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81.
Grégory Lacraz Florence Figeac Jamileh Movassat Nadim Kassis Josiane Coulaud Anne Galinier Corinne Leloup Danielle Bailbé Fran?oise Homo-Delarche Bernard Portha 《PloS one》2009,4(8)
Background
Oxidative stress (OS), through excessive and/or chronic reactive oxygen species (ROS), is a mediator of diabetes-related damages in various tissues including pancreatic β-cells. Here, we have evaluated islet OS status and β-cell response to ROS using the GK/Par rat as a model of type 2 diabetes.Methodology/Principal Findings
Localization of OS markers was performed on whole pancreases. Using islets isolated from 7-day-old or 2.5-month-old male GK/Par and Wistar control rats, 1) gene expression was analyzed by qRT-PCR; 2) insulin secretion rate was measured; 3) ROS accumulation and mitochondrial polarization were assessed by fluorescence methods; 4) antioxidant contents were quantified by HPLC. After diabetes onset, OS markers targeted mostly peri-islet vascular and inflammatory areas, and not islet cells. GK/Par islets revealed in fact protected against OS, because they maintained basal ROS accumulation similar or even lower than Wistar islets. Remarkably, GK/Par insulin secretion also exhibited strong resistance to the toxic effect of exogenous H2O2 or endogenous ROS exposure. Such adaptation was associated to both high glutathione content and overexpression (mRNA and/or protein levels) of a large set of genes encoding antioxidant proteins as well as UCP2. Finally, we showed that such a phenotype was not innate but spontaneously acquired after diabetes onset, as the result of an adaptive response to the diabetic environment.Conclusions
The GK/Par model illustrates the effectiveness of adaptive response to OS by β-cells to achieve self-tolerance. It remains to be determined to what extend such islet antioxidant defenses upregulation might contribute to GK/Par β-cell secretory dysfunction. 相似文献82.
Significant differentiation in the apolipoprotein(a)/lipoprotein(a) trait between chimpanzees from Western and Central Africa 下载免费PDF全文
83.
84.
85.
Anne Chevrel Agnes Mesneau Dyana Sanchez Louisa Celma Sophie Quevillon-Cheruel Andrea Cavagnino Sylvie Nessler Ines Li de la Sierra-Gallay Herman van Tilbeurgh Philippe Minard Marie Valerio-Lepiniec Agathe Urvoas 《Journal of structural biology》2018,201(2):88-99
We have previously described a highly diverse library of artificial repeat proteins based on thermostable HEAT-like repeats, named αRep. αReps binding specifically to proteins difficult to crystallize have been selected and in several examples, they made possible the crystallization of these proteins. To further simplify the production and crystallization experiments we have explored the production of chimeric proteins corresponding to covalent association between the targets and their specific binders strengthened by a linker. Although chimeric proteins with expression partners are classically used to enhance expression, these fusions cannot usually be used for crystallization. With specific expression partners like a cognate αRep this is no longer true, and chimeric proteins can be expressed purified and crystallized. αRep selection by phage display suppose that at least a small amount of the target protein should be produced to be used as a bait for selection and this might, in some cases, be difficult. We have therefore transferred the αRep library in a new construction adapted to selection by protein complementation assay (PCA). This new procedure allows to select specific binders by direct interaction with the target in the cytoplasm of the bacteria and consequently does not require preliminary purification of target protein. αRep binders selected by PCA or by phage display can be used to enhance expression, stability, solubility and crystallogenesis of proteins that are otherwise difficult to express, purify and/or crystallize. 相似文献
86.
Peterson CN Day S Wolfe BE Ellison AM Kolter R Pringle A 《Environmental microbiology》2008,10(9):2257-2266
The community of organisms inhabiting the water-filled leaves of the carnivorous pitcher-plant Sarracenia purpurea includes arthropods, protozoa and bacteria, and serves as a model system for studies of food web dynamics. Despite the wealth of data collected by ecologists and zoologists on this food web, very little is known about the bacterial assemblage in this microecosystem. We used terminal restriction fragment length polymorphism (T-RFLP) analysis to quantify bacterial diversity within the pitchers as a function of pitcher size, pH of the pitcher fluid and the presence of the keystone predator in this food web, larvae of the pitcher-plant mosquito Wyeomyia smithii. Results were analysed at two spatial scales: within a single bog and across three isolated bogs. Pitchers were sterile before they opened and composition of the bacterial assemblage was more variable between different bogs than within bogs. Measures of bacterial richness and diversity were greater in the presence of W. smithii and increased with increasing pitcher size. Our results suggest that fundamental ecological concepts derived from macroscopic food webs can also be used to predict the bacterial assemblages in pitcher plants. 相似文献
87.
