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LB Broth

LB肉汤

Company: Thermo Fisher Scientific
Catalog#: 10855001
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Adhesion of Enteroaggregative E. coli Strains to HEK293 Cells
Author:
Date:
2018-04-20
[Abstract]  Enteroaggregative Escherichia coli (EAEC) is a recognized cause of acute diarrhea among both children and adults worldwide. EAEC strains are characterized by the presence of aggregative adherence fimbriae (AAF), which play a key role in pathogenesis by mediating attachment to the intestinal mucosa and by triggering host inflammatory responses. The aggregative adherence fimbria II (AAF/II) is the most important adherence factor of EAEC prototype strain 042 (EAEC042) to intestinal cells. Multiple receptors for AAF/II on epithelial cells have been identified including the transmembrane signaling mucin Muc1. This protocol describes a method to measure adherence of EAEC strains to HEK293 cells expressing the Muc1 glycoprotein. [摘要]  肠道集聚性大肠杆菌(EAEC)是全球儿童和成人急性腹泻的公认原因。 EAEC菌株的特征在于存在聚集粘附菌毛(AAF),其通过介导与肠粘膜的附着和通过引发宿主炎症反应而在发病机制中起关键作用。 聚合粘附菌毛II(AAF / II)是EAEC原型菌株042(EAEC042)对肠细胞最重要的粘附因子。 已经鉴定了上皮细胞上AAF / II的多种受体,包括跨膜信号传导粘蛋白Muc1。 该协议描述了测量EAEC菌株对表达Muc1糖蛋白的HEK293细胞的依从性的方法。

【背景】EAEC是世界范围内地方性和流行性腹泻病的重要原因。尽管发展中国家儿童腹泻最常见,但EAEC还与免疫受损成人腹泻,旅行者和工业化国家的食源性疾病有关,例如由志贺毒素(Stx)2a型产生的大致致命爆发2011年在北欧的血清型O104:H4的EAEC菌株(Harrington等人,2006; Rasko等人,2011)。 EAEC发病机制由生物体粘附肠细胞,产生肠毒素和细胞毒素并最终诱导炎症的能力决定(Harrington等,2006)。 EAEC对肠细胞的依从性由AAF菌毛粘附素介导(Czeczulin等人,1997)。迄今为止,已经描述了至少5种AAF菌毛的变体,全部编码在范围为55至65MDa的毒力质粒中(Jonsson等人,2015)。 ...

Induction of Connexin-hemichannel Opening
Author:
Date:
2014-09-05
[Abstract]  Connexins (Cxs) are integral membrane proteins of vertebrates that associate to form hexameric transmembrane channels, named hemichannels. Twenty-one Cx types have been described, which are named according to their molecular weight. Cxs are expressed in many cell types, e.g. epithelial cells, astrocytes and immune cells. Hemichannels allow the passage of molecules of up to 1-2 kDa along the concentration gradient. When surface-exposed, hemichannels mediate the exchange of molecules between the cytosol and the extracellular space. Hemichannels are closed by default, but several cues inducing their opening have been described, e.g. a drop in the extracellular Ca2+ concentration (Evans et al., 2006) or infection with enteric pathogens (Puhar et al. ... [摘要]  连接蛋白(Cxs)是脊椎动物的内在膜蛋白,其结合形成六聚体跨膜通道,称为半通道。已经描述了21种Cx类型,其根据它们的分子量命名。 Cxs在许多细胞类型中表达,例如上皮细胞,星形胶质细胞和免疫细胞。半通道允许高达1-2kDa的分子沿浓度梯度通过。当表面暴露时,半通道介导细胞溶质和细胞外空间之间的分子交换。半通道被默认关闭,但是已经描述了诱导其开放的几个线索,例如,细胞外Ca 2+浓度的下降(​​Evans等人, 2006);或用肠道病原体感染(Puhar et al。,2013; Tran Van Nhieu等人,2003)。该方案与上皮细胞,特别是与极化和非极化肠上皮TC7细胞和用Cx26或Cx43稳定转染的Hela细胞(Paemeleire等人,2000)一起使用。然而,它可能可用于其他Cx表达细胞类型。是否半通道是开放的可以通过电生理学或通过测量半通道可渗透分子(例如,ATP)在细胞外介质中的释放或半通道可渗透的质膜不渗透分子的摄取来确定[例如,荧光染料溴化乙锭 - 参见相关协议"通过Connexin Hemichannels进行染料吸收实验"(Puhar和Sansonetti,2014 )]。

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