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The rat ovarian perfusion model with bursa removed and intact was used to further characterize the effects of interleukin-1 beta (IL-1 beta) and the natural IL-1 receptor anatagonist (IRAP) on ovulation, steroidogenesis, and prostaglandin production. Twenty-six- to 27-day-old female Sprague-Dawley rats were injected sc with 25 IU PMSG, and 48 h later, the right ovary was dissected (with bursa removed and intact for various experiments) and placed in the perfusion chamber. Ovaries were exposed to various doses of IL-1 beta alone, IL-1 beta with LH, and IL-1 beta with LH and isobutylmethylxanthine (IBMX). The natural IL-1 receptor antagonist was also added to the chambers with LH and IBMX. IL-1 beta at 0.8 (n = 3) and 8.0 (n = 4) nM did not induce LH-independent ovulation in PMSG-stimulated ovaries with bursa removed. In bursa-intact perfusions (n = 3), one ovulation was produced in each compared to control ovaries with bursa intact (n = 3) given an ovulatory trigger of LH alone [2.3 +/- 0.6 (+/- SD) ovulations; P < 0.02]. IL-1 beta enhanced, in a dose- and gonadotropin-dependent fashion, the production of prostaglandin E2 (PGE2) in PMSG-stimulated ovaries with bursa removed given an ovulatory trigger of LH and IBMX compared to that in controls. PGF2 alpha and 6-keto-PGF1 alpha were also modulated by IL-1 beta. Estradiol and progesterone production were not affected. The natural IRAP inhibited ovulation (7.8 +/- 3.9 ovulations vs. 12.4 +/- 1.5; P < 0.04) in PMSG-stimulated ovaries given LH and IBMX as the ovulatory trigger compared to that in controls. This inhibition of ovulation was not associated with reduced steroid or PG levels. IL-1 beta appears to play a potentially significant role in the process of ovulation. The functional importance of the bursa in this model is highlighted in this study. IL-1 beta modulates PG, but not steroid, production. IRAP inhibited ovulation without significantly affecting PG or steroid production.

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Data from five years of radioimmunoassay (RIA) screens and gas chromatographic/mass spectrometric (GC/MS) quantitation of blood, or blood and matched (i.e. concurrently collected) urine specimens for cannabinoids have been used for two distinct evaluations. The first part, which is the subject of this study, considers comparative results from RIA and GC/MS analysis of the specimens. RIA analysis (Abuscreen urine cannabinoids kit) of methanol extracts of blood with extracted blood calibrators (122 specimens) was compared to analysis of nonextracted blood with urine calibrators (108 specimens). RIA of methanol extracts of blood was a reliable assay (limit of detection = 17 ng/mL; interassay coefficients of variation of 2-10%). RIA of nonextracted GC/MS negative blood gave significant displacement of labeled antigen, which precluded accurate quantitation using the urine-based calibrators. Comparison of RIA cannabinoid concentrations to 11-nor-delta 9-tetrahydrocannabinol-9-carboxylic acid (COOH-THC) concentrations measured by GC/MS yielded correlation coefficients of 0.68 and 0.23 for the methanol extract and nonextract procedures, respectively. Comparison of the qualitative accuracy of the two techniques, however, resulted in similar discrimination of GC/MS positive and negative specimens when the 20-ng/mL extracted blood calibrator was used as the cutoff for methanol-extracted blood specimens, and the 100-ng/mL urine calibrator was used as the cutoff for nonextracted blood specimens. Although qualitatively similar, the methanol extract procedure resulted in quantitatively superior analysis.