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Purpose: There is strong epidemiologic evidence indicating that estrogens may not be the sole steroid drivers of breast cancer. We hypothesize that abundant adrenal androgenic steroid precursors, acting via the androgen receptor (AR), promote an endocrine-resistant breast cancer phenotype., Experimental Design: AR was evaluated in a primary breast cancer tissue microarray ( n = 844). Androstenedione (4AD) levels were evaluated in serum samples ( n = 42) from hormone receptor-positive, postmenopausal breast cancer. Levels of androgens, progesterone, and estradiol were quantified using LC/MS-MS in serum from age- and grade-matched recurrent and nonrecurrent patients ( n = 6) before and after aromatase inhibitor (AI) therapy (>12 months). AR and estrogen receptor (ER) signaling pathway activities were analyzed in two independent AI-treated cohorts., Results: AR protein expression was associated with favorable progression-free survival in the total population (Wilcoxon, P < 0.001). Pretherapy serum samples from breast cancer patients showed decreasing levels of 4AD with age only in the nonrecurrent group ( P < 0.05). LC/MS-MS analysis of an AI-sensitive and AI-resistant cohort demonstrated the ability to detect altered levels of steroids in serum of patients before and after AI therapy. Transcriptional analysis showed an increased ratio of AR:ER signaling pathway activities in patients failing AI therapy ( t test P < 0.05); furthermore, 4AD mediated gene changes associated with acquired AI resistance., Conclusions: This study highlights the importance of examining the therapeutic consequences of the steroid microenvironment and demonstrable receptor activation using indicative gene expression signatures., (©2021 The Authors; Published by the American Association for Cancer Research.)

Context: The gonads are the major source of sex steroids during reproductive ages. The gonadal function declines abruptly in women and gradually in men. The adrenals produce 11-oxygenated androgens (11-oxyandrogens), which start rising during adrenarche. Following menopause, 11-oxyandrogens levels remain similar to reproductive ages., Objective: To compare the circulating 11-oxyandrogen concentrations in men and women across adult ages., Methods: We used mass spectrometry to measure testosterone (T), androstenedione (A4), 11β-hydroxytestosterone (11OHT), 11-ketotestosterone (11KT), 11β-hydroxyandrostenedione (11OHA4), 11-ketoandrostenedione (11KA4), cortisol, and cortisone in morning sera obtained from adults in outpatient setting. We performed double immunofluorescence of 3β-hydroxysteroid dehydrogenase type 2 and cytochrome b5 in adrenal tissue from 19 men, age 23-78 years., Results: We included 590 patients (319 men), aged 18 to 97 years, and 84% white. 11KT and 11KA4 were stable across ages in women, but they declined in men (0.21 and 0.06 ng/dL/year, respectively; P < 0.05). 11OHA4 and 11OHT increased modestly with age in women (0.6 and 0.09 ng/dL/year, respectively; P < 0.01), and both remained stable across ages in men. As body mass index (BMI) increased, 11KA4 decreased in women, and 11KT increased in men, both suggesting higher 17β-hydroxysteroid dehydrogenase activity in obese individuals. A4 and T declined with age and A4 with BMI in both sexes; T declined with BMI in men. Adrenal androgenic enzyme expressions in aging men were similar to those observed in women., Conclusions: In contrast with traditional androgens, the production of 11OHA4 and 11OHT is sustained with aging in both sexes. The bioactive androgen 11KT declines in aging men but not in women., (© Endocrine Society 2020. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

