CORE A -LIPID AND LIPOPROTEIN BIOCHEMISTRY

脂质和脂蛋白生物化学核心实验室将在高度互动的基础上为所有项目服务。这些项目提出了饮食实验，在资助期的每一年将产生2000多个血液样本。这个核心单位将获取这些血液样本，并从巴菲大衣中制备血清、血浆和DNA。样本将被清点，等分，保持在-80摄氏度，并提供给程序调查人员，以确定各种心血管疾病相关表型。其中一些决定将在核心单元A内作出，如下所示。 标准的临床化学程序将用于量化血清总胆固醇和高密度脂蛋白胆固醇(沉淀后)，并通过减去极低密度脂蛋白-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。载脂蛋白AI、载脂蛋白B和载脂蛋白E的浓度将通过使用商用试剂的浊度法进行检测，而Lp(A)的浓度将使用同样可以从市场上获得的夹心式酶联免疫吸附试验进行检测。这些化验已经在 在此程序中使用。 非变性梯度凝胶电泳法将使用复合梯度凝胶，在同一凝胶通道中评估低密度脂蛋白和高密度脂蛋白的表型，以评估胆固醇在大分子脂蛋白中的分布。我们将测定四种高密度脂蛋白(HDL1a、HDL1b、HDLz、HDL3)和六种β-脂蛋白的吸光度，包括四种低密度脂蛋白(低密度脂蛋白1、低密度脂蛋白2、低密度脂蛋白3和低密度脂蛋白4)和两种较大的脂蛋白(极低密度脂蛋白和低密度脂蛋白)。 最后，核心单位A将负责将储存的血液样本上的脂类和脂蛋白表型数据和库存信息放入计算机化的数据文件中(通过核心单位B)，并将其提供给所有项目调查人员。

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.