Biomarker Core

Biomarker核心将主要设在翻译研究实验室(TRL)， 这座大楼是睡眠和呼吸神经生物学中心的所在地。生物标志物的形成 CORE将利用现有设施和现有的先进设备 翻译研究实验室。Biomarker Core将作为一个中央设施收集、处理和 分析临床项目02和03产生的大量传入血液和尿液样本 以及来自项目01和03的大鼠组织样本和分离的人类单核细胞。重要的是 Biomarker Core将确保对拟议项目收集的所有样本进行质量控制。这个 Biomarker Core的目标是：1)使不同地点之间的血液和尿液采集标准化，2)确保 样品的妥善处理和储存，3)编码、记录和监控进货样品4)确保质量 对化验的控制和5)为化验提供一个集中的设施，这些设施在以下情况下在其他设施中不可用 学院。Biomarker Core将在凯伦·特夫博士和米罗斯瓦夫博士的领导下 Mackiewicz担任共同核心领导和支持人员，由一名高级研究技术员和最低限度的 来自泰夫博士实验室的另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标记物，促炎和抗炎标记物， 黏附分子、胰岛素抵抗指数和交感神经系统活动指数。这个 BioMarker Core将进行三种类型的检测：1)高压液相色谱(HPLC) 血、尿去甲肾上腺素及去甲肾上腺素等代谢物的测定 褪黑素，一种与睡眠有关的特殊生物标志物，2)游离脂肪的分光光度分析方法 酸和甘油三酯；3)逆转录-聚合酶链式反应(RT-PCR) 组织和分离的人类单核细胞。为了避免资源重复，我们将利用其他资源 宾夕法尼亚大学的科尔斯。临床和转化性研究中心实验室常规进行酶 采用酶联免疫吸附试验(ELISA法)检测炎性细胞因子(IL-6、TNF-α)。 抗炎细胞因子(IL-10)和黏附分子(ICAM-1)。此外，胰岛素标志物 需要放射免疫分析(胰岛素、瘦素、脂联素)的抵抗力和敏感性将由 宾夕法尼亚大学糖尿病研究中心放射免疫分析核心。尿液中的异前列腺素将在 伊恩·布莱尔博士的实验室，他是化学教授和著名的氧化应激标志物专家。 血糖将在特夫博士的实验室进行分析。中央设施将确保质量控制 和成本效益，并将通过建立样本库来支持未来的项目，样本库将 可供未来调查使用。

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.