We have previously reported that dehydroepiandrosterone (DHEA) suppresses the activity and mRNA expression of the hepatic gluconeogenic enzyme glucose-6-phosphatase (G6Pase), and hepatic glucose production in db/db mice. Tyrosine phosphorylation levels of Insulin receptor substrate (IRS)1 and IRS2 reportedly differ between the liver and muscle tissue and the effect of DHEA on insulin signaling has not been elucidated. Therefore, we examined DHEA's effect on the liver and muscle tissue of IRS1(-/-) and IRS2(-/-) mice. Eight-week-old male C57BL6, IRS1(-/-), and IRS2(-/-) mice were fed a high-fat diet (HFD), or an HFD containing 0.2% DHEA for 4 weeks. In a separate experiment, 8-week-old male C57BL6 mice were fed an HFD or an HFD containing 0.2% androstenedione for 4 weeks. In an insulin tolerance test, DHEA administration decreased the initial plasma glucose levels in the C57BL6, IRS1(-/-), and IRS2(-/-) mice but did not decrease the ratios to the basal blood glucose level. Although DHEA administration increased Akt phosphorylation in the liver of the C57BL6, IRS1(-/-), and IRS2(-/-) mice, androstenedione administration did not increase Akt phosphorylation in the liver of C57BL6 mice. DHEA administration did not increase Akt and PKCζ phosphorylation in the muscle tissue of C57BL6, IRS1(-/-), or IRS2(-/-) mice. However, androstenedione administration increased Akt and PKCζ phosphorylation in the muscle tissue of C57BL6 mice. These findings suggest that the effect of DHEA on insulin action in the liver is self-mediated by DHEA or DHEA sulfate (DHEA-S) in the presence of IRS1, IRS2, or both., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

We reported previously that bone morphogenetic proteins (BMPs) potently suppress CYP17 expression and androgen production by bovine theca interna cells (TC) in vitro. In this study, real-time PCR was used to analyse gene expression in TC and granulosa cell (GC) layers from developing bovine antral follicles (1-18 mm). Abundance of mRNA transcripts for four BMPs (BMP2, BMP4, BMP6, and BMP7) and associated type I (BMPR1A, BMPR1B, ACVR1 and ACVR1B) and type II (BMPR2, ACVR2A and ACVR2B) receptors showed relatively modest, though significant, changes during follicle development. BMP2 was selectively expressed in GC, while BMP6, BMP7 and betaglycan (TGFBR3) were more abundant in TC. Abundance of betaglycan mRNA (inhibin co-receptor) in TC increased progressively (fivefold; P<0.001) as follicles grew from 1-2 to 9-10 mm. This suggests a shift in thecal responsiveness to GC-derived inhibin, produced in increasing amounts as follicles achieve dominance. This prompted us to investigate whether inhibin can function as a physiological antagonist of BMP action on bovine TC in vitro, in a manner comparable to that for activin signalling. BMP4, BMP6 and BMP7 abolished LH-induced androstenedione secretion and suppressed CYP17 mRNA >200-fold (P<0.001), while co-treatment with inhibin-A reversed the suppressive action of BMP in each case (P<0.001). Results support a physiological role for granulosa-derived inhibin as an antagonist of BMP action on thecal androgen synthesis. A shift in intrafollicular balance between thecal BMP signalling (inhibitory for androgen synthesis) and betaglycan-dependent inhibin signalling (stimulatory for androgen synthesis) accords with the physiological requirement to deliver an adequate supply of aromatase substrate to GC of developing follicles.

We tested whether puberty in golden hamsters is photoperiodically controlled. Hamsters were raised under 14:10 hours Light:Dark (14L) and 1:23 hours Light:Dark (1L) respectively, from birth to 28 days and tested for various parameters. Body weight, Leydig cell (LC) size and testicular testosterone secretion were greater and plasma thyroxin (T4), testicular androstenedione secretion and LC number were lower (P<0.05) in 1L than 14L hamsters. Volumes of testicular components were similar in the two groups. 3beta-hydroxy steroid dehydrogenase immunohistochemistry demonstrated LC progenitors and newly formed adult LC (ALC) in 14L hamsters, which were absent in 1L hamsters; they contained only fetal LC (FLC). Latter findings suggest the presence and absence of postnatally-differerentiated LC in 14L and 1L hamsters, respectively. Androgen results agreed with these findings, because FLC primarily secrete testosterone, and androstenedione is a major androgen secreted by the newly formed ALC. Reduced T4 in 1L hamsters is attributed to the inhibition of thyroid function by the increased duration of melatonin secretion due to non-photostimulatory conditions. The arrest in LC differentiation in 1L hamsters is attributed to low T4 levels. Although the testis size is unaltered under non-photostimulatory conditions, postnatal LC differentiation is inhibited in golden hamsters, and therefore, it is logical to suggest that their puberty is photoperiodically controlled.