LuTHy: a double‐readout bioluminescence‐based two‐hybrid technology for quantitative mapping of protein–protein interactions in mammalian cells 下载免费PDF全文
Philipp Trepte Sabrina Kruse Simona Kostova Sheila Hoffmann Alexander Buntru Anne Tempelmeier Christopher Secker Lisa Diez Aline Schulz Konrad Klockmeier Martina Zenkner Sabrina Golusik Kirstin Rau Sigrid Schnoegl Craig C Garner Erich E Wanker 《Molecular systems biology》2018,14(7)
Information on protein–protein interactions (PPIs) is of critical importance for studying complex biological systems and developing therapeutic strategies. Here, we present a double‐readout bioluminescence‐based two‐hybrid technology, termed LuTHy, which provides two quantitative scores in one experimental procedure when testing binary interactions. PPIs are first monitored in cells by quantification of bioluminescence resonance energy transfer (BRET) and, following cell lysis, are again quantitatively assessed by luminescence‐based co‐precipitation (LuC). The double‐readout procedure detects interactions with higher sensitivity than traditional single‐readout methods and is broadly applicable, for example, for detecting the effects of small molecules or disease‐causing mutations on PPIs. Applying LuTHy in a focused screen, we identified 42 interactions for the presynaptic chaperone CSPα, causative to adult‐onset neuronal ceroid lipofuscinosis (ANCL), a progressive neurodegenerative disease. Nearly 50% of PPIs were found to be affected when studying the effect of the disease‐causing missense mutations L115R and ?L116 in CSPα with LuTHy. Our study presents a robust, sensitive research tool with high utility for investigating the molecular mechanisms by which disease‐associated mutations impair protein activity in biological systems. 相似文献
88.
Jamshid Alaghband‐Zadeh Siroos Mehdizadeh Anne O'Farrell Judith Weisz Lucille Bitensky Joseph Chayen 《Cell biochemistry and function》1999,17(3):217-220
The previous quantitative histochemical method for measuring nitric oxide synthase (NOS) activity in tissue sections involved the loss of about 15 per cent of the NOS, presumably from the section into the reaction medium. Two changes are now described. The first is concerned with the preparation in the laboratory of the active reagent, lead ammonium citrate/acetate (LACA). The second change involves an improvement of the procedure for measuring NOS activity. The new method appears to retain all the measurable NOS activity inside the section. Copyright © 1999 John Wiley & Sons, Ltd. 相似文献
89.
Anne M. Foreman Penelope Allison Michelle Poland B. Jean Meade Oliver Wirth 《Anthrozo?s》2019,32(1):35-50
Therapy and visitation dogs are becoming more common on college campuses to provide comfort and support to students, but little attention has been given to the concerns of faculty and staff who share space with the dogs in their workplaces. The purpose of this study was to assess the perceptions of faculty and staff with regard to both the benefits and the hazards (e.g., dander, bites, fleas) and risks associated with the presence of visitation dogs in their workplaces. One hundred and thirty-eight employees who worked in buildings with resident visitation dogs completed an online survey about their perceptions of the hazards and risks of the dogs and the effects of dogs on the wellbeing of both students and employees. In general, employees perceived that the dogs presented minimal risks, and most employees believed that they can reduce stress and provide comfort to students on campus. There were a few employees, however, who reported that the dogs did not improve the work environment and conferred no benefits to the staff or students. The findings of the present survey support the mostly positive attitudes that people have for dogs in the workplace, but they also highlight a potential challenge: accommodating individuals who believe very strongly that dogs do not belong in work environments. 相似文献
90.
Anne Caroline Wiik Ernst-Otto Ropstad Ellen Bjerkås Frode Lingaas 《BMC veterinary research》2008,4(1):23