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91.
Identification of SYS1 as a Host Factor Required for Shiga Toxin-Mediated Cytotoxicity in Vero Cells
Chisato Sakuma Tsuyoshi Sekizuka Makoto Kuroda Kentaro Hanada Toshiyuki Yamaji 《International journal of molecular sciences》2021,22(9)
Shiga toxin (STx) or Vero toxin is a virulence factor produced by enterohemorrhagic Escherichia coli. The toxin binds to the glycosphingolipid globotriaosylceramide (Gb3) for its entry, and causes cell death by inhibiting ribosome function. Previously, we performed a loss-of-function screen in HeLa cells using a human CRISPR knockout (KO) library and identified various host genes required for STx-induced cell death. To determine whether this library targeted to the human genome is applicable to non-human primate cells and to identify previously unrecognized factors crucial for STx-induced cell death, we herein performed a similar screen in the African green monkey kidney-derived Vero C1008 subline. Many genes relevant to metabolic enzymes and membrane trafficking were enriched, although the number of enriched genes was less than that obtained in the screening for HeLa cells. Of note, several genes that had not been enriched in the previous screening were enriched: one of these genes was SYS1, which encodes a multi-spanning membrane protein in the Golgi apparatus. In SYS1 KO Vero cells, expression of Gb3 and sphingomyelin was decreased, while that of glucosylceramide and lactosylceramide was increased. In addition, loss of SYS1 inhibited the biosynthesis of protein glycans, deformed the Golgi apparatus, and perturbed the localization of trans-Golgi network protein (TGN) 46. These results indicate that the human CRISPR KO library is applicable to Vero cell lines, and SYS1 has a widespread effect on glycan biosynthesis via regulation of intra-Golgi and endosome–TGN retrograde transports. 相似文献
92.
Alfredo Jurez-Saldivar Elizabeth Barbosa-Cabrera Edgar E. Lara-Ramírez Alma D. Paz-Gonzlez Ana V. Martínez-Vzquez Virgilio Bocanegra-García Isidro Palos Nuria E. Campillo Gildardo Rivera 《International journal of molecular sciences》2021,22(11)
Infectious diseases caused by intestinal protozoan, such as Entamoeba histolytica (E. histolytica) and Giardia lamblia (G. lamblia) are a worldwide public health issue. They affect more than 70 million people every year. They colonize intestines causing primarily diarrhea; nevertheless, these infections can lead to more serious complications. The treatment of choice, metronidazole, is in doubt due to adverse effects and resistance. Therefore, there is a need for new compounds against these parasites. In this work, a structure-based virtual screening of FDA-approved drugs was performed to identify compounds with antiprotozoal activity. The glycolytic enzyme triosephosphate isomerase, present in both E. histolytica and G. lamblia, was used as the drug target. The compounds with the best average docking score on both structures were selected for the in vitro evaluation. Three compounds, chlorhexidine, tolcapone, and imatinib, were capable of inhibit growth on G. lamblia trophozoites (0.05–4.935 μg/mL), while folic acid showed activity against E. histolytica (0.186 μg/mL) and G. lamblia (5.342 μg/mL). 相似文献
93.
Sang-Hyun Park Insu Shin Dr. Gun-Hee Kim Dr. Sung-Kyun Ko Prof. Dr. Injae Shin 《Chembiochem : a European journal of chemical biology》2021,22(24):3425-3430
A novel autophagy inhibitor, autophazole (Atz), which promoted cancer cell death via caspase activation, is described. This compound was identified from cell-based high-content screening of an imidazole library. The results showed that Atz was internalized into lysosomes of cells where it induced lysosomal membrane permeabilization (LMP). This process generated nonfunctional autolysosomes, thereby inhibiting autophagy. In addition, Atz was found to promote LMP-mediated apoptosis. Specifically, LMP induced by Atz caused release of cathepsins from lysosomes into the cytosol. Cathepsins in the cytosol cleaved Bid to generate tBid, which subsequently activated Bax to induce mitochondrial outer membrane permeabilization (MOMP). This event led to cancer cell death via caspase activation. Overall, the findings suggest that Atz will serve as a new chemical probe in efforts aimed at gaining a better understanding of the autophagic process. 相似文献
94.
Zewen Chen Zhenhua Wang Boxiang Wang Juntang Yuan Lei Huang Zengbin Yin 《International Journal of Applied Ceramic Technology》2021,18(6):2010-2019
In this study, WC-8Co cemented carbides were prepared by spark plasma sintering. When the samples sintered at 1300℃ were cooled to room temperature, the samples were sintered multiple times at 1250℃. The changes in microstructure and mechanical properties of WC-8Co cemented carbides prepared by multiple spark plasma sintering were studied. The hardness of cemented carbides increased in the first two sintering, reaching 16.5 GPa. However, the hardness decreased seriously in the last two sintering. The attenuation rates of hardness were 6.2% and 2.5% due to the abnormal coarse grains. Furthermore, the crack path along the grain boundary was almost straight, causing a decrease in the indentation fracture toughness of cemented carbides. Additionally, the grains of cemented carbides were abnormally coarsened, and the morphologies of grains became unstable due to multiple sintering. 相似文献
95.
