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Previous studies using the Hep G2-based VA cells showed that ethanol metabolism resulted in both cytotoxicity and impaired DNA synthesis, causing reduced accumulation of cells in culture. To further characterize the ethanol oxidation-mediated impairment of DNA synthesis we analyzed the cell-cycle progression of VA cells. These studies showed approximately a 6-fold increase in the percentage of cells in the G2/M phase of the cell cycle after 4 days of ethanol exposure. The G2/M transition requires activity of the cyclin-dependent kinase, Cdc2. Cdc2 is positively regulated by association with cyclin B1, and negatively regulated by phosphorylation of amino acids Thr14 and Tyr15. Immunoblot analysis revealed that ethanol metabolism had little affect on total Cdc2 content in these cells, but resulted in the accumulation of up to 20 times the amount of cyclin B1, indicating that cyclin B1 was available for formation of Cdc2/cyclin B1 complexes. Co-immunoprecipitation revealed that 6 times more Cdc2/cyclin B1 complexes were present in the ethanol-treated cells compared with the controls. Investigation of the phosphorylation state of Cdc2 revealed that ethanol oxidation increased the amount of the phosphorylated inactive form of Cdc2 by approximately 3-fold. Thus, the impairment in cell-cycle progression could not be explained by a lack of cyclin B1, or the ability of Cdc2 and cyclin B1 to associate, but instead resulted, at least in part, from impaired Cdc2 activity. In conclusion, ethanol oxidation by VA cells results in a G2/M cell-cycle arrest, mediated by accumulation of the phosphorylated inactive form of Cdc2. 相似文献
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Case history, light and electron microscopic findings of a case of a lymphocytic hypophysitis in coincidence with a suprasellar germinoma in a 12-year-old girl are reported. The girl presented with a long time case history of diabetes insipidus and subsequent panhypopituitarism. Two years after the diagnosis of diabetes insipidus magnetic resonance imaging (MRI) showed a tumorous enlargement of the sellar content and pituitary stalk. A transnasal exploration was initially performed and revealed a lymphocytic hypophysitis. Light microscopy showed a dense infiltration of mature lymphocytes and plasma cells in the interstitium of the anterior pituitary gland. The stalk area could not be exposed to exclude a germinoma. One year later the lesion relapsed despite dexamethason therapy and a second operation by another neurosurgeon had to be performed. Light microscopy showed lymphocytic infiltrates, fibrosis and necrosis. The diagnosis was a lymphocytic hypophysitis again. Though transcranially exposed only pituitary tissue was removed. No infundibular mass became visible at surgery as shown by MRI. The girl developed five months later multiple cerebral lesions, which revealed to be a germinoma. Lymphocytic hypophysitis in children is very rare and a coincedence with a germinoma has not been described from histopathological aspect until now. The origin of the pituitary infiltration is discussed. 相似文献
96.
Mittmann C Pinkepank G Stamatelopoulou S Wieland T Nürnberg B Hirt S Eschenhagen T 《Journal of molecular and cellular cardiology》2003,35(10):1241-1249
Muscarinic acetylcholine receptors (mAChRs) mediate their main cardiac effects via pertussis toxin-sensitive G-proteins. Physiological effects differ considerably between atrium and ventricle, and it is unknown to which extent these differences derive from selective receptor-G-protein coupling or further downstream events. We have characterized specific coupling between mAChRs and Gi/Go-protein isoforms in atrial and ventricular myocardium by agonist-dependent photoaffinity labeling with [(32)P]azidoanilido GTP (aaGTP) and immunoprecipitation in sarcolemmal membranes from terminally failing human hearts. The total amount of mAChRs, as determined by specific binding of [(3)H]QNB, was significantly higher in right-atrial (RA +/- SEM, 959 +/- 68 fmol/mg, n = 4) than in left-ventricular membranes (LV, 582 +/- 53 fmol/mg, n = 6). Standardized immunoblots revealed that Gialpha-2 was the predominant subtype in both regions. A 40-kDa splice variant of Goalpha (Goalpha-1 and/or Goalpha-3) was almost exclusively detectable in RA. Levels of Gialpha-3 and a 39-kDa splice variant of Goalpha (Goalpha-2) were also higher in RA. Basal aaGTP binding was higher in RA than in LV for all Gialpha/Goalpha subtypes. The carbachol (10 micromol/l)-induced increase in aaGTP binding was significantly higher in RA than in LV for Goalpha-1/3 (336 +/- 95% of LV, n = 4) and for Gialpha-3 (211 +/- 83%), lower for Gialpha-2 (42 +/- 5%), and was similar in both regions for Goalpha-2 (130 +/- 62%). The differential coupling of mAChRs in human RA and LV suggests that the initiation of different physiological responses to mAChR stimulation starts with signal sorting at the receptor-G-protein level. 相似文献
97.
