Samples of myometrium were obtained from twenty-two patients undergoing caesarian sections for various obstetric indications. The myometrium was homogenized and divided into four subcellular fractions (crude nuclear, mitochondrial, microsomal and cytosol). The concentration of endogenous progesterone in these fractions was determined by radioimmunoassay. In addition the concentration of oestrone and oestradiol of fifteen cytosolic fractions was determined. The subcellular concentration of progesterone (pg/mg protein) in human myometrium was higher in samples taken during labour than in samples taken at elective caesarian sections. This finding was statistically significant in the crude nuclear (P less than 0.005) and the microsomal (P less than 0.05) fractions. The absolute concentration of progesterone was lowest in the nuclear and highest in the microsomal fraction. The relative progesterone concentrations in the four subcellular fractions were the same in both groups. The data show that there is no significant decrease in myometrial progesterone associated with labour in man. The mean concentration of oestrone was higher than the mean oestradiol concentration in the cytosol fraction of human myometrium in late pregnancy. This study shows that a completely different oestrogen ratio exists in myometrium than in plasma.

Homogenates of cerebral metastatic chorionepithelioma tissue were incubated with labelled dehydroepiandrosterone, pregnenolone or 20alpha-hydroxypregn-4-en-3-one. The metabolites of dehydroepiandrosterone which were isolated and identified were androstenedione, testosterone, oestrone, and oestradiol; no oestriol was detected. The only metabolite of pregnenolone and 20alpha-hydroxypregn-4-en-3-one isolated and identified was progesterone. No conversion of C-21 to C-19 steroids occurred in the metastatic chorionepithelioma tissue.

Six sexual steroidal hormones (progesterone, pregnenolone, testosterone, ethinyl-oestradiol, oestriol and oestrone) inhibit acetylcholine- and histamine-induced contractions of the isolated guinea-pig ileum. Different degrees of inhibitory capacity were found. High concentrations of PGE1 and F2alpha reverse some of these inhibitions. This reversal seems to be non-specific and probably related to sensitization of the ileal smooth muscle by prostaglandins. The inhibitory action of sexual steroids might be non-specific as well. But a "corticoid-like" effect cannot be excluded.

Gilts were treated on Day 112 of gestation with saline or a prostaglandin (PG) F-2 alpha analogue. In control gilts there was a rise in the relaxin concentration from 48 h before the onset of delivery, peaking between 12 and 28 h pre partum followed by a steep fall. The relaxin concentrations at each 1-h time interval were analysed in relation to the farrowing interval for each gilt using correlation analysis. There was a significant (at least P less than 0.05) positive correlation between the relaxin concentration and the farrowing interval at every time period from 14 to 2 h before delivery. In contrast there was little relationship between concentrations of progesterone, oestrone and oestradiol-17 beta and farrowing intervals. The gilts treated with PGF-2 alpha analogues had steroid profiles indistinguishable from those in controls but differing relaxin secretion patterns. Relaxin concentrations peaked at 1-2 h after PGF-2 alpha injection and this was followed by a second smaller increase closer to the time of delivery in 7 out of 12 gilts. The 'two-peak' gilts had significantly higher relaxin concentrations at farrowing and took significantly longer to farrow than did the 'one-peak' gilts (P less than 0.005). These results suggest that high relaxin concentrations during the last 14 h before the onset of parturition are associated with increased farrowing times, but are not associated with any increase in neonatal mortality.

Prostaglandin F (PGF) was measured in amniotic fluid, and 13,14-dihydro-15-keto-prostaglandin F2 alpha (PGFM) was measured in maternal peripheral venous plasma and amniotic fluid of rhesus monkeys during late pregnancy. 13,14-Dihydro-15-keto-PGF2 alpha was determined in the maternal peripheral venous plasma of two animals following intra-uterine foetal death. The mean concentration of PGF and PGFM in amniotic fluid increased fourfold during the last 5 days of pregnancy. This increase was associated with an increase in the oestrone concentration in amniotic fluid and in maternal plasma. In normal pregnancy there was no increase in PGFM levels in the maternal peripheral vein, up to 1-2 days pre partum. After intra-uterine death, progesterone concentration in the maternal peripheral vein was unaltered, but oestrone and oestradiol declined. In plasma samples taken within 12 h of delivery, the concentration of PGFM was raised. It is concluded that an increase in prostaglandin production accompanies delivery at normal term, and at delivery past term following intra-uterine foetal death.