常规粉末压片制样是一种简单、高效的绿色环保制样技术,但是应用于某些沉积物样品制备存在样片表面粗糙和粉末容易脱落的问题。实验采用高于常规的压力进行样品制备,建立了X射线荧光光谱法(XRF)分析海洋沉积物样品中包括硫、氧化钠、氧化镁、三氧化二铝、二氧化硅、五氧化二磷、氧化钾、氧化钙、二氧化钛、氧化锰、三氧化二铁、钴、镍、铜、锌、钒、铬、镓、铌、锆、钇、锶、铷、铅、钡、镧、钕和铪在内的28种主、微量组分的方法。探讨了压力为300 kN和1 600 kN时的制样效果,并尝试引入BP神经网络模型利用其非线性拟合能力校正主量组分的基体效应。结果表明,采用1 600 kN压力制备的样片,表面致密、光滑、不龟裂和不掉粉,制样重复性和测试精密度也有较大提高。以55个有证标准物质中17种组分的数据集为训练样本,建立了海洋沉积物样品中主、微量组分的遗传算法-BP神经网络预测模型。按照实验方法对各组分含量相对较低的实际样品连续测试12次,计算得方法的检出限在 0.63~634 μg/g之间;精密度试验结果表明,各组分测定值的相对标准偏差(RSD, n=7)为0.16%~25.1%。方法用于海洋沉积物实际样品分析,其分析结果与国标法的测定结果吻合,能够满足海洋沉积物样品中多种组分准确分析的要求。 相似文献
96.
The microstructure and hardness of a 2024 aluminum alloy subjected to multi-pass upsetting extrusion at ambient temperature were studied. Experimental results indicated that with the number of upsetting extrusion passes increasing, the grains of the alloy are gradually refined and the hardness increases correspondingly. After ten passes of upsetting extrusion processing, the grain size decreases to less than 200 nm in diameter and the sample maintains its original shape, while the hardness is double owing to equal-axial ultrafine grains and work hardening effect caused by large plastic deformation. 相似文献
97.
98.
Quantification of secondary structure prediction improvement using multiple alignments 总被引:9,自引:0,他引:9
Levin Jonathan M.; Pascarella Stefano; Argos Patrick; Garnier Jean 《Protein engineering, design & selection : PEDS》1993,6(8):849-854
The use of multiple sequence alignments for secondary structurepredictions is analysed. Seven different protein families, containingonly sequences of known structure, were considered to providea range of alignment and prediction conditions. Using alignmentsobtained by spatial superposition of main chain atoms in knowntertiary protein structures allowed a mean of 8% in secondarystructure prediction accuracy, when compared to those obtainedfrom the individual sequences. Substitution of these alignmentsby those determined directly from an automated sequence alignmentalgorithm showed variations in the prediction accuracy whichcorrelated with the quality of the multiple alignments and distanceof the primary sequence. Secondary structure predictions canbe reliably improved using alignments from an automatic alignmentprocedure with a mean increase of 6.87percnt;, giving an overallprediction accuracy of 68.5%, if there is a minimum of 25% sequenceidentity between all sequences in a family. 相似文献
99.
100.
Karin Yaccato Alfred Hagemeyer Andreas Lesik Anthony Volpe Henry Weinberg 《Topics in Catalysis》2004,(1):127-132
High-throughput synthesis and screening methods have been developed for the discovery of highly active lead compounds for the selective catalytic reduction as well as direct decomposition of NO in the temperature range 200–300 °C. The discovery libraries for primary screening consisted of 16 × 16 catalyst arrays on 4in. square quartz wafers. Catalysts were prepared by robotic liquid dispensing techniques and screened for catalytic activity in Symyx' scanning mass spectrometer. The scanning mass spectrometer is a fast serial screening tool that uses flat wafer catalyst surfaces, local laser heating, a scanning/sniffing nozzle and a quadrupolar mass spectrometer to compare relative catalytic activities. The feed consisted of NO/NH3 mixtures with optional O2 cofeed and Kr as the internal standard in Ar carrier gas. QMS detection allowed for tracking of H2O, N2, NO, O2, N2O and Kr. Screening protocols for catalytic materials encompassed metal precursors and carriers for supported vanadia systems, extensive doping of V2O5/TiO2, and broad screening of mixed redox metal oxides and supported base and noble metal systems. More than 500 samples could be screened in a single day. Active hits (high NO consumption accompanied by corresponding N2 production) identified in discovery libraries were re-synthesized as focus libraries for lead confirmation and further optimization. These libraries used shallower compositional gradients, for example 56 points (compositions) per ternary, with four 56-point ternaries per 4in. wafer. Broad screening ternaries were generally 8 or 15 points. The focus libraries more clearly reveal the trends and provide guidelines for secondary screening and scale-up. High conversions were achieved in scanning mass spectrometer so the scalability risk is small for the short contact time reactions. 相似文献