologically, both implants appeared to be surrounded by newly formed bone. No gaps or fibrous tissues were present at the interface. The mean bone-implant contact percentage was 61.3% (+/- 3.8%).

Deininger, S. O., L. Rajendran, et al. (2003). "Identification of teleost Thy-1 and association with the microdomain/lipid raft reggie proteins in regenerating CNS axons." Mol Cell Neurosci 22(4): 544-54.
 During regeneration, retinal ganglion cell axons in fish upregulate a cell surface protein that is recognized by the monoclonal antibody (mAB) M802. M802 antigen appeared to be linked to the intracellular, membrane-associated lipid raft/microdomain proteins reggie-1 and reggie-2 that were previously shown to be reexpressed in axon-regenerating neurons [Development 124 (1997), 577]. Here, we report the isolation of the M802 antigen and its identification as the teleost homolog of mammalian Thy-1. Fish Thy-1 is detected in the same detergent-insoluble lipid raft fractions from a fibroblast cell line and from axon regenerating retinae as reggie-1 and 2. Importantly, mAB M802 coimmunoprecipitates reggie-1 and 2 from this lipid raft fraction, implying that fish Thy-1 and reggies interact. This correlates with their colocalization in growing cell processes after M802 antigen/Thy-1 activation with mAB M802. These findings suggest a role of clustered M802 antigen/Thy-1 in reggie raft microdomains for cell growth and axon regeneration.

DeLucia, T. A., J. J. Conners, et al. (2003). "Use of a cell line to investigate olfactory ensheathing cell-enhanced axonal regeneration." Anat Rec 271B(1): 61-70.
 Olfactory ensheathing cells (OECs), a unique type of macroglia required for normal olfactory axonal regeneration throughout the lifetime of an individual, have been shown to have regeneration-enhancing properties when used to treat various neuronal injuries. Availability of OECs is a hurdle facing future clinical use of the cells for spinal cord injury (SCI) therapy. The number of OECs that can realistically be harvested from each animal is limited, and ensuring a pure cell population is difficult. We have begun to characterize a nonsyngeneic strain of OECs, i.e., from a homogenous OEC clonal cell line (nOECs). The purpose of this study was to determine whether nOECs have the same properties and provide the same functional recovery after SCI, as primary cultures of OECs. The results indicate that nOECs survive in vivo, produce growth-promoting proteins, and possess regeneration-promoting capabilities. Spinal cord injured rats that were treated with nOECs performed significantly better on functional tests than injured control animals beginning at 5 weeks after operation. In summary, evidence of nOEC regeneration-promoting capabilities suggests that this cell line can be used as potential therapy in SCI research.

Dervan, A. G. and B. L. Roberts (2003). "Reaction of spinal cord central canal cells to cord transection and their contribution to cord regeneration." J Comp Neurol 458(3): 293-306.
 After transection, the spinal cord of the eel Anguilla quickly regrows and reconnects, and function recovers. We describe here the changes in the central canal region that accompany this regeneration by using serial semithin plastic sections and immunohistochemistry. The progress of axonal regrowth was followed in material labeled with DiI. The canal of the uninjured cord is surrounded by four cell types: S-100-immunopositive ependymocytes, S-100- and glial fibrillary acidic protein (GFAP)-immunopositive tanycytes, vimentin-immunopositive dorsally located cells, and lateral and ventral liquor-contacting neurons, which label for either gamma-aminobutyric acid (GABA) or tyrosine hydroxylase (TH). After cord transection, a new central canal forms rapidly as small groups of cells at the leading edges of the transection create flat "plates" that serve as templates for subsequent formation of the lateral and dorsal walls. Profile counts and 5-bromo-2'-deoxyuridine immunohistochemistry indicate that these cells are dividing rapidly during the first 20 days of the repair process. The newly formed canal, which bridges the transection by day 10 but is not complete until about day 20, is greatly enlarged (</=100 times) and is dominated by ependymocytes that are vimentin immunopositive, but cells expressing GABA, TH, and GFAP do not appear until days 11, 13, and 16, respectively. The proliferating ependyma do not provide a supportive scaffold for the regrowing axons, inasmuch as some have crossed the bridge before the canal has formed. However, their modified phenotype suggests a role, possibly trophic, for the central canal region following injury.

Desbarats, J., R. B. Birge, et al. (2003). "Fas engagement induces neurite growth through ERK activation and p35 upregulation." Nat Cell Biol 5(2): 118-25.
 Fas (also known as CD95), a member of the tumour-necrosis receptor factor family of 'death receptors', can induce apoptosis or, conversely, can deliver growth stimulatory signals. Here we report that crosslinking Fas on primary sensory neurons induces neurite growth through sustained activation of the extracellular-signal regulated kinase (ERK) pathway and the consequent upregulation of p35, a mediator of neurite outgrowth. In addition, functional recovery after sciatic nerve injury is delayed in Fas-deficient lpr mice and accelerated by local administration of antibodies against Fas, which indicates that Fas engagement may contribute to nerve regeneration in vivo. Our findings define a role for Fas as an inducer of both neurite growth in vitro and accelerated recovery after nerve injury in vivo.

Dieringer, N. (2003). "Activity-Related Postlesional Vestibular Reorganization." Ann N Y Acad Sci 1004: 50-60.
 The synaptic convergence patterns of semicircular canal and macular afferent nerve inputs onto second-order vestibular neurons reorganize in adult frogs after a change in the activity of vestibular nerve afferent fibers. Axotomized afferent nerve fibers become silent after a vestibular nerve lesion, and second-order vestibular target neurons become disfacilitated. These changes initiate an activity-related process that was studied in detail in vitro two months after a section of the ramus anterior (RA) of N. VIII. The postlesional reaction results in an expansion of signals, preferentially from intact, remaining afferent nerve fibers, but also from excitatory commissural and spinal ascending fibers. This process of expansion takes weeks, is graded in its extent, and reversible in case of a nerve regeneration, but is not competitive, i.e., the synaptic contacts from axotomized afferent nerve fibers are maintained without a change in their efficacy. Postlesional synaptic reorganization in the brainstem is restricted to the operated side, underlies the improved responsiveness of disfacilitated second-order vestibular neurons, but also their altered spatial response tuning. The functional consequences of this reorganization were studied in vivo two months after RA nerve section by recording abducens nerve responses during linear or angular accelerations. The vector orientations of best responses of the abducens nerve of chronic RA frogs evoked by linear or angular acceleration differed from the vector orientations of controls. In chronic RA frogs, linear acceleration evoked contralesional abducens nerve responses that originated from the utricle on the intact side and from the lagena, a vertical macular organ in frogs. Such an inadequate lagenar response component was absent in controls and in the ipsi-lesional abducens nerve of chronic RA frogs. Similar differences were detected in the direction of abducens nerve responses of chronic RA frogs during angular acceleration. Thus, compensatory vestibulo-ocular reflexes of chronic RA frogs became more symmetric in gain, but less precise in direction.

Diiulio, R. (2003). "The new alternatives." Rehab Manag 16(6): 38-41, 50.
 
Dolbeare, D. and J. D. Houle (2003). "Restriction of axonal retraction and promotion of axonal regeneration by chronically injured neurons after intraspinal treatment with glial cell line-derived neurotrophic factor (GDNF)." J Neurotrauma 20(11): 1251-61.
 The response of supraspinal neurons to acute or delayed treatment with GDNF following a spinal cord injury was examined. A cervical level 3 hemisection lesion cavity was created by tissue aspiration in adult, female rats. In one experiment gel foam saturated with GDNF was placed into the lesion cavity immediately after injury to determine if the extent of axonal retraction was affected by neurotrophic factor treatment. One week prior to sacrifice animals received a microinjection of biotinylated dextran amine (BDA) into the red nucleus and reticular formation to label descending spinal pathways by anterograde transport mechanisms. Animals were sacrificed 1 or 4 weeks after injury and treatment with GDNF. The terminal end of injured BDA-labeled rubrospinal and reticulospinal tract axons was identified and the distance from the lesion was measured. In comparison to PBS-treated animals, GDNF-treatment resulted in a significant decrease in the extent of axonal retraction of both rubrospinal and reticulospinal tract axons at 1 week after spinal cord injury for both tracts. At 4 weeks after injury the mean distance from the lesion was less than 240 microm following GDNF-treatment for both tracts, compared to over 480 microm following PBS-treatment. In the second experiment injured supraspinal neurons were labeled by retrograde transport of True Blue that had been placed into the lesion cavity. One month later scar tissue was removed from the cavity by aspiration to enlarge the cavity by approximately 500 microm in a rostral direction. GDNF-saturated gel foam was placed into the cavity for 60 min prior to apposition of an autologous peripheral nerve (PN) graft to the rostral cavity wall. One month later Nuclear Yellow was applied to the distal end of the PN graft and animals were sacrificed after 2 days. The number of supraspinal neurons containing both True Blue and Nuclear Yellow was counted as a measure of axonal regeneration by chronically injured neurons. There was a seven-fold increase in the number of regenerating neurons after GDNF-treatment, with the majority (65%) of dual-labeled neurons located within the reticular formation. These results indicate that GDNF has neuroprotective effects when provided acutely after injury and promotes axonal regeneration when provided in a chronic injury situation.

Donnerer, J. (2003). "Regeneration of primary sensory neurons." Pharmacology 67(4): 169-81.
 Primary sensory neurons have an inherent capacity for regeneration of their cut, crushed, or chemically lesioned axons. This capacity is displayed to a much greater extent after lesions of the peripheral axons than after lesions of their centrally directed axons. Additionally, the surrounding tissue determines to a significant extent the degree of recovery: whereas the peripheral nerve tissue provides neurotrophic support and a favorable environment for axonal growth, the central terminals of primary sensory neurons face a non-permissive and inhibitory glial tissue. Mechanical lesions of the peripheral axons of dorsal root ganglion (DRG) sensory neurons can be repaired by the intrinsic regenerative capacity of the neuron itself, when outgrowing axons from the proximal stump are able to transverse the tissue scar and reach the distal stump of the nerve. Bridging the gap with an autologous nerve graft or a short artificial graft filled with nerve growth factor (NGF) can improve recovery. Neurotoxic lesions of the axon terminals are effectively recovered by intermittent local or systemic NGF injections. A recovery from a diabetic sensory neuropathy probably requires the continuous delivery of NGF or additional neurotrophic factors. A recovery from a dorsal rhizotomy or from a dorsal column lesion can possibly be achieved by the concomitant transgene-mediated overexpression of neurotrophins, the transformation of the DRG neuron cells to a competence for regrowth, and the counteraction of the growth-inhibitory nature of the central nervous system tissue.

Duan, Y., S. J. Haugabook, et al. (2003). "Methylene blue blocks cGMP production and disrupts directed migration of microglia to nerve lesions in the leech CNS." J Neurobiol 57(2): 183-92.
 Migration and accumulation of microglial cells at sites of injury are important for nerve repair. Recent studies on the leech central nervous system (CNS), in which synapse regeneration is successful, have shown that nitric oxide (NO) generated immediately after injury by endothelial nitric oxide synthase (eNOS) stops migrating microglia at the lesion. The present study obtained results indicating that NO may act earlier, on microglia migration, and aimed to determine mechanisms underlying NO's effects. Injury induced cGMP immunoreactivity at the lesion in a pattern similar to that of eNOS activity, immunoreactivity, and microglial cell accumulation, which were all focused there. The soluble guanylate cyclase (sGC) inhibitor methylene blue (MB) at 60 microM abolished cGMP immunoreactivity at lesions and blocked microglial cell migration and accumulation without interfering with axon conduction. Time-lapse video microscopy of microglia in living nerve cords showed MB did not reduce cell movement but reduced directed movement, with significantly more cells moving away from the lesion or reversing direction and fewer cells moving toward the lesion. The results indicate a new role for NO, directing the microglial cell migration as well as stopping it, and show that NO's action may be mediated by cGMP.

Dubreuil, C. I., M. J. Winton, et al. (2003). "Rho activation patterns after spinal cord injury and the role of activated Rho in apoptosis in the central nervous system." J Cell Biol 162(2): 233-43.
 Growth inhibitory proteins in the central nervous system (CNS) block axon growth and regeneration by signaling to Rho, an intracellular GTPase. It is not known how CNS trauma affects the expression and activation of RhoA. Here we detect GTP-bound RhoA in spinal cord homogenates and report that spinal cord injury (SCI) in both rats and mice activates RhoA over 10-fold in the absence of changes in RhoA expression. In situ Rho-GTP detection revealed that both neurons and glial cells showed Rho activation at SCI lesion sites. Application of a Rho antagonist (C3-05) reversed Rho activation and reduced the number of TUNEL-labeled cells by approximately 50% in both injured mouse and rat, showing a role for activated Rho in cell death after CNS injury. Next, we examined the role of the p75 neurotrophin receptor (p75NTR) in Rho signaling. After SCI, an up-regulation of p75NTR was detected by Western blot and observed in both neurons and glia. Treatment with C3-05 blocked the increase in p75NTR expression. Experiments with p75NTR-null mutant mice showed that immediate Rho activation after SCI is p75NTR dependent. Our results indicate that blocking overactivation of Rho after SCI protects cells from p75NTR-dependent apoptosis.

Dunlop, S. A. (2003). "Axonal sprouting in the optic nerve is not a prerequisite for successful regeneration." J Comp Neurol 465(3): 319-34.
 Axonal sprouting, the production of axons additional to the parent one, occurs during optic nerve regeneration in goldfish and the frog Rana pipiens, with numbers of regenerate axons exceeding normal values four- to sixfold (Murray [1982] J. Comp. Neurol. 209:352-362; Stelzner and Strauss [1986] J. Comp. Neurol. 245:83-103). To determine whether axonal sprouting is a prerequisite for regeneration, the frog Litoria moorei was examined, a species that undergoes successful optic nerve regeneration but with a different time course compared with R. pipiens. Sprouting was assessed, as in goldfish and R. pipiens, from electron microscopic counts between the lesion and chiasm. However, disconnected axons that persist after axotomy would have falsely elevated the counts. The suspected overlap of these two axon populations was confirmed by labeling regenerate axons anterogradely with DiI (1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate) and disconnected ones retrogradely with DiA (4-4-dihexadecylaminostyrl 1-N methylpyridinium iodide). Numbers of disconnected axons were estimated after preventing regeneration and subtracted from numbers in regenerate nerves. Throughout, the total number of regenerate axons was approximately one third lower than normal (P < 0.05) supporting a previous finding of minimal axonal sprouting in L. moorei (Dunlop et al. [2002] J. Comp. Neurol. 446:276-287). The validity of the subtractive electron microscopic method was confirmed by retrograde labeling to estimate numbers of retinal ganglion cells whose axons had crossed the lesion; values were approximately one third lower than normal. The data suggest that sprouting is not essential for either axon outgrowth or topographic map refinement.

Duprey-Diaz, M. V., J. M. Blagburn, et al. (2003). "Neurotrophin-3 and TrkC in the frog visual system: changes after axotomy." Brain Res 982(1): 54-63.
 Neurotrophins are potent regulators of the survival of different neuronal populations in the CNS. Little is known of the immunodistribution of neurotrophin-3 (NT-3) and tyrosine kinase C (TrkC) receptor in the frog visual system, which can successfully regenerate and recover vision after injury. In this study we show that both NT-3 and TrkC are present in the frog retina and tectum, and that their distribution changes after optic nerve transection. Both NT-3 and TrkC are present in the ganglion cell layer, inner nuclear layer, nerve fiber layer and outer plexiform layer, and in Muller cells of control retinas. Quantification of identified RGCs shows that there are only small changes in the proportion, or intensity, of NT-3 immunostained cells surviving after axotomy and regeneration. Muller cell staining, however, is increased. TrkC staining in the retina does not change after axotomy. In the tectum, NT-3 immunoreactivity is present in the retinorecipient layer 9, and in radial processes of neurons and ependymoglia. TrkC is present in ependymoglia and in tectal neurons. After axotomy or colchicine treatment fewer NT-3-immunoreactive processes are present in layer 9 and there is decreased staining of tectal neurons. These data are consistent with the hypothesis that NT-3 is synthesized in the retina and anterogradely transported to the tectum. TrkC immunostaining, on the other hand, increases in tectal cells after optic nerve transection, suggesting that it may be regulated by the supply of NT-3 from the retina.

Echeverri, K. and E. M. Tanaka (2003). "Electroporation as a tool to study in vivo spinal cord regeneration." Dev Dyn 226(2): 418-25.
 Tailed amphibians such as axolotls and newts have the unique ability to fully regenerate a functional spinal cord throughout life. Where the cells come from and how they form the new structure is still poorly understood. Here, we describe the development of a technique that allows the visualization of cells in the living animal during spinal cord regeneration. A microelectrode needle is inserted into the lumen of the spinal cord and short rapid pulses are applied to transfer the plasmids encoding the green or red fluorescent proteins into ependymal cells close to the plane of amputation. The use of small, transparent axolotls permits imaging with epifluorescence and differential interference contrast microscopy to track the transfected cells as they contribute to the spinal cord. This technique promises to be useful in understanding how neural progenitors are recruited to the regenerating spinal cord and opens up the possibility of testing gene function during this process.

Edstrom, A. and P. A. Ekstrom (2003). "Role of phosphatidylinositol 3-kinase in neuronal survival and axonal outgrowth of adult mouse dorsal root ganglia explants." J Neurosci Res 74(5): 726-35.
 Adult ganglionic peripheral neurons have lost dependence on target-derived neurotrophin signaling for survival and regeneration after injury. To understand the mechanisms required to sustain such processes at maturity, we are studying neuronal survival and axonal outgrowth of adult mouse dorsal root ganglia (DRG) explants. We have here examined the role of phosphatidylinositol 3-kinase (PI3-K) activity. Both neuronal survival and axonal outgrowth of spontaneously growing preparations were decreased significantly by the PI3-K inhibitor LY294002 as was the increased outgrowth caused by nerve growth factor or glial cell line-derived factor. Inhibition of PI3-K activity promoted neuronal cell death to the same extent in the presence as in the absence of a growth factor, whereas inhibition of mitogen-activated protein kinase, MAPK, lacked effect. Using a compartmentalized system, it could be shown that only axonal outgrowth was decreased when the outgrowth region only was exposed to LY294002. Already-formed growth cones showed morphological changes within 5-10 min after exposure to LY294002. Akt (PKB) is one downstream effector of PI3-K. Immunofluorescence revealed the presence of activated Akt in DRG cell bodies and in axonal growth cones. Immunoreactivity was decreased by PI3-K inhibition. The results suggest that Akt is constitutively active in adult DRG neurons, and that PI3-K mediated processes are involved in neuronal survival of one or more DRG neuronal subpopulations and also in axonal elongation. The possible significance of Akt signaling for these effects is discussed.

Efimov, I. R. (2003). "Fibrillation or neurillation: back to the future in our concepts of sudden cardiac death?" Circ Res 92(10): 1062-4.
 
Eguchi, Y., M. Ogiue-Ikeda, et al. (2003). "Control of orientation of rat Schwann cells using an 8-T static magnetic field." Neurosci Lett 351(2): 130-2.
 Schwann cells aid in neuronal regeneration in the peripheral nervous system via guiding the regrowth of axons. In this study, we investigated the magnetic orientation of Schwann cells, and of a mixture of Schwann cells and collagen, after an 8-tesla magnetic field exposure. We obtained cultured Schwann cells from dissected sciatic nerves of neonatal rats. After 60 h of magnetic field exposure, Schwann cells oriented parallel to the magnetic fields. In contrast, the mixture of Schwann cells and collagen, Schwann cells oriented in the direction perpendicular to the magnetic field after 2 h of magnetic field exposure. In this case, Schwann cells aligned along the collagen fiber oriented by magnetic fields. The magnetic control of Schwann cell alignment is useful in medical engineering applications such as nerve regeneration.

Ekstrom, P. A., U. Mayer, et al. (2003). "Involvement of alpha7beta1 integrin in the conditioning-lesion effect on sensory axon regeneration." Mol Cell Neurosci 22(3): 383-95.
 Conditioning lesions of peripheral nerves improve axonal regeneration after injury and involve changes in expression of proteins required for axonal growth. Integrin alpha7beta1 expression in motor and sensory neurons increases following nerve lesions and motor axon regeneration is impaired in alpha7 integrin KO mice (J. Neurosci. 20, 1822-1830). To investigate the role of alpha7beta1 integrin in sensory axon regeneration, dorsal root ganglia of adult mice were cultured in gels of laminin-rich extracellular matrix (Matrigel) or collagen. Normal dorsal root ganglia in Matrigel or collagen supplemented with laminin showed spontaneous axonal outgrowth, which was greatly increased in conditioned preparations, but only in the presence of laminin. Conditioned dorsal root ganglia from normal mice cultured with a blocking antibody to beta1 integrin and from alpha7 integrin KO mice showed reduced axonal growth in both Matrigel- and laminin-supplemented collagen gels. Enhanced axonal regeneration after conditioning lesions therefore involves increased responsiveness to laminin and integrin alpha7beta1 expression.

El-Gammal, T. A., A. El-Sayed, et al. (2003). "Surgical treatment of brachial plexus traction injuries in children, excluding obstetric palsy." Microsurgery 23(1): 14-7.
 Traumatic brachial plexus injuries in children, excluding birth palsy, are seldom reported. In this study, we report on 11 cases operated upon between 1995-1998, and followed for at least 30 months. All patients were males with an average age of 11 years (range, 3-16 years). The denervation time averaged 3.8 months (range, 1-8 months). Eight patients had two or more root avulsions; two had additional severe infraclavicular injuries. In total, 6 grafting and 25 extraplexal neurotization procedures were used. Donor nerves included the intercostal nerves, phrenic nerve, spinal accessory nerve, and contralateral C7 root. Elbow flexion was restored in all but 2 cases. Shoulder abduction varied from 30-90 degrees, according to the method of reconstruction. Triceps recovered in 2 cases and finger and wrist extensors in 1 case. Wrist and finger flexion was obtained in 1 case. Sensory recovery in the palm reached S2/S2+. Harvesting the phrenic nerve and the contralateral C7 root resulted in no residual morbidity. Compared to adults, children have a higher incidence of root avulsion, no deafferentiation pain, a higher incidence of associated skeletal injuries, and the same recovery rate of elbow and shoulder functions following plexus reconstruction, but recovery is faster. Given the frequency of root avulsions, neurotization is often required.

Ellezam, B., J. Bertrand, et al. (2003). "Vaccination stimulates retinal ganglion cell regeneration in the adult optic nerve." Neurobiol Dis 12(1): 1-10.
 We examined whether vaccination of adult rats with spinal cord homogenate (SCH) can promote regeneration of retinal ganglion cells (RGCs) after microcrush lesion of the optic nerve. Injured animals vaccinated with SCH showed axon growth into the optic nerve and such regeneration was not observed in animals vaccinated with liver homogenate (LH). Regeneration was not a consequence of neuroprotection since our vaccine did not protect RGCs from axotomy-induced cell death. Sera of vaccinated animals were tested for antibodies against myelin-associated glycoprotein, NogoA, Nogo-66 receptor, or chondroitin sulphate proteoglycans (CSPG), but no significant levels were detected. Antibodies to myelin basic protein were present in the serum of some SCH-vaccinated animals. In culture, serum from SCH-vaccinated animals promoted RGC growth on myelin but not on CSPG. Our results show that the effect of the pro-regenerative vaccine is mediated by antibodies to SCH. However, we were not able to detect a significant immune reaction to growth inhibitory proteins, suggesting alternative mechanisms for the success of vaccination to promote regeneration.

Ellis, R. A., M. J. Brenner, et al. (2003). "Use of mixed lymphocyte reaction to identify subimmunosuppressive FK-506 levels in mice." Microsurgery 23(3): 276-82.
 The immunosuppressive agent FK-506 has a well-described neuroregenerative effect that is mediated by a mechanism independent of calcineurin inhibition. FK-506 levels that fall below the threshold for immunosuppression could therefore potentially enhance nerve regeneration while minimizing toxicity. The purpose of this study was to characterize the dose-dependent effects of FK506 on T-cell proliferation, and establish a subimmunosuppressive dosing regimen for FK-506 in mice. Forty BALB/cJ mice were randomized to four groups corresponding to 0, 0.25, 0.5, or 1.0 mg/kg/day doses of FK-506. Ten days postoperatively, animals were sacrificed, and mixed lymphocyte reaction assays were performed to quantify the immune response to nerve allografts. Mice receiving 0.25 and 0.5 mg/kg/day of FK-506 exhibited a robust T-cell proliferation response, with stimulation indices approaching those of untreated animals. Mice treated with 1.0 mg/kg/day of FK-506 demonstrated significantly decreased T-cell proliferation. These results establish 0.5 mg/kg/day as an upper limit for subimmunosuppressive FK-506 administration.

Elmas, C., D. Erdogan, et al. (2003). "Expression of growth factors in fetal human olfactory mucosa during development." Growth Dev Aging 67(1): 11-25.
 The olfactory epithelium is one of the unique sites in the mammalian nervous system at which there is continual neurogenesis. Constant turnover of primary sensory neurons in the periphery results in remodeling of neuronal circuits and synapses in the olfactory bulb throughout life. Most of the specific mechanisms and factors that control this process are still unknown. Recent studies suggest that growth factors and their receptors may play a significant role in the development and continuous regeneration of olfactory neurons. In this study, standard immunohistochemical methods were used to detect the presence of epidermal growth factor-receptor, transforming growth factor-alpha and nerve growth factor-beta in the human olfactory epithelium of different developmental stages (7 to 30 weeks) in formalin-fixed tissue specimens. Immunoreactivity to epidermal growth factor-receptor was seen in basal cells, supporting cells, neurons, apical cytoplasmic region of olfactory epithelium, lamina propria and gland's cell cytoplasm. Similarly transforming growth factor-alpha immunoreactivity was recognized in basal cells, supporting cells, apical cytoplasmic region of olfactory epithelium, lamina propria and gland's cell cytoplasm but not in neurons. Nerve growth factor-beta detection was restricted on olfactory nerve cells, dentritic knob and basal cell layer. So, this study confirms the development of human olfactory mucosa and shows epidermal growth factor-receptor, transforming growth factor-alpha and nerve growth factor-beta proteins are reliable markers for developing olfactory epithelium.

Emerick, A. J., E. J. Neafsey, et al. (2003). "Functional reorganization of the motor cortex in adult rats after cortical lesion and treatment with monoclonal antibody IN-1." J Neurosci 23(12): 4826-30.
 We previously reported anatomical plasticity in the adult motor cortex after a unilateral sensorimotor cortex (SMC) lesion and treatment with monoclonal antibody (mAb) IN-1, which permits neurite outgrowth from the intact, opposite cortex into deafferented subcortical targets. This study was designed to investigate whether treatment with the mAb IN-1 after SMC lesion in the adult leads to functional reorganization of the intact, opposite motor cortex. Adult rats underwent unilateral SMC aspiration lesion and treatment with either mAb IN-1 or control antibody, or no treatment. After a 6 week survival period, the intact, opposite forelimb motor cortex was explored using intracortical microstimulation to evoke forelimb movements. A dramatic increase in ipsilateral movements of the lesion-impaired forelimb was found in animals treated with mAb IN-1 compared with control animals. These results resembled our previous findings of cortical reorganization in the spared hemisphere after neonatal cortical lesion and without any additional treatment. These results show that, after adult cortical lesion, treatment with mAb IN-1 induces a functional reorganization of the intact, opposite motor cortex.

Emanueli, C., P. Schratzberger, et al. (2003). "Paracrine control of vascularization and neurogenesis by neurotrophins." Br J Pharmacol 140(4): 614-9.
 The neuronal system plays a fundamental role in the maturation of primitive embryonic vascular network by providing a paracrine template for blood vessel branching and arterial differentiation. Furthermore, postnatal vascular and neural regeneration cooperate in the healing of damaged tissue. Neurogenesis continues in adulthood although confined to specific brain regions. Following ischaemic insult, neural staminal cells contribute towards the healing process through the stimulation of neurogenesis and vasculogenesis. Evidence indicates that nerves and blood vessels exert a reciprocal control of their own growth by paracrine mechanisms. For instance, guidance factors, including vascular endothelial growth factor A (VEGF-A) and semaphorins, which share the ability of binding neuropilin receptors, play a pivotal role in the tridimensional growth pattern of arterial vessels and nerves. Animal models and clinical studies have demonstrated a role of VEGF-A in the pathogenesis of ischaemic and diabetic neuropathies. Further, supplementation with VEGF-A ameliorates neuronal recovery by exerting protective effects on nerves and stimulating reparative neovascularization. Human tissue kallikrein, a recently discovered angiogenic and arteriogenic factor, accelerates neuronal recovery by stimulating the growth of vasa nervorum. Conversely, the neurotrophin nerve growth factor, known to regulate neuronal survival and differentiation, is now regarded as a stimulator of angiogenesis and arteriogenesis. These results indicate that angiogenesis and neurogenesis are paracrinally regulated by growth factors released by endothelial cells and neurons. Supplementation of these growth factors, alone or in combination, could benefit the treatment of ischaemic diseases and neuropathies.

Emes, R. D., W. Z. Wang, et al. (2003). "HmCRIP, a cysteine-rich intestinal protein, is expressed by an identified regenerating nerve cell." FEBS Lett 533(1-3): 124-8.
 A Hirudo medicinalis cDNA isolated from regenerating CNS tissue at 24 h post-axotomy was identified as a leech homologue of the mammalian cysteine-rich intestinal proteins (CRIPs) and named HmCRIP. HmCRIP is up-regulated within 6 h of axotomy, peaking at 24 h. This is the first demonstration of a CRIP homologue in regenerating CNS and in a serotonergic neurone. In rodents CRIP is an important factor in the regulation of the inflammatory immune response through control of Th1/Th2 differentiation. The role of HmCRIP in the regeneration competent environment of the annelid central nervous system is discussed.

Erie, J. C., S. V. Patel, et al. (2003). "Aberrant corneal nerve regeneration after PRK." Cornea 22(7): 684-6.
 PURPOSE: To report a case of aberrant corneal nerve regeneration after myopic photorefractive keratectomy (PRK). METHODS: One patient underwent bilateral PRK to correct a refractive error of -5.50 D in each eye. Thirteen months after the original PRK, the left eye underwent an uncomplicated PRK reoperation to correct a regression of -1.00 D. The central corneas were examined by confocal microscopy preoperatively in both eyes, at 1 and 2 years after the original PRK in the right eye, and before and 1 and 2 years after the PRK reoperation in the left eye. RESULTS: Aberrant anterior stromal nerves with a coiled course and irregular branching pattern were identified 22 micro m deep to the most anterior keratocyte layer at 1 year after the PRK reoperation in the left eye and remained unchanged 2 years after reoperation. No abnormal stromal nerves were identified in the left eye before the reoperation or at any time in the right eye. CONCLUSION: Aberrant regeneration of corneal stromal nerves may occur after myopic PRK reoperation.

Evans, G. R. and K. Brandt (2003). "Peripheral nerve regeneration: the effects of postoperative irradiation." Plast Reconstr Surg 111(6): 2023-4.
 
Evers, B. M., I. L. Weissman, et al. (2003). "Stem cells in clinical practice." J Am Coll Surg 197(3): 458-78.
 
Eyupoglu, I. Y., I. Bechmann, et al. (2003). "Modification of microglia function protects from lesion-induced neuronal alterations and promotes sprouting in the hippocampus." Faseb J 17(9): 1110-1.
 Primary neuronal destruction in the central nervous system triggers rapid changes in glial morphology and function, after which activated glial cells contribute to secondary neuronal changes. Here we show that, after entorhinal cortex lesion, activation of microglia, but not other glial cells, leads to massive secondary dendritic changes of deafferentiated hippocampal neurons. Blocking of microglial activation in vivo reduced this secondary neuronal damage and enhanced regenerative axonal sprouting. In contrast, abolishing astrocytes or oligodendroglia did not result in specific neuronal changes. Furthermore, primary damage leads to an interleukin 1beta up-regulation, which is attenuated by the immuno-modulator transforming growth factor beta1, whereas tumor necrosis factor alpha is not affected. Modification of microglial activity following denervation of the hippocampus protects neurons from secondary dendritic alterations and therefore enables their reinnervation. These data render activated microglia a putative therapeutic target during the course of axonal degeneration.

Faivre, S., A. Lim, et al. (2003). "Adjacent and spontaneous neurotization after distal digital replantation in children." Plast Reconstr Surg 111(1): 159-65; discussion 166.
 In an exclusively pediatric population, this retrospective study examined the functional and aesthetic results after distal replantation without nerve suture. The aim was to demonstrate, in the child, the presence of spontaneous nervous regeneration resulting in a fingertip pulp with discriminatory sensation. Eight amputations in eight children with a mean age of 9 years and 2 months on the day of the accident were reviewed. The cases were managed by a single surgeon over a period of 8 years and were collected from two different hand centers. The patients were then examined by a different surgeon, and the data were collected. Sensibility was evaluated using the Weber, Semmes-Weinstein, and wrinkle tests. The results were excellent, with mean values of 4.6 mm for the Weber test, 3.3 for the Semmes-Weinstein test, and a positive wrinkle test in all subjects. All patients thus recovered discriminatory sensation with minimal aesthetic sequelae. The usual factors adversely affecting the results of the replantation (ischemic time, level and mechanism of the amputation, and quality of the venous return) were examined, but no statistical analysis was performed because of the small sample size. This study demonstrates the presence of the clinical phenomenon of adjacent neurotization in the absence of nerve repair. It thus confirms that children are excellent candidates for replantation of the distal extremities, even when nerve suture is not performed.

Fansa, H. and G. Keilhoff (2003). "[Factors influencing nerve regeneration]." Handchir Mikrochir Plast Chir 35(2): 72-82.
 This paper describes the most important cellular and molecular factors that influence nerve regeneration. The first prerequisite for axonal regeneration is survival of the neuron. This depends on neuron type, age, and the degree and proximity of the injury to the cell body. Spinal motoneurons are less susceptible to injury-induced death than cranial motoneurons and sensory neurons. The surviving neurons undergo changes characteristic of a switch from a transmitting mode to a growing mode. They produce various neurotrophic factors and their receptors influencing the neuron and the non-neuronal cells such as Schwann cells. The distal nerve stump undergoes degenerative processes including removal of axons and phagocytosis of myelin debris, the so-called Wallerian degeneration. Until the second day phagocytosis is done by Schwann cells, hematogenous macrophages invade the distal stump at the second day and phagocyte the whole debris within two weeks. Devoid of axonal contact, the myelinating Schwann cells switch their function from myelination to growth support for the regenerating axons, including cell proliferation, downregulation of myelin components and upregulation of neurotrophic factors. Additionally, the Schwann cells form the so-called Bands of Bungner, cell columns serving as pathway for the growing axon. Trophic factors, cell adhesion molecules and extracellular matrix influence the neuron, the growing axon and the endorgan as well as the non-neuronal cells such as Schwann cells, fibroblasts and macrophages. Application of drugs or trophic substances to enhance nerve regeneration after trauma and reconstruction is in the very beginning, and thus requires further experimental and clinical studies. Experimentally, FK 506 was found to support axonal regeneration after crush lesions and nerve grafting. Growth factors are currently administered clinically in other neurological diseases.

Fansa, H., T. Dodic, et al. (2003). "Tissue engineering of peripheral nerves: Epineurial grafts with application of cultured Schwann cells." Microsurgery 23(1): 72-7.
 After a simple nerve lesion, primary microsurgical suture is the treatment of choice. A nerve gap has to be bridged, with a nerve graft sacrificing a functioning nerve. Alternatively, tissue engineering of nerve grafts has become a subject of experimental research. It is evident that nerve regeneration requires not only an autologous, allogenous, or biodegradable scaffold, but additional interactions with regeneration-promoting Schwann cells. In this study, we compared epineurial and acellularized epineurial tubes with and without application of cultured Schwann cells as alternative grafts in a rat sciatic nerve model. Autologous nerve grafts served as controls. Evaluation was performed after 6 weeks; afterwards, sections of the graft and distal nerve were harvested for histological and morphometrical analysis. Compared to controls, all groups showed a significantly lower number of axons, less well-shaped remyelinizated axons, and a delay in clinical recovery (e.g., toe spread). The presented technique with application of Schwann cells into epineurial tubes did not offer any major advantages for nerve regeneration. Thus, in this applied model, neither the implantation of untreated nor the implantation of acellularized epineurial tubes with cultured Schwann cells to bridge nerve defects was capable of presenting a serious alternative to the present gold standard of conventional nerve grafts for bridging nerve defects in this model.

Farhadieh, R. D., S. Nicklin, et al. (2003). "The role of nerve growth factor and brain-derived neurotrophic factor in inferior alveolar nerve regeneration in distraction osteogenesis." J Craniofac Surg 14(6): 859-65.
 SUMMARY: Distraction osteogenesis (DO) has become the mainstay of treatment of mandibular hypoplasias. Despite the clinical acceptance of the technique in the last decade, little is known of the biological mechanism of bone and soft tissue regeneration. The biological response of peripheral nerves to distraction has not been well documented. This study examined the role of two neurotrophic molecules, nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF), in DO on nerve regeneration of the inferior alveolar nerve (IAN) in an ovine mandible model. Twelve animals were randomly divided into three groups and distracted at 5, 10, and 15 days using a mandibular osteotomy and uniaxial external distractor. The mental nerves and the IAN from the distracted site were harvested at the end of the distraction period and examined for the presence of NGF and BDNF using immunohistochemistry. Nerve growth factor expression was increased at both sites, whereas BDNF was only expressed at the mental nerve on the distracted side. Nerve growth factor and BDNF are involved in the response of the peripheral nerves to injury. Mechanical force applied to the IAN by distraction may lead to detachment of Schwann cells from their axons, leading to segmental degeneration. The resulting myelin sheath debris may serve as a trigger for higher expression of NGF and BDNF, facilitating Schwann cell proliferation and remyelination of the affected segment. Distraction of the mandible may serve as a source of subacute injury to the IAN and influence NGF and BDNF.

Fernandez, E., F. Di Rocco, et al. (2003). "Reinnervation of extraocular muscles by facial-to-oculomotor nerve anastomosis in rats: anatomic nuclear changes." Neurosurgery 53(2): 409-14; discussion 414-5.
 OBJECTIVE: Oculomotor nerve palsy greatly impairs the patient's daily life. After oculomotor nerve injury, when the central nerve stump is not available, neurotization of the distal nerve stump with a donor nerve may be performed. Here, we present an experimental anatomic study in rats related to the motor nuclear organization after facial-to-oculomotor nerve anastomosis. METHODS: In adult rats, the right oculomotor nerve was transected at the skull base. Then, the ipsilateral facial nerve was exposed at the stylomastoid foramen and connected side-to-end to one extremity of a peroneal nerve autograft. The other extremity of the nerve autograft was connected end-to-end to the distal stump of the transected oculomotor nerve. Twelve weeks later, axonal regeneration in the autograft and brainstem somatotopic representation of the reinnervated extraocular muscles were investigated by use of histological and retrograde axonal tracing techniques. RESULTS: The autograft was reinnervated by a large number of small axons, 1 to 5 microm in diameter. After tracer injection into the superior rectus and medial rectus muscles, retrogradely labeled neurons were seen not only in the ipsilateral facial nucleus (16%) but also in the contralateral nucleus (8%). Labeled neurons were also seen in the ipsilateral abducens (12%), motor trigeminus (7%), trochlear (23%), and contralateral trochlear (34%) nuclei. In normal rats, the extraocular muscles are innervated by unilateral-ipsilateral brainstem motor nuclei, except for the superior rectus and superior oblique muscles, which are innervated by bilateral, primarily contralateral, nuclei. CONCLUSION: The central rearrangement of the extraocular muscle nuclei after facial-to-oculomotor nerve anastomosis represents an original example of plasticity. Functional studies are needed to demonstrate whether this procedure might serve to restore some degree of eye motility.

Fernandez-Espejo, E., F. El Banoua, et al. (2003). "[Natural "dopaminotrophic" cell transplant: a new concept in antiparkinsonian therapy]." Rev Neurol 36(6): 540-4.
 AIM AND METHOD: Parkinson s disease is caused by the degeneration of dopaminergic neurons of substantia nigra projecting to striatum. Cellular substitution represents a potentially treatment once beneficial levodopa effects wear off. A promising therapeutic approach is grafting cells or other vectors which release neuroprotective molecules that stimulate regeneration in the damaged nigrostriatal system or, in other words, that exert a dopaminotrophic action. We have tested the suitability of intrastriatal grafts of extra adrenal chromaffin cells taken from the Zuckerkandl s organ. This paraganglion contains chromaffin cells that express and release glial cell line derived neurotrophic factor (GDNF) and transforming growth factor b1 (TGF b1), both known to protect dopamine cells in vitro and in vivo. Grafts induced a functional recovery of parkinsonian rats which developed over months. The beneficial effects of grafts of the Zuckerkandl s organ were related to long survival of grafted cells, striatal reinnervation, enhancement of dopamine levels in the host striatum, and the cell delivery into the host striatum of GDNF and TGF b1. CONCLUSION: Our result should stimulate research on the clinical applicability of transplants of the Zuckerkandl s organ in Parkinson s disease

Ferraresi, S., D. Garozzo, et al. (2003). "Common peroneal nerve injuries: results with one-stage nerve repair and tendon transfer." Neurosurg Rev 26(3): 175-9.
 The authors report their experience in the treatment of common peroneal nerve (CPN) injuries using a one-stage procedure of nerve repair and tibialis posterior tendon transfer. A series of 45 patients with traumatic injury and graft repair of the CPN is presented. From 1988 to 1991, the six patients elected for surgery had only nerve repair: five ultimately did not recover, while muscle contraction in the remaining patient was graded M1-2. Since 1991, nerve surgery in our clinic was associated with tendon transfer procedures (39 cases) which were followed by a satisfactory reinnervation rate. Nerve transection and iatrogenic injuries, torsion/dislocation of the knee, complex biosseous fractures of the leg, and gunshot wounds showed excellent to fair results in decreasing order: in nerve sections, muscle recovery scored M3 or M4+ in all the patients, and in nerve ruptures due to severe dislocation of the knee, it was M3 or M4+ in 85% of cases. The association of microsurgical nerve repair and tendon transfer has changed the course of CPN injuries.

Ferretti, A., E. Boschi, et al. (2003). "Angiogenesis and nerve regeneration in a model of human skin equivalent transplant." Life Sci 73(15): 1985-94.
 The angiogenesis and reinnervation were studied in a porcine model of human skin equivalent (SE) graft and the relationship between the two processes was investigated. Confocal laser scanning microscopy was used to monitor, during the healing process, the pattern of vascularization and reinnervation at different time points. The SE was obtained by co-culturing fibroblasts and keratinocytes on a collagen-glycosaminoglycan-chitosan biopolymer and grafted on dorsal wounds generated by full-thickness resection in 25/30 Kg Large white pigs. Frozen sections were obtained from biopsies performed in autograft and xenograft, then were immunolabeled by using the endothelial marker lectin Lactifolia and with the neuronal marker gene product PGP9.5. Cajal staining was also used to visualize the nerve fibers. The results show that the vascularization precedes the innervation process. These data are consistent with the view that the development of nervous tissue is driven by nutritional and trophic factors provided by the vascular system. The arborization of the two systems observed during the third week from the graft might play a key role in maintaining the healing process and the graft survival.

Ferretti, P., F. Zhang, et al. (2003). "Changes in spinal cord regenerative ability through phylogenesis and development: lessons to be learnt." Dev Dyn 226(2): 245-56.
 Lower vertebrates, such as fish and amphibians, and developing higher vertebrates can regenerate complex body structures, including significant portions of their central nervous system. It is still poorly understood why this potential is lost with evolution and development and becomes very limited in adult mammals. In this review, we will discuss the current knowledge on the cellular and molecular changes after spinal cord injury in adult tailed amphibians, where regeneration does take place, and in developing chick and mammalian embryos at different developmental stages. We will focus on the recruitment of progenitor cells to repair the damage and discuss possible roles of changes in early response to injury, such as cell death by apoptosis, and of myelin-associated proteins, such as Nogo, in the transition between regeneration-competent and regeneration-incompetent stages of development. A better understanding of the mechanisms underlying spontaneous regeneration of the spinal cord in vivo in amphibians and in the chick embryo will help to devise strategies for restoring function to damaged or diseased nervous tissues in mammals.

Filbin, M. T. (2003). "Myelin-associated inhibitors of axonal regeneration in the adult mammalian CNS." Nat Rev Neurosci 4(9): 703-13.
 
Fink, D. J., J. Glorioso, et al. (2003). "Therapeutic gene transfer with herpes-based vectors: studies in Parkinson's disease and motor nerve regeneration." Exp Neurol 184 Suppl 1: S19-24.
 We have examined the possibility of using herpes simplex virus (HSV)-based vectors to prevent neuronal cell death and enhance functional recovery after injury. In the 6-hydroxydopamine (6-OHDA) model of Parkinson's disease (PD) and after proximal spinal root injury, direct stereotactic injection of HSV-based vectors constructed to express the glial cell derived neurotrophic factor (GDNF) or the anti-apoptotic peptide Bcl-2 prevented neuronal death and enhanced recovery. Gene transfer may be useful in the treatment of neurologic disorders in which neuronal cell death occurs in a restricted anatomic distribution.

Fishman, H. M. and G. D. Bittner (2003). "Vesicle-mediated restoration of a plasmalemmal barrier in severed axons." News Physiol Sci 18: 115-8.
 Ca(2+)-induced endocytotic vesicles undergo protein-mediated interactions to restore a selectively permeable barrier and propagated action potentials in severed invertebrate giant axons. Similar barrier-restoration phenomena observed in cultured mammalian cells with transected neurites suggest that cellular/molecular mechanisms that repair plasmalemmal damage are phylogenetically conserved.

Flynn, L., P. D. Dalton, et al. (2003). "Fiber templating of poly(2-hydroxyethyl methacrylate) for neural tissue engineering." Biomaterials 24(23): 4265-72.
 We have developed a method to create longitudinally oriented channels within poly(2-hydroxyethyl methacrylate) (pHEMA) hydrogels for neural tissue engineering applications. Incorporated into an entubulation strategy, these scaffolds have the potential to enhance nerve regeneration after transection injuries of either the spinal cord or the peripheral nerve by increasing the available surface area and providing guidance to extending axons and invading cells. The fabrication process is straightforward and the resultant scaffolds are highly reproducible. Polycaprolactone (PCL) fibers were extruded and embedded in transparent, crosslinked pHEMA gels. Sonication of the pHEMA/PCL composite in acetone resulted in the complete dissolution of the PCL, leaving longitudinally oriented, fiber-free channels in the pHEMA gel. Regulating the size and quantity of the PCL fibers allowed us to control the diameter and number of channels. Small and large channel scaffolds were fabricated and thoroughly characterized. The small channel scaffolds had 142+/-7 channels, with approximately 75% of the channels in the 100-200 micro m size range. The large channel scaffolds had 37+/-1 channels, with approximately 77% of the channels in the 300-400 micro m range. The equilibrium water content (EWC), porosity and compressive modulus were measured for each of the structures. Small and large channel scaffolds had, respectively, EWCs of 55.0+/-1.2% and 56.2+/-2.9%, porosities of 35+/-1% and 40+/-1% and compressive moduli of 191+/-7 and 182+/-4kPa.

Fournier, A. E., B. T. Takizawa, et al. (2003). "Rho kinase inhibition enhances axonal regeneration in the injured CNS." J Neurosci 23(4): 1416-23.
 Myelin-associated inhibitors limit axonal regeneration in the injured brain and spinal cord. A common target of many neurite outgrowth inhibitors is the Rho family of small GTPases. Activation of Rho and a downstream effector of Rho, p160ROCK, inhibits neurite outgrowth. Here, we demonstrate that Rho is directly activated by the myelin-associated inhibitor Nogo-66. Using a binding assay to measure Rho activity, we detected increased levels of GTP Rho in PC12 and dorsal root ganglion (DRG) cell lysates after Nogo-66 stimulation. Rho activity levels were not affected by Amino-Nogo stimulation. Rho inactivation with C3 transferase promotes neurite outgrowth of chick DRG neurons in vitro, but with the delivery method used here, it fails to promote neurite outgrowth after corticospinal tract (CST) lesions in the adult rat. Inhibition of p160ROCK with Y-27632 also promotes neurite outgrowth on myelin-associated inhibitors in vitro. Furthermore, Y-27632 enhances sprouting of CST fibers in vivo and accelerates locomotor recovery after CST lesions in adult rats.

Francel, P. C., K. S. Smith, et al. (2003). "Regeneration of rat sciatic nerve across a LactoSorb bioresorbable conduit with interposed short-segment nerve grafts." J Neurosurg 99(3): 549-54.
 OBJECT: This study was conducted to evaluate peripheral nerve regeneration through a conduit composed of a bioresorbable material (LactoSorb). METHODS: Sprague-Dawley rats weighing approximately 250 g were randomized into five groups. A 20-mm-long sciatic nerve gap was created, then it was bridged by a reverse nerve autograft (Group I), an empty silicone tube (Group II), a silicone tube containing a short (2-mm) interposed nerve segment (Group III), an empty LactoSorb conduit (Group IV), or a LactoSorb conduit containing a 2-mm interposed nerve segment (Group V). The intact sciatic nerve served as the control in each animal. At 16 weeks postoperatively, no nerve regeneration was observed through either the empty silicone tube or the empty LactoSorb conduit. There was regeneration in all animals receiving the reverse autograft as well as in all animals receiving the silicone or LactoSorb conduit containing the 2-mm interposed nerve segment. Effective regeneration was assessed based on histological, electrophysiological, and morphometric criteria. CONCLUSIONS: The results indicate that a conduit made of resorbable material will support sciatic nerve regeneration over a critical gap defect.

Fressinaud, C., I. Jean, et al. (2003). "Selective decrease in axonal nerve growth factor and insulin-like growth factor I immunoreactivity in axonopathies of unknown etiology." Acta Neuropathol (Berl) 105(5): 477-83.
 In an attempt to approach the mechanisms underlying axonopathies of unknown etiology, we have studied by immunocytochemistry the fate of several growth factors in eight of such cases that we had previously analyzed by morphometry and which were characterized by a decrease in neurofilaments and an increase in beta tubulin immunostaining. Here we establish that, contrary to beta tubulin, growth-associated protein43 (GAP-43) immunolabeling is not up-regulated in theses cases, correlating well with the failure of regeneration. Neurotrophin-3 (NT-3) and its receptor TrkC were not modified compared to controls (five cases). On the contrary, we observed in all cases a pronounced decrease in the number of fibers labeled for nerve growth factor (NGF) and insulin-like growth factor I (IGF-I), which were both approximately half of control values. This decrease could not be ascribed to the reduction in fiber density since it was also present in cases without fiber loss (isolated large fiber atrophy). The fact that only around 50% of fibers were stained, versus all fibers in controls, probably accounted for this decrease. It contrasted also with the normality of NGF and IGF-I immunolabeling in six cases of chronic inflammatory demyelinating neuropathy that were investigated in parallel. These results differ from those reported in experimental diabetic neuropathy, during which NT-3 is also decreased. A deficient supply of specific growth factors delivered by neuronal targets may be responsible for these neuropathies and their associated axonal cytoskeleton abnormalities.

Friedman, S. M. (2003). "Optociliary venous anastomosis after radial optic neurotomy for central retinal vein occlusion." Ophthalmic Surg Lasers Imaging 34(4): 315-7.
 Radial optic neurotomy has been shown to be beneficial for the treatment of central retinal vein occlusion. Two patients developed optociliary venous anastomosis after radial optic neurotomy for central retinal vein occlusion with improvement in vision and clinical appearance. Optociliary venous anastomosis may portend a favorable prognosis.

Frost, E. E., J. A. Nielsen, et al. (2003). "PDGF and FGF2 regulate oligodendrocyte progenitor responses to demyelination." J Neurobiol 54(3): 457-72.
 Acute demyelination of adult CNS, resulting from trauma or disease, is initially followed by remyelination. However, chronic lesions with subsequent functional impairment result from eventual failure of the remyelination process, as seen in multiple sclerosis. Studies using animal models of successful remyelination delineate a progression of events facilitating remyelination. A universal feature of this repair process is extensive proliferation of oligodendrocyte progenitor cells (OPs) in response to demyelination. To investigate signals that regulate OP proliferation in response to demyelination we used murine hepatitis virus-A59 (MHV-A59) infection of adult mice to induce focal demyelination throughout the spinal cord followed by spontaneous remyelination. We cultured glial cells directly from demyelinating and remyelinating spinal cords using conditions that maintain the dramatically enhanced OP proliferative response prior to CNS remyelination. We identify PDGF and FGF2 as significant mitogens regulating this proliferative response. Furthermore, we demonstrate endogenous PDGF and FGF2 activity in these glial cultures isolated from demyelinated CNS tissue. These findings correlate well with our previous demonstration of increased in vivo expression of PDGF and FGF2 ligand and corresponding receptors in MHV-A59 lesions. Together these studies support the potential of these pathways to function in vivo as critical factors in regulating remyelination.

Fujii, N., H. Ohnishi, et al. (2003). "Regeneration of nerve fibres in the peri-implant epithelium incident to implantation in the rat maxilla as demonstrated by immunocytochemistry for protein gene product 9.5 (PGP9.5) and calcitonin gene-related peptide (CGRP)." Clin Oral Implants Res 14(2): 240-7.
 The response of nerve fibres in the peri-implant epithelium to titanium implantation was investigated with an experimental model using rat maxilla and immunohistochemical techniques. The latter employed antibodies to protein gene product 9.5 (PGP9.5), and to calcitonin gene-related peptide (CGRP). In control rats without an implantation, a dense innervation of PGP9.5- and CGRP-positive nerve fibres was recognized throughout the junctional epithelium, as has been previously reported. A titanium-implantation induced a remarkable inflammatory reaction, as well as the destruction of covering epithelial cells. By 3-5 days post-implantation, inflammatory reaction showed a tendency to disappear, and the peri-implant epithelium showed proliferation and down-growth along the implant. At this stage, no nerve fibres were found around the peri-implant epithelium. At 10 days, a few nerve fibres reached the basal cell layers of the peri-implant epithelium, and entered it 15 days after implantation when the peri-implant epithelial cells showed morphological features roughly resembling those of normal junctional epithelial cells. At the complete osseointegration stage (days 20-30), the PGP9.5- and CGRP-positive nerve fibres, thin and beaded in appearance, were found distributed in the peri-implant epithelium. After 20 days, the numerical density of the intraepithelial nerves in the peri-implant epithelium appeared the same as, or less than, that in the normal junctional epithelium. These findings indicate that the peri-implant epithelium shows the same innervation as that in normal junctional epithelium, and that the intraepithelial nerve fibres in the peri-implant epithelium might have diverse functions, which have been suggested in the literature.

Fukaya, K., M. Hasegawa, et al. (2003). "Oxidized galectin-1 stimulates the migration of Schwann cells from both proximal and distal stumps of transected nerves and promotes axonal regeneration after peripheral nerve injury." J Neuropathol Exp Neurol 62(2): 162-72.
 Oxidized galectin-1 has recently been identified as a key factor that plays important roles in initial axonal growth in injured peripheral nerves. The aim of this study was to investigate the effects of oxidized galectin-1 on regeneration of rat spinal nerves using acellular autografts (containing no viable cells) and allografts (containing no cell membranes) with special attention to the relationship between axonal regeneration and Schwann cell migration. Immunohistochemically, endogenous galectin-1 was expressed in dorsal root ganglion (DRG) neurons, spinal cord motoneurons, and axons and Schwann cells in normal sciatic nerves. Administration of oxidized recombinant human galectin-1 (rh-gal-lox, 5 ng/ml) in autograft model promoted axonal regeneration from motoneurons as well as from DRG neurons; this was confirmed by a fluorogold tracer study (p < 0.05). Anti-rh-gal-1 antibody (30 microg/ml) strongly inhibited axonal regrowth (p < 0.05). Pretreatment of allografts with rh-gal-lox stimulated the migration of Schwann cells not only from proximal stumps but also from distal stumps into the grafts, resulting in accelerated axonal regeneration (p < 0.05). Moreover, Schwann cell migration preceded the axonal growth in the presence of exogenous rh-gal-lox in the grafts. These results strongly suggest that local administration of exogenous rh-gal-lox promotes the migration of Schwann cells followed by axonal regeneration from both motor and sensory neurons, resulting in acceleration of neuronal repair. This technique may also be of value in the repair of human nerves.

Fulton, I. C., J. A. Stick, et al. (2003). "Laryngeal reinnervation in the horse." Vet Clin North Am Equine Pract 19(1): 189-208, viii.
 Left laryngeal hemiplegia is a frustrating condition for the equine athlete and equine veterinarian. Treatment for the past 30 years has centered on the prosthetic laryngoplasty ("tie-back") with or without ventriculectomy. Laryngeal reinnervation has been used successfully in people and has been shown experimentally to benefit affected horses. This article reviews equine laryngeal reinnervation using the nerve muscle pedicle graft and describes the surgical technique, its complications, and the follow-up in 146 cases treated over the past 10 years. Also discussed is ongoing research into stimulation studies to improve the success of equine laryngeal reinnervation.

Furukawa, K. and M. Nishio (2003). "[Function of gangliosides]." Tanpakushitsu Kakusan Koso 48(8 Suppl): 952-7.
 
Furukawa, Y., N. Hashimoto, et al. (2003). "Down-regulation of an ankyrin repeat-containing protein, V-1, during skeletal muscle differentiation and its re-expression in the regenerative process of muscular dystrophy." Neuromuscul Disord 13(1): 32-41.
 Using Western blot analysis and immunohistochemical methods, we examined the expression of V-1, a member of the ankyrin repeat-containing protein family, during differentiation and regeneration of skeletal muscle. The expression of V-1 was high in cultured myoblasts and decreased during their differentiation into myotubes, while high expression was maintained when muscle differentiation was inhibited by treatment with basic fibroblast growth factor. Down-regulation of V-1 also occurred during in vivo muscle differentiation from embryonic to postnatal stages, reaching an undetectable level in mature skeletal muscle. In contrast, strong V-1 immunoreactivity was detected again in myoblasts and regenerating muscle fibers with a small diameter, which were observed in Duchenne muscular dystrophy and its animal model, mdx mouse. Thus, it seems that V-1 is a good marker for early stage of muscle regeneration and changes of its expression suggest that V-1 plays a role in prenatal muscle differentiation and postnatal muscle regeneration.

Gage, F. H. (2003). "Brain, repair yourself." Sci Am 289(3): 46-53.
 
Gamez, E., K. Ikezaki, et al. (2003). "Photoconstructs of nerve guidance prosthesis using photoreactive gelatin as a scaffold." Cell Transplant 12(5): 481-90.
 We devised a novel nerve prosthesis composed of an elastomeric gelatinous tube and multifilament gelatinous fibers, both of which were prepared from styrene-derivatized gelatin, which allows in situ formation of a bioactive substance-incorporated gel. An in vitro study showed that the axonal regeneration potential of a photocured gelatin layer impregnated with laminin, fibronectin, and NGF was almost comparable with that of coated Matrigel. A nerve conduit and fibers prepared from photoreactive gelatin was subjected to visible-light irradiation with rotation in the presence of camphorquinone as a photoinitiator using a custom-designed apparatus. A sample of transparent gelatinous conduit with an inner diameter of 1.2 mm and a wall thickness of 0.6 mm and gelatin fibers ranging from 10 to 100 pm in diameter were produced. The photocured elastomeric gelatinous tube was flexible and had structural integrity that allowed mechanical handling without breaking. A novel nerve guidance prosthesis composed of tubes packed with fibers was assembled. This photofabrication technology may enable the design of a tailor-made shape and rapid morphogenesis and functional recovery of damaged nerve tissue.

Gao, M. T., D. M. Jiang, et al. (2003). "[Effect of subcutaneous implant of peripheral nerve allograft on sciatic nerve regeneration in rats]." Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi 17(4): 312-4.
 OBJECTIVE: To study the effect of subcutaneous implant of peripheral nerve allograft on sciatic nerve regeneration in rats. METHODS: Out of 30 male Wistar rats, 6 were donors and 24 were divided randomly into 2 groups. In experimental group (group A, n = 12), a 15 mm segment of sciatic nerve harvested from donors was separately inserted into subcutaneous compartment on the right thigh; two weeks later, the segment of sciatic nerve in subcutaneous compartment was removed and transplanted into a 10 mm sciatic nerve defect of left, which was made immediately. In the control group (group B, n = 12), a 10 mm sciatic nerve defect was made and immediately repaired in situ on the left thigh. The regeneration of sciatic nerve was examined histologically (after 2, 4, 8, and 14 weeks) and electrophysiologically (after 14 weeks of operation). RESULTS: After 2 weeks of operation, the inflammatory reaction was a little stronger in group A than in group B. After 4 weeks, the intensity of the inflammatory reaction was similar between two groups; some collagen fibers proliferated. After 8 weeks, the inflammatory reaction ended and the collagen fibers proliferated obviously. After 14 weeks of operation, the structure of epineurium was in integrity and there was no obvious difference in perineurium and endonurium between two groups. A large number of myelinated nerve fibers and a small number of unmyelinated nerve fibers regenerated. The structure of myelin sheath was in integrity. The number and size of regenerated axon had no significant difference between two groups(P > 0.05). The conduction velocity, the peak value and the latent period of motor nerve were no significant difference between two groups (P > 0.05). CONCLUSION: The allograft of sciatic nerve inserted into subcutaneous compartment can promote nerve regeneration.

Garcia, M., V. Forster, et al. (2003). "In vivo expression of neurotrophins and neurotrophin receptors is conserved in adult porcine retina in vitro." Invest Ophthalmol Vis Sci 44(10): 4532-41.
 PURPOSE. To characterize and compare the expression of neurotrophins (NTs) and their receptors within adult porcine retinal ganglion cells (RGCs) in vivo and in vitro. METHODS. The distribution of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3) and -4 (NT-4), and their high-affinity receptors TrkA, TrkB, TrkC and low-affinity receptor p75, was analyzed in adult porcine retinal sections by immunohistochemistry. In addition, adult porcine retinas were dissociated and cultured in four different conditions: control, semipure RGCs, supplemented with BDNF, or seeded on Muller glia feeder layers. Double immunostaining was performed with antibodies to NTs or their receptors combined with neurofilament antibody to identify RGCs in culture. RESULTS. In vivo, immunolabeling of NGF was very faint, BDNF was especially prominent in RGCs and inner nuclear layer cells, NT-3 stained widespread nuclei, and NT-4 was undetectable. TrkA immunoreactivity was visible in the nerve fiber layer, TrkB staining was within RGC bodies, TrkC was undetectable, and p75 was widely expressed across the retina, within the Muller glia. Expression of neurotrophins and their receptors was maintained in all four models of adult RGCs in vitro, showing that expression was not influenced by substrate or culture conditions. We observed prominent staining of TrkA within growth cones, and a clear expression of p75 within a subpopulation of RGCs in vitro. CONCLUSIONS. These findings demonstrate that the expression of NTs and their receptors within adult porcine RGCs is maintained in vitro, under conditions of limited interaction with neighboring neurons and deprived of afferent inputs and target tissue. TrkA may be involved in regeneration of nerve terminals.

Garland, R., F. A. Frizelle, et al. (2003). "A retrospective audit of long-term lower limb complications following leg vein harvesting for coronary artery bypass grafting." Eur J Cardiothorac Surg 23(6): 950-5.
 OBJECTIVE: To evaluate the prevalence of leg complications following leg-vein harvest for coronary artery bypass grafting. METHOD: A questionnaire was sent to patients who had coronary artery bypass surgery between January 1993 and December 1998. Questions addressed pain, numbness, infection, swelling and general healing. The relationship between the risk factors of diabetes, peripheral vascular disease, previous fractures/injuries to legs, previous deep vein clots and arthritis affecting the legs with post operative symptoms of pain, numbness, swelling and general healing was explored with multivariate analysis. RESULTS: Of 700 questionnaires sent out 497 were returned, of which 422 (60%) were suitable for analysis. Numbness or tingling related to the wound was reported by 256 (61%), of whom 94 (37%) improved within 3 months. However, 105 (41%) had persistent numbness beyond 2 years. Pain in the wound was reported by 193 (46%), of whom 149 (77%) reported that this had improved by 3 months and only 19 (10%) had pain persisting beyond 2 years. A leg wound infection was reported by 126 (32%) patients, with 82 (65%) of these receiving antibiotics. A total of 336 (87% of 387 responses) described their wound as completely healed at 3 months. Unilateral leg swelling was reported by 175 (41%) with 98 of these (56% of those with swelling) improving by 3 months and 41 (23%) with swelling persisting beyond 2 years. There was no relation of wound problems to examined risk factors diabetes (P-values for numbness 0.31, wound healing 0.15, swelling 0.21, pain 0.22) and peripheral vascular disease (P-values for numbness 0.8, wound healing 0.21, swelling 0.18, pain 0.09). There was insufficient data to comment on the influence of fractures/injuries to legs, previous deep vein clots and arthritis affecting the legs. CONCLUSIONS: Wound complications are common following leg vein harvest. Prevalence of infection was higher than has previously been reported. Few people suffer long-term pain from saphenous nerve damage although paraesthesia and swelling were common long-term complications. We did not identify either diabetes or peripheral vascular disease as a risk factor for pain, numbness, swelling or problems with general healing. There is a need for a large multicentre prospective study.

Gatzinsky, K. P., B. Holtmann, et al. (2003). "Early onset of degenerative changes at nodes of Ranvier in alpha-motor axons of Cntf null (-/-) mutant mice." Glia 42(4): 340-9.
 The nodes of Ranvier are sites of specific interaction between Schwann cells and axons. Besides their crucial role in transmission of action potentials, the nodes of Ranvier and in particular the paranodal axon-Schwann cell networks (ASNs) are thought to function as local centers in large motor axons for removal, degradation, and disposal of organelles. In order to test whether ciliary neurotrophic factor (CNTF), which is present at high levels in the Schwann cell cytoplasm, is involved in the maintenance of these structures, we have examined lumbar ventral root nerve fibers of alpha-motor neurons by electron microscopy in 3- and 9-month-old Cntf null ((-/-)) mutant mice. Nerve fibers and nodes of Ranvier in 3-month-old Cntf(-/-) mutants appeared morphologically normal, except that ASNs were more voluminous in the mutants than in wild-type control animals at this age. In 9-month-old Cntf(-/-) animals, morphological changes, such as reduction in nerve fiber and axon diameter, myelin sheath disruption, and loss of ASNs at nodes of Ranvier, were observed. These findings suggest that endogenous CNTF, in addition to its role in promoting motor neuron survival and regeneration, is needed for long-term maintenance of alpha-motor nerve fibers. The premature loss of paranodal ASNs in animals lacking CNTF, which seems to be a defect related to a disturbed interaction in the nodal region between the axon and its myelinating Schwann cells, could impede the maintenance of a normal milieu in the motor axon, preceding more general neuronal damage.

Georgiou, T., M. McKibbin, et al. (2003). "Bilateral third-nerve palsy with aberrant regeneration in Guillain-Barre syndrome." Eye 17(2): 254-6.
 
George, L. T., T. M. Myckatyn, et al. (2003). "Functional recovery and histomorphometric assessment following tibial nerve injury in the mouse." J Reconstr Microsurg 19(1): 41-8.
 Longitudinal studies have established that functional recovery following sciatic nerve injury can be evaluated in the mouse. Injury to the tibial nerve offers several advantages to sciatic nerve injury, including improved lower extremity sensation and end-organ reinnervation. Functional recovery following tibial nerve crush injury was studied in 55 C3H mice randomized into five groups harvested for histomorphometric evaluation from either normal nerves or 2, 3, 4, or 6 weeks postoperatively. Walking tracks were obtained preoperatively, and at regular intervals postoperatively, and foot print lengths measured. Significant normalization of print length occurred 14 days postoperatively, and complete recovery was noted 28 days postoperatively. Significant histomorphologic evidence of neuroregeneration was detected between 2 and 4 weeks postoperatively. Injury to the tibial nerve is a viable alternative to the sciatic nerve for studying neural regeneration in mice, and the print length factor can be used to monitor functional recovery in this model.

Gericke, C. A., U. E. Lang, et al. (2003). "Nerve growth factor response to excitotoxic lesion of the cholinergic basal forebrain is slightly impaired in aged rats." J Neural Transm 110(6): 627-39.
 Nerve growth factor (NGF) promotes survival and function of basal forebrain cholinergic neurons. We studied NGF and choline acetyltransferase (ChAT) activity after partial quisqualic acid induced lesions of the basal forebrain in 3 and 27 months-old rats, in order to investigate whether NGF-related regeneration is disturbed in old age. 2 weeks post lesion, ChAT activity decreased by 25 to 32% in adult and old rats. 3 months post lesion, the ChAT deficit receded in adult rats, but remained unchanged in old rats. 2 weeks post lesion, NGF levels were reduced by 36 to 44%, but there was no significant difference between adult and old rats. 3 months post lesion, we found increased NGF levels by 44% in the posterior cortex of adult rats. These results indicate that the compensatory NGF increase in the posterior cortex after partial cholinergic lesion of the basal forebrain is slightly impaired in old age.

Germain, F., E. Fernandez, et al. (2003). "Morphometrical analysis of dendritic arborization in axotomized retinal ganglion cells." Eur J Neurosci 18(5): 1103-9.
 It has been reported that section of the optic nerve in mammals causes death in >90% of the retinal ganglion cells (RGCs). The cells which survive the section experience an irreparable loss of many of their dendritic segments and a rapid retraction of the dendritic tree. However, some growth cones and abnormal processes have been also reported. Our aim was to make a quantitative study of the morphological changes found in rabbit RGCs after optic nerve section. The morphometrical analysis of the RGCs which survived the axotomy showed an increase in the diameter of the soma and a significant increase in the area of the dendritic field; also, the length of the dendritic segments was significantly longer in axotomized RGCs than in control cells. Terminal dendritic segments (T) and preterminal segments (PT) were both measured in control and axotomized cells; the length ratio of T : PT segments was significantly greater in the axotomized cells than in the controls. We conclude that RGCs which survived the axotomy experienced a significant growth of their terminal dendritic branches.

Gervasi, C., A. Thyagarajan, et al. (2003). "Increased expression of multiple neurofilament mRNAs during regeneration of vertebrate central nervous system axons." J Comp Neurol 461(2): 262-75.
 Characteristic changes in the expression of neuronal intermediate filaments (nIFs), an abundant cytoskeletal component of vertebrate axons, accompany successful axon regeneration. In mammalian regenerating PNS, expression of nIFs that are characteristic of mature neurons becomes suppressed throughout regeneration, whereas that of peripherin, which is abundant in developing axons, increases. Comparable changes are absent from mammalian injured CNS; but in goldfish and lamprey CNS, expression of several nIFs increases during axon regrowth. To obtain a broader view of the nIF response of successfully regenerating vertebrate CNS, in situ hybridization and video densitometry were used to track multiple nIF mRNAs during optic axon regeneration in Xenopus laevis. As in other successfully regenerating systems, peripherin expression increased rapidly after injury and expression of those nIFs characteristic of mature retinal ganglion cells decreased. Unlike the decrease in nIF mRNAs of regenerating PNS, that of Xenopus retinal ganglion cells was transient, with most nIF mRNAs increasing above normal during axon regrowth. At the peak of regeneration, increases in each nIF mRNA resulted in a doubling of the total amount of nIF mRNA, as well as a shift in the relative proportions contributed by each nIF. The relative proportions of peripherin and NF-M increased above normal, whereas proportions of xefiltin and NF-L decreased and that of XNIF remained the same. The increases in peripherin and NF-M mRNAs were accompanied by increases in protein. These results are consistent with the hypothesis that successful axon regeneration involves changes in nIF subunit composition conducive to growth and argue that a successful injury response differs between CNS and PNS.

Geuna, S., S. Raimondo, et al. (2003). "Schwann-cell proliferation in muscle-vein combined conduits for bridging rat sciatic nerve defects." J Reconstr Microsurg 19(2): 119-23; discussion 124.
 Among the various grafting procedures that have been studied as alternatives to traditional fresh nerve autografts for the repair of severed peripheral nerves, muscle-vein-combined graft conduits have recently been devised and successfully employed. In the present study, the early presence, origin, and proliferation activity of Schwann cells (SCs) along this particular type of biological graft conduit have been investigated, using antibodies directed against glial fibrillar acid protein (GFAP), a protein that is specifically expressed in glial cells, and proliferating cell nuclear antigen (PCNA), a protein that is expressed by cells during DNA synthesis. Results showed that the muscle-vein-combined graft was progressively invaded by a number of GFAP-immunopositive SCs, many of which were also found to be immunopositive for PCNA, thus demonstrating that their proliferation continues to occur inside the graft. Among the molecules that could be involved in the stimulation of Schwann-cell proliferation is neuregulin-1 (NRG-1) that mediates its effects by binding to the ErbB receptor tyrosine kinase family. In the present study, the authors report on the RT-PCR analysis for NRG-1 and ErbB3 mRNAs, showing an overall increase in the content of these transcripts inside the muscle-vein-combined graft. These results suggest that the muscle-vein-combined graft conduit constitutes an environment favorable to potentiate Schwann-cell proliferation during the early regeneration phases.

Giampaoli, S., S. Bucci, et al. (2003). "Expression of FGF2 in the limb blastema of two Salamandridae correlates with their regenerative capability." Proc R Soc Lond B Biol Sci 270(1530): 2197-205.
 Limb regenerative potential in urodeles seems to vary among different species. We observed that Triturus vulgaris meridionalis regenerate their limbs significantly faster than T. carnifex, where a long gap between the time of amputation and blastema formation occurs, and tried to identify cellular and molecular events that may underlie these differences in regenerative capability. Whereas wound healing is comparable in the two species, formation of an apical epidermal cap (AEC), which is required for blastema outgrowth, is delayed for approximately three weeks in T. carnifex. Furthermore, fewer nerve fibres are present distally early after amputation, consistent with the late onset of blastemal cell proliferation observed in T. carnifex. We investigated whether different expression of putative blastema mitogens, such as FGF1 and FGF2, in these species may underlie differences in the progression of regeneration. We found that whereas FGF1 is detected in the epidermis throughout the regenerative process, FGF2 onset of expression in the wound epidermis of both species coincides with AEC formation and initiation of blastemal cell proliferation, which is delayed in T. carnifex, and declines thereafter. In vitro studies showed that FGF2 activates MCM3, a factor essential for DNA replication licensing activity, and can be produced by blastemal cells themselves, indicating an autocrine action. These results suggest that FGF2 plays a key role in the initiation of blastema growth.

Gibran, N. S., R. Tamura, et al. (2003). "Human dermal microvascular endothelial cells produce nerve growth factor: implications for wound repair." Shock 19(2): 127-30.
 Following cutaneous injury, sensory nerves regenerate into the dermis and epidermis. Tissues that are innervated by sensory nerves synthesize neurotrophins such as nerve growth factor (NGF). The close anatomic proximity of nerves and capillaries throughout the skin suggests that mutual regulation may exist between nerve fibers and microvascular endothelial cells (MECs) during wound repair. Release of the neuropeptide substance P by sensory nerves induces endothelial cell rounding, capillary leak, and cytokine upregulation. We propose that dermal endothelial cells produce neurotrophins required for nerve fiber maintenance and regeneration. In this study, we demonstrate that substance P stimulates NGF messenger RNA expression by cultured human dermal MECs. Likewise, enzyme-linked immunosorbant assay demonstrated that conditioned medium from cultured dermal MECs contains NGF. NGF bioactivity in the supemates was verified by conditioned medium-induced clonal rat pheochromocytoma (PC-12) cell differentiation. This activity was inhibited by anti-NGF antibodies. Therefore, we have demonstrated that substance P, an inflammatory neuropeptide released by sensory nerve fibers, induces endothelial cells to produce NGF. Our data suggest that MECs may be unrecognized contributors to nerve regeneration after cutaneous injury.

Gilhuis, H. J., C. H. Beurskens, et al. (2003). "Contralateral reinnervation of midline muscles in nonidiopathic facial palsy." J Clin Neurophysiol 20(2): 151-4.
 The purpose of this study was to analyze contralateral reinnervation of the facial nerve in eight patients with complete facial palsy after surgery or trauma and seven healthy volunteers. All patients had contralateral reinnervation of facial muscles as demonstrated by electrical nerve stimulation versus none of the control subjects. Four patients had facial muscle movements at the site of the damaged nerve. In one patient this was entirely the result of contralateral reinnervation, whereas the other three patients had innervation both ipsilaterally and contralaterally. This implies that renewed facial muscle activity should be examined considering the origin of the reinnervation, either contralateral or ipsilateral. Contralateral reinnervation is a common phenomenon after total facial palsy and can occur alongside ipsilateral reinnervation. It can be mistaken for adequate reinnervation of the damaged nerve, causing postponement of dynamic reconstruction therapy.

Gillon, R. S., Q. Cui, et al. (2003). "Effects of immunosuppression on regrowth of adult rat retinal ganglion cell axons into peripheral nerve allografts." J Neurosci Res 74(4): 524-32.
 Analysis of the effectiveness of allografts and immunosuppression in the repair of nerve defects in the adult peripheral nervous system (PNS) has a long experimental and clinical history. There is little information, however, on the use of allografts in peripheral nerve (PN) transplantation into the injured central nervous system (CNS). We assessed the ability of PN allografts (from Dark-Agouti rats) to support regeneration of adult rat retinal ganglion cell (RGC) axons in immunosuppressed host Lewis rats. PN allografts were sutured onto intraorbitally transected optic nerves. Three weeks after grafting, regenerating RGC axon numbers were determined using retrograde fluorescent labelling, and total axons within PN grafts were assessed using pan-neurofilament immunohistochemistry. In the absence of immunosuppression, PN allografts contained few axons and there were very few labelled RGC. These degenerate grafts contained many T cells and macrophages. Systemic (intraperitoneal) application of the immunosuppressants cyclosporin-A or FK506 reduced cellular infiltration into allografts and resulted in extensive axonal regrowth from surviving RGCs. The average number of RGCs regenerating axons into immunosuppressed allografts was not significantly different from that seen in PN autografts in rats sham-injected with saline. Many pan-neurofilament-positive axons, a proportion of which were myelinated, were seen in immunosuppressed allografts, particularly in proximal regions of the grafts toward the optic nerve-PN interface. This study demonstrates that PN allografts can support axonal regrowth in immunosuppressed adult hosts, and points to possible clinical use in CNS repair.

Gingras, M., J. Bergeron, et al. (2003). "In vitro development of a tissue-engineered model of peripheral nerve regeneration to study neurite growth." Faseb J 17(14): 2124-6.
 A unique tissue-engineered model of peripheral nerve regeneration was developed in vitro to study neurite outgrowth. Mouse dorsal root ganglia neurons were seeded on a collagen sponge populated with human endothelial cells and/or human fibroblasts. Addition of nerve growth factor (NGF; 10 ng/ml) was not required for sensory neurons survival but was necessary to promote neurite outgrowth, as assessed by immunostaining of the 150 kDa neurofilament. A vigorous neurite elongation was detected inside the reconstructed tissue after 14 and 31 days of neurons culture, reaching up to 770 microm from day 14. Axons were often observed closely associated with the capillary-like tubes reconstructed in the model, in a similar pattern as in the human dermis. The presence of endothelial cells induced a significant increase of the neurite elongation after 14 days of culture. The addition of human keratinocytes totally avoided the twofold decrease in the amount of neurites observed between 14 and 31 days in controls. Besides the addition of NGF, axonal growth did not necessitate B27 supplement or glial cell coculture to be promoted and stabilized for long-term culture. Thus, this model might be a valuable tool to study the effect of various cells and/or attractive or repulsive molecules on neurite outgrowth in vitro.

Gingras, M., I. Paradis, et al. (2003). "Nerve regeneration in a collagen-chitosan tissue-engineered skin transplanted on nude mice." Biomaterials 24(9): 1653-61.
 A reconstructed skin made of a collagen-chitosan sponge seeded with human fibroblasts and keratinocytes and grown in vitro for 31 days was developed for the treatment of deep and extensive burns. The aim of this study was to assess whether this tissue-engineered skin could promote nerve regeneration in vivo, since recovery of sensation is a major concern for burnt patients. The human reconstructed skin was transplanted on the back of nude mice and the growth of nerve fibres within it was assessed 40, 60, 90 and 120 days after graft. Nerve growth was monitored by confocal microscopy using immunohistochemical staining of PGP 9.5 and 150 kD neurofilament, while Schwann cell migration was observed using protein S100 expression and laminin deposition. Nerve growth was first detected 60 days after transplantation and was more abundant 90 and 120 days after graft. Linear arrangements of Schwann cells were observed in the graft as early as 40 days after graft. Nerve growth was observed along these Schwann cell extensions 60 days after transplantation. We conclude that the three-dimensional architecture of the collagen-chitosan tissue-engineered skin sponge encourages nerve growth. This result provides new perspectives to increase nerve regeneration within the tissue-engineered skin by linkage of neurotrophic factors in the sponge before transplantation.

Glockner, F. and T. G. Ohm (2003). "Hippocampal apolipoprotein D level depends on Braak stage and APOE genotype." Neuroscience 122(1): 103-10.
 Apolipoprotein (APO, gene; apo, protein) D, a member of the lipocalin family, has been implicated in several, pathological conditions but neither its physiologic function(s) nor ligand(s) has been clearly identified so far. Presuming a role in nerve de- and regeneration, several groups investigated apoD alterations in Alzheimer's disease (AD). Reported data, however, were not unanimous. We determined apoD protein levels in the hippocampus in a large, carefully matched autopsy case sample. ApoD levels were compared with the severity of neuropathological changes as determined by the Braak classification and with APOE genotype, a major risk factor for developing AD. ApoD was found to be related to the severity of AD-related neurofibrillary (NF) changes and not to old age alone. No correlation was found to amyloid deposits. Brain samples with widespread NF changes showed significantly higher apoD than cases with low Braak stages. This increase, however, was restricted to the APOE epsilon3/3 group, whereas the APOE epsilon4 group did not show significant variations in hippocampal apoD.

Gojo, S. and A. Umezawa (2003). "Plasticity of mesenchymal stem cells--regenerative medicine for diseased hearts." Hum Cell 16(1): 23-30.
 The phenomenon of regeneration is of growing interest to medical researchers. Until recently this was an area in which research in flatworms and newts predominated, but there is now a proliferation of research concerning regeneration in virtually all of the organs, not only the heart. One of the object is restoration of function to a failing heart through cell transplantation, and there have been many reports seeking donor sources of somatic stem cells, i.e.: stem cells in marrow or skeletal muscle and ES cells, beginning with those in embryonic myocardial cell transplant experiments. In particular, reports of mesenchymal stem cell differentiation into nerve cell, myocardial cell, skeletal muscle cell, and vascular endothelial cell series have drawn attention to cell plasticity, and clinical applications are awaited.

Gold, B. G. and J. E. Villafranca (2003). "Neuroimmunophilin ligands: the development of novel neuroregenerative/ neuroprotective compounds." Curr Top Med Chem 3(12): 1368-75.
 FK506 (tacrolimus), initially developed as an immunosuppressant drug, represents a class of compounds with potential high impact for the treatment of human neurological disorders. While immunosuppression is mediated by the 12-kD FK506-binding-protein (FKBP-12), the neurite elongation activity of FK506 involves FKBP-52 (also known as FKBP-59 or Hsp-56), a component of mature steroid receptor complexes: FKBP-52 binds to Hsp-90, which bind to p23 and the steroid receptor protein to form the complex. The brief review focuses on how three classes of compounds (FK506 derivatives, steroid hormones, and ansamycin anti-cancer drugs, e.g., geldanamycin) increase neurite elongation/nerve regeneration (axonal elongation). A model is presented whereby neurite elongation is elicited by compounds that bind to steroid receptor chaperone proteins (e.g., FKBP-52 and Hsp-90) and thereby disrupt mature steroid receptor complexes (comprising FKBP-52, Hsp-90 and p23 in addition to the steroid receptor binding protein). Disruption of the complex leads to a "gain-of-function" whereby one or more of these steroid receptor chaperone proteins (i.e, FKBP-52, Hsp-90 or p23) activates mitogen-associated protein (MAP) kinase/extracellular signal-regulated kinase (ERK) pathway. Thus, the neurotrophic actions of these distinct classes of compounds can be understood from their ability to bind steroid receptor chaperones, thereby providing a unique receptor-mediated means to activate the ERK pathway. These studies thereby shed new light on the intrinsic mechanism regulating axonal elongation. Furthermore, this mechanism may also underlie calcineurin-independent neuroprotective actions of FK506. We suggest that components of steroid receptor complexes are novel targets for the design of neuroregenerative/neuroprotective drugs.

Goldberg, J. L. (2003). "How does an axon grow?" Genes Dev 17(8): 941-58.
 
Gordon, T., O. Sulaiman, et al. (2003). "Experimental strategies to promote functional recovery after peripheral nerve injuries." J Peripher Nerv Syst 8(4): 236-50.
 The capacity of Schwann cells (SCs) in the peripheral nervous system to support axonal regeneration, in contrast to the oligodendrocytes in the central nervous system, has led to the misconception that peripheral nerve regeneration always restores function. Here, we consider how prolonged periods of time that injured neurons remain without targets during axonal regeneration (chronic axotomy) and that SCs in the distal nerve stumps remain chronically denervated (chronic denervation) progressively reduce the number of motoneurons that regenerate their axons. We demonstrate the effectiveness of low-dose, brain-derived neurotrophic and glial-derived neurotrophic factors to counteract the effects of chronic axotomy in promoting axonal regeneration. High-dose brain-derived neurotrophic factor (BDNF) on the other hand, acting through the p75 receptor, inhibits axonal regeneration and may be a factor in stopping regenerating axons from forming neuromuscular connections in skeletal muscle. The immunophilin, FK506, is also effective in promoting axonal regeneration after chronic axotomy. Chronic denervation of SCs (>1 month) severely deters axonal regeneration, although the few motor axons that do regenerate to reinnervate muscles become myelinated and form enlarged motor units in the reinnervated muscles. We found that in vitro incubation of chronically denervated SCs with transforming growth factor-beta re-established their growth-supportive phenotype in vivo, consistent with the idea that the interaction between invading macrophages and denervated SCs during Wallerian degeneration is essential to sustain axonal regeneration by promoting the growth-supportive SC phenotype. Finally, we consider the effectiveness of a brief period of 20 Hz electrical stimulation in promoting the regeneration of axons across the surgical gap after nerve repair.

Gotz, M. and D. Steindler (2003). "To be glial or not-how glial are the precursors of neurons in development and adulthood?" Glia 43(1): 1-3.
 
Gruart, A., M. Streppel, et al. (2003). "Motoneuron adaptability to new motor tasks following two types of facial-facial anastomosis in cats." Brain 126(Pt 1): 115-33.
 The ability of the facial motor system to adapt to a new motor function was studied in alert cats after unilateral transection, 180 degrees rotation and suture of the zygomatic nerve, or transection and cross-anastomosis of the proximal stump of the buccal nerve to the distal stump of the zygomatic nerve. These procedures induced reinnervation of the orbicularis oculi (OO) muscle by different OO- or mouth-related facial motoneurons. Eyelid movements and the electromyographic activity of the OO muscle were recorded up to 1 year following the two types of anastomosis. Animals with a zygomatic nerve rotation recovered spontaneous and reflex responses, but with evident deficits in eyelid kinematics, i.e. the proper regional distribution of OO motor units was disorganized by zygomatic nerve rotation and resuture, producing a permanent defect in eyelid motor performance. Following buccal-zygomatic anastomosis, the electrical activity of the OO muscle was recovered after 6-7 weeks, but air puff-, flash- and tone-evoked reflex blinks never reached the control values on the operated side. Electromyographic OO activities and lid movements corresponding to licking and deglutition activities were observed on the operated side in buccal-zygomatic anastomosed animals up to 1 year following surgery. Mouth-related facial motoneurons did not readapt their discharges to the kinetic, timing and oscillatory properties of OO muscle fibres. A significant hyper-reflexia was observed following both types of nerve repair in response to air puffs, but not to light flashes or tones. In conclusion, adult mammal facial premotor circuits maintain their motor programmes when motoneurons are induced to reinnervate a foreign muscle, or even a new set of muscle fibres.

