| Adhesion and Invasion Assay Procedure Using Caco-2 Cells for Listeria monocytogenes
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Author:
Date:
2017-05-05
[Abstract] Listeria monocytogenes is an important Gram-positive foodborne pathogen that is a particular problem in ready-to-eat food. It has an ability to survive in harsh conditions like refrigeration temperatures and high salt concentrations and is known to cross intestinal, placental and blood-brain barriers. Several cancerous cell lines like cervical, liver, dendritic, intestinal and macrophages have been used to study in vitro propagation and survival of listeria in human cells. Human intestinal epithelial cells have been used to study how listeria crosses the intestinal barrier and cause infection. The protocol in this articles describes the procedures to grow Caco-2 cells, maintain cells and use them for adhesion and invasion assays. During adhesion assay the cells are ...
[摘要] 单核细胞增生利斯特氏菌是一种重要的革兰氏阳性食源性病原体，是即食食品中的一个特殊问题。它具有在诸如制冷温度和高盐浓度的恶劣条件下生存的能力，并且已知可以穿过肠，胎盘和血脑屏障。已经使用了诸如子宫颈，肝脏，树突状细胞，肠和巨噬细胞的几种癌细胞系来研究人细胞中李斯特菌的体外扩增和存活。人肠上皮细胞已被用于研究李斯特菌如何穿过肠屏障并引起感染。本文中的方案描述了生长Caco-2细胞的过程，维持细胞并将其用于粘附和侵袭测定。在粘附测定期间，将细胞与李斯特菌孵育30分钟，但是在侵袭测定中，细胞生长在感染后的几个时间点被停止以监测细胞中李斯特菌的生长和存活率。
背景 ...
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| 59Fe Uptake Assays in Paracoccidioides Species
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Author:
Date:
2016-09-20
[Abstract] Iron is an essential micronutrient required for virtually all organisms. This fact is related to the ability of the transition metal to exist in two oxidation states, the reduced ferrous (Fe2+) and the oxidized ferric (Fe3+). Given the relative availability of aqueous iron (the element which constitutes ~5% of the earth’s crust) one is not surprised that iron is the most common prosthetic element in biology. Usually, fungi can uptake iron through receptor-mediated internalization of a siderophore or heme, and/or reductive iron assimilation (RIA) (Kosman, 2013). In this way, the uptake of iron in the absence or presence of the reducing agent ascorbic acid can be investigated by 59Fe uptake assays, as previously described (Eide et al., 1992). In the ...
[摘要] 铁是几乎所有生物体所必需的微量营养素。这一事实涉及过渡金属以两种氧化态存在的能力，即还原的亚铁（Fe 2+）和氧化的铁（Fe 3++）。考虑到铁水（构成地壳的〜5％的元素）的相对可用性，人们不惊讶的是，铁是生物学中最常见的假体元素。通常，真菌可以通过受体介导的铁载体或血红素的内化，和/或还原铁同化（RIA）吸收铁（Kosman，2013）。以这种方式，如上所述，可以通过59 Fe吸收测定来研究在还原剂抗坏血酸存在或不存在下铁的吸收（Eide等人）。 ，1992）。在抗坏血酸存在下，研究还原非依赖性59 Fe吸收途径。另一方面，在不存在抗坏血酸的情况下，刺激还原依赖性59 Fe吸收途径。使用这种策略用于人类致病真菌Paracoccidioides物种，结果表明，在没有抗坏血酸的情况下，通过 01的铁摄取是低的，不同于在 > Pb 18。这些结果表明，只有在Fe 18中，铁摄取路径与铁还原酶偶联（Bailão等人，2015）。在该方案中，我们描述了如何在Paracoccidioides物种中进行 59 Fe摄取测定。
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