Forensic Age-Estimation Using Proteomics-and Genomics-Approaches

1.阐明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PCR用于首次筛选和比较RT-PCR用于随后的确认-使我们能够鉴定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一个编码新蛋白质Mpv 17-like protein（M-LP）的年龄依赖性表达基因，然后对其分子生物学方面进行了表征，如cDNA和基因的结构，以及剪接异构体的鉴定。细胞生物学分析表明，M-LP定位于过氧化物酶体膜：我们阐明了过氧化物酶体膜靶向信号和膜拓扑结构的M-LP。此外，M-LP可能通过调节抗氧化酶基因的表达参与活性氧代谢.我们鉴定并鉴定了M-LP的人类同源物作为标记物， ...更多信息 离子。我们成功地确定了一个年轻的年龄特异性尿糖蛋白Ugl-Y的完整的一级结构，这是以前通过蛋白质组学方法鉴定的。年轻的年龄特异性蛋白质水解的基质蛋白之一，纤连蛋白。对不同组织线粒体DNA（mtDNA）中与年龄相关的变化的表征表明，mtDNA、其转录本和mtDNA编码产物的含量在衰老过程中普遍增加.我们先前发现人类脑垂体中脱氧核糖核酸酶I（DNase I）活性水平以与年龄相关的方式改变。在急性心肌梗死（AMI）发病后约3小时内观察到血清DNA酶I活性突然升高。因此，血清DNA酶I可作为AMI早期诊断的一个新指标.经皮冠状动脉介入治疗期间短暂心肌缺血引起血清DNase I活性显著升高。该酶活性的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DNA酶I基因表达的可能调控。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