Gschwendtner, A., Z. Liu, et al. (2003). "Regulation, cellular localization, and function of the p75 neurotrophin receptor (p75NTR) during the regeneration of facial motoneurons." Mol Cell Neurosci 24(2): 307-22.
 The common neurotrophin receptor (p75NTR) is a member of the tumor necrosis factor receptor superfamily and binds the neurotrophins nerve growth factor, brain derived neurotrophic factor, neurotrophin-3, and neurotrophin-4. P75NTR is expressed on developing motoneurons and is reexpressed on adult motoneurons under pathological conditions such as nerve trauma or neurodegeneration. Here we examined the regulation and function of p75NTR during regeneration after peripheral transection of the facial nerve of adult mice. Axotomy led to a strong increase in p75NTR immunoreactivity on the injured and regenerating facial motoneurons and on denervated Schwann cells. Cellular colocalization also revealed p75NTR immunoreactivity on neighboring blood vessels and cells in the injured nerve, but not on activated GFAP+ astrocytes or alphaMbeta2+ microglia and macrophages. To determine the function of this receptor we examined the effects of p75NTR deficiency on neuroglial activation, on the speed of axonal regeneration, and on neuronal survival after facial axotomy in two different transgenic mouse lines carrying targeted insertions exon 4 (p75e4-/-) or exon 3 (p75e3-/-) of the p75NTR gene. In both animal models absence of p75NTR led to a twofold, early increase in the number of CD3+. T-cells and in the microglial immunoreactivity for the alpha5beta1, alpha6beta1, and alphaMbeta2 integrins at day 4 in the facial nucleus and in the crushed facial motor nerve. No changes were observed in the number of reactive GFAP+ astrocytes or on late microglial and lymphocyte responses. The rate of axonal elongation in the crushed facial nerve, as well as neuronal survival, was found to be unaffected. Overall, the current study shows that the p75NTR receptor plays an important regulatory role in early neuroglial and immune activation.

Grutzner, P. A., W. Haase, et al. (2003). "[Comparative examination between an angle stable, monokortikal osteosynthesis technique with the conventional plate osteosynthesis at the anterior arm shaft fracture]." Unfallchirurg 106(2): 121-6.
 With a retrospective Follow-up essay under the use of Matched peer variables the wellbeing course was examined by patient pairs (128 patients) assigned to 64 individually with fractures of the anterior arm shaft. A group which treats others with 2.7 or 3.5 mm DCP/LC-DCP got with the AO Point Contact basic gate (PC-Fix).The well-being course data count after the implantation on a period of 18 months.The Follow-up-rate was 100% for this time period.The patients became for each other on reason of the criteria: assigned to fracture classification, soft partial loss, accompanying injuries and age.Possible complications were: implantation conditional nerve damages, infections, implant failures, delayed healings, pseudarthroses, motion reductions and synostoses.Complications appeared (PC-Fix at 13 patients: respect, DCP: 5).The statistical testing didn't yield any statistically significant advantage for one for the two implants at a p-value of 0.5811 for.

Guntinas-Lichius, O. and C. Wittekindt (2003). "The role of growth factors for disease and therapy in diseases of the head and neck." DNA Cell Biol 22(9): 593-606.
 Growth factors are a large family of polypeptide molecules that regulate cell division in many tissues by autocrine or paracrine mechanisms. Depending on what receptors are activated, growth factors can initiate mitogenic, antiproliferative, or trophic effects, that is, growth factors act as positive or negative modulators of cell proliferation. Therefore, growth factors do not only play an important role in embryonic development and adult tissue homeostasis, but also in pathological situations like infection, wound healing, and tumorigenesis. Consequently, the application of growth factors, or vice versa the application of substances which are directed against growth factors like antigrowth factor antibodies, may have therapeutic applications. This review provides a brief account of what we know regarding growth factors in otorhinolaryngology, particularly in the field of otology, wound healing, oncology, peripheral nerve regeneration, and rhinology.

Gupta, R., Y. M. Lin, et al. (2003). "Macrophage recruitment follows the pattern of inducible nitric oxide synthase expression in a model for carpal tunnel syndrome." J Neurotrauma 20(7): 671-80.
 Chronic nerve compression (CNC) induces a permeability change in neural vasculature. As recent evidence has shown that an alteration in reactive oxidative species (ROS) is related to neural degradation and regeneration, we evaluated whether inducible nitric oxide synthase (iNOS) plays a role in a rat model for CNC. Semi-quantitative analysis of iNOS mRNA and protein were performed with in situ hybridization and immunohistochemistry, respectively, at 3, 5, and 9 months post-operatively. At 3 months, iNOS mRNA was up-regulated in the perineurium of the proximal nerve with detectable changes in compressed and distal nerve segments. This expression continued to increase in the perineurium of 5-month proximal and compressed nerve segments with distal nerve demonstrating only a slight up-regulation of iNOS mRNA. At 9 months, iNOS mRNA expression was observed in both compressed and distal nerve. iNOS protein expression followed the same pattern of iNOS mRNA. As the perineurium is the blood-nerve barrier, the data suggests that these changes maybe mediated at the level of the perineurium. As macrophages release iNOS, we also evaluated whether macrophage recruitment followed the same pattern as iNOS expression. The results of ED-1 immunostaining for macrophages indicate that macrophages were localized to the outer one-third of cross sections during early time points. At later time points, macrophages were distributed diffusely throughout the nerve sections. Contrary to Wallerian degeneration, which elicits a relatively immediate signal for macrophage recruitment, CNC provides a slow, sustained stimulus for macrophage recruitment, which may be responsible for the up-regulation of iNOS gene expression.

Gupta, R. and O. Steward (2003). "Chronic nerve compression induces concurrent apoptosis and proliferation of Schwann cells." J Comp Neurol 461(2): 174-86.
 Chronic nerve compression (CNC), as in carpal tunnel syndrome, is a common cause of peripheral nerve dysfunction in humans. Previous studies using animal models have demonstrated progressive demyelination and a slowing of nerve conduction velocity. To characterize the Schwann cell response to CNC, we evaluated total Schwann cell number, apoptosis, and proliferation in an animal model of CNC. Design-based stereologic techniques revealed a striking transient increase in Schwann cell number following CNC. Schwann cell number increased sixfold relative to the normal nerve at the site of compression at 1 month and then slowly declined toward control levels. Nevertheless, assays of apoptosis (TUNEL and an antipoly-ADP-ribose polymerase labeling assays) revealed extensive Schwann cell apoptosis at 2 weeks postcompression, which is during the time when Schwann cell number was increasing. Electron microscopic analysis confirmed that these dramatic changes in Schwann cells occurred in the absence of axon degeneration and axonal swelling and before there were any detectable alterations in nerve conduction velocity. Counts of bromodeoxyuridine-labeled Schwann cells revealed that proliferation occurred concurrently with ongoing apoptosis. To define further the possible mitogenic properties of mechanical stimuli on Schwann cells, we used an in-vitro model to deliver shear stress in the form of laminar fluid flow to pure populations of Schwann cells and confirmed that mechanical stimuli induce Schwann cell proliferation. Our findings indicate that chronic nerve compression induces Schwann cell turnover with minimal axonal injury and support the idea that mechanical stimuli have a direct mitogenic effect on Schwann cells.

Haase, S. C., J. M. Rovak, et al. (2003). "Recovery of muscle contractile function following nerve gap repair with chemically acellularized peripheral nerve grafts." J Reconstr Microsurg 19(4): 241-8.
 Acellular nerve grafts have emerged as a possible alternative for reconstruction of short (<2 cm) peripheral nerve gaps. Axonal regeneration has been demonstrated within the nerve constructs. However, very little work has been done to demonstrate both axonal regeneration and recovery of motor function following peripheral nerve gap repair with acellular nerve constructs. The authors hypothesized that chemically acellularized nerve grafts can support axonal regeneration and provide functional reinnervation of rat hindlimb muscles with equivalent efficiency to peripheral nerve autografts. Peroneal nerves were harvested from adult rats and chemically acellularized. Two- and 4-cm peroneal nerve gaps were reconstructed with either a cellular autograft or an acellular isograft. Functional recovery was evaluated with walking-track analyses and measurement of maximum tetanic isometric force (F 0 ) of the extensor digitorum longus (EDL) muscle. Walking-track analysis revealed no statistically significant difference in functional recovery in rats undergoing reconstruction of 2-cm nerve gaps with acellular isografts or cellular autografts. Maximum tetanic isometric force measurements revealed a 60 percent force deficit in EDL muscles reinnervated by 2-cm acellular nerve grafts, compared to cellular autografts. Four-centimeter acellular grafts failed to support any significant EDL muscle reinnervation. This study demonstrates that chemically acellularized peripheral nerve supports axonal regeneration and functional reinnervation across 2-cm nerve gaps, and may potentially serve as an appropriate scaffold for reintroducing cellular elements, adhesion molecules, or growth factors for repair of longer nerve gaps.

Hadlock, T. A., T. Sheahan, et al. (2003). "Biologic activity of nerve growth factor slowly released from microspheres." J Reconstr Microsurg 19(3): 179-84; discussion 185-6.
 Efficient and sustained delivery of neurotrophic factors to the regenerating nerve in biologically active form presents a challenge. Protein delivery in biodegradable microsphere vehicles has been difficult, based on destabilization and breakdown during both the loading and release phases. This study examines the extravasation and stability of Nerve Growth Factor (NGF) in polylactic-co-glycolic acid (PLGA) microspheres, via both ELISA and PC-12 bioassays. PLGA microspheres co-loaded with bovine serum albumin (BSA) and NGF were prepared by a water-in-oil-in-water (W/O/W) technique, using chloroform for the organic phase and 1 percent polyvinyl alcohol (PVA) for the emulsion step. Aliquots of lyophilized microspheres were incubated in double distilled water (dd H2O) at 37 degrees C, and the supernatants assayed over time for NGF activity. ELISA was utilized for quantitative determination of NGF concentration, and a PC-12 cell neurite outgrowth assay assessed biologic activity. Both ELISA and PC-12 assays demonstrated the extravasation of NGF from microspheres. Over time, the predicted concentration of NGF via the two assays differed, suggesting possible preservation of recognizable epitopes for ELISA, but loss of biologic activity. NGF can be stored and released from microspheres. Extravasation studies should include biologically relevant assays for activity.

Hagino, S., K. Iseki, et al. (2003). "Expression pattern of glypican-1 mRNA after brain injury in mice." Neurosci Lett 349(1): 29-32.
 Glypican-1, a heparan sulfate proteoglycan, is expressed in various tissues including developing and postnatal central nervous system. It serves as a receptor for heparin-binding molecules such as fibroblast growth factors (FGFs). We investigated whether glypican-1 was expressed after brain injury in adult mice. In situ hybridization study showed that glypican-1 mRNA was expressed in the region surrounding necrotic tissue, and that the signal intensity peaked 7 days after the cryo-injury. In addition, both FGF-2 and amyloid precursor protein (APP) were concurrently upregulated and colocalized with glypican-1 mRNA. Since FGF-2 and APP can bind to glypican-1 in vitro, the present study suggested that their autocrine/paracrine interactions with glypican-1 may be involved in neuronal regeneration and/or neurite-outgrowth inhibition after brain injury.

Hagino, S., K. Iseki, et al. (2003). "Slit and glypican-1 mRNAs are coexpressed in the reactive astrocytes of the injured adult brain." Glia 42(2): 130-8.
 The slit family serves as a repellent for growing axons toward correct targets during neural development. A recent report describes slit mRNAs expressed in various brain regions in adult rats. However, their functions in the adult nervous system remain unknown. In the present study, we investigated whether slit mRNAs were expressed in the cryo-injured brain, using in situ hybridization. All slit family members were expressed at the lesion. Slit2 mRNA was the most intensely expressed in the cells surrounding the necrotic tissue. A double-labeling study showed that slit2 mRNA was expressed in the glial fibrillary acidic protein (GFAP)-positive reactive astrocytes. In addition, glypican-1, a heparan sulfate proteoglycan that serves as a high-affinity receptor for Slit protein, was coexpressed with slit2 mRNA in the reactive astrocytes. These findings suggested that slit2 might prevent regenerating axons from entering into the lesion in concert with glypican-1.

Halfpenny, C. A. and N. J. Scolding (2003). "Immune-modifying agents do not impair the survival, migration or proliferation of oligodendrocyte progenitors (CG-4) in vitro." J Neuroimmunol 139(1-2): 9-16.
 Limited intrinsic myelin repair occurs in multiple sclerosis (MS), mediated by oligodendrocyte progenitors that divide and migrate into demyelinated lesions. Experimental remyelination suggests that this repair restores function and may protect axons from subsequent degeneration. Immunomodulatory drugs such as corticosteroids, interferon-beta and azathioprine are widely used in MS. However, their influence on disease progression is modest, for reasons that are not fully explained. The direct effects of these drugs on remyelination biology remain relatively unexplored. We have investigated the effect of these MS therapies on oligodendrocyte progenitors to identify whether drug treatment might directly compromise repair, either therapeutic or spontaneous. None of these drugs affected CG-4 survival, migration or proliferation.

Hanani, M., O. Ledder, et al. (2003). "Regeneration of myenteric plexus in the mouse colon after experimental denervation with benzalkonium chloride." J Comp Neurol 462(3): 315-27.
 Recent reports suggest a far greater plasticity in nerve tissue than previously believed. As the digestive tract is exposed to a variety of insults, this question is relevant to enteric nerves, but little is known about their ability to recover from damage. To address this problem, we ablated the myenteric plexus of the mouse colon with the detergent benzalkonium chloride (BAC) and followed the ensuing morphologic changes for up to 60 days by using light- and electron microscopy. We found that, 2 days after BAC application, the treated area was essentially devoid of intact nerve elements. From day 7, new nerve fibers were observed within the denervated region. This growth progressed until, at days 30-60, newly grown nerve fibers were present in most of this region, and the pattern of muscle innervation was similar to the normal one. At least part of these fibers originated at neurons within intact ganglia surrounding the denervated region. The cross-sectional area of neurons near the denervated region at day 14 was 52% greater than controls. Glial cells were closely associated with the regenerating nerve fibers. From day 14 onward, we observed undifferentiated cells and differentiating neurons in ganglia surrounding the denervated region, and by day 30, new neurons were present in the myenteric region, along with regenerating nerve fibers. We conclude that the myenteric plexus is endowed with a considerable ability of regeneration and plasticity. The results provide evidence for the presence of stem cells and for an adult neurogenesis in this plexus.

Hansson, T. and T. Brismar (2003). "Loss of sensory discrimination after median nerve injury and activation in the primary somatosensory cortex on functional magnetic resonance imaging." J Neurosurg 99(1): 100-5.
 OBJECT: The aim of this study was to assess the effects of median nerve injury and regeneration on neuronal activation in the somatosensory cortex by means of functional magnetic resonance (fMR) imaging and somatosensory evoked potentials (SSEPs). METHODS: Ten injured male patients (mean age 26 years) were examined 15 to 58 months after a total transection of the median nerve at the wrist that was repaired with epineural sutures. Two-point discrimination was lost in Digit II-III and sensory nerve conduction displayed decreased velocity (-29%) and amplitude (-84%) in the median nerve at the wrist. The fMR images were obtained during tactile stimulation (gentle strokes) performed separately on the volar surface of either Digit II-III or Digit IV-V (eight patients: two were excluded because of movement artifacts). The SSEPs were obtained using electrical stimulation proximal to the median nerve lesion. CONCLUSIONS: Patients with loss of sensory discrimination after median nerve damage and regeneration had larger areas of activation in fMR imaging near the contralateral central sulcus during tactile stimulation of the injured compared with the noninjured hand. The increase relative to the unaffected hand was 43% (p < 0.02) for Digit II-III stimulation and 46% (p < 0.02) for Digit IV-V stimulation. The SSEP data showed normal latency and amplitude. The enlarged area of cortical activation may be the result of reorganization, and it may indicate that larger cortical areas are involved in the discriminatory task after a derangement of the peripheral input.

Harada, F., N. Hoshino, et al. (2003). "The involvement of brain-derived neurotrophic factor (BDNF) in the regeneration of periodontal Ruffini endings following transection of the inferior alveolar nerve." Arch Histol Cytol 66(2): 183-94.
 The present study employed immunohistochemistry for protein gene product 9.5 (PGP 9.5) to examine the regeneration process of Ruffini endings, the primary mechanoreceptor in the periodontal ligament, in heterozygous mice with targeted disruption of the brain-derived neurotrophic factor (BDNF) gene and their littermates, following transection of the inferior alveolar nerve. When immunostained for PGP 9.5, periodontal Ruffini endings appeared densely distributed in the periodontal ligament of the heterozygous mice, but the density of the positively stained nerve fibers in the ligament was 20% lower than that in the control littermates. At 3 days after surgery, the PGP 9.5-positive neural elements had disappeared; they began to appear in the periodontal ligament of both animals at 7 days. However, the recovery pattern of the PGP 9.5-positive nerves differed between heterozygous and wild type mice, typical periodontal Ruffini endings morphologically identical to those in the control group appeared in the wild-type mice at 7 days, whereas such Ruffini endings were detectable in the heterozygous mice at 28 days, though much smaller in number. On day 28, when PGP 9.5-positive nerves were largely regenerated in wild type mice, their distribution was much less dense in the ligament of the heterozygous mice than in the non-treated heterozygous mice. The density of PGP 9.5-positive nerve fibers was significantly lower in the heterozygous mice than in wild type mice at any stage examined. These data showing that a reduced expression of BDNF causes delayed regeneration of the periodontal Ruffini endings suggest the involvement of BDNF in the regeneration process of these mechanoreceptors.

Harman, A. M., J. Rodger, et al. (2003). "PSA-NCAM is up-regulated during optic nerve regeneration in lizard but not in goldfish." Exp Neurol 182(1): 180-5.
 The addition of polysialic acid (PSA) to neural cell adhesion molecule (NCAM) facilitates axon growth. Here we use Western blots and immunohistochemistry to examine expression of PSA-NCAM during optic nerve regeneration. In lizard, retinal ganglion cell axons become transiently PSA-NCAM positive. By contrast, goldfish RGC axons are PSA-NCAM negative both in normal animals and throughout regeneration with the exception of a PSA-NCAM-positive fascicle arising from newly generated RGCs. Transient sialylation of NCAM in lizard may assist regeneration in the nonpermissive reptilian visual pathway and facilitate the reestablishment of a crude topographic map; down-regulation in the long term may contribute to the breakdown in topography. The lack of sialylation in goldfish presumably reflects the permissive nature of the substrate allowing axon regeneration and the successful reestablishment of a topographic map.

Hase, S., K. Kobayashi, et al. (2003). "Transcriptional pattern of a novel gene, expressed specifically after the point-of-no-return during sexualization, in planaria." Dev Genes Evol 212(12): 585-92.
 We have investigated sexualization of asexual worms in the planarian Dugesia ryukyuensis. During sexualization there is a point from which an animal cannot return to the asexual state (point-of-no-return). To isolate the genes related to the point-of-no-return, we performed differential screening and isolated one novel gene that was expressed specifically in yolk glands of the worms after the point-of-no-return and named it Dryg. It encoded 655 amino acids with a predicted molecular mass of 79 kDa. We performed a series of experiments using Dryg as a molecular marker in the yolk gland. At first, we monitored how the yolk gland was formed during sexualization. The expression in sexualizing worms at stage 3 is limited to a single type of cell that has characteristics of neoblasts, the totipotent somatic cells; however, the expression is observed in the yolk gland in sexualized worms. Furthermore, we monitored yolk glands for expression during regeneration. The original yolk glands seem to disappear after ablation, then new yolk glands appeared along the ventral nerve cords. Because this expression pattern looks like that of sexualizing worms at stage 3, we speculate that yolk gland cells may differentiate from neoblasts during regeneration as observed during sexualization.

Hashimoto, N., H. Yamanaka, et al. (2003). "Increased expression of 3 beta-hydroxysteroid dehydrogenase mRNA in dorsal root ganglion neurons of adult rats following peripheral nerve injury." Neurosci Lett 340(1): 45-8.
 3beta-hydroxysteroid dehydrogenase (3beta-HSD) is an enzyme that converts pregnenolone to progesterone. It has been believed that 3beta-HSD is simply a converting enzyme of female steroid hormone. Recently, 3beta-HSD expressing cells were identified in the spinal cord. Steroid synthesis in the nervous system may indicate that steroid plays a role in the nervous system. We report here the increased expression of 3beta-HSD mRNA in the dorsal root ganglion (DRG) after peripheral nerve injury using reverse transcription-polymerase chain reaction and in situ hybridization histochemistry techniques. We detected only a few 3beta-HSD signals in the naive DRG, and found that 3beta-HSD mRNA expression increased 3 days after injury and this increase was still observed at 14 days. Our results suggest that progesterone may have a role in the process against neuronal injury or in regeneration in the peripheral nervous system.

Hastings, N. B. and E. Gould (2003). "Neurons inhibit neurogenesis." Nat Med 9(3): 264-6.
 
Hatoko, M., A. Tanaka, et al. (2003). "Expression of alpha, beta, and gamma catenins in vascularized and nonvascularized nerve grafts during the regeneration process." J Reconstr Microsurg 19(4): 271-8.
 The authors investigated the expression of three kinds of catenins (alpha, beta, gamma) in vascularized and non-vascularized nerve grafts, using the rat sciatic nerve model. The vascularized nerve, 15 mm in length, was elevated with its nutrient vessels in the left hindlimb of rats. In the right hindlimb, a 15-mm segment of the sciatic nerve was elevated (resected) without the nutrient vessels as a non-vascularized nerve. Both nerves were sutured to the original site. At various periods up to 14 weeks after operation, the entire graft was removed. Expression of three catenins was detected by Western blot analysis and histochemical staining, using the antibody to each catenin. The level of beta catenin increased during nerve regeneration in both the vascularized and non-vascularized grafts, while the level of alpha and gamma catenins did not increase in both grafts. There was no difference in the level of the three catenins between the two methods of nerve grafting. Histochemical findings showed that at the 4th and 14th postoperative weeks, alpha and gamma catenins were stained diffusely in both grafts, while beta catenin was stained around the regenerating axons in both grafts. The degree of beta catenin was greater in the 4th postoperative week than in the 14th postoperative week, but no obvious difference in the degree of staining of the three catenins between two grafts was found. These results suggest that beta catenin may play a different role from alpha and gamma catenins in nerve regeneration, and that the expression of these catenins is not influenced by vascularization of the nerve graft.

Hauben, E., A. Gothilf, et al. (2003). "Vaccination with dendritic cells pulsed with peptides of myelin basic protein promotes functional recovery from spinal cord injury." J Neurosci 23(25): 8808-19.
 Injury-induced self-destructive processes cause significant functional loss after incomplete spinal cord injury (SCI). Cellular elements of both the innate (macrophage) and the adaptive (T-cell) immune response can, if properly activated and controlled, promote post-traumatic regrowth and protection after SCI. Dendritic cells (DCs) trigger activation of effector and regulatory T-cells, providing a link between the functions of the innate and the adaptive immune systems. They also initiate and control the body's response to pathogenic agents and regulate immune responses to both foreign and self-antigens. Here we show that post-injury injection of bone marrow-derived DCs pulsed with encephalitogenic or nonencephalitogenic peptides derived from myelin basic protein, when administered (either systemically or locally by injection into the lesion site) up to 12 d after the injury, led to significant and pronounced recovery from severe incomplete SCI. No significant protection was seen in DC recipients deprived of mature T-cells. Flow cytometry, RT-PCR, and proliferation assays indicated that the DCs prepared and used here were mature and immunogenic. Taken together, the results suggest that the DC-mediated neuroprotection was achieved via the induction of a systemic T-cell-dependent immune response. Better preservation of neural tissue and diminished formation of cysts and scar tissue accompanied the improved functional recovery in DC-treated rats. The use of antigen-specific DCs may represent an effective way to obtain, via transient induction of an autoimmune response, the maximal benefit of immune-mediated repair and maintenance as well as protection against self-destructive compounds.

Hayat, S., C. B. Wigley, et al. (2003). "Intracellular calcium handling in rat olfactory ensheathing cells and its role in axonal regeneration." Mol Cell Neurosci 22(2): 259-70.
 Intracellular calcium handling by rat olfactory ensheathing cells (OECs) is implicated in their support for regrowth of adult CNS neurites in a coculture model of axonal regeneration. Pretreatment of OECs with BAPTA-AM to sequester glial intracellular calcium ([Ca(2+)](i)) reduces significantly the numbers of cocultured neurons regrowing neurites. The mean resting [Ca(2+)](i) of OECs cultured alone or with neurons was 300 nM in an external solution containing 2.5 mM calcium ([Ca(2+)](o)). In high [K(+)](o) or zero [Ca(2+)](o), resting [Ca(2+)](i) significantly decreased. [Ca(2+)](i) significantly increased when [Ca(2+)](o) was increased to 20 mM, lonomycin, thapsigargin, and thimerosal increased [Ca(2+)](i), and caffeine, ryanodine, and cyclopiazonic acid were without effect. Of the receptor agonists tested, none induced a change in [Ca(2+)](i). The calcium influx induced by high [Ca(2+)](o) was blocked by La(3+) and SKF96365, partially inhibited by Cd(2+), and insensitive to Ni(2+) and nifedipine. Pretreatment of OECs with La(3+) reduced neurite regrowth in cocultures in a concentration-dependent manner over the range that blocked the non-voltage-gated calcium flux through a putative TRP-like channel, which, we propose, is activated in OEC-mediated axonal regeneration.

He, X. L., J. F. Bazan, et al. (2003). "Structure of the Nogo receptor ectodomain: a recognition module implicated in myelin inhibition." Neuron 38(2): 177-85.
 Failure of axon regeneration in the adult mammalian central nervous system (CNS) is at least partly due to inhibitory molecules associated with myelin. Recent studies suggest that an axon surface protein, the Nogo receptor (NgR), may play a role in this process through an unprecedented degree of crossreactivity with myelin-associated inhibitory ligands. Here, we report the 1.5 A crystal structure and functional characterization of a soluble extracellular domain of the human Nogo receptor. Nogo receptor adopts a leucine-rich repeat (LRR) module whose concave exterior surface contains a broad region of evolutionarily conserved patches of aromatic residues, possibly suggestive of degenerate ligand binding sites. A deep cleft at the C-terminal base of the LRR may play a role in NgR association with the p75 coreceptor. These results now provide a detailed framework for focused structure-function studies aimed at assessing the physiological relevance of NgR-mediated protein-protein interactions to axon regeneration inhibition.

He, F. C., Q. Y. Fan, et al. (2003). "Long-term result of guided nerve regeneration with an inert microporous polytetrafluoroethylene conduit." Chin J Traumatol 6(3): 145-51.
 OBJECTIVE: To evaluate the long-term outcome of Polytetrafluoroethylene (PTFE) conduit in nerve repair and to provide more evidence in view of its potential application to achieve a satisfactory functional recovery in clinical settings. METHODS: Thirty-six Wistar rats had their right sciatic nerve transected and were repaired with either conventional microsuture technique (Control group, n=18) or a PTFE conduit with a gap of 5 mm left between the nerve stumps (PTFE group, n=18). At 6 and 9 months after the operation, electrophysiological assessment and measurement of gastrocnemius muscle weight were conducted and morphology of the regenerated nerves were studied with image analysis. RESULTS: At 6 months postoperatively, the nerve conduction velocity recovered to 60.86% and 54.36% (P<0.05), and the gastrocnemius muscle weight recovered to 50.89% and 46.11% (P>0.05) in the Control group and the PTFE group respectively. At 9 months postoperatively, the recovery rate was 65.99% and 58.79% for NCV (P>0.05), and 52.56% and 47.89% for gastrocnemius muscle weight (P>0.05) in the Control group and the PTFE group respectively. Regenerated nerve fibers in the PTFE group had a regular round shape with no fragmentation, wrinkling or splitting of the myelin sheath. Image analysis revealed that the ratio of the myelin area to the total fiber area was larger at 9 months than at 6 months in both groups (P<<0.01). CONCLUSIONS: Microporous PTFE conduit may be an alternative for nerve repair allowing of guided nerve regeneration and functional recovery with no obvious adverse effect at long-term.

Helfand, B. T., M. G. Mendez, et al. (2003). "A role for intermediate filaments in determining and maintaining the shape of nerve cells." Mol Biol Cell 14(12): 5069-81.
 To date, the functions of most neural intermediate filament (IF) proteins have remained elusive. Peripherin is a type III intermediate filament (IF) protein that is expressed in developing and in differentiated neurons of the peripheral and enteric nervous systems. It is also the major IF protein expressed in PC12 cells, a widely used model for studies of peripheral neurons. Dramatic increases in peripherin expression have been shown to coincide with the initiation and outgrowth of axons during development and regeneration, suggesting that peripherin plays an important role in axon formation. Recently, small interfering RNAs (siRNA) have provided efficient ways to deplete specific proteins within mammalian cells. In this study, it has been found that peripherin-siRNA depletes peripherin and inhibits the initiation, extension, and maintenance of neurites in PC12 cells. Furthermore, the results of these experiments demonstrate that peripherin IF are critical determinants of the overall shape and architecture of neurons.

Higuchi, H., T. Yamashita, et al. (2003). "Functional inhibition of the p75 receptor using a small interfering RNA." Biochem Biophys Res Commun 301(3): 804-9.
 The neurotrophin receptor p75(NTR) mediates a wide variety of biological effects. Consistent with the function in controlling the survival and neurite formation, p75(NTR) is expressed during the developmental stages of the nervous system. Importantly, p75(NTR) is re-expressed in various pathological conditions and is suggested to contribute to the inhibition of neuronal regeneration and the death of the neurons. Here we develop a tool to knock down the expression of p75(NTR) by employing a small interfering RNA (siRNA). The siRNA for p75(NTR) effectively reduces the expression of endogenous p75(NTR) both in Schwann cells and dorsal root ganglion neurons in vitro. NGF-induced cell death in Schwann cells and the neurite retraction in DRG neurons induced by myelin-associated glycoprotein are attenuated by the siRNA. Inhibition of p75(NTR) in specific pathological conditions by the siRNA may provide a potential therapeutic agent.

Himanen, J. P. and D. B. Nikolov (2003). "Eph receptors and ephrins." Int J Biochem Cell Biol 35(2): 130-4.
 Eph receptors, the largest subfamily of receptor tyrosine kinases (RTKs), and their ephrin ligands are important mediators of cell-cell communication regulating cell attachment, shape, and mobility. Eph signaling is crucial for the development of many tissues and organs including the nervous and cardiovascular systems. Both Ephs and ephrins are membrane-bound and their interactions at sites of cell-cell contact initiate unique bi-directional signaling cascades where information is transduced in both the receptor- and the ligand-expressing cells. Recent studies summarized in this review reveal how the signaling process is triggered upon ligand-receptor binding via the formation of a 2:2 circular heterotetramer. This fixes the orientation of the participating molecules and facilitates phosphorylation of their cytoplasmic domains which then interact with downstream signaling factors. The elucidation of the structural details of Eph-ephrin recognition and binding should yield insight into the future development of novel therapeutic agents targeting cardiovascular function, nerve regeneration, and cancer.

Hiraiwa, K. (2003). "[Novel findings from an animal tourniquet shock model]." Nippon Hoigaku Zasshi 57(2): 125-34.
 This article is a review of our experimental results regarding the physiological statuses and roles of chemical mediators in tourniquet shock, and a novel phenomenon, modulation reflex, that is commonly observed in this shock model is discussed. In a rabbit with a tourniquet applied to a hind limb for 24 hrs, blood pressure (BP) gradually falls after release of the tourniquet, but the decline in BP stops when a tourniquet is again applied to the hind limb, indicating that shock mediators are attributed to the hind limb. The levels of dipeptides (anserine and carnosine) and lysosomes in blood samples as well as the levels of leukotrienes (LTD4 and LTE4) in blood and muscle samples from rabbits in tourniquet shock were elevated. However, injection of a large amount of a dipeptide into an ear vein of a rabbit did not reduce BP, suggesting that both peptides may not be directly related with reduction in BP of rabbits in tourniquet shock. Injection of a platelet-activating factor (PAF) antagonist into an ear vein resulted in slight elevation of BP and the elevated level was maintained for about 1 to 4 hrs during the period of decline in BP in tourniquet shock. As for interleukin-6 (IL-6), IL-6-deficient mice at young ages have a significantly greater blood volume than do wild-type mice without concomitant changes in body composition. Therefore, the role for IL-6 in the regulation of peripheral circulation may be to elevate, not reduce BP. In mice in tourniquet shock, superoxide (O2-) production is observed in skeletal muscle cells and these cells correspond to mitochondria-rich cells. However, RT-PCR of muscle samples showed no significant nitric oxide synthase (NOS) mRNA expression after tourniquet release. Pretreatment with NOS inhibitors before tourniquet release reduced O2- production in the skeletal muscle. These results indicate that O2- produced in muscle subjected to ischemia/repefusion may be involved in shock. As for changes in mRNA expression patterns of pro-inflammatory cytokines and nerve growth factors in blood samples from rats in tourniquet shock, up-regulation of M-CSF mRNA began at 2 h after tourniquet application and was short-lived. The level of ATF-3 mRNA had increased at 1 h and NGF mRNA gradually increased and reached a significantly high level at 4 h after tourniquet application. These results indicate that the transient mRNA expressions probably trigger secondary events that may be beneficial to wound repair and regeneration. In the early stage of tourniquet shock, the levels of IL-6 mRNA in the liver and kidneys of rats increased progressively and significantly, and the levels of iNOS mRNA in the kidneys increased. These findings suggest that that humoral and/or cellular mediators produced locally in the hind limb are responsible for remote organ injuries. Thus, these mediators, interacting each other, may contribute to the progress of shock. We have also found a novel phenomenon in tourniquet shock using rabbits. When a tourniquet is applied to the upper hind limb of a rabbit for 24 hrs, and pressure is applied to the femoral medial area immediately after tourniquet release, a reflex of decrease in blood pressure and decrease in heart rate, which last for a short period, is usually observed. This reflex is mediated through the ipsilateral femoral nerves, central nervous system and vagal nerves. Since the modulation reflex may be due to peripheral nerve injury, we investigated morphological and molecular changes in sciatic nerves and dorsal root ganglion (DRG) neurons in rats after tourniquet application. At 4 hr after tourniquet application, light microscopic examination showed only degeneration of the tourniquet segment in the sciatic nerve but no morphological changes in the DRG, while electron microscopic examination revealed mitochondrial swelling in some DRG neurons on the tourniquet-applied side and calcium deposition in these swollen mitochondria. These findings suggest that peripheral nerve injury induced a large amount of calcium influx into neuronal cell somas and that excess amounts of calcium-influx into neurons resulted in mitochondial swelling. Results of mRNA level analyses showed NGF mRNA expression followed by NGF protein expression in Schwann cells of the ipsilateral DRGs at 4 h after tourniquet application but not in the contralateral or control DRGs. Similarly, significantly high nNOS and iNOS mRNA levels were observed in the ipsilateral DRGs at 4 h, and expressions of nNOS and iNOS proteins were detected in the ganglion of the ipsilateral DRG. In addition, the TNF-alpha mRNA levels were significantly increased in the ipsilateral DRGs at 1 h after tourniquet application, indicating that TNF-alpha was activated in the early stage of nerve injury and then induced iNOS mRNA expression. Large amounts of nitric oxide (NO) produced by iNOS might result in damage to the host cells, and an overdose of NO might induce apoptosis and eliminate damaged cells during the early stage of nerve injury.

Hirakawa, H., S. Okajima, et al. (2003). "Loss and recovery of the blood-nerve barrier in the rat sciatic nerve after crush injury are associated with expression of intercellular junctional proteins." Exp Cell Res 284(2): 196-210.
 The blood-nerve barrier in peripheral nerves is important for maintaining the environment for axons. Breakdown of the barrier by nerve injury causes various pathologies. We hypothesized that the breakdown and recovery of the blood-nerve barrier after injury are associated with the changes in the expression of intercellular junctional proteins. To test this hypothesis, we induced crush injuries in the rat sciatic nerve by ligation and analyzed spatiotemporal changes of claudin-1, claudin-5, occludin, VE-cadherin, and connexin43 by immunoconfocal microscopy and morphometry and compared them with changes in the permeability of the blood-nerve barrier by intravenous and local administration of Evans blue-albumin (EBA). On day 1 after removal of the ligature EBA leaked into the connective tissue in the endoneurium and then the leakage gradually decreased and disappeared on day 7. On day 1 claudin-1, claudin-5, occludin, VE-cadherin, and connexin43 had totally disappeared from the perineurium and endoneurium. Thereafter, claudin-1, claudin-5, occludin, and VE-cadherin recovered from day 2, whereas connexin43 was redetected on day 5. These results indicate that the breakdown and following recovery of the blood-nerve barrier are closely associated with changes in the expression of claudins, occludin, VE-cadherin, and connexin43 and that the recovery time course is similar but nonidentical.

Hiroi, S., Y. Tsukamoto, et al. (2003). "Involvement of gicerin, a cell adhesion molecule, in development and regeneration of chick sciatic nerve." FEBS Lett 554(3): 311-4.
 We have examined the role of gicerin, an immunoglobulin superfamily cell adhesion molecule, in chick sciatic nerves during development and regeneration. Gicerin was expressed in the spinal cord, dorsal root ganglion (DRG) and sciatic nerves in embryos, but declined after hatching. Neurite extensions from explant cultures of the DRG were promoted on gicerin's ligands, which were inhibited by an anti-gicerin antibody. Furthermore, gicerin expression was upregulated in the regenerating sciatic nerves, DRG and dorsal horn of the spinal cord after injury to the sciatic nerve. These results indicate that gicerin might participate in the development and regeneration of sciatic nerves.

Hirata, K., J. He, et al. (2003). "HSP27 is markedly induced in Schwann cell columns and associated regenerating axons." Glia 42(1): 1-11.
 It is well known that regenerating axons enter Schwann cell (SC) columns, within which they grow to reinnervate the appropriate targets. The current study detected a marked induction of a 27-kDa heat shock protein (HSP27) in the SC columns of crush-injured rat sciatic nerves. Immunohistochemical studies showed the first appearance of strong HSP27-immunoreactive linear structures in the proximal stump near an injury site 7 h after an operation. The HSP27-immunoreactive linear structures crossed the injury site to the distal stump 2 days after the operation. They then extended in a more proximal and more distal direction and were found to have propagated through the entire length of the nerve 1 week after the operation. This pattern of expression was maintained until 3 weeks after the operation. Double-immunofluorescent labeling and confocal laser microscopy confirmed that the linear structures consisted of SC columns and associated multiple axons. The HSP27-immunoreactive SC columns expressed glial fibrillary acidic protein, but not S-100 protein. Electron microscopy and immunoelectron microscopy demonstrated that reactive Schwann cells (SCs) and the associated axons with an outgrowing profile exhibited a strong immunoreactivity to HSP27, with the former containing a greater number of bundles of intermediate filaments. It is suggested that HSP27 may play an essential role in axonal outgrowth, especially by contributing to cytoskeletal dynamics in SCs.

Hofsaess, U. and J. P. Kapfhammer (2003). "Identification of numerous genes differentially expressed in rat brain during postnatal development by suppression subtractive hybridization and expression analysis of the novel rat gene rMMS2." Brain Res Mol Brain Res 113(1-2): 13-27.
 During postnatal development the potential for axonal growth and regeneration in the central nervous system (CNS) becomes very restricted. This decline of axon growth and regeneration might be due to developmental alterations in the expression level of genes which are strongly expressed in differentiating neurons during formation of axons, but which are downregulated later in development. In order to identify genes which are downregulated in rat brain with the completion of neuronal differentiation, we performed suppression subtractive hybridization (SSH) with rat cerebellum at two developmental stages. Several differentially expressed genes were identified. We present the detailed expression analysis of one of these, rMMS2, which is the rat homologue of mouse ubiquitin-conjugating enzyme-like protein MMS2 and belongs to a family of ubiquitin-conjugating enzyme variants (UEVs) that are highly similar to ubiquitin-conjugating enzymes E2 (Ubcs) but lack the essential amino acid residue in the active site. UEVs play a role in DNA repair and are possibly involved in ubiquitination, which may be important for the assembly and function of neuronal circuits. In the present study, we examined the temporal and spatial expression of rMMS2 transcript and show a strong developmental downregulation in rat brain by Northern blot analysis and in situ hybridization. The mRNA of rMMS2 is widely distributed in rat brain at late embryonic development but is differentially regulated during postnatal development; its expression is strongly reduced during maturation of the CNS. Our results show that SSH is a suitable method for identifying genes which are regulated during postnatal development and suggest that the newly identified rat UEV rMMS2 may play a role in neuronal development and differentiation.

Holmes, M., D. Maysinger, et al. (2003). "Neotrofin, a novel purine that induces NGF-dependent nociceptive nerve sprouting but not hyperalgesia in adult rat skin." Mol Cell Neurosci 24(3): 568-80.
 We report peripheral actions in rats of Neotrofin, a purine derivative of therapeutic interest. Systemic injections mimicked NGF in eliciting sprouting of nociceptive nerves without affecting their regeneration. The sprouting was prevented by anti-NGF treatment, implicating endogenous NGF. We detected no Neotrofin-induced increases in cutaneous NGF levels or in retrograde NGF transport. In contrast, both NGF and phosphorylation of trkA increased significantly in DRGs, with a marginal appearance of phosphorylated trkA in axons. We conclude that the DRG effects of Neotrofin are responsible for its induction of sprouting. Neotrofin also induced a striking phosphorylation of axonal erk 1 and 2, which was, however, unaffected by anti-NGF treatment. We suggest that this NGF-independent MAP kinase activation is involved in nonsprouting functions of Neotrofin such as neuroprotection. Unlike injected NGF, Neotrofin did not induce hyperalgesia, supporting its candidacy as a treatment for peripheral neuropathies like those induced by diabetes and anticancer chemotherapy.

Houle, J. D. and A. Tessler (2003). "Repair of chronic spinal cord injury." Exp Neurol 182(2): 247-60.
 Advances in medical and rehabilitative care now allow the 10-12,000 individuals who suffer spinal cord injuries each year in the United States to lead productive lives of nearly normal life expectancy, so that the numbers of those with chronic injuries will approximate 300,000 at the end of the next decade. This signals an urgent need for new treatments that will improve repair and recovery after longstanding injuries. In the present report we consider the characteristics of the chronically injured spinal cord that make it an even more challenging setting in which to elicit regeneration than the acutely injured spinal cord and review the treatments that have been designed to enhance axon growth. When applied in the first 2 weeks after experimental spinal cord injury, transplants, usually in combination with supplementary neurotrophic factors, and possibly modifications of the inhibitory central nervous system environment, have produced limited long-distance axon regeneration and behavioral recovery. When applied to injuries older than 4 weeks, the same treatments have almost invariably failed to overcome the obstacles posed by the neurons' diminished capacity for regeneration and by the increasing hostility to growth of the terrain at and beyond the injury site. Novel treatments that have stimulated regeneration after acute injuries have not yet been applied to chronic injuries. A therapeutic strategy that combines rehabilitation training and pharmacological modulation of neurotransmitters appears to be a particularly promising approach to increasing recovery after longstanding injury. Identifying patients with no hope of useful recovery in the early days after injury will allow these treatments to be administered as early as possible.

Hsieh, P. S., D. J. Bochinski, et al. (2003). "The effect of vascular endothelial growth factor and brain-derived neurotrophic factor on cavernosal nerve regeneration in a nerve-crush rat model." BJU Int 92(4): 470-5.
 OBJECTIVE: To test the hypothesis that an intracavernosal injection with brain-derived neurotrophin factor (BDNF) and vascular endothelial growth factor (VEGF) can facilitate nerve regeneration and recovery of erectile function after cavernosal nerve injury. MATERIALS AND METHODS: The study included 25 Sprague-Dawley rats; four had a sham operation, seven bilateral nerve crushing with no further intervention, and 14 bilateral nerve crushing with either an immediate (seven) or delayed for 1 month (seven) intracavernosal injection with BDNF+VEGF. Erectile function was assessed by cavernosal nerve electrostimulation at 3 months, and neural regeneration by NADPH-diaphorase staining and tyrosine hydroxylase (TH) staining of penile tissue and major pelvic ganglia (MPG). RESULTS: After nerve crushing, the functional evaluation at 3 months showed a lower mean (SD) intracavernosal pressure (ICP) with cavernosal nerve stimulation, at 33.9 (15.3) cmH2O, than in the sham group, at 107.8 (18.1) cmH2O. With an immediate injection with BDNF+VEGF the ICP was significantly higher than in the controls, at 67.8 (38.5) cmH2O. Even delayed injection with BDNF+VEGF improved the ICP, to 78.0 (21.8) cmH2O. Histological analysis of specimens stained for NADPH and TH showed a significant change in the morphology of terminal branches of the cavernosal and dorsal nerves, and the staining quality of the neurones in the MPG. The number of positively stained nerve fibres tended to revert to normal after treatment with BDNF+VEGF. CONCLUSION: An intracavernosal injection with BDNF+VEGF appears to both prevent degeneration and facilitate regeneration of neurones containing neuronal nitric oxide synthase in the MPG, dorsal nerve and intracavernosal tissue. Therefore it might have therapeutic potential for enhancing the recovery of erectile function after radical pelvic surgery.

Hu, Q. L., Y. J. Piao, et al. (2003). "[Expression and distribution of NGF and p75 during rabbit tibial nerve repair induced by human hair keratin conduits]." Di Yi Jun Yi Da Xue Xue Bao 23(9): 929-32.
 OBJECTIVE: To study the expression and distribution of nerve growth factor (NGF) and low-affinity neurophin receptor p75 during human hair keratin conduit-induced repair of rabbit tibial nerves. METHODS: Rabbit tibial nerves were transected and connected by either routine suture or by conduits made of human hair keratin (HHK), and after different time periods, paraffin-embedded sections of the nerve tissue at the damaged sites and the adjacent tissues, with normal rabbit tibial nerve sections as control, were prepared for immunohistochemistry. RESULTS: No positive NGF staining was observed in normal tibial nerve tissues, but 76 days after the surgery, strong NGF positivity was detected in the newly generated nerve tissue around the HHK implants until 100 days after the surgery, which was absent in the tissues around the suture. As for p75, there was no positive staining observed in normal tibial nerve tissue. Light positive p75 staining was found in the mature nerve tissues around the HHK implants, where the newly generated tissues were strongly p75-positive during the period between 76-and loo-days after surgery. CONCLUSIONS: HHK and its degenerative product, but not routine suture, can induce the production of NGF and p75 to create a favorable micro-environment for nerve regeneration. More NGF and p75 are produced in newly generated neurons than in mature ones.

Hu, J., S. Zou, et al. (2003). "Response of Schwann cells in the inferior alveolar nerve to distraction osteogenesis: an ultrastructural and immunohistochemical study." Int J Oral Maxillofac Surg 32(3): 318-24.
 The biological mechanisms of nerve adaptation to distraction osteogenesis have not yet been elucidated. This study observed response of Schwann cells in the inferior alveolar nerve (IAN) following mandibular lengthening by electron microscopy and immunohistochemistry of S-100 protein, a specific marker of Schwann cells. Unilateral mandibular distraction (10mm elongation) was performed in nine young adult goats. Three animals were sacrificed at 7, 14 and 28 days after completion of distraction, respectively. The distracted IAN specimens and control nerves (from the contralateral sides) were harvested and processed for histological, ultrastructural and immunohistochemical examinations. Wallerian degeneration was observed in the distracted IAN, and Signs of axonal regeneration, as well as many activated Schwann cells were seen in the lengthened nerves. The expression of S-100 protein increased significantly at early stage of distraction osteogenesis, but almost returned to the normal level at 28 days after distraction. This study suggests that Wallerian degeneration caused by mechanical stretching may stimulate Schwann cells to enter a proliferated and activated state. Schwann cells and S-100 protein appear to play crucial roles in axonal regeneration that contributes to nerve adaptation to gradual distraction. Therefore, the IAN injury caused by mandibular gradual distraction was not serious; it seems to recover totally through a complicated repair mechanism.

Hua, J., V. P. Kumar, et al. (2003). "Microscopic changes at the neuromuscular junction in free muscle transfer." Clin Orthop(411): 325-33.
 Free muscle transfers do not generate the same force after transfer as that at the original sites. Light and electron microscopy were used to study serially during 30 weeks the changes at the neuromuscular junction after free muscle transfer of the gracilis muscle in the adult Wistar rat. Under light microscopy, after staining with acetylthiocholine the neuromuscular junction showed changes of degeneration with withdrawal of the innervating axon terminal followed by regeneration and reconstitution of the neuromuscular junction. The newly formed neuromuscular junction still lacked the structural detail seen in the control neuromuscular junction, even after 30 weeks. With the electron microscope, mitochondrial swelling and clumping of the synaptic vesicles were followed by withdrawal of the axon terminal from the muscle membrane on denervation. The infolding of the muscle membrane at the neuromuscular junction became less prominent. With reinnervation the ultrastructure of the junction was only partially reestablished with poorly reconstituted primary and secondary folds of the muscle membrane 30 weeks after the transfer. Failure of complete reformation of the ultrastructure of the neuromuscular junction may provide another explanation for failure of full recovery of skeletal muscle function after free muscle transfer.

Huang, M. C., K. C. Chen, et al. (2003). "Cervical root repair in adult rats after transection: recovery of forelimb motor function." Exp Neurol 180(2): 101-9.
 Functional recovery was achieved in rats after repairing the transected left sixth and seventh cervical roots. Intercostal nerves were used for reanastomosis between the transected roots and the spinal cord, and acidic fibroblast growth factor with fibrin glue was applied. Experimental rats showed relevant functional recovery of gait and grooming reflexes. Electromyography demonstrated less denervation and more regeneration. Horseradish peroxidase retrograde axonal tracing disclosed a statistically significant increase of motor neuron survival, suggesting that motor neuron survival was significantly correlated with functional recovery. It is our belief that this novel treatment strategy may help patients with similar injuries in the future.

Huang, X., D. Y. Wu, et al. (2003). "Support of retinal ganglion cell survival and axon regeneration by lithium through a Bcl-2-dependent mechanism." Invest Ophthalmol Vis Sci 44(1): 347-54.
 PURPOSE: To explore whether lithium, a long-standing mood-stabilizing drug, can be used to induce expression of Bcl-2 and support the survival and regeneration of axons of retinal ganglion cells (RGCs). METHODS: Levels of expression of Bcl-2 in the retina were assessed with quantitative reverse transcription-polymerase chain reaction. To determine whether lithium directly supports the survival of and axon-regenerative functions of RGCs, various amounts of lithium were added to cultures of isolated RGCs. Anti-Thy1.2 antibodies-conjugated to magnetic beads were used to isolate the RGCs. In addition, retina-brain slice cocultures were prepared from tissues of Bcl-2-deficient or Bcl-2-transgenic mice and treated with various amounts of lithium. The effects of the expression of Bcl-2 on lithium-mediated functions were then analyzed. RESULTS: Normal mouse retina expressed very low levels of Bcl-2 after birth. Addition of lithium in the culture increased mRNA levels of Bcl-2 in retinas of postnatal mice in a dose-dependent manner. Moreover, lithium promoted not only the survival of RGCs but also the regeneration of their axons. Depleting or forcing the expression of Bcl-2 in RGCs eliminated the effects of lithium. CONCLUSIONS: Lithium supports both the survival and regeneration of RGC axons through a Bcl-2-dependent mechanism. This suggests that lithium may be used to treat glaucoma, optic nerve neuritis, the degeneration of RGCs and their nerve fibers, and other brain and spinal cord disorders involving nerve damage and neuronal cell loss. To achieve full regeneration of the severed optic nerve, it may be essential to combine lithium therapy with other drugs that mediate induction of a permissive environment in the mature central nervous system.

Iannotti, C., H. Li, et al. (2003). "Glial cell line-derived neurotrophic factor-enriched bridging transplants promote propriospinal axonal regeneration and enhance myelination after spinal cord injury." Exp Neurol 183(2): 379-93.
 Glial cell line-derived neurotrophic factor (GDNF), a distant member of the transforming growth factor-beta (TGF-beta) family, is widely expressed in the developing and adult central nervous system (CNS). At present, limited information is available regarding the effects of GDNF in the repair of spinal cord injury (SCI). In the present study, mini-guidance channels containing either: (1) Matrigel (MG, a basement membrane component), (2) Schwann cells (SCs, 120 x 10(6)/ml) in MG (SC-MG), (3) recombinant human GDNF (rhGDNF, 3 microg/microl) in MG (GDNF-MG), and (4) a combination of all three components (GDNF-SC-MG) were grafted into a T9 hemisection-gap lesion in adult rats to examine the effects of GDNF on axonal regeneration and myelination following SCI. Thirty days post-transplantation, limited axonal growth was observed within guidance channels containing MG-alone (MG). When SCs were added to the channels (SC-MG group), consistent axonal ingrowth containing both myelinated and unmyelinated axons was observed, confirming our previous findings. The addition of GDNF-alone without SCs (GDNF-MG) resulted in substantial ingrowth of unmyelinated axons, suggesting that GDNF has a direct neurite-growth promoting effect on these axons. Implantation of channels containing both GDNF and SCs (GDNF-SC-MG) produced a significant and synergistic increase in axonal regeneration and myelination. In addition, GDNF reduced the extent of reactive gliosis, infiltration of activated macrophages/microglia, and cystic cavitation at the graft-host interfaces. Retrograde tracing revealed that grafts of SC-seeded channels containing GDNF promoted a significant increase in the number of propriospinal neurons which had regenerated their axons into the grafts, as compared to SC-MG-seeded channels. These results indicate that GDNF may play a novel therapeutic role in promoting propriospinal axonal regeneration, enhancing myelin formation, and improving graft-host interfaces after SCI.

Iijima, K., F. Harada, et al. (2003). "Temporal expression of immunoreactivity for heat shock protein 25 (Hsp25) in the rat periodontal ligament following transection of the inferior alveolar nerve." Brain Res 979(1-2): 146-52.
 The present study examined the immunohistochemical localization of heat shock protein 25 (Hsp25) during the regeneration of nerve fibers and Schwann cells in the periodontal ligament of the rat lower incisor following transection of the inferior alveolar nerve. In the untreated control group, the periodontal ligament of rat incisor did not contain any Hsp25-immunoreaction. On postoperative day 3 (PO 3d), a small number of Schwann cells with slender cytoplasmic processes exhibited Hsp25-immunoreactivity. From PO 5d to PO 21d, Hsp25-positive nerve fibers and Schwann cells drastically increased in number in the alveolar half of the ligament. Although the axons of some regenerating Ruffini-like endings also showed Hsp25-immunoreactions, the migrated Schwann cells were devoid of Hsp25-immunoreaction. Thereafter, Hsp25-positive structures decreased in number gradually to disappear from the periodontal ligament by PO 56d. This temporal expression of Hsp25 in the periodontal ligament well-reflected the regeneration process of the nerve fibers. Hsp25 in the regenerating nerve fibers and denervated Schwann cells most likely serves in modulating actin dynamics and as a cellular inhibitor of apoptosis, respectively.

Ikeguchi, R., R. Kakinoki, et al. (2003). "Rat nerve regeneration through a silicone chamber implanted with negative carbon ions." Brain Res Dev Brain Res 140(1): 127-31.
 We investigated whether a tube with its inner surface implanted with negatively-charged carbon ions (C(-) ions) would enable axons to extend over a distance greater than 10 mm. The tube was found to support nerves regenerating across a 15-mm-long inter-stump gap. Silicone treated with C(-) ions showed increased hydrophilic properties and cellular affinity, and axon regeneration was promoted with this increased biocompatibility.

Imai, T., Y. Atsumi, et al. (2003). "Regeneration of periodontal Ruffini endings of rat lower incisors following nerve cross-anastomosis with mental nerve." Brain Res 992(1): 20-9.
 The present study utilized protein gene product 9.5 (PGP 9.5) and S-100 protein immunohistochemistry to examine if Ruffini endings, the primary mechanoreceptors in periodontal ligaments, can regenerate following nerve cross-anastomosis with an inappropriate nerve. Normally, axon terminals of periodontal Ruffini endings are extensively ramified, and terminal Schwann cells, identified by their S-100 immunoreactivity, are associated with axon terminals. Schwann cells are restricted to the alveolus-related part (ARP), but not tooth-related part (TRP) or the shear zone at the border between the ARP and the TRP of the lingual periodontal ligament of the lower incisor. When the central portion of the mental nerve (MN) was connected with the peripheral portion of the inferior alveolar nerve (IAN), regenerating MN fibers invaded the IAN around postoperative day 5 (PO 5). During the postoperative period, numerous S-100-immunoreactive (IR) cells, presumably terminal Schwann cells, began to migrate to the shear zone and the TRP. PGP 9.5-IR elements reappeared at PO 7 and gradually increased in number. Around PO 28, the terminal portion of the regenerating Ruffini endings appeared dendritic, but less expanded, and the rearrangement of terminal Schwann cells was noted. Regenerated periodontal Ruffini endings were slightly smaller in number. The number of trigeminal ganglion neurons sending peripheral processes beyond the site of injury was smaller compared to those of normal MN, but their cross-sectional areas were almost comparable. Expressions of calbindin D28k and calretinin, normally localized in axonal elements in Ruffini endings, were first detected around PO 56. The present results show that parts of periodontal Ruffini endings can regenerate following nerve cross-anastomosis with mental nerve.

Imitola, J., E. Y. Snyder, et al. (2003). "Genetic programs and responses of neural stem/progenitor cells during demyelination: potential insights into repair mechanisms in multiple sclerosis." Physiol Genomics 14(3): 171-97.
 In recent years, it has become evident that the adult mammalian CNS contains a population of neural stem cells (NSCs) described as immature, undifferentiated, multipotent cells, that may be called upon for repair in neurodegenerative and demyelinating diseases. NSCs may give rise to oligodendrocyte progenitor cells (OPCs) and other myelinating cells. This article reviews recent progress in elucidating the genetic programs and dynamics of NSC and OPC proliferation, differentiation, and apoptosis, including the response to demyelination. Emerging knowledge of the molecules that may be involved in such responses may help in the design of future stem cell-based treatment of demyelinating diseases such as multiple sclerosis.

Inman, D. M. and O. Steward (2003). "Ascending sensory, but not other long-tract axons, regenerate into the connective tissue matrix that forms at the site of a spinal cord injury in mice." J Comp Neurol 462(4): 431-49.
 Mice exhibit a unique wound healing response following spinal cord injury in which the lesion site fills in with a connective tissue matrix. Previous studies have revealed that axons grow into this matrix, but the source of the axons remained unknown. The present study assesses whether any of these axons were the result of long tract regeneration. C57Bl/6 mice received crush injuries and were allowed to survive for 6 weeks to 7 months. Biotinylated dextran amine (BDA) was injected into the somato-motor cortex to trace descending corticospinal tract (CST) axons, into the midbrain to label descending brainstem pathways including the rubrospinal and reticulospinal tracts, or into the L5 dorsal root ganglion to trace ascending projections of first-order sensory neurons. Spinal cords from other mice were prepared for immunocytochemistry using antibodies against neurofilament protein (NF), 5-HT to reveal descending serotonergic axons, calcitonin gene-related protein (CGRP) to reveal ascending sensory axons, and chondroitin sulfate proteoglycan (CSPG) to assess the distribution of molecules that are inhibitory to axon growth. NF immunostaining revealed axons in the connective tissue matrix at the lesion site, confirming previous studies that used protargol staining. CST axons did not enter the connective tissue matrix, but did sprout extensively in segments adjacent to the injury site. Rubrospinal and reticulospinal tract axons also did not grow into the lesion site. 5-HT-positive axons extended to the edge of the lesion, and a few axons followed astrocyte processes into the margins of the lesion site. In contrast to the other pathways, BDA-labeled ascending sensory axons did extend into and arborized extensively within the connective tissue matrix, although the subgroup of ascending axons that are positive for CGRP did not. These results indicate that the connective tissue matrix is permissive for regeneration of some classes of ascending sensory axons but not for other axonal systems.

Inoue, M., T. Hojo, et al. (2003). "The effects of electroacupuncture on peripheral nerve regeneration in rats." Acupunct Med 21(1-2): 9-17.
 This study was designed to examine the effects of electroacupuncture with direct current (DC) on peripheral nerve regeneration. The left sciatic nerve of 55 7-month-old rats was crushed at the thigh. They were ramdomly allocated to four groups: distal cathode DC group (n = 15), distal anode DC group (n = 14), sham operated group (n = 13), and control group (n = 13). In the distal cathode DC group, a cathode electrode was connected to an insulated acupuncture needle inserted at 1 cm distal to the injured site, while an anode electrode was connected to a needle inserted at 1 cm proximal to the lesion. In the distal anode DC group, the anode and the cathode electrode were connected to the needle at 1 cm distal and proximal to the lesion respectively. In the sham operated group, no electrical stimulation was given to the insulated needle inserted at the same site, and in the control group, no treatment was given. Regeneration of the sciatic nerve was evaluated by the number of evoked EMGs recorded at 12 sites in the plantar region, by their latency, and by the weight ratio of the tibialis anterior at four weeks after the crush injury. Regeneration of the peripheral nerve was faster and more accelerated in the distal cathode DC group than in the other groups, while in the distal anode DC group the regeneration was delayed. This result suggested electroacupuncture with cathode distal orientation might be a useful treatment having the advantage of enabling deeper insertion with minimal tissue damage.

Iseda, T., T. Nishio, et al. (2003). "Spontaneous regeneration of the corticospinal tract after transection in young rats: collagen type IV deposition and astrocytic scar in the lesion site are not the cause but the effect of failure of regeneration." J Comp Neurol 464(3): 343-55.
 In young rats the corticospinal tract regenerated after a single transection of the spinal cord with a sharp blade, but regeneration failed if the transection was repeated to make a more traumatic injury. To identify cells and associated molecules that promote or impede regeneration, we compared expression of collagen type IV, glial fibrillary acidic protein (GFAP), and vimentin immunoreactivity (IR) at the lesion sites in combination with anterograde axonal tracing between animals with two types of transection. Axonal regeneration occurred as early as 18 hours after transection; regenerating axons penetrated vessel-like structures with collagen type IV-IR at the lesion site, while reactive astrocytes coexpressing GFAP- and vimentin-IR appeared in the lesioned white matter. In contrast, when regeneration failed astrocytes were absent near the lesion. By 7 days sheet-like structures with collagen type IV-IR and astrocytic scar appeared in the lesioned white matter and persisted until the end of the observation period (31 days). On the basis of their spatiotemporal appearance, collagen type IV-IR sheet-like structures and the astrocytic scar follow, rather than cause, the failure of regeneration. The major sign, and perhaps cause, of failure of axonal regeneration is likely the prolonged disappearance of astrocytes around the lesion site in the early postinjury period.

Isacson, O., L. M. Bjorklund, et al. (2003). "Toward full restoration of synaptic and terminal function of the dopaminergic system in Parkinson's disease by stem cells." Ann Neurol 53 Suppl 3: S135-46; discussion S146-8.
 New therapeutic nonpharmacological methodology in Parkinson's disease (PD) involves cell and synaptic renewal or replacement to restore function of neuronal systems, including the dopaminergic (DA) system. Using fetal DA cell therapy in PD patients and laboratory models, it has been demonstrated that functional motor deficits associated with parkinsonism can be reduced. Similar results have been observed in animal models with stem cell-derived DA neurons. Evidence obtained from transplanted PD patients further shows that the underlying disease process does not destroy transplanted fetal DA cells, although degeneration of the host nigrostriatal system continues. The optimal DA cell regeneration system would reconstitute a normal neuronal network capable of restoring feedback-controlled release of DA in the nigrostriatal system. The success of cell therapy for PD is limited by access to preparation and development of highly specialized dopaminergic neurons found in the A9 and A10 region of the substantia nigra pars compacta as well as the technical and surgical steps associated with the transplantation procedure. Recent laboratory work has focused on using stem cells as a starting point for deriving the optimal DA cells to restore the nigrostriatal system. Ultimately, understanding the cell biological principles necessary for generating functional DA neurons can provide many new avenues for better treatment of patients with PD.

Ishino, T., M. Shirai, et al. (2003). "Identification of genes induced in regenerating Xenopus tadpole tails by using the differential display method." Dev Dyn 226(2): 317-25.
 To identify candidate gene(s) involved in the tail regeneration of Xenopus laevis tadpoles, we used the differential display method to isolate four genes (clones 1, 2, 13a, and 13b) whose expression is induced in regenerating tadpole tails. Among them, clones 13a and 13b were found to encode the Xenopus homologues of the alpha1 chain of type XVIII collagen and neuronal pentraxin I, respectively. Expression of clone 2 and neuronal pentraxin I genes increased dramatically in the blastema 3 days after amputation, whereas that for the clone 1 and type XVIII collagen genes was induced gradually after amputation. In situ hybridization revealed that the neuronal pentraxin I gene is expressed specifically in the regenerating tail epidermis but not in the normal tail epidermis or the most distal margin of the tail blastema, suggesting that it has a tissue-inductive role in tail regeneration. Expression of the four genes was induced in the limb and in the tail blastema, suggesting that they are involved in the regeneration of both organs. Finally, expression of clone 2 and neuronal pentraxin I genes was scarce during embryonic stages in comparison to the tail blastema, suggesting that their main functions are in organ regeneration. Our results demonstrate unique features of spatial and temporal gene expression patterns during Xenopus tadpole tail regeneration.

Islamov, R. R., V. Chintalgattu, et al. (2003). "Differential expression of endothelin receptors in regenerating spinal motor neurons in mice." Brain Res Mol Brain Res 116(1-2): 163-7.
 On day 4 after sciatic nerve crush injury, expression and localization of endothelin receptors ET(A) and ET(B) in the lumbar spinal cord were examined. Immunohistochemical staining with antibodies to ET(A) and ET(B) receptors showed cytoplasmic distribution of ET(A) receptors in motor neurons, whereas ET(B) receptors were localized in the perinuclear region. On the injured side of the lumbar spinal cord, when compared to contralateral, results demonstrated an up-regulation of ET(B) and a down-regulation of ET(A) receptors expression at the level of both mRNA and protein. These results suggest that ET(B) receptors may play a role in the regeneration of axotomized motor neurons.

Islamov, R. R., W. A. Hendricks, et al. (2003). "Effect of 17 beta-estradiol on gene expression in lumbar spinal cord following sciatic nerve crush injury in ovariectomized mice." Brain Res 966(1): 65-75.
 Previously, we observed that estrogen treatment enhances regeneration of the sciatic nerve after crush injury [Brain Res. 943 (2002) 283]. In this research, we studied expression of estrogen receptors and effects of estrogen on gene expression in the lumbar spinal cord, following sciatic nerve crush injury. Using the Atlas Mouse 1.2 Array, changes in the expression of 267 of 1176 genes were registered 4 days after nerve injury. Those genes that exhibited a change in signal intensity ratios of 2-fold or greater were selected as up-regulated (42) or down-regulated (21). In estrogen treated mice, we have observed up-regulation of the genes known to control apoptosis, cell proliferation, and growth, which might account for the positive effects of estrogen on the regeneration of motor neurons. Immunohistochemical staining revealed estrogen receptor-alpha and estrogen receptor-beta localized in the nucleus and cytoplasm of lumbar motor neurons, and in the regenerating neurites of the sciatic nerve. Expression of estrogen receptor-alpha and estrogen receptor-beta mRNA in lumbar spinal cord was shown by traditional RT-PCR. Using real-time quantitative RT-PCR, we demonstrated increased expression of estrogen receptors-alpha and -beta mRNA on the injured side of the lumbar spinal cord. Western blot analysis showed the accumulation of ERs in regenerating sciatic nerve, and revealed a 40% increase of activated ERK1/2 in estrogen treated mice, compared to placebo. Our findings indicate that: (i). axotomized motor neurons increase expression of estrogen receptors-alpha and -beta mRNA, (ii). estrogen mediates the expression of genes which accelerate the growth and maturation of axons, and (iii). estrogen receptors are transported from the perikaryon into regenerating neurites, and estrogen promotes regeneration locally through the non-genomic ERK-activated signaling pathway.

Ito, T., T. Nakamura, et al. (2003). "Biodegradation of polyglycolic acid-collagen composite tubes for nerve guide in the peritoneal cavity." Asaio J 49(4): 417-21.
 The rate of biodegradation of new types of polyglycolic acid (PGA)-collagen composite tubes for nerve regeneration was evaluated in the peritoneal cavity. PGA mesh tubes with a diameter of 2 or 4 mm were coated with collagen solution and dried at room temperature. The tubes were then subjected to dehydrothermal treatment (composite tube). A 2 mm PGA-collagen composite tube filled with collagen sponge was also investigated in this study (sponge tube). Tubes with a length of 15 mm were fixed at the parietal peritoneum of BALB/c mice and excised 2 weeks and 1, 2, and 3 months after the operation. The inner areas of the excised tubes were measured microscopically. Statistical analysis was performed by one way ANOVA and Fisher's PLSD test. Although the inner areas of the 2 and 4 mm composite tubes were not maintained 1 month after the operation (62 +/- 6.8% and 21 +/- 3.8%, respectively), they were well maintained in the sponge tubes (83 +/- 6.4%). The inner areas of the sponge tubes were significantly larger than those of the composite tubes until 2 months after surgery. These results suggest that sponge tubes are more suitable than composite tubes for nerve regeneration in the peritoneal cavity.

Ito, T., T. Nakamura, et al. (2003). "Regeneration of hypogastric nerve using a polyglycolic acid (PGA)-collagen nerve conduit filled with collagen sponge proved electrophysiologically in a canine model." Int J Artif Organs 26(3): 245-51.
 The hypogastric nerve (HGN) is a sympathetic nerve in the peritoneal cavity and controls urinary and seminal functions. In this study, the regeneration of HGN was determined by using a new type of an artificial nerve conduit, polyglycolic acid (PGA)-collagen nerve conduit filled with collagen sponge in two dogs. A PGA-collagen nerve conduit (diameter=2 mm) was interposed in a 10 mm gap of the right HGN. The regeneration of the HGN was evaluated electrophysiologically 8 months after the operation. The intraluminal pressure of spermatic duct and the bladder neck were elevated 80 mmHg and 25 mmHg respectively by the stimulation across the regenerated HGN. The prostate contraction was also elicited. The responses diminished after the excision of the regenerated portion of HGN. These results proved the regeneration of HGN and this nerve conduit will be great help for patients who suffer from urinary and seminal disturbances.

Ito, J. (2003). "[Regeneration of the auditory pathway]." Nippon Rinsho 61(3): 469-74.
 In order to investigate the possibility of the treatment of sensorineural hearing disturbance, experiments were performed using animals. First, the central cochlear pathway in the brain stem to pons was transected in adult rats. Tissue from embryos was transplanted to the lesion site. In 20% of the rats examined, the axons regrew beyond the transected site and regenerated into the denervated side and terminated at the normal targets. The hearing function of animals was also recovered. Those findings contradict the widely held view that the adult mammalian central auditory system cannot be restored following damage. Then, adult rat hippocampus-derived neural stem cells(NSC) were grafted into newborn rat cochlea. Within two to four weeks of grafting to the cochlea, some NSC survived in the cochlear cavity. Some of them had adopted the morphologies and positions of hair cells. This suggests that NSC can adapt to the environment of the cochlea and gives hope for treatment of the damaged cochlea and sensorineural hearing loss.

Itoh, S., I. Yamaguchi, et al. (2003). "Hydroxyapatite-coated tendon chitosan tubes with adsorbed laminin peptides facilitate nerve regeneration in vivo." Brain Res 993(1-2): 111-23.
 On the inner surface of tendon chitosan tubes having a triangular shape and a hydroxyapatite coating (t-chitosan/HAp tube), laminin-1 and laminin peptides (YIGSR, IKVAV) have been adsorbed in order to develop nerve growth conduits. The mechanical property, biocompatibility and efficacy of these tubes for nerve regeneration were examined. Step-1: bridge grafting (15 mm) into the sciatic nerve of Sprague-Dawley (SD) rats was carried out using either t-chitosan or t-chitosan/HAp tubes having either a circular or triangular cross section (N=12 in each group). Specimens were taken after 2-, 4-, 6- and 8-week post-implantation (N=3 in each group) for histology determinations. Step-2: t-chitosan/HAp tubes having a triangular cross section with adsorbed laminin-1, CDPGYIGSR or CSRARKQAASIKVAVSAD, as well as control tubes without pre-adsorption were used for implantation (N=18 in each group). Isografting was also carried out (N=6). Histological evaluation was carried out similarly as in Step-1. Furthermore, evoked muscle and sensory nerve action potentials were recorded, and the percentage of myelinated axon area measured at 10 mm distance of the distal anastomosed site in the experimental, control and isograft groups after 12 weeks (N=6 in each group). The results of histological findings, as well as mechanical properties, suggest that a triangular tube shape with a HAp coating benefits nerve regeneration. The effect of laminin peptides (YIGSR, followed by IKVAV) to enhance the growth of regenerating axons has been found comparable with intact laminin-1. Although histological regeneration in both the YIGSR- and laminin-1-treated t-chitosan/HAp tubes matches the isografts, the functional recovery is however delayed.

Jacobs, W. B. and M. G. Fehlings (2003). "The molecular basis of neural regeneration." Neurosurgery 53(4): 943-48; discussion 948-50.
 THE CENTRAL NERVOUS SYSTEM (CNS) is incapable of meaningful regeneration of lost neurons or axonal and dendritic connections after injury. This often results in permanent and severe loss of neurological function. The CNS regenerative process is unsuccessful for at least three reasons: neurons are highly susceptible to death after CNS injury; the CNS extracellular milieu contains multiple inhibitory factors that make it nonpermissive to growth; and the intrinsic growth capacity of postmitotic neurons is constitutively reduced. However, a number of recent developments in each of these areas is providing insight into the cellular mechanisms involved in CNS regeneration and may eventually lead to the development of therapies capable of effecting successful CNS regeneration.

Jang, W., S. L. Youngentob, et al. (2003). "Globose basal cells are required for reconstitution of olfactory epithelium after methyl bromide lesion." J Comp Neurol 460(1): 123-40.
 Despite a remarkable regenerative capacity, recovery of the mammalian olfactory epithelium can fail in severely injured areas, which subsequently reconstitute as aneuronal respiratory epithelium (metaplasia). We contrasted the cellular response of areas of the rat epithelium that recover as olfactory after methyl bromide lesion with those undergoing respiratory metaplasia in order to identify stem cells that restore lesioned epithelium as olfactory. Ventral olfactory epithelium is at particular risk for metaplasia after lesion and patches of it are rendered acellular by methyl bromide exposure. In contrast, globose basal cells (GBCs, marked by staining with GBC-2) are preserved in surrounding ventral areas and uniformly throughout dorsal epithelium, which consistently and completely recovers as olfactory after lesion. Over the next few days, neurons reappear, but only in those areas in which GBCs are preserved and multiply. In contrast, parts of the epithelium in which GBCs are destroyed are repopulated in part by Bowman's gland cells, which pile up above the basal lamina. Electron microscopy confirms the reciprocity between gland cells and globose basal cells. By 14 days after lesion, the areas that are undergoing metaplasia are repopulated by typical respiratory epithelial cells. As horizontal basal cells are eliminated from all parts of the ventral epithelium, the data suggest that GBC-2(+) cells are ultimately responsible for regenerating olfactory neuroepithelium. In contrast, GLA-13(+) cells may give rise to respiratory metaplastic epithelium where GBCs are eliminated. Thus, we support the idea that a subpopulation of GBCs is the neural stem cell of the olfactory epithelium.

Jasmin, L., A. Boudah, et al. (2003). "Long-term effects of decreased noradrenergic central nervous system innervation on pain behavior and opioid antinociception." J Comp Neurol 460(1): 38-55.
 Here we examine whether a permanent reduction in the noradrenergic (NA) innervation of the spinal cord leads to a chronic decreased nociceptive threshold. NA denervation of rats was achieved by intrathecal injection of dopamine beta-hydroxylase antibodies conjugated to the toxin saporin. A subset of animals also underwent unilateral L5 spinal nerve ligature to induce sustained neuropathic pain behavior. NA fibers and terminals were lost throughout the spinal cord 2 weeks after toxin application and were still absent 12 months later, indicating that regeneration did not occur. There was also a widespread loss of NA terminals in the cerebral cortex, whereas innervation of the hypothalamus and amygdala were close to normal and NA innervation of the brainstem was moderately reduced. There was extensive loss of NA cells in the locus coeruleus and A5 and A7 cell groups. Dopaminergic and serotoninergic innervation was normal. Intracerebroventricular injection of the toxin resulted in additional NA reduction in the hypothalamus, amygdala, and A1 and A2 cell groups. Long-term removal of NA afferents did not affect nociceptive thresholds. Neuropathic animals showed greater mechanical hyperalgesia in the affected hindpaw only during the first 60 days after toxin. Rats lacking NA spinal afferents were less responsive to the antinociceptive effects of morphine, especially in the neuropathic hindpaw, and did not display opioid-dependent stress analgesia. Finally, in the spinal cord of toxin-treated rats, immunoreactivity for substance P was decreased, whereas that of its receptor (NK1) was increased. These animals exhibited antinociception to a low dose of an NK1 receptor antagonist. Our results suggest that NA contributes only modestly to determining the nociceptive threshold and that its antinociceptive effects are closely linked to opioidergic and tachykinergic neurotransmission.

Jazayeri, M., M. R. Ghavanini, et al. (2003). "A study of the sympathetic skin response and sensory nerve action potential after median and ulnar nerve repair." Electromyogr Clin Neurophysiol 43(5): 277-9.
 The purpose of this study was to compare SSR with sensory nerve action potential (SNAP) responses in regeneration of injured peripheral nerves after nerve repair. We studied 10 male patients with a mean age of 26.7 years. All the patients had complete laceration of median or ulnar nerves. The patients were followed up at least for six months. SSR and SNAP assessment were performed every one to two months. Normal hands were used as controls. SSR was positive after 15.8 +/- 9.4 weeks (mean +/- 2 SD) and SNAP after 27.8 +/- 12.9 weeks (mean +/- 2 SD). The difference was statistically significant (P value < 0.001). This can be due to more rapid growth of sympathetic unmyelinated fibers relative to sensory myelinated fibers. This study also shows that recovery of the sudomotor activity following nerve repair is satisfactory in general and SSR can be used as a useful and sensitive method in the evaluation of sudomotor nerve regeneration.

Jean, I. and C. Fressinaud (2003). "Spontaneous central nervous system remyelination is not altered in NFH-lacZ transgenic mice after chemical demyelination." J Neurosci Res 73(1): 54-60.
 Harmonious functioning of the nervous system depends on neuron-glia interactions, particularly between the axons and their myelinating cells, i.e., oligodendrocytes (OL) in the central nervous system (CNS). In human demyelinating diseases such as multiple sclerosis (MS), demyelination may be associated with axonal damage, but alterations of the axonal cytoskeleton, which is composed mainly of neurofilaments (NF) and microtubules, are largely unknown, as are the consequences on remyelination. In a model of demyelination induced by lysophosphatidylcholine (LPC), we have shown that demyelination was correlated with a decrease in NF immunolabelling, and that these axonal abnormalities were reduced by platelet-derived growth factor (PDGF)-enhanced remyelination in adult rats. We have analysed the spontaneous remyelination after LPC stereotaxic injection in the CNS of transgenic NFH-lacZ mice, which present axonal atrophy caused by abnormal distribution of NF, associated with hypermyelination in the PNS, and normal myelin thickness in the CNS. Axonal atrophy in the CNS of NFH-lacZ mice was confirmed, but it was not worsened by demyelination. On the contrary, demyelination induced axonal atrophy in wild-type mice, demonstrating that NF are essential for axonal calibre determination. Moreover, an efficient spontaneous remyelination occurred in NFH-lacZ as well as in wild-type mice, indicating that the NF are not necessary for CNS remyelination. These findings point out that NF modifications observed in MS may not be responsible for the lack of remyelination in this disease.

Jean, I., C. Lavialle, et al. (2003). "Neurotrophin-3 specifically increases mature oligodendrocyte population and enhances remyelination after chemical demyelination of adult rat CNS." Brain Res 972(1-2): 110-8.
 In human central nervous system (CNS) demyelinating diseases, spontaneous remyelination is often incomplete. Therefore, we have tested whether neutrotrophin-3 (NT-3) accelerates CNS myelin repair after a chemically-induced demyelination. One group of adult rats was injected in the corpus callosum (CC) with 1 microl of 1% lysophosphatidylcholine (LPC) and 1 microl of NT-3 (1 microg/microl), and 15 days after injury (D15) remyelination was compared to control rats (receiving 1 microl of LPC+1 microl of vehicle buffer of NT-3). The demyelinated volume decreased by 56% in NT-3-treated rats at D15, and immunohistochemistry showed an increase in mature MBP(+) oligodendrocytes (OL) (+66%) in treated animals (whereas less mature (CNP(+)) OL were unchanged). Since less than 3% axons degenerate in this model, and as astrocytic gliosis was not modified, these data suggest that NT-3 acts directly on cells of the OL lineage to enhance remyelination in vivo.

Jejurikar, S. S. and W. M. Kuzon Jr (2003). "Satellite cell depletion in degenerative skeletal muscle." Apoptosis 8(6): 573-8.
 Adult skeletal muscle has the striking ability to repair and regenerate itself after injury. This would not be possible without satellite cells, a subpopulation of cells existing at the margin of the myofiber. Under most conditions, satellite cells are quiescent, but they are activated in response to trauma, enabling them to guide skeletal muscle regeneration. In degenerative skeletal muscle states, including motor nerve denervation, advanced age, atrophy secondary to deconditioning or immobilization, and Duchenne muscular dystrophy, satellite cell numbers and proliferative potential significantly decrease, contributing to a diminution of skeletal muscle's regenerative capacity and contractility. This review will highlight the fate of satellite cells in several degenerative conditions involving skeletal muscle, and will attempt to gauge the relative contributions of apoptosis, senescence, impaired proliferative potential, and host factors to satellite cell dysfunction.

Jiang, S., J. Wang, et al. (2003). "Enteric glia promote regeneration of transected dorsal root axons into spinal cord of adult rats." Exp Neurol 181(1): 79-83.
 After spinal cord injury axonal regeneration is poor, but may be enhanced by the implantation of olfactory ensheathing glia (OEG). Enteric glia (EG) share many properties of OEG. Transected dorsal root axons normally do not regenerate through the central nervous system myelin into the spinal cord. We tested whether EG, like OEG, could promote regeneration in this paradigm. Three weeks after EG implantation, numerous regenerating dorsal root axons reentered the spinal cord. Ingrowth of dorsal root axons was observed using 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate. Primary sensory afferents invaded laminae 1, 2, and 3, grew through laminae 4 and 5, and reached the dorsal gray commissure. No axonal ingrowth was observed in control animals, indicating that transplanted EG enabled regeneration of the injured dorsal root axons into the adult spinal cord. Thus, EG implantation may be beneficial in promoting axonal growth after central nervous system injury.

Jimeno, D., C. Lillo, et al. (2003). "The degenerative and regenerative processes after the elimination of the proliferative peripheral retina of fish." Exp Neurol 179(2): 210-28.
 We have analyzed the modifications in the tench (Tinca tinca) retina after the complete cryo-elimination of the proliferative growing zone (PGZ), which participates in the continuous growth of the retina throughout the life of the fish. By using immunohistochemistry and electron microscopy we demonstrated that, after the lesion, degenerative and regenerative processes take place in the PGZ, in the ciliary zone, and in the transition zone located between the PGZ and the central retina. After 120 days postlesion, the PGZ was completely regenerated and its composition was similar to that of the control animals. Numerous proliferative PCNA-positive cells reappeared and new ganglion cells were formed. In the transition zone and the central retina numerous proliferative PCNA-positive cells also appeared. These are arranged, on occasion, as columnar units from the inner to the outer nuclear layer where the rod precursors and the progenitor cells, respectively, were located. The Muller cells, closely associated with these columnar units, appeared to use them as guides to migration during the regenerative process. Notably, modifications occurred in the ciliary zone, whose cells acquired similar characteristics to the PGZ cells. The ciliary zone cells, the Muller cells, the rod precursors, and the proliferative cells located in the inner nuclear layer appear to participate actively in the regeneration of the PGZ.

Jones, L. L., D. Sajed, et al. (2003). "Axonal regeneration through regions of chondroitin sulfate proteoglycan deposition after spinal cord injury: a balance of permissiveness and inhibition." J Neurosci 23(28): 9276-88.
 Increased expression of certain extracellular matrix (ECM) molecules after CNS injury is believed to restrict axonal regeneration. The chondroitin sulfate proteoglycans (CSPGs) are one such class of ECM molecules that inhibit neurite outgrowth in vitro and are upregulated after CNS injury. We examined growth responses of several classes of axons to this inhibitory environment in the presence of a cellular fibroblast bridge in a spinal cord lesion site and after a growth factor stimulus at the lesion site (fibroblasts genetically modified to secrete NGF). Immunohistochemical analysis showed dense labeling of the CSPGs NG2, brevican, neurocan, versican, and phosphacan at the host-lesion interface after spinal cord injury (SCI). Furthermore, robust expression of NG2, and to a lesser extent versican, was also observed throughout grafts of control and NGF-secreting fibroblasts. Despite this inhibitory milieu, several axonal classes penetrated control fibroblast grafts, including dorsal column sensory, rubrospinal, and nociceptive axons. Axon growth was amplified more in the presence of NGF-secreting grafts. Confocal microscopy demonstrated that axon growth was, paradoxically, preferentially associated with NG2-rich substrates in both graft types. NG2 expression also increased after sciatic nerve injury, wherein axons successfully regenerate. Cellular sources of NG2 in SCI and peripheral nerve lesion sites included Schwann cells and endothelial cells. Notably, these same cellular sources in lesion sites produced the cell adhesion molecules L1 and laminin, and these molecules all colocalized. Thus, axons grow along substrates coexpressing both inhibitory and permissive molecules, suggesting that regeneration is successful when local permissive signals balance and exceed inhibitory signals.

Jones, L. L., R. U. Margolis, et al. (2003). "The chondroitin sulfate proteoglycans neurocan, brevican, phosphacan, and versican are differentially regulated following spinal cord injury." Exp Neurol 182(2): 399-411.
 Chondroitin sulfate proteoglycans (CSPGs) are extracellular matrix (ECM) molecules that are widely expressed throughout the developing and adult CNS. In vitro studies demonstrate their potential to restrict neurite outgrowth, and it is believed that CSPGs also inhibit axonal regeneration after CNS injury in vivo. Previous studies demonstrated that CSPGs are generally upregulated after spinal cord injury, and more recent reports have begun to identify individual proteoglycans that may play dominant roles in limiting axonal regeneration. The current study systematically examined the extended deposition patterns after CNS injury of four putatively inhibitory CSPGs that have not been extensively investigated previously in vivo: neurocan, brevican, phosphacan, and versican. After spinal cord injury, neurocan, brevican, and versican immunolabeling increased within days in injured spinal cord parenchyma surrounding the lesion site and peaked at 2 weeks. Neurocan and versican were persistently elevated for 4 weeks postinjury, and brevican expression persisted for at least 2 months. On the other hand, phosphacan immunolabeling decreased in the same region immediately following injury but later recovered and then peaked after 2 months. Combined glial fibrillary acidic protein (GFAP) immunohistochemistry and in situ hybridization demonstrated that GFAP astrocytes constituted a source of neurocan production after spinal cord injury. Thus, the production of several CSPG family members is differentially affected by spinal cord injury, overall establishing a CSPG-rich matrix that persists for up to 2 months following injury. Optimization of strategies to reduce CSPG expression to enhance regeneration may need to target several different family members over an extended period following injury.

Jordanova, A., P. De Jonghe, et al. (2003). "Mutations in the neurofilament light chain gene (NEFL) cause early onset severe Charcot-Marie-Tooth disease." Brain 126(Pt 3): 590-7.
 Neurofilament light chain polypeptide (NEFL) is one of the most abundant cytoskeletal components of the neuron. Mutations in the NEFL gene were recently reported as a cause for autosomal dominant Charcot-Marie-Tooth type 2E (CMT2E) linked to chromosome 8p21. In order to investigate the frequency and phenotypic consequences of NEFL mutations, we screened 323 patients with CMT or related peripheral neuropathies. We detected six disease associated missense mutations and one 3-bp in-frame deletion clustered in functionally defined domains of the NEFL protein. Patients have an early onset and often a severe clinical phenotype. Electrophysiological examination shows moderately to severely slowed nerve conduction velocities. We report the first nerve biopsy of a CMT patient with a de novo missense mutation in NEFL, and found an axonal pathology with axonal regeneration clusters and onion bulb formations. Our findings provide further evidence that the clinical variation observed in CMT depends on the gene mutated and the specific type of mutation, and we also suggest that NEFL mutations need to be considered in the molecular evaluation of patients with sporadic or dominantly inherited CMT.

