CORE--LIPID AND LIPOPROTEIN BIOCHEMISTRY

核心--脂质和脂蛋白生物化学

• 批准号: 7212171
• 负责人: David L. Rainwater
• 金额: $ 10.16万
• 依托单位: TEXAS BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7212171
• 关键词: apolipoproteins; biomedical facility; blood chemistry; blood lipid; blood lipoprotein; blood lipoprotein metabolism; blood preservation; cholesterol; chylomicrons; densitometry; density gradient ultracentrifugation; dietary lipid; electron microscopy; enzyme linked immunosorbent assay; gel electrophoresis; high density lipoproteins; high performance liquid chromatography; low density lipoprotein; nutrition related tag; phenotype; triglycerides; very low density lipoprotein; western blottings

The Lipid and Lipoprotein Biochemistry Core Laboratory will serve all projects on a highly interactive basis. The projects propose diet experiments that will produce more than 2,000 blood samples during each year of the grant period. This Core Unit will acquire these blood samples and prepare the serum, plasma, and DNA from the buffy coats. Samples will be inventoried, aliquotted, maintained at -80 degrees C, and made available to program investigators for determination of various cardiovascular disease-related phenotypes. Some of these determinations will be made within Core Unit A and are listed below. Standard clinical chemical procedures will be used to quantify serum TSC and HDL-C (after precipitation) and, by subtraction, VLDL+LDL-C concentrations. We will continue our long-standing participation in an external quality assurance program to ensure the accuracy of these measurements. ApoAI, apoB and apoE concentrations will be assayed using turbidometric approaches with commercially-available reagents, and Lp(a) concentrations will be assayed using a 'sandwich'-style ELISA, also commercially available. These assays are already being used in this program. Nondenaturing gradient gel electrophoresis will be used to assess cholesterol distributions among sizeresolved lipoproteins using composite gradient gels that enable evaluation of both LDL and HDL phenotypes in the same gel lane. We will determine fractional absorbance in four fractions of HDL (HDLl1a, HDL1b, HDLz, HDL3) and in six fractions of beta-lipoproteins, including four fractions ofLDL (LDL1, LDL2, LDL3, and LDL4) and two fractions of larger lipoproteins (VLDL and IDL). Finally, Core Unit A will be responsible for placing lipid and lipoprotein phenotype data and inventory information on stored blood samples into computerized data files (via Core Unit B) and making them available to all program investigators.

脂质和脂蛋白生物化学核心实验室将以高度互动的基础为所有项目提供服务。这些项目提出的饮食实验将在赠款期的每一年中产生2,000多个血液样本。 该核心单元将获取这些血液样本，并从Buffy Coats中制备血清，血浆和DNA。样品将被库存，等略性，维持在-80摄氏度，并提供给计划调查员以确定各种与心血管疾病相关的表型的确定。其中一些确定将在核心单位A内进行，并在下面列出。 标准临床化学程序将用于量化血清TSC和HDL-C（沉淀后），并通过减去VLDL+LDL-C浓度。我们将继续长期参与外部质量保证计划，以确保这些测量的准确性。将使用具有商业上可用试剂的浊度测量方法测定ApoAI，ApoB和ApoE浓度，并使用“三明治”风格的ELISA测定LP（a）浓度，也可以在市售中使用。这些测定已经在 在此程序中使用。 使用复合梯度凝胶，将使用非肾化梯度凝胶电泳来评估固定脂蛋白之间的胆固醇分布，从而可以评估同一凝胶泳道中的LDL和HDL表型。我们将确定HDL的四个部分（HDLL1A，HDL1B，HDLZ，HDL3）和六个β-脂蛋白的分数，包括LDL1，LDL1，LDL2，LDL3和LDL3和两个较大的Lipoptions（vldl）（VLDL和IDL）（VLDL和IDL）（VLDL和IDL）（VLDL和IDL）（VLDL和IDL）（VLDL和IDL）（VLDL和IDL）。 最后，核心单元A将负责将脂质和脂蛋白表型数据和库存信息放在计算机化数据文件中（通过核心单元）中，并使所有程序调查人员可用。