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MES

MES

Company: Sigma-Aldrich
Catalog#: M3671
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Measuring CD38 Hydrolase and Cyclase Activities: 1,N6-Ethenonicotinamide Adenine Dinucleotide (ε-NAD) and Nicotinamide Guanine Dinucleotide (NGD) Fluorescence-based Methods
Author:
Date:
2018-07-20
[Abstract]  CD38 is a multifunctional enzyme involved in calcium signaling and Nicotinamide Adenine Dinucleotide (NAD+) metabolism. Through its major activity, the hydrolysis of NAD+, CD38 helps maintain the appropriate levels of this molecule for all NAD+-dependent metabolic processes to occur. Due to current advances and studies relating NAD+ decline and the development of multiple age-related conditions and diseases, CD38 gained importance in both basic science and clinical settings. The discovery and development of strategies to modulate its function and, possibly, treat diseases and improve health span put CD38 under the spotlights. Therefore, a consistent and reliable method to measure its activity and explore its use in medicine is required. We ... [摘要]  CD38是参与钙信号传导和烟酰胺腺嘌呤二核苷酸(NAD + )代谢的多功能酶。通过其主要活性,NAD + 的水解,CD38有助于维持该分子的适当水平,从而发生所有NAD + 依赖性代谢过程。由于目前有关NAD + 下降和多种年龄相关疾病和疾病发展的研究和进展,CD38在基础科学和临床环境中都具有重要意义。调整其功能,可能治疗疾病和改善健康状况的战略的发现和发展使CD38成为焦点。因此,需要一种一致且可靠的方法来测量其活性并探索其在医学中的用途。我们在这里描述了我们的小组如何测量CD38的水解酶和环化酶活性的方法,利用基于荧光的酶测定法在平板阅读器中使用1,N 6 -Ethenonicotinamide Adenine Dinucleotide(ε-NAD)进行)和烟酰胺鸟嘌呤二核苷酸(NGD)分别作为底物。

【背景】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,随着老化的进行,NAD + 水平在预期过程中降低。最近的研究表明,烟酰胺腺嘌呤二核苷酸(NAD + )的减少与年龄相关的代谢下降有关(Massudi et al。,2012)。增加NAD + 水平体内,导致延长寿命和健康跨度相关因素的活化,并改善衰老的几个生理和代谢参数(Camacho-Pereira ...

Use of Geminivirus for Delivery of CRISPR/Cas9 Components to Tobacco by Agro-infiltration
Author:
Date:
2017-04-05
[Abstract]  CRISPR/Cas9 system is a recently developed genome editing tool, and its power has been demonstrated in many organisms, including some plant species (Wang et al., 2016). In eukaryotes, the Cas9/gRNA complexes target genome sites specifically and cleave them to produce double-strand breaks (DSBs), which can be repaired by non-homologous end joining (NHEJ) pathway (Wang et al., 2016). Since NHEJ is error prone, mutations are thus generated. In plants, delivery of genome editing reagents is still challenging. In this protocol, we detail the procedure of a virus-based gRNA delivery system for CRISPR/Cas9 mediated plant genome editing (VIGE). This method offers a rapid and efficient way to deliver gRNA into plant cells, especially for those that are recalcitrant to ... [摘要]  CRISPR / Cas9系统是最近开发的基因组编辑工具,其功能已被证实在许多生物体中,包括一些植物物种(Wang等人,2016)。 在真核生物中,Cas9 / gRNA复合物特异性地靶向基因组位点并切割它们以产生双链断裂(DSB),其可以通过非同源末端连接(NHEJ)途径修复(Wang等人, 。,2016)。 由于NHEJ易出错,因此产生突变。 在植物中,基因组编辑试剂的递送仍然是挑战性的。 在本协议中,我们详细介绍了CRISPR / Cas9介导的植物基因组编辑(VIGE)的基于病毒的gRNA传递系统的过程。 该方法提供了将gRNA递送到植物细胞中的快速且有效的方式,特别是对于那些难以转基因农杆菌的方法。

已经报道了基于病毒的基因组编辑技术使用解构DNA病毒和RNA病毒(Baltes等人,2014; Ali等人,2015)。 最近,我们使用了一种完整的双因素病毒 - 卷心菜叶卷曲病毒(CaLCuV)(一种感染广西芥菜科的成员,包括花椰菜的二分酵母病毒),用于高效率 第一次(Yin等人,2015年),其主机之一的基因组编辑(Nicotiana benhamiana)首次进行基因组编辑。

59Fe Uptake Assays in Paracoccidioides Species
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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