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The aim of this study was to investigate the relationships between follicular nitric oxide (NO) metabolite concentrations and several related variables, with special reference to follicular interleukin-1beta (IL-1beta). The follicular fluid from the leading and secondary follicles was collected individually from 20 women undergoing in-vitro fertilization (IVF) treatment, and the concentrations of nitrite (NO2-) and nitrate (NO3-) were determined fluorometrically using 2,3-diaminonaphthalene. Both follicular nitrite (r = 0.42, P < 0.01) and nitrate (r = 0.49, P < 0.001) were found to be significantly correlated with follicular IL-1beta concentrations. There were also significant positive correlations between follicular nitrate and the number of oocytes retrieved (P < 0.01) and serum oestradiol concentration on the day of human chorionic gonadotrophin (HCG) administration (P < 0.05). When follicular cells were incubated in vitro with 10 ng/ml of IL-1beta for 24 h, nitrate generation was significantly (P < 0.01) elevated compared with the control. In conclusion, our study demonstrates that follicular IL-1beta and the number of developing follicles are significant variables that affect follicular NO concentrations, and points to the possible contribution of IL-1beta to NO generation in human preovulatory follicles.

The present study was to clarify the relationship between voluntary exercise and follicular growth or ovulation. Rats kept for 4 weeks in a rotating drum with free access to the wheel, food and water ran 2-12 km per day. The number of ova shed after superovulation treatments and the number of non-atretic follicles were not influenced by voluntary exercise. These experiments demonstrate that spontaneous voluntary exercise does not affect either the number of ova shed or the number of non-atretic follicles in superovulating rats or control rats.

The effects of body weight and population density on the number of ova shed were examined in the superovulation-treated adult rat in order to clarify the variation of the number of ova shed. Twelve-week-old female rats, weighing 182-277 g, were injected with 40 i.u. of PMSG on metestrus, with sodium pentobarbital on diestrus ("critical period"), and with 40 i.u. of hCG, on estrus. Twenty hours after the injection, the number of ova shed in an ampulla was observed. There was no correlation between body weight and the number of ova shed (r = 0.004). The rats were kept in at three, six or nine rats per cage (260 X 380 X 180 mm) from weaning at 30 days old to 12 weeks, with superovulation treatment. There was no significant difference in the number of ova shed among the various population densities.

A new treatment for inducing superovulation in adult rats was examined. Virgin Wistar-Imamichi rats at 11 weeks of age were pretreated with PMSG on the metestrus day. Then, the animals were anesthetized with sodium pentobarbital on the expected estrus day. The maximum number of ova shed in the animals thus treated was 100 with a combination of 40 I. U. of PMSG and hCG. If the treatment with sodium pentobarbital was omitted, the maximum number remained at 43 with different combinations of PMSG and hCG dosages.