Biomarker Core

生物标志物核心将主要位于转化研究实验室（TRL）， 睡眠和呼吸神经生物学中心的所在地。生物标志物的形成 核心将利用现有的设施和先进的设备已经在 转化研究实验室。生物标志物核心将作为一个中央设施，收集，处理和 分析临床项目02和03产生的大量血液和尿液样本 以及大鼠组织样品和来自项目01和03的分离的人单核细胞。重要的是， 生物标志物核心将确保拟议项目收集的所有样本的质量控制。的 生物标志物核心的目标是：1）标准化研究中心之间的血液和尿液采集，2）确保 正确处理和储存样品，3）编码，记录和监控来料样品4）确保质量 控制化验，以及5）为化验提供集中设施，这些化验在内部的其他设施处不可用。 研究所。生物标志物核心将由Karen Teff博士和Miroslaw博士领导 Mackiewicz作为联合核心领导人和支持人员，包括一名高级研究技术人员和最低限度的 Teff博士实验室的一名技术人员的努力。生物标志物将分为以下几类： 氧化和保护性抗氧化剂的标志物，促炎和抗炎标志物， 粘附分子、胰岛素抵抗指数和交感神经系统活性指数。的 生物标志物核心将进行三种类型的测定：1）高压液相色谱法（HPLC）， 测量血浆和尿去甲肾上腺素和代谢物如去甲肾上腺素以及 褪黑激素，一种与睡眠相关的生物标志物，2）用于游离脂肪酸分析的分光光度法 酸和甘油三酯和3）逆转录-聚合酶链反应（RT-PCR）在大鼠舌上 组织和分离的人单核细胞。为避免资源重叠，我们会善用其他 在Penn。临床和转化研究中心实验室定期进行酶 用于测量炎性细胞因子（IL-6、TNF-α）的连接免疫吸附测定（ELISA）， 抗炎细胞因子（IL-10）和粘附分子（ICAM-1）。此外，胰岛素的标志物 需要放射免疫测定（胰岛素、瘦素、脂联素）的抵抗和敏感性将通过 宾州糖尿病研究中心放射免疫测定核心。尿异前列腺素将在 伊恩·布莱尔博士的实验室，化学教授和著名的氧化应激标志物专家。 将在Dr. Teff的实验室分析血糖。一个集中的设施将确保质量控制 和成本效益，并将支持未来的项目，建立一个样本库， 可用于未来的调查。

The Biomarker Core will be primarily located in the Translational Research Laboratory (TRL), the same building which houses the Center for Sleep and Respiratory Neurobiology. The formation of the Biomarker Core will take advantage of existing facilities and the sophisticated equipment already available within the Translational Research Laboratory. The Biomarker Core will act as a central facility to collect, process and analyze the large number of incoming blood and urine samples generated by the clinical Projects 02 and 03 as well as rat tissue samples and isolated human monocytes derived from Projects 01 and 03. Importantly, the Biomarker Core will ensure quality control of all samples collected by the proposed projects. The objectives of the Biomarker Core are to: 1) standardize blood and urine collection between sites, 2) ensure proper handling and storage of samples, 3) code, record and monitor incoming samples 4) ensure quality control of assays and 5) provide a centralized facility for assays that are not available at other facilities within the institute. The Biomarker Core will be under the leadership of Dr. Karen Teff with Dr. Miroslaw Mackiewicz as Co-Core Leader and support staff consisting of a senior research technician and minimal effort from an additional technician in Dr. Teff's laboratory. Biomarkers will fall into the following categories: markers of oxidation and protective anti-oxidants, pro-inflammatory and anti-inflammatory markers, adhesion molecules, indices of insulin resistance and indices of sympathetic nervous system activity. The Biomarker Core will conduct three types of assays: 1) high-pressure liquid chromatography (HPLC) for measurement of plasma and urinary norepinephrine and metabolites such as normetanephrine as well as melatonin, a specific sleep-related biomarker, 2) spectrophotometric methods for the analysis of free fatty acids and triglycerides and 3) Reverse Transcription - Polymerase Chain Reaction (RT-PCR) on rat tongue tissue and isolated human monocytes. To avoid duplication of resources, we will take advantages of other cores at Penn. The Clinical and Translational Research Center Laboratory routinely conducts Enzyme Linked Immunosorbent Assay (ELISA) for the measurement of the inflammatory cytokines (IL-6, TNF-a), the anti-inflammatory cytokine (IL-10) and adhesion molecules (ICAM-1). In addition, markers of insulin resistance and sensitivity requiring radioimmunoassays (insulin, leptin, adiponectin) will be measured by the Penn Diabetes Research Center Radioimmunoassay Core. Urinary isoprostanes will be measured in the laboratory of Dr. Ian Blair, Professor of Chemistry and a renowned expert in markers of oxidative stress. Plasma glucose will be analyzed in the laboratory of Dr. Teff. A centralized facility will ensure quality control and cost effectiveness and will support future projects by establishing a bank of samples that will be available for future investigation.