| Multiplication and Growth Inhibition Activity Assays for the Zoonotic Malaria Parasite, Plasmodium knowlesi
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Author:
Date:
2020-09-05
[Abstract] Malaria remains a major cause of morbidity and mortality globally. Clinical symptoms of the disease arise from the growth and multiplication of Plasmodium parasites within the blood of the host. Thus in vitro assays to determine how drug, antibody and genetic perturbations affect the growth rate of Plasmodium parasites are essential for the development of new therapeutics and improving our understanding of parasite biology. As both P. falciparum and P. knowlesi can be maintained in culture with human red blood cells, the effect of antimalarial drugs and inhibitory antibodies that target the invasion or growth capacity of Plasmodium parasites are routinely investigated by using multiplication assays or growth inhibition activity (GIA) ...
[摘要] [摘要] 疟疾仍然是全球发病率和死亡率的主要原因。该疾病的临床症状源于宿主血液中疟原虫的生长和繁殖。因此，体外测定以确定药物，抗体和遗传扰动如何影响疟原虫寄生虫的生长速率对于开发新疗法和增进我们对寄生虫生物学的理解至关重要。由于两个恶性疟原虫和P. knowlesi 可以在培养物中维持与人体红细胞，抗疟疾药物和抑制性抗体靶向的侵袭能力的影响疟原虫寄生虫 是通过使用针对这两个物种乘法测定或生长抑制测定法常规地研究。该协议给出了详细的一步一步的过程来进行基于所述寄生虫乳酸脱氢酶的活性为基础的流式细胞仪乘法测定和生长抑制活性测定法测试性中和抗体的疟原虫knowlesi 适于人类红血细胞培养物中。虽然类似测定法是用于很好地建立的恶性疟原虫，P. knowlesi 被更密切相关的所有其他人类感染性物种（帕切科等人。，2018），因此可以用作替代用于测试药物和疫苗用于其它疟疾种类，例如如间日疟原虫，它是非洲以外疟疾最广泛的病因，但尚未在实验室条件下进行培养。
[背景 ] ...
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| Analyzing the Properties of Murine Intestinal Mucins by Electrophoresis and Histology
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Author:
Date:
2017-07-20
[Abstract] Specialized secretory cells known as goblet cells in the intestine and respiratory epithelium are responsible for the secretion of mucins. Mucins are large heavily glycosylated proteins and typically have a molecular mass higher than 106 Da. These large proteins are densely substituted with short glycan chains, which have many important functional roles including determining the hydration and viscoelastic properties of the mucus gel that lines and protects the intestinal epithelium. In this protocol, we comprehensively describe the method for extraction of murine mucus and its analysis by agarose gel electrophoresis. Additionally we describe the use of High Iron Diamine-Alcian Blue, Periodic Acid Schiff’s-Alcian Blue and immune–staining methods to identify and differentiate ...
[摘要] 在肠和呼吸上皮中称为杯状细胞的专门分泌细胞负责粘蛋白的分泌。 粘蛋白是大的重糖基化蛋白质，通常具有高于10μM的分子量。 这些大蛋白质被短聚糖链密集取代，其具有许多重要的功能作用，包括测定粘液凝胶的水合和粘弹性质，其保护肠上皮。 在该方案中，我们全面地描述了通过琼脂糖凝胶电泳提取小鼠粘液的方法及其分析。 此外，我们描述了使用高铁二胺 - 阿尔星蓝，周期酸席夫氏 - 阿尔辛蓝和免疫染色方法来鉴别和区分这些粘蛋白糖蛋白上糖基化的不同状态，特别是侧重于硫酸化和唾液酸化。
【背景】一层粘液保护肠上皮，主要由粘蛋白，水，蛋白质和无机盐组成。粘液屏障的粘性和凝胶状特性使其能够物理保护和润滑粘膜，主要由粘蛋白赋予。粘蛋白是大的重糖基化蛋白质，通常具有高于10μM的分子量。然而，粘蛋白主要用O-聚糖糖装饰，其占分子量的80％。不同的位点特异性和粘蛋白特异性糖基化模式影响粘蛋白和粘液凝胶的性质。众所周知，粘蛋白糖基化在感染和疾病中发生改变（Arike等人，2017; ...
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