Neuroprotective Effects of Withania somnifera Dunal.: A Possible Mechanism

Present study was carried out to understand the possible mechanism of neuroprotective action of the root extract of Withania somnifera Dunal (WS). The study is focused on WS mediated inhibition of nitric oxide production, which is known to mediate neurodegeneration during stress. Adult mice (28 ± 5 g) were exposed to restraint stress for 30 days. Activity of NADPH diaphorase (NADPH-d) and factors (Acetylcholine, serotonin and corticosterone), which regulates NADPH-d activity were studied. Treatment with WS extract for 30 days during stress, significantly reversed the stress induced NADPH-d activation. Observations suggest that inhibition of NADPH-d by WS is not a direct effect of extract on NADPH-d, instead it inhibits via suppressing corticosterone release and activating cholineacetyltransferase, which in turn increase serotonin level in hippocampus to inhibit NADPH-d. Together, the main mechanism underlying the neuroprotective effects of WS can be attributed to its role in the down regulation of nNOS and neurochemical alterations of specific neurotransmitter systems. These observations thus suggest that WS root extract could be developed as a potential preventive or therapeutic drug for stress induced neurological disorders.     

Acute and transient effects of stress on immunoreactive somatostatin cell bodies and fibers in the preoptic-anterior hypothalamus and median eminence of female rats

Stress in rats causes acute release of hypothalamic somatostatin (SS) in median eminence (ME) that induces a marked and prolonged suppression of growth hormone (GH) secretion. This was evidenced by immunocytochemistry (ICC) and radioimmunoassay (RIA) in the present study. Adult female rats were decapitated under nonstress or for 30, 60, 120 and 180 min after 15 min leg restraint stress. The rabbit anti-SS was used to detect SS-14 and SS-28 containing cell bodies with ICC in preoptic-anterior hypothalamus (PO-AH). At 30, 60, 120 min after stress, there was marked decrease in the number and size of subsets of SS cell bodies. RIA demonstrated striking increase in SS in ME and significant decrease in GH of the portal blood. The most reproducible changes in cell bodies involved subsets of PeV neurons. Interestingly, these changes were largely reversed by 180 min. The results of the study demonstrate that stress cause acute changes in PO-AH, SS system and it appears that stress affects both SS synthesis and the secretion.     

Two-dimensional electrophoretic analysis of major histone species and their variants from somatic and germ-line tissue

A two-dimensional electrophoretic procedure has been developed and applied to the analysis of histones from the mouse thymus, liver, and seminiferous epithelium. The technique uses acetic acid-urea polyacrylamide gel electrophoresis in the first dimension to provide a primary separation of major histone species. Separation of additional histone species and variants is achieved in the second dimension by adding 0.4% of the nonionic detergent Lubrol-WX to the polyacrylamide gel. The procedure is relatively simple and highly reproducible and enables the simultaneous resolution of 9 to 16 protein spots corresponding to the major histone species and their variants.     

Evidence for lysogeny and viral resistance in the cyanobacteriumPhormidium uncinatum

Escherichia coli B/5 12-h cultures were exposed to filter-sterilized acid mine water (AMW), fixed in situ, and examined for morphological changes by transmission electron microscopy, scanning electron microscopy, and x-ray spectrometry. Thin sections showed that layers of the Gram-negative envelope were altered and often lacking. Additionally, polar regions of the cell were frequently devoid of cytoplasm. AMW-exposed cells were distorted and had an amorphous substance associated with them. Spectra obtained by x-ray spectrometry suggested that this amorphous substance was cytoplasm rather than a mineral precipitate from AMW. Morphometric analyses of control and AMW-exposed populations showed significant differences in mean volume, length, and width of cells stressed in AMW; this indicates that smaller cells were selectively destroyed by the action of AMW. We concluded that loss of cytoplasm and cell lysis were the consequence of AMW damage to the bacterial envelope.     

Exploitation of the broad specificity of the membrane-bound isoenzyme of lactate dehydrogenase for direct selection of null mutants in Neisseria gonorrhoeae

Lactic acid is readily utilized as a carbon and energy source by Neisseria gonorrhoeae. The oxidation of lactate is coupled to electron transport via a membrane-bound lactate dehydrogenase (iLDH) which is independent of pyridine nucleotide. The broad substrate specificity of iLDH endows N. gonorrhoeae with the novel ability to convert phenyllactate to L-phenylalanine via phenylpyruvate. N. gonorrhoeae ATCC 27628 typifies a class of clinical isolate whose growth is inhibited by phenylpyruvate (or L-phenylalanine). Exploiting resistance to growth inhibition by phenyllactate as a strategy of positive selection, mutant derivatives of strain ATCC 27628 lacking iLDH activity were readily obtained. These mutants are incapable of oxidizing phenyllactate, and lack the parent-strain ability to reduce c-type cytochromes in the presence of lactate, phenyllactate or 4-hydroxyphenyllactate. They retain, however, a cytoplasmic NAD(+)-linked lactate dehydrogenase (nLDH). Since the mutants retained the ability to grow on lactate as a sole source of carbon, nLDH presumably can function in an opposite-to-normal physiological direction in the absence of iLDH. This would explain the failure to isolate iLDH-deficient mutants by selection for inability to grow on lactate.     

Characterization of the defect in the Escherichia coli mutT1 mutator gene.

With a probe constructed from the wild-type gene, a DNA fragment containing the entire mutT1 mutator gene was isolated and cloned into pUC18. Nucleotide sequence analysis revealed that the mutator defect was most likely due to an IS1 insertion into the wild-type gene.