Jubran, M. and J. Widenfalk (2003). "Repair of peripheral nerve transections with fibrin sealant containing neurotrophic factors." Exp Neurol 181(2): 204-12.
 Peripheral nerve injury is often followed by incomplete recovery of function and sometimes associated with neuropathic pain. There is, therefore, need for therapies which improve the speed of recovery and the final functional outcome after peripheral nerve injuries. In addition, neuropathic pain is not easily dealt with clinically and should preferably be eliminated. Neurotrophic factors have well-documented abilities to support neuron survival and stimulate neurite outgrowth, making them excellent candidates for use in repairing injured nerves. We investigated the possible beneficial effects of repairing the transected rat sciatic nerve by local application of a fibrin sealant containing nerve growth factor (NGF), glial cell line-derived neurotrophic factor (GDNF), or acidic fibroblast growth factor (aFGF). Fibrin sealant was used in conjunction with sutures. Evaluation of motor and sensory function, autotomy, and histological parameters was carried out from 1 to 12 weeks after injury. We demonstrate that NGF cotreatment decreased the occurance of autotomy, suggesting a reduction of neuropathic pain, and improved the performance in motor and sensory tests. In addition, the number of regenerating motoneurons was significantly increased after NGF administration. GDNF increased the speed of sensory recovery, but also markedly increased autotomy, indicating an increased degree of neuropathic pain. aFGF did not alter the outcome of the motor or sensory tests. Fibrin sealant could easily be used in conjunction with sutures to deliver neurotrophic substances locally to the damaged nerve and to enhance recovery of nerve function.

Jurynec, M. J., C. P. Riley, et al. (2003). "TIGR is upregulated in the chronic glial scar in response to central nervous system injury and inhibits neurite outgrowth." Mol Cell Neurosci 23(1): 69-80.
 Reactive astrocytes respond to central nervous system (CNS) injury and disease by elaborating a glial scar that is inhibitory to axonal regeneration. To identify genes that may be involved in the astrocytic response to injury, we used differential display polymerase chain reaction and an in vivo model of the CNS glial scar. Expression of the trabecular meshwork inducible glucocorticoid response (TIGR) gene was increased in gliotic tissue compared with the uninjured cerebral cortex. Increased TIGR expression by reactive astrocytes was confirmed by in situ hybridization, quantitative reverse transcriptase-polymerase chain reaction, immunoblot analysis, and immunohistochemistry. Although mutations of the TIGR gene have been implicated in glaucoma, a function for TIGR has not been reported. Since TIGR is secreted, we assessed a possible role in inhibition of neuronal regeneration with an in vitro bioassay and found that this protein is a potent inhibitor of neurite outgrowth. Thus, TIGR is a newly identified component of the CNS glial scar that is likely to contribute to neuronal regenerative failure characteristic of the mammalian CNS.

Kaas, J. H. and C. E. Collins (2003). "Anatomic and functional reorganization of somatosensory cortex in mature primates after peripheral nerve and spinal cord injury." Adv Neurol 93: 87-95.
 
Kakinoki, R., R. Ikeguchi, et al. (2003). "Treatment of painful peripheral neuromas by vein implantation." Int Orthop 27(1): 60-4.
 Firstly, we designed a vein-implantation model using the rat femoral nerve and vein to study the morphometric changes in nerve endings inserted into venous lumina. By 4 weeks, nerve fibers had extended from the nerve stump into the lumen of the vein and along the endothelium of the vein. After 8 weeks, the lengths and number of nerve fibres extending into the vein lumen began to decrease. At 12 weeks, the nerve ending had developed a hemispherical shape. In none of the experiments was a neuroma formed. Secondly, we treated ten neuromas in ten patients by the vein-implantation method. We obtained excellent results in seven patients.

Kamijo, Y., J. Koyama, et al. (2003). "Regenerative process of the facial nerve: rate of regeneration of fibers and their bifurcations." Neurosci Res 46(2): 135-43.
 After the main trunk of the mouse facial nerve was injured by crushing, a fiber tracing method was used to quantify the facial motor neurons that extended regenerating nerve fibers to the specific site of the facial nerve branch. The total number of motor neurons retrogradely labeled with a fluorescent tracer, Fluoro-Gold (FG), were 0 on postsurgical days (PSDs) 1 and 2, 75+/-25 on PSD3, 264+/-21 on PSD4, 378+/-19 on PSD6, 428+/-19 on PSD8, 491+/-13 on PSD12 and 532+/-15 on PSD16. Assuming that the FG-positive neurons (535+/-11) of the control mice represent 100%, the FG-labeled neurons accounted for 0, 14, 49, 71, 80, 92 and 99% on the corresponding days. Two different fluorescent tracers were applied to the different facial nerve branches 16 days after facial nerve injuries. Double-labeled neurons were consistently found in the nerve-crushed facial nucleus (3.2%), and their number increased in the nerve-transected facial nucleus (12.2%). The present study indicates that the regenerating facial nerve consists of heterogeneous nerve fibers with varying growth rates and that excessive axonal branching occurs more frequently in the nerve-transected than in the nerve-crushed injuries.

Kanemaru, S., T. Nakamura, et al. (2003). "Recurrent laryngeal nerve regeneration by tissue engineering." Ann Otol Rhinol Laryngol 112(6): 492-8.
 The recurrent laryngeal nerve (RLN) does not regenerate well after it has been cut, and no current surgical methods achieve functional regeneration. Here, we evaluate the functional regeneration of the RLN after reconstruction using a biodegradable nerve conduit or an autologous nerve graft. The nerve conduit was made of a polyglycolic acid (PGA) tube coated with collagen. A 10-mm gap in the resected nerve was bridged by a PGA tube in 6 adult beagle dogs (group 1) and by an autologous nerve graft in 3 dogs (group 2). Fiberscopic observation revealed functional regeneration of the RLN in 4 of the 6 dogs in group 1. No regeneration of the RLN was observed in any dog in group 2. We also tested for axonal transport, and measured the compound muscle action potential. The RLN can be functionally regenerated with a PGA tube, which may act as a scaffold for the growth of regenerating axons.

Kapur, T. A. and M. S. Shoichet (2003). "Chemically-bound nerve growth factor for neural tissue engineering applications." J Biomater Sci Polym Ed 14(4): 383-94.
 In order to promote regeneration after spinal cord injury, growth factors have been applied in vivo to rescue ailing neurons and provide a path finding signal for regenerating neurites. We previously demonstrated that soluble growth factor concentration gradients can guide axons over long distances, but this model is inherently limited to in vitro applications. To translate the use of growth factor gradients to an implantible device for in vivo studies, we developed a photochemical method to bind nerve growth factor (NGF) to microporous poly(2-hydroxyethylmethacrylate) (PHEMA) gels and tested bioactivity in vitro. A cell adhesive photoreactive poly(allylamine) (PAA) was synthesized and characterized. This photoreactive PAA was applied to the surface of the PHEMA gels to provide both a cell adhesive layer and a photoreactive handle for further NGF immobilization. Using a direct ELISA technique, the amount of NGF immobilized on the surface of PHEMA after UV exposure was determined to be 5.65 +/- 0.82 ng/cm2 or 3.4% of the originally applied NGF. A cell-based assay was performed to determine the bioactivity of the immobilized NGF. Using pheochromocytoma (PC-12) cells, 30 +/- 7% of the cell population responded to bound NGF, a response statistically similar to that of cells cultured on collagen in the presence of 40 ng/ml soluble NGF of 39 +/- 12%. These results demonstrate that PHEMA with photochemically bound NGF is bioactive. This photochemical technique may be useful to spatially control the amount of NGF bound to PHEMA using light and thus build a stable concentration gradient.

Kato, R., S. Kiryu-Seo, et al. (2003). "Cavernous nerve injury elicits GAP-43 mRNA expression but not regeneration of injured pelvic ganglion neurons." Brain Res 986(1-2): 166-73.
 Recovery of erectile dysfunction after cavernous nerve injury takes a long period. To elucidate this mechanism, unilateral cavernous nerve of male rat was cut, and the expression level of a nerve regeneration marker, the growth associated protein-43 (GAP-43) mRNA was evaluated by in situ hybridization and RT-PCR. While GAP-43 mRNA expression was transiently increased in the injured neurons of the major pelvic ganglion (MPG) at 7 days after nerve injury, continuous increase of GAP-43 mRNA was observed in the contralateral MPG from 7 days to 6 months after the nerve injury. Histochemical double-labeling studies for either neuronal NOS (nNOS) or tyrosine hydroxylase (TH) and the GAP-43 mRNA expression demonstrated that in injured MPG the transient up-regulation of GAP-43 mRNA was mainly seen in nNOS negative and/or TH positive neurons, suggesting non-parasympathetic post-ganglionic neurons, and also demonstrated that in contralateral MPG GAP-43 mRNA positive neurons were gradually increased in nNOS positive but TH negative neurons, suggesting parasympathetic post-ganglionic neurons. When a retrograde tracer Fluorogold (FG) was injected into the penile crus 7 days before histological experiments, FG-positive neurons were, if any, hardly seen in nNOS-positive neurons of the injured MPG for at least 6 months, whereas numerous FG-positive cells were seen in nNOS-positive neurons of the contralateral MPG. These results suggest that post-ganglionic projecting neurons of the intact side, which express increased GAP-43 mRNA, would be most likely to contribute to the recovery of the erectile function after unilateral cavernous nerve injury possibly by a plastic change such as nerve sprouting.

Kawamoto, K., S. Ishimoto, et al. (2003). "Math1 gene transfer generates new cochlear hair cells in mature guinea pigs in vivo." J Neurosci 23(11): 4395-400.
 Hair cell loss in the mammalian cochlea is irreversible and results in permanent hearing loss. Math1, the basic helix-loop-helix transcription factor homolog of the Drosophila atonal gene, is a positive regulator of hair cell differentiation during cochlear development. Developing hair cells express Math1, and nonsensory cells do not. We set out to determine the outcome of overexpression of Math1 in nonsensory cells of the cochlea on the phenotype of these cells. We demonstrate that in vivo inoculation of adenovirus with the Math1 gene insert into the endolymph of the mature guinea pig cochlea results in Math1 overexpression in nonsensory cochlear cells, as evident from the presence of Math1 protein in supporting cells of the organ of Corti and in adjacent nonsensory epithelial cells. Math1 overexpression leads to the appearance of immature hair cells in the organ of Corti and new hair cells adjacent to the organ of Corti in the interdental cell, inner sulcus, and Hensen cell regions. Axons are extended from the bundle of auditory nerve toward some of the new hair cells, suggesting that the new cells attract auditory neurons. We conclude that nonsensory cells in the mature cochlea retain the competence to generate new hair cells after overexpression of Math1 in vivo and that Math1 is necessary and sufficient to direct hair cell differentiation in these mature nonsensory cells.

Kawasaki, T., N. Oka, et al. (2003). "Oct6, a transcription factor controlling myelination, is a marker for active nerve regeneration in peripheral neuropathies." Acta Neuropathol (Berl) 105(3): 203-8.
 Three transcription factors, Krox20 (EGR2), Oct6 (SCIP/Tst1) and Sox10, are considered necessary for transition from the nonmyelinating to the myelinating stage of Schwann cell development. We immunohistochemically studied Oct6 expression in peripheral nerve specimens from 25 patients with various diseases including Charcot-Marie-Tooth disease type 1A (CMT1A). Oct6 was present in cytoplasm of Schwann cells associated with normal-appearing myelinated nerve fibers, but not in nuclei. Expression was seen in nuclei of Schwann cells in the early phase of acute axonal degeneration; nuclear expression peaked at the regenerative stage. Schwann cells forming "onion bulbs" expressed Oct6 in chronic inflammatory demyelinating polyneuropathy (CIDP), but showed minimal expression in CMT1A, reflecting their proliferative activity in CIDP. Nerves showing chronic axonal loss had no expression. Oct6, then, may be a marker for dedifferentiation of adult Schwann cells and active nerve regeneration.

Keilhoff, G., H. Fansa, et al. (2003). "Nitric oxide synthase, an essential factor in peripheral nerve regeneration." Cell Mol Biol (Noisy-le-grand) 49(6): 885-97.
 Nitric oxide (NO) exerts both, pro-apoptotic and anti-apoptotic actions and appears to be acritical factor inneuronal degenerative and regenerative processes. NO is synthesized from L-arginine by NO synthase occurring in three isoforms of (neuronal, nNOS; endothelial, eNOS; inducible, iNOS). In a mice sciatic nerve model the regenerative outcome was assessed when the endogenous NO supply was deficient by knocking out the respective NOS isoform and compared to that of wild type mice after nerve transection. In nNOS knock-out mice a delay in regeneration, preceded by slowedWallerian degeneration and a disturbed pruning of uncontrolled sprouts, was observed. This was associated with a delayed recovery of sensory and motor function. Additionally, deficiency of nNOS led after nerve cut to a substantial loss of small and medium-sized dorsal root ganglia neurons, spinal cord interneurons and, to a lesser extent, spinal cord motor neurons. A lack of iNOS resulted in a delayed Wallerian degeneration and impaired regenerative outcome without consequences for neuronal survival. A lack of eNOS was well tolerated, although a delay in nerve revascularization was observed. Thus, after peripheral nerve lesion, regular NOS activity is essential for cell survival and recovery with reference to the nNOS isoform.

Keilhoff, G., F. Stang, et al. (2003). "Bio-compatibility of type I/III collagen matrix for peripheral nerve reconstruction." Biomaterials 24(16): 2779-87.
 Nerve gaps are usually bridged by autografts. With improving technical methods biocompatible conduits may become an alternative graft to reconstruct nerves. Non-neural conduits fail to support regeneration over larger gaps due to lacking viable Schwann cells. Thus, tissue engineering of nerves is focusing on implantation of viable Schwann cells into suitable scaffolds. In this study, we tested collagen type I/III tubes as a potential nerve guiding matrix. Revascularization, foreign body reaction, biodegradation and Schwann cell settlement were evaluated by immunocytochemistry, light, fluorescence and scanning electron microscopy, after different implantation times. The conduits were completely revascularized between day 5 and 7 post-operatively and well integrated into the host tissue. Host response was characterized by a moderate invasion of ED1/ED2-positive macrophages. Biodegradation of the tubes was slowly enough to maintain a stable support structure for extended regeneration processes. Implanted Schwann cells adhered, survived and proliferated on the inner surface of the conduits and were able to form nerve guiding columns of Bungner. From this results, we conclude that collagen-type I/III can serve as template to design "living" nerve conduits, which may be able to ensure nerve regeneration through extended nerve gaps.

Kerber, G., R. Streif, et al. (2003). "Neuregulin-1 isoforms are differentially expressed in the intact and regenerating adult rat nervous system." J Mol Neurosci 21(2): 149-65.
 Our knowledge on Neuregulin-1 (Nrg-1) during development of the nervous system is increasing rapidly, but little is known about Nrg-1-ErbB signaling in the adult brain. Nrg-1 is involved in determination, proliferation, differentiation, and migration of neurons and glial cells in the developing brain. In the peripheral nervous system, Nrg-1 signaling is required for Schwann cell differentiation and myelination, and establishment of neuromuscular junctions (NMJs). Multiple alternative splicing of Nrg-1 was shown, but correlation of its structural and functional diversity was rarely addressed. Therefore, we investigated the expression of Nrg-1 isoforms in the rat brain and brain-derived cell types, and their involvement in regeneration of the adult brain, using immunohistochemistry, in situ hybridization, and semiquantitative RT-PCR. We found expression of at least 12 distinct Nrg-1 isoforms in the brain and altered expression of several isoforms in the facial motor nucleus after peripheral transection of the seventh cranial nerve. An upregulation of Nrg-1 type-I mRNA, probably type- I-alpha, was observed in reactive astrocytes of the facial nucleus 1 d postaxotomy. Nrg-1 type-III and the splice variants beta1 and beta5 are dramatically downregulated in axotomized motoneurons, which lack contact to their target tissue. Baseline expression levels were reestablished when the first axons reached the facial muscles and reformed NMJs. Nrg-1-beta1 and -beta5 might act in maintenance of NMJs. The splice variants beta2 and beta4 display an initial downregulation of mRNA levels, followed by an increase during the period of axon remyelination. Thus, Nrg- 1-beta2 and -beta4 might be involved in myelination.

Kerns, J. M., N. Danielsen, et al. (2003). "A comparison of peripheral nerve regeneration in acellular muscle and nerve autografts." Scand J Plast Reconstr Surg Hand Surg 37(4): 193-200.
 Regeneration of the rat sciatic nerve through acellular muscle and nerve autografts was evaluated 6-28 days postoperatively by the sensory pinch test, immunocytochemical staining for neurofilaments, and light and electron microscopy. Data points generated by the pinch test were plotted against postoperative time periods and by the use of regression analysis the initial delay period for muscle grafts was determined to 10.3 days. This value was similar to that previously published for acellular nerve grafts (9.5 days), but significantly longer than that for fresh nerve grafts (3.6 days). The calculated regeneration rate (slope of the regression line) for muscle grafts (1.8 mm/day) did not differ significantly (p > 0.05) from that calculated for acellular nerve grafts (2.1 mm/day) or for fresh nerve grafts (1.5 mm/day). The front of regenerating axons shown by axonal neurofilament staining confirmed the pinch test results. Both types of acellular grafts were repopulated with host non-neuronal cells and the muscle graft contained occasional ectopic muscle fibres. Remnants of graft basal laminae were evident at the ultrastructural level. These results indicate the suitability of either acellular muscle or nerve grafts for nerve repair despite their prolonged initial delay periods compared with conventional fresh nerve grafts.

Kerns, J. M., S. Shott, et al. (2003). "Effects of IGF-I gene therapy on the injured rat pudendal nerve." Int Urogynecol J Pelvic Floor Dysfunct 14(1): 2-7; discussion 8.
 Injured nerves and their motor units may undergo enhanced recovery when exposed to recombinant human insulin-like growth factor-I (rhIGF-I). The external anal sphincter muscle in the female rat was denervated to model incontinence. The treatment-group muscle was injected with rhIGF-1 plasmid, whereas in the control group the plasmid lacked the cDNA insert and the normal group received neither surgery nor treatment. Electromyography data at 56 days post surgery indicated more reinnervation without fibrillation potentials in the treatment group (2 of 6) than in the control group (0 of 6). The histology of the regenerated axons in the pudendal nerve distal to the crush site also suggested an improved recovery in the treatment group. The number of motor neurons retrogradely labeled with horseradish peroxidase was decreased by 50% following pudendal nerve crush in both experimental groups compared to the normal group. We conclude from these preliminary results that rhIGF-I gene therapy may improve the distal recovery of structure and function.

Khodr, B., J. Howard, et al. (2003). "Effect of short-term and long-term antioxidant therapy on primary and secondary ageing neurovascular processes." J Gerontol A Biol Sci Med Sci 58(8): 698-708.
 Previous studies from our laboratory demonstrated an age-related functional decline in sensory neurones and their modulation of microvascular blood flow (primary ageing processes) that correlated with a deficiency in tissue repair (a secondary ageing process). We also raised the notion of a possible role for free radicals in these age-related changes. The aim of this study was to investigate the impact of antioxidant therapy on modulating sensory neurovascular function and tissue repair with age. Twenty-four-month-old Sprague-Dawley rats were treated with vitamin E for short-term (40 mg/kg, i.p., every other day for 2 weeks) or long-term (for 12 months in advance, 10 g/kg, incorporated in food). These treated rats were assessed for the effectiveness of treatment and tested for their sensory neurovascular function, repair of full-thickness burn, and recovery from hyperalgesia following nerve injury. The results indicate that both short- and long-term vitamin E treatments are effective in improving sensory neurovascular function and in reducing the time required for complete wound closure of full-thickness burn injury. Short-term vitamin E treatment was more effective in protecting against the development of hyperalgesia following nerve injury. An initial increase in wound size and in hyperalgesia was observed in the treated animals, and could reflect possible side effects of the antioxidant therapy and support the importance of free radicals in early stages of the repair process. The data, overall, support the notion that oxidative damage contributes to both primary and secondary ageing processes.

Kieseier, B. C. and H. P. Hartung (2003). "Multiple paradigm shifts in multiple sclerosis." Curr Opin Neurol 16(3): 247-52.
 PURPOSE OF REVIEW: The present article reviews the currently ongoing scientific debate of our changing views on the pathogenesis of multiple sclerosis and the therapeutic strategies currently available for multiple sclerosis. RECENT FINDINGS: The most important observations include that (a) axonal loss accounts for permanent disability in multiple sclerosis, (b) remyelination should be possible in theory but fails for unknown reasons in the multiple sclerosis lesion, (c) inflammation can be beneficial, (d) treatment should be initiated early, and (e) immunosuppressive strategies exhibit beneficial effects in progressive forms of the disease. SUMMARY: Our current understanding of the immunopathogenesis of multiple sclerosis has changed in the past. Whereas demyelination was originally thought to be relevant for the lasting neurological deficit, it is nowadays commonly accepted that the extent of axonal loss dictates the degree of permanent clinical disability. How axonal damage can be prevented remains elusive. The interaction between the myelinating cell and the neuron gains increasing attention, however the evolving knowledge has not yet yielded new treatment concepts. Hence for the time being, it seems prudent to make optimal use of current approved therapies. Recent trials underlined the need for early initiation of treatment with immunomodulatory drugs. The superiority of one of the interferons is still a matter of debate, and a conclusive answer cannot be given at present. Finally, with mitoxantrone we have a drug at hand which can be used in progressive forms of multiple sclerosis, especially when other disease modifying drugs are not or no longer effective.

Kikuchi, K., A. Kishino, et al. (2003). "In vitro and in vivo characterization of a novel semaphorin 3A inhibitor, SM-216289 or xanthofulvin." J Biol Chem 278(44): 42985-91.
 SM-216289 (xanthofulvin) isolated from the fermentation broth of a fungal strain, Penicillium sp. SPF-3059, was identified as a strong semaphorin 3A (Sema3A) inhibitor. Sema3A-induced growth cone collapse of dorsal root ganglion neurons in vitro was completely abolished in the presence of SM-216289 at levels less than 2 mum (IC50 = 0.16 mum). When dorsal root ganglion explants were co-cultured with Sema3A-producing COS7 cells in a collagen gel matrix, SM-216289 enabled neurites to grow toward the COS7 cells. SM-216289 diminished the binding of Sema3A to its receptor neuropilin-1 in vitro, suggesting a direct interference of receptor-ligand association. Moreover, our data suggest that SM-216289 interacted with Sema3A directly and blocked the binding of Sema3A to its receptor. We examined the efficacy of SM-216289 in vivo using a rat olfactory nerve axotomy model, in which strong Sema3A induction has been reported around regenerating axons. The regeneration of olfactory nerves was significantly accelerated by a local administration of SM-216289 in the lesion site, suggesting the involvement of Sema3A in neural regeneration as an inhibitory factor. SM-216289 is an excellent molecular probe to investigate the function of Sema3A, in vitro and in vivo, and may be useful for the treatment of traumatic neural injuries.

Kim, D. H., Y. J. Cho, et al. (2003). "Surgical outcomes of 111 spinal accessory nerve injuries." Neurosurgery 53(5): 1106-12; discussion 1102-3.
 OBJECTIVE: Iatrogenic injury to the spinal accessory nerve is not uncommon during neck surgery involving the posterior cervical triangle, because its superficial course here makes it susceptible. We review injury mechanisms, operative techniques, and surgical outcomes of 111 surgical repairs of the spinal accessory nerve. METHODS: This retrospective study examines clinical and surgical experience with spinal accessory nerve injuries at the Louisiana State University Health Sciences Center during a period of 23 years (1978-2000). Surgery was performed on the basis of anatomic and electrophysiological findings at the time of operation. Patients were followed up for an average of 25.6 months. RESULTS: The most frequent injury mechanism was iatrogenic (103 patients, 93%), and 82 (80%) of these injuries involved lymph node biopsies. Eight injuries were caused by stretch (five patients) and laceration (three patients). The most common procedures were graft repairs in 58 patients. End-to-end repair was used in 26 patients and neurolysis in 19 patients if the nerve was found in continuity with intraoperative electrical evidence of regeneration. Five neurotizations, two burials into muscle, and one removal of ligature material were also performed. More than 95% of patients treated by neurolysis supported by positive nerve action potential recordings improved to Grade 4 or higher. Of 84 patients with lesions repaired by graft or suture, 65 patients (77%) recovered to Grade 3 or higher. The average graft length was 1.5 inches. CONCLUSION: Surgical exploration and repair of spinal accessory nerve injuries is difficult. With perseverance, however, these patients with complete or severe deficits achieved favorable functional outcomes through operative exploration and repair.

Kim, J. E., I. E. Bonilla, et al. (2003). "Nogo-C is sufficient to delay nerve regeneration." Mol Cell Neurosci 23(3): 451-9.
 Axonal regeneration succeeds in the peripheral but not central nervous system of adult mammals. Peripheral clearance of myelin coupled with selective CNS expression of axon growth inhibitors, such as Nogo, may account for this reparative disparity. To assess the sufficiency of Nogo for limiting axonal regeneration, we generated transgenic mice expressing Nogo-C in peripheral Schwann cells. Nogo-C includes the panisoform inhibitory Nogo-66 domain, but not a second Nogo-A-specific inhibitory domain, allowing a selective consideration of the Nogo-66 region. The oct-6::nogo-c transgenic mice regenerate axons less rapidly than do wild-type mice after mid-thigh sciatic nerve crush. The delayed axonal regeneration is associated with a decreased recovery rate for motor function after sciatic nerve injury. Thus, expression of the Nogo-66 domain by otherwise permissive myelinating cells is sufficient to hinder axonal reextension after trauma.

Kim, D. H., Y. J. Cho, et al. (2003). "Outcomes of surgery in 1019 brachial plexus lesions treated at Louisiana State University Health Sciences Center." J Neurosurg 98(5): 1005-16.
 OBJECT: Outcomes of 1019 brachial plexus lesions in patients who underwent surgery at Louisiana State University Health Sciences Center during a 30-year period are reviewed in this paper to provide management guidelines. METHODS: Causes of brachial plexus lesions included 509 stretches/contusions (50%), 161 plexus tumors (16%), 160 thoracic outlet syndromes (TOSs, 16%), 118 gunshot wounds (12%), and 71 lacerations (7%). Many features of clinical presentation, including prior treatment, patient's neurological status, results of electrophysiological studies, intraoperative findings, and postoperative level of function, were studied. The minimum follow-up period was 18 months and the mean follow-up period was 42 months. Repairs were best for injuries located at the C-5, C-6, and C-7 levels, the upper and middle trunk, the lateral cord to the musculocutaneous nerve, and the median and posterior cords to the axillary and radial nerves. Conversely, results were poor for injuries at the C-8 and T-1 levels, and for lower trunk and medial cord lesions, with the exception of injuries of the medial cord to the median nerve. Outcomes were most favorable when patients were carefully evaluated and selected for surgery, although variables such as lesion type, location, and severity, as well as time since injury also affected outcome. This was true also of TOSs and tumors arising from the plexus, especially if they had not been surgically treated previously. CONCLUSIONS: Surgical exploration and repair of brachial plexus lesions is technically feasible and favorable outcomes can be achieved if patients are thoroughly evaluated and appropriately selected.

Kim, J. E., S. Li, et al. (2003). "Axon regeneration in young adult mice lacking Nogo-A/B." Neuron 38(2): 187-99.
 After injury, axons of the adult mammalian brain and spinal cord exhibit little regeneration. It has been suggested that axon growth inhibitors, such as myelin-derived Nogo, prevent CNS axon repair. To investigate this hypothesis, we analyzed mice with a nogo mutation that eliminates Nogo-A/B expression. These mice are viable and exhibit normal locomotion. Corticospinal tract tracing reveals no abnormality in uninjured nogo-A/B(-/-) mice. After spinal cord injury, corticospinal axons of young adult nogo-A/B(-/-) mice sprout extensively rostral to a transection. Numerous fibers regenerate into distal cord segments of nogo-A/B(-/-) mice. Recovery of locomotor function is improved in these mice. Thus, Nogo-A plays a role in restricting axonal sprouting in the young adult CNS after injury.

Kimura, H., M. Kawatani, et al. (2003). "Effects of pituitary adenylate cyclase-activating polypeptide on facial nerve recovery in the Guinea pig." Laryngoscope 113(6): 1000-6.
 OBJECTIVES/HYPOTHESIS: Pituitary adenylate cyclase-activating polypeptide (PACAP) has neurotrophic effects of neural regeneration and gives protection to the nervous system. We investigated whether PACAP had a neurotrophic effect on peripheral motoneurons and whether PACAP could facilitate glial cell line-derived neurotrophic factor (GDNF), a neurotrophin, in nerve regeneration. The presence and distribution of PACAP receptors following facial nerve transection were also investigated. STUDY DESIGN: Animal experiment. METHODS: Unilateral transection of the facial nerve was performed in male Hartley guinea pigs, and PACAP was injected at the site. Saline was substituted as a control. Compound muscle action potentials were recorded to measure the changes of latency. Glial cell line-derived neurotrophic factor (GDNF) content in facial target muscle was measured using enzyme-linked immunosorbent assay. The regenerating site was taken for histological studies. RESULTS: Pituitary adenylate cyclase-activating polypeptide hastened the appearance of compound muscle action potentials and shortened the latency. Pituitary adenylate cyclase-activating polypeptide increased and prolonged the nerve transection-induced GDNF increase in the facial muscles. The number of myelinated fibers at 1 to 4 weeks after the transection was increased. PAC1 receptor or VPAC1 receptor or both were identified in the injury area at 2 to 4 weeks. CONCLUSIONS: Pituitary adenylate cyclase-activating polypeptide facilitated the recovery of latency of compound muscle action potentials or the number of myelinated axons, or both. Pituitary adenylate cyclase-activating polypeptide prolonged the GDNF levels in target organs. These data indicated that PACAP promoted regeneration of the facial nerve.

King, C. E., A. Wallace, et al. (2003). "Transient up-regulation of retinal EphA3 and EphA5, but not ephrin-A2, coincides with re-establishment of a topographic map during optic nerve regeneration in goldfish." Exp Neurol 183(2): 593-9.
 Eph tyrosine kinase receptors and their ligands, the ephrins, play a key role in the establishment of retinotectal topography during development. Tectal up-regulation of ephrin-A2 in goldfish, coincident with the reestablishment of a retinotectal map, suggests a similar role during optic nerve regeneration. Here we report a complementary study of EphA3, EphA5 and ephrin-A2 expression in the retina. EphA3 and EphA5 are transiently up-regulated as ascending naso-temporal gradients, whereas ephrin-A2 remains uniform. The expression profiles differ from those in developing chick and mouse, suggesting that different combinations of retinal Eph receptors and ligands can generate topographic guidance information.

King, C. T., L. D. Deyrup, et al. (2003). "Effects of gustatory nerve transection and regeneration on quinine-stimulated Fos-like immunoreactivity in the parabrachial nucleus of the rat." J Comp Neurol 465(2): 296-308.
 The distribution of quinine-stimulated Fos-like immunoreactivity (FLI) in several subdivisions of the parabrachial nucleus (PBN) known to be responsive to gustatory stimulation was examined in rats in which the chorda tympani nerve (CT) and/or glossopharyngeal nerve (GL) was transected (Experiment 1) and in rats in which the GL was transected with regeneration promoted or prevented (Experiment 2). We confirmed previous findings in the literature by demonstrating that rats intraorally infused with 3 mM quinine showed a robust population of FLI in the waist area and the external lateral (EL) and external medial (EM) subdivisions of the PBN (Yamamoto et al. [1994] Physiol Behav 56:1197-1202; Travers et al., [ 1999] Am J Physiol 277:R384-R394). In the waist area, only GL transection significantly decreased the number of FLI-neurons elicited by intraoral infusion of quinine compared with water-stimulated controls. In the external subdivisions neither neurotomy affected the number of FLI-neurons. The effect of GL transection in the waist area was enduring for rats in which the GL did not regenerate (up to 94 days), but regeneration of the GL after 52 days restored quinine-stimulated FLI to control values. In these same GL-transected animals, there were parallel decreases in the number of gapes elicited by intraoral quinine stimulation that recovered, but only subsequent to regeneration of the GL. These data provide support for the role of the waist area in the brainstem processing that underlies oromotor rejection behaviors and also help substantiate the hypothesis that the CT and GL are relatively specialized with regard to function. Moreover, when the quinine-induced pattern of neural activity in the second central gustatory relay, as assessed by FLI, is substantially altered by the loss of peripheral gustatory input from the GL, it can be restored upon regeneration of the nerve.

King, V. R., M. Henseler, et al. (2003). "Mats made from fibronectin support oriented growth of axons in the damaged spinal cord of the adult rat." Exp Neurol 182(2): 383-98.
 A variety of biological as well as synthetic implants have been used to attempt to promote regeneration into the damaged spinal cord. We have implanted mats made from fibronectin (FN) into the damaged spinal cord to determine their effectiveness as a substrate for regeneration of axons. These mats contain oriented pores and can take up and release growth factors. Lesion cavities 1 mm in width and depth and 2 mm in length were created on one side of the spinal cord of adult rats. FN mats containing neurotrophins or saline were placed into the lesion. Mats were well integrated into surrounding tissue and showed robust well-oriented growth of calcitonin gene-related peptide, substance P, GABAergic, cholinergic, glutamatergic, and noradrenergic axons into FN mats. Transganglionic tracing using cholera toxin B indicated large-diameter primary afferents had grown into FN implants. Schwann cells had also infiltrated FN mats. Electron microscopy confirmed the presence of axons within implants sites, with most axons either ensheathed or myelinated by Schwann cells. Mats incubated in brain-derived neurotrophic factor and neurotrophin-3 showed significantly more neurofilament-positive and glutamatergic fibers compared to saline- and nerve growth factor-incubated mats, while mats incubated with nerve growth factor showed more calcitonin gene-related peptide-positive axons. In contrast, neurotrophin treatment had no effect on PGP 9.5-positive axons. In addition, in some animals with neurotrophin-3-incubated mats, cholera toxin B-labelled fibers had grown from the mat into adjoining intact areas of spinal cord. The results indicate that FN mats provide a substrate that is permissive for robust oriented axonal growth in the damaged spinal cord, and that this growth is supported by Schwann cells.

Kirby, B. P. and J. N. Rawlins (2003). "The role of the septo-hippocampal cholinergic projection in T-maze rewarded alternation." Behav Brain Res 143(1): 41-8.
 Administration of 192IgG-saporin, a cholinergic neurotoxin, to the medial septum destroys the cell bodies from which the septo-hippocampal cholinergic projection originates, leading to reductions in both hippocampal acetylcholinesterase (AChE) and choline acetyltransferase (ChAT). Despite reports that 192IgG-saporin-induced cholinergic loss leads to post-operative impairments in acquisition and performance of spatial memory tasks, a number of other reports have described intact spatial memory performance following these lesions. Factors that might account for these different outcomes include variations in toxin injection sites or volumes, and post-operative testing at times that might permit regeneration of damaged neuronal processes. We, therefore, assessed the effects of intraseptal microinjection of 192IgG-saporin, in rats, on the post-operative retention of pre-operatively acquired discrete-trial rewarded alternation in the T-maze. This design allowed us to assess the effects of the lesion 7 days post-surgery, at which point, at best, incomplete neuronal regeneration would have been expected to have occurred. The lesion led to a profound loss of hippocampal AChE staining, and a clear inflammatory response, as assessed by proliferation of OX42-stained macrophages in the medial septum and diagonal band nuclei, but there was no impairment in spatial working memory.

Kishino, A. and C. Nakayama (2003). "Enhancement of BDNF and activated-ERK immunoreactivity in spinal motor neurons after peripheral administration of BDNF." Brain Res 964(1): 56-66.
 Brain-derived neurotrophic factor (BDNF) shows neurotrophic effects on adult motor neurons when given systemically, But it is unknown whether systemically administered BDNF is transported to central cell bodies to affect them directly. Here we used immunohistochemistry to investigate the transport of peripherally injected BDNF to spinal motor neurons and the subsequent activation of a signaling pathway. We first injected BDNF into the flexor digitorum brevis (FDB) and analyzed the motor nucleus that projects to the FDB for BDNF immunoreactivity (BDNF-ir) and phosphorylated extracellular signal-regulated kinase (ERK) 1/2 immunoreactivity (pERK1/2-ir). Both immunoreactivities were observed in the motor neuron cell bodies. Next, BDNF was injected subcutaneously (s.c.) into rats with a unilaterally axotomized sciatic nerve. pERK1/2-ir was detected in motor neurons of the lesioned side. BDNF-ir and pERK1/2-ir were also observed on the unlesioned side when a high dose of BDNF was injected. Therefore, we examined BDNF-ir and pERK1/2-ir after injecting BDNF s.c. into normal rats. Both immunoreactivities were observed in motor nuclei on both sides. Finally, we examined pERK1/2-ir after a lower dose of BDNF was injected, which prevents the decrease in choline acetyl transferase that occurs in the motor neuron upon axotomy. Spinal motor nuclei contained a few cell bodies with pERK1/2-ir. These findings represent the first direct evidence that subcutaneously injected BDNF is transported to motor neurons and that it activates a signaling pathway in the spinal cord and exhibits neurotrophic effects in vivo.

Kiwaki, T., F. Umehara, et al. (2003). "The clinical and pathological features of peripheral neuropathy accompanied with HTLV-I associated myelopathy." J Neurol Sci 206(1): 17-21.
 We describe the clinical and pathological studies in HTLV-I associated myelopathy (HAM)/tropical spastic paraparesis (TSP) patients with peripheral neuropathy as proven by sural nerve biopsy. Sural nerve pathology in HAM/TSP patients revealed that the most common type of pathologic change is a combination of both demyelination and remyelination and axonal degeneration and regeneration, and this change is modified by the complications. The pathologic changes were correlated with neither the duration of disease nor human T lymphotropic virus type I (HTLV-I) proviral load. This study suggests that peripheral nerves could be involved in HAM/TSP.

Kobayashi, T., K. Kihara, et al. (2003). "Spontaneous regeneration of the seriously injured sympathetic pathway projecting to the prostate over a long period in the dog." BJU Int 91(9): 868-72.
 OBJECTIVES: To explore the spontaneous regeneration, over a long period, of the seriously injured sympathetic pathway controlling the prostate. MATERIALS AND METHODS: The hypogastric nerve (HGN), which is part of the sympathetic pathway from the spinal cord to the prostate, was partly removed over half of its length on both sides in six dogs. Four years after surgery the responses of the prostate to electrical stimulation of the lumbar splanchnic nerve (LSN) or the HGN proximal to the site removed, were assessed. RESULTS: In six dogs, 10 of the 17 LSNs (second to fourth) and four of the 10 HGNs stimulated elicited prostatic contraction. The pathways via the ipsilateral HGN and/or the contralateral HGN from the LSNs to the prostate were identified as having regenerated in four of six dogs. CONCLUSION: These results indicate that the sympathetic pathways via the HGN to the canine prostate can regenerate spontaneously over a long period after serious injury, and that their cross-innervation system can also be repaired.

Kokaia, Z. and O. Lindvall (2003). "Neurogenesis after ischaemic brain insults." Curr Opin Neurobiol 13(1): 127-32.
 Evidence for neuronal self-repair following insults to the adult brain has been scarce until very recently. Ischaemic insults have now been shown to trigger neurogenesis from neural stem cells or progenitor cells located in the dentate subgranular zone, the subventricular zone lining the lateral ventricle, and the posterior periventricle adjacent to the hippocampus. New neurons migrate to the granule cell layer or to the damaged CA1 region and striatum, where they express morphological markers characteristic of those neurons that have died. Some evidence indicates that these neurons can re-establish connections and contribute to functional recovery.

Kolb, B. and J. Cioe (2003). "Recovery from early cortical damage in rats. IX. Differential behavioral and anatomical effects of temporal cortex lesions at different ages of neural maturation." Behav Brain Res 144(1-2): 67-76.
 Rats were given lesions of the temporal association cortex on postnatal day 4 or 10, or in adulthood. Ninety days later they were trained on two visual tasks (visual-spatial navigation; horizontal-vertical stripes discrimination). Lesion animals were compared behaviorally and neuroanatomically to littermate sham control rats. The day 4 lesions produced a larger deficit in the navigation task than day 10 or adult lesions. There were no deficits in the discrimination task. Analysis of the brains showed that the day 4 lesions produced a smaller brain and thinner cortex than day 10 lesions. The day 10 lesions produced hypertrophy in the dendritic arborization of pyramidal cells in parietal cortex. The results are consistent with the general findings that perinatal cortical injury in rats produces more severe behavioral and morphological effects than similar lesions in the second week of life and that cortical lesions around day 10 lead to an increase in cortical synaptogenesis.

Kolosova, L. I., A. D. Nozdrachev, et al. (2003). "[Effect of central axotomy of sensory neurones on the mechanoreception recovery in initial regeneration of the injured sciatic nerve in rats]." Ross Fiziol Zh Im I M Sechenova 89(5): 579-84.
 No apparent effect of lumbar dorsal rhizotomy performed simultaneously with the peripheral nerve injury, has been revealed on the triggering of regeneration of sensitive nerve fibers. Re-innervation of the foot skin by either decentralized regeneration of nerve fibers or those sustaining their connections with the central nervous system (CNS), has been shown to start 30 days after surgery. Using the recording of impulse activity of a single nerve fibre, the mechanical thresholds of decentralized regenerating receptors were found to be significantly higher as compared to the thresholds of the regenerating receptors sustaining their connections with the CNS. The findings suggest that afferent nerve fibers and mechanical receptors formed on the periphery, continue functioning after decentralization and sustain their regenerative capacity after injury. However, in marked contrast, the decrease in sensitivity of regenerating receptors is more pronounced.

Komiyama, T., Y. Nakao, et al. (2003). "A novel technique to isolate adult Schwann cells for an artificial nerve conduit." J Neurosci Methods 122(2): 195-200.
 The use of an artificial nerve conduit containing viable Schwann cells (SCs) is one of the most promising approaches to repair nerve injuries. Obtaining a large number of viable SCs in a short period is demanded for the clinical use of this technique. However, the previous methods using mitogens are not clinically acceptable, and other methods that do not require mitogens, failed to isolate adult SCs effectively or required a long period of time. In this study, we have developed a novel technique to isolate SCs from adult rat peripheral nerves for an artificial nerve conduit without mitogens, which has produced a total number of 1.21 x 10(5) cells per mg, with an average purity of 93.0+/-0.58% at 21 days in vitro. The Bottenstein-Sato (BS) medium used in this study, had originally been developed for oligodendrocyte culture, but here it is shown to have an effect on SC proliferation and survival. By changing fetal bovine serum (FBS) concentrations from 0 to 10% serially, SCs could be isolated maximally from the predegenerated nerves while suppressing fibroblast overgrowth. The combination of this technique and the altered medium promoted the migration and proliferation of SCs selectively by utilizing the supporting cells of SCs instead of discarding them by changing the culture dishes and media.

Koshima, I., Y. Nanba, et al. (2003). "Deep peroneal nerve transfer for established plantar sensory loss." J Reconstr Microsurg 19(7): 451-4.
 Patients with established or irreversible plantar sensory loss often have normal sensation on the dorsal aspect of the foot, due to an intact deep peroneal nerve. A new method of deep peroneal nerve transfer is proposed for repair of plantar sensory loss caused by extensive nerve gaps or high-level lesions of the posterior tibial nerve. Two cases in which this technique was used are described. The surgical technique is relatively easy, with a short operating time, rapid nerve regeneration after surgery, accurate sensory recovery, and minimal donor-site morbidity with sensory loss only on the first web space of the foot.

Koshimune, M., K. Takamatsu, et al. (2003). "Creating bioabsorbable Schwann cell coated conduits through tissue engineering." Biomed Mater Eng 13(3): 223-9.
 Enormous effort has been devoted to the generation of a synthetic guidance conduit for nerve repair instead of utilizing autograft. Several studies show neural guidance conduit is more effective when coated with Schwann cells. In this study, we synthesized bioabsorbable conduit consist of L-lactide and epsilon-caprolactone which was useful clinically and examined adhesion of Schwann cells to bioabsorbable conduits. In vivo studies were done in which these polymer conduits coated with Schwann cells were implanted across a 12 mm gap in the rat sciatic nerve. Silicone conduits were implanted across the same gap as control. At 12 weeks, axonal regeneration was observed in the midconduit region of these polymer conduits and was not in control. This study assesses the feasibility of a tissue engineering approach to constructing bioabsorbable conduits coated with Schwann cells.

Kotulska, K., J. Lewin-Kowalik, et al. (2003). "Bcl-2 deficiency deprives peripheral nerves of neurotrophic activity against injured optic nerve." J Neurosci Res 73(6): 846-52.
 Optic nerve injury leads to retinal ganglion cell apoptosis, thus preventing fiber regeneration. Peripheral nerve grafts are known to promote survival and regeneration in injured adult mammalian central nervous system, including optic nerve, but the mechanisms of their activity remain unclear. It is likely that they attenuate the apoptotic cascade triggered by axotomy in retinal ganglion cells. The aim of this work was to examine the role of the antiapoptotic gene bcl-2 in the optic nerve regeneration induced by such grafts. Experiments were carried out on bcl-2-deficient and wild-type mice. We have reported previously that predegeneration markedly enhances neurotrophic activity of peripheral nerve grafts, so we applied both predegenerated and non-predegenerated implants to the transected optic nerves. We studied the neurotrophic effects of bcl-2-deficient grafts on wild-type and bcl-2 knock-out optic nerves, as well as wild-type grafts on both strains of mouse optic nerves. After application of fluorescent dye to the end of the graft, we counted the stained retinal ganglion cells. Predegenerated wild-type grafts promoted survival and outgrowth of retinal ganglion cells axons in both types of mice. By contrast, non-predegenerated and predegenerated bcl-2-deficient grafts induced little or no regeneration in the optic nerves. These results indicate that the lack of bcl-2 gene does not deprive retinal ganglion cells of their regenerative potential. At the same time, we found that bcl-2 knock-out dispossesses peripheral nerves of their neurotrophic activity.

Koussoulakos, S., L. H. Margaritis, et al. (2003). "Quantitative estimation of HRP-labeled sensory and motor neurons during nerve-dependent and nerve-independent periods of urodele limb regeneration." Izv Akad Nauk Ser Biol(4): 405-15.
 The relationship between urodele regeneration and possible regeneration in mammalian prospects is hard to evidence, but the idea of possible regeneration of neural elements in people is an area of potential clinical importance that is under investigation. One of the great challenges of the future is to understand enough about the basic biology of animal regeneration and to use it for the betterment of the mankind. It is well established that the initial stages of urodele limb regeneration depend on the presence of intact nerve fibres connected to their cell bodies. The nerve fibres severed at the limb amputation level, regrow and invade the blastema, providing blastema cells with indispensable factors. These factors are elaborated within the neuron perikarya and transported via their axons to the blastema. Numerous studies have been so far performed and have elucidated the quantitative relationships between nerve fibres and limb regeneration. However, there are no reports dealing with the individual nerve cells at work. The aim of the present investigation was to analyse the quantitative participation and qualitative distinction of nerve cells innervating regenerating parts of the urodele limb and their possible interrelationship with the nerve-dependent and nerve-independent periods of regeneration. The cells under study are housed in the dorsal ganglia (sensory neurons) and in the ventral aspect of the spinal cord grey matter (motor neurons). As a means of visualizing the direct implication of these neurons during various regeneration periods, the enzyme horseradish peroxidase was chosen. A total of 34 animals were used, 21 experimental and 13 controls, in order to study labeled nerve cell fluctuations. The results are summarized as follows: (a) The first nerve cells incorporating HRP within 5 days post amputation are found in the dorsal ganglia. Motor neurons in the grey matter are labeled within 7 days. (b) The number of labeled perikarya increases during the nerve-dependent regeneration period (0-21 dpa). The percentage of implicated sensory neurons exceeds that found in the control series. (c) During the next, nerve-independent period, the number of participating labeled neurons decreases gradually. Such fluctuations in the number of labeled neurons might represent the metabolic status of these cells in their effort to provide the blastema cells with the factors needed at the appropriate time. The current findings support previous observations that the periods of dependence and independence of urodele limb regeneration from the integrated control of brachial nerves reflect changes in the metabolism of individual sensory and motor neurons.

Kozorovitskiy, Y. and E. Gould (2003). "Adult neurogenesis: a mechanism for brain repair?" J Clin Exp Neuropsychol 25(5): 721-32.
 It is now generally accepted that new neurons are added to the adult mammalian brain. This raises the possibility that naturally occurring neurogenesis may be useful for repairing the damaged adult brain. Indeed, several studies have shown that damage to the adult brain can stimulate reparative neurogenesis. However, the production of new neurons is only one of several steps necessary to restore damaged neural circuits to their original state. Studies carried out on intact animals have identified several conditions that affect the production and survival of new neurons in adult brains. This review considers the evidence for compensatory neurogenesis in the adult mammalian brain with a view toward applying information from the undamaged brain to studies of regeneration.

Kudlachev, A. V. and V. A. Otellin (2003). "[Perspectives for the clinical application of the results of current neuromorphological studies of the optical nerve lesions and atrophies]." Morfologiia 123(3): 94-102.
 The review presents the analysis of advances in fundamental studies of the optic nerve regeneration and degeneration mechanisms. It is concluded that current experimental morphological investigations of mechanisms of pathogenesis and sanogenesis of the optic nerve atrophies are developing in several major directions. These include: 1) development of techniques for restoration of the integrity of the optic nerve stem, including those that use peripheral nerve grafts; 2) development of measures to decrease the retinal ganglionic cell (RGC) response to injury and to stimulate RGC repair processes; 3) search for the ways for axon growth stimulation in lesioned and undamaged RGC; 4) usage of RGC apoptosis inhibitors; 5) grafting of embryonic retina. Results of the experimental studies are evaluated in the context of their possible application in clinical practice.

Kuntzer, T., M. Dunand, et al. (2003). "Phenotypic expression of a Pro 87 to Leu mutation in the connexin 32 gene in a large Swiss family with Charcot-Marie-Tooth neuropathy." J Neurol Sci 207(1-2): 77-86.
 BACKGROUND: The clinical manifestations of CMTX have been well described but the natural history has not yet been studied in detail. We studied phenotype variability in a family with a Pro 87 to Leu mutation of the connexin 32 (Cx32) gene. METHODS: A total of 32 family members, of which 19 patients were affected, underwent clinical, electrophysiological, and genetic studies. RESULTS: Onset was in the second decade. Clinical features were similar in both sexes when quantitative scores were compared, but more males had a steppage gait and skeletal deformities. All adult patients had a predominant involvement of the thenar muscles. The median values of nerve conduction velocities (NCVs) were not statistically different in men and in women. The correlation coefficients were low between motor NCVs within the same extremities, indicating nonuniform slowing between nerves, the ulnar nerve being the least affected. When disability was rated, a strong correlation was seen in male patients between severity of motor axonal loss and duration of the disease. The main pathological features were axonal loss, clusters of regenerating fibers and paranodal demyelination, the hallmark of a Schwann cell pathology. CONCLUSIONS: Our data support the hypothesis that clinical disability in CMTX is caused by loss of large myelinated axons in men. Furthermore, this study shows that the nerves are not uniformly affected in terms of axonal loss. Preventing axonal degeneration and promoting axonal regeneration in the most affected nerves might be the best therapeutic approaches to ameliorate disability in CMTX.

Kurihara, H., H. Shinohara, et al. (2003). "Neurotrophins in cultured cells from periodontal tissues." J Periodontol 74(1): 76-84.
 We review the basic functions of neurotrophins and their receptors and discuss the expression and functions of neurotrophins and their specific receptors based on recent data using cultured cells from human periodontal tissues. Neurotrophins, nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin-3 (NT-3) play crucial roles in the differentiation and survival of neural cells. Neurotrophins activate 2 different receptor classes: the tropomyosin-related kinase (Trk) family of receptor tyrosine kinases (TrkA, TrkB, and TrkC) and the p75 receptor, a member of the tumor necrosis factor receptor superfamily. Neurotrophins regulate both cell death and cell survival through activations of Trk receptors and/or p75 neurotrophin receptor. It has been reported that neurotrophins are also produced from non-neuronal cells, such as leukocytes, osteoblasts, or fibroblasts, and act in many other ways on non-neuronal cells. Neurotrophin expression during bone fracture healing is especially interesting, and neurotrophins are now implicated in hard tissue regeneration. It is well known that neurotrophins and their receptors are expressed in tooth development. Recent studies have found that neurotrophins and Trk receptors are expressed in mouse osteoblastic cell lines. Human periodontal ligament cells, human gingival fibroblasts, and human gingival keratinocytes expressed mRNA for NGF and TrkA. The secretion of bioactive NGF peptides from human periodontal ligament cells and human gingival keratinocytes was confirmed by bioassay using PC12 cells (rat adrenal pheochromocytoma cells). The expression of NGF and TrkA.mRNA was regulated by interleukin (IL)-1beta. NGF increased DNA synthesis and expressions of mRNA for bone-related proteins, alkaline phosphatase, and osteopontin in human periodontal ligament cells. Neurotrophins and Trk receptors expressed in human periodontal tissue may contribute to regeneration as well as innervation of periodontal tissue through local autocrine and paracrine pathways. Recent data suggest that some functions of neurotrophins and Trk receptors relate to periodontal disease and periodontal tissue regeneration. However, in vivo studies will be required to clarify the roles of neurotrophins and their receptors, including p75, in periodontal disease and periodontal tissue regeneration.

Kvist, M., N. Danielsen, et al. (2003). "Effects of FK506 on regeneration and macrophages in injured rat sciatic nerve." J Peripher Nerv Syst 8(4): 251-9.
 Effects of FK506 [5.0 mg/kg body weight (BW), subcutaneous, daily] on nerve regeneration and presence of macrophages in lesioned rat sciatic nerves were studied. Models of autologous nerve graft or a nerve crush lesion were used and regeneration was evaluated by immunocytochemistry (also used to detect ED1/ED2 macrophages) and sensory pinch reflex test, respectively. Treatment with FK506 did not increase regeneration distance or regeneration rate in the autologous nerve grafts. However, regeneration distances after nerve crush were significantly longer following treatment with FK506. The number of macrophages (ED1/ED2) in nerve grafts increased over time, but treatment with FK506 had limited effects only in the presence of ED2 macrophages. Present and previously published studies may imply that there is a time-related and type-of-injury-related profile of FK506's pro-regenerative effect.

Lambert, C., A. M. Landau, et al. (2003). "Fas-beyond death: a regenerative role for Fas in the nervous system." Apoptosis 8(6): 551-62.
 Fas (CD95, APO-1), a member of the TNF superfamily, is a prototypical "death receptor" which transduces apoptotic signals in a variety of cell types. However, cell death is not the only possible outcome of Fas signalling. Fas engagement by Fas Ligand can also trigger proliferation or differentiation, promote tumour progression and angiogenesis, and induce cytokine secretion and integrin expression. Recently, we have reported that Fas engagement induces a potent regenerative response in sensory neurons in vitro, and enhances peripheral nerve regeneration in vivo. In contrast, other types of neurons, notably motoneurons, are acutely sensitive to Fas-induced apoptosis. Here, we review the literature on non-apoptotic Fas signalling pathways, and discuss the potential roles, molecular mechanisms, and regulators of Fas signalling in the nervous system.

Lambiase, A., L. Manni, et al. (2003). "Clinical application of nerve growth factor on human corneal ulcer." Arch Ital Biol 141(2-3): 141-8.
 Nerve growth factor (NGF) is a neurotrophic and immunomodulatory factor contributing of the control of cutaneous morphogenesis, wound healing and inflammatory responses. Following the evidence that topical administration of NGF leads to healing of human corneal ulcers, we investigate the therapeutic action of NGF on immune and/or autoimmune cutaneous ulcers. We found that 1-10 micrograms of highly purified murine NGF dissolved in 50 microliters of physiological solution and topically applied to skin ulcer leads, after 4-12 weeks of daily treatment (depending on the size and depth of the lesion) to complete healing of the ulcer. Thus, NGF was able to promote complete repair in human skin and corneal ulcers which were poorly or non-responsive to conventional topical and systemic treatments. No side effects were observed and a follow up after 4 months showed no signs of relapse. These findings indicate that failure of cutaneous tissues to produce sufficient amounts of NGF might represent a prominent mechanism implicated in the clinical manifestation of ocular ulcers.

Lancelotta, M. P., R. N. Sheth, et al. (2003). "Severity and duration of hyperalgesia in rat varies with type of nerve lesion." Neurosurgery 53(5): 1200-8; discussion 1208-9.
 OBJECTIVE: To learn how lesions with differing capacity for nerve regeneration affect the severity and duration of hyperalgesia in an animal model of neuropathic pain. METHODS: Three groups of rats were studied: 1). L5 nerve root crush (favorable for regeneration); 2). L5 root ligation and section; and 3). sham-operated group. An experimenter who did not know the rats' groups tested the animals for hyperalgesia to mechanical and cold stimuli. RESULTS: Measures of adverseness of mechanical and cooling stimuli for the crush group and ligation/cut groups were significantly higher than for the sham-operated group (P < 0.001 for both) for the first 30 days after lesioning. By 40 days, the crush group recovered from mechanical hyperalgesia, whereas the ligation/cut group continued to have significant hyperalgesia. At this time, both lesion groups displayed hyperalgesia to the cooling stimulus (P < 0.001), but the hyperalgesia in the ligation/cut group was significantly greater (P < 0.01). No recovery from cooling hyperalgesia was evident during the 54-day period of observation. Histological studies of the sciatic nerve indicated higher numbers of regenerating fibers in the crush group compared with the ligation/cut group. CONCLUSION: This study demonstrates that axotomy, regardless of how it is induced, produces hyperalgesia to both mechanical and cold stimuli. However, the lesion that favors regeneration is associated with earlier signs of recovery from mechanical hyperalgesia and less severe signs of cooling hyperalgesia. The data support the hypothesis that inputs from the injured afferents play an ongoing role in neuropathic pain from nerve injury. Nerve ligation induces more severe and more sustained behavioral signs of pain than nerve crush.

Landi, F., L. Aloe, et al. (2003). "Topical treatment of pressure ulcers with nerve growth factor: a randomized clinical trial." Ann Intern Med 139(8): 635-41.
 BACKGROUND: The prevalence of pressure ulcers of the foot is a major health care problem in frail elderly patients. A pressure sore dramatically increases the cost of medical and nursing care, and effective treatment has always been an essential nursing concern. Management options for pressure ulcers include local wound care; surgical repair; and, more recently, topical application of growth factors. OBJECTIVE: To examine the effects of topical treatment with nerve growth factor in patients with severe, noninfected pressure ulcers of the foot. DESIGN: Randomized, double-blind, placebo-controlled trial. SETTING: Teaching nursing home of Catholic University of the Sacred Heart, Italy. PATIENTS: 36 persons with pressure ulcers of the foot. INTERVENTION: 18 patients received nerve growth factor treatment, and 18 patients received only conventional topical treatment. MEASUREMENTS: The course of the ulcers during follow-up was evaluated by tracing the perimeter of the wound onto sterile, transparent block paper and determining the stage. RESULTS: At baseline, the treatment and control groups did not differ across demographic variables, clinical characteristics, and functional measures. The mean area (+/-SD) of the ulcers was 1012 +/- 633 mm2 in the treatment group and 1012 +/- 655 mm2 in the control group (P > 0.2). The average reduction in pressure ulcer area at 6 weeks was statistically significantly greater in the treatment group than in the control group (738 +/- 393 mm2 vs. 485 +/- 384 mm2; P = 0.034). CONCLUSION: Topical application of nerve growth factor may be an effective therapy for patients with severe pressure ulcers.

Lanzetta, M., A. Gal, et al. (2003). "Effect of FK506 and basic fibroblast growth factor on nerve regeneration using a polytetrafluoroethylene chamber for nerve repair." Int Surg 88(1): 47-51.
 Peripheral nerve repair can be accomplished by using a polytetrafluoroethylene tubular chamber to guide nerve healing and regeneration. In this study, we delivered basic fibroblast growth factor (bFGF) into the chamber for sciatic nerve repair in rats. In addition, the animals were given systemically 1 mg/kg/day FK506 (tacrolimus), a potent immunosuppressant with neurotrophic properties. Nerve regeneration was evaluated by means of a nociceptive test and a grasping test starting 2 weeks postoperatively. Animals that received bFGF and FK506 showed a significantly faster recovery from injury than did the control group. Morphometric analysis at 3 months showed no difference between the two groups in total number of axonal fibers, fiber diameter, fiber density, and myelin:axon ratio. We conclude that the combination of bFGF and low dose FK506 enhances nerve healing in this animal model by accelerating early regrowth but has no effect on the final outcome.

Lee, A. C., V. M. Yu, et al. (2003). "Controlled release of nerve growth factor enhances sciatic nerve regeneration." Exp Neurol 184(1): 295-303.
 Based on previous studies demonstrating the potential of growth factors to enhance peripheral nerve regeneration, we developed a novel growth factor delivery system to provide sustained delivery of nerve growth factor (NGF). This delivery system uses heparin to immobilize NGF and slow its diffusion from a fibrin matrix. This system has been previously shown to enhance neurite outgrowth in vitro, and in this study, we evaluated the ability of this delivery system to enhance nerve regeneration through conduits. We tested the effect of controlled NGF delivery on peripheral nerve regeneration in a 13-mm rat sciatic nerve defect. The heparin-containing delivery system was studied in combination with three doses of NGF (5, 20, or 50 ng/mL) and the results were compared with positive controls (isografts) and negative controls (fibrin alone, NGF alone, and empty conduits). Nerves were harvested at 6 weeks postoperatively for histomorphometric analysis. Axonal regeneration in the delivery system groups revealed a marked dose-dependent effect. The total number of nerve fibers at both the mid-conduit level and in the distal nerve showed no statistical difference for NGF doses at 20 and 50 ng/mL from the isograft (positive control). The results of this study demonstrate that the incorporation of a novel delivery system providing controlled release of growth factors enhances peripheral nerve regeneration and represents a significant contribution toward enhancing nerve regeneration across short nerve gaps.

Lehnert, M., W. I. Steudel, et al. (2003). "Histochemical alterations of re-innervated rat extensor digitorum longus muscle after end-to-end or graft repair: a comparative histomorphological study." J Anat 203(1): 21-9.
 Changes in the histochemical profile of 43 rat extensor digitorum longus muscles undergoing de-innervation and re-innervation were recorded. Assessment of fibre type composition and muscle fibre cross-sectional area was performed at 15, 30, 90 and 180 days post operative (p.o.) after either primary end-to-end repair or autologous graft repair of the common peroneal nerve (n = 5 per time point and type of repair). The size and histochemical profile of single muscle fibres were analysed by computer-assisted quantification on the basis of their myofibrillar ATPase (pH 4.3) and succinate dehydrogenase (SDH) activities in serial, whole-muscle cross-sections. Accordingly, four muscle-fibre types could be functionally identified: (1) slow oxidative (SO, type I); (2) fast-oxidative glycolytic (FOG, type IIA); (3) fast glycolytic (FG, type IIB); and (4) succinate dehydrogenase intermediate (SDH-INT). At 15 days following end-to-end repair, the SDH-INT muscle fibre type was observed. By contrast, 15 days following graft repair, no changes in fibre type composition were observed (vs. control). At 30 days p.o. in the group that received end-to-end repair, type SDH-INT reached its maximum and was significantly higher than in the group that underwent graft repair. At 90 days p.o., the amount of SDH-INT fibres declined after end-to-end repair, but it was still significantly higher than in the group treated with a nerve graft. The increase of the SDH-INT fibre type was mirrored by a proportional disappearance of FG and FOG fibres. These changes were time-dependent, not reversible at 180 days p.o and largely blunted after nerve graft. Muscle-fibre size decreased at 15 and 30 days after both types of nerve repair. This decrease was transient and reversible within 90 days p.o. These findings reflect the fact that the reorganization of the histochemical profile in re-innervated muscles is both time dependent and long lasting. The degree of this reorganization is significantly higher after end-to-end repair than after graft repair.

Leis, A. A., J. A. Lancon, et al. (2003). "Retrograde regeneration following neurotmesis of the ulnar nerve." Muscle Nerve 28(4): 512-4.
 A 41-year-old woman experienced a gunshot wound to the forearm with neurotmesis of the ulnar nerve. Surgery 9 months later revealed a neuroma-in-continuity in the midforearm. Intraoperative nerve stimulation failed to elicit direct nerve responses or motor responses from the first dorsal interosseous (FDI) and abductor digiti minimi (ADM) muscles. However, neurotonic discharges in response to mechanical irritation of the neuroma were recorded in the FDI, but not the ADM. Surprisingly, after resecting the ulnar nerve distal to the neuroma, neurotonic discharges were still elicited in the FDI following perturbation of the neuroma. Moreover, neurotonic discharges were elicited during ulnar nerve resection 2 cm proximal to the neuroma. No anastomoses or anomalous branches were noted. The findings suggest that regenerating fibers did not reach the FDI through the distal nerve segment. Rather, we speculate that nerve fibers regenerating at random, or impeded by scar tissue, contacted the proximal nerve portion, at which point growth became polarized in a retrograde direction. Retrograde regeneration may have proceeded to a branch point in the forearm (possibly an undetected anomalous branch or fibrous adhesion), where growth of regenerating fibers extended outward into surrounding damaged tissue planes before redirecting distally to reach the FDI.

Leszczynski, J. K. and K. A. Esser (2003). "The MEF2 site is necessary for induction of the myosin light chain 2 slow promoter in overloaded regenerating plantaris muscle." Life Sci 73(25): 3265-76.
 Functional overload (OV) of the rat plantaris muscle results in a fast to slow change in muscle phenotype with induction of the slow contractile protein genes including myosin light chain 2 slow (MLC2s). To identify potential cis-acting DNA sites regulating MLC2s following ablation, plasmid constructs were transfected in vivo into regenerating overloaded plantaris muscles. Activity of the 270bp promoter (-270MLC2s) was increased in OV muscles at 28 days. Mutation of the MEF2 site (-270MEF2) knocked out the overload-induced activity of the promoter. Mutation of the Ebox (-270Ebox) resulted in an earlier induction with OV and mutation of the CACC site (-270CACC) resulted in increased activity in the CON PLN with OV induction detected by 21 days. These results demonstrate that the -270MLC2s promoter contains the elements necessary for expression of MLC2s in regenerating OV PLN. More importantly, mutation analysis of -270MLC2s promoter demonstrates that mechanical loading induced expression shares some common molecular mechanisms with slow nerve dependent model regulation. In these two models of physiological induction of MLC2s, the CACC site acts as a repressor region (on/off switch) and the MEF2 site acts to modulate quantitative expression.

Levitt, D. S. (2003). "Apicoectomy of an endosseous implant to relieve paresthesia: a case report." Implant Dent 12(3): 202-5.
 Various radiographic and surgical techniques have been recommended to avoid paresthesia following mandibular implant placement. However, nerve impingement is sometimes inevitable, and when lingering numbness is reported, clinicians have a limited number of corrective options. This report describes a technique for cutting-back the apex of the implant, a technique that may be useful when lingering numbness persists after osseointegration has occurred.

Lewin-Kowalik, J., W. Marcol, et al. (2003). "Dead-ended autologous connective tissue chambers in peripheral nerve repair--early observations." Acta Physiol Hung 90(2): 157-66.
 The effects of the repair of nerve gap injuries are still unsatisfactory, despite the great progress in microsurgery. Until now, there is no effective method to induce the regeneration of the transected peripheral nerve when its distal stump is missing. The aim of this work was to examine whether the implantation of dead-ended connective tissue chambers can promote the outgrowth of injured peripheral neurites. This method differs from all previous nerve guides because it totally eliminates the distal part of the nerve and restricts the influence of surrounding tissues. We have also tried to establish whether some neurotrophic factors can be applied by means of these chambers. The results of this work show that dead-ended autologous connective tissue chambers can be a useful tool in peripheral nerve injuries treatment, even when the distal part of the nerve is missing.

Li, F., D. Carlsson, et al. (2003). "Cellular and nerve regeneration within a biosynthetic extracellular matrix for corneal transplantation." Proc Natl Acad Sci U S A.
 Our objective was to determine whether key properties of extracellular matrix (ECM) macromolecules can be replicated within tissue-engineered biosynthetic matrices to influence cellular properties and behavior. To achieve this, hydrated collagen and N-isopropylacrylamide copolymer-based ECMs were fabricated and tested on a corneal model. The structural and immunological simplicity of the cornea and importance of its extensive innervation for optimal functioning makes it an ideal test model. In addition, corneal failure is a clinically significant problem. Matrices were therefore designed to have the optical clarity and the proper dimensions, curvature, and biomechanical properties for use as corneal tissue replacements in transplantation. In vitro studies demonstrated that grafting of the laminin adhesion pentapeptide motif, YIGSR, to the hydrogels promoted epithelial stratification and neurite in-growth. Implants into pigs' corneas demonstrated successful in vivo regeneration of host corneal epithelium, stroma, and nerves. In particular, functional nerves were observed to rapidly regenerate in implants. By comparison, nerve regeneration in allograft controls was too slow to be observed during the experimental period, consistent with the behavior of human cornea transplants. Other corneal substitutes have been produced and tested, but here we report an implantable matrix that performs as a physiologically functional tissue substitute and not simply as a prosthetic device. These biosynthetic ECM replacements should have applicability to many areas of tissue engineering and regenerative medicine, especially where nerve function is required.

Li, Y., N. Irwin, et al. (2003). "Axon regeneration in goldfish and rat retinal ganglion cells: differential responsiveness to carbohydrates and cAMP." J Neurosci 23(21): 7830-8.
 Mammalian retinal ganglion cells (RGCs) do not normally regenerate their axons through an injured optic nerve, but can be stimulated to do so by activating macrophages intraocularly. In a cell culture model of this phenomenon, we found that a small molecule that is constitutively present in the vitreous, acting in concert with macrophage-derived proteins, stimulates mature rat RGCs to regenerate their axons if intracellular cAMP is elevated. In lower vertebrates, RGCs regenerate their axons spontaneously in vivo, and in culture, the most potent axon-promoting factor for these cells is a molecule that resembles the small vitreous-derived growth factor from the rat. This molecule was isolated chromatographically and was shown by mass spectrometry to be a carbohydrate. In agreement with this finding, D-mannose proved to be a potent axon-promoting factor for rat RGCs (ED50 approximately 10 microm); this response was cAMP-dependent and was augmented further by macrophage-derived proteins. Goldfish RGCs showed far less selectivity, responding strongly to either D-mannose or D-glucose in a cAMP-independent manner. These findings accord well with the success or failure of optic nerves to regenerate in higher and lower vertebrates in vivo. The axon-promoting effects of mannose are highly specific and are unrelated to energy metabolism or glycoprotein synthesis.

Li, L., Q. Xu, et al. (2003). "Combined therapy of methylprednisolone and brain-derived neurotrophic factor promotes axonal regeneration and functional recovery after spinal cord injury in rats." Chin Med J (Engl) 116(3): 414-8.
 OBJECTIVE: To investigate the effects of combination therapy with methylprednisolone (MP) and brain-derived neurotrophic factor (BDNF) on axonal remyelination and functional recovery after spinal cord injury in rats. METHODS: Forty-five rats were randomly divided into three groups: Group A received MP and BDNF; group B received MP and cerebrospinal fluid (CSF); and group C received CSF only. Contusion injury to adult rat spinal cord was produced at the T(10) vertebra level followed by immediate intravenous MP or CSF, and was thereafter infused intrathecally with BDNF or CSF for 6 weeks. Axonal remyelination and functional recovery was observed using RT-PCR, immunohistochemistry and open field locomotion. RESULTS: An increase of 28.4% +/- 2.3% in the expression of proteolipid protein (PLP) gene, an endogenous indicator of axonal remyelination, was demonstrated in group A 24 hours after injury. Ten weeks later, there were significant decreases in hematogenous inflammatory cellular infiltration in groups A and B compared to C (P < 0.05). Concomitantly, a significant amount of axonal remyelination was observed in group A compared to groups B and C (P < 0.05). Furthermore, combination therapy using MP and BDNF in group A resulted in stimulation of hindlimb activity as well as improvement in the rate of functional recovery in open field locomotion (P < 0.05). CONCLUSIONS: Combined therapy of MP and BDNF can improve functional recovery through mechanisms that include attenuating inflammatory cellular infiltration and enhancing axonal remyelination at the injury site. Such a combination may be an effective approach for treatment of spinal cord injury.

Li, Y. and C. Owyang (2003). "Musings on the wanderer: what's new in our understanding of vago-vagal reflexes? V. Remodeling of vagus and enteric neural circuitry after vagal injury." Am J Physiol Gastrointest Liver Physiol 285(3): G461-9.
 The vago-vagal reflexes mediate a wide range of digestive functions such as motility, secretion, and feeding behavior. Previous articles in this series have discussed the organization and functions of this important neural pathway. The focus of this review will be on some of the events responsible for the adaptive changes of the vagus and the enteric neutral circuitry that occur after vagal injury. The extraordinary plasticity of the neural systems to regain functions when challenged with neural injury will be discussed. In general, neuropeptides and transmitter-related enzymes in the vagal sensory neurons are downregulated after vagal injury to protect against further injury. Conversely, molecules previously absent or present at low levels begin to appear or are upregulated and are available to participate in the survival-regeneration process. Neurotrophins and other related proteins made at the site of the lesion and then retrogradely transported to the soma may play an important role in the regulation of neuropeptide phenotype expression and axonal growth. Vagal injury also triggers adaptive changes within the enteric nervous system to minimize the loss of gastrointestinal functions resulting from the interruption of the vago-vagal pathways. These may include rearrangement of the enteric neural circuitry, changes in the electrophysiological properties of sensory receptors in the intramural neural networks, an increase in receptor numbers, and changes in the affinity states of receptors on enteric neurons.

Li, Y., Y. Sauve, et al. (2003). "Transplanted olfactory ensheathing cells promote regeneration of cut adult rat optic nerve axons." J Neurosci 23(21): 7783-8.
 Transplantation of olfactory ensheathing cells into spinal cord lesions promotes regeneration of cut axons into terminal fields and functional recovery. This repair involves the formation of a peripheral nerve-like bridge in which perineurial-like fibroblasts are organized into a longitudinal stack of parallel tubular channels, some of which contain regenerating axons enwrapped by Schwann-like olfactory ensheathing cells. The present study examines whether cut retinal ganglion cell axons will also respond to these cells, and if so, whether they form the same type of arrangement. In adult rats, the optic nerve was completely severed behind the optic disc, and a matrix containing cultured olfactory ensheathing cells was inserted between the proximal and distal stumps. After 6 months, the transplanted cells had migrated for up to 10 mm into the distal stump. Anterograde labeling with cholera toxin B showed that cut retinal ganglion cell axons had regenerated through the transplants, entered the distal stump, and elongated for 10 mm together with the transplanted cells. Electron microscopy showed that a peripheral nerve-like tissue had been formed, similar to that seen in the spinal cord transplants. However, in contrast to the spinal cord, the axons did not reach the terminal fields, but terminated in large vesicle-filled expansions beyond which the distal optic nerve stump was reduced to a densely interwoven mass of astrocytic processes.

Li, S., Q. Pan, et al. (2003). "Neurotization of oculomotor, trochlear and abducent nerves in skull base surgery." Chin Med J (Engl) 116(3): 410-3.
 OBJECTIVE: To anatomically reconstruct the oculomotor nerve, trochlear nerve, and abducent nerve by skull base surgery. METHODS: Seventeen cranial nerves (three oculomotor nerves, eight trochlear nerves and six abducent nerves) were injured and anatomically reconstructed in thirteen skull base operations during a period from 1994 to 2000. Repair techniques included end-to-end neurosuture or fibrin glue adhesion, graft neurosuture or fibrin glue adhesion. The relationships between repair techniques and functional recovery and the related factors were analyzed. RESULTS: Functional recovery began from 3 to 8 months after surgery. During a follow-up period of 4 months to 6 years, complete recovery of function was observed in 6 trochlear nerves (75%) and 4 abducent nerves (67%), while partial functional recovery was observed in the other cranial nerves including 2 trochlear nerves, 2 abducent nerves, and 3 oculomotor nerves. CONCLUSIONS: Complete or partial functional recovery could be expected after anatomical neurotization of an injured oculomotor, trochlear or abducent nerve. Our study demonstrated that, in terms of functional recovery, trochlear and abducent nerves are more responsive than oculomotor nerves, and that end-to-end reconstruction is more efficient than graft reconstruction. These results encourage us to perform reconstruction for a separated cranial nerve as often as possible during skull base surgery.

Li, S. and S. M. Strittmatter (2003). "Delayed systemic Nogo-66 receptor antagonist promotes recovery from spinal cord injury." J Neurosci 23(10): 4219-27.
 Traumatized axons possess an extremely limited ability to regenerate within the adult mammalian CNS. The myelin-derived axon outgrowth inhibitors Nogo, oligodendrocyte-myelin glycoprotein, and myelin-associated glycoprotein, all bind to an axonal Nogo-66 receptor (NgR) and at least partially account for this lack of CNS repair. Although the intrathecal application of an NgR competitive antagonist at the time of spinal cord hemisection induces significant regeneration of corticospinal axons, such immediate local therapy may not be as clinically feasible for cases of spinal cord injury. Here, we consider whether this approach can be adapted to systemic therapy in a postinjury therapeutic time window. Subcutaneous treatment with the NgR antagonist peptide NEP1-40 (Nogo extracellular peptide, residues 1-40) results in extensive growth of corticospinal axons, sprouting of serotonergic fibers, upregulation of axonal growth protein SPRR1A (small proline-rich repeat protein 1A), and synapse re-formation. Locomotor recovery after thoracic spinal cord injury is enhanced. Furthermore, delaying the initiation of systemic NEP1-40 administration for up to 1 week after cord lesions does not limit the degree of axon sprouting and functional recovery. This indicates that the regenerative capacity of transected corticospinal tract axons persists for weeks after injury. Systemic Nogo-66 receptor antagonists have therapeutic potential for subacute CNS axonal injuries such as spinal cord trauma.

Libersa, P., D. Roze, et al. (2003). "Preliminary results and evidence of early regeneration in inferior alveolar nerve fibers." Surg Radiol Anat 24(6): 354-7.
 Inferior alveolar nerve (IAN) damage can occur in trauma, cyst enucleation, sagittal split osteotomy or third molar removal, and the consequences are a loss of sensation to the mandibular teeth, gingiva and lower lip. Because of its anatomical position in a bony canal, IAN suture is rarely evoked. The aim of this study was to demonstrate the reality of IAN regeneration by using electrophysiological and histological methods after experimental section and suture of this nerve in rabbits. Nine adult female animals were used for the experiments. Six months after section and suturing using 10.0 nylon with a conventional technique, electrical stimulation of the nerve was performed to record electrophysiological activity. Each rabbit was its own reference. In each case, an action potential was recorded after microsurgical repair and definitively suppressed by section of the nerve. Morphometric analysis showed a decrease in the number of nerve fibers in the operated nerve versus the control nerve. The histological study showed an increase in nerve fibers with a cross-sectional area of 19-36 and 37-73 micro m(2) and a decrease in the smaller fibers (2-4 and 5-7 micro m(2)). This preliminary study confirms the possibility of nerve regeneration in rabbits 6 months after section and conventional suturing.

Liebkind, R., T. Laatikainen, et al. (2003). "Is the soluble KDI domain of gamma1 laminin a regeneration factor for the mammalian central nervous system?" J Neurosci Res 73(5): 637-43.
 Regeneration of adult mammalian CNS is poor as a result of environmental factors that prevent axon growth. The major factors hampering regeneration of central axons include proteins released from the damaged myelin sheets of the injured neuronal pathways and formation of the glial scar. By using an experimental model of human CNS injury, we show that survival and neurite outgrowth of human central neurons are significantly enhanced by the soluble KDI domain of gamma1 laminin. Our results indicate that the KDI domain appears to neutralize both glia-derived inhibitory signals and inhibitory molecules released from the myelin of the adult human spinal cord. We propose that the KDI domain may enhance regeneration of injuries in the adult mammalian CNS.

Lin, G., K. C. Chen, et al. (2003). "Neurotrophic effects of vascular endothelial growth factor and neurotrophins on cultured major pelvic ganglia." BJU Int 92(6): 631-5.
 OBJECTIVE: To investigate the feasibility of using a ganglial culture system to screen various growth factors as potential therapeutic agents for pelvic nerve injuries. MATERIALS AND METHODS: The major pelvic ganglia (MPG) were isolated from male rats and attached to culture dishes with the aid of Matrigel (Becton Dickinson, Mountain View, CA, USA). Alternatively, the dorso-caudal region (DCR) of MPG, from which the cavernous nerves originate, was dissected and then attached to a Matrigel-coated coverslip. The MPG or DCR was cultured in serum-free medium supplemented with phosphate-buffered saline (PBS, control), 50 ng/mL of vascular endothelial growth factor (VEGF), 20 ng/mL of a neurotrophin (BDNF, NT3, or NT4), or combinations of these growth factors. After 2 days of incubation, the ganglial tissues with their outgrowing nerve fibres were stained for the expression of NADPH-diaphorase, tyrosine hydroxylase (TH) and acetylcholinesterase (AChE). The length and staining intensity of nerve fibres were analysed. RESULTS: The outgrowing fibres were significantly longer in MPG treated with any of the four tested growth factors than in PBS-treated MPG. The combination of VEGF and NT3 induced the best fibre growth. Improvements to the culturing conditions allowed a histological examination of the outgrowing fibres for the expression of nitric oxide synthase (NOS), TH and AChE. VEGF and BDNF were equally capable of inducing NOS- and TH-expressing fibres. BDNF was much weaker than VEGF for inducing AChE-expressing fibres. CONCLUSIONS: This improved culturing system is potentially useful for screening nerve-regenerating factors; VEGF had neurotrophic effects comparable with BDNF, NT3, or NT4.

Lin, H., J. Bao, et al. (2003). "Rapid electrical and delayed molecular signals regulate the serum response element after nerve injury: convergence of injury and learning signals." J Neurobiol 57(2): 204-20.
 Axotomy elicits changes in gene expression, but little is known about how information from the site of injury is communicated to the cell nucleus. We crushed nerves in Aplysia californica and the sciatic nerve in the mouse and found short- and long-term activation of an Elk1-SRF transcription complex that binds to the serum response element (SRE). The enhanced short-term binding appeared rapidly and was attributed to the injury-induced activation of an Elk1 kinase that phosphorylates Elk1 at ser383. This kinase is the previously described Aplysia (ap) ERK2 homologue, apMAPK. Nerve crush evoked action potentials that propagated along the axon to the cell soma. Exposing axons to medium containing high K(+), which evoked a similar burst of spikes, or bathing the ganglia in 20 microM serotonin (5HT) for 20 min, activated the apMAPK and enhanced SRE binding. Since 5HT is released in response to electrical activity, our data indicate that the short-term process is initiated by an injury-induced electrical discharge that causes the release of 5HT which activates apMAPK. 5HT is also released in response to noxious stimuli for aversive learning. Hence, apMAPK is a point of convergence for injury signals and learning signals. The delay before the onset of the long-term SRE binding was reduced when the crush was closer to the ganglion and was attributed to an Elk1 kinase that is activated by injury in the axon and retrogradely transported to the cell body. Although this Elk1 kinase phosphorylates mammalian rElk1 at ser383, it is distinct from apMAPK.

Lipson, A. C., J. Widenfalk, et al. (2003). "Neurotrophic properties of olfactory ensheathing glia." Exp Neurol 180(2): 167-71.
 Olfactory ensheathing cells (OEC) constitute a specialized population of glia that accompany primary olfactory axons and have been reported to facilitate axonal regeneration after spinal cord injury in vivo. In the present report we describe OEC neurotrophic factor expression and neurotrophic properties of OECs in vitro. Investigation of the rat olfactory system during development and adulthood by radioactive in situ hybridization revealed positive labeling in the olfactory nerve layer for the neurotrophic molecules S-100beta, CNTF, BMP-7/OP-1, and artemin, as well as for the neurotrophic factor receptors RET and TrkC. Ribonuclease protection assay of cultured OEC revealed expression of NGF, BDNF, GDNF, and CNTF mRNA, while NT3 and NT4 mRNA were not detectable. In vitro bioassays of neurotrophic activity involved coculturing of adult OEC with embryonic chick ganglia and demonstrated increased neurite outgrowth from sympathetic, ciliary, and Remak's ganglia. However, when culturing the ganglia with OEC-conditioned medium, neurite outgrowth was not stimulated to any detectable extent. Our results suggest that the neurotrophic properties of OEC may involve secretion of neurotrophic molecules but that cellular interactions are crucial.

Liu, H., M. Dong, et al. (2003). "[An study on functioning remobilization of the paralyzed vocal cord by latero-terminal neurorrhaphy in rats]." Lin Chuang Er Bi Yan Hou Ke Za Zhi 17(9): 554-6.
 OBJECTIVE: To investigate the effect and the clinic value of latero-terminal neurorrhaphy to treat vocal cord paralysis. METHOD: Sixty SD rats were divided into three groups. In the experimental group, the right recurrent laryngeal nerve(RLN) was incised and anastomosed to the right phren nerve by means of latero-terminal neurorrhaphy. The internal nerve of the right RLN was incised and anastomosed to the right ansa cervicals nerve by end-to-end nerve anastomosis. In control group, the right RLN was incised and sutured to the right phren nerve by end-to-end nerve anastomosis. The internal nerve of the right RLN were incised and anastomosed to the right ansa cervicals nerve by end-to-end nerve anastomosis. In normal group rats, the nerves were only exposed. One to three months later, 10 rats from each group were examined for vocal cord movement and nerve regeneration by using fibrolaryngscope and nerve electromyography. RESULT: One months after operation. This effect of latero-terminal neurorrhaphy had significant difference compared with the control group (P < 0.05). Three months after operation. This effect of latero-terminal neurorrhaphy had not significant difference compared with the control group (P > 0.05). CONCLUSION: The latero-terminal neurorrhaphy has a similar treatment effect compared with end-to-end nerve anastomosis. This microsurgical technique provides a new method for treating vocal cord paralysis.

Liu, M. Y., J. P. Chiang, et al. (2003). "Abamectin attenuates gastric mucosal damage induced by ethanol through activation of vagus nerve in rats." Alcohol 30(1): 61-5.
 Some type A gamma-aminobutyric acid (GABA(A)) receptor agonists are effective in protecting against the formation of stomach lesions induced by ethanol. Natural product abamectin, one of the existing GABA(A) receptor agonists, might protect against the development of gastric ulcers induced by ethanol. We investigated the protective effect of abamectin against the formation of gastric mucosal lesions induced by ethanol in rats. Abamectin (3 mg/kg, p.o.) was given to rats 1 h before administration of ethanol [4 ml of a 30% (volume/volume) solution]. Mucosal lipid peroxidation (LPO), nitric oxide (NO) levels, and ulcer index were measured 3 h after gastric surgery (vagotomy vs. sham vagotomy) in treated versus control subjects. Abamectin attenuated ethanol-induced gastric ulceration, decreased LPO regeneration, and increased NO production in the gastric mucosa of rats in the sham vagotomy group. However, this protective effect of abamectin against ethanol-induced gastric lesions was not observed in rats in the group that underwent vagotomy. These results support the suggestion that administration of abamectin ameliorated the ethanol-induced gastric mucosal injury through elevation of NO production. Activation of the vagus nerve may be involved in the abamectin-associated gastric protection against the effects of ethanol in rats.

