Evidence for progesterone synthesis by human umbilical cord blood erythrocytes.

In order to determine whether human umbilical cord blood erythrocytes are capable of progesterone biosynthesis, sonicated preparations of plasma-free erythrocytes at the range of total cell numbers between 18.4 x 10(9) and 62.9 x 10(9) cells obtained from umbilical cord arterial and venous blood collected from normal pregnant women (n = 6, age 28-39 years) following spontaneous vaginal delivery were incubated with [7n-(3)H]-pregnenolone as substrate. The leucocyte content of the incubates was negligible (<0. 005%). Controls (n = 4, age 29-36 years; 27.6 x 10(9) to 42.7 x 10(9) erythrocytes) obtained from cord blood of normal pregnant women were heat-denatured preparations of cells. Using the reverse-isotope dilution technique, [(3)H]-progesterone was isolated and characterized yielding an overall enzymic conversion which ranged between 0.27 and 0.46%. The results indicate for the first time that cord blood erythrocytes possess the 3beta-hydroxysteroid dehydrogenase-5,4-en isomerase activity and are a source of progesterone in human pregnancy.

Expression of C-20, 22 desmolase activity by the human fallopian tube in vitro: evidence for steroidogenesis.

With a view to establishing whether cells of the human Fallopian tubes possess the cholesterol side-chain cleavage activity, homogenates of the tubes, obtained from 6 women (39-45 years) following abdominal hysterectomy for benign conditions, were incubated with (7n-3H)-cholesterol as substrate. Controls (n=6, age 40-44 years) were homogenates heated in a boiling water bath for 10 min. Using the reverse-isotope dilution technique, (3H)-pregnenolone was isolated and characterized. No such metabolite was evident in control incubations of heat-denatured enzymes. The extent of enzymic conversion varied from 1.9 x 10(-3) to 1.3 x 10(-2)%. The results reveal for the first time the existence of an active cholesterol-specific C-20, 22 desmolase system in the viable tissues. It is suggested that there exists a potential for substantial pregnenolone synthesis in vivo. This rate-limiting steroid biosynthetic conversion provides a new dimension to the functional capacity of the Fallopian tubes in the synthesis of steroids, which may be necessary for modulating ciliary beat frequency and in maintenance of hormonal milieu essential for embryogenesis.

A delta 4-3-keto pathway for testosterone synthesis in the human spermatozoa.

The ability of human spermatozoa to metabolize pregnenolone to progesterone and progesterone to testosterone was assessed. Sonicated specimens of freshly ejaculated sperm from two groups of husbands (n = 6, age 32-38 years; n = 6, age 30-51 years) of infertile couples in the range of sperm concentration between 237.5 and 568.5, 100.1 and 248.8 millions per ejaculate, were separately incubated with [7n-3H]pregnenolone and [1,2,6,7,16,17-3H]progesterone. Using the classical reverse-isotope dilution technique the desired products [3H]progesterone and [3H]testosterone formed from the respective substrates were isolated and characterized, yielding 1.4 to 12.2% and 3.1 x 10(-2) to 2.0 x 10(-1)%. Such metabolites were not evident in the controls. The results indicate that the human spermatozoa contain the enzymes necessary for the transformation of pregnenolone to testosterone via the delta 4-3-keto route.

Characterization of a delta 5-3 beta-hydroxy pathway of testosterone synthesis in the human spermatozoa.

Sonicated specimens of freshly ejaculated sperm from two groups of husbands (n = 6, age 32-38 years; n = 7, age 27-38 years) of infertile couples in the range of sperm concentration between 237.5 and 568.5; 131.1 and 256.7 millions per ejaculate were separately incubated with [7n-3H]pregnenolone and [1,2,6,7(-3)H]dehydroepiandrosterone. Using the reverse-isotope dilution technique [3H]dehydroepiandrosterone and [3H]testosterone formed from the respective substrates were isolated and characterized, yielding 1.2 x 10(-2) to 4.6 x 10(-20/0) and 7.1 x 10(-2) to 2.5 x 10(-10/0)/ Such metabolites were not evident in the controls. These results provide evidence for metabolic transformation of pregnenolone to testosterone via the delta 5-3 beta-hydroxy route.

