Core B: Laboratory Testing Core The Testing Laboratory Core B has the goal to serve every project in the Program. It will provide an array of assays assessing fibrinolysis, coagulation, anticoagulant activities, and plasma lipid profiles. The Core will also provide assays of inflammatory cytokines, and adipokines as well as with established kits for mouse proteins associated with thrombosis, atherosclerosis and obesity. The specific assays that are available for each of the projects are indicated below. Core B will also coordinate the high through-put screening of small molecules by the Center for Chemical Genomics. Project 1: Core B will perform analysis of plasma PAI-1 levels for both active and inactive PAI-1 and of plasminogen activator antigen and activity levels. In addition, plasma lipid profiles will be routinely determined and immunoassays of various inflammatory cytokines as well as adipokines will be measured in a mutiplex format using the Luminex Technology. High through-put screening of small molecule compound libraries for the development of novel PAI-1 inactivating agents will also be performed through the Center for Chemical Genomics. Project 2: Core B will perform analysis of plasma PAI-1 levels for both active and inactive PAI-1 and of plasminogen activator antigen and activity levels. Murine plasmas will also be analyzed for increased risk for thrombosis by determining the levels of plasma coagulation, and anticoagulant proteins in various strains of mice at baseline, and after venous thrombosis with or without hypercholesterolemia. D-dimer levels will also be measured. Procoagulant microparticles will also be measured as well as immunoassays of various inflammatory cytokines. Project 3: Core B will assist the investigators off Project 3 by assessing platelet function including bleeding times and platelet aggregation in mice. Fibrinolytic parameters will be determined including PAI-1 and plasminogen activator levels. Immunoassays of various inflammatory cytokines will also be measured in a mutiplex format using the Luminex Technology. Project 4: Core B will assist the investigators of Project 4 by developing novel, assays for coagulation/fibrinolysis and platelet function in Zebrafish. These assays will include immunoassays and functional assays such as clotting times as well as factor assays. The overall goals of the Testing Laboratory Core B are to enhance the success of each individual project by providing rigorous, standardized assays and to ensure that the combined Program is stronger than each of its individual parts.
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