| Characterizing the Transcriptional Effects of Endolysin Treatment on Established Biofilms of Staphylococcus aureus
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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。 ...
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| Extraction of Small Molecules from Fecal Samples and Testing of Their Activity on Microbial Physiology
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Author:
Date:
2018-04-20
[Abstract] The human body is colonized by vast communities of microbes, collectively known as microbiota, or microbiome. Although microbes colonize every surface of our bodies that is exposed to the external environment, the biggest collection of microbes colonizing humans and other mammals can be found in the gastrointestinal tract. Given the fact that the human gut is colonized by several hundred microbial species, our group hypothesized that the chemical diversity of this environment should be significant, and that many of the molecules present in that environment would have important signaling roles. Therefore, we devised a protocol to extract these molecules from human feces and test their signaling properties. Potentially bioactive extracts can be tested through addition to culture medium and ...
[摘要] 人体被巨大的微生物群体统称为微生物群体或微生物群体。尽管微生物在我们身体的每一个暴露于外部环境的表面上定殖，但人类和其他哺乳动物中最大量的微生物可以在胃肠道中找到。鉴于人类肠道已被数百种微生物物种繁殖，我们的团队假设这种环境的化学多样性应该是显着的，并且该环境中存在的许多分子将具有重要的信号传导作用。因此，我们制定了一个协议，从人类粪便中提取这些分子并测试其信号特性。可以通过添加培养基并分析细菌生长和基因表达以及其他性质来测试潜在的生物活性提取物。本文描述的方案提供了一种简便且快速的方法，用于使用肠道沙门氏菌作为模型生物体从粪便样品中提取和测试代谢物。该方案还可以适用于从其他基质如培养的哺乳动物细胞，组织，体液和无菌微生物培养物中提取小分子，并且可以针对各种微生物物种测试所得提取物。
【背景】复杂的微生物群落生活在人类和人类身上，将暴露于外部环境的每个表面都定殖。几十年来，这些社区已经获得了几个教派，其中包括正常的植物群，微生物群和最近的微生物群（Sekirov等人，2010年; ...
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| PCR-based Assay for Genome Integrity after Methyl Methanesulfonate Damage in Physcomitrella patens
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Author:
Date:
2016-10-05
[Abstract] In plant cells, genomic DNA exists in three organelles: the nucleus, chloroplast, and mitochondrion. Genomic DNA can be damaged by endogenous and exogenous factors, but the damaged DNA can be repaired by DNA repair systems. To quantify the extent of their repair activity of on individual genomic DNA, a PCR-based assay utilizing long amplicons is valuable for evaluable. This assay is based on the inhibitory effects of methyl methanesulfonate (MMS)-induced DNA damage on the amplicons. This assay is useful for assessing DNA double-strand repair pathways, such as homologous recombination repair, as it detects DNA double-strand breaks produced by MMS in vivo.
[摘要] 在植物细胞中，基因组DNA存在于三个细胞器中：核，叶绿体和线粒体。基因组DNA可以被内源和外源因子损伤，但损伤的DNA可以通过DNA修复系统修复。为了量化其对单个基因组DNA的修复活性的程度，使用长扩增子的基于PCR的测定对于可评价是有价值的。该测定基于甲磺酸甲酯（MMS）诱导的DNA损伤对扩增子的抑制作用。该测定可用于评估DNA双链修复途径，例如同源重组修复，因为其检测由MMS在体内产生的DNA双链断裂。
[背景] 基因组DNA损伤的定量可用于分析DNA修复机制。该测定利用实时PCR定量核，叶绿体和线粒体DNA拷贝数以用于长PCR产物的标准化，与由Hunter等人先前的方案相比提供更准确的定量。 （2010）。
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