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In vitro metabolism of ovarian steroids by bovine whole blood and plasma
Authors:RR Lesniewski  JD Kirsch  LE Mrozinski  DH Hellwig  J Kotwica  GL Williams
Institution:Departments of Animal Science and Biochemistry North Dakota State University Fargo, North Dakota 58105 USA
Abstract:Experiments were conducted to evaluate the effects of time and temperature on the potential of bovine whole blood (WB) or plasma (PL) to metabolize the ovarian steroids progesterone, estradiol-17β and testosterone. During a radioimmunoassay study (Experiment 1), we observed a temperature and time-dependent reduction (P<0.001) of plasma progesterone concentrations in samples incubated as WB at 5, 15, 25, or 35C for up to 48 hr. Most notable was the observation that 27% of progesterone present in controls was lost when WB was incubated at 5C for 48 hr and a 17% reduction was observed when PL samples were incubated at 35C for 48 hr. Immunoreactive estradiol-17β concentrations (Experiment 2) in PL and WB incubates were not affected by time or temperature. However, immunoreactive testosterone concentrations increased more than 3-fold by 48 hr in WB incubates held at 35C. To examine the latter observation further, 3H-progesteone was incubated with WB at 35C, followed by extraction and thin-layer chromatography (Experiment 3). Results generally supported RIA findings and revealed the presence of significant 17α-hydroxylase, 17–20 lyase and aromatase activity. Heretofore this has not been considered to occur outside major steroid metabolizing organs.
Keywords:Reprint requests: G  L  Williams  Texas A&M University Agricultural Research Station  HCR-2 Box 43C  Beeville  Texas 78102
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