| Two Different Methods of Quantification of Oxidized Nicotinamide Adenine Dinucleotide (NAD+) and Reduced Nicotinamide Adenine Dinucleotide (NADH) Intracellular Levels: Enzymatic Coupled Cycling Assay and Ultra-performance Liquid Chromatography (UPLC)-Mass Spectrometry
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Author:
Date:
2018-07-20
[Abstract] Current studies on the age-related development of metabolic dysfunction and frailty are each day in more evidence. It is known, as aging progresses, nicotinamide adenine dinucleotide (NAD+) levels decrease in an expected physiological process. Recent studies have shown that a reduction in NAD+ is a key factor for the development of age-associated metabolic decline. Increased NAD+ levels in vivo results in activation of pro-longevity and health span-related factors. Also, it improves several physiological and metabolic parameters of aging, including muscle function, exercise capacity, glucose tolerance, and cardiac function in mouse models of natural and accelerated aging.
Given the importance of monitoring cellular NAD+ and ...
[摘要] 目前关于代谢功能障碍和虚弱的年龄相关发展的研究每天都有更多的证据。众所周知，随着衰老的进展，烟酰胺腺嘌呤二核苷酸（NAD + ）水平在预期的生理过程中降低。最近的研究表明，NAD + 的减少是与年龄相关的代谢衰退发展的关键因素。增加NAD + 水平体内导致激活寿命和健康跨度相关因素。此外，它改善了老化的几个生理和代谢参数，包括自然和加速老化的小鼠模型中的肌肉功能，运动能力，葡萄糖耐量和心脏功能。
鉴于监测细胞NAD + 和NADH水平的重要性，有一个值得信赖的方法是至关重要的。该方案的目的是在有效且广泛适用的测定中描述来自组织和细胞的NAD + 和NADH提取以及其图形和定量分析。
【背景】氧化烟酰胺腺嘌呤二核苷酸和还原型烟酰胺腺嘌呤二核苷酸（NAD + 和NADH）是重要的生物辅助因子，它们在几种合成代谢和分解代谢功能中提供和接受电子。它们参与诸如糖酵解，三羧酸循环和氧化磷酸化的反应。此外，它还作为DNA损伤修复中涉及的几种酶的底物，例如sirtuins和poly（ADP-核糖）聚合酶（PARP）（Imai和Guarente，2014; Verdin，2015; Yoshino et al。 ，2018）。
NAD + ...
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| Cell Wall-bound p-Coumaric and Ferulic Acid Analysis
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Author:
Date:
2016-10-05
[Abstract] Hydroxycinnamic acids, such as p-coumaric acid and ferulic acid, are a major class of compounds derived from the phenylpropanoid pathway. These compounds are widely conserved in plants and primarily accumulate in the secondary cell wall. They serve as important structural components that contribute to the overall strength and rigidity of plant cell walls and are also potent antioxidants valued for nutritional consumption. This protocol describes a method for analyzing hydroxycinnamic acids that are released after incubation under alkaline conditions.
[摘要] 羟基肉桂酸例如对 - 香豆酸和阿魏酸是衍生自苯丙素途径的一类主要化合物。 这些化合物在植物中广泛保守，并且主要累积在继发性细胞壁中。 它们作为重要的结构组分，有助于植物细胞壁的整体强度和刚性，并且是有价值的营养消耗的有效抗氧化剂。 该方案描述了用于分析在碱性条件下孵育后释放的羟基肉桂酸的方法。
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| Product Analysis of Starch Active Enzymes by TLC
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Author:
Date:
2015-10-20
[Abstract] Thin layer chromatography (TLC) is a useful technique for detecting the presence of monosaccharides through to oligosaccharides, though it needs to be optimized for the specific sugars that are analyzed. Here we present a method for visualizing the reaction product(s) of starch active enzymes, which can contain α-1, 4 linked and α-1, 6 linked glucose. This was first published in Molecular Microbiology (Cockburn et al., 2015). The TLC protocol is an adapted version of that published by Robyt and Mukerjea (Robyt and Mukerjea, 1994). For a summary of the products generated by starch active enzymes see the review by Hii et al. (2012).
[摘要] 薄层色谱（TLC）是用于检测单糖通过寡糖的存在的有用技术，尽管它需要针对所分析的特定糖进行优化。 在这里我们提出一种可视化的淀粉活性酶，可以包含α-1,4连接和α-1,6连接葡萄糖的反应产物的方法。 这首次发表于Molecular Microbiology（Cockburn等人，2015年）。 TLC方案是由Robyt和Mukerjea（Robyt和Mukerjea，1994）发表的适应版本。 对于由淀粉活性酶产生的产物的概述，参见Hii等人（2012）的综述。
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