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提供服务。这些项目提出了饮食实验，将在赠款期间每年产生2,000多个血液样本。 该核心单元将采集这些血液样本，并从血沉棕黄层制备血清、血浆和DNA。将对样本进行清点、等分、保持在-80 ℃下，并提供给项目研究者用于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内进行，并在下文列出。 将使用标准临床化学程序定量血清TSC和HDL-C（沉淀后），并通过减法定量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。使用市售试剂，采用浊度法测定ApoAI、apoB和apoE浓度，并使用市售的“ELISA”式ELISA测定Lp（a）浓度。这些检测已经在 在这个节目中使用。 非变性梯度凝胶电泳将用于使用复合梯度凝胶评估大小分辨脂蛋白中的胆固醇分布，该复合梯度凝胶能够在同一凝胶泳道中评价LDL和HDL表型。我们将测定HDL的四种组分（HDL 1a、HDL 1b、HDL z、HDL 3）和β-脂蛋白的六种组分（包括LDL的四种组分（LDL 1、LDL 2、LDL 3和LDL 4）和较大脂蛋白的两种组分（VLDL和IDL））的吸光度分数。 最后，核心单元A将负责将储存血液样本的脂质和脂蛋白表型数据和库存信息放入计算机数据文件（通过核心单元B），并将其提供给所有项目研究者。

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.