Liu, S., P. Damhieu, et al. (2003). "Efficient reinnervation of hindlimb muscles by thoracic motor neurons after nerve cross-anastomosis in rats." J Neurosurg 99(5): 879-85.
 OBJECT: Peripheral motor axons can regenerate through motor endoneurial tubes of foreign nerves to reinnervate different target muscles. This regenerative capacity has been brought to clinical applications for restorative surgery after nerve or root injury. In this study the authors explore the extent to which nerve cross-anastomosis between lower intercostal nerves and lumbar ventral roots would be effective in inducing reinnervation of paralyzed hindlimb muscles after spinal cord hemisection at the thoracolumbar boundary in rats. METHODS: The proximal extremities of sectioned intercostal nerves T10-12 were surgically connected to the distal extremities of sectioned ipsilateral lumbar ventral roots L3-5, respectively. Motor activity reappeared 2 months postsurgery; however, locomotion was not restored and inappropriate motor patterns persisted at 9 months postsurgery. At that time, data from electrophysiological and histological studies and horseradish peroxidase retrograde labeling demonstrated efficient regrowth of thoracic motor neuron axons that reached hindlimb muscles. They also revealed a persistent maturation defect of regrown fibers, as shown by size heterogeneity and presumable extensive axonal branching. These features are consistent with reduced neural activity subsequent to continuing inappropriate motor patterns. CONCLUSIONS: These results indicate that cross-anastomosis of intercostal nerves with lumbar ventral roots allows efficient reinnervation of paralyzed hindlimb muscles after spinal cord hemisection in rats. Stimulating the reorganization of the neuronal circuitry in the central nervous system by locomotion training or other methods would presumably result in both functional and anatomical improvements. This experimental setting provides a convenient animal model to investigate these processes.

Liu, Y., Y. Ishida, et al. (2003). "Effects of repeated stress on regeneration of serotonergic and noradrenergic axons in the cerebral cortex of adult rats." Neurosci Lett 339(3): 227-30.
 We examined whether regeneration of serotonergic (5-HT) and noradrenergic (NA) axons might be affected by stress. Neurotoxins to 5-HT or NA axons were injected into the frontal cortex to cause partial denervation. Mild restraint stress (40 min/day) was started 16 days later and given for 14 consecutive days (30-day stressed group). Non-stressed animals were divided into two groups, animals sacrificed at 14 days (14-day control group) and those sacrificed at 30 days (30-day control group) after the toxin injection. In immunohistochemical study, the denervation area of 5-HT axons but not that of NA axons was significantly smaller in the 30-day control group than in the 14-day control group. However, there was no significant difference between the 14-day control and 30-day stressed groups. These findings suggested that regeneration of 5-HT axons occurred earlier than that of NA axons, and that stress exerted inhibitory influence on regeneration of 5-HT axons.

Loiacono, C. M., N. S. Taus, et al. (2003). "The herpes simplex virus type 1 ICP0 promoter is activated by viral reactivation stimuli in trigeminal ganglia neurons of transgenic mice." J Neurovirol 9(3): 336-45.
 Herpes simplex virus type 1 (HSV-1) causes a latent infection in sensory ganglia neurons in humans and in the mouse model. The ability of the virus to latently infect neurons and reactivate is central to the ability of HSV-1 to remain in the human population and spread to new hosts. It is possible that neuronal transcriptional proteins control latency and reactivation by modulating activation of the HSV-1 immediate-early (IE) gene ICP0. We have previously shown that factors in trigeminal ganglia neurons can differentially activate the IE ICP0 promoter and the IE ICP4 promoter in developing trigeminal ganglia neurons of transgenic mice. Ultraviolet (UV) irradiation and hyperthermic stress have been shown to result in HSV-1 reactivation from sensory neurons in the mouse model. Reporter transgenic mice were exposed to UV irradiation or hyperthermia to test whether stimuli that are known to reactivate HSV-1 could activate viral IE promoters in the absence of viral proteins. Measurement of beta-galactosidase activity in trigeminal ganglia from these transgenic mice indicated that the ICP0 promoter activity was significantly increased by both UV irradiation and hyperthermia. The IE genes ICP4 and ICP27 and the late gene gC reporter transgenes failed to be activated in parallel experiments. These results suggest that the ICP0 promoter is a target for activation by host transcription factors in sensory neurons that have undergone damage. It further suggests the possibility that activation of ICP0 gene expression by neuronal transcription factors may be important in reactivation of HSV-1 in neurons.

Lopez, I., C. Ayala, et al. (2003). "Synaptophysin immunohistochemistry during vestibular hair cell recovery after gentamicin treatment." Audiol Neurootol 8(2): 80-90.
 In the present study, morphometric and immunohistochemical techniques were used to evaluate the degree of synaptic recovery in the chinchilla crista sensory epithelia during various post-gentamicin-treatment periods of hair cell loss and recovery. For this purpose, two groups of animals were treated with Gelfoam pellets impregnated with 50 micro g of gentamicin implanted in the perilymphatic space within the otic capsule of the superior semicircular canal. Animals were sacrificed 1, 2 and 4 weeks after treatment. The degree of synaptic reinnervation was evaluated in the horizontal crista of the first group of animals using immunohistochemical techniques and antibodies against synaptophysin, a marker for synaptic reinnervation and synaptogenesis. Quantification of immunoreactivity in this group was made in the mid-region of the crista using the NIH 'Image' program. The second group of animals was used for quantification of the number of hair cells and supporting cells in the horizontal crista. In the normal sensory epithelium, synaptophysin immunoreactivity was found in the areas corresponding to the known distribution of afferent and efferent nerve terminals. Immunoreactivity was predominantly located within the afferent calyces of type I hair cells. No immunoreactivity was found in the supporting cells. Seven days after treatment there was a significant loss of hair cells and synaptophysin-stained area (SSA). In the mid-region of the crista the loss of synaptophysin immunoreactivity was quantitatively the greatest within the central zone of this region (93%) while the loss of hair cells was the smallest. These results suggest that afferent and efferent nerve terminals were also severely affected by the ototoxic treatment. Four weeks after treatment corresponding to the end of the recovery phase of gentamicin ototoxicity, there was a proportional increase in the number of hair cells and of the degree of SSA in the mid-region of the crista. The number of hair cells recovered to 58% with a recovery of SSA to 54% of normal. These results suggest that a greater fraction of synaptophysin expression within the sensory epithelium depends on the presence of afferent calyceal endings, which are greatly affected by gentamicin. Also, these results demonstrate a significant level of reinnervation of the newly regenerated hair cells, forecasting the potential for functionality of the regenerated hair cells.

Love, F. M., Y. J. Son, et al. (2003). "Activity alters muscle reinnervation and terminal sprouting by reducing the number of Schwann cell pathways that grow to link synaptic sites." J Neurobiol 54(4): 566-76.
 In partially denervated rodent muscle, terminal Schwann cells (TSCs) located at denervated end plates grow processes, some of which contact neighboring innervated end plates. Those processes that contact neighboring synapses (termed "bridges") appear to initiate nerve terminal sprouting and to guide the growth of the sprouts so that they reach and reinnervate denervated end plates. Studies conducted prior to knowledge of this potential involvement of Schwann cells showed that direct muscle stimulation inhibits terminal sprouting following partial denervation (Brown and Holland, 1979). We have investigated the possibility this inhibition results from an alteration in the growth of TSC processes. We find that stimulation of partially denervated rat soleus muscle does not alter the length or number of TSC processes but does reduce the number of TSC bridges. Stimulation also reduces the number of TSC bridges that form between end plates during reinnervation of a completely denervated muscle. The nerve processes ("escaped fibers") that normally grow onto TSC processes during reinnervation are also reduced in length. Therefore, stimulation alters at least two responses to denervation in muscles: (1) the ability of TSC processes to form or maintain bridges with innervated synaptic sites, and (2) the growth of axons along processes extended by TSCs.

Low, H. L., A. Nogradi, et al. (2003). "Axotomized motoneurons can be rescued from cell death by peripheral nerve grafts: the effect of donor age." J Neuropathol Exp Neurol 62(1): 75-87.
 Injury to neonatal nerves, unlike adult nerves, results in poor regeneration and extensive motoneuron death. We examined whether exposure to a more mature nerve environment could rescue axotomized motoneurons following neonatal injury. The sciatic nerve in 1 hindlimb of 3-day-old (P3) rats was transected and the cut end sutured to a nerve graft taken from donor rats, which ranged between P3 and P21. The extent of motoneuron survival and axon regeneration was established 7 days later. Since integrins play an important role in regeneration, we also examined the effect of manipulating integrin binding in nerve grafts. Following axotomy at P3 and implantation of nerve grafts from 3-day-old rats, approximately 38% of motoneurons survived. In contrast, grafts from rats aged 5 days and older resulted in an improvement in regeneration, and over 70% of motoneurons survived. This survival-promoting effect of P5 grafts was prevented by blocking beta1-integrins. In contrast, increasing beta1-integrin levels in grafts from P3 rats dramatically increased motoneuron survival. Thus, following neonatal nerve injury, exposure to a more mature nerve environment significantly increases motoneuron survival, an effect that is dependent upon beta1-integrin signaling. Therefore, pharmacological upregulation of beta1-integrins may significantly improve the outcome of neonatal nerve injuries.

Lu, Q., Q. Cui, et al. (2003). "c-Jun expression in surviving and regenerating retinal ganglion cells: effects of intravitreal neurotrophic supply." Invest Ophthalmol Vis Sci 44(12): 5342-8.
 PURPOSE: To investigate c-jun expression in surviving and axon-regenerating retinal ganglion cells (RGCs) and the effect of intravitreal neurotrophic supply on c-jun expression. METHODS: All animals underwent optic nerve transection (ONT) 0.5 mm behind the eyeball. Some animals underwent a replacement of the optic nerve with an autologous sciatic nerve graft (SNG) to allow axonal regrowth. To provide a neurotrophic supply, a peripheral nerve (PN) segment or brain-derived neurotrophic factor (BDNF)/ciliary neurotrophic factor (CNTF) was applied intravitreally. The time course of c-jun expression was first examined in both surviving and regenerating RGCs. Then, c-jun expression was examined in surviving and regenerating RGCs 3 weeks after intravitreal BDNF/CNTF treatment. Animals with vehicle eye injection were used as the control. Fluorescent dye was used for retrograde labeling of surviving (applied behind the eyeball) and regenerating (applied at the distal end of the SNG) RGCs. All retinas were immunohistochemically stained for c-jun. RESULTS: c-Jun was not detected in normal RGCs, but weak expression was seen in surviving RGCs after ON injury. The proportion of c-jun-positive (+) RGCs among surviving cell population was 52.6% to 86.5% 2 to 6 weeks after ONT. Among regenerating RGCs, more than 80% expressed c-jun in all treatment groups, a proportion that was significantly higher after CNTF treatment (90.7%). In addition, c-jun expression was much stronger in intensity and the c-jun(+) nuclei were much larger in regenerating than in surviving RGCs. CONCLUSIONS: c-Jun expression in RGCs was upregulated after injury. Most regenerating RGCs were c-jun(+), and the intensity of c-jun expression was higher in regenerating than in surviving RGCs. CNTF also upregulated c-jun expression in RGCs.

Lu, P., L. L. Jones, et al. (2003). "Neural stem cells constitutively secrete neurotrophic factors and promote extensive host axonal growth after spinal cord injury." Exp Neurol 181(2): 115-29.
 Neural stem cells (NSCs) offer the potential to replace lost tissue after nervous system injury. This study investigated whether grafts of NSCs (mouse clone C17.2) could also specifically support host axonal regeneration after spinal cord injury and sought to identify mechanisms underlying such growth. In vitro, prior to grafting, C17.2 NSCs were found for the first time to naturally constitutively secrete significant quantities of several neurotrophic factors by specific ELISA, including nerve growth factor, brain-derived neurotrophic factor, and glial cell line-derived neurotrophic factor. When grafted to cystic dorsal column lesions in the cervical spinal cord of adult rats, C17.2 NSCs supported extensive growth of host axons of known sensitivity to these growth factors when examined 2 weeks later. Quantitative real-time RT-PCR confirmed that grafted stem cells expressed neurotrophic factor genes in vivo. In addition, NSCs were genetically modified to produce neurotrophin-3, which significantly expanded NSC effects on host axons. Notably, overexpression of one growth factor had a reciprocal effect on expression of another factor. Thus, stem cells can promote host neural repair in part by secreting growth factors, and their regeneration-promoting activities can be modified by gene delivery.

Luk, H. W., L. J. Noble, et al. (2003). "Macrophages contribute to the maintenance of stable regenerating neurites following peripheral nerve injury." J Neurosci Res 73(5): 644-58.
 Normal adult uninjured nerve is unable to support axonal regeneration. We have studied the mechanisms underlying the regeneration of peripheral nerve by culturing adult mouse dorsal root ganglia (DRG) explants on unfixed, longitudinal cryosections of either the uninjured sciatic nerve or the distal segment of the transected sciatic nerve. We found that, initially, DRG grew vigorously on cryosections of both uninjured and postinjury sciatic nerves. However, the neurites began to degenerate shortly after contact with the uninjured nerve, whereas those growing on postinjury nerve substrate remained healthy for up to 9 days in culture. This ability to support stable outgrowth peaked at 8 days, gradually decreased by 10 days, and disappeared by 20 days after injury. Macrophages appeared in the distal segment by 4 days postinjury and had infiltrated its entire length by 8 days. Uninjured nerve cryosections could be rendered supportive of stable outgrowth by preincubation with macrophage-conditioned medium or by brief trypsinization. The activity of the macrophage-conditioned medium was augmented upon activation of macrophages. Together these findings suggest that the environment of the sciatic nerve undergoes a transformation during Wallerian degeneration such that it becomes transiently supportive of the stable outgrowth of neurites. This transformation may be mediated by a proteolytic activity, generated by activated macrophages, that removes a putative "degeneration signal" protein normally present in the adult nerve and thus contributes to the maintenance of stable regenerating neurites.

Lukowiak, K., Z. Haque, et al. (2003). "Long-term memory survives nerve injury and the subsequent regeneration process." Learn Mem 10(1): 44-54.
 A three-neuron network (a central pattern generator [CPG]) is both sufficient and necessary to generate aerial respiratory behavior in the pond snail, Lymnaea stagnalis. Aerial respiratory behavior is abolished following a specific nerve crush that results in axotomy to one of the three CPG neurons, RPeD1. Functional regeneration of the crushed neurite occurs within 10 days, allowing aerial respiratory behavior to be restored. Functional regeneration does not occur if the connective is cut rather than crushed. In unaxotomized snails, aerial respiratory behavior can be operantly conditioned, and following memory consolidation, long-term memory (LTM) persists for at least 2 weeks. We used the Lymnaea model system to determine (1) If in naive animals axotomy and the subsequent regeneration result in a nervous system that is competent to mediate associative learning and LTM, and (2) if LTM survives RPeD1 axotomy and the subsequent regenerative process. We show here that (1) A regenerated nervous system is competent to mediate associative memory and LTM, and (2) LTM survives axotomy and the subsequent regenerative process.

Ma, Y., X. Xu, et al. (2003). "[Experimental study on using mGTR and venous autografts to repair facial nerve defects in rabbits]." Shanghai Kou Qiang Yi Xue 12(1): 38-40.
 OBJECTIVE: To observe and compare the effects of using mGTR and venous autografts to repair the defect of facial nerve in rabbits. METHODS: mGTR and venous autografts were performed on three groups of rabbits. Then physical and histological examinations were carried out to get the results of these two repair methods. RESULTS: Both the two methods can induce regeneration of facial nerve. The mGTR is more effective and faster in repairing the nerve defect. Three months after operation, the distance of NCV between bilateral facial nerve in mGTR group was (2.10 +/- 1.2) m/s, (6.80 +/- 1.4) m/s in venous autografts, (2.16 +/- 1.6) m/s in nerve transplantation group. Histologically, the facial nerve fibers were arranged regularly and in continuity in mGTR, the facial nerve fibers were arranged irregularly with marked hyperplasia of connective tissues in venous autografts three months after repair. CONCLUSIONS: The mGTR is more effective than venous autografts in repair of facial nerve.

Ma, Y. P. and Z. H. Pan (2003). "Spontaneous regenerative activity in mammalian retinal bipolar cells: roles of multiple subtypes of voltage-dependent Ca2+ channels." Vis Neurosci 20(2): 131-9.
 Patch-clamp recordings were used to investigate the properties of the regenerative activity in acutely isolated bipolar cells from the rat retina. Spontaneous, pacemaker-like membrane potential oscillations were observed in all rod bipolar cells and the majority of cone bipolar cells. The waveform of the regenerative potential was stereotypical but distinct among different bipolar cell groups, especially between rod and cone bipolar cells. The spontaneous activity was completely blocked by Co2+, suggesting that Ca2+ influx through voltage-dependent Ca2+ channels was required for initiating such activity. Ca2+-induced Ca2+ release, however, was not found to be involved. The spontaneous activity was also blocked by mibefradil, a T-type Ca2+ channel antagonist. In contrast, application of nimodipine, an L-type Ca2+ current antagonist, affected mainly the waveform of the regenerative potential. This study shows that mammalian retinal bipolar cells in isolation are also capable of generating Ca2+-dependent spontaneous regenerative potential. However, T-type Ca2+ channels appear to be essential for the initiation of the spontaneous activity in mammalian bipolar cells.

Ma, J., L. N. Novikov, et al. (2003). "Early nerve repair after injury to the postganglionic plexus: an experimental study of sensory and motor neuronal survival in adult rats." Scand J Plast Reconstr Surg Hand Surg 37(1): 1-9.
 The optimal time for brachial plexus nerve repair is debatable. In this study we examined whether early re-establishment of neurotrophic support from the periphery might reduce neuronal loss. In 14 adult rats, the C7 spinal nerve was transsected. All sensory cells of the dorsal root ganglion and spinal motor neurons projecting into the C7 nerve were labelled retrogradely. The proximal and distal portions of the C7 nerve were then reanastomosed by either primary repair or by a vascularised or conventional ulnar nerve graft. At 16 weeks postoperatively, the nerve repair had significantly reduced the loss of both sensory and motor C7 neurons. Most striking was that a 30% motor neuronal loss in the control was almost eliminated by early nerve repair. In the grafted animals, half of the surviving neurons had regenerated through the graft, with no difference between vascularised and conventional nerve grafts. These results suggest that early surgical intervention may promote neuronal survival and regeneration after injuries to the brachial plexus.

Ma, L., R. J. Carter, et al. (2003). "RAGE is expressed in pyramidal cells of the hippocampus following moderate hypoxic-ischemic brain injury in rats." Brain Res 966(2): 167-74.
 The receptor for advanced glycation end products (RAGE) is a multi-ligand member of the immunoglobulin superfamily of cell surface molecules. The RAGE-ligand interaction has a putative role in a range of chronic disorders and is also known to contribute to both inflammatory/degenerative processes as well as regeneration in peripheral nerve injury. We have investigated the expression of RAGE in the moderate hypoxic-ischemic (HI) rat brain injury model in order to determine if this receptor is involved in the cellular perturbation mediated by ischemic stress. RAGE mRNA levels were detected by in situ hybridization using a DIG-labelled 48 mer oligonucleotide probe. Results showed a high level of expression of RAGE mRNA in the CA1/2 pyramidal cell layer of the hippocampus on the lesioned side of the brain 72 h after a moderate hypoxic-ischemic insult. RAGE was not expressed on the control side of the hippocampus. The RAGE-positive cells had a unique morphology, being angular in shape and atrophied with a condensed cell nucleus. They were NeuN-positive and were identified as dying cells by staining with thionin/acid fuchsin. A subset of cells was positive for cleaved Caspase-3, a marker for apoptosis. Together these data show that RAGE is expressed in dying neurons and suggest that RAGE may have a role in neuronal cell death mediated by ischemic stress. Identification of the ligand for RAGE in the ischemic brain may lead to a better understanding of RAGE-mediated cellular dysfunction in the CNS.

MacDonald, S. C., I. G. Fleetwood, et al. (2003). "Functional motor neurons differentiating from mouse multipotent spinal cord precursor cells in culture and after transplantation into transected sciatic nerve." J Neurosurg 98(5): 1094-103.
 OBJECT: One of the current challenges in neurobiology is to ensure that neural precursor cells differentiate into specific neuron types, so that they can be used for transplantation purposes in patients with neuron loss. The goal of this study was to determine if spinal cord precursor cells could differentiate into motor neurons both in culture and following transplantation into a transected sciatic nerve. METHODS: In cultures with trophic factors, neurons differentiate from embryonic precursor cells and express motor neuronal markers such as choline acetyltransferase (ChAT), Islet-1, and REG2. Reverse transcription-polymerase chain reaction analysis has also demonstrated the expression of Islet-1 in differentiated cultures. A coculture preparation of neurospheres and skeletal myocytes was used to show the formation of neuromuscular connections between precursor cell-derived neurons and myocytes both immunohistochemically and electrophysiologically. Following various survival intervals, precursor cells transplanted distal to a transection of the sciatic nerve differentiated into neurons expressing the motor neuron markers ChAT and the alpha1 1.2 (class C, L-type) voltage-sensitive Ca++ channel subunit. These cells extended axons into the muscle, where they formed cholinergic terminals. CONCLUSIONS: These results demonstrate that motor neurons can differentiate from spinal cord neural precursor cells grown in culture as well as following transplantation into a transected peripheral nerve.

Mackiewicz, Z., M. Hukkanen, et al. (2003). "Dual effects of caspase-1, interleukin-1 beta, tumour necrosis factor-alpha and nerve growth factor receptor in inflammatory myopathies." Clin Exp Rheumatol 21(1): 41-8.
 OBJECTIVE: To analyse the expression of factors potentially involved in skeletal muscle degeneration and regeneration in dermatomyositis (DM), systemic sclerosis (SSc), polymyositis (PM), systemic lupus erythematosus (SLE) and non-inflammatory myopathies. METHODS: Immunohistochemical staining of skeletal muscle biopsies (10 DM, 10 SSc, 10 PM, 10 SLE, 10 non-inflammatory myopathies) for tumour necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL-1 beta), activated caspase-1, pan-macrophage marker CD68, inducible nitric oxide synthase (NOS2) and nerve growth factor receptor (NGFR). TechMate staining robot and biotin-streptavidin protocol were used. RESULTS: Expression of TNF-alpha, IL-1 beta, caspase-1 and NOS2 was found in the cytoplasm and sarcolemma of dystrophic skeletal muscle fibres. TNF-alpha and IL-1 beta immunoreactive profiles were faint and few and close to satellite nuclei-containing regenerating muscle fibres both in inflammatory and non-inflammatory myopathies. NGFR expression was found in comparable areas. In non-inflammatory inherited myopathies more nuclei were caspase-1 immunoreactive whereas caspase-1 expression was rarely seen in inflammatory myopathies, implying regeneration of the affected muscle fibres. CONCLUSION: Prominent expression of the proinflammatory factors TNF-alpha, IL-1 beta and NOS2 and caspase-1 is associated with muscle fibre damage, albeit when expressed to a low degree these factors may, like NGFR, contribute to muscle regeneration and healing.

Madaschi, L., A. M. Di Giulio, et al. (2003). "Muscle reinnervation and IGF-I synthesis are affected by exposure to heparin: an effect partially antagonized by anti-growth hormone-releasing hormone." Neurochem Res 28(1): 163-8.
 Sciatic nerve crush was performed in 2-day-old rats, then reinnervation of the extensor digitorum longus muscle, motor neuron survival, and muscle IGF-I production were monitored. In saline-treated rats, the extent of reinnervation was around 50% and the number of EDL reinnervating motor neurons was significantly reduced. In heparin-treated rats the extent of muscle reinnervation, the recovery of nerve-evoked muscle twitch tension, and the number of motor neurons reinnervating the extensor digitorum longus muscle were greatly enhanced compared to saline-treated rats. In addition, treatment with heparin increased markedly insulin-like growth factor-I levels in denervated muscles. The concomitant exposure to anti-growth hormone releasing hormone partially abolished the stimulatory action of heparin on muscle reinnervation and prevented the increase of insulin-like growth factor-I muscle levels.

Maeda, T., K. Kannari, et al. (2003). "Rapid induction of serotonergic hyperinnervation in the adult rat striatum with extensive dopaminergic denervation." Neurosci Lett 343(1): 17-20.
 The aim of our study was to determine whether serotonergic hyperinnervation is rapidly induced in the striatum of adult rats with extensive dopaminergic denervation. Immunohistochemical study was performed on the brain sections obtained at 2 and 8 weeks after injection of 6-hydroxydopamine (6-OHDA) into the medial forebrain bundle. The extent of dopaminergic denervation was evaluated as a percentage loss of tyrosine hydroxylase immunopositive neurons in the substantia nigra pars compacta. The immunopositive areas for serotonin (5-HT) in the striatum were measured. In the lesioned rats 97.5+/-0.6% of dopamine neurons were lost. 5-HT immunopositive areas in the striatum were significantly increased both at 2 and 8 weeks after 6-OHDA injection (P<0.01). These results suggest that extensive dopaminergic denervation in adult rats induces rapid serotonergic hyperinnervation in the striatum as early as 2 weeks after lesioning.

Maggi, S. P., J. B. Lowe, 3rd, et al. (2003). "Pathophysiology of nerve injury." Clin Plast Surg 30(2): 109-26.
 The response to nerve injury is a complex and often poorly understood mechanism. An in-depth and current command of the relevant neuroanatomy, classifications systems, and responses to injury and regeneration are critical to current clinical success. Continued progress must be made in our current understanding of these varied physiologic mechanisms of neuro-regeneration if any significant progress in clinical treatments or outcome is to be expected in the future. Reconstructive surgeons have in many ways maximized the technical aspects of peripheral nerve repair. However, advances in functional recovery may be seen with improvements in sensory and motor rehabilitation after peripheral nerve surgery and with a combined understanding of the neurobiology and neurophysiology of nerve injury and regeneration.

Magdalena, J., T. H. Millard, et al. (2003). "Microtubule involvement in NIH 3T3 Golgi and MTOC polarity establishment." J Cell Sci 116(Pt 4): 743-56.
 Scratch-wound assays are commonly used to study the ability of cells to polarize and migrate. In a previous study we showed that Golgi reorientation in response to a scratch wound is actin-dependent in NIH 3T3 cells but not in astrocytes. In this investigation, to study cell polarity and motility further, we used the polarization of the Golgi and microtubule organizing center (MTOC), as well as the ability of NIH 3T3 cells to migrate, in a scratch-wound assay. Unlike Golgi polarization, MTOC polarization was not dependent on actin, the Arp2/3 complex or Wiskott-Aldrich syndrome protein (WASP)-family proteins. By contrast, disruption of microtubules inhibited MTOC polarity, but not Golgi polarity. Migration was found to be dependent both on actin and microtubules. Expression of the formin-homology 2 (FH2) region of mDia1 inhibited Golgi polarization and migration but not MTOC polarization. Similarly, ST638, a Src inhibitor, inhibited Golgi polarization and migration but not MTOC polarization, whereas expression of the actin regulator IRSp53 only inhibited cell migration. Interestingly, the inhibition of cell migration by the mDia1 FH2 domain could be overcome by addition of Y27632, an inhibitor of ROCK (Rho-associated kinase). In fact, in the presence of ROCK inhibitor, cell migration was accelerated but polarization of both the Golgi and MTOC were inhibited. These data show that, in NIH 3T3 cells, different aspects of cell polarization and migration occur by different mechanisms, and both actin and microtubule networks are required. In addition, this study indicates that MTOC and Golgi polarization events are separately controlled.

Mahoney, S. A., R. Hosking, et al. (2003). "The second galanin receptor GalR2 plays a key role in neurite outgrowth from adult sensory neurons." J Neurosci 23(2): 416-21.
 Expression of the neuropeptide galanin is markedly upregulated within the adult dorsal root ganglion (DRG) after peripheral nerve injury. We demonstrated previously that the rate of peripheral nerve regeneration is reduced in galanin knock-out mice, with similar deficits observed in neurite outgrowth from cultured mutant DRG neurons. Here, we show that the addition of galanin peptide significantly enhanced neurite outgrowth from wild-type sensory neurons and fully rescued the observed deficits in mutant cultures. Furthermore, neurite outgrowth in wild-type cultures was reduced to levels observed in the mutants by the addition of the galanin antagonist M35 [galanin(1-13)bradykinin(2-9)]. Study of the first galanin receptor (GalR1) knock-out animals demonstrated no differences in neurite outgrowth compared with wild-type animals. Similarly, use of a GalR1-specific antagonist had no effect on neuritogenesis. In contrast, use of a GalR2-specific agonist had equipotent effects on neuritogenesis to galanin peptide, and inhibition of PKC reduced neurite outgrowth from wild-type sensory neurons to that observed in galanin knock-out cultures. These results demonstrate that adult sensory neurons are dependent, in part, on galanin for neurite extension and that this crucial physiological process is mediated by activation of the GalR2 receptor in a PKC-dependent manner.

Maki, H., M. Watanabe, et al. (2003). "Axons of alpha ganglion cells regenerate faster than other types into a peripheral nerve graft in adult cats." J Neurosci Res 72(2): 218-26.
 We reported previously that alpha ganglion cells in the cat retina have the highest ability in axonal regeneration. To examine whether alpha cells regenerate axons faster, we estimated the rate of axonal regeneration of retinal ganglion cells (RGCs) with transplantation of a peripheral nerve (PN) segment. After 4, 6, and 8 weeks of survival, regenerated RGCs were double-labeled with two fluorescent dyes injected separately at 10 mm and 20 mm from the connected site. From a scatter diagram of double-labeling ratios, we estimated that axons reached 20 mm by 3.2 weeks. Immunostaining suggested that first axon sprouts entered a PN segment on Day 4. These values enabled us to estimate average rates of axonal regeneration as 1.1 mm/day for all the RGCs. Proportions of cell types of regenerated RGCs were obtained with Lucifer yellow injections, and those of alpha cells were higher than those in normal retinas in any periods. From analysis of scatter diagrams, we estimated axonal growth rate of alpha, beta, and non-alpha/beta cells as 1.4, 1.1, 1.0 mm/day, respectively. The higher regeneration rate of alpha cells may reflect greater regenerative ability compared to other cell types. The present system also provides control values when a method to promote axonal regeneration is developed.

Malave, C., G. M. Villegas, et al. (2003). "Role of glypican-1 in the trophic activity on PC12 cells induced by cultured sciatic nerve conditioned medium: identification of a glypican-1-neuregulin complex." Brain Res 983(1-2): 74-83.
 Glypican-1 is an extracellular matrix component found by microsequencing in a medium conditioned by cultured rat-sciatic nerves (CM). This CM was concentrated by ultrafiltration and fractionated by quaternary ammonium chromatography, followed by Hi-Trap blue affinity chromatography to obtain the active fraction B1.2. Previously, we have reported a 54 kDa neuregulin (NRG) in the same B1.2 fraction [Villegas et al., Brain Res. 852 (2001) 304]. The effect of Glypican-1 on the neuron-like differentiation of PC12 cells was investigated by immunoprecipitation, Western blot and cellular image analysis. Removal of glypican-1 by immunoprecipitation with increasing concentrations of specific antibodies revealed a gradual decrease of the differentiation activity of fraction B1.2, which paralleled the results obtained by removal of the 54 kDa NRG protein. Colorless native electrophoresis and Western blot analysis was used to identify a glypican-1-NRG protein complex, which could be afterwards separated by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis into its individual components. Additionally, it was demonstrated that glypican-1, in cooperation with the 54 kDa NRG, is involved in the neuronal-like differentiation of PC12 cells and could play an important role on the regeneration responses of peripheral nerves.

Malik, R. A., P. Kallinikos, et al. (2003). "Corneal confocal microscopy: a non-invasive surrogate of nerve fibre damage and repair in diabetic patients." Diabetologia 46(5): 683-8.
 AIMS/HYPOTHESIS: The accurate detection, characterization and quantification of human diabetic neuropathy are important to define at risk patients, anticipate deterioration, and assess new therapies. Corneal confocal microscopy is a reiterative, rapid, non-invasive in vivo clinical examination technique capable of imaging corneal nerve fibres. The aim of this study was to define the ability of this technique to quantify the extent of degeneration and regeneration of corneal nerve fibres in diabetic patients with increasing neuropathic severity. METHODS: We scanned the cornea and collected images of Bowman's layer (containing a rich nerve plexus) from 18 diabetic patients and 18 age-matched control subjects. RESULTS: Corneal nerve fibre density (F(3)=9.6, p<0.0001), length (F(3)=23.8, p<0.0001), and branch density (F(3)=13.9, p<0.0001) were reduced in diabetic patients compared with control subjects, with a tendency for greater reduction in these measures with increasing severity of neuropathy. CONCLUSION/INTERPRETATION: Corneal confocal microscopy is a rapid, non-invasive in vivo clinical examination technique which accurately defines the extent of corneal nerve damage and repair and acts as a surrogate measure of somatic neuropathy in diabetic patients. It could represent an advance to define the severity of neuropathy and expedite assessment of therapeutic efficacy in clinical trials of human diabetic neuropathy.

Maniwa, S., A. Iwata, et al. (2003). "Effects of neurotrophic factors on chemokinesis of Schwann cells in culture." Scand J Plast Reconstr Surg Hand Surg 37(1): 14-7.
 Schwann cells are support cells in the peripheral nervous system and are responsible for migration, adhesion, production of the extracellular matrix, and myelination. Migration is considered to be essential for nerve regeneration after transsection. We have examined the chemokinetic effects of nerve growth factor (NGF), brain derived growth factor (BDNF), and neurotrophin-3 (NT-3) on Schwann cells in vitro using a chemotaxis chamber. The chemokinetic activity of Schwann cells was strongly accelerated by NGF, but was not influenced by BDNF. NT-3 at a concentration of 1 ng/ml had a stimulatory effect on chemokinesis. These data suggest that NGF is a chemoattractant for Schwann cells in vitro. Giving exogenous NGF might stimulate both migration of Schwann cells and the formation of Bungner's bands after peripheral nerve injuries in animal models.

Manni, L. and T. Lundeberg (2003). "Effects of cholecystokinin-8 in peripheral neuropathies: a nerve growth factor mediated action?" Arch Ital Biol 141(2-3): 117-26.
 
Marcol, W., K. Kotulska, et al. (2003). "Regeneration of sciatic nerves of adult rats induced by extracts from distal stumps of pre-degenerated peripheral nerves." J Neurosci Res 72(3): 417-24.
 Despite numerous experimental and clinical attempts to reconstruct injuries of peripheral nerves, the methods developed until now have not been sufficiently effective. We examined the influence of extracts (postmicrosomal fractions) obtained from non-pre-degenerated or 7-day-pre-degenerated distal segments of peripheral nerves on the regeneration of injured sciatic nerves of male adult rats. The extracts were introduced to the site of injury with autologous connective tissue chambers filled with fibrin. Reference groups were treated with brain-derived neurotrophic factor (BDNF) or fibrin only. We examined DiI-labeled motoneurons, toluidine blue-labeled myelinated fibers in the mid-part of the chambers, and AChE-positive nerve endings to assess the regeneration intensity. In addition, the length of fibers regrowing within the chambers was measured. We found that extracts obtained from distal stumps of 7-day-pre-degenerated peripheral nerves enhanced nerve regeneration as strongly as BDNF.

Marqueste, T., J. R. Alliez, et al. (2003). "Neuromuscular Rehabilitation by Treadmill Running or Electrical Stimulation after Peripheral Nerve Injury and Repair." J Appl Physiol.
 Numerous studies were devoted to the regeneration of the motor pathway toward a denervated muscle after nerve injury. However, the regeneration of sensory muscle endings after repair by self-anastomosis was few studied. In previous electrophysiological studies, we showed that the functional characteristics of tibialis anterior muscle afferents are differentially affected after injury and repair of the peroneal nerve with and without chronically electrostimulation. The present study focuses on the axonal regeneration of mechano- (fibers I and II) and metabosensitive (fibers III and IV) muscle afferents by evaluating the recovery of their response to different test agents after nerve injury, repair by self-anastomosis, during 10 weeks of treadmill running (LSR group). Data were compared to control animals (C), animals with nerve lesion and suture (LS), and animals with lesion, suture and chronic muscle rehabilitation by electrostimulation (LSE) with a biphasic current modulated in pulse duration and frequency, eliciting a pattern mimicking the activity delivered by the nerve to the muscle. Compared to the C group, results indicated that, 1) muscle weight was smaller in LS and LSR groups, 2) the fatigue index was greater in LS group and smaller in LSE group, 3) metabosensibility remained altered in the LS and LSE groups, and 4) mechanosensitivity presented a large increase of the activation pattern in the LS and LSE groups. Our data indicated that chronic muscle electrostimulation partially favors the recovery of muscle properties (i.e. muscle weight and twitch response were close to the controls) and that rehabilitation by treadmill running also efficiently induced a better functional muscle afferent recovery (i.e. the discharge pattern was similar to the controls). The effectiveness of the chronic electromyostimulation and the treadmill exercise on afferent recovery is discussed with regard to parameters listed above, is discussed.

Marsal, M., D. Pineda, et al. (2003). "Gtwnt-5 a member of the wnt family expressed in a subpopulation of the nervous system of the planarian Girardia tigrina." Gene Expr Patterns 3(4): 489-95.
 Wnt proteins are a family of highly conserved secreted glycoproteins that regulate cell-to-cell interactions during embryogenesis. They act as signaling molecules and take part in many crucial decisions throughout the development of organisms ranging from Hydra to human. We have isolated and characterized the expression of a member of the Wnt family, Gtwnt-5 gene in the planarian Girardia tigrina. Planarians are free-living members (Class Turbellaria) of the Phylum Platyhelminthes. They are best known for their high regenerative capabilities. These organisms have an apparently simple central nervous system (CNS) from a morphological perspective, with cephalic ganglia in the dorsal anterior region and two ventral main nerve cords along the body. However, a large number of planarian neural genes have recently been identified and therefore it is possible to define different molecular and functional domains in the planarian brain. The present study shows expression of Gtwnt-5 in a subpopulation of the whole CNS of intact organisms, being activated during regeneration. Gtwnt-5 reveals a differential spatial pattern: the expression is preferentially found in the most external region of the CNS. In addition, a kind of iterative pattern has been observed at the ganglia level, suggesting that the planarian brain might not be a continuous structure but compartmented or regionalized. Gtwnt-5 signal is also detected at the sensors of the worm: at the auricle level and all around the cephalic periphery. All these data provide us with a new neural marker for the planarian brain, and can be used to follow regeneration of the CNS.

Martinez-Gonzalez, J., J. Rius, et al. (2003). "Neuron-derived orphan receptor-1 (NOR-1) modulates vascular smooth muscle cell proliferation." Circ Res 92(1): 96-103.
 Vascular smooth muscle cells (VSMCs) migration and proliferation play a key role in the pathophysiology of cardiovascular disease. However, the transcription factors that regulate VSMC activation are not completely characterized. By a mRNA-differential display approach, we have identified neuron-derived orphan receptor-1 (NOR-1), a transcription factor within the NGFI-B subfamily of nuclear receptors, as a immediate-early gene in VSMCs. Two NOR-1 isoforms (alpha and beta) were identified and cloned from serum-induced porcine VSMC that shared high homology with the human isoforms. Northern blot analysis revealed a strong and transient (1 to 6 hours) upregulation of NOR-1 in both porcine and human coronary SMCs by growth factors (serum, platelet-derived growth factor-BB, and epidermal growth factor) and alpha-thrombin but not by cytokines. NOR-1 upregulation is processed through G protein-coupled receptors and tyrosine kinase receptors, and involves Ca2+ mobilization, protein kinase C activation, and the mitogen-activated protein kinase pathway. This induction was closely dependent of the cAMP response elements present in NOR-1 promoter as transfection assays indicate. Human coronary atherosclerotic lesions overexpress NOR-1, and balloon angioplasty transiently induces NOR-1 in porcine coronary arteries with a pattern similar to that observed in VSMCs in culture. Antisense oligonucleotides against NOR-1 inhibited human coronary SMC proliferation (reduced de novo DNA synthesis, cell cycle progression, and VSMC wound repair) as efficiently as antisense against the protooncogene c-fos. These results show that NOR-1 modulates VSMC proliferation, and suggest that this transcription factor may play a role in both spontaneous and accelerated atherosclerosis.

Mason, M. R., A. R. Lieberman, et al. (2003). "Corticospinal neurons up-regulate a range of growth-associated genes following intracortical, but not spinal, axotomy." Eur J Neurosci 18(4): 789-802.
 The failure of some CNS neurons to up-regulate growth-associated genes following axotomy may contribute to their failure to regenerate axons. We have studied gene expression in rat corticospinal neurons following either proximal (intracortical) or distal (spinal) axotomy. Corticospinal neurons were retrogradely labelled with cholera toxin subunit B prior to intracortical lesions or concomitantly with spinal lesions. Alternate sections of forebrain were immunoreacted for cholera toxin subunit B or processed for mRNA in situ hybridization for ATF3, c-jun, GAP-43, CAP-23, SCG10, L1, CHL1 or krox-24, each of which has been associated with axotomy or axon regeneration in other neurons. Seven days after intracortical axotomy, ATF3, c-jun, GAP-43, SCG10, L1 and CHL1, but not CAP-23 or krox-24, were up-regulated by layer V pyramidal neurons, including identified corticospinal neurons. The maximum distance between the lesion and the neuronal cell bodies that up-regulated genes varied between 300 and 500 microm. However, distal axotomy failed to elicit changes in gene expression in corticospinal neurons. No change in expression of any molecule was seen in the neocortex 1 or 7 days after corticospinal axotomy in the cervical spinal cord. The expression of GAP-43, CAP-23, L1, CHL1 and SCG10 was confirmed to be unaltered after this type of injury in identified retrogradely labelled corticospinal neurons. Thus, while corticospinal neuronal cell bodies fail to respond to spinal axotomy, these cells behave like regeneration-competent neurons, up-regulating a wide range of growth-associated molecules if axotomized within the cerebral cortex.

Mason, M. R., A. R. Lieberman, et al. (2003). "FKBP12 mRNA expression is upregulated by intrinsic CNS neurons regenerating axons into peripheral nerve grafts in the brain." Exp Neurol 181(2): 181-9.
 We have examined the expression of the immunophilin FKBP12 in adult rat intrinsic CNS neurons stimulated to regenerate axons by the implantation of segments of autologous tibial nerve into the thalamus or cerebellum. After survival times of 3 days to 6 weeks, the brains were fresh-frozen. In some animals the regenerating neurons were retrogradely labelled with cholera toxin subunit B 1 day before they were killed. Sections through the thalamus or cerebellum were used for in situ hybridization with digoxygenin-labelled riboprobes for FKBP12 or immunohistochemistry to detect cholera toxin subunit B-labelled neurons. FKBP12 was constitutively expressed by many neurons, and was very strongly expressed in the hippocampus and by Purkinje cells. Regenerating neurons were found in the thalamic reticular nucleus and deep cerebellar nuclei of animals that received living grafts. Neurons in these nuclei upregulated FKBP12 mRNA; such neurons were most numerous at 3 days post grafting but were most strongly labelled at 2 weeks post grafting. Regenerating neurons identified by retrograde labelling were found to have upregulated FKBP12 mRNA. No upregulation was seen in neurons in animals that received freeze-killed grafts, which do not support axonal regeneration. We conclude that FKBP12 is a regeneration-associated gene in intrinsic CNS neurons.

Matsumoto, N., H. Kitayama, et al. (2003). "Isolation of a set of genes expressed in the choroid plexus of the mouse using suppression subtractive hybridization." Neuroscience 117(2): 405-15.
 The choroid plexus produces cerebrospinal fluid, providing a specialized environment for the CNS. We previously demonstrated that choroid plexus ependymal cells can enhance nerve regeneration in vivo and promote neurite outgrowth in vitro. To understand the molecular mechanisms of choroid plexus functions, we isolated genes predominantly expressed in the mouse choroid plexus using suppression subtractive hybridization. Out of the 49 complementary DNA (cDNA) fragments isolated in two types of screening, 43 matched known sequences in the database and six were novel. In one type of screening where choroid plexus cDNAs were subtracted with cerebral cortex cDNAs, transthyretin and phosphodiesterase I alpha were predominant. This is consistent with previous reports and supports the authenticity of our approach. In the other type of screening, cDNAs derived from the choroid plexus of neonatal (postnatal day 5) mice were subtracted with cDNAs from the choroid plexus of adult mice. RNA blot and/or in situ hybridization confirmed abundant expression, in the mouse choroid plexus, of the mRNA encoding gelsolin, phospholipid transfer protein, ATP-binding cassette transporter A8 (ABCA8), androgen-inducible aldehyde reductase, and Na(+)/sulfate cotransporter SUT-1. Also, one novel gene (FS88) was found to be expressed in the choroid plexus from neonatal mice. Our data suggest that the choroid plexus cells produce molecules involved in processes such as prevention of fibrillization of amyloid beta-protein (transthyretin and gelsolin), lipid metabolism (phospholipid transfer protein and ABCA8), and detoxification (androgen-inducible aldehyde reductase).

Matsuo, E. (2003). "[The sequelae of Hansen's disease. (Pathologic viewpoint of etiologies, morphologies and countermeasures)]." Nihon Hansenbyo Gakkai Zasshi 72(3): 251-7.
 The proportion of glomerulonephritis, often a sequence of arteriolitis, among the sequelae of Hansen's disease after the introduction of chemotherapy increased markedly in Japan and nullified that of once prevalent tuberculosis after 1960s. However, most significant aftermath of the disease for numbers of years in the past have been peripheral nerve injuries worldwide for which effective countermeasures are yet to be developed. In this brief autopsy cases study from 1960s to 1990s, we confirmed the presence of cases in which arteriolitis and resulted infarction of peripheral nerves and not M. leprae itself were shown to be the major cause of axonal damages. There were also cases in which the accumulation of the bacilli without vascular changes did not damage the axons. The cases as these could not be solitary but should be rather common in this time of chemotherapy. If so, the methods to reconstruct nerves and blood vessels by promoting those regeneration should be developed to cope with the situation for surgeon, assisted by pathologists.

McDonald, J. W. and D. Becker (2003). "Spinal cord injury: promising interventions and realistic goals." Am J Phys Med Rehabil 82(10 Suppl): S38-49.
 Long regarded as impossible, spinal cord repair is approaching the realm of reality as efforts to bridge the gap between bench and bedside point to novel approaches to treatment. It is important to recognize that the research playing field is rapidly changing and that new mechanisms of resource development are required to effectively make the transition from basic science discoveries to effective clinical treatments. This article reviews recent laboratory studies and phase 1 clinical trials in neural and nonneural cell transplantation, stressing that the transition from basic science to clinical applications requires a parallel rather than serial approach, with continuous, two-way feedback to most efficiently translate basic science findings, through evaluation and optimization, to clinical treatments. An example of mobilizing endogenous stem cells for repair is reviewed, with emphasis on the rapid application of basic science to clinical therapy. Successful and efficient transition from basic science to clinical applications requires (1) a parallel rather than a serial approach; (2) development of centers that integrate three spheres of science, translational, transitional, and clinical trials; and (3) development of novel resources to fund the most critically limited step of transitional to clinical trials.

McDonald, D. S. and D. W. Zochodne (2003). "An injectable nerve regeneration chamber for studies of unstable soluble growth factors." J Neurosci Methods 122(2): 171-8.
 Modern surgical techniques cannot guarantee functional recovery following peripheral nerve injuries. Research into factors that may influence nerve regeneration has therefore assumed a prominent potential therapeutic role. We report here on the development of an approach to allow for direct manipulation of the microenvironment of regenerating peripheral nerve axons. We show that solutions can be delivered directly to this local milieu in vivo and that such a delivery can be performed multiple times over an extended period, potentially facilitating studies of multiple molecular players that act locally. We also demonstrate that the bundle of regenerated axons are amenable to morphological analysis by 21 days and that the injection system remains patent for at least 21 days.

McGee, A. W. and S. M. Strittmatter (2003). "The Nogo-66 receptor: focusing myelin inhibition of axon regeneration." Trends Neurosci 26(4): 193-8.
 CNS myelin inhibits axonal outgrowth in vitro and is one of several obstacles to functional recovery following spinal cord injury. Central to our current understanding of myelin-mediated inhibition are the membrane protein Nogo and the Nogo-66 receptor (NgR). New findings implicate NgR as a point of convergence in signal transduction for several myelin-associated inhibitors. Additional studies have identified a potential coreceptor for NgR as p75(NTR), and a second-messenger pathway involving RhoA that inhibits neurite elongation. Although these findings expand our understanding of the molecular determinants of adult CNS axonal regrowth, the physiological roles of myelin-associated inhibitors in the intact adult CNS remain ill-defined.

McKay Hart, A., M. Wiberg, et al. (2003). "Exogenous leukaemia inhibitory factor enhances nerve regeneration after late secondary repair using a bioartificial nerve conduit." Br J Plast Surg 56(5): 444-50.
 The clinical outcome of peripheral nerve injuries remains disappointing, even in the ideal situation of a primary repair performed with optimal microsurgical techniques. Primary repair is appropriate for only about 85% of injuries, and outcome is worse following secondary nerve repair, partly owing to the reduced regenerative potential of chronically axotomised neurons. Leukaemia inhibitory factor (LIF) is a gp-130 neurocytokine that is thought to act as an 'injury factor', triggering the early-injury phenotype within neurons and potentially boosting their regenerative potential after secondary nerve repair. At 2-4 months after sciatic nerve axotomy in the rat, 1 cm gaps were repaired using either nerve isografts or poly-3-hydroxybutyrate conduits containing a calcium alginate and fibronectin hydrogel.Regeneration was determined by quantitative immunohistochemistry 6 weeks after repair, and the effect of incorporating recombinant LIF (100 ng/ml) into the conduits was assessed. LIF increased the regeneration distance in repairs performed after both 2 months (69%, P=0.019) and 4 months (123%, P=0.021), and was statistically comparable to nerve graft. The total area of axonal immunostaining increased by 21% (P>0.05) and 63% (P>0.05), respectively. Percentage immunostaining area was not increased in the 2 months group, but increased by 93% in the repairs performed 4 months after axotomy. Exogenous LIF, therefore, has a potential role in promoting peripheral nerve regeneration after secondary repair, and can be effectively delivered within poly-3-hydroxybutyrate bioartificial conduits used for nerve repair.

McMurray, R., R. Islamov, et al. (2003). "Raloxifene analog LY117018 enhances the regeneration of sciatic nerve in ovariectomized female mice." Brain Res 980(1): 140-5.
 The aim of this study was to examine the effects of LY117018, a selective estrogen receptor modulator, on peripheral nerve regeneration, using a model of sciatic nerve crush injury in mice. Sciatic functional index, an index of functional recovery, was significantly higher in LY117018 treated mice throughout regeneration. Analysis of semi-thin sections revealed a significant increase in both the total number of regenerating nerve fibers at day 7, and the mean axonal area of myelinated fibers at 7, 14, and 21 days after injury, in LY117018 treated mice. Analysis of axonal transport through retrograde labeling of motor neurons showed that LY117018 increased transport, and ICI 182,780 blocked the effects of LY117018, delineating estrogen receptors as its target. Our study suggests that LY117018 may markedly accelerate peripheral nerve regeneration and functional recovery through activation of estrogen receptors.

Mears, S., M. Schachner, et al. (2003). "Antibodies to myelin-associated glycoprotein accelerate preferential motor reinnervation." J Peripher Nerv Syst 8(2): 91-9.
 Predegeneration of nerve enhances its ability to support axon regeneration. Trophic factors are upregulated by reactive Schwann cells while potentially inhibitory molecules are removed. These experiments isolate the effects of one such inhibitory molecule, the myelin-associated glycoprotein (MAG), to determine its role in modifying regeneration after nerve repair. Suture of the mouse femoral nerve was followed by daily intraperitoneal injection of antibodies to MAG, antibodies to HNK-1, a specific muscle pathway marker, or no further treatment. Regeneration was assayed by double-labeling the femoral cutaneous and muscle branches with horseradish peroxidase and fluoro-gold after 4 weeks or 6 weeks of regeneration. Four weeks after nerve repair, selective reinnervation of the muscle branch by motoneurons, or preferential motor reinnervation (PMR), was not seen in either controls or L2-antibody-treated animals. In contrast, treatment with MAG antibodies resulted in dramatic PMR. By 6 weeks, the controls had achieved borderline specificity, substantial PMR developed in the L2 antibody group and the MAG group changed little. Blocking access to MAG in the distal nerve stump thus accelerated and enhanced PMR. Sensory regeneration was depressed by both antibody treatments at 4 weeks but recovered by 6 weeks. Antibody administration has a generalized effect on sensory regeneration that is unrelated to the behavior of motoneurons in the same nerve.

Meek, M. F. and J. Henk Coert (2003). "Turnover epineural sheath tube in primary repair of peripheral nerves." Ann Plast Surg 50(3): 328-30; author reply 330.
 
Meek, M. F., M. J. Bertleff, et al. (2003). "[A degradable artificial nerve guide to bridge peripheral nerve defects]." Ned Tijdschr Geneeskd 147(15): 717-21.
 The standard technique for bridging a peripheral nerve defect is an autologous nerve graft if the nerve ends cannot be sutured. Recent evidence indicates that an alternative procedure-application of a degradable nerve guide-may be feasible. Currently the use of such a degradable nerve guide for the recovery of peripheral nerve defects in the hand is being tested in a multicenter trial. Conventional suturing as well as autologous nerve grafting are accepted methods in the control group within the protocol of the multicenter study. In the first two patients to receive the implant, a 28-year-old man and a 50-year-old women with tendon and nerve injury due to glass cuts, the operation was technically successful. Functional nerve recovery will be assessed in due course.

Meek, M. F., J. H. Coert, et al. (2003). "Recovery of touch after median nerve lesion and subsequent repair." Microsurgery 23(1): 2-5.
 Many techniques have been developed for the evaluation of peripheral nerve function. Consequently, physicians use different techniques in the clinic. This study describes the evaluation of touch after median nerve lesions in the forearm and repair. In order to evaluate touch, 25 patients, aged 11-51 years (mean, 29 years), were evaluated 3-10.5 years (mean, 5 years) after median nerve repair. The evaluation included the moving two-point discrimination test and Semmes-Weinstein monofilaments. We showed that 32% good-excellent results can be obtained with difficult nerve lesions. The results could have been improved if a sensory reeducation regime had been applied.

Meiners, S. and M. L. Mercado (2003). "Functional peptide sequences derived from extracellular matrix glycoproteins and their receptors: strategies to improve neuronal regeneration." Mol Neurobiol 27(2): 177-96.
 Peptides derived from extracellular matrix proteins have the potential to function as potent therapeutic reagents to increase neuronal regeneration following central nervous system (CNS) injury, yet their efficacy as pharmaceutical reagents is dependent upon the expression of cognate receptors in the target tissue. This type of codependency is clearly observed in successful models of axonal regeneration in the peripheral nervous system, but not in the normally nonregenerating adult CNS. Successful regeneration is most closely correlated with the induction of integrins on the surface of peripheral neurons. This suggests that in order to achieve optimal neurite regrowth in the injured adult CNS, therapeutic strategies must include approaches that increase the number of integrins and other key receptors in damaged central neurons, as well as provide the appropriate growth-promoting peptides in a "regeneration cocktail." In this review, we describe the ability of peptides derived from tenascin- C, fibronectin, and laminin-1 to influence neuronal growth. In addition, we also discuss the implications of peptide/receptor interactions for strategies to improve neuronal regeneration.

Mendonca, A. C., C. H. Barbieri, et al. (2003). "Directly applied low intensity direct electric current enhances peripheral nerve regeneration in rats." J Neurosci Methods 129(2): 183-90.
 The influence of direct electric stimulation on nerve regeneration was studied in a model of crush injury of the sciatic nerve of rats. Forty-three rats were used and distributed in four groups according to the procedure: (1) intact nerve, inactive circuit; (2) crush injury, inactive circuit; (3) intact nerve, active circuit; (4) crush injury, active circuit. The low intensity continuous current circuit (1 microA) was implanted in the lumbar region, the anode being fixed to the muscles proximally and the cathode below the nerve distally to the lesion site. The Sciatic Functional Index (SFI) was evaluated at weekly intervals for 3 weeks, the sciatic nerve being resected on the 21st day for histologic and morphometric studies. The SFI progressively improved and the average fiber nerve density recovered to a nearly normal value in Group 2 and increased in Group 4 compared with the control groups (1 and 3), but this was accompanied by a decreased average fiber nerve diameter. Both number and diameter of inter and intra-fascicular blood vessels increased in the stimulated nerves. We conclude that low intensity direct electric stimulation enhances nerve regeneration following a controlled nerve crush injury and increases blood supply by increasing number and diameter of vasa nervorum.

Mendez-Otero, R. and L. A. Cavalcante (2003). "Functional role of gangliosides in neuronal motility." Prog Mol Subcell Biol 32: 97-124.
 
Menet, V., M. Prieto, et al. (2003). "Axonal plasticity and functional recovery after spinal cord injury in mice deficient in both glial fibrillary acidic protein and vimentin genes." Proc Natl Acad Sci U S A 100(15): 8999-9004.
 The lack of axonal regeneration in the injured adult mammalian spinal cord leads to permanent functional disabilities. The inability of neurons to regenerate their axon is appreciably due to an inhospitable environment made of an astrocytic scar. We generated mice knock-out for glial fibrillary acidic protein and vimentin, the major proteins of the astrocyte cytoskeleton, which are upregulated in reactive astrocytes. These animals, after a hemisection of the spinal cord, presented reduced astroglial reactivity associated with increased plastic sprouting of supraspinal axons, including the reconstruction of circuits leading to functional restoration. Therefore, improved anatomical and functional recovery in the absence of both proteins highlights the pivotal role of reactive astrocytes in axonal regenerative failure in adult CNS and could lead to new therapies of spinal cord lesions.

Menovsky, T. and J. F. Beek (2003). "Carbon dioxide laser-assisted nerve repair: effect of solder and suture material on nerve regeneration in rat sciatic nerve." Microsurgery 23(2): 109-16.
 In order to further improve and explore the role of lasers for nerve reconstruction, this study was designed to investigate regeneration of sharply transected peripheral nerves repaired with a CO(2) milliwatt laser in combination with three different suture materials and a bovine albumin protein solder as an adjunct to the welding process. Unilateral sciatic nerve repair was performed in 44 rats. In the laser group, nerves were gently apposed, and two stay sutures (10-0 nylon, 10-0 polyglycolic acid, or 25 microm stainless steel) were placed epi/perineurially. Thereafter, the repair site was fused at 100 mW with pulses of 1.0 s. In the subgroup of laser-assisted nerve repair (LANR), albumen was used as a soldering agent to further reinforce the repair site. The control group consisted of nerves repaired by conventional microsurgical suture repair (CMSR), using 4-6 10-0 nylon sutures. Evaluation was performed at 1 and 6 weeks after surgery, and included qualitative and semiquantitative light microscopy. LANR performed with a protein solder results in a good early peripheral nerve regeneration, with an optimal alignment of nerve fibers and minimal connective tissue proliferation at the repair site. All three suture materials produced a foreign body reaction; the least severe was with polyglycolic acid sutures. CMSR resulted in more pronounced foreign-body granulomas at the repair site, with more connective-tissue proliferation and axonal misalignment. Furthermore, axonal regeneration in the distal nerve segment was better in the laser groups. Based on these results, CO(2) laser-assisted nerve repair with soldering in combination with absorbable sutures has the potential of allowing healing to occur with the least foreign-body reaction at the repair site. Further experiments using this combination are in progress.

Midha, R., C. A. Munro, et al. (2003). "Growth factor enhancement of peripheral nerve regeneration through a novel synthetic hydrogel tube." J Neurosurg 99(3): 555-65.
 OBJECT: The authors' long-term goal is repair of peripheral nerve injuries by using synthetic nerve guidance devices that improve both regeneration and functional outcome relative to an autograft. They report the in vitro processing and in vivo application of synthetic hydrogel tubes that are filled with collagen gel impregnated with growth factors. METHODS: Poly(2-hydroxyethyl methacrylate-co-methyl methacrylate) (PHEMA-MMA) porous 12-mm-long tubes with an inner diameter of 1.3 mm and an outer diameter of 1.8 mm were used to repair surgically created 10-mm gaps in the rat sciatic nerve. The inner lumen of the tubes was filled with collagen matrix alone or matrix supplemented with either neurotropin-3 at 1 microg/ml, brain-derived neurotrophic factor at 1 microg/ml, or acidic fibroblast growth factor (FGF-1) at 1 or 10 microg/ml. Nerve regeneration through the growth factor-enhanced tubes was assessed at 8 weeks after repair by histomorphometric analysis at the midgraft level and in the nerve distal to the tube repair. The tubes were biostable and biocompatible, and supported nerve regeneration in more than 90% of cases. Nerve regeneration was improved in tubes in which growth factors were added, compared with empty tubes and those containing collagen gel alone (negative controls). Tubes filled with 10 microg/ml of FGF-1 dispersed in collagen demonstrated regeneration comparable to autografts (positive controls) and showed significantly better regeneration than the other groups. CONCLUSIONS: The PHEMA-MMA tubes augmented with FGF-1 in their lumens appear to be a promising alternative to autografts for repair of nerve injuries. Studies are in progress to assess the long-term biocompatibility of these implants and to enhance regeneration further.

Miller, S. J., H. Li, et al. (2003). "Brain lipid binding protein in axon-Schwann cell interactions and peripheral nerve tumorigenesis." Mol Cell Biol 23(6): 2213-24.
 Loss of axonal contact characterizes Schwann cells in benign and malignant peripheral nerve sheath tumors (MPNST) from neurofibromatosis type 1 (NF1) patients. Tumor Schwann cells demonstrate NF1 mutations, elevated Ras activity, and aberrant epidermal growth factor receptor (EGFR) expression. Using cDNA microarrays, we found that brain lipid binding protein (BLBP) is elevated in an EGFR-positive subpopulation of Nf1 mutant mouse Schwann cells (Nf1(-/-) TXF) that grows away from axons; BLBP expression was not affected by farnesyltransferase inhibitor, an inhibitor of H-Ras. BLBP was also detected in EGFR-positive cell lines derived from Nf1:p53 double mutant mice and human MPNST. BLBP expression was induced in normal Schwann cells following transfection with EGFR but not H-Ras12V. Furthermore, EGFR-mediated BLBP expression was not inhibited by dominant-negative H-Ras, indicating that BLBP expression is downstream of Ras-independent EGFR signaling. BLBP-blocking antibodies enabled process outgrowth from Nf1(-/-) TXF cells and restored interaction with axons, without affecting cell proliferation or migration. Following injury, BLBP expression was induced in normal sciatic nerves when nonmyelinating Schwann cells remodeled their processes. These data suggest that BLBP, stimulated by Ras-independent pathways, regulates Schwann cell-axon interactions in normal peripheral nerve and peripheral nerve tumors.

Mizusawa, I., S. Abe, et al. (2003). "Expression of cytokines, neurotrophins, neurotrophin receptors and NOS mRNA in dorsal root ganglion of a rat tourniquet model." Leg Med (Tokyo) 5 Suppl 1: S271-4.
 We studied temporal changes in mRNA expression patterns for nitric oxide synthase (NOS), cytokines, neurotrophins and neurotrophin receptors in the dorsal root ganglion (DRG) of the rat, after application of a tourniquet to the hind limb. Collapsed myelin and degenerated axons were observed in the tourniquet segment of the sciatic nerve. Gene expression level of inducible nitric oxide synthase (iNOS) and neuronal nitric oxide synthase (nNOS) was significantly increased in ipsilateral DRG samples at 4h after application of the tourniquet but not in the contralateral or control DRG samples. Upregulation of tumor necrosis factor (TNF)-alpha, activating transcription factor (ATF)-3 and neurotrophin-3 (NT3) expressions began at 1h after application of the tourniquet in ipsilateral DRGs. It is likely that transient expression of these molecules triggers secondary events that may be beneficial to wound repair and regeneration.

Mligiliche, N. L., Y. Tabata, et al. (2003). "Poly lactic acid--caprolactone copolymer tube with a denatured skeletal muscle segment inside as a guide for peripheral nerve regeneration: a morphological and electrophysiological evaluation of the regenerated nerves." Anat Sci Int 78(3): 156-61.
 A biodegradable copolymer of poly L-lactic acid and epsilon-caprolactone (PLAC) was manufactured into a tube, in which a denatured skeletal muscle segment was placed longitudinally. This model tube was implanted as a guide to promote nerve regeneration across a 5 cm gap in the rabbit sciatic nerve. Five months after implantation, good nerve regeneration was found throughout the graft and in the distal host nerve. The population (29.6/16 x 10(2) microm(2)) of regenerated nerves in the graft was higher than that of the contralateral normal sciatic nerve (18.0/16 x 10(2) microm(2)). Regenerated nerve fibers extended to the distal host nerve. The number of myelinated fibers was 13.7/16 x 10(2) microm(2) at a level 1.5 cm from the distal suture. The diameters (below 2 microm) of most regenerated myelinated (nerves in the graft and in the distal host nerve were much smaller than those (6-8 microm) of normal nerves. Electrophysiological evaluation showed that the hindlimb muscle (gastrocnemius) was innervated by motor nerves in all animals 5 months after implantation. These results indicate that the PLAC tube with a denatured muscle segment inside provided good conditions for nerve fiber regrowth. The PLAC tube is thought to protect the denatured muscle segment from rapid dissociation in the host tissue.

Moilanen, J. A., M. H. Vesaluoma, et al. (2003). "Long-term corneal morphology after PRK by in vivo confocal microscopy." Invest Ophthalmol Vis Sci 44(3): 1064-9.
 PURPOSE: To examine human corneal morphology and nerve recovery 5 years after photorefractive keratectomy (PRK). METHODS: Fourteen eyes of 14 patients (ages, 27-53 years) who underwent 6-mm diameter PRK for low to moderate myopia (spherical equivalent [SE] -2.5 to -8.0 D) were examined once 5 years after surgery. Nine healthy individuals served as control subjects. Standard biomicroscopy, manifest refraction, and visual acuity tests were performed. The morphology of the corneas was examined by in vivo confocal microscope. Thicknesses of the epithelium and stroma, as well as the density of corneal opacity (haze) were obtained from digital image analysis of the confocal microscopy through-focusing (CMTF) scans. RESULTS: Confocal microscopy revealed increased reflectivity in the subepithelial extracellular matrix, keratocyte nuclei and processes in all patients. The mean objective haze estimate was 166.7 U (range, 50-390) in control corneas compared with a mean of 225.9 U (range, 125-430, P = 0.15) in the post-PRK corneas. The density of the subbasal nerve fiber bundles in post-PRK corneas (mean, n = 4.2; range, n = 1-7 per field of view) was not significantly lowered from that in control subjects (mean, n = 4.9; range, n = 3-6; P = 0.56). Bowman's layer was undetectable in all post-PRK corneas. Clinically, slit-lamp-observed trace of haze in four corneas correlated positively with the ablation depth (P = 0.016) and the thickness of the haze area (P = 0.006) in the confocal microscope. CONCLUSIONS: In vivo confocal microscopy demonstrates the presence of morphologic alterations even 5 years after PRK. However, these alterations are overcome by cellular and neural recovery and do not seem to interfere with visual performance.

Molski, M. (2003). "[First dorsal metacarpal flap in compound multi tissues thumb reconstruction]." Chir Narzadow Ruchu Ortop Pol 68(2): 115-9.
 In the years 1985-2000 two females and eleven males aged 3-48 (mean age 27) were treated using neurovascular first dorsal metacarpal flap because of compound thumb multi-tissue defects. In two cases the flap included a fragment of vascularized bone from the index proximal phalanx. Simultaneously a graft of non-vascularized bone in three cases, nerve grafts in two cases and a reconstruction of A1 pulley of the FPL tendon in three cases were carried out. Two patients were operated immediately after the injury and the others had delayed reconstructions. In 6 cases the defects resulted from crushing or lacerated wounds of the proximal thumb and in three cases of the distal thumb. Three thumbs were distally amputated. A 3 year-old child had an extensive palmar tissues necrosis due to an electric burn. Pedicled flaps were used in twelve patients and one patient had an island flap. The flaps ranged from 2.24 to 12.0 square centimetres. The donor wounds were directly closed in 6 patients and in 7 patients they were skin grafted. All flaps survived completely resulting in good function. Vascularized bone grafts raised in the flaps healed after 6 and 10 weeks respectively, those non-vascularized from the olecranon healed after 8 weeks and all were further rebuilt giving strong thumb of proper length. 2PD was 8-10 mm. Reconstruction of the sheath elements with flap tissues successfully supported FPL tendon. None of the donor site were observed.

Monnier, P. P., A. Sierra, et al. (2003). "The Rho/ROCK pathway mediates neurite growth-inhibitory activity associated with the chondroitin sulfate proteoglycans of the CNS glial scar." Mol Cell Neurosci 22(3): 319-30.
 Axons fail to regenerate in the central nervous system after injury. Chondroitin sulfate proteoglycans (CSPG) expressed in the scar significantly contribute to the nonpermissive properties of the central nervous system environment. To examine the inhibitory activity of a CSPG mixture on retina ganglion cell (RGC) axon growth, we employed both a stripe assay and a nerve fiber outgrowth assay. We show that the inhibition exerted by CSPGs in vitro can be blocked by application of either C3 transferase, a specific inhibitor of the Rho GTPase, or Y27632, a specific inhibitor of the Rho kinase. These results demonstrate that CSPG-associated inhibition of neurite outgrowth is mediated by the Rho/ROCK signaling pathway. Consistent with these results, we found that retina ganglion cell axon growth on glial scar tissue was enhanced in the presence of C3 transferase and Y27632, respectively. In addition, we show that the recently identified inhibitory CSPG Te38 is upregulated in the lesioned spinal cord.

Moon, L. D., R. A. Asher, et al. (2003). "Limited growth of severed CNS axons after treatment of adult rat brain with hyaluronidase." J Neurosci Res 71(1): 23-37.
 Many chondroitin sulfate proteoglycans (CSPGs) have been shown to influence CNS axon growth in vitro and in vivo. These interactions can be mediated through the core protein or through the chondroitin sulfate (CS) glycosaminoglycan (GAG) side chains. We have shown previously that degrading CS GAG side chains using chondroitinase ABC enhances dopaminergic nigrostriatal axon regeneration in vivo. We test the hypothesis that interfering with complete CSPGs also limit axon growth in vivo. Neurocan, versican, aggrecan, and brevican CSPGs may be anchored within extracellular matrix through binding to hyaluronan glycosaminoglycan. We examine whether degradation of hyaluronan using hyaluronidase might release these inhibitory CSPGs from the extracellular matrix and thereby enhance regeneration of cut nigrostriatal axons. Anesthetized adult rats were given knife cut lesions of the right hemisphere nigrostriatal tract and cannulae were secured transcranially thereby allowing repeated perilesional infusion of saline or saline containing hyaluronidase once daily for 10 days post-axotomy. Eleven days post-transection brains from animals under terminal anesthesia were recovered for histological evaluation. Effective delivery of substance was inferred from the observed reduction in perilesional immunoreactivity for neurocan and versican after treatment with hyaluronidase (relative to saline). Immunolabeling using antibodies against tyrosine hydroxylase was used to examine the response of cut dopaminergic nigral neurons. After transection and treatment with saline, dopaminergic nigral neurons sprouted in a region lacking astrocytes, neurocan and versican. Axons did not regenerate into the lesion surround that contained astrocytes and abundant neurocan and versican. After transection and treatment with hyaluronidase, there was a significant increase in the number of cut dopaminergic nigral axons growing up to 800 microm anterior to the site of transection. However, cut dopaminergic nigral axons still did not regenerate into the lesion surround that contained reduced (albeit residual) neurocan and versican immunoreactivity. Thus, partial degradation of hyaluronan and chondroitin sulfate and depletion of hyaluronan-binding CSPGs enhances local sprouting of cut CNS axons, but long-distance regeneration fails in regions containing residual hyaluronan-binding CSPGs. Hyaluronan, chondroitin sulfate and hyaluronan-binding CSPGs therefore likely contribute toward the failure of spontaneous axon regeneration in the injured adult mammalian brain and spinal cord.

Morcos, Y., S. M. Lee, et al. (2003). "A role for hypertrophic astrocytes and astrocyte precursors in a case of rapidly progressive multiple sclerosis." Mult Scler 9(4): 332-41.
 The purpose of this study was to examine the roles played by astrocytes in a case of rapidly progressive multiple sclerosis (MS). Within early-active and active lesions, hypertrophic astrocytes played an important role in lesion pathology through the phagocytosis of myelin and axonal debris and through the internalization of other glial cells, including astrocytes. In addition to this critical role, hypertrophic astrocytes, in areas that lack significant inflammation (within the adjacent normal appearing white matter and within late remyelinating lesions) were found to be active in myelin and axonal debris phagocytosis with no evidence of cellular internalization. Hypertrophic astrocytes therefore not only play an important role in the pathogenesis of MS lesions but also exert a continued deleterious effect upon tissue in the absence of significant inflammation. In addition, we found evidence for a significant population of vimentin-positive, glial fibrillary acidic protein (GFAP)-negative, bipolar, astrocyte precursors within the late remyelinating lesions. Their significance is not known but a possible role may include their participation in the successful remyelination of the lesion.

Moreno-Flores, M. T., F. Lim, et al. (2003). "Immortalized olfactory ensheathing glia promote axonal regeneration of rat retinal ganglion neurons." J Neurochem 85(4): 861-71.
 Olfactory bulb ensheathing glia (OEG) have attracted special attention during the last few years because of their unique properties in promoting regeneration of adult mammalian central nervous system (CNS) components. However the molecular and cellular characteristics responsible for this capacity remain to be revealed. Such studies are presently hindered by the lack of a plentiful source of homogenous OEG. Thus the availability of immortalized OEG lines maintaining the regenerative characteristics of the primary cultures would represent an unlimited source of OEG for use not only in biochemical analyses of neuroregenerative mechanisms but also to characterize their regenerative properties in models in culture and in vivo. We have immortalized primary rat OEG using the SV40 large T antigen expressed from a constitutive cellular promotor, and report here the isolation and characterization of clonal lines. These OEG clonal lines were comparable to primary OEG and Schwann cells in the promotion of axonal regeneration of mature rat retinal ganglion neurons (RGN) but, significantly, this culture assay system more closely reflects the in vivo reparative properties of OEG on transected nerves than other assays of neuritogenesis in that it revealed OEG cells to promote the growth of a larger number of long axons than Schwann cells. Using this assay we were able to grade our OEG lines for their neuroregenerative capacity, opening the possibility of identifying molecules with correlative expression levels in these cells. Our preliminary characterization revealed that the expression level of a classical OEG marker, the p75-NGF receptor, does not correlate with neuroregenerative capacity.

Morgenstern, D. A., R. A. Asher, et al. (2003). "Expression and glycanation of the NG2 proteoglycan in developing, adult, and damaged peripheral nerve." Mol Cell Neurosci 24(3): 787-802.
 We have investigated expression of the axon growth-inhibitory proteoglycan NG2 in peripheral nerve. In the adult, NG2 was present on endoneurial and perineurial fibroblasts, but not on Schwann cells. At birth, peripheral nerve NG2 was heavily glycanated, but was much less so in the adult. In vitro, sciatic nerve fibroblasts also produced heavily glycanated NG2. After peripheral nerve injury in rats and humans, an accumulation of NG2-positive cells was observed at the injury site. In the rat, there was an increase in NG2 glycanation for at least 2 weeks following injury. In mixed cultures of Schwann cells and peripheral nerve fibroblasts, the axons preferred to grow on the Schwann cells and seldom crossed onto the fibroblasts. Three-dimensional cultures of sciatic nerve fibroblasts were inhibitory to the growth of dorsal root ganglion axons. Inhibition of proteoglycan synthesis made the cells more permissive. NG2 may play a part in blocking axon regeneration through scar tissue in injured human peripheral nerve.

Mosahebi, A., M. Wiberg, et al. (2003). "Addition of fibronectin to alginate matrix improves peripheral nerve regeneration in tissue-engineered conduits." Tissue Eng 9(2): 209-18.
 Schwann cell (SC) transplantation has been proposed to encourage peripheral nerve regeneration, but an optimal SC-carrying matrix would be needed. The aim of this study was to characterize how the addition of fibronectin to alginate would affect the outcome of nerve regeneration promoted by Schwann cells embedded in this matrix. Genetically labeled rat SCs were obtained by lacZ gene transduction. SCs were suspended in alginate hydrogel matrix with/without addition of liquid fibronectin, and their viability and growth in the different types of matrices were assessed in vitro by AlamarBlue assay. In vivo assessment of SC transplantation in the matrix was carried out with poly-3-hydroxybutyrate (PHB) conduits to bridge a sciatic nerve gap. The grafted conduits were harvested at 2, 3, and 6 weeks and assessed for the presence of labeled SCs in relation to regrowing axons. The amount and rate of axonal regeneration were assessed by quantitative immunohistochemistry. Addition of fibronectin to alginate hydrogel improved SC viability and growth profile in vitro. X-Gal staining confirmed that SCs transplanted in PHB conduits were viable throughout the time course, and that the labeled SCs were clearly associated with regenerating axons. The regeneration rate was enhanced when liquid fibronectin was added to the alginate matrix. Furthermore, the presence of SCs also enhanced regeneration and there was an additive effect when both SCs and fibronectin were combined with alginate. In conclusion, the addition of fibronectin to alginate hydrogel matrix contributed to improve nerve regeneration, supporting SC viability and augmenting their effect on axonal growth when transplanted in a bioengineered nerve conduit.

Muller, L. J., C. F. Marfurt, et al. (2003). "Corneal nerves: structure, contents and function." Exp Eye Res 76(5): 521-42.
 This review provides a comprehensive analysis of the structure, neurochemical content, and functions of corneal nerves, with special emphasis on human corneal nerves. A revised interpretation of human corneal nerve architecture is presented based on recent observations obtained by in vivo confocal microscopy (IVCM), immunohistochemistry, and ultrastructural analyses of serial-sectioned human corneas. Current data on the neurotransmitter and neuropeptide contents of corneal nerves are discussed, as are the mechanisms by which corneal neurochemicals and associated neurotrophins modulate corneal physiology, homeostasis and wound healing. The results of recent clinical studies of topically applied neuropeptides and neurotrophins to treat neurotrophic keratitis are reviewed. Recommendations for using IVCM to evaluate corneal nerves in health and disease are presented.

Murakami, T., Y. Fujimoto, et al. (2003). "Transplanted neuronal progenitor cells in a peripheral nerve gap promote nerve repair." Brain Res 974(1-2): 17-24.
 A basic experiment of peripheral nerve regeneration using neuronal progenitor cells embedded in collagen gel was performed in a rat sciatic nerve defect. First, when neuronal progenitor cells derived from the fetal rat hippocampus were cultured in atelocollagen-containing medium, neurospheres positive for anti-nestin antibody were confirmed after 8 days. These cells differentiated into astrocytes positive for anti-glial fibrillary acidic protein (GFAP) antibody, oligodendrocytes positive for anti-galactocerebroside (GalC) antibody and neurons positive for anti-neurofilament 200 (NF200) antibody, and they were capable of extending axons. They also differentiated into Schwann-like supportive cells positive for anti-s100 and anti-p75 antibody. Next, a 15-mm defect was prepared in the sciatic nerve of mature rats, and the nerve was bridged with a silicone tube filled with neuronal progenitor cells (1 x 10(5)) embedded in collagen gel. The transplanted neuronal progenitor cells were labeled in advance with 5-bromo-2-deoxyuridine (BrdU). When the regenerated tissue was examined 6 weeks and 10 weeks after grafting, the number and diameter of myelinated fibers were significantly increased compared with a control tube without neuronal progenitor cells. Action potentials were detected in the regenerated nerve. Also, cells positive for both anti-BrdU antibody and anti-S100 or anti-p75 antibody were observed in the regenerated tissue, and part of the grafted neural stem cells were considered to have differentiated into Schwann cell-like supportive cells. From these results neuronal progenitor cells derived from the fetal rat hippocampus are considered to retain their proliferative and differentiating abilities in collagen gel, and when transplanted to a site of peripheral nerve defect, part of them differentiate into supportive cells and they contributed to promotion of axonal regeneration.

Naccari, C. (2003). "Stem cell transplant regenerates neuronal cells in mice." CNS Spectr 8(5): 336-7.
 
Nadareishvili, Z. and J. Hallenbeck (2003). "Neuronal regeneration after stroke." N Engl J Med 348(23): 2355-6.
 
Nagano, T., M. Nakamura, et al. (2003). "Effects of substance P and IGF-1 in corneal epithelial barrier function and wound healing in a rat model of neurotrophic keratopathy." Invest Ophthalmol Vis Sci 44(9): 3810-5.
 PURPOSE: To establish a rat model of neurotrophic keratopathy and to examine the effects of the combination of substance P (SP) and insulin-like growth factor (IGF)-1 on corneal epithelial barrier function and wound healing in this model. METHODS: Corneal denervation was achieved by thermocoagulation of the ophthalmic branch of the trigeminal nerve. A modified Schirmer test was performed without topical anesthesia. Corneal epithelial barrier function was assessed by measurement of fluorescein permeability with an anterior fluorophotometer. Epithelial wound healing was evaluated by measurement of the area of the defect at various times after removal of the entire epithelium. Eye drops containing both 1 mM SP and IGF-1 (1 micro g/mL) were administered six times daily. RESULTS: The Schirmer test result in eyes subjected to trigeminal denervation was lower than that in control eyes. The fluorescein permeability of the corneal epithelium of denervated eyes was increased relative to that of control eyes. Furthermore, trigeminal denervation induced a delay in corneal epithelial wound healing. Application of eye drops containing SP and IGF-1 to denervated corneas restored the fluorescein permeability of the corneal epithelium to control levels and abolished the delay in epithelial wound healing. CONCLUSIONS: A rat model of neurotrophic keratopathy, characterized by reduced tear secretion, loss of corneal sensation, impaired epithelial barrier function, and delayed epithelial wound healing, was established by trigeminal denervation. Treatment with both SP and IGF-1 improved corneal epithelial barrier function and stimulated corneal epithelial wound healing in this model.

Nagao, M., S. Kato, et al. (2003). "Hyperproliferation of synapses on spinal motor neurons of Duchenne muscular dystrophy and myotonic dystrophy patients." Acta Neuropathol (Berl) 106(6): 557-60.
 Synapses on the motor neurons of patients with Duchenne muscular dystrophy (DMD) and myotonic dystrophy (MD) were studied immunohistochemically using antibodies against synaptobrevin and synaptophysin. Some motor neurons showed hyperproliferation of synapses on the soma and the proximal dendrites. Hyperproliferated synapses were non-cholinergic, because they were not identified with antibodies against vesicular acetylcholine transporter. Regeneration of motor nerve terminals in patients with muscular diseases may affect reorganization of the subsets of central synapses.

Nakamura, M., M. Kawahara, et al. (2003). "Restoration of corneal epithelial barrier function and wound healing by substance P and IGF-1 in rats with capsaicin-induced neurotrophic keratopathy." Invest Ophthalmol Vis Sci 44(7): 2937-40.
 PURPOSE: To investigate the effects of topical application of the combination of substance P (SP) and insulin-like growth factor (IGF)-1 on corneal epithelial barrier function and epithelial wound closure in rats with capsaicin-induced neurotrophic keratopathy. METHODS: Neonatal rats were injected subcutaneously with a single dose of capsaicin to induce neurotrophic keratopathy. Corneal epithelial barrier function was evaluated with an anterior fluorophotometer. Tear fluid secretion was measured by the Schirmer test. Corneal epithelial wound healing was determined by measurement of the size of the epithelial defect after debridement of the entire epithelium. The combination of SP (1 mM) and IGF-1 (1 micro g/mL) in phosphate-buffered saline was administered in eye drops six times daily. RESULTS: Corneal epithelial barrier function was impaired and corneal epithelial wound healing was delayed in rats injected with capsaicin. The application of eye drops containing the combination of SP and IGF-1 to capsaicin-injected rats resulted in a significant improvement in corneal epithelial barrier function compared with that apparent in capsaicin-injected animals that received eye drops containing vehicle alone. Such treatment with SP and IGF-1 also significantly increased the rate of corneal epithelial wound closure in capsaicin-injected animals. CONCLUSIONS: Topical application of the combination of SP and IGF-1 improved both corneal epithelial barrier function and epithelial wound healing in an animal model of neurotrophic keratopathy.

Nakamura, M. and Y. Toyama (2003). "[Transplantation of neural stem cells into spinal cord after injury]." Nippon Rinsho 61(3): 463-8.
 Recovery from central nervous system damage in adult mammals is hindered by their limited ability to replace lost cells and damaged myelin, and reestablish functional neural connections. However, recent progresses in stem cell biology are making it feasible to induce the regeneration of injured axons after spinal cord injury. Transplantation of in vitro expanded neural stem cells into rat spinal cord 9 days after contusion injury induced their differentiation into neurons and oligodendrocytes, and the functional recovery of skilled forelimb movement. It was partly because the microenvironment within the injured spinal cord at 9 days after injury was more favorable for grafted neural stem cells in terms of their survival and differentiation.

Nasatzky, E., J. Gultchin, et al. (2003). "[The role of surface roughness in promoting osteointegration]." Refuat Hapeh Vehashinayim 20(3): 8-19, 98.
 Scientific evidence that has been gathered in the past 20 years established that certain endosseous dental implants--primarily screw-type implants made of commercially pure titanium can be successfully utilized as anchorage for dental prostheses. In recent years, an effort has been made to simplify the surgical procedure, in order to modify clinical treatment modalities. One of the trends is to increasingly utilize microrough titanium implants. Roughened implant surfaces have a long history in implant dentistry, and the most prominent surface is titanium plasma-sprayed (TPS). In recent years new implant surfaces have emerged, so-called microrough titanium surfaces produced with reducing techniques such as grit-blasting with Al2O3 or TiO2 particles, sandblasting and acid-etching, or acid-etching alone. These different titanium surfaces have been tested in numerous in-vivo studies utilizing different animal models. Summarizing the results of these studies, it can be concluded that there is currently sufficient evidence that titanium implants with a microrough surfaces achieve a faster bone integration, a higher percentage of Bone implant Contact (BIC), and a higher resistance to shear documented with higher Removal Torque Values (RTV) when compared with titanium implants with a polished or machined surface. In order to understand the mechanism through which surface roughness modulates its effects mentioned above, recent studies used in-vitro experimental methods to study cell response to implant surface topography. These studies have shown that osteoblasts are sensitive to surface roughness, exhibiting decreased proliferation and a more differentiated phenotype on rougher surfaces. PGE2 production is enhanced on rough surfaces, as is the production of TGF beta 1, suggesting that surface roughness can mediate autocrine and paracrine regulation of osteogenesis. Moreover, surface roughness was found to modulate the effect of systemic hormones like 1,25-(OH)2D3 on osteoblasts. The clinical advantages of implants with rough surface were observed in recently conducted clinical trials. It was found, in humans, that roughened titanium implants need shorter healing period before loading, 6-8 (SLA and Osseotite respectively) weeks instead of 12 weeks. The clinical advantages of shorter healing periods are obvious. Moreover, it was found that certain roughened implants can be used in shorter sizes (6-8 mm) then accepted today. The utilization of shorter implants offers the avoidance of extensive surgical procedures such as nerve lateralization in the mandible or sinus grafting in the maxilla. However, sufficient long term documentation is still lacking, and the predictability of such modalities has yet to be examined in long term prospective clinical trials.

Navarro, X., F. J. Rodriguez, et al. (2003). "Engineering an artificial nerve graft for the repair of severe nerve injuries." Med Biol Eng Comput 41(2): 220-6.
 Nerve repair with tubes has a limit to regeneration depending upon the length of the gap. The characteristics of the guide, in terms of permeability, durability and adhesiveness, also influence regeneration. Considering the importance of the cellular component in regeneration, the development of artificial grafts, composed of a biocompatible nerve guide filled with a neurotropic matrix and seeded with Schwann cells (SCs), is an interesting option to enhance nerve regeneration and provide an alternative to the classical autologous nerve graft. We evaluated the ability of SCs transplanted into a nerve guide to improve regeneration after sciatic nerve resection, leaving a 6-mm gap, in the mouse. Syngeneic, isogeneic and autologous SCs were suspended in Matrigel and seeded in resorbable guides, and compared to acellular guides and to nerve autografts. The immunogenicity of the transplanted SCs clearly influenced the outcome. Transplants of autologous SCs resulted in only slightly lower levels of reinnervation than autografts, but higher recovery and number of regenerated axons than transplants of isologous and syngeneic SCs, and than acellular guides. Thus, by combined developments on nerve guides, extracellular matrix components and cell transplantation, an artificial graft has been designed that allows axonal regeneration across long gaps to levels comparable with an autograft.