Objective: To assess the prevalence of hypertension and use of antihypertensive drug therapy in relation to menopausal status and to delineate perceived associations between androgens and blood pressure in perimenopausal women., Methods: A population-based sample of women aged 50-59 (n = 6893). Women were divided into three groups according to their hormonal status: premenopausal, postmenopausal without hormone therapy, and postmenopausal with hormone therapy., Result: In the premenopausal, postmenopausal without hormone therapy, and postmenopausal with hormone therapy groups, the prevalence of high blood pressure (>/= 140 mmHg systolic or >/= 90 mmHg diastolic) was 43.9, 49.9 and 45.8%, respectively. In women with normal blood pressure, adjusting for age, body mass index and smoking, there were negative associations between serum testosterone and systolic blood pressure in the total sample (P < 0.01) and the postmenopausal without hormone therapy group (P < 0.05). In women using antihypertensive drug therapy with a blood pressure of at least 140/90 mmHg, positive associations were found between serum testosterone and systolic blood pressure in the total series (P < 0.05) and in the postmenopausal without hormone therapy group (P < 0.05)., Conclusion: Abnormal blood pressure is common in middle-aged women regardless of hormonal status. Our findings suggest that testosterone could have a dual influence on blood pressure in perimenopausal women.

Among older mothers, preeclampsia in the first pregnancy was associated with a reduction in maternal breast cancer risk that was significantly more pronounced in women bearing male than female infants. Androgen concentrations in male, preeclamptic pregnancies were consistent with the hypothesis that elevated pregnancy androgens might mediate this apparent modifying effect of fetal gender.

Objective: Neuroendocrine dysfunction in polycystic ovary syndrome (PCOS) was addressed by studying the steroid hormone changes in women with PCOS with either high or normal LH levels leading to inferences regarding the primacy of elevated LH in the pathophysiology of PCOS., Methods: A cross-sectional study was designed in an academic clinical facility involving 234 women with PCOS. Patients were divided into two groups based on an LH/FSH ratio < or >1 and hormonal and metabolic studies were performed in both groups. Factors were determined by binomial logistic regression that predicted group membership of these women., Results: Higher follicular phase estradiol (E2) and androstenedione (A4) levels as well as greater insulin sensitivity were the only factors that predicted the presence of neuroendocrine dysfunction with elevated A4 being necessary for neuroendocrine dysfunction., Conclusions: It was concluded that uncoupling of hypothalamic E2 inhibition by elevated ovarian A4 associated with E2 related sensitization of pituitary LH leads to neuroendocrine dysfunction in PCOS.

Previous studies have demonstrated that ovulatory female goldfish release a variety of sex steroids into the water where they function as a pheromonal blend dominated by C21 steroids that stimulates male hormone release, sperm production and behavior. This study investigated whether male goldfish might also release sex steroids with pheromonal activity. It found that spermiated male goldfish release substantial quantities of androstenedione (AD; about 50 ng/h) together with smaller (10-20 ng/h) quantities of several other related C19 steroids but only very small quantities (<5 ng/h) of C21 steroids. Further, when sexually aroused by females and/or their pheromones, males released even greater quantities of AD (up to 1 microg/h) while C21 steroid release rate changed little. This created a ratio of C19 to C21 steroids of about 50:1 that was dramatically different from that emitted by females (1:7). The male olfactory system was also found to be extremely sensitive to AD, detecting it to near picomolar concentrations. Together with previous studies that have shown water-borne AD to increase male aggressive behavior while suppressing responsiveness to female pheromones, this study establishes AD as a male pheromone in the goldfish. Because ovulating females also release AD but in the presence of C21 steroids, recognition of the male-derived steroid pheromone is presumably mixture dependent.

