Circadian rhythm in acetylcholinesterase activity during aging of the central nervous system

The circadian rhythm of acetylcholinesterase (AChE) activity was investigated in the cerebrum, cerebellum, medulla and optic lobes of rats aged 1 day and 3, 13, 44 and 87 weeks. The rhythm was found to be age-dependent. Animals aged 1 day exhibited a bimodal rhythm in all the four regions of the brain studied. At 3 and 13 weeks the activity was unimodal. The peak occurred during the light-on phase at 3 weeks and during the light-off phase at 13 weeks. At 87 weeks the rhythms in the medulla and cerebrum were similar to those of 44 week animals. By contrast the cerebellum had a bimodal rhythm with peaks at intervals of 12 h. In the optic lobes there was a shift from a bimodal pattern at 44 weeks to a unimodal one at 87 weeks. The times of onset of the light-on and light-off peaks in different regions of the brain differed with age.     

Roles of Pdk1p, a fission yeast protein related to phosphoinositide-dependent protein kinase, in the regulation of mitosis and cytokinesis

Proteins related to the phosphoinositide-dependent protein kinase family have been identified in the majority of eukaryotes. Although much is known about upstream mechanisms that regulate the PDK1-family of kinases in metazoans, how these kinases regulate cell growth and division remains unclear. Here, we characterize a fission yeast protein related to members of this family, which we have termed Pdk1p. Pdk1p localizes to the spindle pole body and the actomyosin ring in early mitotic cells. Cells deleted for pdk1 display multiple defects in mitosis and cytokinesis, all of which are exacerbated when the function of fission yeast polo kinase, Plo1p, is partially compromised. We conclude that Pdk1p functions in concert with Plo1p to regulate multiple processes such as the establishment of a bipolar mitotic spindle, transition to anaphase, placement of the actomyosin ring and proper execution of cytokinesis. We also present evidence that the effects of Pdk1p on cytokinesis are likely mediated via the fission yeast anillin-related protein, Mid1p, and the septation initiation network.     

The Functional Potential of Microbial Communities in Hydraulic Fracturing Source Water and Produced Water from Natural Gas Extraction Characterized by Metagenomic Sequencing

Microbial activity in produced water from hydraulic fracturing operations can lead to undesired environmental impacts and increase gas production costs. However, the metabolic profile of these microbial communities is not well understood. Here, for the first time, we present results from a shotgun metagenome of microbial communities in both hydraulic fracturing source water and wastewater produced by hydraulic fracturing. Taxonomic analyses showed an increase in anaerobic/facultative anaerobic classes related to Clostridia, Gammaproteobacteria, Bacteroidia and Epsilonproteobacteria in produced water as compared to predominantly aerobic Alphaproteobacteria in the fracturing source water. The metabolic profile revealed a relative increase in genes responsible for carbohydrate metabolism, respiration, sporulation and dormancy, iron acquisition and metabolism, stress response and sulfur metabolism in the produced water samples. These results suggest that microbial communities in produced water have an increased genetic ability to handle stress, which has significant implications for produced water management, such as disinfection.     

Effect of phenol on protein and amino acid content of Xanthomonas oryzae pv. oryzae

Leaf blight disease of rice (Oryza sativa) is caused by the bacterium Xanthomonas oryzae pv. oryzae. Phenol (1 to 4 mM) induced changes in protein profiles of X. o. pv. oryzae and a stress protein with a molecular mass of 69,000 appeared. HPLC analysis indicated occurrence of amino acids such as asparagine, alanine, methionine and cystine in phenol treated cells. Proton NMR analysis also revealed variation on the presence of amino acids in the cells treated with phenol.     

Pre-exposure of calli to ozone promotes tolerance of regenerated Lycopersicon esculentum cv. PKM1 plantlets against acute ozone stress