Neubert, J. K., L. Karai, et al. (2003). "Peripherally induced resiniferatoxin analgesia." Pain 104(1-2): 219-28.
 Selective blockade of nociceptive pathways represents a mechanism-based approach that has attracted a large variety of pharmacological and molecular investigations. A potential site for selective intervention is the primary afferent nociceptive nerve terminal. Binding of resiniferatoxin (RTX) to the vanilloid-1 receptor (VR1) stimulates and then inactivates heat and vanilloid-responsive nerve endings involved in heat and inflammatory pain signaling which can progress to localized degeneration of the peripheral ending followed by regeneration. Application of RTX directly to peripheral nerve endings produces a long term, reversible attenuation of nociceptive transmission. Heat hyperalgesia and mechanical allodynia were assessed prior to injection of RTX into the hindpaw (baseline) and at acute (minutes-hours) and more chronic (days-weeks) times after injection. Acutely, an inverse dose-to-pain response (guarding, licking) for RTX (0.0625-2.0 microg) occurs, followed by selective attenuation of peripheral pain transmission. Thermal nociception was decreased in a concentration-dependent fashion and lasted up to 21 days, without impairing motor function. Administration of RTX blocked both inflammation-induced hyperalgesia and spinal c-Fos induction. The results demonstrate the efficacy and therapeutic potential of reversible, peripheral C-fiber 'inactivation' for intermediate duration pain control.

Ngo, T. T., P. J. Waggoner, et al. (2003). "Poly(L-Lactide) microfilaments enhance peripheral nerve regeneration across extended nerve lesions." J Neurosci Res 72(2): 227-38.
 After injury, axonal regeneration occurs across short gaps in the peripheral nervous system, but regeneration across larger gaps remains a challenge. To improve regeneration across extended nerve defects, we have fabricated novel microfilaments with the capability for drug release to support cellular migration and guide axonal growth across a lesion. In this study, we examine the nerve repair parameters of non-loaded filaments. To examine the influence of packing density on nerve repair, wet-spun poly(L-Lactide) (PLLA) microfilaments were bundled at densities of 3.75, 7.5, 15, and 30% to bridge a 1.0-cm gap lesion in the rat sciatic nerve. After 10 weeks, nerve cable formation increased significantly in the filament bundled groups when compared to empty-tube controls. At lower packing densities, the number of myelinated axons was more than twice that of controls or the highest packing density. In a consecutive experiment, PLLA bundles with lower filament-packing density were examined for nerve repair across 1.4- and 1.8-cm gaps. After 10 weeks, the number of successful regenerated nerves receiving filaments was more than twice that of controls. In addition, nerve cable areas for control groups were significantly less than those observed for filament groups. Axonal growth across 1.4- and 1.8-cm gaps was more consistent for the filament groups than for controls. These initial results demonstrate that PLLA microfilaments enhance nerve repair and regeneration across large nerve defects, even in the absence of drug release. Ongoing studies are examining nerve regeneration using microfilaments designed to release neurotrophins or cyclic AMP.

Nicole, S., B. Desforges, et al. (2003). "Intact satellite cells lead to remarkable protection against Smn gene defect in differentiated skeletal muscle." J Cell Biol 161(3): 571-82.
 Deletion of murine Smn exon 7, the most frequent mutation found in spinal muscular atrophy, has been directed to either both satellite cells, the muscle progenitor cells and fused myotubes, or fused myotubes only. When satellite cells were mutated, mutant mice develop severe myopathic process, progressive motor paralysis, and early death at 1 mo of age (severe mutant). Impaired muscle regeneration of severe mutants correlated with defect of myogenic precursor cells both in vitro and in vivo. In contrast, when satellite cells remained intact, mutant mice develop similar myopathic process but exhibit mild phenotype with median survival of 8 mo and motor performance similar to that of controls (mild mutant). High proportion of regenerating myofibers expressing SMN was observed in mild mutants compensating for progressive loss of mature myofibers within the first 6 mo of age. Then, in spite of normal contractile properties of myofibers, mild mutants develop reduction of muscle force and mass. Progressive decline of muscle regeneration process was no more able to counterbalance muscle degeneration leading to dramatic loss of myofibers. These data indicate that intact satellite cells remarkably improve the survival and motor performance of mutant mice suffering from chronic myopathy, and suggest a limited potential of satellite cells to regenerate skeletal muscle.

Nicolino, S., S. Raimondo, et al. (2003). "Expression of alpha2a-2b neuregulin-1 is associated with early peripheral nerve repair along muscle-enriched tubes." Neuroreport 14(11): 1541-5.
 Using RT-PCR, we have investigated expression of isoforms beta1 (the axonal isoform) and alpha2a-2b (the mesenchymal isoform) of neuregulin-1, one of the most important known trophic factors for Schwann cells, in the rat sciatic nerve repaired by muscle-enriched non-nervous conduits (made by a vein filled with fresh skeletal muscle). Repaired nerves were harvested 2, 6 and 13 days post-operatively. Results showed that while muscle-vein combined grafts were enriched in mRNA coding for alpha2a-2b since the very early regeneration stages, isoform beta1 mRNA was not detectable inside the tubes at day 2 and 6 post-operatively while its expression at day 13 was very slight. These results suggest that Schwann cell survival and activity inside a fresh muscle-enriched non-nervous conduit graft (a key factor for successful nerve regeneration along the graft) may be supported by the mesenchymal isoform of neuregulin-1 during very early repair phases, i.e. when axons are still not present along the tube.

Nie, X., Y. Jin, et al. (2003). "[Expression of CNTFs in facial motoneurons during facial nerve regeneration in rats and their association with TGF-beta and rhBMP-2]." Hua Xi Kou Qiang Yi Xue Za Zhi 21(1): 52-4.
 OBJECTIVE: The aim of this study was to observe the expression of ciliary neurotrophic factors (CNTFs) in the facial motor neurons in rats during facial nerve regeneration. METHODS: The expression amount of CNTFs in eight groups was determined with immunohistochemical staining and image analysis. RESULTS: The expression of CNTFs increased during the process of nerve regeneration, and reached the maximum one-week and one-month after nerve injury respectively. After one month, the intensity of CNTFs reduced gradually. The expression of recombinant human bone morphogenetic protein-2 (rhBMP-2) did not show distinct difference comparing with the control, but beta transforming growth factors (TGF-beta) benefited the expression of CNTFs during nerve regeneration. CONCLUSION: The endogenous CNTFs promotes the axon outgrow during regeneration, and TGF-beta promotes the expression of CNTFs to rescue motoneuron during facial nerve regeneration.

Nieto, A., J. Casas, et al. (2003). "NGF and TGF-beta mRNA expression during pregnancy in a rat corneal wound healing model." Clin Exp Optom 86(4): 239-43.
 BACKGROUND: Growth factors seem to play a major role in corneal wound healing and TGF-beta seems to be associated with abnormal healing after corneal surgical procedures. Few studies have analysed the role of NGF and TGF-beta on corneal wound healing during pregnancy. The aim of the present study was to create an animal model to evaluate the expression of NGF and TGF-betas during corneal wound healing in two groups: control and pregnant rats. METHODS: Corneal mRNA for NGF and the three isoforms of TGF-beta were analysed by RT-PCR, in a time-course experiment on different days after epithelial wounding (2, 7, 14 days) in pregnant and control groups RESULTS: The results show high corneal mRNA expression for NGF and TGF-beta1 without any variation throughout the healing process or pregnancy evolution. However, we detected a different expression of corneal mRNAs for TGF-beta2 and TGF-beta3 in the control group. This data was not detected in the pregnant group. DISCUSSION: Our results suggested that pregnancy could have a relevant role on TGF-beta2 and TGF-beta3 mRNA expression during the corneal wound healing process. Additional research should be performed to corroborate these findings.

Ninkovic, M. and W. Dabernig (2003). "Flap technology for reconstructions of urogenital organs." Curr Opin Urol 13(6): 483-8.
 SUMMARY: PURPOSE OF REVIEW The purpose of this review is to summarize the different reconstructive options for urogenital indications. The development of various flap techniques to restore congenital and acquired urogenital defects is presented.RECENT FINDINGS Various reconstructive techniques have been demonstrated recently. On the basis of the reconstructive requirements, two main techniques can be defined: the standard local or regional flap technique (pedicled flap) and the more sophisticated microvascular free flap technique. Free tissue transplantation (transfer) is a procedure that involves microvascular transplantation of a flap (a fasciocutaneous, muscle or composite flap) in one stage from a donor site in the body to a distant recipient site. The viability of the transplanted flap is maintained by microvascular anastomosis between the flap's vessels (at least one artery and one vein) and recipient vessels. Re-innervation and functioning muscle contraction is achieved by suturing the vessels and a motor nerve in the recipient area to a motor nerve of a free transplanted muscle. After regeneration of the nerve and re-innervation of the transplanted muscle, a functioning free transplanted muscle offers enough contractile capacity and strength to replace the function of the missing muscles at the recipient site. The technique of microvascular free tissue transfer necessitates extensive experience in microvascular technique and this approach could be efficiently applied in cooperation with other specialists. Recent studies show the development and clinical application of these new surgical techniques in urology (e.g. in the treatment of bladder acontractility using innervated free latissimus dorsi muscle and in the use of a free microvascular fillet lower leg flap for the reconstruction of a large pelvic-floor defect).SUMMARY Various reconstructive requirements define the techniques for reconstruction. The main principle is to obtain optimal anatomical and functional reconstruction with minimal donor site morbidity. Depending on the etiology of the defect, different reconstructive options are available to optimize the reconstructive result. Optimal reconstruction might best be achieved by adopting an interdisciplinary approach in which the primary objective is to provide the best possible outcome for each patient. This review presents the main indications for and principles of flap selection according to the reconstructive requirements.

Nithya, M., L. Suguna, et al. (2003). "The effect of nerve growth factor on the early responses during the process of wound healing." Biochim Biophys Acta 1620(1-3): 25-31.
 In this study, we investigated the role of nerve growth factor (NGF)-incorporated collagen on wound healing in rats. Full-thickness excision wounds were made on the back of female rats weighing about 150-160 g. Topical application of NGF-incorporated collagen, at a concentration of 1 microg/1.2 mg collagen/cm(2), once a day, for 10 days resulted in complete healing of wounds on the 15th day. The concentrations of collagen, hexosamine and uronic acid in the granulation tissue were determined. The NGF-incorporated collagen-treated rats required shorter duration for the healing with an increased rate of wound contraction. Histological and electron microscopical evaluations were also performed, which reveal the activation of fibroblasts and endoplasmic reticulum and therefore increased level of collagen synthesis due to NGF application. These results clearly indicate that the topical application of NGF-incorporated collagen enhanced the rate of healing of excision wounds.

Nodera, H., R. L. Barbano, et al. (2003). "Epidermal reinnervation concomitant with symptomatic improvement in a sensory neuropathy." Muscle Nerve 27(4): 507-9.
 
Nogueira, M. P., D. Paley, et al. (2003). "Nerve lesions associated with limb-lengthening." J Bone Joint Surg Am 85-A(8): 1502-10.
 BACKGROUND: Nerve injury is one of the most serious complications associated with limb-lengthening. We examined the risk, assessment, and treatment of nerve lesions associated with limb-lengthening. METHODS: We retrospectively studied the records on 814 limb-lengthening procedures. Nerve lesions were defined by clinical signs and symptoms of motor function impairment, sensory alterations, referred pain in the distribution of an affected nerve, and/or positive results of quantitative sensory testing with use of a pressure specified sensory device. RESULTS: Seventy-six (9.3%) of the limbs had a nerve lesion. Eighty-four percent of the nerve lesions occurred during gradual distraction, and 16% occurred immediately following surgery. The pressure specified sensory device showed 100% sensitivity and 86% specificity in the detection of nerve injuries. The patients in whom the lesion was diagnosed with this method, or with this method as well as with nerve conduction studies, had significantly faster recovery than did those diagnosed on the basis of clinical symptoms or nerve conduction studies alone (p = 0.02). Patients undergoing double-level tibial lengthening and those with skeletal dysplasia were at higher risk for nerve lesions (77% and 48%, respectively). Nerve decompression was performed in fifty-three cases (70%). The time between the diagnosis and the surgical decompression was strongly associated with the time to recovery (p = 0.0005). Complete clinical recovery was achieved in seventy-four of the seventy-six cases. CONCLUSIONS: Early detection based on signs and symptoms or testing with a pressure specified sensory device improves the prognosis for nerve injury that occurs during limb-lengthening. Of the methods that we used to identify neurologic compromise, testing with the pressure specified sensory device was the most sensitive. Aggressive early treatment (slowing the rate of lengthening and/or performing decompression) allows continued lengthening without incurring permanent nerve injury. When indicated, decompression of the affected nerve should be performed as soon as possible, thereby improving the chances of and shortening the time to complete recovery.

O'Brien, D. F., M. Farrell, et al. (2003). "Schwann cell invasion of the conus medullaris: case report." Eur Spine J 12(3): 328-31.
 As Schwann cells possess regenerative capabilities there is intense interest concerning their role in central nervous system (CNS) regeneration. We report on a case of an intramedullary schwannoma involving the conus medullaris and spinal cord above it. We discuss the possible origin of these cells and the mechanisms by which these cells may invade the CNS. We offer imaging and discuss experimental studies to support our hypothesis. This case concerns a 48-year-old man, who presented with a 6-month history of bilateral lower extremity weakness. Magnetic resonance imaging (MRI) revealed an intramedullary tumour extending from the conus to T11. At operation, following laminectomy and durotomy, a schwannoma was dissected free from the conus. Total gross resection of tumour was achieved. The patient made an uneventful and full recovery. This case shows that Schwann cells can invade the CNS. Manipulation of the transitional zone astrocytic barrier may offer a potential avenue for Schwann cells to enter the CNS in pathological states.

Odaka, M., Y. Uchiyama, et al. (2003). "Evaluation of morphological and functional regeneration of rat nerve-muscle units after temporary and permanent tubulization." Muscle Nerve 28(2): 194-203.
 We compared the ability of temporary and permanent tubing to achieve morphological and functional recovery of nerve-muscle units, following experimental nerve transection (8-mm gap) in rat tibial nerve. Electrical stimulation of the sciatic nerve was used to analyze tension output, evoked electromyogram and conduction-transmission time (CTT) of denervated nerve-muscle units. Morphological analysis of the nerve and muscle was also performed. Within 6 weeks, the nerve gap had been bridged by a thin nerve trunk, and a few myelinated fibers were observed, although there was still no functional recovery. The rats were divided into two groups: permanent tubing (PT) and temporary tubing (TT; tubing subsequently removed). At 10 weeks after the operation, the TT group showed apparently greater thickness of regenerated nerve trunks, significantly higher tension output of plantar flexors, shorter CTT, and heavier muscle mass. These results were consistent with the presence of myelinated fibers in the regenerated nerve trunks, as shown histologically. Thus, removal of the silicone chamber results in faster and better recovery than tubing left permanently in place.

Oi, H., C. Chiba, et al. (2003). "The appearance and maturation of excitatory and inhibitory neurotransmitter sensitivity during retinal regeneration of the adult newt." Neurosci Res 47(1): 117-29.
 Using living slice preparations from newt retinas at different stages of regeneration, we examined the time course of appearance and maturation of neurotransmitter-induced currents with whole-cell patch-clamp methods. Neurons from which currents were recorded were identified by Lucifer Yellow fills. All progenitor cells examined at the regenerating retinas did not express any voltage-gated Na+ currents and responsiveness to excitatory amino acid analogues (AMPA and NMDA) and inhibitory amino acids (GABA and glycine). Voltage-gated Na+ currents were first detected in premature ganglion cells with round cell body located at the most proximal level of the 'intermediate-II' regenerating retina. AMPA- GABA- and glycine-induced currents were simultaneously observed in many premature ganglion cells expressing Na+ channels, but not all, suggesting that the onset of the Na+ channels is slightly earlier than that of excitatory and inhibitory amino acid receptors in regeneration. NMDA-evoked currents were first observed in the 'intermediate-III' regenerating retina just before the synaptogenesis. Pharmacological properties and reversal potential values of the excitatory and inhibitory amino acid responses did not change substantially between regenerating ganglion cells and mature ganglion cells, while rectification properties of current-voltage relations for AMPA and NMDA responses were somewhat different between them.

Oiwa, Y., R. Sanchez-Pernaute, et al. (2003). "Progressive and extensive dopaminergic degeneration induced by convection-enhanced delivery of 6-hydroxydopamine into the rat striatum: a novel rodent model of Parkinson disease." J Neurosurg 98(1): 136-44.
 OBJECT: A striatal dopamine lesion induces progressive nigral degeneration in rodents; however, intrastriatal injection of 6-hydroxydopamine (6-OHDA) causes only limited lesions due to spontaneous regeneration of the neurons that survive. To make an extensive lesion, the authors used a convection-enhanced delivery (CED) method for intrastriatal infusion of 6-OHDA and evaluated the animals for a model of Parkinson disease (PD). METHODS: Different doses of 6-OHDA were infused into the unilateral striatum in rats by using the CED method. The dopaminergic neuronal degeneration was evaluated based on morphological, biochemical, and behavioral measurements until 8 weeks postlesion. Due to the wide distribution of the drug, CED of 20 microg of 6-OHDA into the striatum was sufficient to obtain a progressive and extensive nigrostriatal lesion as defined by morphological (> 80% cell loss in the substantia nigra [SN]) and biochemical (> 95% decrease in striatal dopamine) criteria. The extent of the lesion manifested as a stable turning behavior with amphetamine (> 6 turns/minute) and apomorphine (> 4 turns/minute). It also appeared that at I week postlesion the apoptotic markers were maximal in neurons of the SN. CONCLUSIONS: A rat model of PD with a progressive and extensive dopamine lesion was successfully made by intrastriatal CED of 6-OHDA. In this model, the therapeutic value can be assessed using behavioral, biochemical, and histochemical measurements. The delay of nigral neuronal death with respect to the time of 6-OHDA administration may provide a therapeutic window for testing neuroprotective strategies.

Okada, S. and H. Okano (2003). "[Regulatory mechanisms of neural stem cell and strategies for therapy]." Nippon Rinsho 61(3): 449-56.
 Neural stem cells(NSCs) are multipotential progenitor cells that can generate neurons, astrocytes, and oligodendrocytes, the three major cell types in the central nervous system. Due to their self-renewal activities, NSCs can proliferate in an undifferentiated state in vitro, allowing them to be expanded mitotically and harvested in bulk. Recent advances in stem cell biology have led us to investigate methods for the regenerative manipulation of the damaged CNS. However, there is much that is still not known about regulatory mechanisms of the differentiation and self-renewal of NSCs. In this article, we review some of the basic notions regarding the extracellular factors and signal transduction cascades involved in the differentiation and maintenance of NSCs.

Onteniente, B., S. Rasika, et al. (2003). "Molecular pathways in cerebral ischemia: cues to novel therapeutic strategies." Mol Neurobiol 27(1): 33-72.
 Stroke is one of the leading causes of death and severe disability in most industrialized countries. Despite the extensive research efforts of both academic and industrial laboratories during the last few decades, no changes have been brought about by the design of neuroprotective therapies. The progressive decrease of stroke-induced death and disability is entirely attributable to improvements in the identification and reduction of risk factors. Over the past few years, experimental research has led to the emergence of a wealth of information regarding the complex and interrelated processes of neuronal degeneration and death triggered by ischemia. This unprecedented insight has led to new theories on the mechanisms of ischemic damage, and has suggested new targets and strategies for therapeutic intervention designed to reduce the clinical consequences of stroke. Among current developments, three strategies seem particularly appealing namely, the limitation of initial or secondary neuronal death by inhibition of apoptotic mechanisms, the enhancement of the endogenous capacity of nervous structures to restore lost function, and the replacement of lost cells by transplantation therapy.

Oshitari, T., S. Okada, et al. (2003). "Adenovirus-mediated gene transfer of Bcl-xL impedes neurite regeneration in vitro." Neuroreport 14(12): 1575-8.
 Mouse retinal explants were transfected with recombinant adenovirus vector carrying the green fluorescent protein (GFP) gene and the rat bcl-xL gene (Adeno-Bcl-xL) to determine its ability to protect retinal ganglion cells against apoptotic cell death and to promote retinal ganglion cell neurite regeneration. Adeno-Bcl-xL-incubated retinas had reduced apoptosis compared to controls. However, neurite regeneration in adeno-treated retinas was less than that of vector-free retina. These results suggest that the usefulness of adenovirus vectors for gene therapy for retinal ganglion cells may be limited.

Oshitari, T. and E. Adachi-Usami (2003). "The effect of caspase inhibitors and neurotrophic factors on damaged retinal ganglion cells." Neuroreport 14(2): 289-92.
 To elucidate the role of caspase inhibitors and neurotrophic factors in retinal ganglion cell (RGC) death and regeneration, we cultured mouse retinal explants in the presence of caspase-1, -3, -8, or -9 inhibitors, brain-derived neurotrophic factor (BDNF) and ciliary neurotrophic factor (CNTF) in serum-free culture media. We quantified apoptosis by TUNEL staining in RGCs and assessed the number of regenerating neurites. Apoptosis of RGCs treated with all caspase inhibitors or with neurotrophic factors was significantly reduced and the number of regenerating neurites was significantly greater than controls (p < 0.05). Our findings indicate that caspase-1, -3, -8, -9 play a critical role in explanted RGC death and may be ideal targets of neuroprotection and regeneration of damaged RGCs.

Oshitari, T., S. Okada, et al. (2003). "Adenovirus-mediated gene transfer of Bcl-xL impedes neurite regeneration in vitro." Neuroreport 14(8): 1159-62.
 Mouse retinal explants were transfected with recombinant adenovirus vector carrying the green fluorescent protein (GFP) gene and the rat bcl-x(L) gene (Adeno-Bcl-xL) to determine its ability to protect retinal ganglion cells against apoptotic cell death and to promote retinal ganglion cell neurite regeneration. Adeno-Bcl-xL-incubated retinas had reduced apoptosis compared with controls. However, neurite regeneration in adeno-treated retinas was less than that of vector-free retina. These results suggest that the usefulness of adenovirus vectors for gene therapy for retinal ganglion cells may be limited.

Oster, S. F. and D. W. Sretavan (2003). "Connecting the eye to the brain: the molecular basis of ganglion cell axon guidance." Br J Ophthalmol 87(5): 639-45.
 In the past several years, a great deal has been learnt about the molecular basis through which specific neural pathways in the visual system are established during embryonic development. This review provides a framework for understanding the principles of retinal ganglion cell axon guidance, and introduces some of the families of axon guidance molecules involved. In addition, the potential relevance of retinal axon guidance to human visual developmental disorders, and to retinal axon regeneration, is discussed.

Otteson, D. C. and P. F. Hitchcock (2003). "Stem cells in the teleost retina: persistent neurogenesis and injury-induced regeneration." Vision Res 43(8): 927-36.
 The retina of the adult teleost fish is an important model for studying persistent and injury-induced neurogenesis in the vertebrate central nervous system. All neurons, with the exception of rod photoreceptors, are continually appended to the extant retina from an annulus of progenitors at the margin. Rod photoreceptors, in contrast, are added to differentiated retina only from a lineage of progenitors dedicated to making rods. Further, when the retina is lesioned, the lineage that produces only rods ceases this activity and regenerates retinal neurons of all types. The progenitors that supply neurons at the retinal margin and rod photoreceptors and regenerated neurons in the mature tissue originate from multipotent stem cells. Recent data suggest that the growth-associated neurogenic activity in the retina is regulated as part of the growth hormone/insulin-like growth factor-I axis. This paper reviews recent evidence for the presence of stem cells in the teleost retina and the molecular regulation of neurogenesis and presents a consensus cellular model that describes persistent and injury-induced neurogenesis in the retinas of teleost fish.

Owens, T. (2003). "The enigma of multiple sclerosis: inflammation and neurodegeneration cause heterogeneous dysfunction and damage." Curr Opin Neurol 16(3): 259-65.
 PURPOSE OF REVIEW: The demyelinating disease multiple sclerosis has an autoimmune inflammatory component, which has dominated the description of multiple sclerosis. A degenerative component to multiple sclerosis was always apparent, but was underappreciated until recently. Recent work has brought axonal pathology and brain atrophy into new focus. The purpose of this review is to highlight the relative roles played by the inflammatory and degenerative processes in multiple sclerosis pathology. RECENT FINDINGS: In the past year reports have been published to show that early disability and disease progression correlate with axonal damage, and that brain atrophy resulting from axonal loss is a feature of early multiple sclerosis, and is not restricted to the secondary progressive forms of the disease. Inflammatory mediators (CD8 T cells and antibodies) are implicated in axonal damage, and treatment with steroids or anti-inflammatory therapies reduce brain atrophy, pointing to the involvement of the inflammatory response in the initiation of degeneration. Reduced regenerative capability also contributes to degeneration, and inflammatory responses are shown to inhibit the growth and migration of precursor cells for oligodendrocytes. SUMMARY: Oligodendrocyte precursors are abundant in multiple sclerosis lesions, but fail to remyelinate. Oligodendrocyte growth and regeneration are probably compromised by the action of growth inhibitory signals and lack of growth stimuli. Inflammatory cells and mediators induce axonal loss as well as demyelination. The degenerative response is therefore an integral and early component of multiple sclerosis.

Ozgenel, G. Y. and G. Filiz (2003). "Effects of human amniotic fluid on peripheral nerve scarring and regeneration in rats." J Neurosurg 98(2): 371-7.
 OBJECT: Peripheral nerve repair surgery is still replete with challenges. Despite technical improvements in microsurgery, classic methods of nerve repair have failed to provide satisfactory results. The purpose of this study was to investigate the effects of amniotic fluid from humans on peripheral nerve scarring and regeneration in rats. METHODS: Forty adult Sprague-Dawley rats were used in this study. After the right sciatic nerve in each rat was transected and repaired using an epineural suture procedure, the nerves were divided into two groups according to the solution applied around the repair site: experimental group, 0.3 ml human amniotic fluid (HAF); and control group, 0.3 ml saline. Macroscopic and histological evaluations of peripheral nerve scarring were performed 4 weeks postsurgery. Nerves treated with HAF demonstrated a significant reduction in the amount of scar tissue surrounding the repair site (p < 0.05). No evidence of a reaction against HAF was noted. Functional nerve regeneration was measured once every 2 weeks by using a sciatic function index until 12 weeks postsurgery. Functional recovery in nerves treated with amniotic fluid occurred significantly faster than that in nerves treated with saline (p < 0.05). Peripheral nerve regeneration was evaluated histomorphologically at 12 weeks postsurgery. Nerves treated with amniotic fluid showed significant improvement with respect to the indices of fiber maturation (p < 0.05). CONCLUSIONS: Preliminary data show that HAF enhances peripheral nerve regeneration. The preventive effect of HAF on epineural scarring and the rich content of neurotrophic and neurite-promoting factors possibly contribute to this result.

Pan, W., L. Zhang, et al. (2003). "Selective increase in TNF alpha permeation across the blood-spinal cord barrier after SCI." J Neuroimmunol 134(1-2): 111-7.
 We generated a novel mouse model of spinal cord injury (SCI) by hemisection of the right L1 lumbar spinal cord, measured the permeability of the blood-spinal cord barrier (BSCB), and tested the hypothesis that tumor necrosis factor alpha (TNF alpha) penetrates the injured BSCB by an enhanced transport system. SCI produced stereotypical sensorimotor deficits resembling the classically described Brown-Sequard syndrome. Disruption of the BSCB was reflected by increased spinal cord uptake of radiolabeled albumin from blood; this was transient (immediately after SCI) and confined to the lumbar spinal cord. By contrast, specific increase in the entry of TNF alpha was detected in brain, cervical, thoracic, and lumbar spinal cord at 1 week after SCI, in addition to its immediate and transient increase consistent with barrier disruption. Lack of a second peak of increase in the entry of IL1 beta further supported the specificity of the TNF alpha response. Moreover, enhanced uptake of radiolabeled TNF alpha was suppressed by excess non-radiolabeled TNF alpha, indicating competition of entry via the known transport system for TNF alpha. Therefore, upregulation of the transport system after SCI probably mediates the increased permeation of TNF alpha across the BSCB. Enhanced entry of TNF alpha at 1 week after SCI was concurrent with sensorimotor and gait improvement of the mouse. We conclude that SCI by lumbar hemisection activates the transport system for TNF alpha at the BBB and suggest that selective permeation of TNF alpha may facilitate functional recovery.

Pan, K., M. K. Scott, et al. (2003). "2,3-Diaryl-5-anilino[1,2,4]thiadiazoles as melanocortin MC4 receptor agonists and their effects on feeding behavior in rats." Bioorg Med Chem 11(2): 185-92.
 The melanocortin-4 receptor (MC4) modulates physiological functions such as feeding behavior, nerve regeneration, and drug addiction. Using a high throughput screen based on (125)I-NDP-MSH binding to the human MC4 receptor, we discovered 2,3-diaryl-5-anilino[1,2,4]thiadiazoles 3 as potent and selective MC4 receptor agonists. Through SAR development on the three attached aryl rings, we improved the binding affinity from 174 nM to 4.4 nM IC(50). When delivered intraperitoneally, compounds 3a, 3b, and 3c induced significant inhibition of food intake in a fasting-induced feeding model in rats. When delivered orally, these compounds lost activity, mainly due to rapid metabolism to inactive imidoylthiourea reduction products.

Papalia, I., S. Geuna, et al. (2003). "Morphologic and functional study of rat median nerve repair by terminolateral neurorrhaphy of the ulnar nerve." J Reconstr Microsurg 19(4): 257-64.
 While it has been shown that terminolateral (end-to-side) neurorrhaphy leads to successful functional motor reinnervation of the peripheral territories belonging to the severed nerve, data on the morphology of terminolateral sprouting and on the voluntary control of the motor function restored by terminolateral neurorrhaphy are still partial. In this study, the severed rat median nerve was sutured in an end-to-side fashion to the intact ulnar nerve. The progression of recovery of the flexion of the fingers was assessed by means of the grasping test. Seven months after surgery, the rats were sacrificed, and morphologic and morphometric analysis was performed on the regenerated median nerve and on the donor ulnar nerve. Results of the functional assessment showed that voluntary motor control of the muscles innervated by the median nerve was partially and progressively recovered by terminolateral neurorrhaphy, with a mean strength in the flexion of the fingers that reached about 20 percent of normal before sacrifice. Morphologic and morphometric analysis showed that nerve-fiber regeneration occurred in all repaired median nerves. Signs of nerve fiber atrophy were detected in the ulnar nerve distal to the point of suture, suggesting the possible occurrence of secondary damage to the donor nerve after terminolateral neurorrhaphy that should be taken into consideration in a clinical perspective.

Pardal-Fernandez, J. M., G. Garcia-Alvarez, et al. (2003). "[Peripheral facial paralysis. The value of clinical neurophysiology]." Rev Neurol 36(10): 991-6.
 AIMS: The aim of this study was to review the value of neurophysiological exploration in peripheral facial paralysis in the literature and in our own experience. METHOD: Peripheral facial paralysis is a frequently occurring cranial neuropathy with an acute presentation. Its striking clinical expression is usually in strong contrast to its benign prognosis, except for a small percentage of cases in which the existence of acute injury to the nerve (axonotmesis) will cause it to progress unfavourably, either due to the absence of nerve regeneration or because this is inadequate or insufficient. A neurophysiological study aids diagnosis and the functional and prognostic assessment. CONCLUSIONS: We recommend the utilisation of standardised studies involving a comprehensive electromyographic and electroneurographic evaluation of the facial territory, including reflexology (blink reflex). Use of a suitable methodology in the neurophysiological study of this neuropathy will allow us to complete the diagnosis and to evaluate progress, even from very early days (early surgical approach prior to the onset of nerve degeneration), all of which is decisive in the prognosis of injury throughout the process.

Parulekar, M. V. and J. S. Elston (2003). "Surgery on the nonparetic eye for oculomotor palsy with aberrant regeneration." J Pediatr Ophthalmol Strabismus 40(4): 219-21.
 
Parvizi, J., M. A. Frankle, et al. (2003). "Corrective osteotomy for deformity in Paget disease." J Bone Joint Surg Am 85-A(4): 697-702.
 BACKGROUND: Severe deformity resulting from Paget disease is not uncommon. Malalignment of the extremity may lead to intractable pain, mechanical overload of the neighboring joints, limitation of motion and function, and dysmorphic appearance. Although corrective osteotomy has been used to treat osseous deformities, the outcome of corrective osteotomy for long-bone deformities resulting from Paget disease remains largely unknown. METHODS: The results after twenty-five corrective osteotomies (twenty-two patients), performed between 1975 and 1995, in sixteen tibiae, eight femora, and one radius were evaluated. There were thirteen men and nine women with a mean age of sixty-seven years. The indication for osteotomy was pain in twenty limbs, recurrent stress fractures in three, and limitation of function in two. A variety of osteotomies and fixation methods were used. Two patients underwent simultaneous total hip arthroplasty and proximal femoral osteotomy. RESULTS: Twenty-three of twenty-five osteotomies healed with an average time to union of six months. Both nonunions were in patients who had been managed with intramedullary fixation. The time to union was significantly shorter in metaphyseal osteotomies fixed with plates than in diaphyseal osteotomies (p < 0.04). There was a substantial improvement in the deformities. Satisfaction was rated excellent or good by fourteen patients, fair by six, and poor by two. Complications included a pin-track infection in two patients, peroneal nerve palsy in one, and loss of fixation following external fixation in one. Disease activity, as measured by serum alkaline phosphatase level, and medical treatment with calcitonin and/or bisphosphonates did not have a significant impact on time to union. CONCLUSIONS: Corrective osteotomy for the treatment of severe deformity in Paget disease can be challenging and yet rewarding. A higher prevalence of complications was observed following intramedullary nailing and external fixation. Fracture-healing seems to be particularly protracted in diaphyseal osteotomies.

Passier, R. and C. Mummery (2003). "Origin and use of embryonic and adult stem cells in differentiation and tissue repair." Cardiovasc Res 58(2): 324-35.
 Stem cells are self-renewing, unspecialised cells that can give rise to multiple cell types of all tissues of the body. They can be derived from the embryo, foetus and adult. The ability of stem cells to divide but also to differentiate to specialised cell types like nerve and muscle, have made them candidates on which to base therapies for diseases and disorders for which no, or only partially effective, therapies are available. Replacement of defective or absent cells in defective tissues and organs could represent a cure. Here, we introduce the background to stem cell research and review the present state-of-the-art in stem cell biology, directed differentiation and tissue repair. In particular, we distinguish embryonic versus adult sources of stem cells and data derived from animal versus human experiments in order to place current research and perspectives for clinical application in their correct context.

Patel, S. V. and J. C. Erie (2003). "Aberrant regeneration of corneal nerves after laser in situ keratomileusis." J Cataract Refract Surg 29(2): 387-9.
 We report a case of aberrant regeneration of corneal nerves along the corneal flap interface after myopic laser in situ keratomileusis (LASIK) using confocal microscopy in vivo. The aberrant stromal nerves persisted at the last follow-up, 2 years post LASIK. The short-term clinical outcome was excellent. The long-term clinical effects are unknown.

Pearson, A. G., C. W. Gray, et al. (2003). "ATF3 enhances c-Jun-mediated neurite sprouting." Brain Res Mol Brain Res 120(1): 38-45.
 The AP-1 transcription factor c-Jun is induced in axotomized neurons of the peripheral and central nervous systems, and in both cases upregulation of c-Jun expression has been correlated with axonal regeneration. More recently there has been interest in the c-Jun-related bZIP transcription factor, ATF3, and its function in neurons. ATF3 is also induced in nerve cells in response to axotomy and there is a correlation between increased ATF3 expression and upregulation of c-Jun in surviving neurons. Moreover, c-Jun is able to induce expression of ATF3. We investigated the effect of co-expressing c-Jun and ATF3 in two neuronal-like cell lines to model transcriptional events occurring in axotomized neurons undergoing regeneration. We show that expression of ATF3 with c-Jun significantly enhances c-Jun-mediated neurite sprouting, and that this phenotype is most likely mediated by a physical association of these two transcription factors. Our results suggest that a program of axonal regeneration is initiated when both c-Jun and ATF3 are upregulated in neurons in response to axotomy.

Pecar, D., I. Masic, et al. (2003). "[Rehabilitation of war injuries of the upper extremities with peripheral nerve lesions at the and Praxis Clinic for physical medicine and rehabilitation in Sarajevo]." Med Arh 57(4): 211-3.
 The war injuries are the most frequent multiple, with the difficult distructions of the tissue and the lesions of the peripheral nerves. By the injuries, the lesions of the nerv system represent the delicate problem for the physical medicine and the rehabilitation during the siege of Sarajevo (1992-1995), in the injuries with the lesions of the peripheral nerves, if they are not treated in the frame of the multiple las urgent, they are postponed the operative treatments longer than three months. This is from the aspect of the successfullness, prognostically and therapeutically, the limiting moment. The successfullness of the operative treatment of the lesions of the peripheral nerves significantly depends on that whether the operation was performed incide tree months. The more difficult consequences in these injuries can appear if simultaneously with the taking care of war injuries does not perform adequately surgical treatment adn the in time physical therapy trough the sufficient long time period. On the four year sample was analyzed the success fullness of the rehabilitation of the injuries of the upper extremities with the lesions of the peripheral nerves. Clinically, uniform, we valorized the success fullness of the treatment of all the patients by the marks from 0.5. In the complete sample the excellent success of the rehabilitation we confirmed in more than a half of patients. The better results show the group of the operated patients, in which is performed neuropathia or neurolisis. The capability of the regeneration of the injured peripheral nerves offers the real possibility for extraordinary recovery also in the most difficult injuries, then the preventions of the significant number of the consequences.

Pearson, R. G., Y. Molino, et al. (2003). "Spatial confinement of neurite regrowth from dorsal root ganglia within nonporous microconduits." Tissue Eng 9(2): 201-8.
 Tissue engineering is founded on the concept of controlling the behavior of individual cells to stimulate tissue formation. This control is achieved by mimicking signals that manage natural tissue development or repair. These interdependent signals include cytokine delivery, extracellular matrix interactions, and cell-cell communication. Here, we report on the effect of spatial guidance as a signal for nerve tissue regeneration, using a simple in vitro model. We observe the acceleration of neurite extension from rat dorsal root ganglia within micron-scale tubes. Within these hydrogel-filled conduits, neurites were observed to extend more rapidly than when cultured within the hydrogel alone. The spatial cue also induced a change in tissue architecture, with the cabling of cells within the microconduit. The acceleration of neurite extension was found to be independent of conduit diameter within the range of 200 to 635 microm. Finally, our in vitro model enabled quantification of the effect of combining spatial control and localized nerve growth factor delivery.

Penderis, J., S. A. Shields, et al. (2003). "Impaired remyelination and depletion of oligodendrocyte progenitors does not occur following repeated episodes of focal demyelination in the rat central nervous system." Brain 126(Pt 6): 1382-91.
 It has been hypothesized that the progressive failure of remyelination in chronic multiple sclerosis is, in part, the consequence of repeated episodes of demyelination at the same site, eventually depleting oligodendrocyte progenitor cells (OPCs) and exhausting the remyelinating capacity. We investigated the effect of previous focal, ethidium bromide-induced demyelination of brain stem white matter (with intervening recovery) on the efficiency of the remyelination process during second and third subsequent episodes of demyelination, and the OPC response during a second episode of demyelination. Previous focal demyelinating lesions followed by recovery did not result in any retardation of the remyelination process, nor did they alter the proportion of Schwann cell versus oligodendrocyte remyelination. The OPC response during remyelination was quantified by in situ hybridization using a probe to platelet-derived growth factor-alpha receptor (PDGF alpha R), an OPC-expressed mRNA. Following recovery from focal, toxin-induced CNS demyelination, the OPC density returned to levels equivalent to those in normal white matter. Further more, there was no depletion of OPCs following repeated episodes of focal, toxin-induced CNS demyelination at the same site. These results indicate that repeated CNS demyelination, which has the opportunity to repair in the intervening period, is not characterized by impaired remyelination or depletion of OPCs.

Penkowa, M. and J. Hidalgo (2003). "Treatment with metallothionein prevents demyelination and axonal damage and increases oligodendrocyte precursors and tissue repair during experimental autoimmune encephalomyelitis." J Neurosci Res 72(5): 574-86.
 Experimental autoimmune encephalomyelitis (EAE) is an animal model for the human demyelinating disease multiple sclerosis (MS). EAE and MS are characterized by significant inflammation, demyelination, neuroglial damage, and cell death. Metallothionein-I and -II (MT-I + II) are antiinflammatory and neuroprotective proteins that are expressed during EAE and MS. We have shown recently that exogenous administration of Zn-MT-II to Lewis rats with EAE significantly reduced clinical symptoms and the inflammatory response, oxidative stress, and apoptosis of the infiltrated central nervous system areas. We show for the first time that Zn-MT-II treatment during EAE significantly prevents demyelination and axonal damage and transection, and stimulates oligodendroglial regeneration from precursor cells, as well as the expression of the growth factors basic fibroblast growth factor (bFGF), transforming growth factor (TGF)beta, neurotrophin-3 (NT-3), NT-4/5, and nerve growth factor (NGF). These beneficial effects of Zn-MT-II treatment could not be attributable to its zinc content per se. The present results support further the use of Zn-MT-II as a safe and successful therapy for multiple sclerosis.

Penkowa, M., J. Camats, et al. (2003). "Metallothionein-I overexpression alters brain inflammation and stimulates brain repair in transgenic mice with astrocyte-targeted interleukin-6 expression." Glia 42(3): 287-306.
 Transgenic expression of IL-6 in the CNS under the control of the GFAP gene promoter, glial fibrillary acidic protein-interleukin-6 (GFAP-IL-6) mice, raises an inflammatory response and causes significant brain damage. However, the results obtained in the GFAP-IL-6 mice after a traumatic brain injury, such as a cryolesion, demonstrate a neuroprotective role of IL-6. Thus, the GFAP-IL-6 mice showed faster tissue repair and decreased oxidative stress and apoptosis compared with control litter-mate mice. The neuroprotective factors metallothionein-I+II (MT-I+II) were upregulated by the cryolesion to a higher extent in the GFAP-IL-6 mice, suggesting that they could be related to the neuroprotection afforded by the transgenic expression of IL-6. To examine this possibility, we have crossed GFAP-IL-6 mice with transgenic mice overexpressing MT-I (TgMT), producing double transgenic GFAP-IL-6 TgMT mice. The results obtained after cryolesion in GFAP-IL-6 TgMT mice, as well as in TgMT mice, consistently supported the idea that the increased MT-I+II levels observed in GFAP-IL-6 mice are a fundamental and important mechanism for coping with brain damage. Accordingly, MT-I overexpression regulated the inflammatory response, decreased oxidative stress and apoptosis significantly, and increased brain tissue repair in comparison with either GFAP-IL-6 or control litter-mate mice. Overall, the results demonstrate that brain MT-I+II proteins are fundamental neuroprotective factors.

Penkowa, M., C. Espejo, et al. (2003). "Increased demyelination and axonal damage in metallothionein I+II-deficient mice during experimental autoimmune encephalomyelitis." Cell Mol Life Sci 60(1): 185-97.
 Metallothioneins I+II (MT-I+II) are antioxidant, neuroprotective factors. We previously showed that MT-I+II deficiency during experimental autoimmune encephalomyelitis (EAE) leads to increased disease incidence and clinical symptoms. Moreover, the inflammatory response of macrophages and T cells, oxidative stress, and apoptotic cell death during EAE were increased by MT-I+II deficiency. We now show for the first time that demyelination and axonal damage are significantly increased in MT-I+II deficient mice during EAE. Furthermore, oligodendroglial regeneration, growth cone formation, and tissue repair including expression of trophic factors were significantly reduced in MT-I+II-deficient mice during EAE. Accordingly, MT-I+II have protective and regenerative roles in the brain.

Perez-Gomez, I. and N. Efron (2003). "Change to corneal morphology after refractive surgery (myopic laser in situ keratomileusis) as viewed with a confocal microscope." Optom Vis Sci 80(10): 690-7.
 PURPOSE: This study aimed to look at morphological changes induced by myopic laser in situ keratomileusis (LASIK) in the human cornea using the confocal microscope and to investigate the link between these changes and alterations to corneal sensitivity. METHODS: An in vivo slit-scanning real-time confocal microscope (Tomey ConfoScan P4, Erlangen, Germany) fitted with an Achroplan 40x/0.75 NA immersion objective and a Cochet-Bonnet esthesiometer were used to examine the morphology and sensitivity of the central corneas of six subjects (12 eyes) at an initial visit (before surgery), and at 1 week, 1 month, 3 months, and 6 months after LASIK for myopia. RESULTS: Keratocyte density anterior to the flap interface showed differences between visits (p < 0.0001) and was found to be lower than at the initial visit at 1 week, 1 month, 3 months, and 6 months. Microfolds were noted at the level of the anterior limiting membrane in 11 of 12 eyes after surgery at all visits. Highly reflective flap interface particles were seen in all eyes at all visits after surgery. The subepithelial nerve fiber layer was clearly visible before surgery but could not be imaged in any of the eyes after surgery. Short, unconnected nerve fibers were observed 3 months after surgery; these appeared to form anastomosing interconnections after 6 months. Postsurgical corneal sensitivity was reduced during the first 3 months and recovered to presurgical levels after 6 months. CONCLUSION: LASIK showed a decrease in anterior keratocyte density and microfolds in the anterior limiting membrane, and reflective particles were observed at the flap interface. Corneal sensitivity was depressed during the first 6 months after LASIK surgery; this time course paralleled the appearance of nerve regeneration during this period. Confocal microscopy is capable of providing interesting new insights into the effects of refractive surgery on corneal morphology.

Peters, C. M., S. D. Rogers, et al. (2003). "Endothelin receptor expression in the normal and injured spinal cord: potential involvement in injury-induced ischemia and gliosis." Exp Neurol 180(1): 1-13.
 The endothelins (ETs) are a family of peptides that exert their biological effects via two distinct receptors, the endothelin A receptor (ET(A)R) and the endothelin B receptor (ET(B)R). To more clearly define the potential actions of ETs following spinal cord injury, we used immunohistochemistry and confocal microscopy to examine the protein expression of ET(A)R and ET(B)R in the normal and injured rat spinal cord. In the normal spinal cord, ET(A)R immunoreactivity (IR) is expressed by vascular smooth muscle cells and a subpopulation of primary afferent nerve fibers. ET(B)R-IR is expressed primarily by radial glia, a small population of gray and white matter astrocytes, ependymal cells, vascular endothelial cells, and to a lesser extent in smooth muscle cells. Fourteen days following compression injury to the spinal cord, there was a significant upregulation in both the immunoexpression and number of astrocytes expressing the ET(B)R in both gray and white matter and a near disappearance of ET(B)R-IR in ependymal cells and ET(A)R-IR in primary afferent fibers. Conversely, the vascular expression of ET(A)R and ET(B)R did not appear to change. As spinal cord injury has been shown to induce an immediate increase in plasma ET levels and a sustained increase in tissue ET levels, ETs would be expected to induce an initial marked vasoconstriction via activation of vascular ET(A)R/ET(B)R and then days later a glial hypertrophy via activation of the ET(B)R expressed by astrocytes. Strategies aimed at blocking vascular ET(A)R/ET(B)R and astrocyte ET(B)Rs following spinal cord injury may reduce the resulting ischemia and astrogliosis and in doing so increase neuronal survival, regeneration, and function.

Peterson, S. and E. Bogenmann (2003). "Osmotic swelling induces p75 neurotrophin receptor (p75NTR) expression via nitric oxide." J Biol Chem 278(36): 33943-50.
 Brain injuries by physical trauma, epileptic seizures, or microbial infection upset the osmotic homeostasis resulting in cell swelling (cerebral edema), inflammation, and apoptosis. Expression of the neurotrophin receptor p75NTR is increased in the injured tissue and axon regeneration is repressed by the Nogo receptor using p75NTR as the signal transducer. Hence, p75NTR seems central to the injury response and we wished to determine the signals that regulate its expression. Here, we demonstrate that tonicity mediated cell swelling rapidly activates transcription of the endogenous p75NTR gene and of a p75NTR promoter-reporter gene in various cell types. Transcription activation is independent of de novo protein synthesis and requires the activities of phospholipase C, protein kinase C, and nitric-oxide synthase. Hence, p75NTR is a nitric oxide effector gene regulated by osmotic swelling, thereby providing a strategy for therapeutic intervention to modulate p75NTR functions following injury.

Petitto, J. M., Z. Huang, et al. (2003). "IL-2 gene knockout affects T lymphocyte trafficking and the microglial response to regenerating facial motor neurons." J Neuroimmunol 134(1-2): 95-103.
 Following facial nerve axotomy in mice, T cells cross the intact blood-brain barrier (BBB), home to nerve cell bodies in the facial motor nucleus (FMN), and augment neuroregenerative processes. The pivotal T cell immunoregulatory cytokine, IL-2, appears to have bidirectional effects on neuronal and microglial cell function, suggesting rival hypotheses that IL-2 could either enhance or disrupt processes associated with regeneration of axotomized facial motor neurons. We tested these competing hypotheses by comparing the effect of facial nerve axotomy on C57BL/6-IL-2(-/-) knockout and C57BL/6-IL-2(+/+) wild-type littermates. Since IL-2 may also be produced endogenously in the brain, we also sought to determine whether differences between the knockout and wild-type mice were attributable to loss of IL-2 gene expression in the CNS, loss of peripheral sources of IL-2 and the associated effects on T cell function, or a combination of these factors. To address this question, we bred novel congenic mice with the SCID mutation (mice lacking T cell derived IL-2) that were homozygous for either the IL-2 knockout or wild-type gene alleles (C57BL/6scid-IL-2(-/-) and C57BL/6scid-IL-2(+/+) littermates, respectively). Groups were assessed for differences in (1) T lymphocytes entering the axotomized FMN; (2) perineuronal CD11b(+) microglial phagocytic clusters, a measure of motor neuron death; and (3) activated microglial cells as measured by MHC-II positivity. C57BL/6-IL-2(-/-) knockout mice had significantly higher numbers of T cells and lower numbers of activated MHC-II-positive microglial cells in the regenerating FMN than wild-type littermates, although the number of CD11b(+) phagocytic microglia clusters did not differ. Thus, despite the significant impairment of T cell function known to be associated with loss of peripheral IL-2, the increased number of T cells entering the axotomized FMN appears to have sufficient activity to support neuroregenerative processes. Congenic C57BL/6scid-IL-2(-/-) knockout mice had lower numbers of CD11b(+) microglial phagocytic clusters than congenic C57BL/6scid-IL-2(+/+) wild-type littermates, suggesting that loss of the IL-2 gene in the CNS (and possibly the loss of other unknown sources of the gene) enhanced neuronal regeneration. Further study of IL-2's complex actions in neuronal injury may provide greater understanding of key variables that determine whether or not immunological processes in the brain are proregenerative.

Phillips, R. J., E. A. Baronowsky, et al. (2003). "Long-term regeneration of abdominal vagus: efferents fail while afferents succeed." J Comp Neurol 455(2): 222-37.
 Vagal afferents regenerate, by 18 weeks after subdiaphragmatic transection, to reinnervate the gut and to differentiate into the two types of terminals normally found in the smooth muscle wall of the gastrointestinal (GI) tract (Phillips et al. [2000] J Comp Neurol. 421:325-346). Regeneration, however, is neither complete nor entirely accurate by 18 weeks. Moreover, the capacity of the vagal efferents to reinnervate the GI tract under comparable conditions has not been evaluated. Therefore, to determine whether a more extended postaxotomy survival interval would (1). result in more extensive reinnervation of smooth muscle, (2). facilitate correction of the inaccuracies of the regenerated axons and terminals, and (3). yield motor as well as sensory reinnervation of GI targets, Sprague-Dawley rats received either complete subdiaphragmatic vagotomies (n = 18) or sham surgeries (n = 12). Physiological endpoints that might normalize as vagal elements regenerated, including body weight, daily food intake, size of first daily meal, and metabolic efficiency, were monitored. At 45 weeks after the vagotomies, the animals were randomly assigned to afferent (wheat germ agglutinin-horseradish peroxidase) or efferent (cholera toxin subunit B-horseradish peroxidase) mapping conditions, and labeled axons and terminals in the stomach and first 8 cm of the small intestine were inventoried in whole-mounts. Afferent regeneration was more extensive at 45 weeks than previously observed at 18 weeks after surgery; however, the amount of GI innervation was still not comparable to the intact pattern of the sham rats. Furthermore, abnormal patterns of sensory organization occurred throughout the reinnervated field, with small bundles of axons forming complex tangles and some individual axons terminating in ectopic locations. The presence of growth cone profiles suggested that vagal reorganization was ongoing even 45 weeks after surgery. In contrast to this relatively extensive, albeit incomplete, sensory reinnervation of the gut, motor fibers had failed to reinnervate the GI tract. Thus, dramatic differences exist in the regenerative capacities of the sensory and motor arms of the vagus under the same surgical and maintenance conditions. Furthermore, the functional measures of disordered energy regulation did not normalize over the 45 weeks during which afferent but not efferent innervation was restored.

Pierson, C. R., W. Zhang, et al. (2003). "Proinsulin C-peptide replacement in type 1 diabetic BB/Wor-rats prevents deficits in nerve fiber regeneration." J Neuropathol Exp Neurol 62(7): 765-79.
 We recently reported that early gene responses and expression of cytoskeletal proteins are perturbed in regenerating nerve in type 1 insulinopenic diabetes but not in type 2 hyperinsulinemic diabetes. We hypothesized that these differences were due to impaired insulin action in the former type of diabetes. To test this hypothesis, type 1 diabetic BB/Wor-rats were replaced with proinsulin C-peptide, which enhances insulin signaling without lowering blood glucose. Following sciatic nerve crush injury, early gene responses such as insulin-like growth factor, c-fos, and nerve growth factor were examined longitudinally in sciatic nerve. Neurotrophic factors, their receptors, and beta-tubulin and neurofilament expression were examined in dorsal root ganglia. C-peptide replacement significantly normalized early gene responses in injured sciatic nerve and partially corrected the expression of endogenous neurotrophic factors and their receptors, as well as neuroskeletal protein in dorsal root ganglia. These effects translated into normalization of axonal radial growth and significantly improved axonal elongation of regenerating fibers in C-peptide-replaced BB/Wor-rats. The findings in C-peptide replaced type 1 diabetic rats were similar to those previously reported in hyperinsulinemic and iso-hyperglycemic type 2 BB/Z-rats. We conclude that impaired insulin action may be more important than hyperglycemia in suppressing nerve fiber regeneration in type 1 diabetic neuropathy.

Pierson, C. R., W. Zhang, et al. (2003). "Insulin deficiency rather than hyperglycemia accounts for impaired neurotrophic responses and nerve fiber regeneration in type 1 diabetic neuropathy." J Neuropathol Exp Neurol 62(3): 260-71.
 Diabetic polyneuropathy (DPN) shows more severe functional and structural changes in type 1 than in type 2 human and experimental diabetes. We have previously suggested that these differences may be due to insulin and/or C-peptide deficiencies in type 1 diabetes. To further explore these differences between type I and type 2 DPN, we examined factors underlying nerve fiber regeneration in the hyperinsulinemic type 2 BB/Z-rat and compared these with previous data obtained from the iso-hyperglycemic, insulin and C-peptide-deficient type 1 diabetic BB/Wor-rat. The expression of neurotrophic factors and cytoskeletal proteins were studied in L4 and L5 dorsal root ganglia (DRG) at various time points after sciatic nerve crush. The data were compared to those of nondiabetes-prone BB-rats. Insulin-like growth factor 1 (IGF-1) and TrkA levels were lower in DRG from type 1 than from those of type 2 and control BB-rats. On the other hand, IGF-1 receptor expression was increased at baseline in type 1 BB/Wor-rats and decreased after crush injury, whereas its expression increased after crush injury in both control and type 2 BB/Z-rats. Following crush injury, betaII- and betaIII-tubulins were upregulated in type 2 BB/Z and control rats, which did not occur in type 1 BB/Wor-rats. Furthermore, type 2 BB/Z-rats showed the normal downregulation of low and medium molecular neurofilament (NF-L and NF-M, respectively), which did not occur in type 1 BB/Wor-rats. These findings were associated with significantly milder abnormalities in axonal elongation and caliber growth of regenerating fibers in type 2 compared to type 1 diabetic rats. These data suggest that impaired insulin signaling in type 1 diabetic nerve may be of greater significance in the regulation of neurotrophic and neurocytoskeletal protein synthesis than hyperglycemia in explaining the differences in nerve fiber regeneration between type 2 and type 1 diabetes.

Pietro, G., M. Lepore, et al. (2003). "The influence of mouse tenascin-C on peripheral nerve regeneration and muscle reinnervation through semi-permeable guidance channels." J Hand Surg [Am] 28 Suppl 1: 18.
 
Plant, G. W., C. L. Christensen, et al. (2003). "Delayed transplantation of olfactory ensheathing glia promotes sparing/regeneration of supraspinal axons in the contused adult rat spinal cord." J Neurotrauma 20(1): 1-16.
 The aim of this study was to determine the preferred time and environment for transplantation of olfactory ensheathing glia (OEG) into the moderately contused adult rat thoracic spinal cord. Purified OEG were suspended in culture medium with or without fibrinogen and injected into the contused cord segment at 30 min or 7 days after injury. Control animals received a contusion injury only or injection of only medium 7 days after contusion. The effects on axonal sparing/regeneration and functional recovery were evaluated 8 weeks after injury. The grafts largely filled the lesion site, reducing cavitation, and appeared continuous with the spinal nervous tissue. Whereas in 7d/medium only animals, 54% of spinal tissue within a 2.5-mm-long segment of cord centered at the injury site was spared, significantly more tissue was spared in 0 d/OEG-medium (73%), 0 d/OEG-fibrin (66%), 7 d/OEG-medium (70%), and 7 d/OEG-fibrin (68%) grafted animals. Compared with controls, the grafted animals exhibited more serotonergic axons within the transplant, the surrounding white matter, and the spinal cord up to at least 20 mm caudal to the graft. Retrograde tracing revealed that all but the 0 d/OEG-fibrin graft promoted sparing/regeneration of supraspinal axons compared with controls. Overall, the 7 d/OEG-medium group resulted in the best response, with twice as many labeled neurons in the brain compared with 7 d/medium only controls. Of the labeled neurons, 68% were located in the reticular formation, and 4% in the red, 4% in the raphe, and 5% in the vestibular nuclei. Hindlimb performance was modestly but significantly improved in the 7 d/OEG-medium group. Our results demonstrate that transplantation of OEG into the moderately contused adult rat thoracic spinal cord promotes sparing/regeneration of supraspinal axons and that 7 d transplantation is more effective than acute transplantation of OEG. Our results have relevant implications for future surgical repair strategies of the contused spinal cord.

Platt, C. I., C. A. Krekoski, et al. (2003). "Extracellular matrix and matrix metalloproteinases in sciatic nerve." J Neurosci Res 74(3): 417-29.
 Although matrix metalloproteinases (MMPs) are increasingly being implicated in several pathologies of the nervous system, it is not yet clear what role they play in normal neurobiological processes. We review the expression of extracellular matrix (ECM) components as well as MMPs and tissue inhibitors of metalloproteinases (TIMPs) in the peripheral nervous system. We explore the expression of certain MMPs and the four TIMPs at the mRNA level in the postnatal mouse sciatic nerve. In addition, we have used substrate gel and in situ zymography to determine levels of MMP-2 and -9 and TIMP activity in rat sciatic nerve after crush and during regeneration. A rapid and transient increase in MMP-9 localised at and immediately distal to the site of injury was observed, whereas an increase in MMP-2 activity was delayed, prolonged, and extended proximal and distal to the injury site. This activity coincides with periods of axonal elongation, suggesting that it could act to facilitate axonal extension along the nerve matrix. We also detected multiple species of gelatinolytic inhibitory activity, including TIMP-1 and -3 in control and injured nerve. These activities probably act to prevent uncontrolled gelatinolytic activity, maintaining nerve integrity at the level essential for axonal regrowth.

Pola, R., E. Gaetani, et al. (2003). "Peripheral nerve ischemia: apolipoprotein E deficiency results in impaired functional recovery and reduction of associated intraneural angiogenic response." Exp Neurol 184(1): 264-73.
 Apolipoprotein E-knockout (apoE KO) mice have peripheral sensory nerve defects, reduced and delayed response to noxious thermal stimuli, abnormal morphology of unmyelinated fibers, and impaired blood-nerve and blood-brain barriers. In this study, we show that, compared to wild-type mice, peripheral nerves of apoE KO mice have impaired ability to respond to ischemia, as demonstrated by measurement of motor and sensory conduction velocity. In addition, mice lacking apoE exhibit a deficit of reinnervation of ischemic epidermis, evaluated by immunofluorescent staining for the pan-neuronal marker PGP 9.5. Also regional nerve blood flow, measured by laser Doppler, and intraneural angiogenesis after ischemia are significantly compromised in apoE-deficient mice. Finally, upregulation of the angiogenic cytokine vascular endothelial growth factor (VEGF), which physiologically occurs after ischemia in the peripheral nerve of wild-type mice, is severely impaired in apoE KO mice. Among the several neural defects that have already been described in mice lacking apoE, this is the first demonstration that functional recovery to ischemia is impaired in the peripheral nerves of these animals. This deficit is mirrored by the inability of upregulating VEGF and mounting an appropriate intraneural angiogenic response following injury. These findings provide new evidence of possible interdependent relationships between VEGF, angiogenesis, and nerve function and regeneration and may provide new important information on the role of apoE in the nervous system.

Polydefkis, M., J. W. Griffin, et al. (2003). "New insights into diabetic polyneuropathy." Jama 290(10): 1371-6.
 Patients with complaints of numbness, tingling, and dysesthesias in the toes and feet are frequently referred to neurologists. Often, the only objective evidence for peripheral nerve dysfunction in these patients is limited to small-caliber sensory nerve fibers. On examination these patients may have reduced distal pinprick sensation, and distal leg skin biopsies show loss of small-caliber nerve fibers. Studies focusing on small-caliber nerve fibers have led to a growing impression that neuropathy can be associated with early diabetes or impaired glucose tolerance (IGT). Often, neuropathy can be the presenting symptom of either diabetes or IGT. Furthermore, the oral glucose tolerance test appears to be a more sensitive measure of glucose dysmetabolism in these patients than levels of fasting blood glucose or glycated hemoglobin. Patients with IGT-associated neuropathy may represent an attractive target population for future regenerative studies given that their neuropathy is less severe and presumably more easily reversed than neuropathy occurring in patients with diabetes.Historically, small-caliber fibers have not been extensively evaluated due to a lack of objective measures. Several measures to evaluate these fibers are emerging, including skin biopsy with visualization of epidermal nerve fibers. The accessibility of epidermal nerve fibers makes them an attractive target for nerve injury models, which have potential for development as novel outcome measures. Such approaches may address some of the challenges of past diabetic polyneuropathy trials.

Poulsen, F. R., M. Meyer, et al. (2003). "[Generation of new nerve cells in the adult human brain]." Ugeskr Laeger 165(14): 1443-7.
 Generation of new nerve cells (neurogenesis) is normally considered to be limited to the fetal and early postnatal period. Thus, damaged nerve cells are not expected to be replaced by generation of new cells. The brain is, however, more plastic than previously assumed. This also includes neurogenesis in the adult human brain. In particular two brain regions show continuous division of neural stem and progenitor cells generating neurons and glial cells, namely the subgranular zone of the dentate gyrus and the subventricular zones of the lateral ventricles. From the latter region newly generated neuroblasts (immature nerve cells) migrate toward the olfactory bulb where they differentiate into neurons. In the dentate gyrus the newly generated neurons become functionally integrated in the granule cell layer, where they are believed to be of importance to learning and memory. It is at present not known whether neurogenesis in the adult human brain can be manipulated for specific repair after brain damage.

Previtali, S. C., G. Dina, et al. (2003). "Schwann cells synthesize alpha7beta1 integrin which is dispensable for peripheral nerve development and myelination." Mol Cell Neurosci 23(2): 210-8.
 Defects in laminins or laminin receptors are responsible for various neuromuscular disorders, including peripheral neuropathies. Interactions between Schwann cells and their basal lamina are fundamental to peripheral nerve development and successful myelination. Selected laminins are expressed in the endoneurium, and their receptors are developmentally regulated during peripheral nerve formation. Loss-of-function mutations have confirmed the importance and the role of some of these molecules. Here we show for the first time that another laminin receptor, alpha7beta1 integrin, previously described only in neurons, is also expressed in Schwann cells. The expression of alpha7 appears postnatally, such that alpha7beta1 is the last laminin receptor expressed by differentiating Schwann cells. Genetic inactivation of the alpha7 subunit in mice does not affect peripheral nerve formation or the expression of other laminin receptors. Of note, alpha7beta1 is not necessary for basal lamina formation and myelination. Nonetheless, these data taken together with the previous demonstration of impaired axonal regrowth in alpha7-null mice suggest a possible Schwann cell-autonomous role for alpha7 in nerve regeneration.

Prince, D. J. and R. L. Carlone (2003). "Retinoic acid involvement in the reciprocal neurotrophic interactions between newt spinal cord and limb blastemas in vitro." Brain Res Dev Brain Res 140(1): 67-73.
 The purpose of this study was to investigate the reciprocal neurotrophic interaction between regenerating limb blastemas and spinal cord explants from the newt Notophthalmus viridescens. Axon outgrowth was measured from spinal cord explants in vitro to assess the neurotrophic activity of early to mid-bud stage blastemas after various treatments. When retinoic acid, a vitamin A metabolite, was added to the medium, it increased both the number and length of axons extending from spinal cord explants. Spinal cord explants co-cultured with blastemas that were previously treated with citral, an inhibitor of retinoic acid synthesis, extended significantly fewer axons than control co-cultures. Blastemas, which were denervated by surgical resection of the brachial plexus 48 h before co-culture, also exhibited a significantly weaker neurotrophic activity than did control innervated blastemas. These results are consistent with a reciprocal interaction between blastema mesenchyme and nerves and suggest either a stimulatory or synergistic role for endogenous retinoic acid in the blastema-derived trophic activity.

Pritchard, J., R. A. Hughes, et al. (2003). "Apolipoprotein E genotypes and clinical outcome in Guillain-Barre syndrome." J Neurol Neurosurg Psychiatry 74(7): 971-3.
 BACKGROUND: Polymorphism of the gene encoding the cholesterol transport protein apolipoprotein E (APOE, gene; apoE, protein), known to be involved in axonal regeneration and remyelination, influences outcome after a variety of central nervous system disorders. Apolipoprotein E gene polymorphisms could affect recovery from Guillain-Barre syndrome. OBJECTIVE: To correlate APOE genotypes with residual disability and degree of improvement in Guillain-Barre syndrome, assessed one year after presentation METHODS: 91 patients with the syndrome were recruited from southeast England and their APOE genotypes were determined. RESULTS: There were no clear differences in APOE genotype or allele frequencies when comparing the 91 patients with controls, nor when comparing 81 patients with good outcome and 10 with poor outcome. CONCLUSIONS: APOE genotype did not influence susceptibility to Guillain-Barre syndrome or recovery from it. This may be because our sample size of 91 was not sufficiently large to detect small differences in recovery associated with different APOE genotypes, or because cholesterol transportation is not a crucial rate limiting step in peripheral nerve regeneration.

Privat, A. (2003). "[Repair of the mammalian central nervous system: the "spinal cord" model]." Bull Acad Natl Med 187(2): 345-54; discussion 355-7.
 The central nervous system of adult mammals has been classically considered as structurally rigid, tightly wired, and unable to be repaired. We have shown that there exists a rather considerable degree of intrinsic plasticity due to the neurons themselves, but merely to glial cells and to multipotent stem cells. The spinal cord constitutes a good model on which we could demonstrate, with vascular and traumatic animal paradigms, that an early pharmacologic intervention could reduce significantly the extent of lesions and the subsequent functional deficit. Moreover, we showed that regeneration of severed central axons could occur, provided that the astrocytes' component of the glial scar was modified. Finally, transplants of embryonic neurons were shown to repair the axonal circuitry below a sectioned cord, and to restore reflex functions. All these data point to unprecedented perspectives of efficient therapies in acute and chronic neurological diseases.

Privat, A. (2003). "Astrocytes as support for axonal regeneration in the central nervous system of mammals." Glia 43(1): 91-3.
 Reactive astrocytes are one of the main impediments for axonal regeneration in the central nervous system of mammals. Using mice KO for GFAP and vimentin, we show that reinnervation occurs after an hemisection of the spinal cord, mainly through sprouting of controlateral intact serotoninergic and cortico-spinal axons, thanks to the absence of glial reactivity. This reinnervation is paralleled by the restoration of impaired locomotion of the ipselateral hindleg. Future applications to spinal cord injured patients are discussed.

Puigdellivol-Sanchez, A., A. Prats-Galino, et al. (2003). "Persistence of tracer in the application site--a potential confounding factor in nerve regeneration studies." J Neurosci Methods 127(1): 105-10.
 Selective reinnervation of peripheral targets after nerve injury might be assessed by injecting a first tracer in a target before nerve injury to label the original neuronal population, and applying a second tracer after the regeneration period to label the regenerated population. However, altered uptake of tracer, fading, and cell death may interfere with the results. Furthermore, if the first tracer injected remains in the target tissue, available for "re-uptake" by misdirected regenerating axons, which originally innervated another region, then the identification of the original population would be confused. With the aim of studying this problem, the sciatic nerve of adult rats was sectioned and sutured. After 3 days, to allow the distal axon to degenerate avoiding immediate retrograde transport, one of the dyes: Fast Blue (FB), Fluoro-Gold (FG) or Diamidino Yellow (DY), was injected into the tibial branch of the sciatic nerve, or in the skin of one of the denervated digits. Rats survived 2-3 months. The results showed labelled dorsal root ganglion (DRG) cells and motoneurones, indicating that late re-uptake of a first tracer occurs. This phenomenon must be considered when the model of sequential labelling is used for studying the accuracy of peripheral reinnervation.

Qi, B., Y. Qi, et al. (2003). "Pro-apoptotic ASY/Nogo-B protein associates with ASYIP." J Cell Physiol 196(2): 312-8.
 We have previously shown that ectopic expression of the ASY/Nogo-B gene induced apoptosis in various cancer cell lines. Nogo-A, a splice variant of the ASY, has been reported to have an inhibitory effect on neuronal regeneration in the central nervous system. To investigate the mechanism of ASY-induced apoptosis or inhibition of neuronal regeneration, we cloned a cDNA for the ASY-interacting protein from the human cDNA library using the yeast two-hybrid method, and obtained a cDNA we designated as ASYIP. The ASYIP protein contains two hydrophobic regions and a double lysine endoplasmic reticulum (ER) retrieval motif at its C-terminus, which was shown to be identical to RTN3, a reticulon family protein of unknown function. We showed that ASY and ASYIP proteins formed a complex also in human cells. Mutational analysis indicated that both of the hydrophobic regions of the ASYIP protein were required for the association. By immunofluorescence analysis, the ASYIP protein was shown to be co-localized with ASY in the ER. Characterization of the ASYIP gene may be very useful in clarifying the mechanism of ASY-induced apoptosis or Nogo-involved inhibition of neuronal regeneration in the central nervous system.

Qi, M. L., Y. Wakabayashi, et al. (2003). "Changes in neurocan expression in the distal spinal cord stump following complete cord transection: a comparison between infant and adult rats." Neurosci Res 45(2): 181-8.
 The distal transected cords of infant rats are more permissive for axon extension than those of adults. To elucidate the biomolecular basis for this phenomenon, we examined the expression pattern of neurocan using semi-quantitative reverse transcription polymerase chain reaction and immunostaining in the distal cord of both adult and infant rats after transection. Neurocan is a chondroitin sulfate proteoglycan with well-documented axon growth-inhibitory properties in the central nervous system. Neurocan mRNA was up-regulated in the distal cord of adult rats shortly after transection, followed by a longer wide distribution of neurocan immunoreactivity (IR) in both neurons and astrocytes; by contrast, upregulation of neurocan mRNA was not seen in infant rats, although transient expression of neurocan IR was seen in neurons. Combined with the different regenerative capacity of infant and adult rats, the present results suggest that neurocan inhibits spinal cord regeneration.

Qin, Y., S. Nair, et al. (2003). "Gene expression changes in single dentate granule neurons after adrenalectomy of rats." Brain Res Mol Brain Res 111(1-2): 17-23.
 Removal of corticosterone by adrenalectomy induces apoptosis 3 days later, in some, but not all, rat dentate granule cells. We hypothesized that individual dentate cells trigger specific gene expression profiles that partly determine their apoptosis susceptibility. RNA was collected from physiologically characterized granule cells at 2 or 3 days after adrenalectomy or sham operation, and linearly amplified. The amplified RNA was hybridized to cDNA clones of: (1) candidate genes earlier identified after adrenalectomy in whole hippocampi with SAGE; and (2) genes encoding growth factors and their receptors. We observed that based on the entire expression profile, cells relatively resistant to apoptosis 3 days after adrenalectomy clustered together with one-third of cells 2 days after adrenalectomy. Within the group of ADX cells, a limited number of transcript ratios were found to correlate-positively or negatively-with a known risk factor for apoptosis, calcium influx. The overall analysis of physiological properties and multiple gene expression in single cells can narrow down the number of critical genes involved in apoptosis identified with large scale gene screening methods and allows a first impression of their role as being a potential risk factor or neuroprotective.

Quante, M., J. Lorenz, et al. (2003). "Sensory reinnervation of myocutaneous flaps revealed by infrared laser evoked sensations and brain potentials." Neurorehabil Neural Repair 17(1): 58-65.
 The aim of this study was to examine the recovery of sensory function in myocutaneous flaps comparing 2 test methods. Eight flaps in 7 patients were examined by using clinical neurological test procedures (CNT) in comparison with psychophysics and evoked brain potentials (LEP) following infrared laser stimuli. The authors found that only 3 out of 8 flaps in 7 patients exhibited signs of reinnervation when tested with CNT. Three grades of reinnervation appeared in 7 flaps when tested with the laser. Grade 1 indicated the recovery of unmyelinated C-fiber function in 7 flaps accounting for the ability to discriminate laser intensities by different degrees of warmth. Grade 2 appeared in 3 of these flaps and was characterized by the additional ability to sense pinprick pain and the elicitation of late components of LEP mediated by thinly myelinated A delta-nociceptors. Grade 3 involved the additional sensibility for superficial touch indicating the recovery of thickly myelinated A beta-fibers noted in 2 of these flaps. The authors conclude that the LEP method is more sensitive than standard neurological test procedures to objectively document early signs of reinnervation after reconstructive flap surgery. This result is promising to investigate greater patient populations comparing different surgical techniques in future studies.

Rabinovsky, E. D., E. Gelir, et al. (2003). "Targeted expression of IGF-1 transgene to skeletal muscle accelerates muscle and motor neuron regeneration." Faseb J 17(1): 53-5.
 Currently, there is no known medical treatment that hastens the repair of damaged nerve and muscle. Using IGF-1 transgenic mice that specifically express human recombinant IGF-1 in skeletal muscle, we test the hypotheses that targeted gene expression of IGF-1 in skeletal muscle enhances motor nerve regeneration after a nerve crush injury. The IGF-1 transgene affects the initiation of the muscle repair process after nerve injury as shown by increased activation of SCA-1positive myogenic stem cells. Increased satellite cell differentiation and proliferation are observed in IGF-1 transgenic mice, shown by increased expression of Cyclin D1, MyoD, and myogenin. Expression of myogenin and nicotinic acetylcholine receptor subunits, initially increased in both wild-type and IGF-1 transgenic mice, are restored to normal levels at a faster rate in IGF-1 transgenic mice, which indicates a rescue of nerve-evoked muscle activity. Expression of the IGF-1 transgene in skeletal muscle results in accelerated recovery of saltatory nerve conduction, increased innervation as detected by neurofilament expression, and faster recovery of muscle mass. These studies demonstrate that local expression of IGF-1 augments the repair of injured nerve and muscle.

Rafiuddin Ahmed, M. and R. Jayakumar (2003). "Peripheral nerve regeneration in RGD peptide incorporated collagen tubes." Brain Res 993(1-2): 208-16.
 This paper describes the regeneration of lesioned sciatic nerve with collagen tubes incorporated with RGD cell-adhesive peptide. Collagen implants of 14 mm were grafted to bridge a gap length of 10 mm nerve defect in a rat model. The regenerated tissues were analyzed histomorphologically. The number of myelinated axons in the regenerated mid-graft of the RGD peptide incorporated groups was statistically significant (p<0.05) than control collagen tube and autograft control after 30 days postoperatively. After 90 days of implantation, the mean counts were still statistically significant in the case of RGD peptide group than control collagen and autograft groups. Immunofluorescence studies demonstrated the staining of S100 proteins in the peripherally located cells indicating the proliferation of Schwann cells in the early days of regeneration. The staining pattern of integrin-alphaV was observed mostly in the perineurial regions in close proximity to the RGD peptide incorporated collagen tubes. Other studies like sciatic functional index, conduction velocity at 90 days postoperatively suggest complete regeneration of lesioned nerves with RGD incorporated collagen implants.

Raisman, G. (2003). "A promising therapeutic approach to spinal cord repair." J R Soc Med 96(6): 259-61.
 
Rajan, B., M. Polydefkis, et al. (2003). "Epidermal reinnervation after intracutaneous axotomy in man." J Comp Neurol 457(1): 24-36.
 Two distinct patterns of reinnervation occur after injury to the cutaneous nerves: regenerative growth of the injured nerve and "collateral sprouting" of neighboring intact nerves. We describe two complementary models of regrowth of transected small sensory fibers in human skin. The "incision" model uses a circular incision that transects the subepidermal plexus, resulting in Wallerian degeneration of the nerve fibers that enter the incised cylinder, leaving a defined zone of denervated dermis and epidermis. The "excision" model utilizes an identical incision, followed by removal of the incised cylinder of skin, leaving a denervated area in which Schwann cells are absent. In the incision model, the earliest reinervation of denervated epidermis occurred by collateral sprouting from the terminals of epidermal axons from just outside the incision line. These axon terminals extended horizontally across the incision line and through the superficial layers of the epidermis, beneath the stratum corneum. By 13 days, numerous regenerating axons appeared in the deeper dermis derived from transected axons. These regenerating axons grew toward and ultimately into the epidermis, so that epidermal axonal density had normalized by 30-75 days. The invasion of these axons was associated with regression of the horizontally growing collateral sprouts. In the excision model, new fibers arose by terminal elongation of the epidermal axons outside the incision line, as in the incision model, and especially by collateral branching of epidermal fibers at the incision margins. These collaterals reached the epidermal surface of the basal lamina at the dermal-epidermal junction and then grew slowly toward the center of the denervated circle. In contrast to the incision model, however, complete reinnervation was not achieved even after 23 months. These models can be used to study reinnervation of denervated skin in man in different injury models and have relevance for exploring the stimuli for axonal growth and remodeling.

Ramer, M. S., E. J. Bradbury, et al. (2003). "Glial cell line-derived neurotrophic factor increases calcitonin gene-related peptide immunoreactivity in sensory and motoneurons in vivo." Eur J Neurosci 18(10): 2713-21.
 Calcitonin gene-related peptide (CGRP) is expressed at high levels in roughly 50% of spinal sensory neurons and plays a role in peripheral vasodilation as well as nociceptive signalling in the spinal cord. Spinal motoneurons express low levels of CGRP; motoneuronal CGRP is thought to be involved in end-plate plasticity and to have trophic effects on target muscle cells. As both sensory and motoneurons express receptors for glial cell line-derived neurotrophic factor (GDNF) we sought to determine whether CGRP was regulated by GDNF. Rats were treated intrathecally for 1-3 weeks with recombinant human GDNF or nerve growth factor (NGF) (12 micro g/day) and dorsal root ganglia and spinal cords were stained for CGRP. The GDNF treatment not only increased CGRP immunoreactivity in both sensory and motoneurons but also resulted in hypertrophy of both populations. By combined in situ hybridization and immunohistochemistry we found that, in the dorsal root ganglia, CGRP was up-regulated specifically in neurons expressing GDNF but not NGF receptors following GDNF treatment. Despite the increase in CGRP in GDNF-treated rats, there was no increase in thermal or mechanical pain sensitivity, while NGF-treated animals showed significant decreases in pain thresholds. In motoneurons, GDNF increased the overall intensity of CGRP immunoreactivity but did not increase the number of immunopositive cells. As GDNF has been shown to promote the regeneration of both sensory and motor axons, and as CGRP appears to be involved in motoneuronal plasticity, we reason that at least some of the regenerative effects of GDNF are mediated through CGRP up-regulation.

Ramer, M. S. (2003). "Spontaneous functional viscerosensory regeneration into the adult brainstem." J Neurosci 23(30): 9770-5.
 The dorsal root entry zone of the vagus nerve (vDREZ) is uniquely characterized by peripheral tissue insertions (PTIs) deep to the brainstem surface, consisting of Schwann cells and a reticulum of astrocytic processes. Because Schwann cells permit peripheral axonal regeneration, the capacity of vagal medullary PTIs to allow centripetal regeneration of visceral afferents after vagal dorsal rhizotomy in adult rats was investigated. The present work shows that vagal axons spontaneously regenerate into appropriate and ectopic brainstem nuclei. They accomplish this by first growing along PTIs but then extend along basal laminas of medullary blood vessels. Electrically stimulated regenerated vagal afferents induced Fos expression (indicating functional connectivity) within appropriate but not ectopic nuclei. The unique structure of the vDREZ can thus support spontaneous functional regeneration of visceral primary afferent axons into the adult CNS.

Ramirez, J. J., J. L. Caldwell, et al. (2003). "Adeno-associated virus vector expressing nerve growth factor enhances cholinergic axonal sprouting after cortical injury in rats." J Neurosci 23(7): 2797-803.
 Nerve growth factor (NGF) is known to promote both the survival of cholinergic neurons after injury and the regeneration of damaged cholinergic axons. Recent evidence has implicated NGF in the regulation of cholinergic axonal sprouting by intact neurons projecting to the hippocampus of rats, sustaining a lesion of the entorhinal cortex. We explored the possibility that NGF may regulate this lesion-induced cholinergic sprouting by injecting recombinant adeno-associated virus (rAAV) vector expressing NGF and green fluorescent protein (GFP) into the dentate gyrus of rats that were subsequently given unilateral entorhinal lesions. Sprague Dawley rats were unilaterally injected with (1) rAAV vector expressing NGF and GFP or (2) rAAV vector expressing GFP. Fourteen days after injection, the animals received lesions of the entorhinal area ipsilateral to the virus injection. Four days after lesion, GFP expression and the septodentate sprouting response in the dentate gyrus were assessed. Optical densitometric analyses revealed a significant increase in acetylcholinesterase label (a marker for cholinergic septodentate sprouting) in the ipsilateral outer molecular layer of the dentate gyrus in rats injected with rAAV vector expressing NGF. Thus, NGF-expressing rAAV vector enhanced the sprouting response of intact cholinergic neurons after unilateral entorhinal lesions in rats.

Rammohan, K. W. (2003). "Axonal injury in multiple sclerosis." Curr Neurol Neurosci Rep 3(3): 231-7.
 The pivotal role of axons in the pathophysiology and pathogenesis of multiple sclerosis (MS) is increasingly becoming the focus of our attention. Axonal injury, considered at one time to be a late phenomenon, is now recognized as an early occurrence in the inflammatory lesions of MS. There is converging evidence from histopathologic, as well as magnetic resonance imaging and magnetic resonance spectroscopy studies, that axons play a crucial and dynamic role during the evolution of MS pathology and the development of clinical disability. It has been repeatedly demonstrated that neurologic functional impairment correlates best with axonal, rather than myelin, injury. The pathophysiology of axonal injury remains speculative. Although generally considered to be sequelae of demyelination, it is possible that axonal injury in MS is indeed a primary event. The discovery that axonal injury can be reversible has provided an impetus to institute early therapy. The finding that irreversible axonal transection occurs in early lesions has underscored now, more than ever before, the need to curtail inflammation and the need to institute early treatment with disease-modifying agents. The axon will undoubtedly remain the focus of our attention regarding research on MS now and in the future.

Reinecke, K., R. Lucius, et al. (2003). "Angiotensin II accelerates functional recovery in the rat sciatic nerve in vivo: role of the AT2 receptor and the transcription factor NF-kappaB." Faseb J 17(14): 2094-6.
 The AT2 receptor regulates several functions of nerve cells, e.g., ionic fluxes, cell differentiation, and axonal regeneration, but also modulates programmed cell death. We tested the hypothesis that angiotensin II (ANG II) via its AT2 receptor not only promotes regeneration but also functional recovery after sciatic nerve crush in adult rats. ANG II (10(-7), 10(-9), 10(-11) M) applied locally via osmotic minipumps promoted functional recovery with maximal effects after the lowest concentration. The toe spread distance as a parameter for re-innervation after 20 days was significantly (P<0.01) greater (10.2+/-10.27 mm) compared with the control group (8.73+/-0.16 mm). The response to local electrical stimulation (return of sensorimotor function) was reduced to 14.6 days vs. 17.9 days in the control group (P<0.01). The AT2 receptor antagonist PD 123319 administered alone or in combination with ANG II completely prevented the ANG II-induced recovery, whereas the AT1 receptor antagonist losartan had no effect. Furthermore, ANG II induces, via the AT2 receptor, activation of the transcription factor NF-kappaB in Schwann cells. Histological criteria, morphometric analyses, and electron microscopy confirmed the functional data. These results are the first to present direct evidence for an involvement of the AT2 receptor and NF-kappaB in peripheral nerve regeneration.

Rhodes, K. E., L. D. Moon, et al. (2003). "Inhibiting cell proliferation during formation of the glial scar: effects on axon regeneration in the CNS." Neuroscience 120(1): 41-56.
 Following a CNS lesion many glial cell types proliferate and/or migrate to the lesion site, forming the glial scar. The majority of these cells express chondroitin sulphate proteoglycans (CS-PGs), previously shown to inhibit axonal growth. In this study, in an attempt to diminish glial scar formation and improve axonal regeneration, proliferating cells were eliminated from the lesion site. Adult rats received a continuous infusion of 2% cytosine-D-arabinofuranoside (araC) or saline for 7 days over the lesion site, immediately following a unilateral transection of the right medial forebrain bundle. Additional groups of rats that received subdural infusions prior to the lesion, and lesioned rats which received no infusion, were also compared in the analyses. Animals were killed at 4, 7, 12 or 18 days post-lesion (dpl) and immunohistochemistry was used to determine the effects of these treatments on tyrosine hydroxylase (TH)-lesioned axons, and on the injury response of glial cells. Almost complete elimination of NG2 oligodendrocyte progenitor cells from the lesion site was seen up to 7 dpl in araC-infused animals; reduced numbers of reactive CD11b microglia were also seen but no effects were seen on the injury response of GFAP astrocytes. Significantly more TH axons were seen distal to the lesion in araC-treated brains, but these numbers dwindled by 18 dpl.

Rigoulot, M. A., C. Leroy, et al. (2003). "Prolonged low-dose caffeine exposure protects against hippocampal damage but not against the occurrence of epilepsy in the lithium-pilocarpine model in the rat." Epilepsia 44(4): 529-35.
 PURPOSE: Acute caffeine exposure has proconvulsant effects and worsens epileptic and ischemic neuronal damage. Surprisingly, prolonged caffeine exposure decreases the susceptibility to seizures and the extent of ischemic damage. We explored whether the exposure to a low long-term dose of caffeine could protect the brain from neuronal damage and epileptogenesis in the lithium-pilocarpine model of temporal lobe epilepsy. METHODS: Rats received either plain tap water or water containing caffeine (0.3 g/L) for 15 days before the induction of status epilepticus (SE) by lithium-pilocarpine and for 7 days after SE. The extent of neuronal damage was assessed in the hippocampus and piriform and entorhinal cortices in brain sections stained with thionine and obtained from animals killed 7 days after SE. The latency to spontaneous recurrent seizures was controlled by video monitoring. RESULTS: Caffeine treatment induced a marked, almost total neuroprotection in CA1 and a very limited protection in the hilus of the dentate gyrus, whereas damage in layers III-IV of the piriform cortex was slightly worsened by the treatment. All rats, whether they received caffeine or plain tap water, became epileptic after the same latency (17-19 days). CONCLUSIONS: Thus these data extend the neuroprotective effects of low long-term caffeine exposure to epileptic damage and confirm that the sole protection of the Ammon's horn has no influence on the genesis of spontaneous recurrent seizures in this model.

