The main function of the Core is to provide state-of-the-art analytic methods to advance the proposed research in all three projects of the Program Project. We will provide reliable measurements and analyses of blood glucoregulatory, appetitive, and circadian hormones and other metabolite from human subjects who are affected by aging and/or sleep duration and quality in Projects 1 and 2. Assays will include leptin, ghrelin, gastric inhibitory polypeptide (GIP), C-peptide, cortisol, melatonin (from blood and saliva), 6-sulfatoxymelatonin (MTS6 in urine), glucose, insulin, lactate, pyruvate, free fatty acids (FFA), triglycerides, LDL, HDL, HbA1c, and NAD+. We will all also provide metabolic phenotyping methods to study tissue-specific and cellular metabolic flux in both the animal tissues and human blood samples and biopsies. The analytical procedures include high- performance liquid chromatography and mass spectrometry for metabolite analyses, cellular bioenergetics, real time monitoring of cellular redox state, biochemical analysis of sirtuin activity, gene expression analysis using quantitative PCR methods, as well as transcriptome analyses using next-generation sequencing for animal models in Project 3. All analytical methods have been validated and operational in our laboratory. The Metabolic and Molecular Core will provide critical support for the implementation of all Projects, explore potential use of novel analytical techniques, and will be closely integrated with the Analysis Core to ensure timely and safe archival of laboratory data.
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