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[en] PTEN inducible kinase-1 (PINK1) mutant induces mitochondrial dysfunction of cells, resulting in an inherited form of Parkinson's disease. However its exact role in the cardiomyocytes is unclear. The present study examined the function of PINK1 in hypoxia-reoxygenation (H/R) induced H9c2 cell damage and its potential mechanism. The H/R model in H9c2 cells was established by 6 h of hypoxia and 12 h of reoxygenation. The CCK8 and LDH assay indicated that the cell viability was obviously reduced after H/R. The expression of PINK1 was decreased in H/R-induced H9c2 cells compared with control group. The vector overexpressing PINK1 was constructed to transfect into H/R-induced H9c2 cells. Our results showed that cell viability was increased, cell apoptosis and caspase 3, cytochrome C (Cyto C) levels were decreased after LV-PINK1 transfection. Furthermore, PINK1 overexpression stabilized electron transport chain (ETC) activity, increased ATP production, mPTP opening and mitochondrial membrane potential (MMP), inhibited ROS-generating mitochondria, implying PINK1 alleviates H/R induced mitochondrial dysfunction in cardiomyocytes. In addition, the TRAP-1 siRNA was transfected into PINK1 treated H9c2 cells after H/R to detected the molecular mechanism of PINK1 protecting cardiomyocytes. The results indicated that silence of TRAP-1 reversed the effects of PINK1 in H/R-induced H9c2 cells. In conclusion, these results suggest that PINK1 overexpression alleviates H/R-induced cell damage of H9c2 cells by phosphorylation of TRAP-1, and that is a valid approach for protection from myocardial I/R injury. - Highlights: • Effects of H/R on cell viability and PINK1 expression in H9c2 cells. • Effects of PINK1 on cell viability in H9c2 cells with H/R model. • Effects of PINK1 on mitochondrial dysfunction in H9c2 cells with H/R model. • PINK1 ameliorates H/R-induced H9c2 cells injury by activating p-TRAP-1.
[en] The polyubiquitin genes Ubb and Ubc are upregulated under oxidative stress induced by arsenite [As(III)]. However, the role of ubiquitin (Ub) under As(III) exposure is not known in detail. In a previous study, we showed that the reduced viability observed in Ubc−/− mouse embryonic fibroblasts under As(III) exposure was not due to dysregulation of the Nrf2–Keap1 pathway, which prompted us to investigate another NFE2 family protein, nuclear factor erythroid 2-related factor 1 (Nrf1). In this study, we found that Ub deficiency due to Ubc knockdown in N2a cells reduced cell viability and proteasome activity under As(III) exposure. Furthermore, mRNA levels of the proteasome subunit Psma1 were also reduced. In addition, Ub deficiency led to the nuclear accumulation of the p65 isoform of Nrf1 under As(III) exposure. Interestingly, the overexpression of p65-Nrf1 recapitulated the phenotypes of Ub-deficient N2a cells under As(III) exposure. On the other hand, Nrf1 knockdown suppressed the death of Ub-deficient N2a cells upon exposure to As(III). Therefore, the levels of p65-Nrf1 may play an important role in the maintenance of cell viability under oxidative stress induced by As(III). - Highlights: • N2a cells exhibit reduced viability upon exposure to As(III) via Ubc knockdown. • As(III)-induced proteasomal regulation is impaired in Ub-deficient N2a cells. • Ub deficiency leads to the nuclear accumulation of p65-Nrf1 under As(III) exposure. • p65 expression recapitulates As(III)-induced phenotypes of Ub-deficient N2a cells. • Nrf1 knockdown suppressed As(III)-induced death of Ub-deficient N2a cells.
