Bioanalytical Core

生物分析核心

基本信息

批准号：
8740716
负责人：
CHRISTIAAN LEEUWENBURGH
金额：
$ 28.11万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-01 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8740716
关键词：
3-methylhistidine Actins Acute-Phase Proteins Affect Albumins Angiopoietin-1 Apoptosis Atrophic Basic Science Biochemical Bioenergetics Biological Biological Assay Blood Bone Marrow Brain natriuretic peptide C-reactive protein CCL2 gene CD14 gene CXCL12 gene CXCR3 gene Calpain Caspase Catabolism Cells Chronic Clinical Research Clinical Sciences Complex Concentration measurement Critical Care Critical Illness Custom DNA copy number Data Quality Ensure Enzyme-Linked Immunosorbent Assay Equipment Exons Flow Cytometry Future Gene Chips Gene Expression Gene Expression Profiling Genes Genomics Growth Factor HLA-DR Antigens HMGB1 gene Human Human Identifications IL4R gene IL8 gene ITGAM gene Immunoassay Immunologic Markers Immunosuppression In Situ Individual Inflammation Interferons Interleukin-1 Interleukin-10 Interleukin-6 Intervention Trial Knowledge Discovery Laboratories Laboratory Study Leukocytes Longitudinal Studies Measurement Measures Methodology Mitochondria Mitochondrial DNA Modeling Molecular Morphology Mus Muscle Muscular Atrophy Myelogenous Myeloid Cells Myosin ATPase NOS2A gene Operative Surgical Procedures Paralysed Pathway interactions Phenotype Plasma Population Production Proteins Proteolysis Protocols documentation Reactive Oxygen Species Reader Research Research Personnel Resolution Role Sepsis Serum Services Skeletal Muscle Spleen Standardization Syndrome System TNF gene TNFRSF5 gene Techniques Technology TimeLine Tissue Sample Tissues Translational Research Vascular Endothelial Growth Factors Western Blotting Whole Blood clinical application cost cytochrome c oxidase cytokine genome-wide human subject innovation interest interleukin-12 subunit p40 monocyte multicatalytic endopeptidase complex nano-string novel polypeptide C procalcitonin progenitor quality assurance urinary

项目摘要

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.

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提供分析能力和脓毒症小鼠模型，包括分离白细胞的基因组、生化和细胞功能分析群体、血浆和感兴趣的组织（例如骨骼肌）。核心将：1）表征白细胞群体表型； 2) 测定急性时相蛋白、血浆细胞因子、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谱丰富的细胞群和组织； 4) 评估肌肉中的蛋白质分解代谢和线粒体生物能学通过测量线粒体功能、活性氧和分解代谢萎缩相关的组织途径； 5) 为所有项目提供中央科学分析平台。目标 1. 对人类和小鼠脓毒症模型的血液和组织白细胞进行表型分析。我们将专注于源自脓毒症患者全血和骨骼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并选择组织样本。我们将使用 Affymetrix 评估外显子水平、全基因组表达 GeneChip(R) 技术 (HH/2) 并通过 nanoString 确定选定白细胞基因的表达 n计数器(R)。目标 4. 评估分解代谢、生物能量学并选择新颖和标准的萎缩途径方法论。这些方法将使我们能够更好地理解分子和生化变化与人类受试者和小鼠模型中的肌肉萎缩和能量下降有关。目标 5. 开发创新的新方法并为未来提供中央科学平台发现。该核心将与所有项目合作，整合动态科学知识和发现并决定用于未来分析的新创新方法。此外，分析研究核心将集中需要复杂性和准确性的分析能力。质量保证超出了单个项目的质量保证。它将确保数据的准确性和标准化复杂的分析方案对于 P50 中心来说具有成本效益。