Bioanalytical Core

核心摘要 生物分析核心旨在为人类受试者中的项目1 -4提供分析能力 和败血症的小鼠模型，包括分离的白细胞的基因组、生物化学和细胞功能分析 感兴趣的群体、血浆和组织（例如，骨骼肌）。核心将：1）表征白细胞 群体表型; 2）测量急性期蛋白，血浆细胞因子，并选择生长因子 浓度; 3）从以下获得一致的全基因组和基因特异性表达分析谱 富集的细胞群和组织; 4）评估肌肉中的蛋白质催化剂和线粒体生物能量学 组织通过测量线粒体功能，活性氧，和分解代谢萎缩相关 5）为所有项目提供一个中央科学分析平台。 目标1.对人和小鼠脓毒症模型的血液和组织白细胞进行表型分析。我们将 专注于从脓毒症患者的全血和骨中获得的早期骨髓细胞群， 脓毒症小鼠的骨髓和脾脏。 目标2.提供血浆或血清急性期蛋白、细胞因子和生长因子浓度 使用Luminex xMAP”技术和ELISA进行临床和实验室研究的测量。我们 将使用MILLIPLEX分析仪3.1 xPONENT系统和标准ELISA酶标仪。 目标3。确定人类和小鼠的全基因组和个体基因表达 白细胞和选择的组织样本。我们将评估外显子水平，基因组范围内的表达与Affyssin GeneChip（R）技术（HH/2），并通过nanoString确定选定白细胞基因的表达 nCounter（R）. 目标4。使用新的标准药物评估肌萎缩的催化剂、生物能量学和选择途径， 方法论。这些方法将使我们更好地了解分子和生物化学变化 与人类受试者和鼠模型中的肌肉萎缩和能量下降相关。 目标5。开发创新的新方法，并为未来的研究提供一个中心科学平台。 的发现核心将与所有项目合作，整合动态科学知识， 发现和决定新的创新方法用于未来的分析。 此外，分析研究核心将集中分析能力，需要复杂的， 质量保证超出了个别项目的范围。它将保证准确性和标准化， 复杂的分析协议，这将是具有成本效益的P50中心。

CORE ABSTRACT The purpose of the Bioanalytical Core is to provide analytical capability for Projects #1-4 in human subjects and murine models of sepsis, including genomic, biochemical, and cell-functional analysis of isolated leukocyte populations, plasma, and tissue(s) of interest (e.g., skeletal muscle). The Core will: 1) characterize leukocyte populations phenotypically; 2) measure acute-phase protein, plasma cytokine, and select growth factor concentrations; 3) obtain a consistent genome-wide and gene-specific expression analysis profile from enriched cell populations and tissues; 4) assess protein catabolism and mitochondrial bioenergetics in muscle tissues by measuring mitochondrial function, reactive oxygen species, and catabolism-atrophy related pathways; and 5) provide a central scientific analysis platform for all projects. Aim 1. To phenotype blood and tissue leukocytes from humans and murine models of sepsis. We will focus on early myeloid cell populations derived from whole blood in humans with sepsis, and from bone marrow and spleen in mice with sepsis. Aim 2. To provide plasma or serum acute-phase protein, cytokine, and growth factor concentration measurements for clinical and laboratory studies using Luminex xMAP" technology and ELISA. We will use a MILLIPLEX Analyzer 3.1 xPONENT" System and standard ELISA microplate readers. Aim 3. To determine genome-wide and individual gene expression in both human and murine leukocytes and select tissue samples. We will assess exon-level, genome-wide expression with Affymetrix GeneChip(R) technology (HH/2) and determine expression of selected leukocyte genes by nanoString nCounter(R). Aim 4. To assess catabolism, bioenergetics, and select pathways of atrophy with novel and standard methodologies. The methodologies will allow us to better understand molecular and biochemical changes related to muscle atrophy and energy decline in human subjects and murine models. Aim 5. To develop innovative new methodologies and provide a central scientific platform for future discovery. The Core will collaborate with all projects for the integration of dynamic scientific knowledge and discovery and decide on newly innovative methodologies for future analysis. Additionally, the Analytical Studies Core will centralize the analytical capabilities that require sophistication and quality assurance beyond that available to individual projects. It will assure accuracy and standardization of complex analytical protocols that will be cost-efficient for the P50 Center.