In this paper we examine the ability of the mammary gland to remove from circulating blood three compounds which differ in their physico-chemical and structural properties. Mammary extraction of progesterone, oestrone sulphate and epidermal growth factor (EGF) is similar at peak lactation in goats, but the proportion of labelled infusate that is transferred into milk is greater for oestrone sulphate and EGF than progesterone which is rapidly metabolised by mammary tissue. The kinetics of transfer of progesterone, oestrone sulphate and EGF from blood into milk show that transcellular processes are involved, and on the basis of earlier hypotheses and new information reported here the results indicate the probable importance of simple and facilitated diffusion pathways for progesterone and oestrone sulphate, and secretory mechanisms for oestrone sulphate and EGF. Although evidence is lacking for a direct effect of hormones in milk on mammary function, their concentration in milk may reflect changes in local regulation of mammary secretion. Considerable practical value is attached to the immunodiagnostic use of milk hormone concentrations to determine ovarian and placental endocrine activity during pregnancy in domestic ruminants.

Ovarian homogenates from 10-, 23- 128- and 60-day-old golden hamsters were incubated with [14C]-4-androstene-3,17-dione or [7-3H]-progesterone in the presence of NADPH and enzyme activities and metabolism of progesterone were estimated. A rapid increase in uterine weight was found around 28 days of age. The activity of 5 alpha-reductase was very high in the ovaries of 23-day-old hamsters (647 +/- 117 (SD) nmol/g tissue/h), high in those of 28-day-old hamsters (135 +/- 4) and low in those of 10- and 60-day-old animals (20 +/- 16, 39 +/- 11). However, the activity of 5 beta-reductase was high in all ovaries of golden hamsters at different stages of development (84-132 nmol/g tissue/h). The major C-21-17-hydroxysteroids and C19-steroids formed from progesterone by the ovaries of 23-day-old hamsters were 5 alpha-steroids such as 3 alpha, 17-dihydroxy-5 alpha-pregnan-20-one and androsterone, whereas those by the ovaries of 28- and 60-day-old hamsters were 4-ene-3-ketosteroids and 5 beta- and 5 alpha-steroids such as 17-hydroxy-4-pregnene-3,20-dione. 3 alpha, 17-dihydroxy-5 alpha- and 5 beta-pregnan-20-one, 4-androstene-3,17-dione, testosterone and androsterone. The formation of oestradiol-17 beta and oestrone from progesterone was found only in the ovaries of 38- and 60-day-old hamsters. These results show that similarly high levels of 5 beta-reductase are present in all ovaries from suckling, immature and adult golden hamsters and that high levels of 5 alpha-reductase are formed only in ovaries from immature hamsters, especially those with small uterus. The active 5 alpha-reduction of 4-ene-3-ketosteroids may be responsible for the decrease in the formation of oestrogens in immature hamster ovaries.

The endocrine regulation of parturition exhibits several interspecies differences. Among the endocrine parameters considered to be the most important ones from the point of view of the regulation of parturition the concentration of oestrogens (oestradiol, oestriol and oestrone), progesterone, PGF2 alfa and its degradation product PGFM, was determined in both peripheral and uterine venous blood by RIA method in rats starting from the 15th day of pregnancy to the 3rd or 4th day following parturition. In the pregnant rat oestriol could only be detected on the day of parturition (21th day). Concentrations of oestriol and oestradiol were the highest on the day of parturition. An opposite tendency could be observed as far as progesterone concentration was concerned, i.e., the concentration decreased gradually from the 15th day of pregnancy onwards and the lowest value was reached on the day of parturition both in peripheral and uterine venous blood. PGF2 alfa and PGFM concentrations in the uterine vein increased gradually from the 15th day of pregnancy and the highest value could be detected peri partum. Our data, in accordance with those of others support the idea that the hormones investigated are involved in the process of parturition, i.e., characteristic changes of oestrogens, progesterone and prostaglandin levels ensue in the rat prior to parturition.

