Cessation of transition-phase follicle growth in the guinea pig by follicle-regulatory protein.

The granulosa cell produces a protein inhibitor of aromatase activity (follicle-regulatory protein: FRP), which recently was purified to homogeneity. To determine the possible involvement of FRP in follicular maturation, we examined the size distribution of follicles and their morphological patterns as well as serum steroid levels after the systemic administration of FRP and/or gonadotropin to guinea pigs, which have 5-6 days between luteolysis and ovulation in a 16-day cycle. FRP was partially purified from porcine follicular fluid by ammonium sulfate precipitation (0-35%), Dye Matrex Orange A Chromatography, dialysis, and lyophylization. To investigate the effect of pregnant mare's serum (PMS) during the periovulatory period in follicular development, adult guinea pigs underwent unilateral ovariectomy on Days 10, 12, and 14 of the estrous cycle (N = 6 each). Guinea pigs were injected twice daily with vehicle or PMS (5 IU) and 2 days thereafter the remaining ovaries were removed. Another group of guinea pigs received, in addition, intraperitoneal injections of FRP (1 mg) each morning from Day 8 of estrus until they were killed. The resected ovaries were fixed, embedded in paraffin, serially sectioned (7 micron), and stained with Azan for comparative study via light microscopy. All follicles greater than 400 micron were classified by size, and the atretic pattern was determined by mural granulosa cell pyknosis and antral sloughing. The distribution of follicular size was not affected by hemicastration at Day 10, although the percentage of total atretic follicles decreased. In the PMS-treated group, there was a significant decrease in the number of viable follicles (700-899 micron) after hemicastration. Also pronounced in follicles of this size was the lack of mid-atretic follicles. After injections of FRP for 3 or 5 days, the overall number of follicles was almost doubled as compared to the number found in the normal ovary. Additionally, there was a significant increase in the percentage of follicles that were recently atretic, although the total percentage of atretic follicles was unchanged. After hemicastration at Day 10 followed by FRP treatment for 2 days, the total percentage of atretic follicles in the remaining ovary decreased to 18% compared with 35% in the normal ovary, 46% in the hemicastrated plus PMS-treated group, and 38% in the hemicastrated and PMS- and FRP-treated group (all p less than 0.01). Treating the hemicastrated animal with PMS increased the percentage of atretic follicles in all groups.(ABSTRACT TRUNCATED AT 400 WORDS)

Inhibition of FSH augmented adenylate cyclase activity in porcine granulosa cells by ovarian protein.

We sought to evaluate the effects of a fraction of porcine follicular fluid, termed follicle regulatory protein (FRP), on FSH-induced adenylate cyclase activity in porcine granulosa cell membranes using Gpp(NH)p and forskolin as pharmacological probes of adenylate cyclase activity. Without FSH treatment, the addition of 100 micrograms/ml of the FRP fraction induced a significant decrease in Gpp(NH)p stimulated cyclase activity while maximal inhibition of cAMP formation was achieved with 1 mg/ml of FRP. Granulosa cells cultured with FSH reached a maximum in adenylate cyclase activity at 20 min which returned to baseline by 45 minutes. FRP induced a reduction in adenylate cyclase activity during this same interval of time. Adenylate cyclase activity of cells treated with FRP was unchanged in the presence of methyl-isobutal-xanthine. Further, when FRP was heated (56 degrees C, 45 min) or precipitated with 10% TCA, it was unable to inhibit adenylate cyclase. The 50% inhibitory dose (ID50) for FRP inhibition of adenylate cyclase activity in cells stimulated with Gpp(NH)p was 80 micrograms/ml and 500 micrograms/ml when granulosa cells were preincubated with FSH prior to Gpp(NH)p stimulation. The ID50 for the FRP inhibition of forskolin stimulated adenylate cyclase activity was 500 micrograms/ml. Gpp(NH)p stimulated adenylate cyclase activity was more sensitive than forskolin stimulated activity to inhibition by FRP. In conclusion, the data presented here demonstrate that a partially purified fraction of porcine follicular fluid inhibited FSH responsive adenylate cyclase activity in porcine granulosa cells.

Double-blind comparison of meclofenamate sodium with codeine and placebo for the pain of episiotomy.

Meclofenamate sodium, a nonsteroidal anti-inflammatory agent, was compared at two dose levels (100 mg and 200 mg) with codeine (60 mg) and placebo in a double-blind, randomized study of 218 women after normal vaginal delivery. The purpose was to determine the analgesic efficacy and safety of meclofenamate sodium for the short-term treatment of acute episiotomy pain. Meclofenamate sodium was significantly better than placebo in most measures of pain relief and reduction of pain intensity. The 100-mg dose of meclofenamate sodium was significantly better than codeine in relieving pain. Adverse experiences with the study medications were minimal (6.4%). Patients receiving codeine reported more side effects than did those receiving either dose of meclofenamate sodium. Meclofenamate sodium is a safe, effective analgesic for acute episiotomy pain.