Bioanalytical Core

核心摘要 生物分析核心的目的是为人类受试者的项目#1-4提供分析能力 和小鼠败血症模型，包括分离的白细胞的基因组、生化和细胞功能分析 种群、血浆和感兴趣的组织(如骨骼肌)(S)。核心将：1)表征白细胞 群体表型；2)检测急性时相蛋白、血浆细胞因子和选择生长因子 浓度；3)获得一致的全基因组和基因特异性表达分析图谱 丰富的细胞群体和组织；4)评估肌肉中的蛋白质分解代谢和线粒体生物能量学 通过测量线粒体功能、活性氧物种和分解代谢-萎缩相关的组织 路径；以及5)为所有项目提供中央科学分析平台。 目的1.建立人和小鼠脓毒症模型的血液和组织白细胞表型。我们会 关注脓毒症患者全血和骨骼来源的早期髓系细胞群 脓毒症小鼠的骨髓和脾。 目的2.提供血浆或血清急性时相蛋白、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 Analyzer 3.1 xPONENT“系统和标准的ELISA微孔板阅读器。 目的3.测定人和小鼠全基因组和单个基因的表达 白细胞和精选的组织样本。我们将使用Affymetrix评估外显子水平的全基因组表达 基因芯片(R)技术(HH/2)和用纳米串测定选定的白细胞基因的表达 NCounter(R)。 目的4.用新的和标准的方法评估分解代谢、生物能量学和选择萎缩的途径 方法论。这些方法学将使我们能够更好地了解分子和生化变化。 与人类受试者和小鼠模型的肌肉萎缩和能量下降有关。 目标5.开发创新的新方法，为未来提供一个中心科学平台 发现号。核心将与所有项目合作，将动态科学知识和 发现并决定用于未来分析的新的创新方法。 此外，分析研究核心将集中需要复杂和 质量保证超出了单个项目的可用范围。它将保证数据的准确性和标准化 对于P50中心来说，复杂的分析方案将是经济高效的。

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.