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MicroAmp® Optical Adhesive Film

MicroAmp ®光学胶粘膜

Company: Thermo Fisher Scientific
Catalog#: 4311971
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Long-term in vitro Culture of Cryptosporidium parvum
Author:
Date:
2018-08-05
[Abstract]  Continuous in vitro growth of Cryptosporidium parvum has proved difficult and conventional in vitro culture techniques result in short-term (2-5 days) growth of the parasite resulting in thin-walled oocysts that fail to propagate using in vitro cultures, and do not produce an active infection using immunosuppressed or immunodeficient mouse models (Arrowood, 2002). Here we describe the use of hollow fiber bioreactors (HFB) that simulate in vivo conditions by providing oxygen and nutrients to host intestinal cells from the basal surface and permit the establishment of a low redox, high nutrient environment on the apical surface. When inoculated with 105 C. parvum (Iowa isolate) oocysts the bioreactor produced 108 ... [摘要]  Cryptosporidium parvum 的连续体外生长已证明是困难的,并且常规体外培养技术导致短期(2-5天)生长寄生虫导致薄壁卵囊不能使用体外培养物繁殖,并且不使用免疫抑制或免疫缺陷小鼠模型产生活跃感染(Arrowood,2002)。在这里,我们描述了中空纤维生物反应器(HFB)的使用,通过提供氧气和营养物质从基础表面宿主肠细胞模拟体内条件,并允许建立低氧化还原,高营养环境顶面。当接种10 5 C时。 parvum (爱荷华州分离物)卵囊生物反应器在14天后每ml产生10个 8 卵囊(20ml额外毛细血管体积),并保持2年以上。使用TCR-α免疫缺陷小鼠模型的体内感染性研究显示,在6,12和18个月时从生物反应器产生的卵囊与用于启动培养的亲本Iowa分离物无法区分。 HFB产生的卵囊具有与亲本爱荷华分离物类似的百分比分析。

【背景】 Cryptosporidium parvum 是人和其他哺乳动物肠道的细胞内专性寄生虫,导致急性腹泻。该疾病在免疫功能正常的个体中是自限性的,然而,在免疫功能低下的成人和幼儿中,该疾病可能危及生命(Kotloff,2017)。它是经济资源低的国家中三种被诊断出的儿童肠道疾病之一(Kotloff et al。,2013; Sow et ...

Characterizing the Transcriptional Effects of Endolysin Treatment on Established Biofilms of Staphylococcus aureus
Author:
Date:
2018-06-20
[Abstract]  Biofilms are the most common lifestyle of bacteria in both natural and human environments. The organized structure of these multicellular communities generally protects bacterial cells from external challenges, thereby enhancing their ability to survive treatment with antibiotics or disinfectants. For this reason, the search for new antibiofilm strategies is an active field of study. In this context, bacteriophages (viruses that infect bacteria) and their derived proteins have been proposed as promising alternatives for eliminating biofilms. For instance, endolysins can degrade peptidoglycan and, ultimately, lyse the target bacterial cells. However, it is important to characterize the responses of bacterial cells exposed to these compounds in order to improve the design of phage-based ... [摘要]  生物膜是自然和人类环境中最常见的细菌生活方式。这些多细胞社区的有组织结构通常保护细菌细胞免受外部挑战,从而增强其抗生素或消毒剂治疗的生存能力。为此,寻找新的抗菌膜策略是一个积极的研究领域。在这种情况下,已提出噬菌体(感染细菌的病毒)及其衍生蛋白作为消除生物膜的有希望的替代物。例如,内溶素可降解肽聚糖,并最终裂解靶细菌细胞。然而,表征暴露于这些化合物的细菌细胞的反应以改进基于噬菌体的抗微生物策略的设计是重要的。

如以前在Fernández等人(2017)中所描述的,开发该协议以检查暴露于内溶素处理的金黄色葡萄球菌生物膜细胞的转录反应。然而,它可能随后适用于分析其他微生物对不同抗菌剂的反应。

【背景】越来越清楚的是,亚抑制剂量的抗菌剂可能对目标微生物的不同表型具有调节作用,包括生物膜形成,代谢或毒力。因此,研究新化合物对低浓度靶细胞的潜在影响应该是发展过程的一部分。事实上,引发毒力因子或抗生素耐药决定簇产生的非常有效的抗菌剂可能不是治疗应用的良好候选者。另一方面,考虑到生物膜和浮游细胞之间的生理差异,应该对生物膜形成细胞分析新抗生物膜剂的作用似乎是合乎逻辑的。在这里,我们描述了一种协议,用于分析生物膜细胞在亚抑制浓度的内抑素浓度下的转录反应,噬菌体来源的蛋白质作为生物膜去除剂展现出巨大的前景。因此,通过RNA-seq将内溶素处理的细胞的转录组与对照细胞进行比较,并且后来通过RT-qPCR证实了所选基因的差异表达。 ...

Detection and Analysis of Circular RNAs by RT-PCR
Author:
Date:
2018-03-20
[Abstract]  Gene expression in eukaryotic cells is tightly regulated at the transcriptional and posttranscriptional levels. Posttranscriptional processes, including pre-mRNA splicing, mRNA export, mRNA turnover, and mRNA translation, are controlled by RNA-binding proteins (RBPs) and noncoding (nc)RNAs. The vast family of ncRNAs comprises diverse regulatory RNAs, such as microRNAs and long noncoding (lnc)RNAs, but also the poorly explored class of circular (circ)RNAs. Although first discovered more than three decades ago by electron microscopy, only the advent of high-throughput RNA-sequencing (RNA-seq) and the development of innovative bioinformatic pipelines have begun to allow the systematic identification of circRNAs (Szabo and Salzman, 2016; Panda et al., 2017b; Panda et al., ... [摘要]  真核细胞中的基因表达在转录和转录后水平受到严格调控。 mRNA转录,mRNA转录和mRNA翻译等后转录过程由RNA结合蛋白(RBPs)和非编码(nc)RNAs控制。大量的ncRNA家族包含多种调控RNA,如microRNAs和长的非编码(lnc)RNAs,但也是探索不足的一类环状RNAs。虽然三十多年前电子显微镜首次发现,但只有高通量RNA测序(RNA-seq)的出现和创新生物信息学管道的开发已经开始允许系统鉴定circRNA(Szabo和Salzman,2016;熊猫,2017b;熊猫等,2017c)。然而,通过RNA测序鉴定的真正的circRNA的验证需要其他分子生物学技术,包括常规或定量(q)聚合酶链反应(PCR)和Northern印迹分析(Jeck和Sharpless,2014)的逆转录(RT)。使用不同引物的环状RNA的RT-qPCR分析已被广泛用于检测,验证和有时定量circRNA(Abdelmohsen等人,2015和2017; Panda等人, ,2017b)。如在此详述的,设计为跨越循环RNA后接连接序列的分歧引物可以特异性扩增circRNA而不是对应的线性RNA。总之,使用不同引物的RT-PCR分析允许直接检测和定量circRNA。

【背景】CircRNAs是共价闭合的,缺少5'或3'末端的单链RNA。虽然它们的起源知之甚少,但它们可以通过称为反向剪接的过程从前体mRNA产生(Panda等人,2017d; ...

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