Localization and irregular distribution of Na,K-ATPase in myelin sheath from rat sciatic nerve

Sodium, potassium adenosine triphosphatase (Na,K-ATPase) is a membrane-bound enzyme that maintains the Na+ and K+ gradients used in the nervous system for generation and transmission of bioelectricity. Recently, its activity has also been demonstrated during nerve regeneration. The present study was undertaken to investigate the ultrastructural localization and distribution of Na,K-ATPase in peripheral nerve fibers. Small blocks of the sciatic nerves of male Wistar rats weighing 250-300g were excised, divided into two groups, and incubated with and without substrate, the para-nitrophenyl phosphate (pNPP). The material was processed for transmission electron microscopy, and the ultra-thin sections were examined in a Philips CNI 100 (TM) electron microscope. The deposits of reaction product were localized mainly on the axolemma, on axoplasmic profiles, and irregularly dispersed on the myelin sheath, but not in the unmyelinated axons. In the axonal membrane, the precipitates were regularly distributed on the cytoplasmic side. These results together with published data warrant further studies for the diagnosis and treatment of neuropathies with compromised Na,K-ATPase activity. (c) 2007 Elsevier Ltd. All rights reserved.

GENETIC INSTABILITY IN NERVE SHEATH CELL TUMORS

After in vitro culture, we analyzed cytogenetically four acoustic nerve neurinomas, one intraspinal neurinoma and one neurofibroma obtained from unrelated patients. Monosomy of chromosomes 22 and 16 was an abnormality common to all cases, followed in frequency by loss of chromosomes 18 (three cases) and chromosomes 8, 17 and 19 (two cases). Trisomy of chromosome 20 was also detected in two cases. Structural rearrangements were detected at low frequencies, with del(10)(p12) being present in two cases. In addition, we observed cell subpopulations showing a certain degree of genetic instability, reflected by the presence of polyploid cells with inconsistent abnormalities, endoreduplications and telomeric associations resulting in dicentric chromosomes. It is probable that these cytogenetic abnormalities represent some kind of evolutionary advantage for the in vitro progression of nerve sheath tumors.

The use of nonparametric contrasts in one-way layouts and random block designs

Nonparametric simple-contrast estimates for one-way layouts based on Hodges-Lehmann estimators for two samples and confidence intervals for all contrasts involving only two treatments are found in the literature.Tests for such contrasts are performed from the distribution of the maximum of the rank sum between two treatments. For random block designs, simple contrast estimates based on Hodges-Lehmann estimators for one sample are presented. However, discussions concerning the significance levels of more complex contrast tests in nonparametric statistics are not well outlined.This work aims at presenting a methodology to obtain p-values for any contrast types based on the construction of the permutations required by each design model using a C-language program for each design type. For small samples, all possible treatment configurations are performed in order to obtain the desired p-value. For large samples, a fixed number of random configurations are used. The program prompts the input of contrast coefficients, but does not assume the existence or orthogonality among them.In orthogonal contrasts, the decomposition of the value of the suitable statistic for each case is performed and it is observed that the same procedure used in the parametric analysis of variance can be applied in the nonparametric case, that is, each of the orthogonal contrasts has a chi(2) distribution with one degree of freedom. Also, the similarities between the p-values obtained for nonparametric contrasts and those obtained through approximations suggested in the literature are discussed.

Complete staining of nerve fiber and myoneural junctions with acetylthiocholine and silver

We describe a combined stain for simultaneous demonstration of the preterminal axons and cholinesterase activity at myoneural junctions of mammalian muscles. This technique employs acetylthiocholine iodide as the substrate for cholinesterase activity and silver nitrate impregnation of preterminal axons. The procedure is rapid, simple and uses fresh muscles. Intramuscular nerves, preterminal axons and myoneural junctions are stained simultaneously brown or black with minimal background staining of connective tissue and muscle fibers.