Analytical and Morphological Core

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The primary purpose of the Analytical and Morphological Core (Core B) is to provide established and centralized assays and expert personnel for projects of the Program Project Grant (PPG), which includes analytical chemistry, enzyme-linked immunoassay (ELISA), radioimmunoassay (RIA), real-time polymerase chain reaction (RT-PCR), electrophoretic mobility shift assay (EMSA) and histological, morphological and imaging analysis. Analytical chemistry covers measurement of nitrites/nitrates, creatinine, hydroxyproline and kallikrein activity. ELISA will be used to measure N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP), prostanoids, cytokines, unnary protein, etc. RIA will be used to measure plasma renin activity and renin and angiotensin concentrations, cyclic adenosine (cAMP) and guanosine monophosphates (cGMP). RT-PCR and EMSA will be performed to measure gene expression and nuclear factor kappaB ( N F K B ) activation and translocation. All chemical analyses and bioassays will be performed on samples obtained from tissues, body fluids or cultured cells. Histomorphology includes tissue processing, embedding, cutting, histological and immunohistochemical staining and photography for measurement of a) infarct size, cardiomyocyte size, capillary density, interstitial and perivascular collagen; 2) inflammatory cell infiltration, adhesion molecules, markers of oxidative stress, apoptosis and necrosis in the vasculature, heart and kidney; and c) periodic acid Schiff (PAS) staining for kidney lesions. The Project Leaders will analyze the histological data in a blinded fashion with the assistance of Core B. In addition, the Analytical and Morphological Core will relieve Project Leaders from performing these labor-intensive and time-consuming analyses themselves and instead allow them more time to pursue their scientific efforts. The anticipated Analytical and Morphological Core's effort for each project is estimated as Project I, 55%; Project 11, 30%, Project HI, 10%; and Project IV, 5%. RELEVANCE (See instructions): By centralizing analytical and morphological studies for Project Leaders, Core B will help integrate participating projects by providing consistent, reliable and objectively analytic and morphological assays, which will promote both quality and productivity of participating projects. Core B will ensure cost efficiency in terms of labor and laboratory facilities/equipment to avoid duplication of efforts by individual Project Leaders.