Forensic Age-Estimation Using Proteomics-and Genomics-Approaches

1.澄清了两种非同位素技术作为基因组学方法的应用--荧光差异显示--聚合酶链式反应用于第一次筛选，比较RT-聚合酶链式反应用于随后的确认--使我们能够识别和表征以年龄相关方式表达的新基因。利用上述方法，我们成功地在小鼠肾脏中鉴定了编码新蛋白Mpv17样蛋白M-LP的年龄依赖性表达基因，并对其分子生物学特性进行了研究，如基因结构、基因结构、剪接异构体的鉴定等。细胞生物学分析证明M-LP定位于过氧化物体膜：我们阐明了M-LP的膜靶向信号和膜的拓扑结构。此外，M-LP可能通过调节抗氧化酶基因的表达而参与活性氧的代谢。我们鉴定并鉴定了M-LP的人类同源物作为标记图像估计…更多的离子。我们成功地测定了一个年轻的年龄特异的尿糖蛋白UGL-Y的完整一级结构，它之前已经通过蛋白质组学的方法鉴定出来，UGL-Y被澄清为来自于一种基质蛋白-纤维连接蛋白的青年年龄特异的蛋白分解。对不同组织线粒体DNA(MtDNA)随年龄增长的变化特征表明，mtDNA及其转录本和mtDNA编码产物在衰老过程中含量普遍增加。我们先前已经发现人类脑下垂体中脱氧核糖核酸酶I(DNase I)的活性水平会随着年龄的变化而改变。在急性心肌梗死(AMI)发病后约3小时内，血清DNase I活性突然升高。因此，血清DNase I可作为急性心肌梗死早期诊断的新指标。经皮冠状动脉介入治疗中短暂性心肌缺血可引起血清DNase I活性显著升高。该酶的活性增加可能被证明是检测短暂性心肌缺血的敏感标志物。阐明了转录因子SPL参与的人DNase I基因表达的可能调控。较少

1. It was clarified that the application of two non isotopic techniques as a genomics approach-fluorescence differential display-PCR to the first screening and the comparative RT-PCR to the subsequent confirmation-has allowed us to identify and characterize novel genes expressed in an age-related manner.2. Using the method describe above, we succeeded in identification of an age-dependently expressed gene encoding a novel protein, Mpv17-like protein, M-LP in mouse kidney, then characterized its molecular-biological aspects such as the structure of cDNA and gene, and identification of spliced isoforms. Cell-biological analysis demonstrated that M-LP localized in peroxisomal membrane: we elucidated the peroxisomal membrane targeting signal and membrane topology of M-LP. Furthermore, M-LP may be involved in metabolism of reactive oxygen species through regulating expression of antioxidant enzyme genes.3. We identified and characterized a human homolog of the M-LP as a marker fiage-estimat … More ion.4. We succeeded in determination of the complete primary structure of a young age-specifi urinary glycoprotein, Ugl-Y, which had previously been identified by a proteomics approach Ugl-Y was clarified to be derived from a. young age-specific proteolysis of one of the matri proteins, fibronection.5. Characterization of age-related alterations in the mitochondrial DNA (mtDNA) of various tissues revealed that mtDNA, its transcripts and mtDNA encoded products generally increase in content during the course of aging.6. We have previously found levels of deoxyribonuclease I (DNase I) activity in human pituitary gland to alter in an age-related manner. An abrupt elevation of serum DNase I activity was observed within approximately 3 hours of the onset of acute myocardial infarction (AMI. Therefore, serum DNase I could be used as a new diagnostic marker for the early detection of AMI.7. Transient myocardial ischemia during percutaneous coronary intervention induced a significant elevation of serum DNase I activity. Increased activity of the enzyme may prove useful as a sensitive marker for detection of transient myocardial ischemia.8. Possible regulation of human DNase I gene expression, in which transcription factor Spl is involved, was elucidated. Less