The objective of this study was to determine if androgens regulate granulosa cell steroidogenesis at physiological doses found in small bovine follicles. Bovine granulosa cells were cultured under serum-free conditions that permit the induction and maintenance of FSH-dependent estradiol secretion. Increasing androstenedione concentrations from 0.1 to 1 or 10 microM significantly increased estradiol accumulation and cytochrome P450 aromatase (P450arom) mRNA abundance. No increase in progesterone accumulation or abundance of mRNA for P450 side-chain cleavage or 3beta-hydroxysteroid dehydrogenase enzymes was observed. The addition of 0.1, 1, or 10 microM progestins or estrogens had no stimulatory effect on P450arom mRNA levels. An analysis of the 5'-untranslated region of P450arom mRNA transcripts indicated that the majority was derived from Cyp19 ovary-specific promoter 2, with some contribution from promoters 1.1 and 1.5. Transcripts from these three promoters were all significantly increased by androstenedione. Testosterone increased promoter 1.1 and 1.5-derived transcripts, but only promoter 2-derived transcripts at the highest dose tested (100 microM). Dihydrotestosterone (DHT) did not affect Cyp19 expression. Collectively, these data show that androgens may exert specific stimulatory effects on P450arom mRNA concentrations in granulosa cells. Interestingly, different androgens had different effects on Cyp19 promoter usage, suggesting differential regulation of aromatase gene expression in the developing follicle., ((c) 2005 Wiley-Liss, Inc.)

In several animal studies, prolactin has been found to be essential for mammary epithelial development, and its administration has been consistently shown to increase the rate of mammary tumours. High levels of steroid hormones have also been suggested to enhance mammary cancer development. The present study investigates the levels of the following hormones in serum and in tissue homogenates in dogs bearing canine mammary tumours: prolactin (PRL), progesterone (P4), dehydroepiandrosterone (DHEA), androstenedione (A4), testosterone (T), 17beta-estradiol (17beta-E2) and estrone sulfate (S04E1). Eighty mammary tumours (40 dysplasias and benign and 40 malignant tumours) from 32 female dogs, and 10 normal mammary glands from eight female dogs without history of mammary tumours, were analysed. Prolactin and steroid hormones in serum and tissue homogenates, were analysed by enzyme immunoassays (EIA) techniques, previously validated for this animal species. Levels of prolactin in tissue homogenates were significantly different between malignant and benign mammary tumours (p<0.01). Serum prolactin concentrations were lower in the control group as compared with the group of dogs with benign tumours and in dogs with malignant tumours (p=0.01). Serum prolactin levels in dogs with benign lesions were not significantly different than those obtained from dogs with malignant tumours. Levels of steroid hormones were significantly higher in malignant tumours compared with the benign tumours and normal mammary glands (p<0.01) both in serum and homogenate determinations. Our results suggest that the canine neoplastic mammary gland could be a source of prolactin. Our hypothesis is that both prolactin and steroid hormones are involved in the growth of canine mammary cancer, and that they might have an autocrine/paracrine role in the maintenance of this disease.

Background: Osteoporosis is a common problem in postmenopausal period. Recent studies have suggested that endogenous and exogenous androgens may influence the bone mineral density in women. There is limited data about the effect of circulating androgens on bone density in postmenopausal women., Aim: The aim of this study was to evaluate the effect of circulating androgens of ovarian and adrenal origin on bone mineral density in postmenopausal women., Materials and Methods: This cross-sectional study included 178 postmenopausal women, who had never been treated with hormonal therapy or calciotropic agents. Serum free testosterone, dehydroepiandrosterone sulfate and androstenedione levels and their relationship with bone mass (dual X-ray absorptiometry) were evaluated., Results: Serum free testosterone and DHEAS levels were correlated positively with bone mineral density at lumbar spine and femoral neck (P < 0.001). However, stepwise linear regression analyses revealed a differential effect of androgens on bone density. Serum free testosterone was among the independent predictor of bone density at lumbar spine (trabecular bone), whereas serum DHEAS level was of bone density at femoral neck (cortical bone)., Conclusion: This study suggests that endogenous androgens are influential on bone density in postmenopausal women. However, regression analyses revealed a differential effect of androgens on different bone types.

Tooth pulp contains steroid receptors and therefore is likely to respond to steroids. Steroids and cytokines together can alter steroid receptor content in many tissues; thus, similar mechanisms may exist in tooth pulp. In this study, reverse-transcription/polymerase chain-reaction was used to screen human pulp for the mRNAs encoding receptors for androgen (AR), estrogens (ERbeta), and hepatocyte growth factor (HGF: c-Met). AR mRNA content was greater in male pulp vs. female pulp in all age groups. In both genders, AR mRNA content diminished with age. In pulp cell cultures, androstenedione, estradiol-17beta, and HGF each stimulated AR mRNA accumulation. Testosterone inhibited, whereas 5alpha-dihydrotestosterone did not affect, AR mRNA content. ERbeta was not hormonally altered in pulp cell cultures. By showing steroid- and cytokine-orchestrated regulation of AR mRNA in vitro, it is possible that age- and/or pathogen-dependent changes in available steroids and cytokines can affect any androgen-responsiveness of pulp.

