A key objective of the current funding of the ROl grant is to advance our understanding of the regulation of muscle mitochondrial biogenesis and its role in type 2 diabetes. The proposal also addresses the gap in our understanding on regulation of synthesis of individual proteins in muscle mitochondria and plasma. The original proposal received a top score (priority score 112, 0.2 percentile) of NIH review panel. A unique measurement that is only performed in our laboratory is measurement of synthesis of individual muscle mitochondrial and plasma proteins. Although most of the human studies as outlined in our original proposal are completed, the purification of individual proteins and their isotope measurements are not progressing at sufficiently fast rate. The main problems are related to a very old LC system and not having sufficient numbers of isoelectric focusing systems. Purchasing a new LC system to replace the current outmoded system and an additional isoelectric focusing system will enable us to complete the analysis of samples in a timely manner. The important questions that will be answered by the above analysis are: 1) the role of amino acids and insulin regulating translational rates of mitochondrial proteins encoded by nuclear and mitochondrial genes and 2) whether amino acids and insulin have any specific effect in regulating plasma protein synthesis. Studies performed in our laboratory and that of others have clearly shown that greater understanding of the regulation of mitochondrial biogenesis is critical to understand the pathophysiology of the many metabolic diseases. The results from the proposed studies will substantially advance our understanding on mitochondrial biogenesis in humans. It is well known that many plasma proteins are implicated as potential cardiometabolic risk factors. Completion of the analysis of the samples from the completed human studies will offer insight into the regulation of key plasma proteins. Proteins are the functional molecules and gene products. The current grant is addressing a huge gap in our understanding about proteins by studying individual proteins with variable functions. The proposed equipment purchase will replace outmoded equipment and enhance our ability to complete analysis of samples from human studies in a timely manner.