A comparative study of progesterone synthesis by term human fetal amnion and chorion.

Activity of 3 beta-hydroxysteroid dehydrogenase-5, 4-en isomerase was investigated with [7n-3H] pregnenolone as substrate in the homogenates of amnion and chorion collected from women (n = 6, age 20-34 years) after spontaneous labour at term (37-40 weeks gestation) from uncomplicated pregnancies. Controls were heat-denatured homogenates instead of fresh homogenates. Reverse-isotope dilution analysis gave positive identification of [3H] progesterone in all incubations of viable tissues. This metabolite was not evident in the controls. The extent of enzymic conversion of chorion, 1.4-31.1% was consistently higher than that of the amnion, 0.66-1.7%. The results show that the human fetal membranes can be a source of progesterone during pregnancy. The metabolic capability may have physiological significance in the induction and progression of labour to parturition.

A relationship between sperm count and intracellular progesterone, 17 alpha-hydroxyprogesterone, and cortisol concentrations in human spermatozoa.

In an attempt to determine whether human spermatozoa contain steroid hormones, in this study we have measured the immunoreactive levels of P, 17 alpha-OHP, and F in the extracts of sonicated specimens of sperms obtained as ejaculates from husbands of infertile couples. The results of this study demonstrate for the first time that human spermatozoa contain definite amounts of the measured steroids and suggests an association between steroid levels and sperm count. These steroids identified intracellularly in the haploid germ cells may play a role in the molecular events associated with fertilization.

Concentrations of intracellular sex steroids in human spermatozoa.

In an attempt to determine whether human spermatozoa contain steroid hormones, the immunoreactive levels of progesterone, testosterone, and estradiol-17 beta were measured in the extracts of sonicated specimens of sperm obtained as ejaculates from husbands of infertile couples (n = 67, age 26-44 years). In the range of sperm concentration between 3.0 and 869.6 million per ejaculate (143.2 +/- 13.6 mean +/- SEM), the concentrations were as follows: progesterone 127.3-6685.4 fmol/ml (1434.5 +/- 178.6), testosterone 98.3-2219.1 fmol/ml (611.6 +/- 49.0), and estradiol-17 beta 33.0-678.1 fmol/ml (206.9 +/- 19.7). Using Pearson's correlation coefficient, the steroid hormone levels correlated significantly with the total sperm number (p < .02). The spermatozoa contained small but measurable amounts of the sex steroids.

Response of ovary in young women experiencing laparoscopy under general anaesthesia.

This study investigated whether changes in circulating levels of immunoreactive oestradiol-17 beta (E2), progesterone (P) and testosterone (T) occur in women at follicular (n = 18, age 25 to 39 years) and luteal (n = 17, 25 to 39 years) phases of the normal menstrual cycles, experiencing laparoscopy after intravenous sedation with general anaesthesia. The pre- and intra-operative follicular phase plasma steroid hormone concentrations were 153.5 +/- 84.3 vs 297.4 +/- 220.8 pg/ml for E2, 2.0 +/- 3.2 vs 3.3 +/- 3.8 ng/ml for P and 746.6 +/- 415.9 vs 1325.8 +/- 535.1 pg/ml for T, respectively. The corresponding luteal phase steroid levels were 259.7 +/- 120.2 vs 382.7 +/- 188.7 pg/ml, 7.0 +/- 4.8 vs 9.9 +/- 6.1 ng/ml and 819.4 +/- 355.7 vs 1703.5 +/- 1058.1 pg/ml. Using the Wilcoxon rank sum test, intra-operative hormone levels with the exception of P in the luteal phase were found to be significantly elevated (p < 0.05). The results suggest that laparoscopy under general anaesthesia evokes increased secretion of ovarian hormones, possibly via the activation of hypothalamo-pituitary-ovarian axis.