Studies were performed to evaluate the effects of pre-exposure of calli to ozone in promoting tolerance of the regenerated Lycopersicon esculentum cv. PKM1 (tomato) plantlets against acute ozone stress (AOS). Calli induced from tomato leaf explants were subjected to pre-treatment with ozone: T(1)=100ppb, T(2)=200ppb and T(3)=300ppb. For the control (C) calli, charcoal-filtered air was supplied to test differential sensitivity of regenerated plantlets to acute ozone stress. All treated calli were subsequently transferred to shooting, rooting medium and acclimatized. The plantlets regenerated from the respective ozone (T(1), T(2), T(3))-treated calli are referred to here as T(1), T(2), T(3) plantlets and the plantlets regenerated from control calli are referred to as control plantlets. The frequencies of regeneration of tomato plantlets from the calli were T(1)=86%, T(2)=82% and T(3)=67%, and 92% regeneration was obtained from control calli. In order to evaluate the ozone tolerance, all the regenerated plantlets were exposed to the acute ozone exposure (AOE). After AOE, the T(2) plantlets endured remarkably well by experiencing reduced ozone stress, which was evident from the lower level of hydrogen peroxide and oxidative stress-related enzymes such as ascorbate peroxidase (EC 1.11.1.11) and superoxide dismutase (EC 1.15.1.1) activities relative to T(3), T(1) and C plantlets. All T(2) plantlets showed enhanced tolerance against AOE by upholding enhanced soluble phenol content, a higher level of foliar and apoplastic ascorbic acid, elevated dehydroascorbate reductase (EC 1.8.5.1) and glutathione content. The present study reveals that the calli pre-exposed to T(2) ozone treatment resulted in an increase in the level of antioxidants and provided the plants greater protection against acute ozone stress.     

Corneal antifibrotic switch identified in genetic and pharmacological deficiency of vimentin

The type III intermediate filaments (IFs) are essential cytoskeletal elements of mechanosignal transduction and serve critical roles in tissue repair. Mice genetically deficient for the IF protein vimentin (Vim(-/-)) have impaired wound healing from deficits in myofibroblast development. We report a surprising finding made in Vim(-/-) mice that corneas are protected from fibrosis and instead promote regenerative healing after traumatic alkali injury. This reparative phenotype in Vim(-/-) corneas is strikingly recapitulated by the pharmacological agent withaferin A (WFA), a small molecule that binds to vimentin and down-regulates its injury-induced expression. Attenuation of corneal fibrosis by WFA is mediated by down-regulation of ubiquitin-conjugating E3 ligase Skp2 and up-regulation of cyclin-dependent kinase inhibitors p27(Kip1) and p21(Cip1). In cell culture models, WFA exerts G(2)/M cell cycle arrest in a p27(Kip1)- and Skp2-dependent manner. Finally, by developing a highly sensitive imaging method to measure corneal opacity, we identify a novel role for desmin overexpression in corneal haze. We demonstrate that desmin down-regulation by WFA via targeting the conserved WFA-ligand binding site shared among type III IFs promotes further improvement of corneal transparency without affecting cyclin-dependent kinase inhibitor levels in Vim(-/-) mice. This dissociates a direct role for desmin in corneal cell proliferation. Taken together, our findings illuminate a previously unappreciated pathogenic role for type III IF overexpression in corneal fibrotic conditions and also validate WFA as a powerful drug lead toward anti-fibrosis therapeutic development.     

In vitro morphogenic response in cotyledon explants of Semecarpus anacardium L.

Three different morphogenic responses??caulogenesis, direct somatic embryogenesis, and callusing??were noted in cotyledon explants of Semecarpus anacardium L. cultured in woody plant medium (WPM) containing thidiazuron (TDZ). Thidiazuron, at all concentrations tested, induced organogenic as well as embryogenic responses. The organogenic buds differentiated to shoots and the embryogenic mass (EM) gave rise to globular embryos which differentiated up to cotyledon-stage embryos on repeated culture in growth regulator (GR)-free WPM medium containing 0.2% activated charcoal after the removal of TDZ. The organogenic and embryogenic responses were optimal in 9.08???M TDZ after the removal of TDZ. Elongated shoots rooted in half-strength liquid WPM medium with 2.46???M indole butyric acid. Plants were successfully acclimatized and transferred to soil. Histological studies confirmed the direct origin of the organogenic buds from the cotyledon explants. The EMs produced somatic embryos on repeated culture in charcoal incorporated GR-free medium. Morphogenic callus formation from the cotyledon explants was also noted. This callus on repeated culture in WPM medium with charcoal differentiated into somatic embryos. Repetitive somatic embryogenesis was evident from direct and indirectly formed primary embryos. The somatic embryos did not convert into plantlets, though sporadic germination of embryos was observed through the emergence of roots.