Practicum Three, August 2000. Novel applications of carbon-13 indirect detection NMR to metabolic tracer studies in small tissue samples. Practicum Two, December 1998. Proton and platinum-195 NMR spectroscopic characterization of the products formed in re

Practicum three. Carbon-13 tracers are widely used in the study of intermediary metabolism. Thus 13C NMR "isotopomer analysis" has become important for investigating the Tricarboxylic acid (TCA) cycle and it's many intersecting pathways. The primary weakness of this technique is the relative insensitivity of 13C NMR. As a result, experiments on genetically altered mice, cell cultures and clinically relevant human samples have garnered only limited attention because of the impractical instrument time that is required for such small samples. Proton observation of 13C is a more sensitive approach than 13C direct observation. Although this technique has been applied to the structural analysis of proteins for nearly 2 decades, it has only recently been use to answer metabolic questions.; The first chapter of this report deals with the use of J-resolved heteronuclear single quantum coherence (J-HSQC) spectra for isotopomer analysis. This 2D indirect detection experiment gave the same results as 13C direct detection, but in less time. The second chapter reports on the use of indirect detection for the measurement of fluxes through the TCA cycle of the isolated perfused mouse heart. We observed ample sensitivity to measure TCA cycle flux and glutamate exchange flux in the mouse heart. Kinetic data was collected with a temporal resolution of 22 s compared to 3 min typically required for a 13C NMR experiment on a perfused rat heart. Our data was compared to similar data from larger rodents and indicates a higher TCA cycle flux for mice.; Practicum two. Three peptide derivatives of ethylenediamine were synthesized and reacted with K2PtCl4. Proton, 13C, and 195Pt NMR were used to determine the solution structures of the resulting mixture of compounds. The platinum binding characteristics over several pH(s) and temperatures were also reported.; Practicum one. A novel polyamide containing the macrocycle DO2A in its backbone was synthesized as a potential angiographic contrast agent. The polymer and a monomeric model compound were structurally characterized by NMR and mass spectroscopy. The molecular weight of the polymer was determined by size exclusion chromatography. Potentiometry and O17 NMR were used to determine the Gd3+ binding properties of model compound.