DNA, PROTEINS & CARBON TETRACHLORIDE HEPATOTOXICITY

该项目的长期目标是了解如何 引发化学物质的自由基遗传物质和/或其 通过研究肝毒素CC14对肝脏DNA的影响来表达 和不同的核(N)蛋白。观察到的影响将是 与已知的肝脏明显不同的反应相关 从不同的动物物种或品系到CC14的作用(坏死， 肝硬变、癌症)(从现在起在文字上称为“肝脏反应” 至CC14“)。 本项目的具体目标是：a)核实是否有 14CC14与DNA或TO的共价结合(CB)相关性 不同n-蛋白质组分与已知的“肝脏对CC14的反应” 不同的动物种类或品系。A)启动结构研究 CC13自由基与DNA碱基反应生成的加合物 和氨基酸。C)核实以下各项之间是否存在相关性 CC1在体内或体外促进脂质过氧化(LP)的能力 (肝脏切片、微粒体和细胞核)和已知的“肝脏对 不同动物种类或品系的CC14“。d)验证是否 LP过程中产生的花生四烯酸降解产物(例如，4 )能够在体内或体外与DNA或n- 蛋白质。E)研究CC13诱导的DNA碱基和氨基酸的变化 不能归因于CB或LP的酸(例如，形成交联键， 细分)。 为了实现这些目标，我们计划：a)使高度纯化的DNA和 不同的n-蛋白质组分(例如，组蛋白、酸性等)并向 确定CB对他们的影响程度。B)试图识别 用高效液相色谱法测定改变的碱基或核苷和氨基酸 (UV/14C/荧光示波电测法)和/或GLC/MS 不同的实验条件。E)确定CC14是否促进 核Lp由乙烷或丙二醛或4 羟基壬烯醛生产。 该项目对健康的主要影响：实现更好的 对自由基致DNA和n-蛋白质改变的认识 以及它们在急性和长期化学n=诱导中的潜在作用 肝脏受损。结果可能会使我们对其作用机制有一个深入的了解。 具有弱致突变性或无致突变性的致癌物，如CC14。

The long term objective of the project are to gain knowledge baout how free radical causing chemicals later genetic material and/or its expression, by studying the effects of the hepatotoxin CC14 on liver DNA and different nuclear (n) proteins. The observed effects would be correlated with the already known markedly different response of livers from different animal species or strains to CC14 action (necrosis, cirrhosis, cancer) (from now on in text referred as the "liver response to CC14"). The present project specific aims are: a) To verify whether there is a correlation between the covalent binding (CB) of 14CC14 to DNA or to different n-protein fractions and the known "liver response to CC14" of different animal species or strains. a) To initiate structural studies of the adducts formed when the CC13 free radicals react with DNA bases and amino acids. c) To verify whether there is a correlation between CC1 ability to promote lipid peroxidation (LP) "in vivo" or "in vitro" (liver slices, micorsomes and nuclei) and the known "liver response to CC14" of different animal species or strains. d) To verify whether arachidonic acid degradation products produced during LP (e.g., 4 hydroxynonenal) are able to CB "in vivo" or "in vitro" to DNA or n- proteins. e) To study CC13 induced alterations in DNA bases and amino acids not attributable to either CB or LP (e.g., cross links formation, breakdown). To achieve these goals we plan: a) To islate highly purified DNA and different n-protein fractions (e.g., histones, acidic, etc.) and to establish the extent of CB to them. b) To attempt identification of altered bases or nucleosides and amino acids by HPLC (UV/14C/fluoroscence.electrometric detection) and/or by GLC/MS under different experimental conditions. e) To establish whether CC14 promotes nuclear LP as determined by ethanepenthane or malondialdehyde or 4 hydroxynonenal production. Major Health Implications of The Project: To achieve a better understanding of free radical induced alterations in DNA and n-proteins and their potential role in acute and long term chemically n=induced liver damage. Results might give an insight to mechanisms of action of liver carcinogens of weak or null mutagenic nature as is CC14.