Robinson, G. A. and R. D. Madison (2003). "Preferential motor reinnervation in the mouse: comparison of femoral nerve repair using a fibrin sealant or suture." Muscle Nerve 28(2): 227-31.
 Previous studies in rat femoral nerve demonstrated that regenerating motor axons preferentially reinnervate a nerve branch to muscle as opposed to skin, a process that has been termed preferential motor reinnervation (PMR). This process has not been previously reported in the mouse, where the use of transgenic animals could be a powerful tool to study the basic mechanisms that determine accuracy of regenerating motor axons. In the mouse, we applied the same nerve repair (suture) and retrograde labeling strategies that successfully demonstrated PMR in the rat femoral nerve but surprisingly were unable to demonstrate PMR. However, if the mouse femoral nerve was repaired with a fibrin sealant, PMR was readily apparent, suggesting that PMR in the mouse is dependent on the method of nerve repair.

Rogers, R. S., T. M. Graziottin, et al. (2003). "Intracavernosal vascular endothelial growth factor (VEGF) injection and adeno-associated virus-mediated VEGF gene therapy prevent and reverse venogenic erectile dysfunction in rats." Int J Impot Res 15(1): 26-37.
 Penile veno-occlusive dysfunction (venogenic erectile dysfunction) is a common cause of erectile dysfunction (ED). We investigated whether vascular endothelial growth factor (VEGF) can be used to prevent and reverse venogenic ED in a rat model. Pharmacological cavernosometry was developed and validated using adult male rats with either arteriogenic or venogenic ED. Castrated animals were treated with intracavernous VEGF as either a recombinant protein (C+VEGF) or adeno-associated virus (AAV)-mediated VEGF gene therapy (C+VEGF gene) in an attempt to prevent the development of venogenic ED. Other animal groups received testosterone replacement (C+testosterone) or intracavernous AAV-LacZ gene (C+LacZ). Animals with documented venogenic ED were treated with intracavernous VEGF in an attempt to reverse their ED. Functional analysis (pharmacological infusion cavernosometry) was performed following treatment. Penile specimens were harvested for immunohistochemistry and electron microscopic evaluation. Castrated rats showed a decrease in papaverine-induced intracavernous pressure and an increase in maintenance and drop rates during pharmacological cavernosometry. These changes were prevented by systemic testosterone and intracavernous VEGF or AAV-VEGF therapy. Moreover, intracavernous VEGF was able to reverse the venogenic ED produced by castration. The quantity of penile smooth muscle detected by alpha actin staining decreased after castration but not in the C+T, C+VEGF, or C+VEGF gene groups. Transmission electron microscopy revealed atrophy of penile smooth muscle cells and nerves in the castrated rats. In VEGF-treated rats, regeneration of smooth muscle and nerves as well as endothelial cell hypertrophy and hyperplasia were the prominent features. In our animal model, systemic testosterone replacement or intracavernous VEGF (protein and VEGF gene) prevented the veno-occlusive dysfunction in castrated animals. In rats with established venous leakage, VEGF treatment reversed the cavernosometric findings of leakage. Intracavernous injection of either VEGF protein or VEGF gene may be a preferred therapy to preserve erectile function in patients in whom testosterone therapy is contraindicated.

Rogers, S. D., C. M. Peters, et al. (2003). "Endothelin B receptors are expressed by astrocytes and regulate astrocyte hypertrophy in the normal and injured CNS." Glia 41(2): 180-90.
 The ability of mammalian central nervous system (CNS) neurons to survive and/or regenerate following injury is influenced by surrounding glial cells. To identify the factors that control glial cell function following CNS injury, we have focused on the endothelin B receptor (ET(B)R), which we show is expressed by the majority of astrocytes that are immunoreactive for glial acid fibrillary protein (GFAP) in both the normal and crushed rabbit optic nerve. Optic nerve crush induces a marked increase in ET(B)R and GFAP immunoreactivity (IR) without inducing a significant increase in the number of GFAP-IR astrocytes, suggesting that the crush-induced astrogliosis is due primarily to astrocyte hypertrophy. To define the role that endothelins play in driving this astrogliosis, artificial cerebrospinal fluid (CSF), ET-1 (an ET(A)R and ET(B)R agonist), or Bosentan (a mixed ET(A)R and ET(B)R antagonist) were infused via osmotic minipumps into noninjured and crushed optic nerves for 14 days. Infusion of ET-1 induced a hypertrophy of ET(B)R/GFAP-IR astrocytes in the normal optic nerve, with no additional hypertrophy in the crushed nerve, whereas infusion of Bosentan induced a significant decrease in the hypertrophy of ET(B)R/GFAP-IR astrocytes in the crushed but not in the normal optic nerve. These data suggest that pharmacological blockade of astrocyte ET(B)R receptors following CNS injury modulates glial scar formation and may provide a more permissive substrate for neuronal survival and regeneration.

Roonprapunt, C., W. Huang, et al. (2003). "Soluble cell adhesion molecule L1-Fc promotes locomotor recovery in rats after spinal cord injury." J Neurotrauma 20(9): 871-82.
 Previous studies suggest that the cell adhesion molecule L1 promotes neurite growth by neutralizing white matter associated inhibitors of axonal growth. We made a soluble chimeric dimer by linking mouse L1 to human Fc. This L1-Fc construct (40 microg/mL) markedly facilitated neurite outgrowth, as well as neuronal adhesion to white matter on frozen sections of spinal cord. We applied L1-Fc intrathecally (200 microg/mL at 0.5 microL/h) to rat spinal cords for 2 weeks after a 25-mm weight drop contusion of the T13 spinal cord. Initial experiments indicated that L1-Fc is present in the spinal cord after 2 weeks of intrathecal infusion and significantly improved locomotor recovery by 6-12 weeks after injury. We then randomized 45 rats to intrathecal infusion of L1-Fc (L1), phosphate-buffered saline controls (PBS), and a mouse monoclonal IgM antibody (M1). By 12 weeks after injury, L1-treated rats recovered significantly (p < 0.005) better locomotor function (BBB score 10.57 +/- 0.25, n = 14) than PBS-treated rats (BBB score 9.00 +/- 0.33, n = 14) or M1-treated (BBB score 8.71 +/- 0.16, n = 14). Only two rats of 22 treated with saline recovered weight-supported ambulation. Of 20 L1-Fc-treated rats, however, 18 recovered weight-supported walking by 12 weeks. The L1-Fc-treated rats also showed more consistent hindlimb contact placing than saline controls. We injected biotinylated dextran amine (BDA) into the motor cortices of 14 rats treated with L1-Fc to label corticospinal axons, comparing these with 13 rats treated with saline. In saline-treated rats, BDA-labeled corticospinal axons often grew up to the impact edge and occasionally into the impact site. L1-treated rats showed longer corticospinal tract growth at the injury site. Three rats had BDA-labeled axons that extended beyond the impact center. One L1-Fc-treated rat showed axonal extension and synapse formation in cord distal to the injury. These results indicate that soluble L1-Fc promotes axonal growth and functional recovery after spinal cord injury. However, the limited corticospinal tract growth across the injury site cannot account for the observed locomotor recovery. Thus, L1 may be stimulating growth of other motor tracts or protecting axons and neurons. More studies are required to elucidate the mechanisms of L1-Fc-induced locomotor recovery.

Rosales-Cortes, M., J. Peregrina-Sandoval, et al. (2003). "Immunological study of a chitosan prosthesis in the sciatic nerve regeneration of the axotomized dog." J Biomater Appl 18(1): 15-23.
 This study demonstrated that when the regeneration of the axotomized sciatic nerve is induced through tubulization with chitosan, this biomaterial does not induce immunostimulation or immunodepression in the dog. Canine females were distributed among three groups: an intact control group which was only isolated, an axotomized control group, and an axotomized group which was tubulized with 3% chitosan prostheses. In vitro culture and phagocytosis tests, as well as IgG and IgM serum concentrations, were determined in peripheral blood on days 0, 15, 30 and 60. The results showed that chitosan implants did not importantly affect the immune response.

Rosales-Cortes, M., J. Peregrina-Sandoval, et al. (2003). "[Regeneration of the axotomised sciatic nerve in dogs using the tubulisation technique with Chitosan biomaterial preloaded with progesterone]." Rev Neurol 36(12): 1137-41.
 INTRODUCTION: Injuries to peripheral nerves can have different causes and may lead to disorders affecting mobility, sensitivity and loss of motor function as they progress. Weiss, in 1944, introduced tubulisation to promote the regeneration of a sectioned nerve. In this study the biomaterial Chitosan was used to induce and stimulate the regeneration of the sciatic nerve in dogs. At the same time, we took advantage of the characteristics offered by Chitosan to include the neurosteroid progesterone in its matrix, as a promoter of axonal growth. AIMS. The aim of our study was to determine the degree of regeneration of the sciatic nerve in dogs when axotomised tubulised with a Chitosan prosthesis steeped in the neurosteroid progesterone. MATERIALS AND METHODS: Young adult female dogs were used to evaluate the regeneration of the sciatic nerve induced at a standard of 15 mm; regeneration was determined by means of an axonal growth chamber. Nerve growth was studied through histological analysis and by electron microscope. RESULTS: The statistical analysis showed that there were no significant differences in the number of myelinated fibres between the experimental groups. The electron microscope images of the transmission in the regenerated nerves in the groups that were tubulised with Chitosan, with and without neurosteroid preloading, revealed an advanced regenerative process. This was evidenced by the fact that collagen fibres, elastin, Schwann cells and both myelinated and non myelinated fibres were observed in all cases. CONCLUSIONS: The regeneration of axotomised, tubulised nerves was achieved regardless of the treatment that was applied. The distal nerve segment that was analysed revealed a similar structure to that of a normal nerve.

Rosen, B. and G. Lundborg (2003). "A new model instrument for outcome after nerve repair." Hand Clin 19(3): 463-70.
 The Model instrument for outcome after nerve repair has so far proved valid and reliable. It supports the authors' hypothesis that the summary of specific limitations of body function agrees with patients' opinions on the impact of the nerve injury on ADL. These features together with its flexibility make the Model instrument for outcome after nerve repair a psychometrically sound and clinically useful diagnosis-specific outcome instrument for routine evaluation after nerve repair. An optimal choice in the future when assessing the outcome after nerve repair may be a combined use of this model and a more generic outcome instrument. Such a protocol would take into account specific body functions and well being in a wider perspective.

Rosen, B. and G. Lundborg (2003). "Early use of artificial sensibility to improve sensory recovery after repair of the median and ulnar nerve." Scand J Plast Reconstr Surg Hand Surg 37(1): 54-7.
 Artificial sensibility based on use of a "tactile glove" which substitutes for lack of sensory afferent inflow with acoustic feedback, was used early after repair of the median and ulnar nerves in a 21-year-old man. After six and 12 months the functional outcome exceeded what is expected in adults, and analysis with calculations for the minimal detectable change (MDC) in tactile gnosis showed a true change. This case highlights the timing of sensory re-education after nerve repair and also emphasises the importance of early restitution of afferent inflow from a denervated hand during rehabilitation.

Rosenstiel, P., P. Schramm, et al. (2003). "Differential effects of immunophilin-ligands (FK506 and V-10,367) on survival and regeneration of rat retinal ganglion cells in vitro and after optic nerve crush in vivo." J Neurotrauma 20(3): 297-307.
 Immunophilins belong to the large family of peptidyl-prolyl-cis-trans-isomerases known to be involved in many cellular processes (e.g., protein trafficking and transcriptional regulation). Beside the widespread therapeutic use of ligands of immunophilins as immunosuppressants, it has been shown that some of these compounds such as FK506 and V-10,367 may mediate neuroprotection and improve axonal regeneration following damage to peripheral nerve fibers. Here, we have analyzed the effects of these two compounds on neurite outgrowth of retinal explants in vitro and on axonal regeneration of retinal ganglion cells, a population of central intrinsic neurons, ten days following optic nerve crush in vivo. FK506 enhanced neurite outgrowth/regrowth in vitro in a dose dependent manner up to 135% (control = 100%), while V-10,367 was more effective (up to 168%). In vivo, intravitreal V-10,367 and FK506 significantly reduced the number of dying retinal ganglion cells as demonstrated by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling. Local application of FK506 into the vitreous body, but not V-10,367, immediately provided after the optic nerve crush induced the elongation of regenerating fibers across the lesion site for around 1.2 mm. Our data provide evidence that the ligands of the FK506-binding proteins FK506 and V-10,367 protect (otherwise dying) retinal ganglion cells from optic nerve crush-induced cell death, promote neurite outgrowth in vitro and that locally applied FK506 enhances the sprouting of axotomized central intrinsic neurons such as retinal ganglion cells in vivo after optic nerve crush.

Rossi, L., P. Deri, et al. (2003). "Expression of DjXnp, a novel member of the SNF2-like ATP-dependent chromatin remodelling genes, in intact and regenerating planarians." Int J Dev Biol 47(4): 293-8.
 SWI/SNF-related complexes include proteins implicated in the regulation of gene expression by chromatin remodelling. We have identified in planarians, invertebrates well-known for their regenerative capability,the cDNA of a novel gene, DjXnp, which encodes a protein of 1,076 amino acids, containing seven helicase domains similar to those found in the SNF2-like family members. Sequence comparison reveals a significant degree of similarity of DjXNP with mammalian XNP/ATRX proteins. In situ hybridization experiments performed on intact and regenerating planarians demonstrated that DjXnp transcripts were distributed in mesenchymal cells and were especially abundant in nerve cells. During anterior regeneration, DjXnp was detected in the blastemal area where the nervous system is newly forming. This expression pattern reveals extensive similarities with that described for mammalian XNP/ATRX, suggesting that these genes may have a conserved function at the cellular level.

Roth, A., R. Gill, et al. (2003). "Temporal and spatial gene expression patterns after experimental stroke in a rat model and characterization of PC4, a potential regulator of transcription." Mol Cell Neurosci 22(3): 353-64.
 We have used the middle cerebral artery occlusion model in the rat in combination with microarray transcript imaging to study changes in gene activity after ischemic stroke. We analyze transcriptional changes in three regions of the affected, ipsilateral brain sphere using contralateral tissues from the same animal as a control over several time points in 180 individual RNA samples. After 1 h transcription factors and signaling molecules are expressed in all tissues followed by the induction of tissue repair-related genes in the cortices which undergo regeneration. Some of these genes are turned on by PC4, which is upregulated in tissues surrounding the infarct core. Interestingly, PC4 is a nerve growth factor (NGF)-inducible gene and has been associated in earlier studies with neuronal growth processes. The expression mode of PC4, the cellular localization of the gene product, and the functional properties of downstream genes induced in vivo and in vitro using transgenic cell lines suggest that PC4 is a regulator of transcription involved in tissue regeneration after ischemic stroke. The novel experimental strategy applied here is suited to provide insight into the molecular mechanisms underlying stroke and tissue regeneration and may enable the discovery of preventive medicines.

Royo, N. C., J. W. Schouten, et al. (2003). "From cell death to neuronal regeneration: building a new brain after traumatic brain injury." J Neuropathol Exp Neurol 62(8): 801-11.
 During the past decade, there has been accumulating evidence of the involvement of passive and active cell death mechanisms in both the clinical setting and in experimental models of traumatic brain injury (TBI). Traditionally, research for a treatment of TBI consists of strategies to prevent cell death using acute pharmacological therapy. However, to date, encouraging experimental work has not been translated into successful clinical trials. The development of cell replacement therapies may offer an alternative or a complementary strategy for the treatment of TBI. Recent experimental studies have identified a variety of candidate cell lines for transplantation into the injured CNS. Additionally, the characterization of the neurogenic potential of specific regions of the adult mammalian brain and the elucidation of the molecular controls underlying regeneration may allow for the development of neuronal replacement therapies that do not require transplantation of exogenous cells. These novel strategies may represent a new opportunity of great interest for delayed intervention in patients with TBI.

Ruiz-Ederra, J., P. F. Hitchcock, et al. (2003). "Two classes of astrocytes in the adult human and pig retina in terms of their expression of high affinity NGF receptor (TrkA)." Neurosci Lett 337(3): 127-30.
 Astrocytes have been implicated in axon guidance and synaptic regeneration in the retina and these processes involve activation of the high affinity nerve growth factor receptor, known as the tyrosine kinase A (TrkA) receptor. The purpose of the present study was to characterize the expression of TrkA in astrocytes of the adult pig and human retina. To this end, sections of human and pig retinas were immunolabeled with a combination of antibodies to glial fibrillary acidic protein (GFAP) and TrkA. Our study revealed that most of the GFAP-positive cells express TrkA, whereas a rare, novel subpopulation of astrocytes was found to be devoid of TrkA. Our results support the idea that astrocytes play an important neurotrophic role in the retina.

Ruitenberg, M. J., G. W. Plant, et al. (2003). "Ex vivo adenoviral vector-mediated neurotrophin gene transfer to olfactory ensheathing glia: effects on rubrospinal tract regeneration, lesion size, and functional recovery after implantation in the injured rat spinal cord." J Neurosci 23(18): 7045-58.
 The present study uniquely combines olfactory ensheathing glia (OEG) implantation with ex vivo adenoviral (AdV) vector-based neurotrophin gene therapy in an attempt to enhance regeneration after cervical spinal cord injury. Primary OEG were transduced with AdV vectors encoding rat brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), or bacterial marker protein beta-galactosidase (LacZ) and subsequently implanted into adult Fischer rats directly after unilateral transection of the dorsolateral funiculus. Implanted animals received a total of 2 x 105 OEG that were subjected to transduction with neurotrophin-encoding AdV vector, AdV-LacZ, or no vector, respectively. At 4 months after injury, lesion volumes were smaller in all OEG implanted rats and significantly reduced in size after implantation of neurotrophin-encoding AdV vector-transduced OEG. All OEG grafts were filled with neurofilament-positive axons, and AdV vector-mediated expression of BDNF by implanted cells significantly enhanced regenerative sprouting of the rubrospinal tract. Behavioral analysis revealed that OEG-implanted rats displayed better locomotion during horizontal rope walking than unimplanted lesioned controls. Recovery of hind limb function was also improved after implantation of OEG that were transduced with a BDNF- or NT-3-encoding AdV vector. Hind limb performance during horizontal rope locomotion did directly correlate with lesion size, suggesting that neuroprotective effects of OEG implants contributed to the level of functional recovery. Thus, our results demonstrate that genetic engineering of OEG not only resulted in a cell that was more effective in promoting axonal outgrowth but could also lead to enhanced recovery after injury, possibly by sparing of spinal tissue.

Russell, C. (2003). "The roles of Hedgehogs and Fibroblast Growth Factors in eye development and retinal cell rescue." Vision Res 43(8): 899-912.
 Knowledge of normal eye development is crucial for the development of retinal rescue strategies. I shall focus on two signalling pathways that affect retinal development. Fibroblast growth factors function in retinal cell proliferation, retinal ganglion cell axon guidance and target recognition, craniofacial patterning and lens induction. Hedgehog proteins are required for progression of the neurogenic wave, cell proliferation, photoreceptor differentiation, retinal ganglion cell axon growth and craniofacial patterning. These signalling pathways have pleiotropic effects, can interact and have the potential to be used therapeutically. The zebrafish model organism may be well suited to studying how signalling pathways interact.

Sahlgren, C. M., A. Mikhailov, et al. (2003). "Cdk5 regulates the organization of Nestin and its association with p35." Mol Cell Biol 23(14): 5090-106.
 The intermediate filament protein nestin is characterized by its specific expression during the development of neuronal and myogenic tissues. We identify nestin as a novel in vivo target for cdk5 and p35 kinase, a critical signaling determinant in development. Two cdk5-specific phosphorylation sites on nestin, Thr-1495 and Thr-316, were established, the latter of which was used as a marker for cdk5-specific phosphorylation in vivo. Ectopic expression of cdk5 and p35 in central nervous system progenitor cells and in myogenic precursor cells induced elevated phosphorylation and reorganization of nestin. The kinetics of nestin expression corresponded to elevated expression and activation of cdk5 during differentiation of myoblast cell cultures and during regeneration of skeletal muscle. In the myoblasts, a disassembly-linked phosphorylation of Thr-316 indicated active phosphorylation of nestin by cdk5. Moreover, cdk5 occurred in physical association with nestin. Inhibition of cdk5 activity-either by transfection with dominant-negative cdk5 or by using a specific cdk5 inhibitor-blocked myoblast differentiation and phosphorylation of nestin at Thr-316, and this inhibition markedly disturbed the organization of nestin. Interestingly, the interaction between p35, the cdk5 activator, and nestin appeared to be regulated by cdk5. In differentiating myoblasts, p35 was not complexed with nestin phosphorylated at Thr-316, and inhibition of cdk5 activity during differentiation induced a marked association of p35 with nestin. These results demonstrate that there is a continuous turnover of cdk5 and p35 activity on a scaffold formed by nestin. This association is likely to affect the organization and operation of both cdk5 and nestin during development.

Sahenk, Z., C. Serrano-Munuera, et al. (2003). "Evidence for impaired axonal regeneration in PMP22 duplication: studies in nerve xenografts." J Peripher Nerv Syst 8(2): 116-27.
 Whether axonal regeneration in Charcot-Marie-Tooth (CMT) neuropathies is impaired has not been addressed in detail. Our studies in nude mice harboring xenografts from patients with different primary Schwann cell (SC) genetic defects suggested an intimate association between the onset of myelination and impairment in the growth capacity of nude mice axons engulfed by the mutant SCs. To assess the effects of peripheral myelin protein 22 (PMP22) gene duplication on the regeneration process, we conducted morphometric studies to generate temporal growth profiles of myelinated axons within the xenografts obtained from CMT1A patients and from healthy controls. Axon size distribution histograms in controls at different time intervals revealed that size differentiation of myelinated fibers within the grafts is established as early as 2 weeks, and that the temporal pattern of myelination of different sized axons has striking similarities to myelination during development. In PMP22 duplication grafts, the onset of myelination is delayed and the regeneration capacity of all fiber sizes is impaired. This defect, however, is most pronounced for the large diameter axons. In addition, significant large fiber loss occurred after 12 weeks with a concomitant new cycle of regeneration of small size axons. These studies show that the PMP22 duplication in SCs have profound effects on the regeneration process, which might be a contributing factor to preferential distal axonal loss.

Saito, I., Y. Oka, et al. (2003). "Promoting nerve regeneration through long gaps using a small nerve tissue graft." Surg Neurol 59(3): 148-54; discussion 154-5.
 BACKGROUND: If nerve tissue is capable of inducing regeneration, as suggested by the neurotropism theory, then even small pieces of nerve tissue should have the potential to induce nerve regeneration. Therefore, long gaps might presumably be bridged via the neurotrophic potential of small pieces of nerve tissue grafted into the middle of the nerve gap. It is necessary to confirm the validity of the neurotropism theory and to also explore the potential usefulness of small nerve grafting through long gaps. METHODS: A small piece of nerve tissue was grafted into a silicone tube bridging a relatively long nerve gap in an attempt to promote nerve regeneration. A 15-mm gap was created in the left sciatic nerve of 31 Wistar rats (8 weeks of age). The experimental groups included one with nonvascularized nerve tissue grafted into a silicone tube with no distal nerve suturing (NV-A), another with vascularized nerve tissue grafted into a silicone tube with no distal nerve suturing (V-A), a third group with nonvascularized nerve tissue grafted into a silicone tube with distal nerve suturing (NV-P), a fourth group with vascularized nerve tissue grafted into a silicone tube with distal nerve suturing (V-P), and a group with no nerve segment grafted into the silicone tube (control). Electrophysiologic and histologic examinations were performed 10 weeks after the operation. RESULTS: No regeneration was obtained in the control group. Nerve regeneration was evident at the proximal end of the tube in the NV-A, V-A, NV-P, and V-P groups, and at the distal end in the NV-P and V-P groups. The degree of distal regeneration was extremely slight in the NV-A and V-A groups. An electrophysiologic examination performed in the NV-P and V-P groups revealed better results in the latter group. CONCLUSION: Small nerve grafts are capable of inducing nerve regeneration even over a long nerve gap, by grafting nerve tissue into the middle of the lesion using a silicone tube.

Sakaue, Y., M. Sanada, et al. (2003). "Amelioration of retarded neurite outgrowth of dorsal root ganglion neurons by overexpression of PKCdelta in diabetic rats." Neuroreport 14(3): 431-6.
 To examine which isoform of protein kinase C (PKC) may be associated with impaired nerve regeneration in diabetes, we compared neurite outgrowth of isolated dorsal root ganglion (DRG) neurons in streptozocin (STZ)-induced diabetic and control rats. Neurite outgrowth was significantly retarded in diabetic neurons. Rottlerin, a PKCdelta specific inhibitor, significantly retracted neurite outgrowth whereas Go6976, an inhibitor specific for classical PKCs, had no effect, suggesting a significant role of PKCdelta in neurite outgrowth of DRG neurons. The expression of phosphorylated PKCdelta, but not total PKCdelta, in DRGs was decreased in diabetic rats. When this reduced expression was restored by overexpressing the PKCdelta in isolated DRG neurons, retardation of neurite outgrowth was significantly reversed in diabetic rats. These results suggest that a decrease in phosphorylated PKCdelta is at least in part responsible for impaired neurite outgrowth in diabetes, and that PKCdelta plays a significant role in the pathogenesis of diabetic neuropathy. This observation provides a useful clue for the treatment of diabetic neuropathy.

Sandoval, M. R. and I. Lebrun (2003). "TsTx toxin isolated from Tityus serrulatus scorpion venom induces spontaneous recurrent seizures and mossy fiber sprouting." Epilepsia 44(7): 904-11.
 PURPOSE: To characterize the long-term behavioral, electroencephalographic (EEG) and histopathologic features after a single TsTx microinjection into the hippocampus of rats. METHODS: TsTx, 2 microg, or 1 microl of 0.1 M phosphate buffer was injected into the right dorsal hippocampus of the rat. EEG records and behavioral observations were made over a period of 10 h after injection. For a period of 4 months, the animals were observed for the occurrence of convulsive seizures. At the end of the experiment, the brains were processed by the neo-Timm and Nissl methods. RESULTS: After intrahippocampal TsTx injection, three distinct phases were observed: (a) an immediate period that lasted 1 day, during which the motor and electrographic seizures characteristic of status epilepticus (SE) were seen; (b) a silent period (31-49 days), characterized by normal EEG and behavior; and (c) a period of spontaneous recurrent seizures (SRSs). The seizure frequency was one to two per week. Four months after TsTx injection, hippocampal neuronal loss and mossy fiber sprouting in the supragranular layer of the dentate gyrus were observed. CONCLUSIONS: The SRSs observed in this study may be associated with the TsTx-induced SE and brain damage. All animals injected with the toxin showed massive pyramidal neuronal loss in the dorsal hippocampus as well as intense gliosis and atrophy. Mossy fiber sprouting in the supragranular layer of the dentate gyrus was observed in those animals that had SRSs. The effects observed may be due, at least in part, to TsTx-enhanced release of glutamate in hippocampal pathways.

Sango, K., A. Oohira, et al. (2003). "Phosphacan and neurocan are repulsive substrata for adhesion and neurite extension of adult rat dorsal root ganglion neurons in vitro." Exp Neurol 182(1): 1-11.
 Phosphacan (PC) and neurocan (NC) are major chondroitin sulfate proteoglycans (CS-PGs) in nervous tissue and are involved in the modulation of cell adhesion and neurite outgrowth during neural development and regeneration. In the present study, we examined the effects of PC and NC on the attachment and neurite extension of adult rat dorsal root ganglion (DRG) neurons in vitro. Treatment with PC and NC on poly-L-lysine (PL) significantly impaired both neuronal attachment and neurite extension in a concentration-dependent manner (10 microg/ml > 1 microg/ml >> 0.1 microg/ml), and they were partially suppressed by chondroitinase ABC (ChABC) digestion. The CS-PGs applied to culture medium (1 microg/ml) also displayed inhibitory effects on neurite extension, which were not altered by ChABC treatment. These results show that PC and NC are repulsive substrata for adhesion and neurite regeneration of adult DRG neurons in vitro and suggest that both chondroitin sulfate moieties and core proteins are responsible for the inhibitory actions of the CS-PGs. We also conducted immunohistochemical analyses with the monoclonal antibodies to core proteins of PC (mAb 6B4) and NC (mAb 1G2), which revealed that only a few neurons in the DRG section were stained with these antibodies. In contrast, most DRG neurons at different stages (12 h, 1 day, 2 days, and 4 days) in culture were immunoreactive to mAb 6B4 and mAb 1G2. Taking these findings together, it is plausible that both CS-PGs expressed in the cultured neurons may play a role in the modulation of attachment, survival, and neurite regeneration.

Sankar, V. and R. Muthusamy (2003). "Role of human amniotic epithelial cell transplantation in spinal cord injury repair research." Neuroscience 118(1): 11-7.
 Human amniotic epithelial cells (HAEC) possess certain properties similar to that of neural and glial cells. In the present work, the potential of HAEC as stem cells for spinal cord injury repair was tested. HAEC obtained from human placenta were labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethyllindocarbocyanine perchlorate (Dil) in the culture medium. These labeled cells were transplanted into the transection cavities in the spinal cord of bonnet monkeys. Results were analyzed after 15 and 60 days of post-transplantation. HAEC cells survived in the monkey spinal cord for up to the maximum period of observation in the present study, i.e. 60 days. HAEC graft was penetrated by the host axons. There was no glial scar at the transection lesion site. Some of the host spinal neurons and axons were labeled with Dil (used to label HAEC) whereas in lesion control group, there was no such host-neuron labeling. This may be either due to the prevention of death in the axotomized neuron's ensuing lesion or due to the neurotrophic effect exhibited by the transplanted HAEC. Further studies would be required to verify these speculations. Therefore from this pilot study it appears that HAEC survive in the transplanted environment, support the growth of host axons through them, prevent the formation of glial scar at the cut ends and may prevent death in axotomized cells or attract the growth of new collateral sprouting. The abovementioned properties, i.e. serving as a suitable milieu for the host axons to grow, preventing glial scar at the lesion site and rescuing axotomized neurons from death were previously reported in the case of neural transplantation studies. Thus it is speculated that HAEC may be having certain properties equal to the beneficial effects of neural tissue in repairing spinal cord injury. Apart from this speculation, there are two more reasons for why HAEC transplantation studies are warranted to understand the long-term effects of such transplantations. First, there was no evidence of immunological rejection probably due to the non-antigenic nature of the HAEC. Second, unlike neural tissue, procurement of HAEC does not involve many legal or ethical problems.

Santo Neto, H., A. J. Martins, et al. (2003). "Axonal sprouting in mdx mice and its relevance to cell and gene mediated therapies for Duchenne muscular dystrophy." Neurosci Lett 343(1): 67-9.
 We investigated whether pre-terminal axons and motor terminals retained their ability to sprout in the murine X-linked muscular dystrophy (mdx). Immunofluorescence confocal microscopy observation of nerve terminals and acetylcholine receptors in mdx muscles with crushed and non-crushed nerves showed that most of the junctions had intraterminal sprouting and that the number of junctions with extraterminal sprouting increased after the nerve crush lesion. Since new dystrophin-positive muscle fibers generated by cell-mediated therapies need to be innervated to proceed with their maturation and dystrophin production, these results suggest that the use of inducing factors to increase the sprouting capacity of nerve terminals could be an additional tool in the success of cell-mediated therapies.

Santos-Benito, F. F. and A. Ramon-Cueto (2003). "Olfactory ensheathing glia transplantation: a therapy to promote repair in the mammalian central nervous system." Anat Rec 271B(1): 77-85.
 A therapy to treat injuries to the central nervous system (CNS) is, to date, a major clinical challenge. The devastating functional consequences they cause in human patients have encouraged many scientists to search, in animal models, for a repair strategy that could, in the future, be applied to humans. However, although several experimental approaches have obtained some degree of success, very few have been translated into clinical trials. Traumatic and demyelinating lesions of the spinal cord have attracted several groups with the same aim: to find a way to promote axonal regeneration, remyelination, and functional recovery, by using a simple, safe, effective, and viable procedure. During the past decade, olfactory ensheathing glia (OEG) transplantation has emerged as a very promising experimental therapy to promote repair of spinal cords, after different types of injuries. Transplants of these cells promoted axonal regeneration and functional recovery after partial and complete spinal cord lesions. Moreover, olfactory ensheathing glia were able to form myelin sheaths around demyelinated axons. In this article, we review these recent advances and discuss to what extent olfactory ensheathing glia transplantation might have a future as a therapy for different spinal cord affections in humans.

Sapieha, P. S., M. Peltier, et al. (2003). "Fibroblast growth factor-2 gene delivery stimulates axon growth by adult retinal ganglion cells after acute optic nerve injury." Mol Cell Neurosci 24(3): 656-72.
 Basic fibroblast growth factor (or FGF-2) has been shown to be a potent stimulator of retinal ganglion cell (RGC) axonal growth during development. Here we investigated if FGF-2 upregulation in adult RGCs promoted axon regrowth in vivo after acute optic nerve injury. Recombinant adeno-associated virus (AAV) was used to deliver the FGF-2 gene to adult RGCs providing a sustained source of this neurotrophic factor. FGF-2 gene transfer led to a 10-fold increase in the number of axons that extended past 0.5 mm from the lesion site compared to control nerves. Detection of AAV-mediated FGF-2 protein in injured RGC axons correlated with growth into the distal optic nerve. The response to FGF-2 upregulation was supported by our finding that FGF receptor-1 (FGFR-1) and heparan sulfate (HS), known to be essential for FGF-2 signaling, were expressed by adult rat RGCs. FGF-2 transgene expression led to only transient protection of injured RGCs. Thus the effect of this neurotrophic factor on axon extension could not be solely attributed to an increase in neuronal survival. Our data indicate that selective upregulation of FGF-2 in adult RGCs stimulates axon regrowth within the optic nerve, an environment that is highly inhibitory for regeneration. These results support the hypothesis that key factors involved in axon outgrowth during neural development may promote regeneration of adult injured neurons.

Scarlato, M., J. Ara, et al. (2003). "Induction of neuropilins-1 and -2 and their ligands, Sema3A, Sema3F, and VEGF, during Wallerian degeneration in the peripheral nervous system." Exp Neurol 183(2): 489-98.
 The neuropilins, NP-1 and NP-2, are coreceptors for Sema3A and Sema3F, respectively, both of which are repulsive axonal guidance molecules. NP-1 and NP-2 are also coreceptors for vascular endothelial growth factor (VEGF). The neuropilins and their ligands are known to play prominent roles in axonal pathfinding, fasciculation, and blood vessel formation during peripheral nervous system (PNS) development. We confirmed a prior report (Exp. Neurol. 172 (2001) 398) that VEGF mRNA levels rise during Wallerian degeneration in the PNS and herein demonstrate that NP-1, NP-2, Sema3A, and Sema3F mRNA levels increase in peripheral nerves distal to a transection or crush injury. In a sciatic nerve crush model, in which axonal regeneration is robust, the highest levels of Sema3F mRNA below the injury site are in the epi- and perineurium. Our results suggest the possibility that the neuropilins and their semaphorin ligands serve to guide, rather than to impede, regenerating axons in the adult PNS.

Scharpf, J., R. Meirer, et al. (2003). "A novel technique for peripheral nerve repair." Laryngoscope 113(1): 95-101.
 OBJECTIVE: To evaluate a novel technique for the repair of neural deficits using a single fascicle to bridge an injury in the rat sciatic nerve. STUDY DESIGN: Twenty-four male Lewis rats were divided into four groups as follows: group 1 (control group), 1.5-cm deficit without repair; group 2, conventional epineural repair with autografts (100% diameter); group 3, nerve repair with large single autograft fascicle (50% diameter); and group 4, nerve repair with small single autograft fascicle (25% diameter). METHODS: Nerve regeneration was evaluated at 3, 6, and 12 weeks by somatosensory evoked potential (SSEP) evaluation and standardized pin-prick and toe-spread tests. Nerve samples were harvested at 12 weeks and stained with toluidine blue to assess the total number of myelinated axons, axon area, and myelin sheath thickness. RESULTS: In group I, the pin-prick and toe-spread tests showed no response at 3, 6, and 12 weeks. Rats in groups 3 and 4 demonstrated significantly better pin-prick test results and a trend toward better toe-spread test responses compared with conventional-repair animals. The SSEP evaluations displayed nondiagnostic waves in rats in group 1 rats. There was no evidence that the other surgery groups differed significantly in median SSEP latencies. Histological evaluation revealed fibrosis in rats in group 1 rats and a significantly higher median number of axons and myelin thickness in the small single fascicle (1296 axons and 4.22 microm, respectively) and large fascicle (2682 axons and 4.62 microm, respectively) groups compared with the conventional autograft group (630 axons and 2.93 microm, respectively). The small fascicle group had a significantly greater mean axon area (58.59 micro m2) than the large fascicle (29.66 micro m2) and conventional autograft (25.35 micro m2) groups. CONCLUSIONS: Peripheral nerve repair using a single fascicle graft resulted in better functional recovery and morphometric outcome without a significant difference in electrophysiological status compared with conventional nerve repair. This technique may provide expanded sources of nerve autografts and alleviate the morbidity of harvesting peripheral nerves from multiple sites for individuals with extensive peripheral nerve injuries.

Schlosshauer, B., E. Muller, et al. (2003). "Rat Schwann cells in bioresorbable nerve guides to promote and accelerate axonal regeneration." Brain Res 963(1-2): 321-6.
 A micro-structured, biodegradable, semipermeable hollow nerve guide implant was developed to bridge nerve lesions. Quantitative comparison of cell migration and axonal growth using time lapse video recording in vitro revealed that axons grow eight times faster than neuritotrophic Schwann cells migrate. To accelerate regeneration, purified Schwann cells are best injected into nerve guides before implantation. Nerve guides made from resorbable poly-lactide-co-glycolide support Schwann cell attachment, cell survival, and axonal outgrowth in vitro. The therapeutic concept aims at the development of an 'intelligent neuroprosthesis' that first mediates regeneration and then disappears.

Schmidt, C. E. and J. B. Leach (2003). "Neural tissue engineering: strategies for repair and regeneration." Annu Rev Biomed Eng 5: 293-347.
 Nerve regeneration is a complex biological phenomenon. In the peripheral nervous system, nerves can regenerate on their own if injuries are small. Larger injuries must be surgically treated, typically with nerve grafts harvested from elsewhere in the body. Spinal cord injury is more complicated, as there are factors in the body that inhibit repair. Unfortunately, a solution to completely repair spinal cord injury has not been found. Thus, bioengineering strategies for the peripheral nervous system are focused on alternatives to the nerve graft, whereas efforts for spinal cord injury are focused on creating a permissive environment for regeneration. Fortunately, recent advances in neuroscience, cell culture, genetic techniques, and biomaterials provide optimism for new treatments for nerve injuries. This article reviews the nervous system physiology, the factors that are critical for nerve repair, and the current approaches that are being explored to aid peripheral nerve regeneration and spinal cord repair.

Schmitt, A. B., S. Breuer, et al. (2003). "Identification of regeneration-associated genes after central and peripheral nerve injury in the adult rat." BMC Neurosci 4(1): 8.
 BACKGROUND: It is well known that neurons of the peripheral nervous system have the capacity to regenerate a severed axon leading to functional recovery, whereas neurons of the central nervous system do not regenerate successfully after injury. The underlying molecular programs initiated by axotomized peripheral and central nervous system neurons are not yet fully understood. RESULTS: To gain insight into the molecular mechanisms underlying the process of regeneration in the nervous system, differential display polymerase chain reaction has been used to identify differentially expressed genes following axotomy of peripheral and central nerve fibers. For this purpose, axotomy induced changes of regenerating facial nucleus neurons, and non-regenerating red nucleus and Clarke's nucleus neurons have been analyzed in an intra-animal side-to-side comparison. One hundred and thirty five gene fragments have been isolated, of which 69 correspond to known genes encoding for a number of different functional classes of proteins such as transcription factors, signaling molecules, homeobox-genes, receptors and proteins involved in metabolism. Sixty gene fragments correspond to genomic mouse sequences without known function. In situ-hybridization has been used to confirm differential expression and to analyze the cellular localization of these gene fragments. Twenty one genes (~15%) have been demonstrated to be differentially expressed. CONCLUSIONS: The detailed analysis of differentially expressed genes in different lesion paradigms provides new insights into the molecular mechanisms underlying the process of regeneration and may lead to the identification of genes which play key roles in functional repair of central nervous tissues.

Schmitt, A. B., S. Breuer, et al. (2003). "Retrograde reactions of Clarke's nucleus neurons after human spinal cord injury." Ann Neurol 54(4): 534-9.
 Successful axon regeneration depends on the expression of regeneration-associated genes by axotomized neurons. Here, we demonstrate, for the first time to our knowledge, the expression of regeneration-associated genes by axotomized human CNS neurons. In situ hybridization and immunohistochemistry showed a transient induction of GAP-43 and c-jun in Clarke's nucleus neurons caudal to traumatic human spinal cord injury. These results support experimental data that nonregenerating central nervous system neurons can temporarily upregulate regeneration-associated genes, reflecting a transient regenerative capacity that fails over time.

Schnaar, R. L. (2003). "Myelin molecules limiting nervous system plasticity." Prog Mol Subcell Biol 32: 125-42.
 
Schuetz, E., K. Rose, et al. (2003). "Regeneration of ganglion cell axons into a peripheral nerve graft alters retinal expression of glial markers and decreases vulnerability to re-axotomy." Restor Neurol Neurosci 21(1-2): 11-8.
 PURPOSE: To compare the effect of cutting the optic nerve versus replacing the cut optic nerve with a peripheral nerve (PN) graft on retinal glial markers, and to determine whether the PN graft can stabilize regenerating retinal ganglion cells (RGCs), thus preventing their death following re-axotomy. METHODS: Retinas harvested after ganglion cell regeneration into a sciatic nerve graft were compared to untreated control retinas and retinas obtained following optic nerve axotomy. Glial-specific proteins such as glial fibrillary acidic protein (GFAP), Bcl-2 and complement-3 receptor (Ox-42) were examined using immunohistochemistry. Ganglion cells that survived the second axotomy were quantified on retinal flat mounts by retrograde labeling from the graft. RESULTS: GFAP expression in astrocytes and Muller cells was elevated in axotomized retinas when compared to controls, and an additional up-regulation in Muller cells was found in retinas following ganglion cell regeneration. Increased GFAP expression in retinas containing regenerated neurons was accompanied by increased Bcl-2 expression with latter being confined to Muller cells. Moreover, re-axotomy of the regenerated axons within the graft did not result in significant retrograde degeneration of RGCs within 28 days. CONCLUSIONS: The data suggest that the graft stabilizes the regenerating RGCs to an extent reminiscent of peripheral neurons, a process that may involve the interaction between neuronal and glial elements.

Schwartz, M. (2003). "Neurodegeneration and neuroprotection in glaucoma: development of a therapeutic neuroprotective vaccine: the Friedenwald lecture." Invest Ophthalmol Vis Sci 44(4): 1407-11.
 
Schweitzer, J., T. Becker, et al. (2003). "Expression of protein zero is increased in lesioned axon pathways in the central nervous system of adult zebrafish." Glia 41(3): 301-17.
 The immunoglobulin superfamily molecule protein zero (P0) is important for myelin formation and may also play a role in adult axon regeneration, since it promotes neurite outgrowth in vitro. Moreover, it is expressed in the regenerating central nervous system (CNS) of fish, but not in the nonregenerating CNS of mammals. We identified a P0 homolog in zebrafish. Cell type-specific expression of P0 begins in the ventromedial hindbrain and the optic chiasm at 3-5 days of development. Later (at 4 weeks) expression has spread throughout the optic system and spinal cord. This is consistent with a role for P0 in CNS myelination during development. In the adult CNS, glial cells constitutively express P0 mRNA. After an optic nerve crush, expression is increased within 2 days in the entire optic pathway. Expression peaks at 1 to 2 months and remains elevated for at least 6 months postlesion. After enucleation, P0 mRNA expression is also upregulated but fails to reach the high levels observed in crush-lesioned animals at 4 weeks postlesion. Spinal cord transection leads to increased expression of P0 mRNA in the spinal cord caudal to the lesion site. The glial upregulation of P0 mRNA expression after a lesion of the adult zebrafish CNS suggests roles for P0 in promoting axon regeneration and remyelination after injury.

Scott, W. A. (2003). "Syncope in a triathlete." Curr Sports Med Rep 2(3): 157-8.
 
Segonds, J. M., J. Y. Alnot, et al. (2003). "[Aseptic non-union of humeral shaft fractures treated by plating and bone grafting]." Rev Chir Orthop Reparatrice Appar Mot 89(2): 107-14.
 PURPOSE OF THE STUDY: Although aseptic non-union of humeral shaft fractures is generally considered to be an exceptional complication, rates in the literature have varied from 1 to 10%. Factors favoring non-union are often related to technical error or inappropriate therapeutic indication. Several types of treatment (orthopedic, locked centromedullary nailing, ascending pinning, plating, external fixation) can be proposed for humeral shaft fractures. In all cases, a precise technique and proper indication are essential for success. We reviewed the cases of 30 patients who underwent surgery for aseptic non-union of humeral shaft fractures between 1995 and 2000. MATERIAL AND METHODS: Mean patient age was 43 years. Oblique and transverse fractures of the middle third of the shaft predominated. All types of treatment had been used but most of the patients had had ascending pinning. All patients were treated with plate fixatin and a cancellous bone graft after identifying the radial nerve. RESULTS: Bone healing was achieved in all patients. Mean delay to healing was 16 weeks with good motion of the shoulder (mean elevation 136 degrees ) and elbow (mean motion 10-130 degrees ). Transient radial paresia recovered spontaneously in two patients. There was one infection. Only two patients complained of a painful arm that was not bothersome for daily activities and did not require long-term analgesia. There were no cases of radial nerve injury. Elbow function improved in 16 patients, was unchanged in 11, and showed limited extension in 3. Shoulder function improved in 15 patients and was unchanged in 15. DISCUSSION: Plate fixation is widely described in the literature for the treatment of humeral non-union. The main complications of this treatment are radial palsy and infection, reported in 5% of the series. Several recent reports have therefore advocated locked nailing or external fixation with an Ilizarov device but these techniques are difficult to use and have their own risks of complications. It is difficult to block rotation and the nail may injure the rotator cuff. Pin tract infection, nerve injury, and prolonged external fixation are other disadvantages. We therefore recommend screw plate fixation with a cancellous bone graft. Our good results combined with the very low rate of complications argue in favor of this therapeutic option.

Sekiya, T., N. Shimamura, et al. (2003). "Effect of topically applied basic fibroblast growth factor on injured cochlear nerve." Neurosurgery 52(4): 900-7; discussion 907.
 OBJECTIVE: Trauma-induced hearing loss after cerebellopontine angle manipulation has been regarded as having a hopeless natural course once it occurs. To challenge such a pessimistic view, we investigated whether pharmacological interventions with basic fibroblast growth factor (bFGF) could ameliorate trauma-induced cochlear nerve degeneration. METHODS: The cerebellopontine angle portion of the cochlear nerve of rats was quantitatively compressed, and bFGF was topically administered for 2 weeks with a bFGF-soaked absorbable sponge and an osmotic minipump. The animals were killed 2 weeks after the compression procedure. The effect of bFGF in ameliorating cochlear neuronal death was evaluated from the residual number of spiral ganglion cells. RESULTS: Cerebellopontine angle cisternal application of bFGF ameliorated cochlear nerve degeneration after the compression. Immunocytochemical studies of FGF receptors indicated that topically administered bFGF was internalized by a receptor-mediated mechanism through FGF receptor-1 and/or FGF receptor-2. CONCLUSION: This report demonstrated that therapeutic application of bFGF was feasible to ameliorate trauma-induced cochlear nerve degeneration. Recent technological advances for deafened ears, such as cochlear implants and auditory brainstem implants, in combination with neurotrophic and/or growth factor therapeutic intervention, would be of great potential benefit for patients with hearing loss.

Selzer, M. E. (2003). "Promotion of axonal regeneration in the injured CNS." Lancet Neurol 2(3): 157-66.
 Molecules that are found in the extracellular environment at a CNS lesion site, or that are associated with myelin, inhibit axon growth. In addition, neuronal changes--such as an age-dependent reduction in concentrations of cyclic AMP--render the neuron less able to respond to axotomy by a rapid, forward, actin-dependent movement. An alternative mechanism, based on the protrusive forces generated by microtubule elongation or the anterograde transport of cytoskeletal elements, may underlie a slower form of axon elongation that happens during regeneration in the mature CNS. Therapeutic approaches that restore the extracellular CNS environment or the neuron's characteristics back to a more embryonic state increase axon regeneration and improve functional recovery after injury. These advances in the understanding of regeneration in the CNS have major implications for neurorehabilitation and for the use of axonal regeneration as a therapeutic approach to disorders of the CNS such as spinal-cord injury.

Serpe, C. J., S. Coers, et al. (2003). "CD4+ T, but not CD8+ or B, lymphocytes mediate facial motoneuron survival after facial nerve transection." Brain Behav Immun 17(5): 393-402.
 The capacity of facial motor neurons (FMN) to survive injury and successfully regenerate is substantially compromised in immunodeficient mice, which lack T and B lymphocytes (). The goal of the present study was to determine which T cell subset (CD4+ and/or CD8+), and whether the B lymphocyte, is involved in FMN survival after nerve injury. All mice were subjected to a right facial nerve axotomy, with the left (uncut) side serving as an internal control. FMN survival, of the right (cut) side, was measured 4 weeks post-operative, and expressed as a percentage of the left (uncut) control side. FMN survival in wild-type mice was 86%+/-1.5. In contrast, FMN survival in CD4 KO mice was 60%+/-2.0. Reconstitution of either CD4 KO mice, or recombinase activating gene-2 knockout (RAG-2 KO) mice (which lack functional T and B cells) with CD4+ T cells alone restored FMN survival to wild-type levels (85%+/-1.2 and 84%+/-2.5, respectively). There was no difference in FMN survival between wild-type, CD8 KO and MmuMT (B cell deficient) mice. Reconstitution of RAG-2 KO mice with CD8+ T cells alone, or B cells alone, failed to restore FMN survival levels (65%+/-1.5 and 63%+/-1.0, respectively). It is concluded that, of the population of FMN that do not survive injury, CD4+ T lymphocytes, but not CD8+ T lymphocytes or B cells, mediate FMN survival after peripheral nerve injury.

Shank, J. R., S. J. Morgan, et al. (2003). "Bilateral peroneal nerve palsy following emergent stabilization of a pelvic ring injury." J Orthop Trauma 17(1): 67-70.
 External noninvasive compressive devices are becoming popular for emergent stabilization of pelvic ring disruptions. The ease of application utilizing available materials such as sheets, the noninvasive nature of such measures, and perceived absence of complications has made this a popular stabilization modality. The authors report a case of bilateral peroneal nerve palsy related to the use of external compressive wraps in a patient with pelvic ring injury.

Shaw, D. and M. S. Shoichet (2003). "Toward spinal cord injury repair strategies: peptide surface modification of expanded poly(tetrafluoroethylene) fibers for guided neurite outgrowth in vitro." J Craniofac Surg 14(3): 308-16.
 Expanded poly(tetrafluoroethylene) fibers were surface modified using an ultraviolet-activated mercury/ammonia reaction to yield amine-functional groups for the coupling of laminin-derived cell adhesive peptides CYIGSR, CDPGYIGSR, CIKVAV, and CQAASIKVAV. Surface elemental composition, determined by X-ray photoelectron spectroscopy, and radiolabeling data indicated that the amount of peptide introduced was approximately equivalent regardless of peptide type, yet mixed peptide surfaces had approximately 60% YIGSR and 40% IKVAV. The peptide-modified surfaces were compared in terms of the response of dorsal root ganglia with neurite length and number of cells attached to each fiber measured. All peptide-functionalized surfaces had a greater cellular response than the aminated ePTFE and ePTFE controls. Surfaces modified with extended peptide sequences CDPGYIGSR and CQAASIKVAV demonstrated a greater cellular response than those modified with the shorter peptide sequences CYIGSR and CIKVAV, respectively, likely because the extended peptides more closely mimic the three-dimensional conformation that the peptides maintain in laminin. Differences in neurite extension were evident among the peptide-functionalized surfaces, with the longest neurites observed on surfaces modified with both CQAASIKVAV and CDPGYIGSR. The "guidance capacity" of the fibers as a function of fiber diameter was investigated in terms of length and directionality of neurite outgrowth. As fiber diameter decreased (from 100+ to 10 microm), the neurites tended to grow to a greater degree down the length of the fiber. The thinnest fibers (with diameters <20 microm) extended shorter neurites than the fibers with a wider diameter. Combining neurite length with guidance indicated that of the fiber diameters investigated, the optimal fiber diameter for neurite guidance was between 30 and 50 microm.

Shen, A., D. Zhu, et al. (2003). "Increased gene expression of beta-1,4-galactosyltransferase I in rat injured sciatic nerve." J Mol Neurosci 21(2): 103-10.
 During neurite outgrowth on basal lamina, cell-surface beta-1,4-galactosyltransferase I (beta-1,4-GalT-I) functions as one of the receptors of laminin by binding to N-linked oligosaccharides on the laminin E8 domain. In the present study, it was revealed that in rat injured sciatic nerves, the expression of beta-1,4-GalT-I mRNA reached its peak 2-3 d after axotomy in both proximal and distal stumps, and decreased thereafter as demonstrated by Northern blot analysis. In situ hybridization revealed that beta-1,4-GalT-I mRNAmainly localized in Schwann cells of the injured nerves. Moreover the Galbeta1-4GlcNAc (N-acetylglucosamine) group mainly localized in Schwann cells of the injured nerves by Ricinus communis agglutinin-I (RCA-I) lectin histochemistry. However, the changes in abundance of the Galbeta1-4GlcNAc group in injured nerves were not consistent with the expression of beta-1, 4-GalT-I mRNA. These findings indicate that beta-1,4-GalT-I might be involved in the regeneration of injured peripheral nerves at the early injury stage.

Shen, A., J. Yan, et al. (2003). "Overexpression of beta-1,4-galactosyltransferase I in rat Schwann cells promotes the growth of co-cultured dorsal root ganglia." Neurosci Lett 342(3): 159-62.
 The cell surface beta-1,4-galactosyltransferase I (beta-1,4-GalT-I) functions as one of the receptors of laminin during the neurite outgrowth on basal lamina by binding to N-linked oligosaccharides in the laminin E8 domain. In this study, we demonstrated that the purified rat Schwann cells transfected with the expression plasmid of beta-1,4-GalT-I cDNA transiently promoted outgrowth and elongation of the neurites from co-cultured rat dorsal root ganglia, while those transfected with the antisense expression plasmid of beta-1,4-GalT-I had the opposite effects. These results suggested that the expression of beta-1,4-GalT-I in Schwann cells of peripheral nerve might promote both growth of developmental neuron and regeneration of injured nerve.

Shibuya, S., O. Miyamoto, et al. (2003). "Temporal progressive antigen expression in radial glia after contusive spinal cord injury in adult rats." Glia 42(2): 172-83.
 In the development of the CNS, radial glial cells are among the first cells derived from neuroepithelial cells. Recent studies have reported that radial glia possess properties of neural stem cells. We analyzed the antigen expression and distribution of radial glia after spinal cord injury (SCI). Sprague-Dawley rats had a laminectomy at Th11-12, and spinal cord contusion was created by compression with 30 g of force for 10 min. In the injury group, rats were examined at 24 h and 1, 4, and 12 weeks after injury. Frozen sections of 20-microm thickness were prepared from regions 5 and 10 mm rostral and caudal to the injury epicenter. Immunohistochemical staining was performed using antibodies to 3CB2 (a specific marker for radial glia), nestin, and glial fibrillary acidic protein (GFAP). At 1 week after injury, radial glia that bound anti-3CB2 MAb had spread throughout the white matter from below the pial surface. From 4 weeks after injury, 3CB2 expression was also observed in the gray matter around the central canal, and was especially strong around the ependymal cells and around blood vessels. In double-immunohistochemical assays for 3CB2 and GFAP or 3CB2 and nestin, coexpression was observed in subpial structures that extended into the white matter as arborizing processes and around blood vessels in the gray matter. The present study demonstrated the emergence of radial glia after SCI in adult mammals. Radial glia derived from subpial astrocytes most likely play an important role in neural repair and regeneration after SCI.

Shimazaki, K., K. Yoshida, et al. (2003). "Cytokines regulate c-Met expression in cultured astrocytes." Brain Res 962(1-2): 105-10.
 We investigated c-Met expression in cultured astrocytes and their regulation by cytokines. Immunocytochemistry revealed that c-Met was expressed in cultured astrocytes. Western blotting revealed that acidic and basic fibroblast growth factor (FGF) enhanced and hepatocyte growth factor (HGF) reduced c-Met expression. Reverse transcription-polymerase chain reaction revealed that FGFs and HGF enhanced c-met expression. These findings suggest that c-Met expressed in astrocytes may have important roles during the nervous system regeneration.

Shin, D. H., E. Lee, et al. (2003). "Growth-associated protein-43 is elevated in the injured rat sciatic nerve after low power laser irradiation." Neurosci Lett 344(2): 71-4.
 Low power laser irradiation (LPLI) has been used in the treatment of peripheral nerve injury. In this study, we verified its therapeutic effect on neuronal regeneration by finding elevated immunoreactivities (IRs) of growth-associated protein-43 (GAP-43), which is up-regulated during neuronal regeneration. Twenty Sprague-Dawley rats received a standardized crush injury of the sciatic nerve, mimicking the clinical situations accompanying partial axonotmesis. The injured nerve received calculated LPLI therapy immediately after injury and for 4 consecutive days thereafter. The walking movements of the animals were scored using the sciatic functional index (SFI). In the laser treated rats, the SFI level was higher in the laser treated animals at 3-4 weeks while the SFIs of the laser treated and untreated rats reached normal levels at 5 weeks after surgery. In immunocytochemical study, although GAP-43 IRs increased both in the untreated control and the LPLI treated groups after injury, the number of GAP-43 IR nerve fibers was much more increased in the LPLI group than those in the control group. The elevated numbers of GAP-43 IR nerve fibers reached a peak 3 weeks after injury, and then declined in both the untreated control and the LPLI groups at 5 weeks, with no differences in the numbers of GAP-43 IR nerve fibers of the two groups at this stage. This immunocytochemical study using GAP-43 antibody study shows for the first time that LPLI has an effect on the early stages of the nerve recovery process following sciatic nerve injury.

Siconolfi, L. B. and N. W. Seeds (2003). "Mice lacking tissue plasminogen activator and urokinase plasminogen activator genes show attenuated matrix metalloproteases activity after sciatic nerve crush." J Neurosci Res 74(3): 430-4.
 Plasminogen activators (PAs), tissue PA (tPA) and urokinase PA (uPA), have been shown to be induced in sensory neurons after sciatic nerve crush. These findings suggested that PAs facilitate peripheral nerve regeneration by digesting adhesive cell contacts and by activation of other proteases, thereby initiating a proteolytic cascade. Both tPA and uPA activate some matrix metalloproteases (MMPs), indirectly via plasminogen activation or directly, such as the uPA activation of MMP-2. In this study, we demonstrated, by using tPA and uPA knockout mice, that a lack of a plasminogen activator affected MMP-9 and MMP-2 activity after crushing of the sciatic nerve. These findings show that the PAs are important for MMP-9 and MMP-2 activity at the crush site.

Sicotte, M., O. Tsatas, et al. (2003). "Immunization with myelin or recombinant Nogo-66/MAG in alum promotes axon regeneration and sprouting after corticospinal tract lesions in the spinal cord." Mol Cell Neurosci 23(2): 251-63.
 We have shown previously that immunization with myelin in incomplete Freund's adjuvant (IFA) is able to promote robust regeneration of corticospinal tract fibers in adult mice. In the present study the effectiveness of such immunization with myelin was compared to that of a combination of two axon growth inhibitors in myelin, Nogo-66 (the 66-amino-acid inhibitory region of Nogo-A) and myelin-associated glycoprotein (MAG). The effectiveness of two adjuvants, IFA and aluminum hydroxide (Alum), was also compared, the latter being one that can be used in humans. In addition, larger dorsal overhemisections were made at the lower thoracic level, which resulted in a larger scar. These studies were carried out in SJL/J mice, a mouse strain that is susceptible to autoimmune experimental allergic encephalomyelitis (EAE). None of the immunized mice developed EAE. Long-distance axon regeneration and sprouting of the corticospinal tract was seen in myelin and Nogo-66/MAG immunized mice. Alum was as effective or better than IFA as the adjuvant. Overall, the robustness of axon growth and sprouting was greater in mice immunized with myelin. The abundance of this growth was less than in our earlier work in which smaller lesions were made, pointing to the possible influence of inhibitors in the scar. This work shows, however, that axon growth inhibitors in myelin can be selectively blocked using this immunization approach to promote long-distance axon regeneration in the spinal cord.

Silani, V. and N. Leigh (2003). "Stem therapy for ALS: hope and reality." Amyotroph Lateral Scler Other Motor Neuron Disord 4(1): 8-10.
 All are agreed that there is pressing need for an effective treatment for Amyotrophic Lateral Sclerosis (ALS; MND). Such treatment may derive from a combination of therapeutic strategies aimed at different aspects of the disorder, and might include drugs directed at the initial, intermediate or terminal cascade of events leading to cell death, as well as the use of stem cells to replace dead motor neurons, or to protect those that remain. The attraction of cell implantation or transplantation is that it might help to overcome the inability of the CNS to replace lost neurons. It is also clear that neural implantation will yield little benefit if the donor cells fail to integrate functionally into the recipient CNS circuitry. In this respect, ALS poses an especially difficult problem. The recent breakthroughs in stem cell research might nevertheless provide possibilities for neural implantation and cell replacement therapy for patients with ALS. The potential impact of these new approaches to neurodegenerative diseases has been emphasised by the many experiments using human foetal cell grafts in patients affected by Parkinson's and Huntington's disease. Clinical benefits in Parkinson's disease seem to be associated with integration of the donor cells into the recipient brain. Despite promising results, however, significant constraints have hampered the use of foetal cells for neural implantation and transplantation. Besides ethical concerns, the viability, purity, and final destiny of the foetal tissue have not been completely defined. Foetal cells are, in addition, post-mitotic and cannot be expanded or stored for long periods, necessitating close synchronisation of tissue donation and neurosurgery.

Sima, A. A. (2003). "C-peptide and diabetic neuropathy." Expert Opin Investig Drugs 12(9): 1471-88.
 Diabetic polyneuropathy (DPN) is the most common chronic complication of diabetes and affects Type 1 diabetic patients disproportionately. In the last two decades it has become increasingly evident that underlying metabolic, molecular and functional mechanisms and, ultimately, structural changes differ in DPN between the two major types of diabetes. In Type 1 diabetes, impaired insulin/C-peptide action has emerged as a prominent pathogenetic factor. C-peptide was long considered to be biologically inactive. During the last number of years it has been shown to have a number of insulin-like effects but without affecting blood glucose levels. Preclinical studies have demonstrated effects on Na(+)/K(+)-ATPase activity, endothelial nitric oxide synthase, expression of neurotrophic factors and regulation of molecular species underlying the degeneration of the nodal apparatus in Type 1 diabetic nerves, as well as DNA binding of transcription factors and modulation of apoptotic phenomena. In animal studies, these effects have translated into protection and improvement of functional abnormalities, promotion of nerve fibre regeneration, protection of structural changes and amelioration of apoptotic phenomena targeting central and peripheral nerve cell constituents. Several small-scale clinical trials confirm these beneficial effects on autonomic and somatic nerve function and blood flow in a variety of tissues. Therefore, evidence to date indicating that replacement of C-peptide in patients with Type 1 diabetes will retard and prevent chronic complication is real and encouraging. Large-scale clinical trials necessary to bring this natural substance into the clinical arena should, therefore, be encouraged and accelerated.

Simonen, M., V. Pedersen, et al. (2003). "Systemic deletion of the myelin-associated outgrowth inhibitor Nogo-A improves regenerative and plastic responses after spinal cord injury." Neuron 38(2): 201-11.
 To investigate the role of the myelin-associated protein Nogo-A on axon sprouting and regeneration in the adult central nervous system (CNS), we generated Nogo-A-deficient mice. Nogo-A knockout (KO) mice were viable, fertile, and not obviously afflicted by major developmental or neurological disturbances. The shorter splice form Nogo-B was strongly upregulated in the CNS. The inhibitory effect of spinal cord extract for growing neurites was decreased in the KO mice. Two weeks following adult dorsal hemisection of the thoracic spinal cord, Nogo-A KO mice displayed more corticospinal tract (CST) fibers growing toward and into the lesion compared to their wild-type littermates. CST fibers caudal to the lesion-regenerating and/or sprouting from spared intact fibers-were also found to be more frequent in Nogo-A-deficient animals.

Singru, P. S., A. J. Sakharkar, et al. (2003). "Neuronal nitric oxide synthase in the olfactory system of an adult teleost fish Oreochromis mossambicus." Brain Res 977(2): 157-68.
 The aim of the present study is to explore the distribution of nitric oxide synthase in the olfactory system of an adult teleost, Oreochromis mossambicus using neuronal nitric oxide synthase (nNOS) immunocytochemistry and nicotinamide adenine dinucleotide phosphate diaphorase (NADPHd) histochemistry methods. Intense nNOS immunoreactivity was noticed in several olfactory receptor neurons (ORNs), in their axonal extensions over the olfactory nerve and in some basal cells of the olfactory epithelium. nNOS containing fascicles of the ORNs enter the bulb from its rostral pole, spread in the olfactory nerve layer in the periphery of the bulb and display massive innervation of the olfactory glomeruli. Unilateral ablation of the olfactory organ resulted in dramatic loss of nNOS immunoreactivity in the olfactory nerve layer of the ipsilateral bulb. In the olfactory bulb of intact fish, some granule cells showed intense immunoreactivity; dendrites arising from the granule cells could be traced to the glomerular layer. Of particular interest is the occurrence of nNOS immunoreactivity in the ganglion cells of the nervus terminalis. nNOS containing fibers were also encountered in the medial olfactory tracts as they extend to the telencephalon. The NADPHd staining generally coincides with that of nNOS suggesting that it may serve as a marker for nNOS in the olfactory system of this fish. However, mismatch was encountered in the case of mitral cells, while all are nNOS-negative, few were NADPHd positive. The present study for the first time revealed the occurrence of nNOS immunoreactivity in the ORNs of an adult vertebrate and suggests a role for nitric oxide in the transduction of odor stimuli, regeneration of olfactory epithelium and processing of olfactory signals.

Skundric, D. S. and R. P. Lisak (2003). "Role of neuropoietic cytokines in development and progression of diabetic polyneuropathy: from glucose metabolism to neurodegeneration." Int J Exp Diabesity Res 4(4): 303-12.
 Diabetic neuropathy develops as a result of hyperglycemia-induced local metabolic and microvascular changes in both type I and type II diabetes mellitus. Diabetic neuropathy shows slower impulse conduction, axonal degeneration, and impaired regeneration. Diabetic neuropathy affects peripheral, central, and visceral sensorimotor and motor nerves, causing improper locomotor and visceral organ dysfunctions. The pathogenesis of diabetic neuropathy is complex and involves multiple pathways. Lack of success in preventing neuropathy, even with successful treatment of hyperglycemia, suggests the presence of early mediators between hyperglycemia-induced metabolic and enzymatic changes and functional and structural properties of Schwann cells (SCs) and axons. It is feasible that once activated, such mediators can act independently of the initial metabolic stimulus to modulate SC-axonal communication. Neuropoietic cytokines, including interleukin-1 (IL-1), interleukin-6 (IL-6), leukemia inhibitory factor (LIF), ciliary neurotrophic factor (CNTF), tumor necrosis factor alpha (TNF-alpha), and transforming growth factor beta (TGF-beta), exhibit pleiotrophic effects on homeostasis of glia and neurons in central, peripheral, and autonomic nervous system. These cytokines are produced locally by resident and infiltrating macrophages, lymphocytes, mast cells, SCs, fibroblasts, and sensory neurons. Metabolic changes induced by hyperglycemia lead to dysregulation of cytokine control. Moreover, their regulatory roles in nerve degeneration and regeneration may potentially be utilized for the prevention and/or therapy of diabetic neuropathy.

Smit, X., B. Stefan de Kool, et al. (2003). "Magnetoneurography: recording biomagnetic fields for quantitative evaluation of isolated rat sciatic nerves." J Neurosci Methods 125(1-2): 59-63.
 Magnetoneurography (MNG) is a technique to record the biomagnetic action fields of peripheral nerves. The benefits of MNG in contrast to electroneurography include the decreased signal disturbance caused by surrounding biological tissues and the use of a calibration pulse, both of which contribute to high reproducibility. MNG has proven to be a valuable tool to quantitate peripheral nerve regeneration in rabbits. However, the most commonly used model to study the peripheral nervous system is the rat sciatic nerve. Until now, the small size of the nerve impeded accurate MNG measurements in rat. This report describes a custom made recording chamber that allows accurate control of conduction distances and temperature and enables adequate MNG measurements of isolated sciatic nerves of Wistar rats. We applied biphasic stimulation with optimized grounding to reduce the stimulus artefact. A high reproducibility of signals was demonstrated. 'Ex vivo' nerve viability was assured for at least 2 h after dissection. In conclusion, MNG is a powerful tool to quantitatively evaluate the function of rat sciatic nerves and will be used for the early assessment of nerve regeneration.

Smittkamp, S. E., D. L. Park, et al. (2003). "Effects of age and cochlear damage on the metabolic activity of the avian cochlear nucleus." Hear Res 175(1-2): 101-11.
 Most aging commercially raised broiler chickens display a progressive loss of cochlear hair cells in a pattern similar to the cochlear degeneration found in aging humans: basal (high frequency) hair cells are affected first, followed by apical (low frequency) hair cells [Durham et al., Hear. Res. 166 (2002) 82-95]. Here, cochlear anatomy was assessed from scanning electron micrographs. Then, the metabolic activity of cochlear nucleus (nucleus magnocellularis, NM) neurons in 15-19, 30, 39, 40, and 65-66 week old broiler chickens was examined using cytochrome oxidase histochemistry and compared to the degree of cochlear abnormality. Cochleae of 15-19 week old birds are largely normal; therefore the level of NM metabolic activity is considered the baseline. Cochleae of the 30 week old group display mild damage and hair cell regeneration in the base. Metabolic activity in rostral (high frequency) NM is increased relative to the baseline, while activity remains unchanged in caudal (low frequency) NM. The 39 and 65-66 week old groups display severe and total damage extending into the apex of the cochlea. Metabolic activity is decreased in rostral and caudal NM at these ages. These results suggest that auditory central nervous system metabolism (cytochrome oxidase activity) is affected by changes in the aging chicken cochlea.

Sobol, J. B., I. J. Lowe, et al. (2003). "Effects of delaying FK506 administration on neuroregeneration in a rodent model." J Reconstr Microsurg 19(2): 113-8.
 FK506 is an immunosuppressant drug that has been shown experimentally to stimulate nerve growth and speed functional recovery, when administered immediately after peripheral nerve injury. However, the clinical scenario of a peripheral nerve injury is often associated with either a delayed diagnosis or reconstruction. The purpose of this study was to determine the efficacy of FK506 on neuroregeneration with delayed administration. Thirty-two Lewis rats underwent tibial nerve transection with immediate repair. Animals were left untreated, or were treated with daily injections of FK506 (2 mg/kg), started on the day of surgery, postoperative day 3, or postoperative day 5. Animals underwent walking track analysis to assess functional nerve recovery. Nerves were harvested for histomorphometric analysis on postoperative days 21, 28, and 42. Histomorphometry demonstrated that all treatment groups, regardless of the time of drug initiation, demonstrated evidence of enhanced neuroregeneration, compared to the untreated group. Histomorphometric data from groups harvested on day 21 demonstrated a statistically significant improvement in neuroregeneration in the immediate and 3-day delay groups. Therefore, the beneficial effects of FK506 on neuroregeneration are not restricted to immediate administration, but these effects significantly diminish when FK506 is administered 3 days after nerve injury.

Sonmez, A., M. Bayramicli, et al. (2003). "Reconstruction of the weight-bearing surface of the foot with nonneurosensory free flaps." Plast Reconstr Surg 111(7): 2230-6.
 Neurotized fasciocutaneous flaps and split-skin grafted muscle flaps are the most frequently used free flap alternatives for the reconstruction of weight-bearing surfaces of the foot. An objective comparison of the innate characteristics of these two flap types, with respect to long-term stability, has not been possible because sensory reinnervation in the fasciocutaneous flaps has been a confounding factor. This study compares nonsensate fasciocutaneous flaps (n = 9) with nonsensate split-skin grafted muscle flaps (n = 11), with mean follow-up periods of 34.3 and 31.3 months, respectively. Patients completed a form that included questions regarding degree of pain at the operative site, presence of ulcers, ability to wear normal shoes, employment status, and time spent standing on foot. Touch and deep sensation were evaluated with Semmes-Weinstein and vibration tests, respectively. Significantly less pain and less ulceration (p < 0.05) were observed in the fasciocutaneous group. Semmes-Weinstein monofilament tests revealed poorer results with split-skin grafted muscle flaps, compared with fasciocutaneous flaps. These results indicate that even if the sensory protection of fasciocutaneous flaps is not considered, these flaps have superior properties, compared with split-skin grafted muscle flaps.

Sorci, G., F. Riuzzi, et al. (2003). "S100B inhibits myogenic differentiation and myotube formation in a RAGE-independent manner." Mol Cell Biol 23(14): 4870-81.
 S100B is a Ca(2+)-modulated protein of the EF-hand type with both intracellular and extracellular roles. S100B, which is most abundant in the brain, has been shown to exert trophic and toxic effects on neurons depending on the concentration attained in the extracellular space. S100B is also found in normal serum, and its serum concentration increases in several nervous and nonnervous pathological conditions, suggesting that S100B-expressing cells outside the brain might release the protein and S100B might exert effects on nonnervous cells. We show here that at picomolar to nanomolar levels, S100B inhibits myogenic differentiation of rat L6 myoblasts via inactivation of p38 kinase with resulting decrease in the expression of the myogenic differentiation markers, myogenin, muscle creatine kinase, and myosin heavy chain, and reduction of myotube formation. Although myoblasts express the multiligand receptor RAGE, which has been shown to transduce S100B effects on neurons, S100B produces identical effects on myoblasts overexpressing either full-length RAGE or RAGE lacking the transducing domain. This suggests that S100B affects myoblasts by interacting with another receptor and that RAGE is not the only receptor for S100B. Our data suggest that S100B might participate in the regulation of muscle development and regeneration by inhibiting crucial steps of the myogenic program in a RAGE-independent manner.

Soto, I., B. Marie, et al. (2003). "FGF-2 modulates expression and distribution of GAP-43 in frog retinal ganglion cells after optic nerve injury." J Neurosci Res 73(4): 507-17.
 Basic fibroblast growth factor (bFGF or FGF-2) has been implicated as a trophic factor that promotes survival and neurite outgrowth of neurons. We found previously that application of FGF-2 to the proximal stump of the injured axon increases retinal ganglion cell (RGC) survival. We determine here the effect of FGF-2 on expression of the axonal growth-associated phosphoprotein (GAP)-43 in retinal ganglion cells and tectum of Rana pipiens during regeneration of the optic nerve. In control retinas, GAP-43 protein was found in the optic fiber layer and in optic nerve; mRNA levels were low. After axotomy, mRNA levels increased sevenfold and GAP-43 protein was significantly increased. GAP-43 was localized in retinal axons and in a subset of RGC cell bodies and dendrites. This upregulation of GAP-43 was sustained through the period in which retinal axons reconnect with their target in the tectum. FGF-2 application to the injured nerve, but not to the eyeball, increased GAP-43 mRNA in the retina but decreased GAP-43 protein levels and decreased the number of immunopositive cell bodies. In the tectum, no treatment affected GAP-43 mRNA but FGF-2 application to the axotomized optic nerve increased GAP-43 protein in regenerating retinal projections. We conclude that FGF-2 upregulates the synthesis and alters the distribution of the axonal growth-promoting protein GAP-43, suggesting that it may enhance axonal regrowth.

Spencer, T., M. Domeniconi, et al. (2003). "New roles for old proteins in adult CNS axonal regeneration." Curr Opin Neurobiol 13(1): 133-9.
 The past year has yielded many insights and a few surprises in the field of axonal regeneration. The identification of oligodendrocyte-myelin glycoprotein as an inhibitor of axonal growth, and the discovery that the three major myelin-associated inhibitors of CNS regeneration share the same functional receptor, has launched a new wave of studies that aim to identify the signaling components of these inhibitory pathways. These findings also offer new avenues of research directed toward blocking possible therapeutic targets that inhibit regeneration and toward encouraging axonal regeneration in the CNS after injury.

Spira, M. E., R. Oren, et al. (2003). "Critical calpain-dependent ultrastructural alterations underlie the transformation of an axonal segment into a growth cone after axotomy of cultured Aplysia neurons." J Comp Neurol 457(3): 293-312.
 The transformation of a stable axonal segment into a motile growth cone is a critical step in the regeneration of amputated axons. In earlier studies we found that axotomy of cultured Aplysia neurons leads to a transient and local elevation of the free intracellular Ca2+ concentration, resulting in calpain activation, localized proteolysis of submembranal spectrin, and, eventually, growth cone formation. Moreover, inhibition of calpain by calpeptin prior to axotomy inhibits growth cone formation. Here we investigated the mechanisms by which calpain activation participates in the transformation of an axonal segment into a growth cone. To that end we compared the ultrastructural alterations induced by axotomy performed under control conditions with those caused by axotomy performed in the presence of calpeptin, using cultured Aplysia neurons as a model. We identified the critical calpain-dependent cytoarchitectural alterations that underlie the formation of a growth cone after axotomy. Calpain-dependent processes lead to restructuring of the neurofilaments and microtubules to form an altered cytoskeletal region 50-150 microm proximal to the tip of the transected axon in which vesicles accumulate. The dense pool of vesicles forms in close proximity to a segment of the plasma membrane along which the spectrin membrane skeleton has been proteolyzed by calpain. We suggest that the rearrangement of the cytoskeleton forms a transient cellular compartment that traps transported vesicles and serves as a locus for microtubule polymerization. We propose that this cytoskeletal configuration facilitates the fusion of vesicles with the plasma membrane, promoting the extension of the growth cone's lamellipodium. The growth process is further supported by the radial polymerization of microtubules from the growth cone's center.

St John, J. A., H. J. Clarris, et al. (2003). "Sorting and convergence of primary olfactory axons are independent of the olfactory bulb." J Comp Neurol 464(2): 131-40.
 Primary olfactory axons expressing the same odorant receptor gene sort out and converge to fixed sites in the olfactory bulb. We examined the guidance of axons expressing the P2 odorant receptor when they were challenged with different cellular environments in vivo. In the mutant extratoes mouse, the olfactory bulb is lacking and is replaced by a fibrocellular mass. In these animals, primary olfactory axons form glomerular-like loci despite the absence of normal postsynaptic targets. P2 axons are able to sort out from other axons in this fibrocellular mass and converge to form loci of like axons. The sites of these loci along mediolateral and ventrodorsal axes were highly variable. Similar convergence was observed for larger subpopulations of axons expressing the same cell surface carbohydrates. The sorting out and convergence of like axons also occurred during regeneration following bulbectomy. Olfactory axon behaviour in these models demonstrates that sorting and convergence of axons are independent of the target, which instead provides distinct topographic cues for guidance.

St John, S. J., M. Garcea, et al. (2003). "The time course of taste bud regeneration after glossopharyngeal or greater superficial petrosal nerve transection in rats." Chem Senses 28(1): 33-43.
 We previously have published data detailing the time course of taste bud regeneration in the anterior tongue following transection of the chorda tympani (CT) nerve in the rat. This study extends the prior work by determining the time course of taste bud regeneration in the vallate papilla, soft palate and nasoincisor ducts (NID) following transection of either the glossopharyngeal (GL) or greater superficial petrosal (GSP) nerve. Following GL transection in rats (n = 6 per time point), taste buds reappeared in the vallate papilla between 15 and 28 days after surgery, and returned to 80.3% of control levels (n = 12) of taste buds by 70 days postsurgery. The first appearance and the final percentage of the normal complement of regenerated vallate taste buds after GL transection resembled that seen previously in the anterior tongue after CT transection. However, in the latter case, regenerated taste buds reached asymptotic levels by 42 days after surgery, whereas within the time frame of the present study, a clear asymptotic return of vallate taste buds was not observed. In contrast to the posterior (and anterior) tongue, only 25% of the normal complement of palatal taste buds regenerated by 112 days and 224 days after GSP transection (n = 9). The difference in regenerative capacity might relate to the surgical approach used to transect the GSP. These experiments provide useful parametric data for investigators studying the functional consequences of gustatory nerve transection and regeneration.

Stanic, D., D. I. Finkelstein, et al. (2003). "Timecourse of striatal re-innervation following lesions of dopaminergic SNpc neurons of the rat." Eur J Neurosci 18(5): 1175-88.
 Previously we described the extent of sprouting that axons of the rat substantia nigra pars compacta (SNpc) undergo to grow new synapses and re-innervate the dorsal striatum 16 weeks after partial lesions. Here we provide insights into the timing of events related to the re-innervation of the dorsal striatum by regenerating dopaminergic nigrostriatal axons over a 104-week period after partial SNpc lesioning. Density of dopamine transporter and tyrosine hydroxylase immunoreactive axonal varicosities (terminals) decreased up to 80% 4 weeks after lesioning but returned to normal by 16 weeks, unless SNpc lesions were greater than 75%. Neuronal tracer injections into the SNpc revealed a 119% increase in axon fibres (4 mm rostral to the SNpc) along the medial forebrain bundle 4 weeks after lesioning. SNpc cells underwent phenotypic changes. Four weeks after lesioning the proportion of SNpc neurons that expressed tyrosine hydroxylase fell from 90% to 38% but returned to 78% by 32 weeks. We discuss these phenotype changes in the context of neurogenesis. Significant reductions in dopamine levels in rats with medium (30-75%) lesions returned to normal by 16 weeks whereas recovery was not observed if lesions were larger than 75%. Finally, rotational behaviour of animals in response to amphetamine was examined. The clear rightward turning bias observed after 2 weeks recovered by 16 weeks in animals with medium (30-75%) lesions but was still present when lesions were larger. These studies provide insights into the processes that regulate sprouting responses in the central nervous system following injury.

Stanic, D., C. L. Parish, et al. (2003). "Changes in function and ultrastructure of striatal dopaminergic terminals that regenerate following partial lesions of the SNpc." J Neurochem 86(2): 329-43.
 Following partial substantia nigra lesions, remaining dopaminergic neurones sprout, returning terminal density in the dorsal striatum to normal by 16 weeks. This suggests regeneration and maintenance of terminal density is regulated to release appropriate levels of dopamine. This study examined the structure and function of these reinnervated terminals, defining characteristics of dopamine uptake and release, density and affinity of the dopamine transporter (DAT) and ultrastructural morphology of dopamine terminals in the reinnervated dorsal striatum. Finally, rotational behaviour of animals in response to amphetamine was examined 4 and 16 weeks after substantia nigra pars compacta (SNpc) lesions. Dopamine transport was markedly reduced 16 weeks after lesioning along with reduced density and affinity of DAT. Rate of dopamine release and peak concentration, measured electrochemically, was similar in lesioned and control animals, while clearance was prolonged after lesioning. Ultrastructurally, terminals after lesioning were morphologically distinct, having increased bouton size, vesicle number and mitochondria, and more proximal contacts on post-synaptic cells. After 4 weeks, tendency to rotate in response to amphetamine was proportional to lesion size. By 16 weeks, rotational behaviour returned to near normal in animals where lesions were less than 70%, although some animals demonstrated unusual rotational patterns at the beginning and end of the amphetamine effect. Together, these changes indicate that sprouted terminals are well compensated for dopamine release but that transport mechanisms are functionally impaired. We discuss these results in terms of implications for dyskinesia and other behavioural states.

Starowicz, K. and B. Przewlocka (2003). "The role of melanocortins and their receptors in inflammatory processes, nerve regeneration and nociception." Life Sci 73(7): 823-47.
 The melanocortins are a family of bioactive peptides derived from proopiomelanocortin. Those peptides, included among hormones and comprising ACTH, alpha-MSH, beta-MSH and gamma-MSH, are best known mainly for their physiological effects, such as the control of skin pigmentation by alpha-MSH, and ACTH effects on pigmentation and steroidogenesis. Melanocortins are released in various sites in the central nervous system and in peripheral tissues, and participate in the regulation of multiple physiological functions. They are involved in grooming behavior, food intake and thermoregulation processes, and can also modulate the response of the immune system in inflammatory states. Research of the past decade provided evidence that melanocortins could elicit their diverse biological effects by binding to a distinct family of G protein-coupled receptors with seven transmembrane domains. To date, five melanocortin receptor genes have been cloned and characterized. Those receptors differ in their tissue distribution and in their ability to recognize various melanocortins. These advances have opened up new horizons for exploring the significance of melanocortins, their ligands and their receptors for a variety of important physiological functions. We reviewed the origin of MSH peptides, the function and distribution of melanocortin receptors and their endogenous and exogenous ligands and the role of melanocortins and their receptors in inflammatory processes, nerve regeneration and nociception. Moreover, we analyzed their interaction with opioid peptides and finally, we discussed the postulated role of the melanocortin system in pain transmission at the spinal cord level.

Stedtfeld, H. W., W. Attmanspacher, et al. (2003). "[Fixation of humeral head fractures with antegrade intramedullary nailing]." Zentralbl Chir 128(1): 6-11.
 The new method of antegrade intramedullary fixation of humeral head fractures is based on a straight proximal humeral nail with special head fixation screws and conventional interlocking screws at the proximal end of the shaft fragment leaving an axillary nerve shelter space in between. The nail acts as a central load carrier. The head fixation screws run through threaded holes in the proximal end of the nail thus being held in a stiff angle and without gliding. The entry points of these screws correspond to the anatomical main portions of the lesser and greater tubercle. They allow a three-dimensional screw grip to the subchondral bony layer of the head fragment. The purpose of this intramedullary construct is to keep the fracture stable at a grade which allows instant postoperative active exercise and which corresponds to the needs of mechanical tranquility in a predominantly endosteal healing area. In a prospective clinical study 45 patients could be followed up after 3, 6 and 12 months. We found an ongoing improvement of the postoperative results up to an average Constant Score of 85.7 pts after one year. The complication rate was 16 %. The main complication was the screw protrusion into the joint.

Steinkamp, M., I. Geerling, et al. (2003). "Glial-derived neurotrophic factor regulates apoptosis in colonic epithelial cells." Gastroenterology 124(7): 1748-57.
 BACKGROUND & AIMS: Ablation of the enteric glia leads to a fulminant hemorrhagic jejunoileitis. We hypothesized that glial-derived neurotrophic factor (GDNF) may be involved in mucosal protection of the gut. Therefore, we examined the regulation of GDNF and its receptor (GFR-alpha1) in colonic inflammation and its effects on colonic epithelial cell apoptosis. METHODS: The expression of GDNF and GFR-alpha1 was investigated in experimental colitis of rats and in human inflammatory bowel disease (IBD). GDNF-induced activation of Akt (protein kinase B [PKB]) and mitogen-activated protein kinase (MAPK) in the colonic epithelial cell lines HT-29 and SW480 was studied. Furthermore, the antiapoptotic potency of GDNF in SW480 cells was evaluated. RESULTS: GDNF was specifically up-regulated in experimental rat colitis and in IBD. In contrast, GFR-alpha1 was constitutively expressed in rat and human colonic epithelium. GDNF potently activated MAPK and Akt (PKB) in colonic epithelial cells. Moreover, GDNF strongly prevented apoptosis in SW480 cells. Our data show that GDNF-mediated protection against apoptosis depends on activation of the MAPK and phosphatidylinositol 3-kinase/Akt (PKB) pathways. CONCLUSIONS: GDNF is up-regulated in IBD and has strong antiapoptotic properties in colonic epithelial cells. This points to a novel role of the neurotrophic factor GDNF for mucosal protection and regeneration in IBD.

