CORE--ANIMAL PHYSIOLOGY

核心--动物生理学

• 批准号: 7473189
• 负责人: MEREDITH A HAWKINS
• 金额: $ 24.03万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7473189
• 关键词: Acute-Phase Proteins; Affect; Analytical Biochemistry; Analytical Chemistry; Angiotensinogen; Animals; Aorta; Area; Biological Assay; Catheterization; Condition; Enzyme Immunoassay; Enzyme-Linked Immunosorbent Assay; Excision; Fatty acid glycerol esters; Genetic Transcription; Hexosamines; High Pressure Liquid Chromatography; Interleukin-1 beta; Leptin; Measurement; Measures; Metabolic; Metabolic syndrome; Muscle; Operative Surgical Procedures; Pathway interactions; Physiology; Plasma; Plasminogen Activator Inhibitor 1; Preparation; Protein Chemistry; Proteins; Radioimmunoassay; Serum amyloid A protein; Services; Techniques; Tissues; Tumor Necrosis Factor-alpha; Venous; Visceral; Western Blotting; cost; cytokine; glycosylation; human TNF protein; inhibitor/antagonist; programs; resistin; transcription factor

The Protein Chemistry/Animal Physiology Core will provide highly cost-effective services to the program project in the areas of protein chemistry, analytical biochemistry, and animal physiology. The Protein Chemistry component of this core will provide measurements of the amount of flux through the hexosamine pathway in tissues, and determinations of plasma levels of key proteins whose expression is likely to be regulated by the hexosamine pathway. In particular, the core will perform HPLC measurements of intracellular levels of the major hexosamine pathway products, which will indicate the degree of activation of this pathway in fat, muscle and endothelial tissue under various metabolic conditions (projects by Rossetti, Barzilai, Hawkins, Brownlee). As an additional, direct measurement of the degree of activation of this pathway, the core will use a highly specific technique ofimmtmoprecipitation and Western blotting to quantify the amount of glycosylation of a key intracellular protein, sp-1. This transcription factor, whose activation is known to be affected by its degree of glycosylation, regulates the transcription of many proteins associated with the metabolic syndrome, including many cytokines and acute phase reactants. Since this interaction is of central importance to all projects, the Core will measure plasma levels ofplasminogen activator inhibitor-1 (PAI- 1), angiotensinogen, TNF-alpha, Interleukins 1-beta and 6, Acrp30, resistin, serum amyloid A and leptin (projects by Rossetti, Barzilai, Hawkins, Shamoon), and PAI-1 levels in aortic extracts (project by Brownlee), by a variety of techniques including radioimmunoassays, ELISA assays, enzyme immunoassays and Western blotting. The Animal Physiology component of this core will provide a superlative service for animal preparation in performing indwelling arterial and venous catheterizations and surgical resections of visceral fat and aortas (projects by Rossetti, Barzilai, Brownlee).

蛋白质化学/动物生理核心将为 该计划项目在蛋白质化学，分析生物化学和动物生理学领域。该核心的蛋白质化学成分将提供通过组织中通过己糖胺途径的通量量的测量，并确定其表达可能受己糖胺途径调节的关键蛋白质的血浆水平。特别是，核心将对主要的六抗途径产物的细胞内水平进行HPLC测量，这将指示在各种代谢条件下脂肪，肌肉和内皮组织中这种途径的激活程度（Rossetti，Barzilai，Barzilai，Hawkins，Hawkins，Brownlee，Brownlee的项目）。作为该途径激活程度的另一种直接测量，核心将使用高度特异性的ImmtMoprecipitation和Western印迹技术来量化关键细胞内蛋白SP-1的糖基化量。该转录因子的活化受到其糖基化程度的影响，它调节了许多与代谢综合征相关的蛋白质的转录， 包括许多细胞因子和急性相应物。由于这种相互作用对所有项目都是至关重要的，因此核心将测量质激活剂抑制剂1的等离子体水平 （PAI-1），血管紧张素原，TNF-Alpha，Interleukins 1-beta和6，Acrp30，抗蛋白，血清淀粉样蛋白A和瘦素（Rossetti，Barzilai，Hawkins，Sharkins，Sharoon，Sharkins，Sharoon）和PAI-1水平（Project）由Brownlee），通过各种技术，包括放射免疫测定，ELISA分析，酶免疫测定和蛋白质印迹。该核心的动物生理组成部分将为动物制备提供最高的服务，以进行内脏脂肪和主动脉（Rossetti，Barzilai，Brownlee的Project）进行留置动脉和静脉导管插入术以及手术切除术。