[en] Objective: To explore a method for isolating identifying and culturing the rat trachea-bronchia epithelial cells. Methods: The rat trachea-bronchia epithelial cells were isolated by digestion with pronase and brushing with cell brush, identified using confocul and cultured in entire F12 media with no serum. Results: With this method, cells in high purity and high viability could be obtained, and about 106 cells per rat. The cells grow well in entire F12 media with no serum. Conclusion: The method is useful for isolating rate trachea-bronchia epithelial cells and the entire F12 media with no serum is effective for culturing. (authors)
[en] Rural electrification (RE) has gained prominence over the past two decades as an effective means for improving living conditions. This growth has largely been driven by socio-economic and political imperatives to improve rural livelihood and by technological innovation. Based on a content analysis of 232 scholarly articles, the literature is categorized into four focal lenses: technology, institutional, viability and user-centric. We find that the first two dominate the RE debate. The viability lens has been used less frequently, whilst the user-centric lens began to engage scholars as late as 2007. We provide an overview of the technological, institutional and viability lenses, and elaborate upon the user-centric lens in greater detail. For energy policy and practice, we combine the four lenses to develop a business model framework that policy makers, practitioners and investors could use to assess RE projects or to design future rural electrification strategies. - Highlights: ► Review of two decades of rural electrification research. ► Content analysis of 232 scholarly articles. ► Literature is categorized into four focal lenses: technology, institutional, viability and user-centric. ► We develop a business model framework for rural electrification strategies.
[en] Labelling blood granulocytes presents two fields of interest: granulocyte kinetics and localization of deep-seated sites of infection. The authors have developed a new method of granulocyte separation and labelling. Its validity was tested in-vitro by a scanning electron microscope and by bactericidal and chemotactic assays. (orig.)
[en] Hsp90α's vital role in tumour survival and progression, together with its highly inducible expression profile in gliomas and its absence in normal tissue and cell lines validates it as a therapeutic target for glioma. Hsp90α was downregulated using the post-transcriptional RNAi strategy (sihsp90α) and a post-translational inhibitor, the benzoquinone antibiotic 17-AAG. Glioblastoma U87-MG and normal human astrocyte SVGp12 were treated with sihsp90α, 17-AAG and concurrent sihsp90α/17-AAG (combined treatment). Both Hsp90α gene silencing and the protein inhibitor approaches resulted in a dramatic reduction in cell viability. Results showed that sihsp90α, 17-AAG and a combination of sihsp90α/17-AAG, reduced cell viability by 27%, 75% and 88% (p < 0.001), respectively, after 72 h. hsp90α mRNA copy numbers were downregulated by 65%, 90% and 99% after 72 h treatment with sihsp90α, 17-AAG and sihsp90α/17-AAG, respectively. The relationship between Hsp90α protein expression and its client Akt kinase activity levels were monitored following treatment with sihsp90α, 17-AAG and sihsp90α/17-AAG. Akt kinase activity was downregulated as a direct consequence of Hsp90α inhibition. Both Hsp90α and Akt kinase levels were significantly downregulated after 72 h. Although, 17-AAG when used as a single agent reduces the Hsp90α protein and the Akt kinase levels, the efficacy demonstrated by combinatorial treatment was found to be far more effective. Combination treatment reduced the Hsp90α protein and Akt kinase levels to 4.3% and 43%, respectively, after 72 h. hsp90α mRNA expression detected in SVGp12 was negligible compared to U87-MG, also, the combination treatment did not compromise the normal cell viability. Taking into account the role of Hsp90α in tumour progression and the involvement of Akt kinase in cell signalling and the anti-apoptotic pathways in tumours, this double targets treatment infers a novel therapeutic strategy
[en] Stress echocardiography in low-dose dobutamine is part of the proposed techniques for the identification of viable myocardium. Unlike isotopic techniques that analyze the viability in terms of perfusion and membrane integrity, the echo dobutamine studying the functional aspect of sustain ability is - to - say the existence of a reservation contraction in segments asynergic. To establish the diagnostic value of echo dobutamine in search of sustain ability we conducted a prospective study to nearly 85 patients all presented with a recent myocardial infarction whose average age is 55 years and the sex ratio is 0.95. The examination was performed for all our patients on average twelfth day of myocardial infarction, improved segmental kinetics at least one grade in at least two adjacent segments relative to the ground state is considered as ultrasound test of the viability.
[en] The effect of irradiation of White Leghorn eggs with ultraviolet rays on their embryonal development, egg hatchability, viability of hatched chicks and their liveweight, is studied. Irradiation length was 1, 3, 5, 10, 15, 20, 40 and 60 min in two experiments and 2, 4, 16 and 256 min in one trial. It was established that egg irradiation with ultraviolet rays affected positively egg hatchability and viability of the chicks, the irradiation effect being strongest in the range of 2 to 40 min. No significant difference was established between liveweight of chicks obtained from irradiated and nonirradiated eggs. It was further found that the length of incubation period was shortened by 2 to 5 hrs with increase in irradiation length over 5 min. (author)