As with the first three years of this MBRS grant, the overall objectives of this program is to stimulate minority undergraduate students to become biomedical research scientists who are motivated to pursue further study in biomedical and health-related graduate schools and to enter competitive biomedical research careers. To accomplish this goal, we will continue to expose and directly involve minority students in on- going biomedically related research projects described in the proposal. The specific scientific aim of this research project is to measure the extent to accumulation of defective amino acids (D-aspartate, D- glutamate, L-isoaspartate) in aged and dysfunctional proteins and in proteins of heat-processed foods. Various types of proteins are being examined, namely: cerebral spinal fluid and brain proteins from victims of neurological disorders such as Alzheimer's Disease, cartilage from osteoarthritic human knees, and milk proteins from conventionally heated vs microwaved milks. A primary focus of this research is Alzheimer's disease. Our preliminary findings of differences in the D-aspartate content of cerebral spinal fluids (CSF) of Alzheimer's versus normals could have significant consequences for possible early diagnosis of the disease. Presently there is no definitive diagnosis for Alzheimer's disease until the patients has died and a neuropathological examination of the brain is done. If we confirm our preliminary findings, then analysis for D-Asp in lumbar CSF spinal taps of suspected Alzheimer victims could possibly be a highly significant break-through as a diagnosis for the disease. In addition, we are also investigating mechanisms for racemization (change in configuration from L-to D- or vice versa) of aspartate and possible ways of reversing D-Asp back to normal L-Asp using in damaged proteins (e.g. myelin basic protein) back to the normal L-Asp. Further confirmation of this will have significant consequences for possible mechanisms of treatment and repair of damaged proteins. Measurement of the D-amino acids is accomplished by chromatographic techniques such as gas chromatography (GC), high performance liquid chromatography (HPLC), and capillary electrophoresis (CE), or by enzymatic techniques employing spectrofluorometric analyses. L- isoaspartate is measured by an enzymatic assay procedure using protein carboxyl methyltransferase (PCM).
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