Donald A. Shutt

Assessment of possible luteolytic effect of intra-ovarian injection of prostaglandin F2α in the human

A group of five patients awaiting laparoscopic tubal diathermy were followed by daily assay of luteinising hormone (LH) and progesterone. Between five and eight days after the LH peak, prostaglandin F-2ALPHA (PGF-2ALPHA) was injected into either the corpus luteum or the ovarian stroma. Doses of 100 mu-g into the corpus tuteum, 1000 mu-g into the adjacent stroma and 500 mu-g into an indeterminate ovarian structure had no effect on peripheral plasma progesterone levels or uterine bleeding. An injection of 500 mu-g or 1000 mu-g given unequivocally into the corpus luteum produced a rapid and profound fall in plasma progesterone levels, the nadir coinciding with the onset of uterine bleeding which commenced 24 hours after the injection and persisted for more than seven days. Injection of 100 mu-g in the same volume of saline had no such effect. Despite continued bleeding plasma progesterone levels returned to normal luteal levels for three days and then fell again.

Radioimmunoassay of progesterone, 17-hydroxyprogesterone, estradiol-17β and prostaglandin F in human corpus luteum

Radioimmunoassay procedures have been adapted for the assay of progesterone, 17-hydroxyprogesterone, estradiol-17beta, and prostaglandin F in human corpus luteum. The method utilises a single homogenisation and extraction of the tissue followed by fractionation of the steroids on alumina, and separation of the prostaglandins of the F series from the E and A series on silica gel, prior to radioimmunoassay. An attempt has been made to validate the method for the progestins by comparison with results after fractionation of the progestins on Sephadex LH-20, for estradiol-17beta by comparison with values obtained with competitive protein-binding, and for prostaglandin F by comparison with values after additional purification. The results showed that peak concentrations of the three steroids in corpora lutea from women during the luteal phase of the menstrual cycle were comparable to those found in corpora lutea from women in early pregnancy. However, in six out of fourteen corpora lutea from non-pregnant women, prostaglandin F levels were higher than those found in corpora lutea from seven women in early pregnancy, i.e. 13-46 ng/g compared with 1-7 ng/g. Of the above six corpora lutea, four were on days 23-25 of the cycle, at a time when luteolysis would be commencing. The results in this paper support the conclusion that the corpus luteum is a major site of synthesis of the three steroids examined, although the site of synthesis of prostaglandin F is still equivocal.

The effect of intra-uterine prostaglandin F2α on corpus luteum function in the human

Prostaglandin F2α (PGF2α) was administered via a Foley catheter over a 12 hour period to 8 healthy volunteers awaiting laparoscopic sterilisation. The amount of PGF2α infused varied between 500 μg and 2000 μg every 2 hours for 6 doses. Plasma progestins and oestradiol 17β, and urinary estrogens and pregnanediol were assayed throughout the study period. There was no evidence of luteolysis in any patient although vaginal bleeding of varying duration occurred in all women within 36 hours of administration of PGF2α.

FOETAL PLASMA STEROID CONCENTRATIONS RELATED TO GESTATIONAL AGE AND METHOD OF DELIVERY

Abstract Levels of total corticosteroids and unconjugated oestrone and oestradiol-17β in human umbilical arterial and venous plasma at the time of delivery were measured, using competitive protein-binding methods. Increased corticosteroid levels were associated with the establishment of labour and, following delivery at term (38–40 weeks), corticosteroid levels were least when the mother was not in labour and greatest when the mother was in established spontaneous labour at the time of delivery. A surge and peak in corticosteroid levels was observed at 35–37 weeks of gestation in infants delivered by elective Caesarean section and per vaginam following spontaneous labour and thus was not related to labour. Oestrone and oestradiol-17β levels in umbilical venous plasma have also been shown to change with the establishment of labour, oestrone levels increasing and oestradiol-17β levels falling, so that oestrone predominates. Following spontaneous labour with vaginal delivery, there was a significant effect of parity upon oestrogen levels, higher levels of both oestrone and oestradiol-17β occurring in infants from primiparous mothers. This effect of parity was not observed following Caesarean section, either when in early labour or not in labour. It is concluded that fluctuations in corticosteroid and oestrogen levels in the feto-placental circulation are associated with the establishment of labour and may therefore have a regulatory role in human parturition.

Utilization of a single antiserum for the direct radioimmunoassay of prostaglandins E and F in semen and prostaglandin F in amniotic fluid

Abstract Antibodies to both prostaglandin F (PGF) and prostaglandin E (PGE) were raised in rabbits after they were immunized with prostaglandin F2a conjugated to bovine serum albumin (PGF2a - BSA). The antisera were group specific although the antibodies to the F group of prostaglandins showed greater specificity than those to the E group. The antisera were sufficiently specific however to allow the direct radioimmunoassay of PGF and PGE in human semen and PGF in amniotic fluid during induced abortion. Specificity of the direct radioimmunoassay was checked by chromatographic separation of the prostaglandins prior to analysis. Estimation of the prostaglandins in the semen of 30 men attending the infertility clinic showed that 19 of the men had normal semen levels of PGE and PGF of 68± 7(SE) and 6.0 ±0.6 μg/ml respectively, as compared with data on normal fertile males, whilst the other 11 men had lower levels of 16 ±2 (SE) and 0.8±0.1 μg/ml respectively. Application of the method to amniotic fluid showed that the PGF concentration in amniotic fluid during the induction of abortion with extra-ovular saline increased from less than 0.6 ng/ml to 6.4 ng/ml when the induction-abortion intervals ranged from 6–48 hours.