In vitro formation of testosterone via the delta 5-pathway in the human umbilical cord.

Homogenates of two groups of term umbilical cord (n = 6, 37-40 weeks; n = 6, 38-40 weeks) were separately incubated with [7n-3H]pregnenolone and [1,2,6,7-3H]dehydroepiandrosterone. Using the reverse-isotope dilution technique, [3H]dehydroepiandrosterone and [3H]testosterone formed from the respective substrates were isolated and characterized. The extent of enzymic conversions were 0.015-0.28% and 0.044-2.2%. These results provide evidence for the metabolic transformation of pregnenolone to testosterone via the delta 5-3 beta-hydroxy route.

Human umbilical cord as a possible source of progesterone.

Homogenates of human term umbilical cord were incubated with 7n-3H-pregnenolone. Using a reverse-isotope dilution technique, 3H-progesterone was isolated and characterized. The extent of enzymic conversion ranged from 2.5 to 55.4%. The results indicate that the cord can be a source of progesterone during human pregnancy.

Elevated peritoneal fluid luteinizing hormone and prolactin concentrations in infertile women with endometriosis.

In this study, we compared (Mann-Whitney U-test) the peritoneal fluid FSH, LH and PRL levels, measured by RIA, at the follicular and luteal phases of the menstrual cycle in women with (n = 43; age 25-44 years) and with no evidence of endometriosis (n = 35; age 25-39 years) who were considered as controls. Both follicular and luteal phase FSH concentrations of women with endometriosis were not statistically different (n = 22 vs 18; 0.32-5.8 vs 0.50-8.2 IU/l, P = 0.247; n = 13 vs 14; 0.6-6.5 vs 0.66-6.7 IU/l, P = 0.604) compared to their respective controls. In contrast to FSH, the concentrations of LH at follicular (n = 19 vs 17; 3.1-34.2 vs 2.3-12.2 IU/l, P = 0.01) and luteal (n = 17 vs 15; 2.1-95.4 vs 1.3-17.9 IU/l, P = 0.02) phases of the test group was significantly elevated at both phases of the cycle. With respect to differences in PRL concentrations at follicular phase no significant change (n = 21 vs 16; 1030-5800 vs 1305-4650 mIU/l; P = 0.255) was observed. The greatest difference in luteal PRL concentrations (P = 0.007) was obtained between the women with endometriosis and controls (n = 17 vs 17; 1895-8600 vs 1041-5000 mIU/l). The results suggest that disordered synchronization of neuroendocrine mechanisms controlling LH and PRL may be the underlying abnormality causing infertility in our group of patients with endometriosis.

Cholesterol side-chain cleavage activity in the human umbilical cord in vitro.

With a view to establish whether cells of the human umbilical cord possess cholesterol side-chain cleavage activity, homogenates of term umbilical cord obtained following spontaneous vaginal delivery from uncomplicated pregnancies (n = 6; 38-40 weeks gestation) were incubated with 26-14C cholesterol. Controls were homogenates heating in a boiling water bath for 10 min. Proof of the existence of cholesterol C-20, 22-desmolase activity in the viable tissue was established by associating radioactivity due to the labelled carbon 26 of 26-14C cholesterol with the p-bromophenacyl ester of isocaproic acid by reverse-isotope dilution analysis. The desmolase efficiency expressed as specific activity of 14C isocaproic acid, dpm g-1 tissue varied from 110 to 351 with a mean of 202. The small but definite conversion indicates that the cholesterol side-chain cleavage reaction previously believed to be exclusive to the steroidogenic pathway operating in the adrenal, gonads and placenta does exist in the human term umbilical cord in vitro.

Upper genital tract fluid oestradiol-17 beta, progesterone and testosterone concentrations during the menstrual cycle.

