The overall goal of this research Program is to develop a deeper understanding of the mechanisms linking oxidation and inflammation to cardiovascular disease. The Project Leaders in this Program have contributed significantly to an enhanced understanding of specific enzymatic participants in nitric oxide (NO) production and oxidant formation during inflammation. In an effort to expand the scope and clinical relevance of these individual efforts, we have built an integrated research Program that explores the interactions between oxidant-generating pathways, their biological consequences, and overall mechanisms linking inflammation and oxidation to the pathogenesis of cardiovascular diseases. Our Program Project, which is comprised of 4 interrelated projects and 4 cores, is predicated upon the hypothesis that oxidative stress is mechanistically linked to the development of cardiovascular disease. To test this hypothesis, Project 1 employs a combination of genetic, biophysical and clinical studies to define the potential role of myeloperoxidase and specific oxidation pathways in development of cardiovascular disease in humans. Project 2 is thematically linked to Project 1, and explores structural determinants for pro-oxidant and pro-inflammatory activities in the copper-containing protein ceruloplasmin, and their relevance to the atherosclerosis phenotype in humans. The theme of inflammation and oxidation continues in Project 3, which will study mechanisms through which distinct leukocyte-generated oxidants modulate activity of plasminogen activator inhibitor-1, a protease inhibitor that participates in responses to oxidative tissue injury. In Project 4, the functional significance of polymorphisms occurring in endothelial and inducible NO synthases will be explored, along with the potential therapeutic utility of engineered NO synthases in models of carotid artery restenosis. A unique and unifying thread throughout the Program is the in-depth examination of shared clinical samples and connecting database from patients undergoing cardiac catheterization and long-term follow-up within our institution. Three scientific cores (Mass Spectrometry; Protein Engineering/Biophysics; Animal Model) and an Administrative/Biostatistics Core further strengthen the research Program by providing multiproject support, expertise and service in a cost-effective manner. The proposed Program Project will yield a greater understanding of the role of oxidation in normal physiologic processes and disease pathogenesis. It may also lead to important insights for atherosclerosis risk assessment, diagnosis and therapy.