The Effect of Intra-amniotic Injection of Triiodothyronine on Pulmonary Maturity in Lambs at 130 Days Gestation

Summary: Saline or triiodothyronine (T3) (50-1000 μg) was injected into the amniotic sac of 17 pregnant ewes under general anesthesia at 130 days gestation. Forty-eight h later, the lambs were delivered by hysterotomy. Cord plasma T3 and cortisol and amniotic fluid T3 were assayed, and the maturity of the fetal lung was assessed in terms of its pressure-volume response and its surfactant (lamellar body phospholipid) content. With the highest dose of T3, cord plasma T3 and cortisol were raised, and lung maturity was enhanced compared with saline-treated controls; the pressure-volume curve showed increased hysteresis on inflation and deflation, and the lung retained air on return to zero pressure. There was also an apparent, but not statistically significant, increase in the lamellar body phospholipid content of the lung. Irrespective of treatment, lungs which were more mature, in terms of their pressure-volume characteristics, tended to contain a higher proportion of lamellar body phospholipid relative to total phospholipid.

Changes in estrogen levels in maternal venous plasma after intra-amniotic infusion of Prostaglandin F2α for therapeutic abortion

Peripheral plasma levels of estrone, estradiol-17beta and estriol were measured by the method of Shutt and Cox in 10 women following intra-amniotic infusion of prostaglandin F2alpha (PGF2a) for therapeutic abortion. Initial dose was 30 mg, followed if necessary, by doses of 15 mg at 24 hours and 42 hours. Gestational age of pregnancies ranged from 14 to 19 weeks, with a mean of 16 weeks. All 10 women completely aborted. Mean induction-abortion interval was 24 + or - 12 hours. The mean estrone, estradiol 17beta and estriol levels declined to about half of the pre-infusion levels after 80% of the induction-abortion interval had elapsed. The main decline in estrogen levels occurred in individual women either during the 1st quarter or during the last quarter of the induction-abortion interval. There were no significant relationships between changes in estrogen levels and the interval from 1st administration of PGF2a to subsequent abortion.

Studies of human chorionic gonadotrophin, placental lactogen, and progesterone secretion in vitro

The relative ability of explants of villous tissue from 8- to 10-week placenta to synthesize and release human chorionic gonadotrophin (HCG), human placental lactogen (HPL) and progesterone in culture was investigated. It was found that the synthesis and release of all three hormones was two to four times greater than the initial tissue content in the first 24 hours in culture. Between 24 and 48 hours of the secretion of HCG, HPL and progesterone was reduced to about 40 per cent, 20 per cent and 60 per cent respectively of that produced in the previous 24 hours. These results support previous findings that the production of HPL is more labile than HCG in culture, and suggest that progesterone secretion under the present conditions is least affected. When azastene (4, 4, 17 alpha-trimethylandrost-5-eno (2,3-d) isoxazol-17-ol), an inhibitor of gonadal and placental 3 beta-hydroxy steroid dehydrogenase conversion of pregnenolone to progesterone in rhesus monkeys, was added to the culture medium at a concentration of 0.3-150 mumol/l, no marked effect on the secretion of progesterone was observed. This indicates that the dehydrogenase in human placental tissue is either less accessible to azastene or that the inhibitory activity in the rhesus monkey was due to a metabolite of azastene.

Two Cases of Placental Sulphatase Deficiency with Unusual Endocrine Features

Two cases of placental sulphatase deficiency are described both of which showed typical clinical features, associated with very low urinary and serum oestrogens. In the one case that reached term there was a spontaneous rise in serum oestradiol levels (from 16 to 44 nmol/1) after 38 weeks, although no oestradiol rise occurred after DHEAS loading. She later achieved a vaginal delivery. Although no steroid sulphatase activity could be detected in the placenta of case 2, there was a small transient rise in serum oestradiol (from 15.4 to 32.7 nmol/I) after maternal DHEAS loading. However, the absolute levels reached were very low compared to the response seen in normal pregnancies. The spontaneous rise in basal oestradiol levels between 38 and 41 weeks and the transient rise following DHEAS loading have not been previously reported for patients with placental sulphatase deficiency. Umbilical cord blood levels of several steroids in case 2 showed some similarities to previous case reports. Levels of DHEA, 16αOH DHEA, 170H pregnenolone and Cortisol were significantly lower than control values. DHEAS, 16αOH DHEAS, 170H progesterone and andros‐tenedione were within the normal range.

The secretion of hormones during the culture of human preimplantation embryos with corona cells**Supported by the National Health and Medical Research Council of Australia.

Following in vitro fertilization of human preovulatory oocytes from spontaneously ovulating women, determinations were made of the secretion into the culture medium of progesterone, estradiol, human chorionic gonadotropin (hCG), and the prostaglandins (PG) E2 and F2 alpha, over a 3- to 4-day period of embryo development. It was found that the corona cells associated with the egg could account, between days 2 and 3, for a mean daily secretion of 50 ng of progesterone and approximately 100 pg of estradiol, PGE2, and PGF2 alpha, respectively. Mechanical removal of the corona cells after about 48 hours for the examination of the egg for cleavage reduced the mean amount of progesterone produced on day 3 to 2 ng, and a concomitant decrease in estradiol. PGE2, and PGF2 alpha was observed. Steroid secretion could be restored on day 3 to more than 50% of that secreted on day 2 by returning some detached corona cells to the culture medium containing the embryo. hCG was not detected (less than 2 mIU/ml) in the culture medium at any stage of embryo culture over the 3- to 4-day period.