| Estimation of the Minimum Number of Replication Origins Per Chromosome in any Organism
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Author:
Date:
2020-10-20
[Abstract] Eukaryote nuclear genomes predominantly replicate through multiple replication origins. The number of replication origins activated per chromosome during the S-phase duration may vary according to many factors, but the predominant one is replication stress. Several studies have applied different approaches to estimate the number and map the positions of the replication origins in various organisms. However, without a parameter to restrict the minimum of necessary origins, less sensitive techniques may suggest conflicting results. The estimation of the minimum number of replication origins (MO) per chromosome is an innovative method that allows the establishment of a threshold, which serves as a parameter for genomic approaches that map origins. For this, the MO can be easily ...
[摘要] [摘要] 比率可能因多种因素而变化，但最主要的因素是复制应力。一些研究应用了不同的方法来估计复制源在不同生物体中的数量和位置。然而，如果没有一个参数来限制必要起源的最小值，那么不太敏感的技术可能会产生相互矛盾的结果。估计每个染色体的最小复制源数量（MO）是一种创新的方法，它允许建立一个阈值，作为绘制起源的基因组方法的参数。为此，MO可以很容易地通过一个公式得到，这个公式需要作为参数：染色体大小、S期持续时间和复制率。在基因组数据库（如NCBI）中可以获得任何生物体的染色体大小，通过监测DNA复制来估计S期的持续时间，并通过DNA组合方法获得复制率。 提供了一种简单、快速的估算MO的方法一个新的方法学框架来协助研究任何有机体。
关键词： DNA复制，复制来源，复制率，S期持续时间，染色体大小
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| Immunohistochemistry of Kidney a-SMA, Collagen 1, and Collagen 3, in A Novel Mouse Model of Reno-cardiac Syndrome
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Author:
Date:
2020-09-20
[Abstract] Cardiorenal syndrome defines a synergistic pathology of the heart and kidneys where failure of one organ causes failure in the other. The incidence of cardiovascular mortality caused by this syndrome, is 20 fold higher in the end stage renal disease (ESRD) population compared to the population as a whole thus necessitating the need for improved therapeutic strategies to combat reno-cardiac pathologies.
Murine in vivo models play a major role in such research permitting precise genetic modification thus reducing miscellany, however presently there is no steadfast model of reno-cardiac syndrome in the most common genetically modified mouse strain, the C57BL/6 mouse. In this study we have modified an established model of chronic renal disease using adenine diet and ...
[摘要] [摘要 ] 心肾综合征定义了心脏和肾脏的协同病理，其中一个器官的衰竭导致另一个器官的衰竭。与整个人群相比，该综合征导致的心血管疾病死亡率在终末期肾脏病（ESRD）人群中要高出20倍，因此有必要改善治疗策略以应对肾病。
小鼠体内模型在允许精确基因修饰从而减少杂项的研究中起主要作用，但是目前在最常见的基因修饰小鼠品系C57BL / 6小鼠中还没有稳定的雷诺-心脏综合征模型。在这项研究中，我们使用腺嘌呤饮食修改了已建立的慢性肾脏疾病模型，并扩展了在C57BL / 6小鼠中实现慢性肾功能衰竭和随之而来的肾心脏综合征的相关病理。
使八周大的雄性C57BL / 6小鼠适应7天，然后给予0.15％腺嘌呤饮食或对照饮食20周，此后终止实验，收集血液，尿液和器官并进行生化和免疫组织化学分析。
施用0.15％的腺嘌呤饮食会导致进行性肾功能衰竭，从而导致肾性心脏病综合征，这可通过心体重比显着增加来证实（P <0.0001）。血液生化表明，用腺嘌呤喂养的小鼠血清肌酐，尿素含量显着增加（P <0.0001），肾小球滤过率显着降低（P <0.05），而肾脏的α-SMA，胶原蛋白1和胶原蛋白3免疫组化显示严重的纤维化。
我们提出了一种新型的腺嘌呤饮食方案，该方案在C57BL / ...
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| SMART (Single Molecule Analysis of Resection Tracks) Technique for Assessing DNA end-Resection in Response to DNA Damage
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Author:
Date:
2020-08-05
[Abstract] DNA double strand breaks (DSBs) are among the most toxic lesions affecting genome integrity. DSBs are mainly repaired through non-homologous end joining (NHEJ) and homologous recombination (HR). A crucial step of the HR process is the generation, through DNA end-resection, of a long 3′ single-strand DNA stretch, necessary to prime DNA synthesis using a homologous region as a template, following DNA strand invasion. DNA end resection inhibits NHEJ and triggers homology-directed DSB repair, ultimately guaranteeing a faithful DNA repair. Established methods to evaluate the DNA end-resection process are the immunofluorescence analysis of the phospho-S4/8 RPA32 protein foci, a marker of DNA end-resection, or of the phospho-S4/8 RPA32 protein levels by Western blot. Recently, the Single ...
[摘要] [摘要] DNA双链断裂（dsb）是影响基因组完整性的最具毒性的损伤之一。dsb主要通过非同源末端连接（NHEJ）和同源重组（HR）进行修复。HR过程的一个关键步骤是通过DNA末端切除，产生一个长的3′单链DNA链，这是在DNA链入侵后，以同源区域为模板进行DNA合成所必需的。DNA末端切除抑制NHEJ并触发同源定向的DSB修复，最终保证DNA的可靠修复。已建立的评价DNA末端切除过程的方法是免疫荧光法分析磷酸化S4/8rpa32蛋白病灶（DNA末端切除的标志物）或磷酸化S4/8rpa32蛋白水平。近年来，切除轨迹单分子分析（SMART）被认为是一种可靠的方法，可以通过免疫荧光法观察S期特异性DNA损伤剂（如喜树碱）处理细胞后产生的长3′单链DNA尾。然后，通过图像分析软件（如Photoshop）测量DNA束长度，评价DNA末端切除机的处理能力。DNA纤维的制备是在非变性条件下进行的，因此免疫荧光只检测DSB处理产生的特定的3′单链DNA尾。
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