Stenzl, A. (2003). "[New surgical method in bladder paralysis. Contracting the latissimus muscle to urinate (interview by Dr. Judith Neumaier)]." MMW Fortschr Med 145(6): 12.
 
Steward, O., B. Zheng, et al. (2003). "False resurrections: distinguishing regenerated from spared axons in the injured central nervous system." J Comp Neurol 459(1): 1-8.
 Several recent studies report that axon regeneration can be induced in the mature mammalian nervous system by novel treatments or genetic manipulations. In assessing these reports, it is important to be mindful of the history of regeneration research, which is littered with the corpses of studies that reported regeneration that later proved incorrect. One important reason is the "spared axon conundrum," in which axons that survive a lesion are mistakenly identified as having regenerated. Here, we illustrate the problem and propose criteria that may be used to identify regenerated vs. spared axons, focusing on the injured spinal cord.

Stix, G. (2003). "Ultimate self-improvement." Sci Am 289(3): 44-5.
 
Storer, P. D., D. Dolbeare, et al. (2003). "Treatment of chronically injured spinal cord with neurotrophic factors stimulates betaII-tubulin and GAP-43 expression in rubrospinal tract neurons." J Neurosci Res 74(4): 502-11.
 Exogenous neurotrophic factors provided at a spinal cord injury site promote regeneration of chronically injured rubrospinal tract (RST) neurons into a peripheral nerve graft. The present study tested whether the response to neurotrophins is associated with changes in the expression of two regeneration-associated genes, betaII-tubulin and growth-associated protein (GAP)-43. Adult female rats were subjected to a right full hemisection lesion via aspiration of the C3 spinal cord. A second aspiration lesion was made 4 weeks later and gel foam saturated in brain-derived neurotrophic factor (BDNF), glial cell-line derived neurotrophic factor (GDNF), or phosphate-buffered saline (PBS) was applied to the lesion site for 60 min. Using in situ hybridization, RST neurons were examined for changes in mRNA levels of betaII-tubulin and GAP-43 at 1, 3, and 7 days after treatment. Based on analysis of gene expression in single cells, there was no effect of BDNF treatment on either betaII-tubulin or GAP-43 mRNA expression at any time point. betaII-Tubulin mRNA levels were enhanced significantly at 1 and 3 days in animals treated with GDNF relative to levels in animals treated with PBS. Treatment with GDNF did not affect GAP-43 mRNA levels at 1 and 3 days, but at 7 days there was a significant increase in mRNA expression. Interestingly, 7 days after GDNF treatment, the mean cell size of chronically injured RST neurons was increased significantly. Although GDNF and BDNF both promote axonal regeneration by chronically injured neurons, only GDNF treatment is associated with upregulation of betaII-tubulin or GAP-43 mRNA. It is not clear from the present study how exogenous BDNF stimulates regrowth of injured axons.

Street, V. A., C. L. Bennett, et al. (2003). "Mutation of a putative protein degradation gene LITAF/SIMPLE in Charcot-Marie-Tooth disease 1C." Neurology 60(1): 22-6.
 BACKGROUND: Charcot-Marie-Tooth (CMT) neuropathy is a heterogeneous group of inherited disorders of the peripheral nervous system. The authors recently mapped an autosomal dominant demyelinating form of CMT type 1 (CMT1C) to chromosome 16p13.1-p12.3. OBJECTIVE: To find the gene mutations underlying CMT1C. METHODS: The authors used a combination of standard positional cloning and candidate gene approaches to identify the causal gene for CMT1C. Western blot analysis was used to determine relative protein levels in patient and control lymphocyte extracts. Northern blotting was used to characterize gene expression in 1) multiple tissues; 2) developing sciatic nerve; and 3) nerve-crush and nerve-transection experiments. RESULTS: The authors identified missense mutations (G112S, T115N, W116G) in the LITAFgene (lipopolysaccharide-induced tumor necrosis factor-alpha factor) in three CMT1C pedigrees. LITAF, which is also referred to as SIMPLE, is a widely expressed gene encoding a 161-amino acid protein that may play a role in protein degradation pathways. The mutations associated with CMT1C were found to cluster, defining a domain of the LITAF protein having a critical role in peripheral nerve function. Western blot analysis suggested that the T115N and W116G mutations do not alter the level of LITAF protein in peripheral blood lymphocytes. The LITAF transcript is expressed in sciatic nerve, but its level of expression is not altered during development or in response to nerve injury. This finding is in stark contrast to that seen for other known genes that cause CMT1. CONCLUSIONS: Mutations in LITAF may account for a significant proportion of CMT1 patients with previously unknown molecular diagnosis and may define a new mechanism of peripheral nerve perturbation leading to demyelinating neuropathy.

Strelau, J. and K. Unsicker (2003). "Neuroregeneration." Adv Neurol 91: 95-100.
 
Su, H. X. and E. Y. Cho (2003). "Sprouting of axon-like processes from axotomized retinal ganglion cells induced by normal and preinjured intravitreal optic nerve grafts." Brain Res 991(1-2): 150-62.
 The failure of axonal regeneration in the mammalian central nervous system (CNS) is currently attributed to the glial environment of the lesion site which elaborates a multitude of inhibitory factors. Less attention has been paid to the potential of trophic support associated with the CNS, especially in relation to the status of the damaged CNS after an injury has been evoked. Using a grafting paradigm to implant an optic nerve (ON) segment into the vitreous, we have addressed how a prior damage of the ON before grafting influences its ability to stimulate retinal ganglion cells (RGCs) to sprout axon-like processes. Our results showed that a normal noninjured ON implanted intravitreally stimulated sprouting of RGCs, as revealed by sliver staining of the sprouting cells, as well as increasing the number of RGCs which express GAP-43. A prior crush injury of the ON 7 days before its implantation into the vitreous resulted in a significant decrease in its ability to stimulate RGC sprouting when the crush lesion segment was used as the graft, whereas grafts taken from segments proximal and distal to the lesion segment had potencies similar to that of the noninjured graft. Both astrocytes and oligodendrocytes were drastically reduced in number in the lesion segment graft, suggesting their involvement in the secretion of soluble trophic factors that may play a role in the sprouting and regeneration of damaged neurons.

Su, Y. K. and J. S. Elam (2003). "Proteoglycan regulation of goldfish retinal explant growth on optic tectal membranes." Brain Res Dev Brain Res 142(2): 169-75.
 Regenerating goldfish retinal explants cultured on poly-L-lysine overlaid with membranes isolated from 21-day regenerating 1/3 anterior optic tectum (Ant. OTec) exhibited extensive defasciculated neurite outgrowth. Heparatinase treatment of membranes caused the complete inhibition of neurite outgrowth on that substrate. Western blot analysis showed that the OTec membranes contain a 300 kDa heparan sulfate proteoglycan. Explants cultured on 21-day regenerating 1/3 Ant. OTec membranes in the presence of 1 mM beta-xyloside, an axonal proteoglycan synthesis inhibitor, showed a significant reduction in the number of neurites per explant and in the average neurite length. Taken all together, the present results provide evidence that a 300-kDa membrane HSPG present in the Ant. OTec is necessary for axonal outgrowth and that axonal PGs are involved in modulating outgrowth on 21-day regenerating 1/3 Ant. OTec membranes.

Sugaya, K. (2003). "Neuroreplacement therapy and stem cell biology under disease conditions." Cell Mol Life Sci 60(9): 1891-902.
 Recent advances in stem cell technology are expanding our ability to replace a variety of cells throughout the body. In the past, neurological diseases caused by the degeneration of neuronal cells were considered incurable because of a long-held 'truism'; neurons do not regenerate during adulthood. However, this statement has been challenged, and we have now found much evidence that the brain is indeed capable of regenerating neurons after maturing. Based on this new concept, researchers have shown neural differentiation of stem cells and recovery of function following transplantation of these cells into the brain. These results may promise a bright future for clinical applications of stem cell strategies in neurological diseases; however, we must consider the pathophysiological environments of individual diseases that may affect stem cell biology. Before we begin to develop clinical applications, we must consider environmental factors that have not been discussed in the current preclinical studies. Here, we study cases of Alzheimer's disease and schizophrenia and discuss the effects of environmental factors under disease conditions.

Sullivan, K. A., M. S. Brown, et al. (2003). "Digital electron microscopic examination of human sural nerve biopsies." J Peripher Nerv Syst 8(4): 260-70.
 Diabetic peripheral polyneuropathy is characterized by axonal degeneration and regeneration as well as by Schwann cell and microvascular changes. These changes have been described at both the light (LM) and the electron microscopic (EM) levels; however, EM has not been applied to large clinical trials. Our goal was to adapt the rigorous techniques used for quantifying human biopsies with LM image analysis to accommodate ultrastructural analyses. We applied digital image capture and analysis to the ultrastructural examination of axons in sural nerve biopsies from diabetic patients enrolled in a multicenter clinical trial. The selection of sural nerve biopsies was based on the quality of specimen fixation, absence of physical distortion, and nerve fascicle size (>/=100 000; </=425 000 micro m2). Thin sections were collected on formvar-coated slot grids, stabilized with carbon and scanned on a Phillips CM100 transmission electron microscope. Digital images were captured with a Kodak Megaplus 1.6 camera. A montage was constructed using software derived from aerial mapping applications, and this virtual image was viewed by EM readers. Computer-assisted analyses included identification and labeling of individual axons and axons within regenerating clusters. The average density of regenerating myelinated axon clusters per mm2 was 65.8 +/- 5.1, range of 0-412 (n = 193). These techniques increase the number of samples that may be analyzed by EM and extend the use of this technique to clinical trials using tissue biopsies as a primary endpoint.

Sundar, T. (2003). "[On the track of healing stem cells]." Tidsskr Nor Laegeforen 123(10): 1378-81.
 
Sun, Y., K. Jin, et al. (2003). "VEGF-induced neuroprotection, neurogenesis, and angiogenesis after focal cerebral ischemia." J Clin Invest 111(12): 1843-51.
 Vascular endothelial growth factor (VEGF) is an angiogenic protein with therapeutic potential in ischemic disorders, including stroke. VEGF confers neuroprotection and promotes neurogenesis and cerebral angiogenesis, but the manner in which these effects may interact in the ischemic brain is poorly understood. We produced focal cerebral ischemia by middle cerebral artery occlusion for 90 minutes in the adult rat brain and measured infarct size, neurological function, BrdU labeling of neuroproliferative zones, and vWF-immunoreactive vascular profiles, without and with intracerebroventricular administration of VEGF on days 1-3 of reperfusion. VEGF reduced infarct size, improved neurological performance, enhanced the delayed survival of newborn neurons in the dentate gyrus and subventricular zone, and stimulated angiogenesis in the striatal ischemic penumbra, but not the dentate gyrus. We conclude that in the ischemic brain VEGF exerts an acute neuroprotective effect, as well as longer latency effects on survival of new neurons and on angiogenesis, and that these effects appear to operate independently. VEGF may, therefore, improve histological and functional outcome from stroke through multiple mechanisms.

Sundine, M. J., E. E. Quan, et al. (2003). "The use of end-to-side nerve grafts to reinnervate the paralyzed orbicularis oculi muscle." Plast Reconstr Surg 111(7): 2255-64.
 Facial paralysis is a serious neurologic disorder, particularly when it affects the eye. Loss of the protective blink reflex may lead to corneal ulceration and, possibly, visual loss. The purpose of this study was to compare different nerve-grafting techniques to reanimate the paralyzed eyelid. Sixteen adult dogs (25 kg each) were allocated into four groups. Denervation of the left hemi-face was performed in all cases. One dog served as a control animal (group I). Group II dogs (n = 5) underwent end-to-side coaptation of the nerve graft to the intact palpebral branch and end-to-end coaptation to the denervated palpebral branch. Group III dogs (n = 5) underwent end-to-end coaptation of the nerve graft to the intact palpebral branch and end-to-end coaptation to the denervated palpebral branch. Group IV dogs (n = 5) underwent end-to-side coaptation of the nerve graft to the intact and denervated palpebral branches. The animals were monitored for 9 months after the surgical procedures, to allow adequate time for reinnervation. The dogs were postoperatively monitored with clinical observation, electrophysiologic testing, video motion analysis, and histologic assessments. Clinical observation and electrophysiologic testing demonstrated the production of an eye blink in the denervated hemi-face in all experimental groups. There was a trend toward increased speed of reinnervation for group III animals (end-to-end coaptations). It was concluded that end-to-side coaptation can produce a contralateral synchronous eye blink in a clinically relevant, large-animal model.

Sungpet, A., C. Suphachatwong, et al. (2003). "One-fascicle median nerve transfer to biceps muscle in C5 and C6 root avulsions of brachial plexus injury." Microsurgery 23(1): 10-3.
 Five male patients with avulsions of the C5 and C6 roots of the brachial plexus underwent transfer of one fascicle of the median nerve to the motor branch of the biceps muscle. The mean period of follow-up was 32 months. The average reinnervation time of the biceps was 3.4 months. Four patients achieved biceps strength of Medical Research Council (MRC) grade 4, and one patient had strength of the biceps of MRC grade 3. The mean period of time from surgery to MRC grade 3 was 9 months. At the last follow-up examination, grip strength, pinch strength, moving two-point discrimination, and strength of wrist volar flexion on the affected side were not worse than before the operation in any patient.

Sungpet, A., C. Suphachatwong, et al. (2003). "Transfer of one fascicle of ulnar nerve to functioning free gracilis muscle transplantation for elbow flexion." ANZ J Surg 73(3): 133-5.
 BACKGROUND: In brachial plexus injury, elbow flexion is the first priority in reconstruction. Neglected cases need functioning free muscle transplantation that requires the donor nerve to supply the transplanted muscle. The purpose of this study was to investigate the effects and results of transferring one fascicle of the ulnar nerve to the transplanted gracilis muscle. METHODS: One woman and two men with neglected avulsions of the C5,C6 roots of the brachial plexus underwent free gracilis muscle transfer for elbow flexion. One fascicle of the ulnar nerve was used as the donor nerve. RESULTS: The mean period of follow-up was 33.3 months. The average reinnervation time of gracilis muscle was 3.7 months. At the final examination, the mean strength of elbow flexion was 4.3 kgf. The grip strength, moving two-point discrimination and the strength of the wrist volar flexion on the affected side was not worse than before surgery in any patient at the last follow-up examination. CONCLUSIONS: A fascicle of the ulnar nerve can be one of the most effective options for functioning free muscle transplantation for elbow flexion.

Suuronen, E. J., M. Nakamura, et al. (2003). "Innervated human corneal equivalents as in vitro models for nerve-target cell interactions." Faseb J.
 A sensory nerve supply is crucial for optimal tissue function. However, the mechanisms for successful innervation and the signaling pathways between nerves and their target tissue are not fully understood. Engineered tissue substitutes can provide controllable environments in which to study tissue innervation. We have therefore engineered human corneal substitutes that promote nerve in-growth in a pattern similar to in vivo re-innervation. We demonstrate that these nerves (a) are morphologically equivalent to natural corneal nerves; (b) make appropriate contact with target cells; (c) can generate action potentials; (d) respond to chemical and physical stimuli; and (e) play an important role in the overall functioning of the bioengineered tissue. This model can be used for studying the more general topics of nerve ingrowth or regeneration and the interaction between nerves and their target cells and, more specifically, the role of nerves in corneal function. This model could also be used as an in vitro alternative to animals for safety and efficacy testing of chemicals and drugs.

Suzuki, Y., S. Nakagomi, et al. (2003). "Collapsin response mediator protein-2 accelerates axon regeneration of nerve-injured motor neurons of rat." J Neurochem 86(4): 1042-50.
 The rat collapsin response mediator protein-2 (CRMP-2) is a member of CRMP family (CRMP-1-5). The functional consequence of CRMP-2 during embryonic development, particularly in neurite elongation, is relatively understood; however, the role in nerve regeneration is unclear. Here we examined the role of CRMP-2 during nerve regeneration using rat hypoglossal nerve injury model. Among the members, CRMP-1, CRMP-2, CRMP-5 mRNA expressions increased after nerve injury, whereas CRMP-3 and CRMP-4 mRNA did not show any significant change. In the N1E-115 cells, CRMP-2 has the most potent neurite elongation activity among the CRMP family members. In dorsal root ganglion (DRG) organ culture, CRMP-2 overexpression by adenoviral vector demonstrated substantial neurite elongation. On the other hand, CRMP-2 (DeltaC381), which acts as a dominant negative form of CRMP-2, inhibited neurite formation. Collectively, it would be plausible that CRMP-2 has potent nerve regeneration activity after nerve injury. We therefore examined whether CRMP-2 overexpression in the injured hypoglossal motor neurons accelerates nerve regeneration. A retrograde-tracer, Fluoro-Gold (FG), was used to evaluate the number of reprojecting motor neurons after nerve injury. CRMP-2-overexpressing motor neurons demonstrated the accelerated reprojection. The present study suggests that CRMP-2 has potent neurite elongation activity in nerve regeneration in vivo.

Suzuki, M., S. Itoh, et al. (2003). "Tendon chitosan tubes covalently coupled with synthesized laminin peptides facilitate nerve regeneration in vivo." J Neurosci Res 72(5): 646-59.
 We have developed tendon chitosan tubes having the ability to bind peptides covalently, and the effectiveness of laminin peptides coupled to these tubular wall on nerve regeneration was examined in vivo. Bridge graft implantation (15 mm) into the sciatic nerve of SD rats was carried out using chitosan tubes having a triangular cross section containing either covalently bound intact laminin or the laminin peptides CDPGYIGSR or CSRARKQAASIKVAVSAD or being nontreated (N = 20 in each group). As a control, isografting (N = 5) was carried out. Three rats in each experimental group were sacrificed for histology observations after 1, 2, 4, 6, and 8 weeks. The total area of regenerating tissue in the tube and the length of the area where regenerating tissue attached to the inner surface of the tube were measured. In five rats from each experimental and control group, the latency quotient between the implanted and the nontreated site was determined 12 weeks after implantation. Furthermore, the percentage of myelinated axon area was measured at a 10-mm distance from the distal anastomosed site. Histological findings suggest that the immobilized laminin, confirmed by immunostaining as long as 12 weeks postoperatively, as well as laminin oligopeptides may effectively assist nerve tissue extension. According to statistical analysis of the percentage neural tissue found in relation to evoked action potentials, the sequential treatments with YIGSR first followed by IKVAV matched the effectiveness of intact laminin in enhancing nerve regeneration. However, when compared with that after isografting, the enhancement of regenerated axon growth was less sufficient.

Swietaszczyk, C., J. Maciaczyk, et al. (2003). "[In Process Citation]." Przegl Lek 60(5): 371-4.
 Although peripheral nerve regeneration is not a precise renewal of primary connections, it often leads to successful return of their function. Moreover, in some cases, an injured nerve is aimly stitched to another one. A good example of such a procedure is intercostal neurotization of avulsed brachial plexus. Despite the fact that neural centers are not primarily suitable for their new function, they often fulfill it successfully. This respecification of function requires some changes in the central nervous system called "plasticity". Discussed are the potential mechanisms controlling these changes. The role of hypothetic proteins reversely transported from the target organ to the neuron's body, proprioceptive fibers and volitional control is mentioned. The punctual understanding of these mechanisms may appear not only very scientifically wonderful but also useful in clinical practice.

Szabo, M., E. E. Salpeter, et al. (2003). "Transients in acetylcholine receptor site density and degradation during reinnervation of mouse sternomastoid muscle." J Neurochem 84(1): 180-8.
 The degradation rates of acetylcholine receptors (AchRs) were evaluated at the neuromuscular junction during and just after reinnervation of denervated muscles. When mouse sternomastoid muscles are denervated by multiple nerve crush, reinnervation begins 2-4 days later and is complete by day 7-9 after the last crush. In fully innervated muscles, the AChR degradation rate is stable and slow (t1/2 approximately 10 days), whereas after denervation the newly inserted receptors degrade rapidly (t1/2 approximately 1.2 days). The composite profile of degradation, which a mixture of the stable and the rapid receptors would give, is not observed during reinnervation. Instead, the receptors inserted between 2.5 and 7.5 days after the last crush all have an intermediate degradation rate of t1/2 approximately 3.7 days with standard error +/- 0.3 days. The total receptor site density at the endplate was evaluated during denervation and during reinnervation. As predicted theoretically, the site density increased substantially, but temporarily, after denervation. An analogous deleterious substantial decrease in density would be expected during reinnervation, without the intermediate receptor. This decrease is not observed, however, because of a large insertion rate at intermediate times (3000 +/- 700 receptor complexes per micro m2 per day). The endplate density of receptors thus remains relatively constant.

Tai, M. H., H. Cheng, et al. (2003). "Gene transfer of glial cell line-derived neurotrophic factor promotes functional recovery following spinal cord contusion." Exp Neurol 183(2): 508-15.
 Neuronal cell death and the failure of axonal regeneration cause a permanent functional deficit following spinal cord injury (SCI). Administration of recombinant glial cell line-derived neurotrophic factor (GDNF) has previously been reported to rescue neurons following severe SCI, resulting in improved hindlimb locomotion in rats. In this study, thus, GDNF gene therapy using an adenoviral vector (rAd-GDNF) was examined in rats following SCI induced by dropping the NYU weight-drop impactor from a height of 25 mm onto spinal segment T9-T10. To evaluate the efficacy of intraspinal injection of recombinant adenovirus into the injured spinal cord, we observed green fluorescent protein (GFP) gene transfer in the contused spinal cord. GFP was effectively expressed in the injured spinal cord, and the most prominently transduced cells were astrocytes. The expression of GDNF was detected only in rats receiving rAd-GDNF, not the controls, and remained detectable around the injured site for at least 8 days. Open-field locomotion analysis revealed that rats receiving rAd-GDNF exhibited improved locomotor function and hindlimb weight support compared to the control groups. Immunohistochemical examination for the neuronal marker, calcitonin gene-related peptide (CGRP), showed an increase in CGRP+ neuronal fibers in the injured spinal cord in rats receiving rAd-GDNF treatment. Collectively, the results suggest that adenoviral gene transfer of GDNF can preserve neuronal fibers and promote hindlimb locomotor recovery from spinal cord contusion. This research should provide information for developing a clinical strategy for GDNF gene therapy.

Taherzadeh, O., W. R. Otto, et al. (2003). "Influence of human skin injury on regeneration of sensory neurons." Cell Tissue Res 312(3): 275-80.
 The regeneration of sensory nerve fibres is regulated by trophic factors released from their target tissue, particularly the basal epidermis, and matrix molecules. Means to modulate this response may be useful for the treatment of neuromas and painful hypertrophic scars and of sensory deficits in skin grafts and flaps. We have developed an in vitro model of sensory neuron regeneration on human skin in order to study the mechanisms of sensory dysfunction in pathological conditions. Adult rat sensory neurons were co-cultured with unfixed cryosections of normal or injured (crushed) human skin for 72 h. Neurons were immunostained for growth-associated protein-43 and the neurite lengths of neuronal cell bodies situated in various skin regions were measured. Two-way analysis of variance was performed. Neurites of sensory cell bodies on epidermis of normal skin were the shortest, with a mean +/- SEM of 75+/-10 micrometer, whereas those of cells on the dermo-epidermal junction were the longest, with a mean +/- SEM of 231+/-18 micrometer. Neurons on the dermo-epidermal junction of injured skin had significantly longer neurites than those on the same region of normal skin (mean +/- SEM = 289+/-21 micrometer). Regeneration of sensory neurons may be influenced by extracellular matrix molecules, matrix-binding growth factors and trophic factors. Altered substrate or trophic factors in injured skin may explain the increase of neurite lengths. This in vitro model may be useful for studying the molecular mechanisms of sensory recovery and the development of neuropathic pain following peripheral nerve injury.

Takahashi, M., Y. Arai, et al. (2003). "Ependymal cell reactions in spinal cord segments after compression injury in adult rat." J Neuropathol Exp Neurol 62(2): 185-94.
 Recently, it has been suggested that neural stem cells and neural progenitor cells exist in the ependyma that forms the central canal of the spinal cord. In this study, we produced various degrees of thoracic cord injury in adult rats using an NYU-weight-drop device, assessed the degree of recovery of lower limb motor function based on a locomotor rating scale, and analyzed the kinetics of ependymal cell proliferation and differentiation by proliferating cell nuclear antigen (PCNA), nestin, glial fibrillary acidic protein (GFAP), or GAP-43 immunostaining. The results showed that the time course of the ependymal cell proliferation and differentiation reactions differed according to the severity of injury, and that the responses occurred not only in the neighborhood of the injury but in the entire spinal cord. An increase in the locomotor rating score was related to an increase in the number of PCNA-positive cells, and the differentiation of ependymal cells into reactive astrocytes was involved in injury repair. No apoptotic cells in the ependyma were detectable by the TUNEL method. These results indicate that the ependymal cells of the spinal central canal are themselves multipotent, can divide and proliferate according to the severity of injury, and differentiate into reactive astrocytes within the ependyma without undergoing apoptosis or cell death.

Takeda, M., M. Okochi, et al. (2003). "[Research strategy for understanding neuronal plasticity and degeneration]." Seishin Shinkeigaku Zasshi 105(1): 43-6.
 After the disclosure of human genomic information in 2003, the new era of post-genomic research will be in full bloom in the field of neuroscience, brain science as well as in neuropsychiatry. In this paper, the authors discussed possible strategies for understanding neuronal plasticity and neuronal degeneration in the post-genomic research paradigm. Five presentations are discussed in the following orders; 1) pharmacogenomics and epigenetics, 2) neuronal stem cell differentiation and transplantation into brain tissue, 3) Elucidation of the mechanism of neuro-protection and neurogenesis, 4) mechanisms of protein processing, especially importance of regulated intramembranous proteolysis, and 5) simulation analysis of signal transduction pathways. All of these newly developing strategies will be of increasing importance in the research in neuropsychiatry.

Tam, S. L. and T. Gordon (2003). "Neuromuscular activity impairs axonal sprouting in partially denervated muscles by inhibiting bridge formation of perisynaptic Schwann cells." J Neurobiol 57(2): 221-34.
 Following partial denervation of rat hindlimb muscle, terminal Schwann cells extend processes from denervated endplates to induce and guide sprouting from the remaining intact axons. Increased neuromuscular activity significantly reduces motor unit enlargement and sprouting during the acute phase of sprouting. These findings led to the hypothesis that increased neuromuscular activity perturbs formation of Schwann cell bridges and thereby reduces sprouting. Adult rat tibialis anterior (TA) muscles were extensively denervated by avulsion of L4 spinal root and were immediately subjected to normal caged activity or running exercise (8 h daily) for 3, 7, 14, 21, and 28 days. Combined silver/cholinesterase histochemical staining revealed that the progressive reinnervation of denervated endplates by sprouts over a 1 month period in the extensively partially denervated TA muscles was completely abolished by increased neuromuscular activity. Immunohistochemical staining and triple immunofluorescence revealed that the increased neuromuscular activity did not perturb the production of Schwann cell processes, but prevented bridging between Schwann cell processes at innervated and denervated endplates. Our findings suggest that failure of Schwann cell processes to bridge between endplates accounts, at least in part, for the inhibitory effect of increased neuromuscular activity on sprouting.

Tanabe, K., I. Bonilla, et al. (2003). "Fibroblast growth factor-inducible-14 is induced in axotomized neurons and promotes neurite outgrowth." J Neurosci 23(29): 9675-86.
 For successful nerve regeneration, a coordinated shift in gene expression pattern must occur in axotomized neurons. To identify genes participating in axonal regeneration, we characterized mRNA expression profiles in dorsal root ganglia (DRG) before and after sciatic nerve transection. Dozens of genes are differentially expressed after sciatic nerve injury by microarray analysis. Induction of SOX11, FLRT3, myosin-X, and fibroblast growth factor-inducible-14 (Fn14) mRNA in axotomized DRG neurons was verified by Northern analysis and in situ hybridization. The Fn14 gene encodes a tumor necrosis-like weak inducer of apoptosis (TWEAK) receptor and is dramatically induced in DRG neurons after nerve damage, despite low expression in developing DRG neurons. Fn14 expression in PC12 cells is also upregulated by nerve growth factor treatment. Overexpression of Fn14 promotes growth cone lamelipodial formation and increases neurite outgrowth in PC12 cells. These Fn14 effects are independent of the ligand, TWEAK. Fn14 colocalizes with the Rho family GTPases, Cdc42 and Rac1. Furthermore, Fn14 physically associates with Rac1 GTPase in immunoprecipitation studies. The neurite outgrowth-promoting effect of Fn14 is enhanced by Rac1 activation and suppressed by Rac1 inactivation. These findings suggest that Fn14 contributes to nerve regeneration via a Rac1 GTPase-dependent mechanism.

Tanaka, K., N. Fujita, et al. (2003). "Immunosuppressive (FK506) and non-immunosuppressive (GPI1046) immunophilin ligands activate neurotrophic factors in the mouse brain." Brain Res 970(1-2): 250-3.
 Based on the fact that several recent reports have indicated that non-immunosuppressive immunophilin ligands (IPLs) can activate neurite outgrowth or nerve regeneration, we investigated the neurotrophic factor-activating abilities of IPLs in vivo in order to clarify the molecular basis of neurotrophic-like activity. Both FK506 (an immunosuppressive IPL) and GPI1046 (a non-immunosuppressive IPL) significantly increased glial cell line-derived neurotrophic factor (GDNF) content in the substantia nigra. In addition, FK506 increased striatal brain-derived neurotrophic factor (BDNF) content significantly. Thus, our present results suggest that the molecular basis of IPL-induced neurotrophic-like activity may be dependent on GDNF and/or BDNF activation.

Tang, X., J. E. Davies, et al. (2003). "Changes in distribution, cell associations, and protein expression levels of NG2, neurocan, phosphacan, brevican, versican V2, and tenascin-C during acute to chronic maturation of spinal cord scar tissue." J Neurosci Res 71(3): 427-44.
 Previous studies have correlated the failure of axon regeneration after spinal cord injury with axons contacting scar tissue rich in chondroitin sulfate proteoglycans (CSPGs; Davies et al., 1999). In the present study, we have conducted immunohistochemical and quantitative Western blot analysis of five axon-growth-inhibitory CSPGs and tenascin-C within stab injuries of adult rat spinal cord at time points ranging from 24 hr to 6 months post injury. Quantitative Western blot analysis showed robust increases in neurocan, tenascin-C, and NG2 levels by 24 hr, suggesting that these molecules play a role in preventing axon regeneration across acutely forming scar tissue. Peak levels of 245/130 kD neurocan, NG2, and 250/200 kD tenascin-C were reached at 8 days, with maximum levels of phosphacan and 140/80 kD brevican attained later, at 1 month post injury. Versican V2 protein levels, however, displayed an opposite trend, dropping below unlesioned spinal cord values at all time points studied. Confocal microscopy at 8 days post injury revealed heightened immunoreactivity for phosphacan, NG2, and tenascin-C, particularly within fibronectin(+) scar tissue at lesion centers. In contrast, neurocan was displayed within lesion margins on the processes of stellate NG2(+) cells and, to a much lesser extent, by astrocytes. At 6 months post injury, 130 kD neurocan, brevican, and NG2 levels within chronic scar tissue remained significantly above control. Our results show novel expression patterns and cell associations of inhibitory CSPGs and tenascin-C that have important implications for axon regeneration across acute and chronic spinal cord scar tissue.

Tang, B. L. (2003). "Inhibitors of neuronal regeneration: mediators and signaling mechanisms." Neurochem Int 42(3): 189-203.
 Neuritogenesis and its inhibition are opposite and balancing processes during development as well as pathological states of adult neuron. In particular, the inability of adult central nervous system (CNS) neurons to regenerate upon injury has been attributed to both a lack of neuritogenic ability and the presence of neuronal growth inhibitors in the CNS environment. I review here recent progress in our understanding of neuritogenic inhibitors, with particular emphasis on those with a role in the inhibition of neuronal regeneration in the CNS, their signaling cascades and signal mediators. Neurotrophines acting through the tropomyosin-related kinase (Trk) family and p75 receptors promote neuritogenesis, which appears to require sustained activation of the mitogen activated protein (MAP) kinase pathway, and/or the activation of phosphotidylinositol 3-kinase (PI3 kinase). During development, a plethora of guidance factors and their receptors navigate the growing axon. However, much remained to be learned about the signaling receptors and pathways that mediate the activity of inhibitors of CNS regeneration. There is growing evidence that neuronal guidance molecules, particularly semaphorins, may also have a role as inhibitors of CNS regeneration. Although direct links have not yet been established in many cases, signals from these agents may ultimately converge upon the modulators and effectors of the Rho-family GTPases. Rho-family GTPases and their effectors modulate the activities of actin modifying molecules such as cofilin and profilin, resulting in cytoskeletal changes associated with growth cone extension or retraction.

Tartakovskaia, O. S., S. L. Borisenko, et al. (2003). "[Age factor in eye regeneration of the gastropod mollusk Achatina fulica]." Izv Akad Nauk Ser Biol(3): 285-92.
 The dependence of the ability to regenerate the eye on the age of experimental animals was studied in the snail Achatina fulica. The degree of regeneration was estimated by light-microscopic and electrophysiological methods and by analyzing the motor response to visual stimuli. In older age groups, the number of regenerated eye-bearing tentacles decreased, whereas the period of regeneration increased. The regenerated eyes of the snails operated at the age of more than two months remained smaller than normal eyes even after six months. Regeneration of the distal part of the optic nerve was observed, and the regenerated eyes recovered the ability to respond to stimulation by light. In the electroretinogram, the responses of the regenerated eye, compared to the control, were characterised by a lower amplitude and longer repolarization and refractory periods. Manifestations of the motor response to visual stimuli in the young snails with regenerating eyes could be regarded as evidence for the recovery of connection between the organ of sight and the central ganglia.

Tassava, R. A. and C. L. Olsen-Winner (2003). "Responses to amputation of denervated ambystoma limbs containing aneurogenic limb grafts." J Exp Zoolog Part A Comp Exp Biol 297(1): 64-79.
 The developing neural tubes and associated neural crest cells were removed from stage 30 Ambystoma maculatum embryos to obtain larvae with aneurogenic forelimbs. Forelimbs were allowed to develop to late 3 digit or early 4 digit stages. Limbs amputated through the mid radius-ulna regenerated typically in the aneurogenic condition. Experiments were designed to test whether grafts of aneurogenic limb tissues would rescue denervated host limb stumps into a regeneration response. In Experiment 1, aneurogenic limbs were removed at the body wall and grafted under the dorsal skin of the distal end of amputated forelimbs of control, normally innervated limbs of locally collected Ambystoma maculatum or axolotl (Ambystoma mexicanum) larvae. In Experiment 1, at the time of grafting or 1, 2, 3, 4, 5, 7, or 8 days after grafting, aneurogenic limbs were amputated level with the original host stump. At 7 and 8 days, this amputation included removing the host blastema adjacent to the graft. The host limb was denervated either one day after grafting or on the day of graft amputation. These chimeric limbs only infrequently exhibited delayed blastema formation. Thus, not only did the graft not rescue the host, denervated limb, but the aneurogenic limb tissues themselves could not mount a regeneration response. In Experiment 2, the grafted aneurogenic limb was amputated through its mid-stylopodium at 3, 4, 5, 7, or 8 days after grafting. By 7 and 8 days after grafting, the host limb stump exhibited blastema formation even with the graft extending out from under the dorsal skin. The host limb was denervated at the time of graft amputation. When graft limbs of Experiment 2 were amputated and host limbs were denervated on days 3, 4, or 5, host regeneration did not progress and graft regeneration did not occur. But, when graft limbs were amputated on days 7 or 8 with concomitant denervation of the host limb, regeneration of the host continued and graft regeneration occurred. Thus, regeneration of the graft was correlated with acquisition of nerve-independence by the host limb blastema. In Experiment 3, aneurogenic limbs were grafted with minimal injury to the dorsal skin of neurogenic hosts. When neurogenic host limbs were denervated and the aneurogenic limbs were amputated through the radius/ulna, regeneration of the aneurogenic limb occurred if the neurogenic limb host was not amputated, but did not occur if the neurogenic limb host was amputated. Results of Experiment 3 indicate that the inhibition of aneurogenic graft limb regeneration on a denervated host limb is correlated with substantial injury to the host limb. In Experiment 4, aneurogenic forelimbs were amputated through the mid-radius ulna and pieces of either peripheral nerve, muscle, blood vessel, or cartilage were grafted into the distal limb stump or under the body skin immediately adjacent to the limb at the body wall. In most cases, peripheral nerve inhibited regeneration, blood vessel tissue sometimes inhibited, but other tissues had no effect on regeneration. Taken together, the results suggest: (1) Aneurogenic limb tissues do not produce the neurotrophic factor and do not need it for regeneration, and (2) there is a regeneration-inhibiting factor produced by the nerve-dependent limb stump/blastema after denervation that prevents regeneration of aneurogenic limbs.

Tawk, M. and S. Vriz (2003). "[Regeneration of vertebrate appendage: an old experimental model to study stem cells in the adult]." Med Sci (Paris) 19(4): 465-71.
 The application of stem cell therapy to cure degenerative diseases offers immense possibilities, but the research in this field is the subject of ethical debates raised by the question of destructive research on early human embryos. Stem cells taken in the adult constitute an alternative to human embryonic stem cells, but our knowledge on totipotent or pluripotent cells is currently insufficient. Furthermore, many questions must be solved before selection and differentiation of these cells in a given cellular type can be controlled on a routine basis. What are the molecular characteristics of an adult stem cell? What are the mechanisms involved in cell reprogramming? Which signals control stem cell replication and differentiation? Basic research activities must be carried out in order to clarify all these points. In this context, the regeneration of vertebrate appendages provides a model for this type of research. The regeneration process is defined by both the morphological and functional reconstruction of a part of a living organism, which has previously been destroyed. But why are some vertebrates able to regenerate complex structures and others apparently not? Among most vertebrates, the capacity to regenerate is limited to some tissues. It is however possible to observe the regeneration of appendages (limb, tail, fin, jaw, etc.) among several amphibians and fish. This regeneration leads to re-forming of the amputated part with a complete restoration of its shape, segmentation and function. Why is the amputation of limbs not followed by regeneration in mammals and birds: absence of stem cells, absence of recruitment signals for these cells, or absence of signal receptivity? This review constitutes a report on the current understanding of the basis of on regeneration of legs in tetrapods and of fins in fish with an emphasis in the role of the nervous system in this process.

Teive, H. A., A. R. Troiano, et al. (2003). "Botulinum toxin for treatment of Frey's syndrome: report of two cases." Arq Neuropsiquiatr 61(2A): 256-8.
 Frey's syndrome is a phenomenon of hemifacial flushing and sweating after gustatory stimulus, usually secondary to surgical trauma over the parotid gland, although other injury mechanisms may be seen. It is accepted as a result of aberrant regeneration of facial autonomic nerve fibers. Treatment evolved from ineffective medical and surgical approaches to botulinum toxin. We evaluate the effectiveness and safety of botulinum toxin in the treatment of this complication in two patients.

Temple, M. D., P. F. Worley, et al. (2003). "Visualizing changes in circuit activity resulting from denervation and reinnervation using immediate early gene expression." J Neurosci 23(7): 2779-88.
 We describe a novel strategy to evaluate circuit function after brain injury that takes advantage of experience-dependent immediate early gene (IEG) expression. When normal rats undergo training or are exposed to a novel environment, there is a strong induction of IEG expression in forebrain regions, including the hippocampus. This gene induction identifies the neurons that are engaged during the experience. Here, we demonstrate that experience-dependent IEG induction is diminished after brain injury in young adult rats (120-200 gm), specifically after unilateral lesions of the entorhinal cortex (EC), and then recovers with a time course consistent with reinnervation. In situ hybridization techniques were used to assess the expression of the activity-regulated cytoskeleton-associated protein Arc at various times after the lesion (4, 8, 12, 16, or 30 d). One group of rats was allowed to explore a complex novel environment for 1 hr; control operated animals remained in their home cage. In unoperated animals, exposure to the novel environment induced Arc mRNA levels in most pyramidal neurons in CA1, in many pyramidal neurons in CA3, and in a small number of dentate granule cells. This characteristic pattern of induction was absent at early time points after unilateral EC lesions (4 and 8 d) but recovered progressively at later time points. The recovery of Arc expression occurred with approximately the same time course as the reinnervation of the dentate gyrus as a result of postlesion sprouting. These results document a novel approach for quantitatively assessing activity-regulated gene expression in polysynaptic circuits after trauma.

Ter Laak, M. P., J. H. Brakkee, et al. (2003). "The potent melanocortin receptor agonist melanotan-II promotes peripheral nerve regeneration and has neuroprotective properties in the rat." Eur J Pharmacol 462(1-3): 179-83.
 The neurotrophic and neuroprotective potential of the alpha-melanocyte-stimulating hormone (alpha-MSH) analog cyclo-[Ac-Nle(4),Asp(5),D-Phe(7),Lys(10)]alpha-MSH-(4-10) amide (melanotan-II), a potent melanocortin receptor agonist, was investigated. The sciatic nerve crush model was used as a paradigm to investigate the neurotrophic properties of melanotan-II. Melanotan-II significantly enhanced the recovery of sensory function following a crush lesion of the sciatic nerve in the rat at a dose of 20 microg kg(-1) per 48 h, s.c., but not at a dose of 2 or 50 microg kg(-1). In addition, we observed that melanotan-II also possesses neuroprotective properties, as it partially protected the nerve from a toxic neuropathy induced by cisplatin. Thus, the present data for the first time demonstrate the effectiveness of the potent alpha-MSH analog melanotan-II in nerve regeneration and neuroprotection.

Tervo, T. and J. Moilanen (2003). "In vivo confocal microscopy for evaluation of wound healing following corneal refractive surgery." Prog Retin Eye Res 22(3): 339-58.
 Understanding of corneal wound healing plays an important role, not only in management of corneal infections, but especially in refractive surgery. A better control of wound healing mechanisms might improve the results of such resculpturing techniques and help to avoid complications arising from these procedures. While studies have been focused in different aspects of corneal wound healing, our knowledge has increased greatly during the last years. Many problems associated with corneal healing also contribute to clinical pathology following corneal surgery. Understanding of such conditions has been augmented by the continuously developing corneal imaging techniques. We have used in vivo confocal microscopy (IVCM) for assessing corneas subjected to refractive surgery as well as corneas with common complications resulting from such procedures. IVCM has become a powerful tool for examining corneal cells, nerves, inflammations and infections. It allows information to be acquired repeatedly and at subbiomicroscopic levels that earlier had been obtainable only by invasive microscopic methods. Pre-examining corneas preoperatively by IVCM in order to reveal diseases or conditions in which elective refractive surgical procedures should not be undertaken or to select the ideal operation technique may help to avoid complications in the future. Measurement of the thickness of corneal sublayers or estimation of the thickness of a laser in situ keratomileusis flap or wound bed are other applications in which confocal microscopy may be valuable. In this article we attempt to describe the in vivo confocal findings of common refractive procedures and their complications, and discuss their biology in light of the existing knowledge on wound healing phenomena.

Thomas, D. R. (2003). "The promise of topical growth factors in healing pressure ulcers." Ann Intern Med 139(8): 694-5.
 
Thomas, C. K., S. Sesodia, et al. (2003). "Properties of medial gastrocnemius motor units and muscle fibers reinnervated by embryonic ventral spinal cord cells." Exp Neurol 180(1): 25-31.
 Severe muscle atrophy occurs after complete denervation. Here, Embryonic Day 14-15 ventral spinal cord cells were transplanted into the distal tibial nerve stump of adult female Fischer rats to provide a source of neurons for muscle reinnervation. Our aim was to characterize the properties of the reinnervated motor units and muscle fibers. Some reinnervated motor units contracted spontaneously. Electrical stimulation of the transplants at increasing intensity produced an average (+/- SE) of 7 +/- 1 electromyographic and force steps. Each signal increment represented the excitation of another motor unit. These reinnervated units exerted an average force of 12.0 +/- 1.5 mN, strength similar to that of control fatigue-resistant units. Repeated transplant stimulation depleted 17% of the muscle fibers of glycogen, an indication of some functional reinnervation. Reinnervated (glycogen-depleted), denervated (no cells transplanted), and control fibers were of histochemical type I, IIA, or IIB. Fibers of the same type were grouped after reinnervation. The proportion of fiber types also changed. Reinnervated fibers were primarily type IIA, whereas most fibers in denervated and control muscles were type IIB. Reinnervated fibers of each type had significantly larger cross-sectional areas than the corresponding fiber types in denervated muscles. These data suggest that neurons with different properties can reside in the unusual environment of the adult rat peripheral nerve, make functional connections with muscle, specify muscle fiber type, and reduce the amount that each type atrophies.

Thompson, K. M., N. Uetani, et al. (2003). "Receptor protein tyrosine phosphatase sigma inhibits axonal regeneration and the rate of axon extension." Mol Cell Neurosci 23(4): 681-92.
 Transgenic mice lacking receptor protein tyrosine phophatase-sigma (RPTPsigma), a type IIa receptor protein tyrosine phosphatase, exhibit severe neural developmental deficits. Continued expression of RPTPsigma in the adult suggests that it plays a functional role in the mature nervous system. To determine if RPTPsigma might influence axonal regeneration, the time course of regeneration following facial nerve crush in wild-type and RPTPsigma (-/-) mice was compared. Mice lacking RPTPsigma exhibited an accelerated rate of functional recovery. Immunocytochemical examination of wild-type neurons in cell culture showed RPTPsigma protein in the growth cone. To determine if RPTPsigma affects the ability of a neuron to extend an axon, the rate of axon growth in neuronal cultures derived from wild-type and RPTPsigma (-/-) embryonic mice was compared. RPTPsigma did not affect the rate of axon initiation, but the rate of axon extension is enhanced in neurons obtained from RPTPsigma (-/-) mice. These findings indicate that RPTPsigma slows axon growth via a mechanism intrinsic to the neuron and identify a role for RPTPsigma regulating axonal regeneration by motoneurons.

Timmer, M., S. Robben, et al. (2003). "Axonal regeneration across long gaps in silicone chambers filled with Schwann cells overexpressing high molecular weight FGF-2." Cell Transplant 12(3): 265-77.
 Basic fibroblast growth factor (FGF-2) has been shown to enhance the survival and neurite extension of various types of neurons including spinal ganglion neurons. In addition, endogenous FGF-2 and FGF receptors are upregulated following peripheral nerve lesion in ganglia and at the lesion site. FGF-2 protein is expressed in different isoforms (18 kDa, 21 kDa, 23 kDa) and differentially regulated after nerve injury. In the rat we analyzed the regenerative capacity of the high molecular weight (HMW) FGF-2 isoforms (21/23 kDa) to support the regeneration of the axotomized adult sciatic nerve across long gaps. The nerve stumps were inserted into the opposite ends of a silicone chamber resulting in an interstump gap of 15 mm. Silicone tubes were filled with Matrigel or a mixture of Schwann cells (SC) and Matrigel. SC were prepared from newborn rats and transfected to overexpress HMW FGF-2. Four weeks after the operation procedure, channels were analyzed with regard to tissue cables bridging both nerve stumps and myelinated axons distal to the original proximal nerve stump. Peripheral nerves interposed with HMW Schwann cells displayed significantly enhanced nerve regeneration, with the greatest number of tissue cables containing myelinated axons and the highest number of myelinated axons. These results suggest that a cellular substrate together with a source of a trophic factor could be a promising tool to promote nerve regeneration and, therefore, become useful also for a clinical approach to repair long gaps.

Tonchev, A. B., T. Yamashima, et al. (2003). "Proliferation of neural and neuronal progenitors after global brain ischemia in young adult macaque monkeys." Mol Cell Neurosci 23(2): 292-301.
 To investigate the effect of global cerebral ischemia on brain cell proliferation in young adult macaques, we infused 5-bromo-2'-deoxyuridine (BrdU), a DNA replication indicator, into monkeys subjected to ischemia or sham-operated. Subsequent quantification by BrdU immunohistochemistry revealed a significant postischemic increase in the number of BrdU-labeled cells in the hippocampal dentate gyrus, subventricular zone of the temporal horn of the lateral ventricle, and temporal neocortex. In all animals, 20-40% of the newly generated cells in the dentate gyrus and subventricular zone expressed the neural progenitor cell markers Musashi1 or Nestin. A few BrdU-positive cells in postischemic monkeys were double-stained for markers of neuronal progenitors (class III beta-tubulin, TUC4, doublecortin, or Hu), neurons (NeuN), or glia (S100beta or GFAP). Our results suggest that ischemia activates endogenous neuronal and glial precursors residing in diverse locations of the adult primate central nervous system.

Tonchev, A. B., T. Yamashima, et al. (2003). "Differential proliferative response in the postischemic hippocampus, temporal cortex, and olfactory bulb of young adult macaque monkeys." Glia 42(3): 209-24.
 We investigated the fate of proliferating cells in the adult monkey brain after global ischemia. We used the thymidine analogue bromodeoxyuridine (BrdU) to label S-phase cells and their progeny in Japanese macaques subjected to global cerebral ischemia for 20 min or to a sham operation. Subsequently, newly generated cells were identified by BrdU immunohistochemistry, and their immunophenotype was determined quantitatively, using specific markers. The ischemic insult significantly increased the number of proliferating cells in the hippocampus and temporal neocortex, where the majority BrdU-labeled cells expressed markers for microglia (Iba1, CD68, and Ham56) or astrocytes (S-100beta and glial fibrillary acidic protein [GFAP]). In contrast, the proliferation level in the parahippocampal region remained unchanged. This discrepancy prompted us to investigate the postischemic response in the olfactory bulb, a well-known site of adult cell generation that is anatomically distant from the above-mentioned regions but that is also subjected to the global ischemic insult. The olfactory bulb contained clusters of proliferating cells expressing markers for neural (Musashi1 and Nestin) and/or neuronal (class III beta-tubulin) progenitors; these were immunophenotypically distinct from other cell types. Their number and distribution were unaltered by ischemia. Our results demonstrate that cell proliferation and differentiation in the adult macaque brain and olfactory bulb are differentially affected by a common insult.

Tonn, J. C. and R. Goldbrunner (2003). "Mechanisms of glioma cell invasion." Acta Neurochir Suppl 88: 163-7.
 Invasive growth is one of the characteristics of gliomas--local infiltration into the surrounding nerve tissue decisively restricts all treatment strategies. Particularly the merit of all local treatment modalities is queried. The question whether a glioma represents a diffuse disease of the CNS or a local disturbance with unrestrained expansion tendency is still at issue. Understanding of the invasion mechanisms is of importance inasmuch as biologically reasonable and effective strategies of limiting and suppressing glioma invasion can only hence be derived. The affinity of glioma cells towards certain structures of the extracellular matrix as well as taking advantage of tumour vascularisation with regard to extension play a decisive role. Still not fully understood are tumour host interactions. Future thinking will have to take into account these interactions as well as evidence to be derived from development neurobiology and regeneration capacity of the CNS. The present review is meant to give a short overview and disclose many questions.

Toti, P., M. Villanova, et al. (2003). "Nerve growth factor expression in human dystrophic muscles." Muscle Nerve 27(3): 370-3.
 Nerve growth factor (NGF) is a neurotrophin that is expressed during muscle development and is also capable of favoring muscle regeneration in experimental studies. The presence of NGF in muscular dystrophies, such as Duchenne and Becker muscular dystrophies, has never been fully explored. By means of immunohistochemistry, we show that regenerating muscle fibers from such patients consistently express NGF, as do myofibroblasts and mast cells. By contrast, rest fibers from dystrophic patients, as well as muscle fibers from healthy, control patients and even regenerative muscle fibers in polymyositis do not show NGF immunoreactivity. The paracrine effect of NGF on muscle regeneration, as well as its chemoattractant capacities for mast cells, may contribute to explaining why regenerating fibers most frequently occur in clusters and why mast cells are more numerous in dystrophic muscles. Moreover, being a mediator of wound healing and tissue fibrosis, NGF may contribute to long-term muscle regeneration impairment by tissue fibrosis in the muscular dystrophies.

Toyoda, H., K. Ohno, et al. (2003). "Induction of NMDA and GABAA receptor-mediated Ca2+ oscillations with KCC2 mRNA downregulation in injured facial motoneurons." J Neurophysiol 89(3): 1353-62.
 To clarify the changes that occur in gamma-aminobutyric acid type A (GABA(A)) receptor-mediated effects and contribute to alterations in the network activities after neuronal injury, we studied intracellular Ca(2+) concentration ([Ca(2+)](i)) dynamics in a rat facial-nerve-transection model. In facial motoneurons, an elevation of the resting [Ca(2+)](i), GABA-mediated [Ca(2+)](i) transients, enhancement of the glutamate-evoked [Ca(2+)](i) increases, and spontaneous [Ca(2+)](i) oscillations were induced by axotomy. All these axotomy-induced modifications were abolished by the GABA(A)-receptor antagonist bicuculline and N-methyl-d-aspartate (NMDA)-receptor antagonist d(-)-2-amino-5-phosphonopentanoic acid. A downregulation of K(+)-Cl(-) cotransporter (KCC2) mRNA, an increase in intracellular Cl(-) concentration ([Cl(-)](i)), and transformation of GABAergic hyperpolarization to depolarization were also induced by axotomy. We suggest that in axotomized neurons KCC2 downregulation impairs Cl(-) homeostasis and makes GABA act depolarizing, resulting in endogenous GABA inducing [Ca(2+)](i) oscillations via facilitation of NMDA-receptor activation. Such GABA(A)-receptor-mediated [Ca(2+)](i) oscillations may play a role in neural survival and regeneration.

Tripanichkul, W., D. Stanic, et al. (2003). "D2 Dopamine receptor blockade results in sprouting of DA axons in the intact animal but prevents sprouting following nigral lesions." Eur J Neurosci 17(5): 1033-45.
 Recently it was demonstrated that sprouting of dopaminergic neurons and a microglial and astrocyte response follows both partial lesions of the substantia nigra pars compacta and blockade of the D2 dopamine receptor. We therefore studied the effects of the combination of these two treatments (lesioning and D2 dopamine receptor blockade). Haloperidol administration caused a 57% increase in dopaminergic terminal tree size (measured as terminal density per substantia nigra pars compacta neuron) and an increase of glia in the striatum. Following small to medium nigral lesions (less than 60%), terminal tree size increased by 51% on average and returned density of dopaminergic terminals to normal. In contrast, administration of haloperidol for 16 weeks following lesioning resulted in reduced dopaminergic terminal density and terminal tree size (13%), consistent with absent or impaired sprouting. Glial cell numbers increased but were less than with lesions alone. When haloperidol was administered after the striatum had been reinnervated through sprouting (16-32 weeks after lesioning), terminal tree size increased up to 150%, similar to the effect of haloperidol in normal animals. By examining the effect of administering haloperidol at varying times following a lesion, we concluded that a switch in the effect of D2 dopamine receptor blockade occurred after dopaminergic synapses began to form in the striatum. We postulate that when synapses are present, D2 dopamine receptor blockade results in increased terminal density, whereas prior to synapse formation D2 dopamine receptor blockade causes attenuation of a sprouting response. We speculate that D2 dopamine receptors located on growth cones 'push' neurites toward their targets, and blockade of these receptors could lead to attenuation of sprouting.

Tseng, C. Y., G. Hu, et al. (2003). "Histologic analysis of Schwann cell migration and peripheral nerve regeneration in the autogenous venous nerve conduit (AVNC)." J Reconstr Microsurg 19(5): 331-40.
 Over the last two decades, the autogenous venous nerve conduit (AVNC) has been established as an effective treatment modality for the repair of nerve gaps less than 3 cm. In this study, the spatial-temporal progression of Schwann-cell migration and peripheral-nerve regeneration across a 10-mm gap bridged by a venous conduit was examined, using immunoctyochemical techniques. Histologic analysis revealed that the process of nerve regeneration through an AVNC occurs in four phases: the hematoma phase, cellular migration phase, axonal advancement phase, and myelination and maturation phase. The authors found that: 1) the lumen of the vein conduit remains patent throughout the process of nerve regeneration; 2) Schwann cells migrate into the vital space of the vessel lumen from the proximal and distal nerve stumps; 3) axonal growth into the conduit lags behind Schwann-cell migration; 4) Schwann cells migrate to the regenerating axons to form mature nodes of Ranvier when the distal stump is present; and 5) mechanical injury alone is sufficient to induce axonal outgrowth from the proximal nerve stump.

Tsukatani, T., H. L. Fillmore, et al. (2003). "Matrix metalloproteinase expression in the olfactory epithelium." Neuroreport 14(8): 1135-40.
 The olfactory epithelium contains neuronal progenitor cells capable of continuous neurogenesis and is a unique model for studying neural degeneration, regeneration, axon outgrowth and recovery from injury. Matrix metalloproteinases (MMPs), and tissue inhibitors of metalloproteinases (TIMPs), have been implicated in cell turnover, development, migration, and metastatic processes. We used Western blot and immunohistochemistry to determine whether MMP-2 and associated proteins TIMP-2 and membrane type 1 matrix metalloproteinase (MT1-MMP) are present in the olfactory epithelium of mice. We found MMP-2 expression localized to the olfactory basal cells and immature neurons. After injury-induced neural degeneration, MMP-2 and MT1-MMP levels decreased while TIMP-2 levels increased. However, following 35 days of neurogenesis and cell replacement TIMP-2 and MT1-MMP returned to control levels. The results show a correlation between MMP and TIMP levels and the stages of neural degeneration, regeneration and recovery of the olfactory epithelium following injury.

Turbes, C. C. (2003). "Dorsal root implant on lesioned spinal cord morphologic findings of regeneration of synapses in the mammalian spinal cord--repair and recovery." Biomed Sci Instrum 39: 289-99.
 Earlier work concerning regeneration of synaptic connection had been studied primarily in amphibia. Sperry and Miner and Stevens showed that functional regeneration of synapses followed sectioning and anastomosis of the central process of the dorsal root of one side to the proximal stump of the dorsal root of the opposite. A number of studies have shown that the dorsal roots and dorsal columns of mammals have adequate regenerative capacities. There is no functional or morphologic evidence for reestablishment of synaptic connections reported in the mammalian studies. A number of authors have reported that regenerating dorsal root fibers are confronted with a barrier at the neutilemmal-glial junctions. Previous studies have shown that peripheral nerve fibers inserted into the spinal cord grow profusely. Theorizing that regenerating dorsal root fibers would grow and reestablish synaptic connection more readily, it was decided to insert the neurilemmal portion- including Schwann cells- of the dorsal roots into the spinal cord to the depths of the gray columns.

Tyler, D. J. and D. M. Durand (2003). "Chronic response of the rat sciatic nerve to the flat interface nerve electrode." Ann Biomed Eng 31(6): 633-42.
 The chronic effects of a reshaping nerve electrode, the flat interface nerve electrode (FINE), on sciatic nerve physiology, histology, and blood-nerve barrier (BNB) are presented. The FINE electrode applies a small force to a nerve to reshape the nerve and fascicles into elongated ovals. This increases the interface between the nerve and electrode for selective stimulation and recording of peripheral nerve activity. The hypothesis of this study is that a small force applied noncircumferentially to a nerve can chronically reshape the nerve without effecting nerve physiology, histology, or the blood-nerve barrier permeability. Three FINE electrode designs were implanted on rat sciatic nerves to examine the nerve's response to small, moderate, and high reshaping forces. The chronic reshaping, physiology, and histology of the nerve were examined at 1, 7, and 28 days postimplant. All FINEs significantly reshape both the nerve and the fascicles compared to controls. FINEs that applied high forces caused a neurapraxia type injury characterized by changes in the animal's footprint, nerve histology, and the BNB permeability. The physiological changes were greatest at 7 days and fully recover to normal by 14 days postimplant. The moderate force FINE did not result in changes in the footprint or BNB permeability. Only a minor decrease in axon density without accompanying evidence of axon demyelination or regeneration was observe for the moderate force. The small force FINE does not cause any change in nerve physiology, histology, or BNB permeability compared to the sham treatment. An electrode that applies a small force that results in an estimated intrafascicular pressure of less than 30 mm Hg can reshape the nerve without significant changes in the nerve physiology or histology. These results support the conclusion that a small force chronically applied to the nerve reshapes the nerve without injury.

Uchida, N., M. Kanazawa, et al. (2003). "Expression of BDNF and TrkB in mouse taste buds after denervation and in circumvallate papillae during development." Arch Histol Cytol 66(1): 17-25.
 BDNF (brain-derived neurotrophic factor) is a member of the neurotrophin family which affects the proliferation and survival of neurons. Using an immunocytochemical method, we examined the expression of BDNF and its receptor, TrkB, in the taste bud cells of the circumvallate papillae of normal mice and of mice after transection of the glossopharyngeal nerves. We additionally observed the expression of BDNF and TrkB in the developing circumvallate papillae of late prenatal and early postnatal mice. In normal untreated mice, BDNF was expressed in most of the taste bud cells; TrkB was detected in the plasma membrane of taste bud cells and in the nerve fibers. Double-labeling studies showed that BDNF and NCAM (neural cell adhesion molecule) or TrkB and NCAM colocalized in some of the taste bud cells, but that most taste bud cells were immunopositive for only BDNF or TrkB. NCAM-immunoreactive cells are known to be type-III cells, which have afferent synaptic contacts with the nerve terminals. Five days after denervation, the number of taste buds and nerve fibers markedly decreased; however, the remaining taste bud cells still expressed BDNF and TrkB. By 10 days after denervation, most of the taste buds had disappeared, and there were a few TrkB-immunoreactive nerve fibers in the connective tissue core. By 4 weeks after denervation, numerous TrkB-immunoreactive nerve fibers had invaded the papillae, and a few taste buds expressing BDNF and TrkB had regenerated. At E (embryonic day) 15 during development, the circumvallate papillae appeared, and then TrkB-immunoreactive nerve fibers entered the connective tissue core, and some of these fibers further invaded among the dorsal epithelial cells of the papillae. TrkB-immunoreactive oval-shaped cells were occasionally found in the dorsal epithelium. Such TrkB-immunoreactive nerve fibers and cells were also observed at E16-18. However, BDNF was not expressed in the papillae through the late prenatal days of E15 to E18. At P (postnatal day) 0, a cluster of BDNF-and TrkB-immunoreactive cells appeared in the dorsal epithelium of the papillae, and was presumed to be primitive taste buds. We conclude that TrkB-immunoreactive nerve fibers are necessary for papillary and taste bud formation during development and for the regeneration of taste buds after denervation. BDNF in the taste bud cells may act as a neurotrophic factor for innervating sensory neurons--through TrkB receptors of the axons of those neurons, and also may exert autocrine and paracrine trophic actions on neighboring taste bud cells by binding to their TrkB receptors.

Udina, E., D. Ceballos, et al. (2003). "FK506 enhances reinnervation by regeneration and by collateral sprouting of peripheral nerve fibers." Exp Neurol 183(1): 220-31.
 We examined the effects of FK506 administration on the degree of target reinnervation by regenerating axons (following sciatic nerve crush) and by collateral sprouts of the intact saphenous nerve (after sciatic nerve resection) in the mouse. FK506-treated animals received either 0.2 or 5 mg/kg/day, dosages previously found to maximally increase the rate of axonal regeneration in the mouse. Functional reinnervation of motor, sensory, and sweating activities was assessed by noninvasive methods in the hind paw over a 1-month period following lesion. Morphometric analysis of the regenerated nerves and immunohistochemical labeling of the paw pads were performed at the end of follow-up. In the sciatic nerve crush model, FK506 administration shortened the time until target reinnervation and increased the degree of functional and morphological reinnervation achieved. The recovery achieved by regeneration was greater overall with the 5 mg/kg dose than with the dose of 0.2 mg/kg of FK506. In the collateral sprouting model, reinnervation by nociceptive and sudomotor axons was enhanced by FK506. Here, the field expansion followed a faster course between 4 and 14 days in FK506-treated animals. In regard to dose, while collateral sprouting of nociceptive axons was similarly increased at both dosages (0.2 and 5 mg/kg), sprouting of sympathetic axons was more extensive at the high dose. This suggests that the efficacy of FK506 varies between subtypes of neurons. Taken together, our findings indicate that, in addition to an effect on rate of axonal elongation, FK506 improves functional recovery of denervated targets by increasing both regenerative and collateral reinnervation.

Udina, E., J. Voda, et al. (2003). "Comparative dose-dependence study of FK506 on transected mouse sciatic nerve repaired by allograft or xenograft." J Peripher Nerv Syst 8(3): 145-54.
 We evaluated the effects of FK506, at doses of 0.2, 2, and 5 mg/kg/day, on the response to nerve grafts implanted in outbred mice. A 6 mm long segment of the sciatic nerve was transected and repaired by autograft (the same segment resected), allograft (from another mouse), or xenograft (from a rat nerve). The regenerating nerves were harvested after 3 weeks and studied under light and electron microscope. Allografts of animals treated with the 5 mg/kg/day dose of FK506 appeared similar to those from autografts, demonstrating an equivalent number of myelinated fibers. In mice treated with the 2 mg/kg/day dose, regeneration was slightly hindered, as indicated by the reduced number of myelinated fibers. In contrast, in mice given a 0.2 mg/kg/day dose of FK506, allografts were not different from untreated allografts; both groups showed a marked rejection response with only few unmyelinated axons and no myelinated fibers. Xenografts showed a more severe rejection than allografts, with a marked inflammatory cell reaction throughout the graft. In contrast, in mice treated with the 5 mg/kg/day dose, xenografts exhibited a mild cell reaction and a greater number of regenerated myelinated fibers. In conclusion, effective axonal regeneration is achieved with FK506 administration at doses of 5 mg/kg/day through allografts and, partially, through xenografts.

Ulkur, E., F. Yuksel, et al. (2003). "Comparison of functional results of nerve graft, vein graft, and vein filled with muscle graft in end-to-side neurorrhaphy." Microsurgery 23(1): 40-8.
 End-to-side neurorrhaphy is an alternative method in the situation where the proximal part of the nerve cannot be found. When the intact nerve is not close enough to perform end-to-side neurorrhaphy, it will be necessary to use a graft for transporting the regenerating axons. In this study, we tried to find out whether it is possible to use a graft in an end-to-side neurorrhaphy, and compared the nerve graft with possible alternative grafts, i.e., vein and muscle-filled vein grafts. Thirty male Sprague Dawley rats were used, with an average weight of 293 g (range, 250-350 g). All experiments were done on the right side. A 2-cm nerve graft, beginning 1 cm distal to the branching level, was sectioned from the peroneal nerve. A 1-mm epineural window was opened in the tibial nerve. In the first group, the proximal side of this graft was sutured to the tibial nerve side in an end-to-side fashion, and the distal side was sutured to the distal peroneal nerve stump in an end-to-end fashion. In the second group, the right 2-cm jugular vein was harvested, and was used to bridge the defect instead of the nerve graft used in the first group. In the third group, a 2-cm jugular vein filled with fresh skeletal muscle was used to bridge the defect. At 2, 4, 8, 12, 20, and 28 weeks, functional assessment of nerve regeneration was performed, using walking-track analysis. The numbers of myelinated fibers and fiber diameters were measured, and an electron microscopic evaluation was carried out. Based on walking-track analysis and fiber diameters, the differences of all three groups were statistically significant (P < 0.05). While the differences of myelinated fibers between the first and second groups were not significant, the differences between the rest (group 1-group 3 and group 2-group 3) were significant (P < 0.05). Our study showed that, in end-to-side neurorrhaphy, the use of a nerve graft is possible, and a vein graft is also a good alternative, but a muscle-filled vein graft is not.

Ulmer, J. L., H. G. Krouwer, et al. (2003). "Pseudo-reorganization of language cortical function at fMR imaging: a consequence of tumor-induced neurovascular uncoupling." AJNR Am J Neuroradiol 24(2): 213-7.
 A left-handed patient with a grade II left frontal lobe astrocytoma had spontaneous seizures causing speech arrest and uncontrolled right upper extremity movements. Word-generation functional MR (fMR) imaging showed activity nearly exclusively in the right inferior frontal gyrus. The clinical history of the speech arrest and the intraoperative mapping proved left-hemisphere language dominance. Tumor involvement of the left inferior frontal gyrus caused uncoupling of the blood oxygenation level-dependent (BOLD) and neuronal response, leading to the erroneous fMR imaging appearance of right-hemisphere language dominance. Discrepancies between BOLD and intraoperative mapping in areas near lesions illustrate the complementary nature of these techniques.

Ulrich-Lai, Y. M., A. I. Fraticelli, et al. (2003). "Capsaicin-sensitive nerve fibers: a potential extra-ACTH mechanism participating in adrenal regeneration in rats." Microsc Res Tech 61(3): 252-8.
 Pituitary-derived factors, including ACTH, have been widely implicated in initiating adrenal regeneration. However, recent work has demonstrated that adrenal regeneration is also modulated by adrenal nerves that extensively reinnervate the regenerating adrenal. Moreover, transection of the splanchnic nerve removes sensory calcitonin gene-related peptide (CGRP) and preganglionic sympathetic vesicular acetylcholine transporter (VAChT)-positive fibers from the regenerating gland and delays regeneration. However, it is not known whether the splanchnic nerve effects on adrenal regeneration are mediated by the CGRP-positive or VAChT-positive innervation. The present studies use the drug capsaicin, a neurotoxin selective for a subset of primary afferent neurons, to specifically remove CGRP-positive fibers from the adrenal gland and assess subsequent effects on the recovery of adrenal mass and function after surgical enucleation. Male, Sprague-Dawley rats were anesthetized and treated with capsaicin (vs. vehicle) periaxonally to the thoracic splanchnic nerve (33 mM, 15 minutes) or systemically (30-100 mg/kg for 4 days, s.c.). After 7-12 days of recovery, rats received right adrenalectomy and left adrenal enucleation. At 14 and 21 days postenucleation, prestress and poststress plasma and adrenals glands were collected; adrenals were weighed and fixed for immunolabeling of CGRP-positive nerve fibers. Periaxonal capsaicin treatment decreased adrenal CGRP content prior to surgical enucleation; however, reinnervation by CGRP-positive fibers was not prevented and regeneration was not affected. Systemic capsaicin treatment attenuated the reinnervation by CGRP-positive fibers and increased the rate, but not extent, of adrenal regeneration. These results support the hypothesis that adrenal innervation represents an extra-ACTH mechanism to modulate the rate of adrenal regeneration.

Uno, T., K. Shogaki, et al. (2003). "Growth associated protein-43 mRNA expression in nucleus ambiguus motoneurons after recurrent laryngeal nerve injury in the rat." Acta Otolaryngol 123(2): 292-6.
 OBJECTIVE: To investigate the time-dependent differences in growth associated protein-43 (GAP-43) mRNA expression in the nucleus ambiguus (NA) motoneurons after recurrent laryngeal nerve (RLN) transection in the rat. MATERIAL AND METHODS: Animals were sacrificed on Days 1, 7, 14 and 21 after axotomy. The sections were then processed for in situ hybridization of GAP-43 mRNA. RESULTS: GAP-43 mRNA transiently increased after axotomy, reaching a peak on postoperative Day 7, and then decreased gradually at 14 and 21 days after axotomy. The peak period of GAP-43 expression in the NA is different from those in the facial and hypoglossal nuclei after axotomy. CONCLUSION: We have already reported that neuronal nitric oxide synthase (nNOS) is induced in the NA motoneurons after axotomy and reaches a peak on postoperative Day 14. This study shows that the expressions of GAP-43 and nNOS are chronologically different.

Utvag, S. E., O. Grundnes, et al. (2003). "Influence of extensive muscle injury on fracture healing in rat tibia." J Orthop Trauma 17(6): 430-5.
 OBJECTIVES: The treatment of tibial fractures associated with severe soft tissue injury remains a challenge. The objective of our experiment was to ascertain the influence of standardized muscle injuries on fracture healing in a nailed rat tibial fracture model. We hypothesized that a severe crush injury of leg muscles might not be as deleterious to fracture healing as total loss of a large muscle segment. STUDY DESIGN: A randomized study in male Wistar rats with a diaphyseal osteotomy. METHODS: Three separate, but complementary experiments were done in 51 rats. The first experiment involved 30 rats randomly assigned to three increasingly severe soft tissue interventions in a nailed tibial osteotomy model. The second experiment involved 14 rats divided into two groups to study blood flow measurements of the muscle envelope after soft tissue injury. Seven rats were used in the third experiment to provide biomechanical data and dimensions of the rat tibia. The protocol for the first experiment was intramedullary nailing after a middiaphyseal osteotomy of the left tibia. In group A, the soft tissue injury was minimal, while the muscles in the anterolateral compartment were crushed in group B. Resection of the anterolateral compartment muscles, resulting in only skin coverage at the fracture site, was performed in group C. The fibular nerve was resected in all three groups so that the animals were non-weight bearing on the operated extremity. At 4 weeks, the healing bones in each group were studied clinically, radiographically, and biomechanically. RESULTS: Radiographs in two planes revealed a clearly visible fracture line in the three intervention groups at 4 weeks. The callus area following muscle resection in group C was significantly reduced compared with the minimal soft tissue injury in group A. Biomechanically, resection of the anterolateral compartment muscles in group C reduced maximum bending load and fracture energy compared with fractures with minimal soft tissue injury in group A, while bending rigidity and fracture energy was reduced compared with muscle crush in group B. No difference in mechanical characteristics was detected between the healing bones in groups A and B. CONCLUSION: This animal study indicates that crushing of the leg muscle envelope with reduced blood flow does not influence the quality of bone healing at 4 weeks to a significant degree. Resection of a large muscle segment impairs tibial fracture healing significantly.

Valverde, F. (2003). "[The olfactory bulb as a model for experimental studies]." Neurologia 18(4): 177-86.
 The olfactory bulb represents a model for several research works carried out in our laboratory. Several details concerning structure, cell varieties and connections have been described. Special attention should be given to the ensheathing glia which accompanies olfactory axons, it being responsible for its plasticity and regeneration, as well as intervening in glomerular formation and their specificity. The olfactory system also represents a good model for the study of certain developmental aspects of the nervous system, such as the formation of nerve tracts. The lateral olfactory tract extends into the basal telencephalon and its development is supported by the existence of pre-existing routes and several attractive or repulsive factors. Its development requires complex molecular interactions in combination with several signalling influences. Pax6 is a transcription factor required for brain development. Nasal structures and eyes are absent in the homozygous mutant mice. In this mutant we have described the presence of one particular structure which could be a prospective olfactory bulb. Specific cell markers have been used to demonstrate that this structure shows many structural characteristics of the olfactory bulb. These observations favor the hypothesis of the existence of specific patterns delimiting specific functional domains in absence of their proper afferent fibers.

van Adel, B. A., C. Kostic, et al. (2003). "Delivery of ciliary neurotrophic factor via lentiviral-mediated transfer protects axotomized retinal ganglion cells for an extended period of time." Hum Gene Ther 14(2): 103-15.
 Ciliary neurotrophic factor (CNTF) has recently been demonstrated to be one of the most promising neurotrophic factors to improve both the survival and regeneration of injured retinal ganglion cells (RGCs). In the present study, we used optic nerve transection as an in vivo model to evaluate the effectiveness of a self-inactivating, replication-deficient lentiviral-mediated transfer of human ciliary neurotrophic factor (SIN-PGK-CNTF) on the survival of axotomized adult rat RGCs. Counts of dextran-fluorescein isothiocyanate conjugated (D-FITC)-retrogradely labeled RGCs revealed that the percentage of RGCs was drastically reduced (<90% cell death) 21 days after optic nerve transection. Retinal sections stained with X-gal revealed that intravitreal injection of the control LacZ-expressing lentiviral vector (LV-LacZ) resulted in the transduction of RGCs and retinal pigment epithelium (RPE) cells. A single intravitreal injection of LV-CNTF at the time of axotomy significantly enhanced RGC survival at 14 and 21 days postaxotomy compared to controls. These results demonstrate for the first time that rapid and prolonged delivery of CNTF using lentiviral-mediated gene transfer to the retina is an effective treatment for rescuing axotomized RGCs for an extended period of time. These results suggest that early and continuous administration of CNTF could serve as a potential treatment for retinal disorders involving optic neuropathy and RGC injury such as in glaucoma.

Van der Zee, C. E., T. Y. Man, et al. (2003). "Delayed peripheral nerve regeneration and central nervous system collateral sprouting in leucocyte common antigen-related protein tyrosine phosphatase-deficient mice." Eur J Neurosci 17(5): 991-1005.
 Cell adhesion molecule-like receptor-type protein tyrosine phosphatases have been shown to be important for neurite outgrowth and neural development in several animal models. We have previously reported that in leucocyte common antigen-related (LAR) phosphatase deficient (LAR-deltaP) mice the number and size of basal forebrain cholinergic neurons, and their innervation of the hippocampal area, is reduced. In this study we compared the sprouting response of LAR-deficient and wildtype neurons in a peripheral and a central nervous system lesion model. Following sciatic nerve crush lesion, LAR-deltaP mice showed a delayed recovery of sensory, but not of motor, nerve function. In line with this, neurofilament-200 immunostaining revealed a significant reduction in the number of newly outgrowing nerve sprouts in LAR-deltaP animals. Morphometric analysis indicated decreased axonal areas in regenerating LAR-deltaP nerves when compared to wildtypes. Nonlesioned nerves in wildtype and LAR-deltaP mice did not differ regarding myelin and axon areas. Entorhinal cortex lesion resulted in collateral sprouting of septohippocampal cholinergic fibres into the dentate gyrus outer molecular layer in both genotype groups. However, LAR-deltaP mice demonstrated less increase in acetylcholinesterase density and fibre number at several time points following the lesion, indicating a delayed collateral sprouting response. Interestingly, a lesion-induced reduction in number of (septo-entorhinal) basal forebrain choline acetyltransferase-positive neurons occurred in both groups, whereas in LAR-deltaP mice the average cell body size was reduced as well. Thus, regenerative and collateral sprouting is significantly delayed in LAR-deficient mice, reflecting an important facilitative role for LAR in peripheral and central nervous system axonal outgrowth.

Van Helden, H. P., H. C. Trap, et al. (2003). "Long-term, low-level exposure of guinea pigs and marmosets to sarin vapor in air: lowest observable effect level." Toxicol Appl Pharmacol 189(3): 170-9.
 Realistic scenarios for low-level exposure to nerve agents will often involve exposures over several hours to extremely low doses of agent. In order to expose animals to the lowest controllable concentrations of agent and to increase exposure times until a lowest observable effect level (LOEL) becomes measurable, a validated system was developed for exposing conscious animals to 0.05-1.0 microg/m(3) (8-160 ppt) of sarin and other nerve agents. Based on cold trapping of sarin from the exposure air, the concentration could be measured semicontinuously, at 4-min time intervals by means of gas chromatography. We found that the LOEL upon a 5-h whole body exposure of guinea pigs and marmosets to sarin vapor corresponds with the measurement of an internal dose by means of fluoride-induced regeneration of sarin from phosphylated binding sites in plasma, mostly BuChE. For guinea pigs the LOEL was observed at Ct = 0.010 +/- 0.002 mg/min/m(3), whereas a Ct of 0.04 +/- 0.01 mg/min/m(3) was established for the LOEL in marmosets. These levels are several orders of magnitude lower than those based on classical measurement of depressed cholinesterase activities. At low exposure levels of guinea pigs and marmosets (< or =1 microg/m(3)), a reasonable linearity was observed between exposure dose and internal dose. The data were addressed in the light of the recently recommended occupational exposure limits to sarin for workers without respiratory protection, which suggests that the exposure limits should be reconsidered if the slightest inhibition of cholinesterases should be prevented.

van Ooij, A., R. Weijers, et al. (2003). "Remodelling of the sacrum in high-grade spondylolisthesis: a report of two cases." Eur Spine J 12(3): 332-8.
 Two young patients are described, who were operated on for high-grade spondylolisthesis. A good posterolateral fusion was achieved, without decompression and without reduction. The clinical course was favourable, the tight hamstring syndrome resolved. Disappearance of the posterior-superior part of the sacrum and of the posterior part of the L5-S1 disc was observed on comparing pre- and postoperative magnetic resonance (MR) images. This resulted in normalisation of the width of the spinal canal. Around the L5 nerve roots in the L5-S1 foramina some fat reappeared. These anatomical changes on MRI could play a role in the disappearance of clinical symptoms.

Vanmali, B. H., E. V. Romanova, et al. (2003). "Endogenous neurotrophic factors enhance neurite growth by bag cell neurons of Aplysia." J Neurobiol 56(1): 78-93.
 Mechanisms that regulate neurite outgrowth are phylogenetically conserved, including the signaling molecules involved. Here, we describe neurotrophic effects on isolated bag cell neurons (BCNs) of substrate-bound growth factors endogenous to the sea slug Aplysia californica. Sheath cells dissociated from the pleural-visceral connectives of the Aplysia CNS and arterial cells dissociated from the anterior aorta enhance neurite outgrowth when compared to controls, i.e., BCNs grown in defined medium alone. In addition, the substrate remaining after sheath cells or arterial cells are killed significantly enhances growth, relative to all other conditions tested. For instance, primary neurites are more numerous and greater in length for BCNs cultured on substrate produced by arterial cells. These results suggest that sheath and arterial cells produce growth-promoting factors, some of which are found in the substrates produced by these cell types. Using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), we found that Aplysia collagen-like peptides are produced by dissociated arterial cells, and therefore likely contribute to the observed growth effects. Collagen-like peptides and other factors produced by sheath and arterial cells likely influence neurite growth in the Aplysia CNS during development, learning and memory, and regeneration after injury.

Varejao, A. S., A. M. Cabrita, et al. (2003). "Nerve regeneration inside fresh skeletal muscle-enriched synthetic tubes: a laser confocal microscope study in the rat sciatic nerve model." Ital J Anat Embryol 108(2): 77-82.
 The search for good conduits for bridging nerve defects is a major challenge of today's tissue engineering research. In this paper we report on a laser confocal microscope study on early nerve regeneration inside a tissue engineered graft made by a poly(DLLA-epsilon-CL) conduit enriched with fresh skeletal muscle. The same biodegradable tubes filled with PBS solution were used as controls. The conduits were placed to bridge unilateral 1-cm-long rat sciatic nerve defects and analysed 10 days after surgery. Results showed that inside the muscle-enriched tubes axon regeneration, labelled by means of anti-neurofilament antibody, was already begun, whilst no axon regeneration was detectable along control tubes. In addition, a-GFAP (glial fibrillar acid protein) immuno-labelling of Schwann cells showed that progression inside muscle-enriched tubes, especially from the distal nerve stump, was much more evident than in control conduits. These results suggest that enrichment of synthetic tubes with fresh skeletal muscle promotes axon regeneration and Schwann cell migration in early nerve repair stages.

Varejao, A. S., A. M. Cabrita, et al. (2003). "Morphology of nerve fiber regeneration along a biodegradable poly (DLLA-epsilon-CL) nerve guide filled with fresh skeletal muscle." Microsurgery 23(4): 338-45.
 Previous morphological and morphometrical studies showed that fresh-skeletal-muscle-enriched vein segments are good conduits for leading peripheral nerve regeneration. In the present study, we investigated the morphological features of peripheral nerve fibers regenerated along a 10-mm-long biodegradable poly (DLLA-epsilon-CL) nerve guide enriched with fresh skeletal muscle, comparing them to nerve fiber regeneration along 10-mm-long phosphate-buffered saline (PBS)-enriched poly (DLLA-epsilon-CL) tubes. Repaired nerves were analyzed at weeks 6 and 24 postoperatively. Structural and ultrastructural observation showed that good nerve fiber regeneration occurred in both PBS-enriched and fresh-skeletal-muscle-enriched nerve guides, and histomorphometrical analysis of regenerated myelinated fibers revealed no statistically significant differences between the two experimental groups at week 24 after surgery. The employment of fresh-muscle-enriched conduits for the repair of nerve defects is critically discussed in the light of these results.

Verdu, E., G. Garcia-Alias, et al. (2003). "Olfactory ensheathing cells transplanted in lesioned spinal cord prevent loss of spinal cord parenchyma and promote functional recovery." Glia 42(3): 275-86.
 We studied the effects of olfactory ensheathing cells (OECs) transplanted in a photochemical spinal cord injury in adult rats. After dorsal laminectomy at T8 vertebra, subjacent spinal cord was bathed with rose Bengal for 10 min and illuminated with visible light by means of an optic fiber connected to a halogen lamp for 2.5 min at maximal intensity of 8 kLux. Eight injured rats received a suspension of OECs in DMEM, and another eight rats received DMEM alone. Locomotor ability scored by the BBB scale, pain sensibility by the plantar algesimetry test, and motor- and somatosensory-evoked potentials by electrophysiological techniques were evaluated for 3 months postsurgery. Finally, all rats were perfused with paraformaldehyde and transverse sections from the spinal cord segment at the lesion site were immunostained against GFAP. Area of the preserved spinal cord parenchyma was measured from the GFAP-immunolabeled cord sections. The BBB score and the amplitude of motor- and somatosensory-evoked potentials were higher in OECs-transplanted rats than in DMEM-injected animals throughout follow-up, whereas the withdrawal response to heat noxious stimulus was lower in OEC- than in DMEM-injected rats. The area of preserved spinal cord was significantly larger in OECs-transplanted rats than in DMEM-injected animals. These results indicate that OECs promote functional and morphological preservation of the spinal cord after photochemical injury.

Vergnolle, N., M. Ferazzini, et al. (2003). "Proteinase-activated receptors: novel signals for peripheral nerves." Trends Neurosci 26(9): 496-500.
 The discovery of proteinase-activated receptors (PARs) in the nervous system has led to new insights about the potential physiological functions of these enzymes, which were traditionally considered merely as degradative molecules. This review summarizes evidence that proteinases, through activation of PARs, interact with the peripheral nervous system (PNS), playing roles in neurogenic inflammation, pain perception, secretory and motor functions, as well as in the response to nerve injuries. Activation of PARs interferes with numerous physiological events that are under tight neural control, in addition to modulating nerve survival. New potential roles are suggested for members of the PAR family, highlighting proteinases and their receptors as potential therapeutic targets for diseases associated with PNS activation.

Vernadakis, A. J., H. Koch, et al. (2003). "Management of neuromas." Clin Plast Surg 30(2): 247-68, vii.
 After nerve injury and regeneration, significant pain may be associated with the scar and altered sensation observed within the distribution of the injured nerve. A bulbous swelling may form at the severed nerve end, constituting a traumatic neuroma. The development of a painful neuroma may be more disabling to the patient than an area of anesthesia or even loss of motor function. Effective treatment of the painful neuroma remains a difficult problem. Diminished productivity, alterations in patient lifestyle, and possible progression to chronic pain syndromes must be considered within the scope of neuroma management, and treatment must focus on alleviating the pain and restoring the functional loss caused by the nerve injury. Careful patient selection is the cornerstone of successful outcomes. Once the patient has been selected, the surgical management of the painful neuroma throughout the body is based on basic principles that vary only slightly from region to region. Using these tenets, a neuroma management algorithm has been developed based on the pathophysio-logy of the neuroma, the results of experimental studies, review of patient outcomes, and understanding the psychology of pain in the surgical patient.

Verze, L., A. Paraninfo, et al. (2003). "Expression of neuropeptides and growth-associated protein 43 (GAP-43) in cutaneous and mucosal nerve structures of the adult rat lower lip after mental nerve section." Ann Anat 185(1): 35-44.
 The reinnervation of the adult rat lower lip has been investigated after unilateral section of the mental nerve. Rats were sacrificed at 4, 7, 9, 14, 30, and 90 days after the operation. A further group of animals with section of the mental nerve and block of the alveolar nerve regeneration, was sacrificed at 14 days. Specimens were processed for immunocytochemistry with antibodies against PGP 9.5, GAP-43 or neuropeptides (CGRP, SP and VIP). Four days after nerve section, axonal degeneration seems evident in the mental nerve branches and inside skin and mucosa. GAP-43 immunoreactivity is intense in the mental nerve 7 days after nerve section and it reaches its maximal expression and distribution in peripheral nerve fibres at 14 days. At 30 days, the decline in its expression is associated with the increase of PGP9.5-, SP-, and CGRP immunopositivity. VIP is observed only in perivascular fibres at all times observed. Present results suggest that, after sensory denervation of the rat lip, nerve fibres in skin and mucosa remain at lower density than normal. The different time courses in the expression of neuropeptides and GAP-43 suggest a possible early involvement of GAP-43 in peripheral nerve regeneration.

