Nerve Specimens (nerve + specimen)

Distribution by Scientific Domains


Selected Abstracts


HCV-RNA In Sural Nerve From Hcv Infected Patients With Peripheral Neuropathy

JOURNAL OF THE PERIPHERAL NERVOUS SYSTEM, Issue 1 2001
L De Martino
Objective: Evaluation of hepatitis C virus (HCV) by reverse transcription-polymerase chain reaction (RT-PCR) in peripheral nerve tissues from HCV infected patients with peripheral neuropathy. METHODS: RT-PCR was performed on homogenates of nerve biopsies from 17 consecutive HCV-positive patients with peripheral neuropathy, with or without mixed cryoglobulinemia, hospitalised from 1996 to 2000. Sural nerve specimens were frozen in iso-pentane pre-cooled in liquid nitrogen and stored at ,80°C until use. RNA was extracted from ten 7-,m thick cryostatic sections or from a nerve trunk specimen of about 3 mm length, collected from each biopsy. Three different protocols of RNA extraction were tested (1,3). Complementary DNAs (cDNAs) were obtained without or with RNasin (Promega, Madison, WI) addition in the reaction mixture to inhibit residual RNase activity. Two sets of commercially available PCR primers for the outer and the nested reaction were used. PCR products were analysed by agarose gel electrophoresis and ethidium bromide staining. Serum samples and liver specimens from proven HCV positive patients served as positive controls, whereas sera from healthy subjects were negative controls. RESULTS: Sufficient amount of RNA could be obtained either by cryostatic sections or by in toto nerve specimens. Extraction by Trizol (Gibco-BRL) allowed the best concentration and purity of RNA as assessed by biophotometry. The presence of RNasin didn't improve the cDNA synthesis. The resulting amplification product of the nested PCR was 187 bp long. We have always observed this product in our positive controls and never in the negative. Six samples from patients either with or without cryoglobulinemia resulted positive; 7 were negative. Four samples gave variable results. CONCLUSIONS: While 40% of the nerves in our series were undoubtedly HCV positive, the cause(s) of negative and variable results in the remaining samples is likely more complex than variations in the detection protocols and deserve further investigations. REFERENCES: 1) Chomczynski P, Sacchi N (1987). Anal Biochem 162:156. 2) Marquardt O et al. (1996). Med Microbiol Lett 5:55. 3) Chomczynski P (1993). Bio/Techniques 15:532. [source]


MACROPHAGE INFILTRATION AND INDUCTION OF P75 NTR AND IL-1B IN THE NERVE OF DIABETIC RATS

JOURNAL OF THE PERIPHERAL NERVOUS SYSTEM, Issue 1 2000
G. Conti
Recently, inflammation has been involved in the pathogenesis of diabetic neuropathy, and activated macrophages have been found in the peripheral nervous system of diabetic rats, with a possible role in chemotaxis and regeneration. In this study, we obtained sciatic nerve specimens from diabetic rats at different time points following STZ administration. Macrophages infiltration, IL-1b and p75NTR induction were analyzed by immunocytochemistry on frozen sections and on teased nerve fibers. Apoptosis was detected on teased nerve fibers by TUNEL and DAPI staining. Cell phenotype was characterized by double-staining with antibodies specific for Schwann cells and macrophages. The nerves obtained from STZ-diabetic rats showed macrophages infiltration by day 14 following STZ administration, with complete clearance by day 35. Fifteen percent of these cells were TUNEL positive. IL-1B induction was concomitant with macrophages infiltration and not detectable by day 35. p75NTR expression began by day 21, peaking by day 35, and dropping to barely detectable levels by day 105. These findings seem to indicate that the concomitance of these processes may be crucial in the regulation of nerve damage and in promoting an attempt of regeneration at the early stages of STZ diabetic neuropathy. [source]


A simple protocol for paraffin-embedded myelin sheath staining with osmium tetroxide for light microscope observation

MICROSCOPY RESEARCH AND TECHNIQUE, Issue 7 2008
Federica Di Scipio
Abstract Experimental investigation of peripheral nerve fiber regeneration is attracting more and more attention among both basic and clinical researchers. Assessment of myelinated nerve fiber morphology is a pillar of peripheral nerve regeneration research. The gold standard for light microscopic imaging of myelinated nerve fibers is toluidine blue staining of resin-embedded semithin sections. However, many researchers are unaware that the dark staining of myelin sheaths typically produced by this procedure is due to osmium tetroxide postfixation and not due to toluidine blue. In this article, we describe a simple pre-embedding protocol for staining myelin sheaths in paraffin-embedded nerve specimens using osmium tetroxide. The method involves immersing the specimen in 2% osmium tetroxide for 2 h after paraformaldeyde fixation, followed by routine dehydration and paraffin embedding. Sections can then be observed directly under the microscope or counterstained using routine histological methods. Particularly good results were obtained with Masson's trichrome counterstain, which permits the imaging of connective structures in nerves that are not detectable in toluidine blue-stained resin sections. Finally, we describe a simple protocol for osmium etching of sections, which makes further immunohistochemical analysis possible on the same specimens. Taken together, our results suggest that the protocol described in this article is a valid alternative to the conventional resin embedding-based protocol: it is much cheaper, can be adopted by any histological laboratory, and allows immunohistochemical analysis to be conducted. Microsc. Res. Tech., 2008. © 2008 Wiley-Liss, Inc. [source]


Alcoholic neuropathy is clinicopathologically distinct from thiamine-deficiency neuropathy

ANNALS OF NEUROLOGY, Issue 1 2003
Haruki Koike MD
Characteristics of alcoholic neuropathy have been obscured by difficulty in isolating them from features of thiamine-deficiency neuropathy. We assessed 64 patients with alcoholic neuropathy including subgroups without (ALN) and with (ALN-TD) coexisting thiamine deficiency. Thirty-two patients with nonalcoholic thiamine-deficiency neuropathy (TDN) also were investigated for comparison. In ALN, clinical symptoms were sensory-dominant and slowly progressive, predominantly impairing superficial sensation (especially nociception) with pain or painful burning sensation. In TDN, most cases manifested a motor-dominant and acutely progressive pattern, with impairment of both superficial and deep sensation. Small-fiber-predominant axonal loss in sural nerve specimens was characteristic of ALN, especially with a short history of neuropathy; long history was associated with regenerating small fibers. Large-fiber-predominant axonal loss predominated in TDN. Subperineurial edema was more prominent in TDN, whereas segmental de/remyelination resulting from widening of consecutive nodes of Ranvier was more frequent in ALN. Myelin irregularity was greater in ALN. ALN-TD showed a variable mixture of these features in ALN and TDN. We concluded that pure-form of alcoholic neuropathy (ALN) was distinct from pure-form of thiamine-deficiency neuropathy (TDN), supporting the view that alcoholic neuropathy can be caused by direct toxic effect of ethanol or its metabolites. However, features of alcoholic neuropathy is influenced by concomitant thiamine-deficiency state, having so far caused the obscure clinicopathological entity of alcoholic neuropathy. Ann Neurol 2003 [source]