New insights into the effects of biomaterial chemistry and topography on the morphology of kidney epithelial cells 下载免费PDF全文
Frits Hulshof Carolien Schophuizen Milos Mihajlovic Clemens van Blitterswijk Rosalinde Masereeuw Jan de Boer Dimitrios Stamatialis 《Journal of tissue engineering and regenerative medicine》2018,12(2):e817-e827
Increasing incidence of renal pathology in the western world calls for innovative research for the development of cell‐based therapies such as a bioartificial kidney (BAK) device. To fulfil the multitude of kidney functions, the core component of the BAK is a living membrane consisting of a tight kidney cell monolayer with preserved functional organic ion transporters cultured on a polymeric membrane surface. This membrane, on one side, is in contact with blood and therefore should have excellent blood compatibility, whereas the other side should facilitate functional monolayer formation. This work investigated the effect of membrane chemistry and surface topography on kidney epithelial cells to improve the formation of a functional monolayer. To achieve this, microtopographies were fabricated with high resolution and reproducibility on polystyrene films and on polyethersulfone‐polyvinyl pyrrolidone (PES‐PVP) porous membranes. A conditionally immortalized proximal tubule epithelial cell line (ciPTEC) was cultured on both, and subsequently, the cell morphology and monolayer formation were assessed. Our results showed that L‐dopamine coating of the PES‐PVP was sufficient to support ciPTEC monolayer formation. The polystyrene topographies with large features were able to align the cells in various patterns without significantly disruption of monolayer formation; however, the PES‐PVP topographies with large features disrupted the monolayer. In contrast, the PES‐PVP membranes with small features and with large spacing supported well the ciPTEC monolayer formation. In addition, the topographical PES‐PVP membranes were compatible as a substrate membrane to measure organic cation transporter activity in Transwell® systems. Copyright © 2016 John Wiley & Sons, Ltd. 相似文献
98.
Xinlei Wu Clemens von Birgelen Zehang Li Su Zhang Jiayue Huang Fuyou Liang Yingguang Li William Wijns Shengxian Tu 《The international journal of cardiovascular imaging》2018,34(6):849-861
Cyclic biomechanical stress at the lumen-intima interface plays a crucial role in the rupture of coronary plaque. We performed a comprehensive assessment of a novel angiography-based method for four-dimensional (4D) dynamic assessment of superficial wall stress (SWS) and deformation with a total of 32 analyses in virtual stenosis models with equal lumen dimensions and 16 analyses in human coronary arteries in vivo. The in silico model analyses demonstrated that the SWS, derived by the proposed global displacement method without knowledge of plaque components or blood pressure, was comparable with the result calculated by traditional finite element method. Cardiac contraction-induced vessel deformation increased SWS. Softer plaque and positive arterial remodeling, associated with a greater plaque burden, showed more variation in mean lumen diameter within the cardiac cycle and resulted in higher SWS. In vivo patient analyses confirmed the accuracy of computed superficial wall deformation. The centerlines predicted by our method at random selected time instant matched well with the actual one in angiograms by Procrustes analysis (scaling: 0.995?±?0.018; dissimilarity: 0.007?±?0.014). Over 50% of the maximum SWS occurred at proximal plaque shoulders. This novel 4D approach could be successfully to predict superficial wall deformation of coronary artery in vivo. The dynamic SWS might be more realistic to evaluate the risk of plaque rupture. 相似文献
99.
Roger Clemens Adam Drewnowski Mario G Ferruzzi Cheryl D Toner Diane Welland 《Advances in nutrition (Bethesda, Md.)》2015,6(2):236S-243S
Total fruit intake in the United States is ~1 cup equivalent per day, or one-half of the 2010 Dietary Guidelines for Americans recommendation for adults. Two-thirds of the fruit consumed is whole fruit and one-third is 100% juice. The nutritional value of whole fruit, with the exception of fiber and vitamin C, may be retained with appropriate juice production methods and storage conditions. One-hundred percent fruit juice consumption is associated with a number of health benefits, such as improved cardiovascular health and decreased obesity, although some of these and other potential benefits are controversial. Comprehensive analyses of the evidence by the Academy of Nutrition and Dietetics in 2014, the US Dietary Guidelines Advisory Committee in 2010, and the Australian Dietary Guidelines of 2013 concluded that 100% fruit juice is not related to adiposity in children when consumed in appropriate amounts for age and energy needs. However, some reports suggest the consumption of fruit juice contributes to unhealthful outcomes, particularly among children. A dietary modeling study on the best ways to meet the fruit intake shortfall showed that a combination of whole fruit and 100% juice improved dietary density of potassium and vitamin C without significantly increasing total calories. Notably, 100% juice intake was capped at amounts consistent with the 2001 American Pediatric Association guidance. The preponderance of evidence supports the position that 100% fruit juice delivers essential nutrients and phytonutrients, provides year-round access to a variety of fruits, and is a cost-effective way to help people meet fruit recommendations. 相似文献