Although the pathogenesis of polycystic ovarian syndrome (PCOS) is still controversial, a series of investigations has demonstrated an array of neuroendocrine abnormalities as a major component of the syndrome. From a neuroendocrine perspective, patients with PCOS exhibit an accelerated frequency and/or higher amplitude of LH pulses, augmentation of LH secretory burst mass, and a more disorderly LH release. Elevated in vitro LH bioactivity and a preponderance of basic LH isoforms, which correlate positively with elevated serum 17-hydroxyprogesterone, androstenedione, and testosterone concentrations, also characterize adolescents with PCOS. Heightened GnRH drive of gonadotropin secretion and a steroid-permissive milieu appear to jointly promote elevated secretion of basic LH isoforms. Positive feedback is implied, because hypersecretion of highly bioactive LH in PCOS probably contributes to inordinate androgen output. However, the precise nature of feedback disruption remains uncertain. Indeed, recent data suggest that PCOS is marked by anomalies of both feedforward and feedback signaling between GnRH/LH and ovarian androgens. From a single hormone perspective, the individual patterns of LH and androstenedione release are consistently more irregular in patients with PCOS. Bihormonal analysis has disclosed concomitant uncoupling of the pairwise synchrony of LH and testosterone, LH and androstenedione, and testosterone and androstenedione secretion. The foregoing ensemble of findings points to deterioration of both orderly uniglandular and coordinate bihormonal output in PCOS. Additional studies are needed to establish the primary pathophysiologic mechanisms underlying this disorder.

Human glutathione transferase (GST) A1-1 efficiently catalyzes the isomerization of Delta(5)-androstene-3,17-dione (AD) into Delta(4)-androstene-3,17-dione. High activity requires glutathione, but enzymatic catalysis occurs also in the absence of this cofactor. Glutathione alone shows a limited catalytic effect. S-Alkylglutathione derivatives do not promote the reaction, and the pH dependence of the isomerization indicates that the glutathione thiolate serves as a base in the catalytic mechanism. Mutation of the active-site Tyr(9) into Phe significantly decreases the steady-state kinetic parameters, alters their pH dependence, and increases the pK(a) value of the enzyme-bound glutathione thiol. Thus, Tyr(9) promotes the reaction via its phenolic hydroxyl group in protonated form. GST A2-2 has a catalytic efficiency with AD 100-fold lower than the homologous GST A1-1. Another Alpha class enzyme, GST A4-4, is 1000-fold less active than GST A1-1. The Y9F mutant of GST A1-1 is more efficient than GST A2-2 and GST A4-4, both having a glutathione cofactor and an active-site Tyr(9) residue. The active sites of GST A2-2 and GST A1-1 differ by only four amino acid residues, suggesting that proper orientation of AD in relation to the thiolate of glutathione is crucial for high catalytic efficiency in the isomerization reaction. The GST A1-1-catalyzed steroid isomerization provides a complement to the previously described isomerase activity of 3beta-hydroxysteroid dehydrogenase.

The physiological role of 11beta hydroxy-androstenedione (11betaOHA), a primary adrenal steroid, remains unknown. In the present study, we investigated the effect of 11betaOHA on bone metabolism in vitro and in vivo. Administration of 11betaOHA enhanced the clonal growth of marrow osteoprogenitor cells cultured from normal rats. In ovariectomized rats, 11betaOHA restored osteogenesis and increased the bone mineral density at both the metaphyseal and diaphyseal regions of the femur. Bone histomorphometric study of ovariectomized rats demonstrated that the mineral apposition rate of both cortical bone and trabecular bone was increased by treatment with 11betaOHA. In addition, 11betaOHA increased alkaline phosphatase activity in cultured osteoblastic cells (MC3T3-E1 and SaOS-2). The androgenic and anabolic effects of 11betaOHA were respectively estimated to be less than 1/100th and 1/10th-1/100th of those of testosterone, while the estrogenic action of 11betaOHA was also very weak. These findings suggest an influence of 11betaOHA on physiological bone metabolism and indicate that this steroid may be useful for stimulating of bone formation in the treatment of osteoporosis.

