Opredelitev sistema bilirubin, albumin in fibroblasti V-79 z elektronsko paramagnetno resonanco : doktorska disertacija
Bratanič, Borut, 1947-
Introduction. A high unconjugated bilirubin concentration (BR) in serum could present a risk factor for permanent brain damage especially in preterm and sick term new-borns. The toxic effects of BR, ...because of very low water solubility of this at physiological pH internally H-bonded molecule, depend onits unconjugated, from albumin (ALB) displaced Ćfree BRĆ fraction. ĆFreeĆ BRis distributed among numerous compartments (cells and their membranes) and also penetrates through blood brain barrier into central nervous system of new-borns. The exploration of dynamic interactions between BR, ALB and membranes is essential for understanding of BR toxic effects on cells in living organisms. Therefore the research started with EPR (electronic paramagnetic resonance) spectroscopy study of the effects of simultaneous binding of BR and spin labelled palmitic acid on ALB. The toxic effects of BR on the fibroblast cell line (V-79) were studied by the survival and growth rate of cellular cultures. On the cellular membranes of V-79 and mouse neuroblastoma cells (Neuro 2a) the structural and functional changes were determined by EPR spectroscopy. The influence of BR on reduction processes of nitroxide radicals by V-79 cells was studied as a model of BR participation inred-ox processes. Methods. The EPR spectroscopy on Bruker 300 X-band (9 GHz)instrument in 100 mW microwave field, at 100 kH frequency and 0.1 mT amplitude of modulation, with the spin labelling of ALB and cellular membraneswas applied. With the binding of BR (Merck, art 24520) and the spin probe (2-(3-carboxypropil)-2-undecil-4,4- dimetiloxazolidin-3-oxyl palmitic acid (HFASL (10,3)) on ALB (bovine albumin, Sigma, No. A-9647) in PBS water solution (pH 7.4č osmolality 290 mmolžkg), conformational changes on ALB molecules were detected by changes in hyperfme splitting of albumin bound spinlabel spectra. (Abstract truncated at 2000 characters.)
Type of material - dissertation
; adult, serious
Publication and manufacture - Ljubljana : [B. Bratanič], 1999
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