Vinke, J. G., J. J. Hoorweg, et al. (2003). "[Trigeminal trophic syndrome: 2 patients with a non-healing ulcer on the ala nasi]." Ned Tijdschr Geneeskd 147(38): 1866-9.
 Two female patients, aged 74 and 91 years respectively, had a skin ulcer on a nostril and ipsilateral sensory paralysis in the area covered by the N. trigeminus. This is known as trigeminal trophic syndrome. It may be caused by (iatrogenic) damage to the trigeminal nerve by chronic manipulation of the senseless skin. Only in case of doubt, histological examination of a tissue sample is advised to exclude malignancy. The main treatment consists of instructing the patient not to manipulate the skin. In some patients surgical reconstruction is necessary.

Vogelaar, C. F., M. F. Hoekman, et al. (2003). "Homeobox gene expression in adult dorsal root ganglia during sciatic nerve regeneration: is regeneration a recapitulation of development?" Eur J Pharmacol 480(1-3): 233-50.
 After damage of the sciatic nerve, a regeneration process is initiated. Neurons in the dorsal root ganglion regrow their axons and functional connections. The molecular mechanisms of this neuronal regenerative process have remained elusive, but a relationship with developmental processes has been conceived. This chapter discusses the applicability of the developmental hypothesis of regeneration to the dorsal root ganglion; this hypothesis states that regeneration of dorsal root ganglion neurons is a recapitulation of development. We present data on changes in gene expression upon sciatic nerve damage, and the expression and function of homeobox genes. This class of transcription factors plays a role in neuronal development. Based on these data, it is concluded that the hypothesis does not hold for dorsal root ganglion neurons, and that regeneration-specific mechanisms exist. Cytokines and the associated Jak/STAT (janus kinase/signal transducer and activator of transcription) signal transduction pathway emerge as constituents of a regeneration-specific mechanism. This mechanism may be the basis of pharmacological strategies to stimulate regeneration.

Volozhin, A. I., N. D. Brusenina, et al. (2003). "[Mechanisms disordering wound healing on the lip after bilateral crossing of the inferior alveolar nerve and experimental validation of correction methods]." Stomatologiia (Mosk) 82(2): 4-9.
 The mechanisms of lip wound healing after bilateral crossing of the inferior alveolar nerve (IAN) were studied on Chinchilla rabbits in 3 experimental series, 6 animals per series. In group 1 bilateral crossing of IAN was carried out, in group 2 bilateral crossing of IAN was paralleled by removal of a mucous flap in the middle of the lower lip, and in group 3 the same wound as in group 2 was created, after which the wounds in this group were daily treated with a special ointment and a single injection of lidocaine (1% solution) under the wound. The nerve crossing led to development of ulcer on the lip with degenerative changes in the vascular walls, destruction of nerve fibers, and fragmentation of some axial cylinders. Crossing of IAN simultaneously with removal of the lower lip flap led to more severe degenerative changes in the tissue. Daily treatment of the lip with the ointment and lidocaine blocking normalized wound healing. A possible mechanism of the changes observed is discussed.

Vroemen, M., L. Aigner, et al. (2003). "Adult neural progenitor cell grafts survive after acute spinal cord injury and integrate along axonal pathways." Eur J Neurosci 18(4): 743-51.
 The main rationale for cell-based therapies following spinal cord injury are: (i) replacement of degenerated spinal cord parenchyma by an axon growth supporting scaffold; (ii) remyelination of regenerating axons; and (iii), local delivery of growth promoting molecules. A potential source to meet these requirements is adult neural progenitor cells, which were examined in the present study. Fibroblast growth factor 2-responsive adult spinal cord-derived syngenic neural progenitor cells were either genetically modified in vitro to express green fluorescent protein (GFP) using retroviral vectors or prelabelled with bromodeoxyuridine (BrdU). Neural progenitor cells revealed antigenic properties of neurons and glial cells in vitro confirming their multipotency. This differentiation pattern was unaffected by retroviral transduction. GFP-expressing or BrdU-prelabelled neural progenitor cells were grafted as neurospheres directly into the acutely injured rat cervical spinal cord. Animals with lesions only served as controls. Three weeks postoperatively, grafted neural progenitor cells integrated along axonal profiles surrounding the lesion site. In contrast to observations in culture, grafted neural progenitor cells differentiated only into astro- and oligodendroglial lineages, supporting the notion that the adult spinal cord provides molecular cues for glial, but not for neuronal, differentiation. This study demonstrates that adult neural progenitor cells will survive after transplantation into the acutely injured spinal cord. The observed oligodendroglial and astroglial differentiation and integration along axonal pathways represent important prerequisites for potential remyelination and support of axonal regrowth.

Vroemen, M. and N. Weidner (2003). "Purification of Schwann cells by selection of p75 low affinity nerve growth factor receptor expressing cells from adult peripheral nerve." J Neurosci Methods 124(2): 135-43.
 The intrinsic capacity of Schwann cells to promote regeneration after limited peripheral nerve lesions has been successfully transferred to extensive peripheral nerve injuries and central nervous system lesions by autologous transplantation strategies. However, both the intrinsic ability of axotomized neurons to regenerate and the permissiveness of the parenchyma surrounding the acute injury site diminish over time. Therefore, the autologous transplantation mode requires a fast and effective method to isolate Schwann cells from peripheral nerve biopsies. Here, we report a method to purify p75 low affinity nerve growth factor receptor (p75LNGFr) expressing Schwann cells from peripheral nerve biopsies in adult rats using magnetic-activated cell separation (MACS). After 1 week of nerve degeneration in culture, nerve fragments were dissociated resulting in mixed cultures containing Schwann cells and fibroblasts. After incubation with specific anti-p75LNGFr antibodies and secondary magnetic bead conjugated antibodies followed by one cycle of MACS, 95% pure Schwann cell cultures were generated as confirmed by flow-cytometry and immunocytochemistry. In contrast to established methods, MACS separation of p75LNGFr expressing cells allows the reliable purification of Schwann cells within 9 days after biopsy employing direct selection of Schwann cells rather than fibroblast depletion assays. Therefore, this method represents an effective and fast means to generate autologous Schwann cells for clinical transplantation strategies aiming for axon repair and remyelination.

Waddell, R. L., K. G. Marra, et al. (2003). "Using PC12 Cells To Evaluate Poly(caprolactone) and Collagenous Microcarriers for Applications in Nerve Guide Fabrication." Biotechnol Prog 19(6): 1767-1774.
 Tissue-engineered nerve guides can provide mechanical support as well as chemical stimulation for nerve regeneration. PC12 cells were used to test the novel combination of poly(caprolactone) (PCL) and macroporous collagen-based microcarriers (CultiSphers) as an initial phase in the fabrication of multichanneled nerve guides. Laminin-coated PCL was an effective matrix for the attachment, proliferation, and viability of PC12 cells, relative to uncoated PCL. PC12 cells attached to laminin-coated PCL and extended neurites when cultured in the presence of nerve growth factor (NGF). PC12 cells attached and proliferated on CultiSphers and extended neurites in response to NGF. A novel PCL/CultiSpher composite material also supported PC12 attachment and proliferation and provides a potentially useful material for the fabrication of synthetic nerve guides.

Wakisaka, S. and Y. Atsumi (2003). "Regeneration of periodontal Ruffini endings in adults and neonates." Microsc Res Tech 60(5): 516-27.
 We reviewed the regeneration of periodontal Ruffini endings, primary mechanoreceptors in the periodontal ligament, following injury to the inferior alveolar nerve (IAN) in adult and neonatal rats. Morphologically, mature Ruffini endings are characterized by an extensive arborization of axonal terminals and association with specialized Schwann cells, called lamellar or terminal Schwann cells. Following injury to IAN in the adult, the periodontal Ruffini endings of the rat lower incisor ligament regenerate more rapidly than Ruffini endings in other tissues. During regeneration, terminal Schwann cells migrate into regions where they are never found under normal conditions. The development of periodontal Ruffini endings of the rat incisor is closely associated with the eruption of the teeth; the morphology and distribution of the terminal Schwann cells became almost identical to those in adults during postnatal days 15-18 (PN 15-18d) when the first molars appear in the oral cavity, while the axonal elements showed extensive ramification around PN 28d when the functional occlusion commences. When the IAN was injured in neonates, the regeneration of periodontal Ruffini endings was delayed compared with the adults. The migration of terminal Schwann cells is also observed following IAN injury, after which the distribution of terminal Schwann cells became almost identical to that of the adults, i.e., PN 14d. Since the interaction between axon and Schwann cell is important during regeneration and development, further studies are required to elucidate its molecular mechanism during the regeneration as well as the development of the periodontal Ruffini endings.

Walbeehm, E. T., E. B. Dudok van Heel, et al. (2003). "Nerve compound action current (NCAC) measurements and morphometric analysis in the proximal segment after nerve transection and repair in a rabbit model." J Peripher Nerv Syst 8(2): 108-15.
 In the evaluation of nerve regeneration using magneto-neurography (MNG), the proximal segment showed a reproducible decrease in peak-peak amplitude of the nerve compound action current's (NCAC) of 60%. To explain these changes, morphometry of myelinated axons in the proximal segment is compared to the MNG signals. A standardised nerve transection and reconstruction was performed in rabbits. NCACs were measured approximately 5 cm proximal to the lesion from operated and control nerves after 12 weeks. Histological samples were taken from the same area of the nerve where the NCACs were obtained. Results showed a decrease of the peak-peak amplitude of the NCAC of 57% compared to the control. Conduction velocity decreased 15% (not significant). Morphometry elicited a decrease in larger (10-15 microm) axons (284 +/- 134 vs 82 +/- 55) and an increase in smaller (2-5 microm) axons (1445 +/- 360 vs 1921 +/- 393). A strong correlation existed between the decrease in amplitude and the decrease in larger axons (0.85). Peak-peak amplitude varies approximately with the square of the diameter axon. Therefore, because peak-peak amplitude is mainly dependent on the larger-diameter axons, the decrease in peak-peak amplitude of the NCACs may be explained by a decrease in numbers of 10-15-microm axons.

Wang, J., N. M. Goodger, et al. (2003). "A method of invaginating the facial vein for inferior alveolar nerve repair." J Oral Maxillofac Surg 61(7): 848-9.
 
Wang, S., Q. Cai, et al. (2003). "Acceleration effect of basic fibroblast growth factor on the regeneration of peripheral nerve through a 15-mm gap." J Biomed Mater Res 66A(3): 522-31.
 In this study, nerve guides composed of poly(D,L-lactide) (PDLLA) were fabricated and used in the repair of transected sciatic nerves (15-mm gaps) of rats. Nerve guides with a two-ply structure (inner layer dense, outer layer microporous) were prepared by controlling the solvent evaporation rate. Then basic fibroblast growth factor (bFGF) was embedded in the inner layer of the nerve guides. Thus the inner dense layer not only could prevent the ingrowth of fibroblast and avoid the outgrowing nerve cable, but it also could retain the released bFGF in the guide lumen. The outer porous layer allowed vascular ingrowth and the diffusion of essential nutrients into the guide lumen. The data show that by using this nerve guide, the transected 15-mm sciatic nerve was regenerated successfully within 4 months. The recovery of function of the regenerated nerves was significantly accelerated by bFGF, as indicated by an electrostimulation test and histologic assays. In addition, the bFGF retained its bioactivity during embedding and continuously was released from the matrix, as confirmed by the results of both the dorsal root ganglia (DRG) and the Schwann cell culture in the presence of PDLLA matrix containing bFGF. The released bFGF enhanced the ability of the nerve fibers to sprout from dorsal root ganglia, and it accelerated the proliferation of Schwann cells.

Wang, Y. Z., J. H. Meng, et al. (2003). "Differentiation-inducing and protective effects of adult rat olfactory ensheathing cell conditioned medium on PC12 cells." Neurosci Lett 346(1-2): 9-12.
 Transplantation of olfactory ensheathing cells has been one of the promising strategies in enhancing central nerve fiber regeneration. Membrane surface molecules on olfactory ensheathing cells mediating cell-cell interactions as well as various factors released from them are thought to be important for nerve regeneration. The latter, however, has not been fully substantiated by experimental data, particularly regarding the olfactory ensheathing cells of the adult animals. In the present study, the effects of adult olfactory ensheathing cell conditioned medium on PC12 cells were examined. The results show that the factors secreted by the adult olfactory ensheathing cells can promote PC12 cell differentiation and protect it from Zn(2+) insult.

Wankell, M., S. Werner, et al. (2003). "The roles of activin in cytoprotection and tissue repair." Ann N Y Acad Sci 995: 48-58.
 Activin is a member of the transforming growth factor beta family of growth and differentiation factors. Initially discovered as a protein that stimulates release of follicle-stimulating hormone, it is now well accepted as an important regulator of cell growth and differentiation. Most interestingly, a series of previous studies have revealed novel roles of activin in inflammation and repair. Our own results have provided evidence for an important function of activin in cutaneous wound repair as well as in neuroprotection, and these data will be summarized and discussed in this chapter.

Wang, J. F., M. E. Olson, et al. (2003). "Recombinant connective tissue growth factor modulates porcine skin fibroblast gene expression." Wound Repair Regen 11(3): 220-9.
 Connective tissue growth factor (CTGF) is a 38 Kda cysteine-rich, heparin-binding peptide that has been implicated in several normal and abnormal physiological processes. CTGF has been shown to be induced by transforming growth factor-beta. Previous studies in our pig model of skin wound healing showed a coordinate expression of transforming growth factor-beta and CTGF during the healing process. To better understand the function of CTGF during wound healing, normal porcine fibroblasts were isolated from skin samples from SPF Yorkshire pigs. At fourth passage the cells were cultured in Dulbecco's modified Eagle's medium supplemented with fetal calf serum and at 80% confluence the medium was replaced with supplemented serum-free medium. After a further 24 hours, cells were treated with 0, 10, 25, 50, 100, and 500 ng/ml of 38 Kda or 16-20 Kda (C-terminal truncated form) recombinant expressed human CTGF for 24 hours or treated with 100 ng/ml for 0, 12, 24, and 48 hours. Subsequently, CTGF effects on cell DNA synthesis and mRNA levels for a subset of relevant molecules were assessed. The results showed that in cells treated with 38 Kda rhCTGF, mRNA levels for types I and III collagen, fibromodulin, and basic fibroblast growth factor were significantly up-regulated, but mRNA levels for HSP47, decorin, biglycan, and versican were not significantly altered. mRNA levels for CTGF were also significantly increased, indicating autoregulation of expression. However, mRNA levels for transforming growth factor-beta, inteleukins 1 and 6, tumor necrosis factor-alpha, and nerve growth factor did not change. Interestingly, mRNA levels for the tissue inhibitors of metalloproteinase-1, -2, -3 and -4 were observed to significantly increase, but in contrast, mRNA levels for matrix metalloproteinases-1, -2, -9 were not significantly altered by exposure of the cells to the 38 Kda form of CTGF. In addition, DNA synthesis was augmented in the presence of 38 Kda rhCTGF. However, the truncated 16-20 Kda form of rhCTGF appeared to have none of these effects on porcine fibroblasts. These results indicate that in order to induce changes in porcine fibroblasts a molecule with an intact C-terminal domain is required, and that CTGF regulates porcine fibroblast extracellular matrix molecule, growth factor, and proteinase inhibitor gene expression without apparently affecting matrix metalloproteinase mRNA levels. These findings suggest that CTGF contributes to the anabolic environment during skin wound healing via selective modulation of fibroblast proliferation and changes to gene expression.

Watanabe, M., Y. Tokita, et al. (2003). "Intravitreal injections of neurotrophic factors and forskolin enhance survival and axonal regeneration of axotomized beta ganglion cells in cat retina." Neuroscience 116(3): 733-42.
 Some retinal ganglion cells in adult cats survive axotomy for two months and regenerate their axons when a peripheral nerve is transplanted to the transected optic nerve. However, regenerated retinal ganglion cells were fewer than 4% of the total retinal ganglion cell population in the intact retina. The present study examined the effects of intravitreal injections of neurotrophic factors (brain-derived neurotrophic factor, ciliary neurotrophic factor, basic fibroblast growth factor, glial cell-derived neurotrophic factor, neurotrophin 4), first on the survival of axotomized cat retinal ganglion cells within 2 weeks, and then on axonal regeneration of the retinal ganglion cells for 2 months after peripheral nerve transplantation. We tested first enhancement of the survival by one of the factors, and then one or two of them supplemented with forskolin, which increases intracellular cAMP. Single injections of 0.5 microg or 1 microg brain-derived neurotrophic factor, 1 microg ciliary neurotrophic factor, or 1 microg glial cell-derived neurotrophic factor significantly increased total numbers of surviving retinal ganglion cells; 1.6-1.8 times those in control retinas. Identification of retinal ganglion cell types with Lucifer Yellow injections revealed that the increase of surviving beta cells was most conspicuous: 2.5-fold (brain-derived neurotrophic factor) to 3.6-fold (ciliary neurotrophic factor). A combined injection of 1 microg brain-derived neurotrophic factor, 1 microg ciliary neurotrophic factor, and 0.1 mg forskolin resulted in a 4.7-fold increase of surviving beta cells, i.e. 50% survival on day 14. On the axonal regeneration by peripheral nerve transplantation, a combined injection of brain-derived neurotrophic factor, ciliary neurotrophic factor, and forskolin resulted in a 3.4-fold increase of beta cells with regenerated axons. The increase of regenerated beta cells was mainly due to the enhancing effect of neurotrophic factors on their survival, and possibly to a change of retinal ganglion cell properties by cAMP to facilitate their axonal regeneration.

Webb, S., C. A. Munro, et al. (2003). "Is multicomponent T2 a good measure of myelin content in peripheral nerve?" Magn Reson Med 49(4): 638-45.
 Multicomponent T(2) relaxation of normal and injured rat sciatic nerve was measured. The T(2) relaxation was multiexponential, indicating the multicompartmental nature of T(2) decay in nerve tissue. The size of the short, observed T(2) component correlated very well with quantitative assessment of myelin using computer-assisted histopathological image analysis of myelin. Specifically, the size of the short T(2) component reflected the processes of myelin loss and remyelination accompanying Wallerian degeneration and regeneration following trauma. However, it represented all myelin present in the sample and did not distinguish between intact myelin and myelin debris. Other changes in T(2) spectra were also observed and could be correlated with axonal loss and inflammation. The study also questions the validity of previously offered interpretations of T(2) spectra of nerve.

Weerth, S. H., H. Rus, et al. (2003). "Complement C5 in experimental autoimmune encephalomyelitis (EAE) facilitates remyelination and prevents gliosis." Am J Pathol 163(3): 1069-80.
 Activation of the classical complement system is known to play a central role in autoimmune demyelination. We have analyzed the role of complement component C5 in experimental autoimmune encephalomyelitis (EAE) using C5-deficient (C5-d) and C5-sufficient (C5-s) mice. Both groups of mice displayed early onset EAE, a short recovery phase, and similar stable chronic courses. However, in contrast to the clinical similarities, marked differences were apparent by histopathology. During acute EAE in C5-d, a delay in inflammatory cell infiltration and tissue damage was observed along with restricted lesion areas, which in C5-s mice were more extensive and diffuse. More striking were the differences in chronic lesions. In C5-d mice, inflammatory demyelination and Wallerian degeneration were followed by axonal depletion and severe gliosis, while in C5-s, the same initial signs were followed by axonal sparing and extensive remyelination. In C5-d, immunohistochemistry and Western blotting showed an increase in glial fibrillary acidic protein and a decrease in neurofilament protein, proteolipid protein, and several pro-inflammatory markers. These results in the EAE model indicate that absence of C5 resulted in fiber loss and extensive scarring, whereas presence of C5-favored axonal survival and more efficient remyelination.

Wei, X., J. Lao, et al. (2003). "Bridging peripheral nerve defect with chitosan-collagen film." Chin J Traumatol 6(3): 131-4.
 OBJECTIVE: To seek new method for the treatment of peripheral nerve injury. METHODS: In rat model with sciatic nerve defect, chitosan-collagen film was sutured into conduit to bridge 5 mm, 10 mm nerve defects. Rats that underwent end-to-end anastomosis were taken as controls. General observation, electrophysiological study, histological study and image analysis were performed at 4, 8, 12 weeks postoperatively. RESULTS: In 5 mm nerve defects, the quality of nerve regeneration was similar to that of the control group. For 10 mm nerve defect, nerve regeneration was inferior to that of the control group. Chitosan-collagen film obviously degraded at 12 weeks postoperatively. CONCLUSIONS: Chitosan-collagen film conduit can be used to bridge peripheral nerve defect.

Westerlund, T., V. Vuorinen, et al. (2003). "The effect of combined neurolytic blocking agent 5% phenol-glycerol in rat sciatic nerve." Acta Neuropathol (Berl) 106(3): 261-70.
 Combined 5% phenol-glycerol has been used to treat cancer pain or spasticity and as sympathetic blocks. The major clinical problems have been the unpredictable effects on pain and on the duration of the blocks. Previously we have shown that intraneurally injected phenol induces haemorrhagic necrosis as well as dissolving of the nerve fibres. Glycerol, on the other hand, induces dispersion of nerve fibre debris into the endoneurium. We have now studied the effects of a combination of these two chemically different agents. The endoneurial and epineurial responses of rat peripheral nerve were followed after intraneural and perineural injections. Samples for electron microscopic and immunohistochemical studies were taken at 1-26 weeks after the injection. The intraneural phenol-glycerol injection resulted in gross endoneurial damage with partly or totally dissolved nerve fibres. Totally dissolved nerve fibres showed empty, collapsed basal lamina tubes and partly dissolved nerve fibres showed breaching of remaining degenerative debris into the endoneurial space. Axonal regeneration was delayed and was observed first after 2 weeks and it took 4 months before most of the nerve fibres were myelinated. The perineural injections resulted in partial subperineurial damage of the endoneurium morphologically similar to the results caused by the intraneural injection. An initial high accumulation of epineurial macrophages was noted at 1 and 2 weeks. An invasion of macrophages into the endoneurium occurred within 1 week after the intraneural and perineural injections and the number of endoneurial macrophages remained high for up to 6 months. The present study shows that glycerol added to phenol diminishes the necrotizing effect of phenol after an intraneural injection. Combined phenol-glycerol-induced nerve injury is reversible and the axons regenerate but residual morphological changes can be observed even after 6 months.

Wiberg, M. and G. Terenghi (2003). "Will it be possible to produce peripheral nerves?" Surg Technol Int 11: 303-10.
 Several hundred thousand peripheral nerve injuries occur each year in Europe and the United States alone, mainly as a result of trauma to the upper extremity. Even after optimal surgical repair, functional outcome-especially sensory recovery-is disappointingly poor. This poor outcome is largely due to death of primary sensory neurons, but also lack of fiber regeneration over the nerve trauma zone and target-organ denervation. The type of nerve repair performed is dependent on the size of the nerve gap between the proximal and distal stumps. Short gaps can be repaired directly by end-to-end coaptation and epineural suturing, whereas long nerve gaps require additional material to bridge the defect, which further reduces the functional outcome. The current repair method to bridge nerve defects is the use of autologous nerve grafts (autografts), which provide the regenerating axons with a natural guidance channel populated with functioning Schwann cells (SC) surrounded by their basal lamina. The first use of nerve grafts in humans was reported in 1878, but the wide use of this technique was developed during World War II when nerve grafting became the standard method for nerve-gap repair. Harvesting of nerve grafts results in co-morbidity that includes scarring, loss of sensation, and possible formation of painful neuroma. The donor nerves often are of small calibre and limited number. As functional recovery in peripheral nerve reconstruction is poor, clearly, alternative method for bridging nerve gaps is needed.

Wiberg, M., A. Hazari, et al. (2003). "Sensory recovery after hand reimplantation: a clinical, morphological, and neurophysiological study in humans." Scand J Plast Reconstr Surg Hand Surg 37(3): 163-73.
 Despite fairly good return of motor function, patients who have amputated hands reimplanted demonstrate poor sensory recovery and severe cold intolerance, two variables that are difficult to quantify reliably. In this study we wanted to find out if there is a correlation between morphological findings of sensory and sympathetic reinnervation and clinical and neurophysiological variables. Skin was biopsied from the reimplanted and corresponding area in the normal hands of eight patients who had sustained a hand amputation and subsequent reimplantation. The sections were immunostained using markers for both sensory and sympathetic nerve fibres. Comparison between the reimplanted and normal sides in each individual showed a mean loss of sensory immunoreactive nerve fibres of 30%, and for sympathetic immunoreactivity the loss was 60%. There was measurable two-point discrimination in the injured hand only in patients below the age of 40 years, corresponding to the better recovery of mechanical thresholds evaluated neurophysiologically for this age group. These results confirm the extensive loss of sensory nerve fibres after nerve injury, probably correlated to loss of sensory neurons. We have also shown that it is possible to correlate the results of clinical and neurophysiological evaluation with morphological results of skin reinnervation specific to the repaired nerve, and so improve the possibility for the quantification of sensory recovery.

Widenfalk, J., A. Lipson, et al. (2003). "Vascular endothelial growth factor improves functional outcome and decreases secondary degeneration in experimental spinal cord contusion injury." Neuroscience 120(4): 951-60.
 Spinal cord injury leads to acute local ischemia, which may contribute to secondary degeneration. Hypoxia stimulates angiogenesis through a cascade of events, involving angiogenesis stimulatory substances, such as vascular endothelial growth factor (VEGF). To test the importance of angiogenesis for functional outcome and wound healing in spinal cord injury VEGF165 (proangiogenic), Ringer's (control) or angiostatin (antiangiogenic) were delivered locally immediately after a contusion injury produced using the NYU impactor and a 25 mm weight-drop. Rats treated with VEGF showed significantly improved behavior up to 6 weeks after injury compared with control animals, while angiostatin treatment lead to no statistically significant changes in behavior outcome. Furthermore, VEGF-treated animals had an increased amount of spared tissue in the lesion center and a higher blood vessel density in parts of the wound area compared with controls. These effects were unlikely to be due to increased cell proliferation as determined by bromo-deoxy-uridine-labeling. Moreover, VEGF treatment led to decreased levels of apoptosis, as revealed by TUNEL assays. In situ hybridization demonstrated presence of mRNA for VEGF receptors Flt-1, fetal liver kinase-1, neuropilin-1 and -2 in several important cellular compartments of the spinal cord. The different experiments indicate that beneficial effects seen by acute VEGF delivery was attributable to protection/repair of blood vessels, decreased apoptosis and possibly also by other additional effects on glial cells or certain neuron populations.

Wilson, A. D., A. Hart, et al. (2003). "Primary sensory neuronal rescue with systemic acetyl-L-carnitine following peripheral axotomy. A dose-response analysis." Br J Plast Surg 56(8): 732-9.
 The loss of a large proportion of primary sensory neurons after peripheral nerve axotomy is well documented. As a consequence of this loss, the innervation density attained on completion of regeneration will never be normal, regardless of how well the individual surviving neurons regenerate. Acetyl-L-carnitine (ALCAR), an endogenous peptide in man, has been demonstrated to protect sensory neurons, thereby avoiding loss after peripheral nerve injury. In this study we examined the dose-response effect of ALCAR on the primary sensory neurons in the rat dorsal root ganglia (DRG) 2 weeks after sciatic nerve axotomy.Six groups of adult rats (n=5) underwent unilateral sciatic nerve axotomy, without repair, followed by 2 weeks systemic treatment with one of five doses of ALCAR (range 0.5-50 mg/kg/day), or normal saline. L4 and L5 dorsal root ganglia were then harvested bilaterally and sensory neuronal cell counts obtained using the optical disector technique. ALCAR eliminated neuronal loss at higher doses (50 and 10 mg/kg/day), while lower doses did result in loss (12% at 5 mg/kg/day, p<0.05; 19% at 1 mg/kg/day, p<0.001; 23% at 0.5 mg/kg/day, p<0.001) compared to contralateral control ganglia. Treatment with normal saline resulted in a 25% (p<0.001) loss, demonstrating no protective effect in accordance with previous studies.ALCAR preserves the sensory neuronal cell population after axotomy in a dose-responsive manner and as such, has potential for improving the clinical outcome following peripheral nerve trauma when doses in excess of 10 mg/kg/day are employed.

Wolferstan, F. (2003). "Slow neurodegeneration and transmissible spongiform encephalopathies/prion diseases. Hypothesis: a cycle involving repeated tyrosine kinase A activation could drive the development of TSEs." Med Hypotheses 60(1): 52-64.
 Neurons are specialised non-mitogenic cells. They cannot be replaced after damage, but most survive the lifetime of the individual. This is achieved by a very specialised process of repair and regeneration.During this process, a phase of degeneration in the distal end of the damaged neuron occurs in response to tyrosine kinase activation by nerve growth factor, which results in removal of neuronal detritus from within the cell membrane. As this phase is completed the activity of tyrosine kinase is modulated and the regeneration phase begins.It is postulated that normal prions play a part in the modulation of tyrosine kinase activity; that abnormal prion isoforms may be damaged in the process releasing a few fragments of prion PrP106-126 and that these stimulate release of nerve growth factor, which activates tyrosine kinase once more, setting up the vicious spiral of slow neurodegeneration found in the transmissible spongiform encephalopathies.

Wong, E. V., S. David, et al. (2003). "Inactivation of myelin-associated glycoprotein enhances optic nerve regeneration." J Neurosci 23(8): 3112-7.
 CNS regeneration in higher vertebrates is a long sought after goal in neuroscience. The lack of regeneration is attributable in part to inhibitory factors found in myelin (Caroni and Schwab, 1988a). Myelin-associated glycoprotein (MAG) is an abundant myelin protein that inhibits neurite outgrowth in vitro (McKerracher et al., 1994; Mukhopadhyay et al., 1994), but its role in regeneration remains controversial. To address this role, we performed nerve crush on embryonic day 15 chick retina-optic nerve explants and then acutely eliminated MAG function along the nerve using chromophore-assisted laser inactivation (CALI). CALI of MAG permitted significant regrowth of retinal axons past the site of lesion containing CNS myelin in contrast to various control treatments. Electron microscopy of the site of nerve crush shows abundant regenerating axons crossing the gap. When crushed optic nerve was retrogradely labeled at the nerve stump, no labeling of retinal neurons was observed. In contrast, labeling of CALI of MAG-treated crushed optic nerve showed significant retinal labeling (89 +/- 16 cells per square millimeter), a value indistinguishable from that seen with non-crushed nerve (98 +/- 13 cells per square millimeter). These findings implicate MAG as an important component of the myelin-derived inhibition of nerve regeneration. The acute loss of MAG function can promote significant axon growth across a site of CNS nerve damage.

Woodall, A. J., H. Naruo, et al. (2003). "Anesthetic treatment blocks synaptogenesis but not neuronal regeneration of cultured Lymnaea neurons." J Neurophysiol 90(4): 2232-9.
 Trauma and injury necessitate the use of various surgical interventions, yet such procedures themselves are invasive and often interrupt synaptic communications in the nervous system. Because anesthesia is required during surgery, it is important to determine whether long-term exposure of injured nervous tissue to anesthetics is detrimental to regeneration of neuronal processes and synaptic connections. In this study, using identified molluscan neurons, we provide direct evidence that the anesthetic propofol blocks cholinergic synaptic transmission between soma-soma paired Lymnaea neurons in a dose-dependent and reversible manner. These effects do not involve presynaptic secretory machinery, but rather postsynaptic acetylcholine receptors were affected by the anesthetic. Moreover, we discovered that long-term (18-24 h) anesthetic treatment of soma-soma paired neurons blocked synaptogenesis between these cells. However, after several hours of anesthetic washout, synapses developed between the neurons in a manner similar to that seen in vivo. Long-term anesthetic treatment of the identified neurons visceral dorsal 4 (VD4) and left pedal dorsal 1 (LPeD1) and the electrically coupled Pedal A cluster neurons (PeA) did not affect nerve regeneration in cell culture as the neurons continued to exhibit extensive neurite outgrowth. However, these sprouted neurons failed to develop chemical (VD4 and LPeD1) and electrical (PeA) synapses as observed in their control counterparts. After drug washout, appropriate synapses did reform between the cells, although this synaptogenesis required several days. Taken together, this study provides the first direct evidence that the clinically used anesthetic propofol does not affect nerve regeneration. However, the formation of both chemical and electrical synapses is severely compromised in the presence of this drug. This study emphasizes the importance of short-term anesthetic treatment, which may be critical for the restoration of synaptic connections between injured neurons.

Woolf, C. J. (2003). "No Nogo: now where to go?" Neuron 38(2): 153-6.
 Nogo-A, a reticulon protein expressed by oligodendrocytes, contributes to the axonal growth inhibitory action of central myelin in growth cone collapse and neurite outgrowth in vitro assays, and antibody and inhibitor studies have implicated a role for Nogo in regeneration in the adult CNS in vivo. Three independent labs have now produced Nogo knockout mice with, quite unexpectedly, three different regeneration phenotypes.

Wu, D. Y., J. Q. Zheng, et al. (2003). "PKC isozymes in the enhanced regrowth of retinal neurites after optic nerve injury." Invest Ophthalmol Vis Sci 44(6): 2783-90.
 PURPOSE: To establish an in vitro model of axonal regeneration from mammalian retinal ganglion cells and to evaluate the role of PKC isozymes in promoting such retinal axon regeneration. METHODS: Postnatal day-3 mice were subjected to optic nerve crush, and then retinal ganglion cells (RGCs) were used for culture 5 days later. RGCs were selected using anti-Thy 1.2-coated magnetic beads and plated onto a merosin substrate. Changes in axonal localization of PKC and axonal regeneration were examined in cultured RGCs by immunofluorescence. Changes in PKC isozyme mRNA levels were determined by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR). The role of PKC in RGC neurite outgrowth was examined by treatment with activators or pharmacological inhibitors of PKC activity. RESULTS: RGCs subjected to optic nerve crush injury demonstrated more rapid neurite outgrowth in vitro when compared with RGCs isolated from naive retina. The neurites of these injury-conditioned RGCs showed both an increased rate of extension and enhanced PKC localization in culture. Injury-conditioned RGCs had elevated PKC isozyme mRNA levels, which probably contributed to the increased level of PKC protein in injury-conditioned RGC axons. Pharmacological activation of PKC enhanced neurite growth, whereas inhibition of PKC suppressed neurite growth in both the conditioned and naive RGCs. CONCLUSIONS: RGCs actively respond to axonal injury by regulating expression of genes that promote neurite outgrowth. PKC-alpha and -beta isozymes are among the growth-associated proteins that are upregulated after injury. Results of pharmacological manipulation of PKC activity support the argument that increased PKC levels enhance neurite regrowth after axonal injury.

Wu, S., Y. Suzuki, et al. (2003). "Bone marrow stromal cells enhance differentiation of cocultured neurosphere cells and promote regeneration of injured spinal cord." J Neurosci Res 72(3): 343-51.
 Transplantation of bone marrow stromal cells (MSCs) has been regarded as a potential approach for promoting nerve regeneration. In the present study, we investigated the influence of MSCs on spinal cord neurosphere cells in vitro and on the regeneration of injured spinal cord in vivo by grafting. MSCs from adult rats were cocultured with fetal spinal cord-derived neurosphere cells by either cell mixing or making monolayered-feeder cultures. In the mixed cell cultures, neuroshpere cells were stimulated to develop extensive processes. In the monolayered-feeder cultures, numerous processes from neurosphere cells appeared to be attracted to MSCs. In an in vivo experiment, grafted MSCs promoted the regeneration of injured spinal cord by enhancing tissue repair of the lesion, leaving apparently smaller cavities than in controls. Although the number of grafted MSCs gradually decreased, some treated animals showed remarkable functional recovery. These results suggest that MSCs might have profound effects on the differentiation of neurosphere cells and be able to promote regeneration of the spinal cord by means of grafting.

Wu, M. M., S. W. You, et al. (2003). "Effects of inosine on axonal regeneration of axotomized retinal ganglion cells in adult rats." Neurosci Lett 341(1): 84-6.
 Damage to the central nervous system (CNS) is always followed by an irreversible axon degeneration of injured neurons. The purine nucleoside inosine has been shown to induce neurons to regenerate axons in culture and in vivo. In the present study, we investigated the in vivo effects of inosine on the axon regeneration of axotomized retinal ganglion cells (RGCs) in adult rats, using the model of peripheral nerve (PN) grafting onto the ocular stump of the transected optic nerve. Animals were allowed to survive for 4 weeks after surgery with repeated intraperitoneal injections of inosine 1 day before PN grafting till they were killed. Treatment with inosine induced a significant increase (62%) in the number of FluroGold -labeled RGCs regrowing their axons into the PN graft, when compared with the control animals. The axon outgrowth-promoting effect of inosine in adult rodents may represent a potential clinical treatment for injured or degenerated CNS.

Wu, H. H., S. Ivkovic, et al. (2003). "Autoregulation of neurogenesis by GDF11." Neuron 37(2): 197-207.
 In the olfactory epithelium (OE), generation of new neurons by neuronal progenitors is inhibited by a signal from neurons themselves. Here we provide evidence that this feedback inhibitory signal is growth and differentiation factor 11 (GDF11). Both GDF11 and its receptors are expressed by OE neurons and progenitors, and GDF11 inhibits OE neurogenesis in vitro by inducing p27(Kip1) and reversible cell cycle arrest in progenitors. Mice lacking functional GDF11 have more progenitors and neurons in the OE, whereas mice lacking follistatin, a GDF11 antagonist, show dramatically decreased neurogenesis. This negative autoregulatory action of GDF11 is strikingly like that of its homolog, GDF8/myostatin, in skeletal muscle, suggesting that similar strategies establish and maintain proper cell number during neural and muscular development.

Xiao, C. G., M. X. Du, et al. (2003). "An artificial somatic-central nervous system-autonomic reflex pathway for controllable micturition after spinal cord injury: preliminary results in 15 patients." J Urol 170(4 Pt 1): 1237-41.
 PURPOSE: Neurogenic bladder dysfunction after spinal cord injury (SCI) is a major medical and social problem for which there is no definitive solution. After the successful establishment in animals of a skin-central nervous system-bladder reflex pathway for micturition we performed this procedure on 15 patients with SCI who had 3 years of followup. MATERIALS AND METHODS: A total of 15 male volunteers with hyperreflexic neurogenic bladder and detrusor external sphincter dyssynergia (DESD) caused by complete suprasacral SCI underwent limited hemilaminectomy and ventral root (VR) micro anastomosis, usually between the L5 and S2/3 VRs. The L5 dorsal root was left intact as the trigger of micturition after axonal regeneration. Mean followup was 3 years. All patients underwent urodynamic evaluation before surgery and during followup. RESULTS: Preoperative studies in patients with complete suprasacral SCI revealed hyperreflexic neurogenic bladders and DESD with some differences in storage function during infusion cystometrograms. Of the 15 patients 10 (67%) regained satisfactory bladder control within 12 to 18 months after VR micro anastomosis. Average residual urine decreased from 332 to 31 ml and urinary infection as well as overflow incontinence disappeared. Urodynamic studies revealed a change from detrusor hyperreflexia with DESD and high detrusor pressure to almost normal storage and synergic voiding without DESD. Impaired renal function returned to normal. Two patients (13%) who required a skin stimulator to evoke voiding following the VR anastomosis had partial recovery but more than 100 ml residual urine. One patient was lost to followup and 2 had failure. CONCLUSIONS: An artificial somatic-central nervous system-autonomic reflex arc can be established surgically to provide a novel method for controlling bladder function in patients with complete suprasacral SCI who have hyperreflexic bladder and DESD. Nerve impulses delivered from the efferent neurons of a somatic reflex arc can be transferred to initiate the response of an autonomic effector.

Xu, X. J., Y. J. Piao, et al. (2003). "[Effect of human hair keratin implant on oligodendrocyte proliferation and differentiation in rats with acute spinal cord injury]." Di Yi Jun Yi Da Xue Xue Bao 23(6): 542-5.
 OBJECTIVE: To investigate the changes of oligodendrocyte proliferation and differentiation in response to implantation of degradable human hair keratin (HHK) into the spinal cord with acute injury in adult rats. METHOD: Rat models of acute spinal cord injury were established by direct impact of the exposed spinal cord with a weight-dropping device, 12 of which received immediate HHK implant into the injured spinal cord. The rats of control injury group (n=12) were subjected to the injury but did not receive subsequent implant, and those in sham operation group (n=12) only had the spinal cord exposure without injury. Another 8 rats were used as the normal control group. Samples of the spinal cord were obtained 1, 4, 12, and 26 weeks respectively after the operations and serial sections were prepared for examination with light and electron microscope. RESULTS: One week after the injury, few oligodendrocytes were observed among the large number of the infiltrating leukocytes in the injured spinal cord with HHK implants. Staining with hematoxylin eosin and Mallory's phosphotungstic acid-hematoxylin at the fourth week revealed oligodendrocyte proliferation around the HHK implant. The period from the 12th to 26th week was characterized by nerve regeneration and myelin sheath reconstruction, in the course of which empty cavity occurred within the sheath of the oligodendrocytes, and lamellar separation of the myelin sheath were observed. Phagocytized myelin sheath and axone fragment were detected in oligodendrocytes, with the newly generated oligodendrocytes scattering abound the rebuilt myelin sheath. CONCLUSION: HHK can be beneficial for the proliferation and differentiation of oligodendrocytes and myelin sheath rebuilding and repair in the course of neuronal regeneration after acute spinal cord injury.

Xu, X., W. C. Yee, et al. (2003). "Peripheral nerve regeneration with sustained release of poly(phosphoester) microencapsulated nerve growth factor within nerve guide conduits." Biomaterials 24(13): 2405-12.
 Prolonged delivery of neurotrophic proteins to the target tissue is valuable in the treatment of various disorders of the nervous system. We have tested in this study whether sustained release of nerve growth factor (NGF) within nerve guide conduits (NGCs), a device used to repair injured nerves, would augment peripheral nerve regeneration. NGF-containing polymeric microspheres fabricated from a biodegradable poly(phosphoester) (PPE) polymer were loaded into silicone or PPE conduits to provide for prolonged, site-specific delivery of NGF. The conduits were used to bridge a 10 mm gap in a rat sciatic nerve model. Three months after implantation, morphological analysis revealed higher values of fiber diameter, fiber population and fiber density and lower G-ratio at the distal end of regenerated nerve cables collected from NGF microsphere-loaded silicone conduits, as compared with those from control conduits loaded with either saline alone, BSA microspheres, or NGF protein without microencapsulation. Beneficial effects on fiber diameter, G-ratio and fiber density were also observed in the permeable PPE NGCs. Thus, the results confirm a long-term promoting effect of exogenous NGF on morphological regeneration of peripheral nerves. The tissue-engineering approach reported in this study of incorporation of a microsphere protein release system into NGCs holds potential for improved functional recovery in patients whose injured nerves are reconstructed by entubulation.

Yamaguchi, I., S. Itoh, et al. (2003). "The chitosan prepared from crab tendons: II. The chitosan/apatite composites and their application to nerve regeneration." Biomaterials 24(19): 3285-92.
 The chitosan tubes derived from crab tendons form a hollow tube structure, which is useful for nerve regeneration. However, in order to use the chitosan tubes effectively for nerve regeneration, there remain two problems to be solved. First, the mechanical strength of the tubes is quite high along the longitudinal axis, but is somewhat low for a pressure from side. Second, the chitosan tube walls swell to reduce the inner space of the tubes in vivo. These two problems limit the clinical use of the chitosan tubes. In this study, to solve the problems, apatite was made to react with the chitosan tubes to enhance the mechanical strength of the tube walls. Transmission electron microscopy showed that apatite crystals were formed in the walls of the chitosan tubes. The c-axis of the crystals aligned well in parallel with chitosan molecules. These results indicate that the apatite crystals grow in the tubes starting from the nucleation sites of the chitosan molecules, probably by forming complexes with amino groups of chitosan and calcium ions. Further, the tubes were thermally annealed at 120 degrees C to prevent from swelling, and simultaneously formed into a triangular shape to enhance the stabilization of the tube structure. By these treatments, the hollow tubes could keep their shape even in vivo after implantation. Animal tests using SD rats further showed that the chitosan tubes effectively induced the regeneration of nerve tissue, and were gradually degraded and absorbed in vivo.

Yamamoto, Y., S. Sasaki, et al. (2003). "Alternative approach using the combined technique of nerve crossover and cross-nerve grafting for reanimation of facial palsy." Microsurgery 23(3): 251-6.
 An alternative approach, using a combination of nerve crossover and cross-nerve grafting technique in a single-stage procedure, was developed for the reconstruction of reversible facial palsy. This combined technique provides some benefits such as early facial reanimation resulting from the single-stage procedure, less morbidity and sufficient innervation with an application of the end-to-side anastomosis method, and efficient neural regeneration due to coaptation of the intratemporal facial nerve. Facial nerve rehabilitation, based on double innervation by hypoglossal and contralateral healthy facial nerves, takes advantage of reliable and physiological facial reanimation.

Yamashita, T. and M. Tohyama (2003). "The p75 receptor acts as a displacement factor that releases Rho from Rho-GDI." Nat Neurosci 6(5): 461-7.
 The neurotrophin receptor p75(NTR) is involved in the regulation of axonal elongation by neurotrophins as well as several myelin components, including Nogo, myelin-associated glycoprotein (MAG) and myelin oligodendrocyte glycoprotein (OMgp). Neurotrophins stimulate neurite outgrowth by inhibiting Rho activity, whereas myelin-derived proteins activate RhoA and thereby inhibit growth. Here we show that direct interaction of the Rho GDP dissociation inhibitor (Rho-GDI) with p75(NTR) initiates the activation of RhoA, and this interaction between p75(NTR) and Rho-GDI is strengthened by MAG or Nogo. We also found that p75(NTR) facilitates the release of prenylated RhoA from Rho-GDI. The peptide ligand that is associated with the fifth alpha helix of p75(NTR) inhibits the interaction between Rho-GDI and p75(NTR), thus silencing the action mediated by p75(NTR). This peptide has potential as a therapeutic agent against the inhibitory cues that block regeneration in the central nervous system.

Yang, R. K., J. B. Lowe, 3rd, et al. (2003). "Dose-dependent effects of FK506 on neuroregeneration in a rat model." Plast Reconstr Surg 112(7): 1832-40.
 This study explored the effects of different doses of FK506 on peripheral nerve regeneration, to determine whether neuroregeneration could be enhanced without the toxicity of systemic immunosuppression. In the first part of the study, subimmunosuppressive doses of FK506 were determined by examining skin allograft survival in a rat model. Full-thickness skin grafts (2 cm2) from Wistar rats were grafted to recipient Lewis rats. The procedure was performed for six groups (n = 6). The control group received no FK506, and the other five groups received daily doses of FK506 of 0.125, 0.25, 0.5, 1.0, or 2.0 mg/kg. Animals that received 2.0 mg/kg FK506 per day exhibited complete skin graft take, whereas all other groups demonstrated complete rejection. After determination of the immunosuppressive dose of FK506, the neuroregenerative effects of different doses of FK506 were explored by assessing nerve regeneration in 80 rats after tibial nerve transection and repair. The control group received no FK506, whereas the other four groups were given daily doses of FK506 of 0.25, 0.5, 1.0, or 2.0 mg/kg. Rats were euthanized at three time points (25, 30, and 35 days), to fully investigate the effects of different FK506 dosing regimens on neuroregeneration. Histomorphometric analyses performed on postoperative days 30 and 35 demonstrated statistically significant improvements in neuroregeneration with subimmunosuppressive FK506 doses of 0.5 and 1.0 mg/kg per day. Therefore, the study demonstrated that neuroregeneration was enhanced at low doses of FK506 that were not sufficient to prevent skin allograft rejection.

Yang, H., C. Liang, et al. (2003). "[Proteomics analysis and biochemical activity detection on facial nerve regeneration fluid]." Lin Chuang Er Bi Yan Hou Ke Za Zhi 17(1): 37-9.
 OBJECTIVE: To explore the protein components and the biological neutrional activity upon motor neuron, so that take a general, basic study on the nerve regeneration microenvironment which be important to facial nerve regeneration. METHOD: The animal model of rabbit facial regeneration microenvironment is established. Advanced technology of two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) was used to study on the structural proteins in the regeneration fluid, as well as the motor neuron culture was used to test the biological activity in it. RESULT: 850 +/- 78 protein spots were detected in the regeneration fluid. The body area, neurite length and OD value of experimental group are higher than that of control group. CONCLUSION: We got the protein image of facial nerve regeneration fluid by 2-D PAGE, and proved that many kinds and number of proteins in the regeneration fluid have neutrional activity on motor neuron.

Yang, H., C. Yang, et al. (2003). "[Using two-dimensional gel electrophoresis to analyze the regeneration microenvironment of facial nerve and its response to acupuncture]." Sichuan Da Xue Xue Bao Yi Xue Ban 34(2): 242-4.
 OBJECTIVE: To investigate the distribution of proteins in the regeneration microenvironment of facial nerve and its response to acupuncture. METHODS: We created an animal model using rabbit to imitate the regeneration microenvironment of facial nerve and analyzed the proteins in the microenvironment through the two-dimensional gel electrophoresis. RESULTS: We found that most proteins in the regeneration fluid were under relative molecular mass 80 x 10(3) and in the range of pI 4-7. An average of 800 +/- 53 protein spots was found in the control group compared to the 850 +/- 78 spots in the acupuncture group. In the matching spots, more than ten proteins were different between the two groups. CONCLUSION: There is no evidence to show that acupuncture has a positive effect on the regeneration microenvironment of facial nerve.

Yasuda, H., M. Terada, et al. (2003). "Diabetic neuropathy and nerve regeneration." Prog Neurobiol 69(4): 229-85.
 Diabetic neuropathy is the most common peripheral neuropathy in western countries. Although every effort has been made to clarify the pathogenic mechanism of diabetic neuropathy, thereby devising its ideal therapeutic drugs, neither convinced hypotheses nor unequivocally effective drugs have been established. In view of the pathologic basis for the treatment of diabetic neuropathy, it is important to enhance nerve regeneration as well as prevent nerve degeneration. Nerve regeneration or sprouting in diabetes may occur not only in the nerve trunk but also in the dermis and around dorsal root ganglion neurons, thereby being implicated in the generation of pain sensation. Thus, inadequate nerve regeneration unequivocally contributes to the pathophysiologic mechanism of diabetic neuropathy. In this context, the research on nerve regeneration in diabetes should be more accelerated. Indeed, nerve regenerative capacity has been shown to be decreased in diabetic patients as well as in diabetic animals. Disturbed nerve regeneration in diabetes has been ascribed at least in part to all or some of decreased levels of neurotrophic factors, decreased expression of their receptors, altered cellular signal pathways and/or abnormal expression of cell adhesion molecules, although the mechanisms of their changes remain almost unclear. In addition to their steady-state changes in diabetes, nerve injury induces injury-specific changes in individual neurotrophic factors, their receptors and their intracellular signal pathways, which are closely linked with altered neuronal function, varying from neuronal survival and neurite extension/nerve regeneration to apoptosis. Although it is essential to clarify those changes for understanding the mechanism of disturbed nerve regeneration in diabetes, very few data are now available. Rationally accepted replacement therapy with neurotrophic factors has not provided any success in treating diabetic neuropathy. Aside from adverse effects of those factors, more rigorous consideration for their delivery system may be needed for any possible success. Although conventional therapeutic drugs like aldose reductase (AR) inhibitors and vasodilators have been shown to enhance nerve regeneration, their efficacy should be strictly evaluated with respect to nerve regenerative capacity. For this purpose, especially clinically, skin biopsy, by which cutaneous nerve pathology including nerve regeneration can be morphometrically evaluated, might be a safe and useful examination.

Yasumatsu, K., H. Katsukawa, et al. (2003). "Recovery of amiloride-sensitive neural coding during regeneration of the gustatory nerve: behavioral-neural correlation of salt taste discrimination." J Neurosci 23(10): 4362-8.
 The chorda tympani (CT) nerve innervating the anterior tongue contains two types of NaCl-responsive fibers: one, the N-type, receives input from receptor cells, the NaCl responses of which are strongly inhibited by amiloride, whereas the other, the E-type, receives input from cells poorly sensitive or insensitive to amiloride. To investigate the formation of this differentially responsive neural system, we crushed the mouse CT nerve and examined the subsequent recovery of NaCl responses and amiloride sensitivity of the regenerated nerve and behavioral discrimination between NaCl and KCl. At 2 weeks after the nerve crush, no significant response of the nerve to chemical stimuli was observed. At 3 weeks, responses to salts gradually reappeared. In this period, almost all single fibers responding to NaCl were insensitive to amiloride (E-type). At 4 weeks, some of the single fibers showed amiloride sensitivity (N-type) and behavioral discrimination between NaCl and KCl reappeared. After >or=5 weeks, the number of N-type fibers had reached the control level and became approximately equal to that of E-type fibers. During the course of recovery, N-type and E-type fibers were clearly distinguishable on the basis of their amiloride sensitivities, their KCl/NaCl response ratios, and their concentration-response relationships to NaCl. These results suggest that two salt-responsive systems are independently reformed after the nerve crush. The selective synapse reformation may account for recovery of behavioral discrimination between NaCl and KCl after taste nerve crush and regeneration. It may also explain stable sensory coding for taste quality during the continuous turnover of receptor cells in the healthy animal.

Yick, L. W., P. T. Cheung, et al. (2003). "Axonal regeneration of Clarke's neurons beyond the spinal cord injury scar after treatment with chondroitinase ABC." Exp Neurol 182(1): 160-8.
 We have previously demonstrated that enzymatic digestion of chondroitin sulfate proteoglycan (CSPG) at the scar promotes the axonal regrowth of Clarke's nucleus (CN) neurons into an implanted peripheral nerve graft after hemisection of the spinal cord. The present study examined whether degradation of CSPG using chondroitinase ABC promoted the regeneration of CN neurons through the scar into the rostral spinal cord in neonatal and adult rats. Following hemisection of the spinal cord at T11, either vehicle or chondroitinase ABC was applied onto the lesion site. The postoperative survival periods were 2 and 4 weeks. The regenerated CN neurons were retrogradely labeled by Fluoro-Gold injected at spinal cord level C7. In the sham group, there was no regeneration of injured CN neurons in both neonatal and adult rats. Treatment with 2.5 unit/ml chondroitinase ABC in neonates resulted in 11.8 and 8.3% of the injured CN neurons regenerated into the rostral spinal cord at 2 and 4 weeks, respectively. In adults, 9.4 and 12.3%, at 2 and 4 weeks, respectively, of the injured CN neurons regenerated their axons to the rostral spinal cord. The immunoreactivity for the carbohydrate epitope of CSPG was dramatically decreased around the lesion site after treatment with chondroitinase ABC compared to sham control in both neonatal and adult animals. Our results show that axonal regeneration in the spinal cord can be promoted by degradation of CSPG with chondroitinase ABC. This result further suggests that CSPG is inhibitory to the regeneration of neurons in the spinal cord after traumatic injury.

Yin, Y., Q. Cui, et al. (2003). "Macrophage-derived factors stimulate optic nerve regeneration." J Neurosci 23(6): 2284-93.
 After optic nerve injury in mature mammals, retinal ganglion cells (RGCs) are normally unable to regenerate their axons and undergo delayed apoptosis. However, if the lens is damaged at the time of nerve injury, many RGCs survive axotomy and regenerate their axons into the distal optic nerve. Lens injury induces macrophage activation, and we show here that factors secreted by macrophages stimulate RGCs to regenerate their axons. When macrophages were activated by intravitreal injections of Zymosan, a yeast cell wall preparation, the number of RGC axons regenerating into the distal optic nerve was even greater than after lens injury. These effects were further enhanced if Zymosan was injected 3 d after nerve crush. In a grafting paradigm, intravitreal Zymosan increased the number of RGCs that regenerated their axons through a 1.5 cm peripheral nerve graft twofold relative to uninjected controls and threefold if injections were delayed 3 d. In cell culture, media conditioned by activated macrophages stimulated adult rat RGCs to regenerate their axons; this effect was potentiated by a low molecular weight factor that is constitutively present in the vitreous humor. After gel-filtration chromatography, macrophage-derived proteins > or =30 kDa were found to be toxic to RGCs, whereas proteins <30 kDa reversed this toxicity and promoted axon regeneration. The protein(s) that stimulated axon growth is distinct from identified polypeptide trophic factors that were tested. Thus, macrophages produce proteins with both positive and negative effects on RGCs, and the effects of macrophages can be optimized by the timing of their activation.

Yiou, R., J. J. Yoo, et al. (2003). "Restoration of functional motor units in a rat model of sphincter injury by muscle precursor cell autografts." Transplantation 76(7): 1053-60.
 BACKGROUND: Urinary incontinence is a debilitating condition that affects primarily elderly individuals. One major mechanism results from chronic denervation of the striated urethral sphincter with associated fibrosis. The authors investigated the fate of muscle precursor cells (MPC) injected into a model of striated urethral sphincter injury that reproduces the histopathologic changes of sphincter insufficiency. METHODS: The striated urethral sphincter of older male rats was damaged by electrocoagulation. MPC were isolated from limb myofiber explants, infected with an adenovirus carrying the transgene encoding beta-galactosidase, and injected into the sphincter of the same animal 37 days after injury. Animals were killed 5 and 30 days after injection for assessment of sphincter function and the formation of motor units. RESULTS: Electrocoagulation resulted in an irreversible destruction of both sphincteric myofibers and nerve endings, with a functional incapacity of the damaged sphincter to sustain an increase in bladder pressure; atrophy and fibrosis developed after 1 month. Injection of MPC resulted in the formation of beta-galactosidase-expressing myotubes in the sphincter that persisted beyond 30 days. The regenerated myotubes carried acetylcholine receptors associated with a nerve ending and were thus considered to form anatomic motor units. Urodynamic studies confirmed the restoration of 41% of sphincter function 1 month after MPC injection. CONCLUSIONS: The authors showed that MPC isolated from limb muscles of an older animal can recapitulate a myogenic program when injected into an irreversibly injured sphincter. The maturation of MPC activates nerve regeneration and restores functional motor units.

Yip, S., K. S. Aboody, et al. (2003). "Neural stem cell biology may be well suited for improving brain tumor therapies." Cancer J 9(3): 189-204.
 Neural stem cells (NSCs) are capable of tremendous migratory potential to areas of pathology in the central nervous system. When implanted into a diseased or injured nervous system, NSCs can travel through great distances to and engraft within areas of discrete as well as diffuse abnormalities. Engraftment is often followed by integration into the local neural milieu, accompanied by stable gene expression from the NSCs. In addition, the pluripotency of NSCs endows them with the capability to replace diseased tissues in an appropriate manner. Recent evidence has also suggested that engrafted exogenous NSCs may have effects on the surrounding microenvironment, such as promoting protection and/or regeneration of host neural pathways. These characteristics of NSCs would seem to make them ideal agents for the treatment of various central nervous system pathologies, especially brain tumors. Brain tumors are generally difficult to treat because of the unique location of the lesions. In primary gliomas, the extensive infiltrative nature of the tumor cells presents a challenge for their effective and total eradication, hence the high rate of treatment failure and disease recurrence. In addition, normal brain structures are distorted and are often destroyed by the growing neoplasm. Even with effective therapy to surgically resect and destroy the neoplastic tissues, the brain is still injured, which often leaves the patient in a debilitated state. The unique ability of NSCs to "home in" on tumor cells followed by the delivery of a desired gene product makes the NSC a very promising agent in brain tumor therapy. Cytolytic viruses and genes coding for anti-tumor cytokines, pro-drug converting enzymes, and various neurotrophic factors have all been engineered into engraftable NSCs for delivery to tumors. When they are specially tagged, such injected NSCs can be visualized with the use of novel imaging techniques and tracked in vivo within living animals over real time. If the NSCs were also capable of participating in the subsequent repair and regeneration of the tumor-afflicted brain-at present a potential but as-yet-unproven aspect of this intervention-then its role in abetting anti-tumor therapy would be complete. It is important to emphasize, however, that the use of NSCs is adjunctive and is not a replacement for other therapies that should be used in parallel.

Yoon, S., H. W. Lee, et al. (2003). "Upregulation of TrkA neurotrophin receptor expression in the thymic subcapsular, paraseptal, perivascular, and cortical epithelial cells during thymus regeneration." Histochem Cell Biol 119(1): 55-68.
 Neuroimmune networks in the thymic microenvironment are thought to be involved in the regulation of T cell development. Here, we report upon an examination of the expression of the TrkA neurotrophin receptor, the high affinity receptor for nerve growth factor, during regeneration following acute involution induced by cyclophosphamide in the rat thymus. Light and electron microscopic immunocytochemistry demonstrated enhanced expression of the TrkA receptor in the subcapsular, paraseptal, perivascular, and cortical epithelial cells during thymus regeneration. In addition, various morphological alterations, suggestive of a hyperfunctional and dynamic state, of the subcapsular, paraseptal, and perivascular epithelial cells were also observed. The presence of TrkA protein in extracts from the control and regenerating rat thymus was confirmed by western blot. Furthermore, RT-PCR analysis supported these results by demonstrating that thymic extracts contain TrkA mRNA at higher levels during thymus regeneration. Thus, our results suggest that the TrkA receptor located on the thymic subcapsular, paraseptal, perivascular, and cortical epithelial cells could play a role in the development of new T cells to replace T cells damaged during thymus regeneration.

Yoshii, S., M. Oka, et al. (2003). "Bridging a 30-mm nerve defect using collagen filaments." J Biomed Mater Res 67A(2): 467-74.
 This article describes a 30-mm regeneration of severed peripheral nerve axons along collagen filaments. Two thousand or 4000 31-mm-long collagen filaments were grafted to bridge a 30-mm defect of the rat sciatic nerve. A collagen tube was grafted as a control. The mean number and mean fiber diameter of regenerated myelinated axons were 330 +/- 227 and 2.7 +/- 0.9 microm in the distal end of the 2000 collagen-filaments nerve guide, and 564 +/- 275 and 2.5 +/- 1.1 microm in the distal end of the 4000 collagen-filaments nerve guide at 12 weeks postoperatively, whereas in the distal end of the collagen tube, no regenerated axon was found. These results suggest that the collagen filaments guide axons of the rat's sciatic nerve to regenerate for 30 mm and act as a scaffold for axonal regeneration. Thirty-millimeter nerve regeneration of a 1-mm-diameter rat sciatic nerve by an artificial nerve guarantees a clinical application of the implant which should be very important for patients and surgeons.

Yoshii, S., M. Oka, et al. (2003). "Bridging a spinal cord defect using collagen filament." Spine 28(20): 2346-51.
 SUMMARY: STUDY DESIGN A rat model of spinal cord defect was designed to evaluate the effect of collagen filament implant on nerve regeneration in the spinal cord defect.OBJECTIVES To bridge a spinal cord defect and restore the function in adult mammals.SUMMARY OF BACKGROUND DATA Resection of the spinal cord in mammals is always followed by motor paralysis and loss of voluntary function below the lesion. Partial success in bridging the ends of the spinal cord after complete resection was reported. However, restoration of function has not been reported in adult mammalian.MATERIALS AND METHODS Four thousand collagen filaments 5-mm-long were grafted to bridge a 5-mm defect of rat spinal cord. Controls had their spinal cord defect left ungrafted after resection. At 1-week intervals, animals were evaluated functionally. After 4 and 12 weeks, animals were evaluated histologically. After 12 weeks, animals were evaluated electrophysiologically.RESULTS The severed spinal cord axons regenerated along the collagen filament implant crossing the proximal and distal spinal cord implant interfaces at 4 weeks after surgery. The rats with collagen filament grafts could walk, run, and climb with hind forelimb coordination at 12 weeks after surgery. Sensory-evoked potential waveform was found in the rats with collagen filament at 12 weeks after surgery.CONCLUSIONS The collagen filaments support the axonal regeneration of the transected spinal cord and the restoration of function.

You, S. W., B. Y. Chen, et al. (2003). "Spontaneous recovery of locomotion induced by remaining fibers after spinal cord transection in adult rats." Restor Neurol Neurosci 21(1-2): 39-45.
 PURPOSE: A major issue in analysis of experimental results after spinal cord injury is spontaneous functional recovery induced by remaining nerve fibers. The authors investigated the relationship between the degree of locomotor recovery and the percentage and location of the fibers that spared spinal cord transection. METHODS: The spinal cords of 12 adult rats were transected at T9 with a razor blade, which often resulted in sparing of nerve fibers in the ventral spinal cord. The incompletely-transected animals were used to study the degree of spontaneous recovery of hindlimb locomotion, evaluated with the BBB rating scale, in correlation to the extent and location of the remaining fibers. RESULTS: Incomplete transection was found in the ventral spinal cord in 42% of the animals. The degree of locomotor recovery was highly correlated with the percentage of the remaining fibers in the ventral and ventrolateral funiculi. In one of the rats, 4.82% of remaining fibers in unilateral ventrolateral funiculus were able to sustain a certain recovery of locomotion. CONCLUSIONS: Less than 5% of remaining ventrolateral white matter is sufficient for an unequivocal motor recovery after incomplete spinal cord injury. Therefore, for studies with spinal cord transection, the completeness of sectioning should be carefully checked before any conclusion can be reached. The fact that the degree of locomotor recovery is correlated with the percentage of remaining fibers in the ventrolateral spinal cord, exclusive of most of the descending motor tracts, may imply an essential role of propriospinal connections in the initiation of spontaneous locomotor recovery.

Yu, X., L. Xu, et al. (2003). "Effect of spinal cord injury on urinary bladder spinal neural pathway: a retrograde transneuronal tracing study with pseudorabies virus." Urology 62(4): 755-9.
 OBJECTIVES: To determinate the effect of acute and chronic spinal cord injury (SCI) resulting from thoracic cord transection on the urinary bladder spinal neural pathway. METHODS: Seventy-six adult Sprague-Dawley rats were randomly divided into four groups, non-SCI (normal rats undergoing no surgical procedure except pseudorabies virus [PRV] injection), SCI(b) (SCI and PRV injected immediately after SCI), SCI(c) (SCI and PRV injected at 3 weeks after SCI), and SCI(d) (SCI and PRV injected at 3 months after SCI). Transcardiac perfusion fixation was done at appropriate survival periods after PRV injection into the bladder wall tissue. Sections of the dorsal root ganglion, spinal cord, and brain were processed for visualization of the virus by the streptavidin-peroxidase immunohistochemical procedure. RESULTS: The bladder weight of the non-SCI, SCI(b), SCI(c), SCI(d) rats was 144 +/- 9 mg, 142 +/- 8 mg, 486 +/- 51 mg, and 656 +/- 69 mg, respectively. The time-ordered flow charts of PRV tracing were similar in the non-SCI and SCI rats. The cross-sectional area of the labeled dorsal root ganglion cell profiles increased significantly after SCI (P <0.001): 593 +/- 40 microm2, 588 +/- 39 microm2, 815 +/- 53 microm2, and 902 +/- 57 microm2 in the non-SCI, SCI(b), SCI(c), SCI(d) rats, respectively. The number of labeled cells in the dorsal horn in the L6 and S1 segments 3 days after PRV injection markedly increased in chronic SCI rats, as did the number of labeled motor neurons 4 days after injection. CONCLUSIONS: Acute and chronic SCI have no effect on the process of virus transneuronal transport below the level of the lesion. Subsequent to chronic SCI, reorganization of the micturition reflex pathways may occur.

Yu, X. and R. V. Bellamkonda (2003). "Tissue-engineered scaffolds are effective alternatives to autografts for bridging peripheral nerve gaps." Tissue Eng 9(3): 421-30.
 The use of autografts for "bridging" peripheral nerve gaps is limited by lack of suitable donor nerve grafts. Using a tissue-engineering approach, we have designed a three-dimensional scaffold that presents laminin 1 (LN-1) and nerve growth factor (NGF) in vivo. Semipermeable polysulfone tubes were used as carriers to introduce the tissue-engineered scaffolds to a 10-mm sciatic nerve gap in adult rats. Two months after implantation, the gross morphology of the regenerated nerve, the success rate of regeneration, and the total number and density of myelinated axons in the tissue-engineered scaffolds matched that observed in autografts. LN-1- and NGF-containing scaffolds performed comparably to autografts when functional measures that include the relative gastrocnemius muscle weight and the sciatic functional index were quantified. Our results demonstrate that tissue-engineered scaffolds match the performance of autografts in an in vivo model of peripheral nerve regeneration, raising the possibility of the scaffolds being used clinically instead of scarce autografts.

Zador, E. and F. Wuytack (2003). "Expression of SERCA2a is independent of innervation in regenerating soleus muscle." Am J Physiol Cell Physiol 285(4): C853-61.
 The speed of contraction of a skeletal muscle largely depends on the myosin heavy chain isoforms (MyHC), whereas the relaxation is initiated and maintained by the sarcoplasmic reticulum Ca2+-ATPases (SERCA). The expression of the slow muscle-type myosin heavy chain I (MyHCI) is entirely dependent on innervation, but, as we show here, innervation is not required for the expression of the slow-type sarcoplasmic reticulum Ca2+-ATPase (SERCA2a) in regenerating soleus muscles of the rat, although it can play a modulator role. Remarkably, the SERCA2a level is even higher in denervated than in innervated regenerating soleus muscles on day 7 when innervation is expected to resume. Later, the level of SERCA2a protein declines in denervated regenerated muscles but it remains expressed, whereas the corresponding mRNA level is still increasing. SERCA1 (i.e., the fast muscle-type isoform) expression shows only minor changes in denervated regenerating soleus muscles compared with innervated regenerating controls. When the soleus nerve was transected instead of the sciatic nerve, SERCA2a and MyHCI expressions were found to be even more uncoupled because the MyHCI nearly completely disappeared, whereas the SERCA2a mRNA and protein levels decreased much less. The transfection of regenerating muscles with constitutively active mutants of the Ras oncogene, known to mimic the effect of innervation on the expression of MyHCI, did not affect SERCA2a expression. These results demonstrate that the regulation of SERCA2a expression is clearly distinct from that of the slow myosin in the regenerating soleus muscle and that SERCA2a expression is modulated by neuronal activity but is not entirely dependent on it.

Zamboni, W. A. (2003). "What's new in plastic and maxillofacial surgery." J Am Coll Surg 196(3): 453-62.
 
Zalish, M. and V. Lavie (2003). "Dexanabinol (HU-211) has a beneficial effect on axonal sprouting and survival after rat optic nerve crush injury." Vision Res 43(3): 237-42.
 Dexanabinol (HU-211) is a synthetic non-psychotropic cannabinoid and a non-competitive NMDA-receptor antagonist. The beneficial effect of dexanabinol on prevention of degeneration and promotion of regeneration was studied on the crush-injured rat optic nerve model. Sprague-Dawley rats were subjected to a calibrated crush injury of the optic nerve and treated with a single intraperitoneal injection of dexanabinol (7 mg/kg), its vehicle only or were untreated. Transmission electron microscopic analysis of the excised optic nerves was performed after 30 days. In the dexanabinol treated rats, the site of injury was traversed by unmyelinated and thinly myelinated axons, possibly indicative of regenerative growth. No such growth was detectable in the controls. Viable axons were found 0.5 mm distal to the site of injury in 6 of 8 dexanabinol treated rats, but in only 1 of 10 rats in the control groups.These results have clinical implications for the prevention of secondary degeneration and promotion of regeneration after injuries to the central nervous system.

Zhang, X. F., A. W. Schaefer, et al. (2003). "Rho-dependent contractile responses in the neuronal growth cone are independent of classical peripheral retrograde actin flow." Neuron 40(5): 931-44.
 Rho family GTPases have been implicated in neuronal growth cone guidance; however, the underlying cytoskeletal mechanisms are unclear. We have used multimode fluorescent speckle microscopy (FSM) to directly address this problem. We report that actin arcs that form in the transition zone are incorporated into central actin bundles in the C domain. These actin structures are Rho/Rho Kinase (ROCK) effectors. Specifically, LPA mediates growth cone retraction by ROCK-dependent increases in actin arc and central actin bundle contractility and stability. In addition, these treatments had marked effects on MT organization as a consequence of strong MT-actin arc interactions. In contrast, LPA or constitutively active Rho had no effect on P domain retrograde actin flow or filopodium bundle number. This study reveals a novel mechanism for domain-specific spatial control of actin-based motility in the growth cone with implications for understanding chemorepellant growth cone responses and nerve regeneration.

Zhang, J., T. M. Oswald, et al. (2003). "Enhancement of rat sciatic nerve regeneration by fibronectin and laminin through a silicone chamber." J Reconstr Microsurg 19(7): 467-72.
 In this study, the authors examined the effects of fibronectin and laminin on sciatic nerve regeneration in rats. Sixty-eight Sprague-Dawley rats underwent bilateral sciatic nerve transections and silicone tubulizations, with a 10-mm gap between the proximal and distal nerve stumps. Thirty rats (n=30) received 10 microg of fibronectin injection into the right sciatic nerve chamber, while saline was injected into the left nerve chamber, serving as the control. Another 30 rats (n=30) were given 6 microg of laminin injection into the right nerve chambers and saline into the left chambers. At 1, 3, and 4 months postoperatively, electrophysiologic and histologic examinations, including nerve morphometry, were performed. Eight additional rats, receiving fibronectin (n=4) and laminin (n=4) injections, were used for horseradish peroxidase (HRP) tracing at 3 months postoperatively. Results from the study showed that fibronectin- and laminin-treated groups had significantly higher motor nerve conduction velocity and evoked muscle action potential amplitude of the anterior tibial muscle than the control group (p<0.01). Nerve diameter and the number of myelinated axons from the groups receiving fibronectin and laminin applications were greater than the controls (p<0.01). Also, a greater number of HRP-labeled motor neurons were found in the ventral horns and dorsal root ganglia of the fibronectin- and laminin-treatment groups compared to the controls. The authors conclude that local applications of fibronectin and laminin into the nerve chambers can significantly improve axonal regeneration and maturation of injured rat sciatic nerves.

Zhang, J., C. Geula, et al. (2003). "Neurotrophins regulate proliferation and survival of two microglial cell lines in vitro." Exp Neurol 183(2): 469-81.
 Microglia are thought to play a key role in the development and regeneration of the central nervous system although the mechanisms regulating their presence and activity are not fully understood. Substantial evidence suggests that members of the neurotrophin family such as nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin-3 and -4 (NT-3/4) have a dramatic effect on both neurons and perineuronal cells. This study employed two murine microglial lines, BV-2 and N9, to examine the action of these neurotrophins on the mitotic activity and survival of microglia in vitro. Neurotrophins were incorporated into the media at the time of plating and cell number and levels of mitochondrial dehydrogenase activity (MTT) were determined at various time points in vitro. NGF increased cell number and MTT levels of both cell lines in a dose-dependent manner. BV-2 was more sensitive to NGF than N9. Similar responses were elicited by BDNF, although the sensitivity of each cell line was different than that found for NGF. NT-3 and NT-4 had no effect on cell proliferation. However, NT-4 had an effect on the survival of BV-2 and N9 cells. The response of these cells to neurotrophins was blocked by K252a, a tyrosine kinase inhibitor, suggesting that actions of neurotrophins were mediated by high-affinity tyrosine kinase receptors (Trk). Immunolocalization studies revealed positive Trk (pan) reactivity in the above cell lines and in primary microglia, but an absence of the low-affinity p75 neurotrophin receptor. Western blot analysis supported the above observations. These studies suggest that in addition to their neurotrophic actions, NGF and BDNF may also regulate microglial dynamics, thereby influencing the surrounding milieu during neuronal regeneration.

Zhang, F., P. Ferretti, et al. (2003). "Recruitment of postmitotic neurons into the regenerating spinal cord of urodeles." Dev Dyn 226(2): 341-8.
 By using fluorescent tracers, we have investigated the origin of the cells that form the regenerating spinal cord after tail amputation in urodele amphibians. We show that spinal cord cells immediately adjacent to the amputation plane die and are removed by phagocytic cells. Spinal cells just anterior to these dying cells are destined to make the majority of the regenerating cord. The largest contribution is likely to come from the radial ependymal cells, but we also demonstrate that postmitotic neurons in this location can translocate into the regenerating cord. These neurons integrate into the regenerate structure and survive for at least 4 weeks. We find no evidence that these translocated neurons dedifferentiate and divide during this regeneration process. We discuss the possibility that these neurons survive long term in the regenerate cord and become part of the functional neuronal circuitry.

Zhao, Y. L., K. Takagawa, et al. (2003). "Active Src expression is induced after rat peripheral nerve injury." Glia 42(2): 184-93.
 The non-receptor-type Src tyrosine kinases are key components of intracellular signal transduction that are expressed at high levels in the nervous system. To improve understanding of the cascades of molecular events underlying peripheral nerve regeneration, we analyzed active Src expression in the crushed or cut rat sciatic nerves using a monoclonal antibody (clone 28) that recognizes the active form of Src tyrosine kinases, including c-Src and c-Fyn. Western blots showed that active Src expressed in the normal sciatic nerve transiently increased up to threefolds after both types of injury. Immunohistochemistry using clone 28 showed that axonal components are the primary sites of active Src expression in the normal sciatic nerve. Soon after both types of injury, active Src was abundantly expressed in Schwann cells of the segments distal to the injury site. The expression of active Src in the cells decreased with restoration of the axon-Schwann cell relationship and eventually became depleted to very low levels after crushing, but was sustained at high levels in the cut model until the end of the experiment. Regenerated axons consistently expressed active Src throughout nerve regeneration and these eventually became the major sites of active Src expression in the crushed nerve. Among the Src tyrosine kinases, active c-Src selectively increased after crushing according to immunoprecipitation and immunoblotting analyses. Due to its potent biological activity, the increased amounts of the active form of Src probably enhance axonal regrowth, the Schwann cell response, and axon-Schwann cell contact for peripheral nerve regeneration.

Zharkovsky, T., A. Kaasik, et al. (2003). "Neurodegeneration and production of the new cells in the dentate gyrus of juvenile rat hippocampus after a single administration of ethanol." Brain Res 978(1-2): 115-23.
 Administration of ethanol during brain development induces widespread neuronal loss in various structures of the brain. Here, we show that a single administration of ethanol given during the early postnatal period can induce not only neuronal death, but also an increase in proliferation of the progenitor cells in the dentate gyrus of hippocampal formation in rats. Ethanol (1.5 or 3 g/kg, i.p.) administered to 10-day-old rats induced massive neuronal degeneration as evidenced by TUNEL assay in the dentate gyrus. The neuronal death induced by a high dose of ethanol (3 g/kg) was accompanied by an enhanced proliferation of the progenitor cells labeled by bromodeoxyuridine (BrdU, 50 mg/kg, i.p.) in dentate gyrus. One and 3 weeks following ethanol or saline administration, ethanol-treated rats still had significantly more BrdU-labeled cells than control animals. In ethanol-treated rats, a higher proportion of newly born cells acquired the phenotype of immature postmitotic neurons whereas the final differentiation into calbindin-expressing granule cells remained unchanged. The proportion of astroglial cells was also increased in ethanol-treated rats. Thus, ethanol given in high doses not only induces neurodegeneration but also initiates the process of neuro- and gliogenesis, which might be responsible for the neuronal and glial reorganization of the dentate gyrus.

Zhelyaznik, N., K. Schrage, et al. (2003). "Activation of retinoic acid signalling after sciatic nerve injury: up-regulation of cellular retinoid binding proteins." Eur J Neurosci 18(5): 1033-40.
 In mammalian peripheral nerves a crush lesion causes interactions between injured neurons, Schwann cells and haematogenous macrophages that can lead to successful axonal regeneration. We suggest that the transcriptional activator retinoic acid (RA), takes part in gene regulation after peripheral nerve injury and that RA signalling is activated via the cellular retinoic acid binding protein (CRABP)-II and cellular retinol binding protein (CRBP)-I. With RT-PCR and immunoblotting all necessary components of the RA signalling pathway were detected in the sciatic nerve of adult rats. These are retinoic acid receptors, retinoid X receptors, the retinoic acid synthesizing enzymes RALDH-1, RALDH-2, and RALDH-3, in addition, the cellular retinoid binding proteins CRBP-I, CRABP-I and CRABP-II. Enzyme activity of RALDH-2 was detectable in the nerve, and using a transgenic reporter mouse we found local activation of RA responsive elements in the regenerating nerve. Sciatic nerve crush as well as transection resulted in a more than 10-fold up-regulation of CRBP-I, which is thought to facilitate the synthesis of RA. Both kinds of injury also caused a 15-fold increase in transcript and protein concentration of CRABP-II, a possible mediator of RA transfer to its nuclear receptors.

Zheng, B., C. Ho, et al. (2003). "Lack of enhanced spinal regeneration in Nogo-deficient mice." Neuron 38(2): 213-24.
 The failure of regeneration of severed axons in the adult mammalian central nervous system is thought to be due partly to the presence of endogenous inhibitors of axon regeneration. The nogo gene encodes three proteins (Nogo-A, -B, and -C) that have been proposed to contribute to this inhibition. To determine whether deletion of nogo enhances regenerative ability, we generated two lines of mutant mice, one lacking Nogo-A and -B but not -C (Nogo-A/B mutant), and one deficient in all three isoforms (Nogo-A/B/C mutant). Although Nogo-A/B-deficient myelin has reduced inhibitory activity in a neurite outgrowth assay in vitro, tracing of corticospinal tract fibers after dorsal hemisection of the spinal cord did not reveal an obvious increase in regeneration or sprouting of these fibers in either mouse line, suggesting that elimination of Nogo alone is not sufficient to induce extensive axon regeneration.

Zhou, L., Z. Shao, et al. (2003). "Cryoanalgesia: electrophysiology at different temperatures." Cryobiology 46(1): 26-32.
 Somatosensory evoked potentials (SEP) and sensory conduction velocity (SCV) were measured in rabbit sciatic nerves following graded cold lesioning. The SEP disappeared when injury was induced at temperatures below -60 degrees C, but returned on day 41+/-4 (mean+/-SD). SEP returned on day 56+/-11 days when the lesion was induced at 100 to -180 degrees C. The SEP latency was prolonged after creating lesions at -100 to -180 degrees C, compared with both the sham operated and the -20 degrees C groups. These experiments suggest the cryolesions produced at temperatures between -60 and -100 degrees C are most suitable for altering the electrophysiological conduction of the nerve, and may result in suitable post-operative analgesia.

Zhou, L., B. J. Baumgartner, et al. (2003). "Neurotrophin-3 expressed in situ induces axonal plasticity in the adult injured spinal cord." J Neurosci 23(4): 1424-31.
 The mammalian CNS lacks the ability to effectively compensate for injury by the regeneration of damaged axons or axonal plasticity of intact axons. However, reports suggest that molecular or cellular manipulations can induce compensatory processes that could support regeneration or plasticity after trauma. We tested whether local, sustained release of the neurotrophic factor neurotrophin-3 (NT-3) would support axonal plasticity in the spinal cord distal to the site of injury in rats. The corticospinal tract (CST) was cut unilaterally at the level of the medulla. This avoided excessive inflammation, secondary cell death, vascular disruption, and the release of inhibitory molecules in the lumbar spinal cord. A replication-defective adenoviral vector (Adv) carrying the NT-3 gene (Adv.EFalpha-NT3) was delivered to the spinal motoneurons by retrograde transport through the sciatic nerve. Retrograde transport of the adenoviral vectors avoided the inflammatory response that would be associated with direct injection into the spinal cord. Transduction of spinal motoneurons with Adv.EFalpha-NT3 resulted in a significant increase in the concentration of NT-3 in the L3-L6 region of the spinal cord for up to 3 weeks. In animals with a CST lesion, this local expression of NT-3 induced growth of axons from the intact CST across the midline to the denervated side. If the CST remained intact, overexpression of NT-3 did not lead to an increase in the number of axons crossing the midline. These data demonstrate that local, sustained expression of NT-3 will support axonal plasticity of intact CST axons after trauma-induced denervation.

Zhu, J. Y., Y. T. Huang, et al. (2003). "Expression of adenovirus-mediated neurotrophin-3 gene in Schwann cells of sciatic nerve in rats." Chin J Traumatol 6(2): 75-80.
 OBJECTIVE: To investigate the expression of neurotrophin-3 (NT-3) gene in Schwann cells of rat sciatic nerve introduced by an adenovirus vector in vivo. METHODS: A recombinant adenovirus vector for NT-3 (Ad-NT-3) was propagated in 293 packaging cells and titered with tissue culture infectious dose(50) (TCID(50)). Ad-NT-3 was injected directly into the rat sciatic nerve after transection and immediate repair. Immunohistochemical staining was employed to determine the expression of NT-3 in Schwann cells in rat sciatic nerve and the expressive intensity of the tissue slices of the sciatic nerve was measured with LEICA M550 image analysis system. RESULTS: On the 2nd day after injection of Ad-NT-3, positive stain in the Schwann cells was apparent in the vicinity of anastomosis. NT-3 expression increased significantly on the 7th day (P<0.01) and then decreased 14-28 days after injection (P<0.01). There was no significant difference of NT-3 expression between the 14th and 28th day groups (P<0.05). Compared with the 2nd day group, the 14th and 28th day groups still maintained a relatively high level of NT-3 (P<0.01). Intact and repaired nerves, which were injected with adenovirus encoding LacZ genes (Ad-LacZ) or physiological saline served as controls, showed no NT-3-positive Schwann cells. CONCLUSIONS: An adenovirus vector can be used to induce efficiently the expression of NT-3 gene in Schwann cells of rat peripheral nerves following nerve injury and repair, which suggests that neurotrophic factors can be introduced into Schwann cells with an adenovirus vector to promote peripheral nerve regeneration.

Zochodne, D. (2003). "Nerve regeneration and repair." J Peripher Nerv Syst 8(2): 59-60; author replies 61-2, 63-4.
 
Zupanc, G. K., S. C. Clint, et al. (2003). "Spatio-temporal distribution of microglia/macrophages during regeneration in the cerebellum of adult teleost fish, Apteronotus leptorhynchus: a quantitative analysis." Brain Behav Evol 62(1): 31-42.
 In contrast to mammals, adult teleost fish exhibit an enormous capacity to replace damaged neurons with newly generated ones after injuries in the central nervous system. In the present study, the role of microglia/macrophages, identified by tomato lectin binding, was examined in this process of neuronal regeneration in the corpus cerebelli of the teleost fish Apteronotus leptorhynchus. In the intact corpus cerebelli, or after short survival times following application of a mechanical lesion to this cerebellar subdivision, microglia/macrophages were virtually absent. Conversely, approximately 3 days after application of the lesion, the areal density of microglia/macrophages started to increase at and near the lesion site in the ipsilateral hemisphere, as well as in the contralateral hemisphere, and reached maximum levels at approximately 10 days post lesion. The density remained elevated until it reached background levels approximately one month after the injury. By comparing the time course of the appearance of microglia/macrophages with that of other regenerative events occurring within the first few weeks of wound healing in this model system, we hypothesize that one possible function of microglia/macrophages might be to remove debris of cells that have undergone apoptotic cell death at the lesion site.

Zupanc, G. K. and S. C. Clint (2003). "Potential role of radial glia in adult neurogenesis of teleost fish." Glia 43(1): 77-86.
 Persistence of radial glia within the adult central nervous system is a widespread phenomenon among fish. Based on a series of studies in the teleost species Apteronotus leptorhynchus, we propose that one function of this persistence is the involvement of radial glia in adult neurogenesis, i.e., the generation and further development of new neurons in the adult central nervous system. In particular, evidence has been obtained for the involvement of radial glia in the guidance of migrating young neurons in both the intact and the regenerating brain; for a possible role as precursor cells from which new neurons arise; and for its role as a source of trophic substances promoting the generation, differentiation, and/or survival of new neurons. These functions contribute not only to the potential of the intact brain to generate new neurons continuously, and of the injured brain to replace damaged cells by newly generated ones, but they also provide an essential part of the cellular substrate of behavioral plasticity.