Objective: To test the hypothesis that androgens have a direct effect on the function of endometrial epithelial cells., Design: In vitro study., Setting: Academic research center., Patient(s): Endometrial epithelial cells were prepared from biopsy samples obtained from normal fertile women., Interventions: Cells were incubated with androstenedione, testosterone, dihydrotestosterone, and DHEA., Main Outcome Measure(s): Secretion of glycodelin A into the culture fluid was used to assess secretory activity. Uptake of (3)H-thymidine and immunostaining for Ki67 was used to assess cell growth. The specific action of the androgens was confirmed by incubation with an antiandrogen, cyproterone acetate., Result(s): Androstenedione (10(-6) M and 10(-7) M) caused a dose-dependent decrease in glycodelin A secretion, uptake of (3)H-thymidine, and percentage of positive Ki67 cells in cultured human endometrial epithelial cells. Testosterone, dihydrotestosterone, and DHEA had no effect on glycodelin A secretion or (3)H-thymidine uptake. The direct effect of androgens on endometrial function were confirmed by demonstrating the presence of androgen receptors in cultured endometrial epithelial cells and showing that the direct effects of the androgens were not observed when cyproterone acetate was added to the cultures., Conclusion(s): The results suggest that androstenedione can inhibit human endometrial cell growth and secretory activity. Infertility and miscarriage associated with high androgen levels (e.g., that caused by the polycystic ovary syndrome) may be due to an adverse effect of high androgen levels on the endometrium.

Females may favour some offspring over others by differential deposition of yolk hormones. In American kestrels (Falco sparverius), we found that yolks of eggs laid late in the sequence of a clutch had more testosterone (T) and androstenedione (A4) than yolks of first-laid eggs. To investigate the effects of these yolk androgens on nestling 'fitness', we injected both T and A4 into the yolks of first-laid eggs and compared their hatching time, nestling growth and nestling survival with those of first-laid eggs in which we injected vehicle as a control. Compared to controls, injection of T and A4 at a dose intended to increase their levels to those of later-laid eggs delayed hatching and reduced nestling growth and survival rates. Yolk androgen treatment of egg 1 had no effect on survival of siblings hatching from subsequently laid eggs. The adverse actions of yolk androgen treatment in the kestrel are in contrast to the favourable actions of yolk T treatment found previously in canaries (Serinus canaria). Additional studies are necessary in order to determine whether the deposition of yolk androgens is an adaptive form of parental favouritism or an adverse by-product of endocrine processes during egg formation. Despite its adaptive significance, such 'transgenerational' effects of steroid hormones may have helped to evolutionarily shape the hormonal mechanisms regulating reproduction.

There are at least three forms of macrophage colony-stimulating factor (M-CSF), a cytokine that is critical for osteoclast formation; and evidence exists that the membrane-bound form is involved in this process. We wished to test the hypothesis that the expression of the membrane form of M-CSF is modulated by the presence of gender steroids. This was achieved by analyzing M-CSF messenger RNA and protein in the bone-marrow of estrogen- and androgen-replete, and -deficient female rats. We found that the 1.4-kb M-CSF transcript was not detected in sham-operated rats but that the 4.6-kb transcript was expressed in abundance. In contrast, these transcripts were differentially expressed in ovariectomized rats, and this effect was reversed by 17beta-estradiol treatment. Administration of androstenedione to ovariectomized rats, so that androstenedione plasma levels were restored to just below that in sham-operated rats, also suppressed the expression of the 1.4-kb M-CSF transcript. This effect was abrogated by antiandrogen treatment, indicating that this was an androgen-mediated effect. The membrane-bound protein was detected in the bone-marrow of sham-operated rats and was elevated post ovariectomy, whereas ovariectomy had no effect on the soluble isoform. Our data support the hypothesis that the membrane form of M-CSF is modulated by gender hormones and that this isoform is involved in the estrogen- and androgen-mediated effects on the skeleton.