This study was to obtain evidence whether a cyclical pattern in the concentrations of oestradiol-17 beta, progesterone and testosterone exists in the upper genital tract fluid of women with normal menstrual cycles. The steroid concentrations at proliferative and luteal phases were n = 20 vs 17, 54.5-190.0 vs 44.2-827.1 pg/ml; 0.27-3.3 vs 3.4-46.5 ng/ml and 41.9-734.8 vs 85.7-991.2 pg/ml, respectively. The results were analysed according to Mann-Whitney U test. Significantly higher concentrations of oestradiol-17 beta (p = 0.0036) and progesterone (p = 0.0001) were found. The results suggest the existence of a cyclical variation and point to the ovary as the main source of output of the steroids.

A comparative study of peritoneal fluid concentrations of oestradiol-17 beta, progesterone, and testosterone in controls and in patients with endometriosis.

To ascertain whether a cyclical variation exists in the steroid hormone concentration of the peritoneal fluid between controls and patients with endometriosis, we set out to measure the concentration of oestradiol--17 beta (E2), progesterone (P4), and testosterone (T) by specific radioimmunoassay in the peritoneal fluid of these 2 groups. Forty six patients (mean age, 30.1 +/- 6 years) with pelvic endometriosis and 32 control patients (mean age, 32.2 + 3.8 years) without endometriosis were studied. The significance of differences between groups was determined by the Mann-Whitney mu test. In the control group a significant rise in peritoneal fluid E2 (P = 0.017) and P4 (P = 0.048) levels was observed in the luteal phase compared to the follicular phase, but no such difference was noted in women with endometriosis. It is suggested that peritoneal fluid steroid hormone assays are not helpful in the evaluation of ovarian function in endometriosis.

Control of gonadotrophin secretion by steroid hormones in castrated male transsexuals. I. Effects of oestradiol infusion on plasma levels of follicle-stimulating hormone and luteinizing hormone.

Twenty-four infusions of oestradiol (E2) in graded doses ranging from 0--200 micrograms administered over a period of 7 hours were carried out in eleven healthy male transsexuals who had undergone sex reassignment at least 3 months previously. Plasma levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH) and E2 were analysed by radioimmunoassay, while sex hormone binding globulin (SHBG) was measured using 3H-testosterone as the saturating ligand. Infusion of 5--200 micrograms of E2 raised plasma E2 to levels ranging from 38.4--367.8 pg/ml which present 83%--800% of levels found in a control group of sixty-three normal males. SHBG capacity remained unchanged at all doses of E2 studied. No change in plasma levels of FSH and LH was observed in control infusions and infusion of 5 micrograms of E2. From 10 micrograms-200 micrograms, suppression of plasma levels of FSH was noted at the 5--7 hour period. The suppression increased up to 20 micrograms and thereafter the levels of FSH remained constant. On the other hand, the suppression of LH increased up to the highest E2 dose (200 micrograms) studied. Further, significant suppression of LH occurred earlier than the 5--7 hours as the dose of E2 increased. These observations are consistent with the conclusions that: (1) E2 plays a part in the regulation of secretion of FSH and LH in men; and (2) at doses higher than physiological, E2 exerts a differential effect on the secretion of FSH and LH.

The effect of clomiphene on uncastrated and castrated male transsexuals.

Clomiphene was administered at a daily dose of 100 mg for 5 consecutive days to two groups of male subjects. There were three uncastrated male transsexuals (Group A) and nine castrated male transsexuals (Group B). In the uncastrated subjects Clomiphene enhanced the secretion of follicle-stimulating hormone (FSH), luteinizing hormone (LH) and testosterone (T) starting from day 3 of treatment. The magnitude of the increment ranged from 150%--250% over the mean basal plasma level for LH, 113%--262% for FSH and 12%--68% for T. In contrast, Clomiphene suppressed both FSH and LH in the castrated subjects. Equal suppression of both the gonadotrophins were observed. The levels of FSH and LH in plasma on day 3 were suppressed to 66.4% and 66.2% of their respective mean basal levels. Two days after the termination of treatment, suppressions were maintained. These results suggest that Clomiphene acts as an anti-oestrogen in uncastrated male transsexuals and in castrated male transsexuals who have subphysiological levels of oestradiol it acts as a weak oestrogen.