Bioanalytical Core

生物分析核心

• 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
• 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

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.

核心抽象