The Effect Of Plasma Metabolome Following Dietary Depletion And Supplementation Of Thiamine Pyrophosphate

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Exam 2 Guoqing Wang 1. Metabolomic analysis of changes to plasma metabolome following dietary depletion and supplementation of thiamine pyrophosphate (TPP) in mice Experimental design Male C57BL/6J mice (No. 11401300006402), weighing 20 ± 2 g, will be used for this experiment. 7-month old mice will be maintained in a room at 23 ± 2°C and a relative humidity of 50 ± 10%, with a natural light-dark cycle. Food (6 mg thiamine-HCl as the form of TPP) and water will be provided ad libitum. After acclimatization for 1 week, the mice will be divided into two groups (n = 10 in each group) based on dietary difference: the dietary TPP depletion group (no TPP added) and the dietary TPP supplementation group (6 mg thiamine-HCl/kg diet as the form of TPP). The supplementation level will be based on Nutrient Requirements of Laboratory Animals: Fourth Revised Edition, 1995 [1]. Plasma samples will be collected from mice in each group before feeding the experimental diets and after 10-day feeding period, the length of feeding experimental diets will be based on a previous publication [2]. Approach Plasma samples will be analyzed by H-NMR with or without filtration and by targeted quantitative by mass spectrometry (MS) according to Gregory III et al., 2013 [3]. NMR spectral will feature selected metabolites including acetyl-CoA, succinyl-CoA, acetoacetate and ribose. Expected results Compared to dietary thiamine supplementation at the recommended level, thiamine depletion will decrease

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