This study was designed to assess the effect of an altered level of serum oestrogen and progesterone on the prolactin (PRL) response to gonadotrophin releasing hormone (GnRH). Six normal women were studied in the early follicular phase and the mid-luteal phase of one cycle and five menopausal women were studied before and after treatment with progesterone. Blood samples were collected at 15 min intervals for 6 h after a basal collection period of 30 min. Intravenous boluses of GnRH (1 microgram, 10 micrograms and 50 micrograms) were given at 0, 2 and 4 h. Basal samples were assayed for 17 beta-oestradiol (E2), oestrone (E1) and progesterone (P); LH, FSH and PRL were measured in all samples. Serum PRL was significantly elevated in all groups after 10 micrograms of GnRH with maximum increments (+/- SEM) ranging from 3.9 +/- 1.3 micrograms/l in early follicular phase women to 14.7 +/- 4.7 micrograms/l in progesterone-treated menopausal women. The PRL response to GnRH was significantly greater in the luteal phase and in menopausal women compared to early follicular phase women. There was a significant correlation between the maximum PRL response and the maximum LH response to GnRH in all the women studied (r = 0.7; P less than 0.01). A significant correlation was also found between the maximum PRL response and the basal serum oestrogen concentration in the normal cycling women (r = 0.8; P less than 0.01), but not when the menopausal women were included in the analysis.(ABSTRACT TRUNCATED AT 250 WORDS)

Progesterone, androstenedione, testosterone, oestrone and oestradiol were measured in the postovulatory follicle (POF) at various times up to 52 h after ovulation. The 3-fold decrease in progesterone, the major constituent, which occurred over the first 15--20 h resembled the changes previously described for the enzyme, 3beta-hydroxy-steroid dehydrogenase. At 1 h before ovulation the granulosa cells of the anteovulatory follicle (AOF) contained 50 times more progesterone than the POF granulosa fraction collected 2--3 h later. The thecal portion of the AOF had progesterone concentrations 5 times those of the POF theca, but the latter contained higher concentrations of androstenedione and oestrone.

We have studied the binding of a number of radiolabelled steroids and lipophilic substances to porcine corpus luteum (CL) particulate fractions. Following preincubation of CL homogenates with radiolabelled progesterone or pregnenolone prior to fractionation on continuous sucrose density gradients, a broad peak of binding was observed associated with a particulate fraction of buoyant density 1.05-1.10 g/cm3. Progesterone content also peaked at a similar buoyant density (1.06-1.12 g/cm3). Pretreatment of luteal homogenates with digitonin perturbed the buoyant density of the progesterone-binding particulate fraction to 1.10-1.14 g/cm3 and sharpened the binding peak. Progesterone content was also perturbed to a similar extent by digitonin pretreatment, without release of the steroid. Oestrogens were also sequestered by this fraction, but steroid precursors (cholesterol, cholesterol ester), corticosteroids (cortisol, corticosterone), sterol conjugates (oestrone sulphate, pregnanediol glucuronide) and other lipophilic substances (arachidonic acid, phospholipid, prostaglandins E1, E2 and F2 alpha) were not bound. Androgens were bound weakly by fractions from control gradients but, in the presence of digitonin, significant binding could be demonstrated. Radiolabelled steroids were shown to interact directly with luteal membrane fractions, rather than interacting first with cytosolic steroid receptors which then bound to membranes. Furthermore, [3H]progesterone was not bound by porcine granulosa cell particulate fractions. These observations suggest that this fraction may be involved in sequestration or packaging of progesterone for secretion by the luteal cell.

Granulosa cell responsiveness at an early (1-2 h) or late (14-16 h) stage of differentiation following the onset of oestrus [and presumably the LH surge] was studied in 16 cows. Follicular fluid collected at the early stage (8 preovulatory follicles) had a higher concentration of testosterone (P less than 0.05), oestradiol (P less than 0.01) and oestrone (P less than 0.01) than did follicular fluid collected at the late stage of oestrus (8 preovulatory follicles). No difference in follicular fluid progesterone was noted between follicles collected at the early and late stages of oestrus. Granulosa cells collected at the early stage of oestrus had a higher in-vitro response (progesterone production) to LH (P less than 0.05), forskolin (P less than 0.08) and diacylglycerol (P less than 0.05) than did granulosa cells collected at the late stage of oestrus. However, later stage granulosa cells produced more (P less than 0.01) progesterone after culture with prostaglandin E-2 than did earlier stage granulosa cells. These results show that follicular fluid oestrogen decreases, which suggests a loss of aromatase activity as oestrus progresses, and that granulosa cells become refractory (low progesterone production) to in-vitro LH, forskolin, and diacylglycerol challenge, yet acquire responsiveness to prostaglandin E-2 as oestrus progresses.