Objective: The aim of this study was to gain understanding of the apparent discrepancy between the moderate restriction of testosterone synthesizing capacity and the nearly complete decay of the androgen-producing enzyme, cytochrome P450c17 (CYP17; steroid 17 alpha-hydroxylase/17,20-lyase), in rat testes during the desensitization phase induced by a single, high-dose gonadotropin (human chorionic gonadotropin, hCG) injection., Design and Methods: Adult male rats received 25 IU hCG i.v., and purified Leydig cells and crude interstitial cell microsomes were prepared 0, 4, 12, 24, 48, 72, 120 and 192 h afterwards. Component CYP17 activities, i.e. simultaneously catalyzed productive androgen formation and abortive 17 alpha-hydroxyprogesterone release, and their ratio (processivity), were compared with CYP17 levels and testosterone secretion rates., Results: Leydig cells isolated 48 h after the artificial hCG surge produce 62% less testosterone than control cells upon stimulation in vitro, though CYP17 levels are reduced by 97%. Its total activity decreases by 87%, resulting in a 4.5-fold rise in the turnover number; the processivity is additionally improved 5-fold over controls. Parallel changes occur in interstitial cell microsomes; a negative linear correlation exists between the ratio of productive over total CYP17 activities and the actual CYP17 concentrations. CYP17 is partly denatured to P420 during hCG action, but other heme proteins (cytochrome b5) remain unchanged. Animal treatment with estradiol results in CYP17 down-regulation without any concomitant effect on enzyme processivity., Conclusion: Improved CYP17 processivity is suggested to be the consequence of (otherwise rate-limiting) improved electron transfer efficiency towards CYP17. It explains the relatively high testosterone secretion during Leydig cell desensitization and is interpreted to be a protective mechanism to confine adverse consequences of enzyme decay.

A low resting metabolic rate (RMR) for a given body composition has been identified as a risk factor for weight gain and obesity, and has also been reported in formerly obese individuals with the genetic predisposition for obesity. The possible role of thyroid hormone in low RMR was studied in a large sample of postobese women. RMR was measured by indirect calorimetry in 28 weight-stable postobese women with a family history of obesity (PO group) and in a control group of 28 nonobese women closely matched for age, fat mass, and fat-free mass. RMR was 8% lower in the PO than in the control group [[symbol: see text]; 95% Cl:5856 (5520, 6214) compared with 6408 kJ/d (6096, 6768 kJ/d), P < 0.02], and the group difference remained unchanged after fat-free mass and fat mass were adjusted for (552 kJ/d, P < 0.015). The PO group had lower plasma free triiodothyronine [2.4 (1.9, 3.0) compared with 3.4 pmol/L (2.9, 3.9 pmol/L), P < 0.01], whereas plasma androstenedione only tended to be lower in the PO than in the control group. Adjustment for differences in androstenedione did not reduce the difference in RMR, whereas adjustment for differences in plasma free triiodothyronine eliminated the group difference (96 kJ/d, P = 0.59). The present study shows that RMR for a given body composition is lower among postobese than among matched never-obese control subjects. Statistically, the lower plasma free triiodothyronine concentrations of the postobese subjects could explain their lower RMRs, but it remains to be established whether these findings are causally related.

Objectives: The relationship between body build, androgen levels and changes in sexual interest after menopause was investigated in 171 postmenopausal women from Vienna, Austria., Methods: All women were interviewed using a structured questionnaire. Body build was determined by employing five absolute body dimensions and four anthropometric indices., Results: Body weight, as well as the amount of subcutaneous centripetal fat (such as in the chest, waist and hip region), were statistically significantly related to the degree of reduced sexual interest. Corpulent and heavy women suffered far more frequently from a severe decrease in sexual interest after menopause. Statistically significant associations between androgen levels and decrease in sexual interest could not be demonstrated., Conclusions: Reduced sexual interest is associated with a kind of body type not corresponding to the culture-specific beauty ideals of our society, first of all evident in women whose menopause occurred relatively early.

