| Live-cell Imaging of Neisseria meningitidis Microcolony Dispersal Induced by Lactate or Other Molecules
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Author:
Date:
2018-01-20
[Abstract] To efficiently colonize the nasopharyngeal epithelium, the human restricted pathogen Neisseria meningitidis follows a multistep adhesion cascade. First, the bacteria adhere to host cells and aggregate into spherical shaped structures called microcolonies. Several hours later, single bacteria start dispersing from the microcolonies and form a monolayer on top of the host cells. Once in proximity to host cells meningococci can adhere tightly to the epithelial surface or become internalized. This can eventually result in invasion of the mucosal surfaces and gain access to the bloodstream, causing a life-threatening disease. Lactate, a metabolite derived from human epithelial cells, has been previously shown to induce rapid dispersal of N. meningitidis from microcolonies. ...
[摘要] 为了有效地定居鼻咽上皮，人类限制性病原体脑膜炎奈瑟氏球菌遵循多步粘附级联。首先，细菌粘附到宿主细胞并聚集成称为微菌落的球形结构。几个小时后，单细菌开始从微菌落分散并在宿主细胞上形成单层。一旦接近宿主细胞，脑膜炎球菌可紧密地粘附在上皮表面或内化。这最终可能导致粘膜表面的侵入并进入血液，导致危及生命的疾病。乳酸是一种来源于人类上皮细胞的代谢物，之前已被证明能诱导N的快速分散。 meningitidis 来自微菌落。在这里，我们描述基于活细胞成像的宿主细胞自由方法来检查宿主来源的乳酸对N的时间的影响。 meningitides microcolony扩散。虽然在这个协议中我们使用乳酸盐，它可以很容易地修改，以测试其他分子的影响。
【背景】ñ。脑膜炎是引起败血症和/或脑膜炎的专性人类病原体。鼻咽上皮的初始附着和随后形成的小菌落是建立感染的第一步。为了导致疾病N。脑膜炎奈瑟球必须穿过鼻咽粘膜上皮屏障，其天然储库，并进入血液（Stephens，2009; ...
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| Quantification of 2-Hydroxyglutarate Enantiomers by Liquid Chromatography-mass Spectrometry
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Author:
Date:
2016-08-20
[Abstract] Two enantiomers of 2-hydroxyglutarate (2HG), L (L2HG) and D (D2HG), are metabolites of unknown function in mammalian cells that were initially associated with separate and rare inborn errors of metabolism resulting in increased urinary excretion of 2HG linked to neurological deficits in children (Chalmers et al., 1980; Duran et al., 1980; Kranendijk et al., 2012). More recently, investigators have shown that D2HG is produced by mutant isocitrate dehydrogenase enzymes associated with a variety of human malignancies, such as acute myeloid leukemia, glioblastoma multiforme, and cholangiocarcinoma (Cairns and Mak, 2013; Dang et al., 2009; Ward et al., 2010). By contrast, we and others have shown that L2HG accumulates in response to cellular ...
[摘要] 2-羟基戊二酸（2HG），L（L2HG）和D（D2HG）的两种对映异构体是哺乳动物细胞中未知功能的代谢物，其最初与分开的和罕见的代谢的先天性错误相关，导致与神经系统相关的2HG的尿排泄增加儿童中的缺陷（Chalmers等人，1980; Duran等人，1980; Kranendijk等人，2012）。最近，研究者已经表明，D2HG由与多种人类恶性肿瘤相关的突变体异柠檬酸脱氢酶产生，所述人类恶性肿瘤例如急性骨髓性白血病，多形性成胶质细胞瘤和胆管癌（Cairns和Mak，2013; Dang等人，2009; Ward等人，2010）。相比之下，我们和其他人已经显示L2HG响应于细胞还原应激物如缺氧，缺氧诱导因子的激活和线粒体电子传递链缺陷而累积（Oldham等人，2015; Reinecke 2011; Intlekofer等人，2015; ...
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| Glucose Production Assay in Primary Mouse Hepatocytes
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Author:
Date:
2012-11-05
[Abstract] Hepatic glucose production is a primary determinant of fasting hyperglycemia in type 2 diabetic patients. Glucagon-cAMP-PKA pathway increases, but insulin-PI3 kinase-Akt pathway suppresses glucose production. This assay aims to evaluate the ability of isolated mouse hepatocytes to release newly synthesized glucose mainly from lactate and pyruvate as the substrates (i.e. gluconeogenesis) under basal, cAMP-, or cAMP plus insulin-treated condition.
[摘要] 肝葡萄糖产生是2型糖尿病患者的空腹高血糖的主要决定因素。 胰高血糖素-cAMP-PKA通路增加，但胰岛素-PI3激酶-akt通路抑制葡萄糖产生。 该测定旨在评价分离的小鼠肝细胞主要从作为在基础，cAMP或cAMP加胰岛素治疗的条件下的底物（即，糖原异生）的乳酸盐和丙酮酸释放新合成的葡萄糖的能力。
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