We have described previously the presence of binding sites in particulate fractions of the porcine corpus luteum (CL) which were specific for progesterone. We now demonstrate the presence of similar progesterone-specific binding sites in particulate fractions of the ovine CL. Preincubation of ovine luteal membranes with radiolabelled steroids demonstrated binding of progesterone and pregnenolone to a low-density particulate fraction (1.07-1.09 g/cm3). Preincubation with digitonin perturbed the buoyant density of this fraction (to 1.10-1.14 g/cm3) without causing release of steroid. Androgens and oestrogens did not bind appreciably to control luteal membranes, but were bound when preincubated with digitonin. In contrast, steroid conjugates (oestrone sulfate, pregnanediol glucuronide), cortisol, fatty acids (arachidonic acid, prostaglandin F2 alpha) and cholesterol ester failed to bind to ovine luteal membranes, with or without digitonin pretreatment. The effects of digitonin on steroid binding led us to examine its effects on steroid binding to ovine luteal membrane fractions in vitro. Specific progesterone binding was absent in the absence of digitonin, even at very high membrane concentrations. However, binding of 3H-labelled progesterone was stimulated 5-15-fold in a dose-dependent fashion by increasing digitonin concentrations, reaching a plateau at about 100 microM. In the presence of digitonin, [3H]progesterone binding increased linearly with luteal membrane concentration. Other detergents, saponins and cardiotonic steroids tested did not stimulate progesterone binding to ovine luteal membranes. [3H]Progesterone binding was dependent on the pH, duration and temperature of incubation. Unlabelled progesterone decreased binding of [3H]progesterone (half-maximal displacement of specific binding (IC50) at about 60 nM) whereas androgens were less potent (IC50, 500-3300 nM), whilst a wide range of other steroids and inhibitors of steroidogenic enzymes were ineffective, except at very high concentrations. Similarly, a number of progesterone receptor agonist and antagonist analogues failed to compete for progesterone binding to luteal membranes, suggesting that these binding sites were unrelated to progesterone receptors. Modifications to the 3, 4, 5 and 11 positions of progesterone, removal of the steroid side-chain or aromatization of the A-ring decreased binding potency dramatically, whereas changes to the 17 or 20 positions had relatively minor effects. Our results indicate the presence of a low density particulate fraction in ovine corpora lutea which contains specific binding sites for endogenous and exogenous progesterone.

This study was designed to evaluate whether traditional plasma hormone determinations can be adequately replaced by measurements of salivary hormones. Eleven young sportswomen with menstrual irregularities attributed to strenuous physical exercise participated in this study. Mean body weight expressed as a percentage of ideal body weight was 92%, SD 4%. Their mean weekly training distance was 35 km, SD 15. Basal plasma endocrinological measurements revealed a hypo-oestrogenic status (mean plasma oestradiol values: 22 pg.ml-1, SD 8.8), and a deficient luteal phase (mean plasma progesterone: 2.9 ng.ml-1, SD 2.1). Pre-exercise salivary sex steroids were low. Salivary progesterone levels were 39.3 pg.ml-1, SD 9.5 (normal ranges in saliva: 25-60 pg.ml-1), salivary oestrone (E1) was 12.2 pg.ml-1, SD 2.3 (normal ranges in saliva: 7.5-25 pg.ml-1), and salivary oestradiol (E2) less than 1.9 pg.ml-1, SD 1.1 (normally 1.0-10.0 pg.ml-1). After a 21-km run, all salivary steroids appeared to increase. Mean salivary testosterone levels increased by 15.2% and salivary progesterone by 14.8%. Mean salivary oestrogens also increased (E1: +13.9%; E2: +21.1%). These findings confirm the results of earlier studies which found higher post-exercise plasma sex steroid levels. Since salivary measurements are believed to reflect non-protein-bound, thus free steroid levels, the results obtained by these techniques may provide a more realistic picture of the hormonal effects of physical exercise. In future, more accurate, cost-effective and easier techniques for salivary measurements may offer additional advantages.

