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提供分析能力败血症的鼠模型，包括基因组，生化和细胞函数分析的分析白细胞感兴趣的种群，血浆和组织（例如骨骼肌）。核心将：1）表征白细胞人口表型； 2）测量急性期蛋白质，血浆细胞因子和选择生长因子浓度； 3）从富集的细胞群体和组织； 4）评估肌肉中蛋白质分解代谢和线粒体生物能力通过测量线粒体功能，活性氧和相关的分解代谢 - 途径； 5）为所有项目提供中央科学分析平台。目的1。来自人类和败血症鼠模型的表型血液和组织白细胞。我们将专注于早期的髓样细胞群体源自败血症的人类全血和骨骼带有败血症的小鼠骨髓和脾脏。目标2。提供等离子或血清急性期蛋白，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 Analyzer 3.1 Xponent“系统和标准ELISA微板读取器。目的3。确定人和鼠的全基因组和个体基因表达白细胞和选择组织样品。我们将评估外观级别的，全基因组的表达 Genechip（R）技术（HH/2），并通过纳米串来确定所选白细胞基因的表达 ncounter（r）。目标4。评估新颖和标准的分解代谢，生物能学和选择萎缩的途径方法论。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。开发创新的新方法并为未来提供中心科学平台发现。核心将与所有项目合作，以整合动态科学知识和发现并决定新创新的方法，以供将来分析。此外，分析研究核心将集中需要复杂的分析能力和超出各个项目的质量保证。它将确保准确性和标准化复杂的分析协议将对P50中心具有成本效益。