Follicular growth and hormone determinations were used to predict ovulation in the mare. Thirty Finnhorse mares were used for the investigation and 38 oestrous cycles were studied. The mares were examined by rectal palpation and ultrasonography every 6 hours during late oestrus. Daily blood samples were obtained for progesterone and oestrone sulphate determination. The preovulatory follicle grew 3 mm a day up to 2 days before ovulation. The size then remained constant, before diminishing by 2-3 mm during the last 12 hours. The maximal diameter of the follicle was 43 +/- 4 mm. In 89% of the follicles the round shape became more irregular before ovulation. During the last 24 hours before ovulation 37 of 38 follicles were regarded as mature on rectal palpation. The oestrone sulphate level was highest 24-48 hours before ovulation, the first decrease being observed most commonly around ovulation (+/- 1 day). The size of the follicle was the most reliable criterion in the prediction of ovulation.

Plasma concentrations of 15-ketodihydroprostaglandin (PG) F2 alpha, progesterone, oestrone sulphate, oestradiol-17 beta and cortisol during late gestation, parturition and the early post-partum period were measured in six llamas and five alpacas. During the last 100 days of pregnancy, 15-ketodihydro-PGF2 alpha concentrations increased steadily until the day of parturition when a massive release was detected (P < 0.01) concomitant with a decrease in progesterone concentrations (P < 0.01). The highest PGF2 alpha metabolite concentrations (159 +/- 35 nmol l-1 and 92 +/- 29 nmol l-1 in llamas and alpacas respectively) were detected in the sample collected during the morning on the day of parturition. Basal concentrations were registered by day 3 after delivery. Plasma concentrations of oestrone sulphate started to increase 80 days before parturition and reached peak concentrations immediately before parturition (15 +/- 3 nmol l-1 in llamas and 18 +/- 5 nmol l-1 in alpacas). Oestrone sulphate concentrations dropped sharply (P < 0.01) on the day of parturition in llamas and one day later in alpacas, whereupon they remained relatively unchanged until at least 20 days postpartum. Oestradiol-17 beta concentrations were higher than 180 pmol l-1 during the last 45 days of pregnancy, began to decrease on the day of parturition and reached very low concentrations within the following two days. High oestradiol-17 beta concentrations were registered 7 days postpartum in all alpacas (P < 0.05) and within 10 days of parturition in five of six llamas (P < 0.01). No significant cortisol peaks were observed around parturition, but mean concentrations were increased in both species.

Basal body temperature (BBT) was measured continuously by radiotelemetry throughout 14 chimpanzee menstrual cycles and correlated with daily observations of the sexual skin swelling. A biphasic BBT shift from a pre-nadir mean of 36-12 degrees C to a post-nadir mean of 36-67 degrees C was observed in 12 cycles. The temperature nadir showed a close temporal relationship with detumescence of the sexual skin swelling (an early luteal event), but the rate of temperature rise after the nadir was variable. In 3 normal cycles studied, the temperature nadir occurred the day after a urinary oestrone peak, but there was no consistent temporal association between BBT rise and pregnanediol increment. Progesterone secretion is therefore probably not the sole determinant of the BBT shift; the changing oestrogen/progestin ratio may be the more important factor regulating body temperature during the luteal phase.

The comparative effect of oestradiol-17 beta, oestrone and oestrone-3-sulphate was examined on guinea-pig endometrium in primary culture. A parallel study was conducted in vivo to appreciate hormonal effects on the uterine luminal surface of ovariectomized guinea-pigs. Scanning electron microscopy studies showed that uterine epithelial cells were responsive to physiological concentrations of E2, E1 and E1S. Their plasma membrane was dramatically modified by 2.10(-9) M E2 and 10(-7) M E1S, but there are clear qualitative differences between the effect of E2 and of E1S. These effects were abolished by progesterone or 4-hydroxy-tamoxifen. Guinea-pig uterine epithelial cells in primary culture and in vivo are responsive to E2 and to E1S but their response to E1S appears to be more specific and can be distinguished by the cell surface morphology.