An initial group of term (36-41 6/7 weeks), preterm (less than 36 weeks), and post-term (42 or more weeks) placentae were collected from women at delivery to determine the placental levels of important steroids and steroidogenic enzymes involved in the oestrogen synthesis pathway as a function of gestational age. A second group of placentae were obtained from women delivering at term before and after the onset of labour. Placentae were evaluated individually for cytosolic steroid hormone levels and microsomal steroidogenic enzyme activities. Oestradiol (E2), oestrone (E1), progesterone (P), and delta-4-androstenedione (A) were measured by radioimmunoassay in placental cytosols. Aromatase (AR), sulphatase (S), and 3 beta-hydroxysteroid dehydrogenase/isomerase (3 beta HSD) activities were assayed in placental microsomes. Cytosolic concentrations of E1, E2, P, and A did not differ with respect to gestational age. Correspondingly, the microsomal enzyme activities of 3 beta HSD, S, and AR did not vary as a function of gestational age. However, when patients at term who were in labour prior to delivery were compared to those who were not, the placental cytosolic level of E1 was found to be threefold higher in the non-labouring group (4572 versus 1427 pg/mg cytosolic protein, P < 0.025). Additionally, microsomal aromatase activity was also significantly higher in the non-labouring patients (46 versus 19 pM/min/mg protein, P < 0.025), while the E2 to P ratio in the labouring patients was twice that of the non-labouring group, a difference which was significant at the P < 0.025 level (Wilcoxon rank sum test). These data suggest that at term, prior to labour, the placental production of E1 by AR is high, and that AR activity and E1 levels fall significantly after the onset of labour. Also, the placental cytosolic concentration of the more active oestrogen, E2, demonstrates stable to rising levels with a significant increase in E2/P after the onset of labour. We theorize that in the term pregnancy prior to labour, E1 may represent a large but relatively inactive intracellular oestrogen pool which is maintained by high AR activity, and may function to protect the pregnant local uterine environment from the more oxytocic effects of E2.

An experiment was carried out using 320 adult Merino ewes to examine the effects of immunization against an androstenedione human serum albumin conjugate (Fecundin) on ovulation rate, fertilization rate and embryo viability at days 2, 9 and 13-14 after fertilization. The ovulation rate of immunized ewes (2.19 +/- 0.06) was greater (P < 0.001) than that of control ewes (1.43 +/- 0.04). The recovery rate of embryos or of unfertilized oocytes on day 2 was reduced in immunized ewes, but fertilization rate of recovered oocytes was unaffected by immunization. The mean number of normal embryos per ewe pregnant (prolificacy) was higher and the proportion of ewes pregnant (fertility) was lower in immunized than in the control ewes. The distribution of the number of cells per embryo showed no differences in developmental age over the period of sampling, the majority of embryos at this time being at the two- to four-cell stage of development. At day 9 of pregnancy, blastocyst recovery rates were lower in immunized than in control ewes. About 90% of blastocysts recovered were developing normally in control ewes compared with 64% in immunized ewes. The majority of blastocysts recovered on day 9 had hatched from the zona pellucida prior to recovery (mean values were 94.2% and 87.8% for control and immunized groups, respectively). In control ewes single blastocysts were larger than twin blastocysts, but for the immunized ewes this difference was not significant. Both single blastocysts (P < 0.01) and twin blastocysts (P < 0.05) from immunized ewes were smaller than those from control ewes.(ABSTRACT TRUNCATED AT 250 WORDS)

In 20 women with proven fertility, one menstrual cycle was monitored by ovarian ultrasonography, laparoscopy and estimation of 17 beta-estradiol (E2), estrone (E1), progesterone, testosterone, androstenedione (Adion), dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate (DHEAS), and sex hormone binding globulin (SHBG) in serum and peritoneal fluid. Three groups were studied, in which the peritoneal fluid was collected within 1, 3 and 5 days after ovulation. E1, E2 and progesterone levels in peritoneal fluid were highest shortly after ovulation and decreased with time. Testosterone and Adion in peritoneal fluid showed no changes, but peritoneal fluid levels were always higher than serum levels. No differences were found between the peritoneal fluid and serum levels of DHEA and DHEAS. SHBG in serum was always higher than in peritoneal fluid. The results are compared with reported steroid levels in follicular fluid from the literature and factors complicating the interpretation of steroid levels in peritoneal fluid are discussed.

Two experiments were undertaken to determine the hormonal response of Merino ewes to immunization against androstenedione (Fecundin). In Exp. 1 peripheral concentrations of LH, FSH and progesterone were monitored in spontaneously cycling ewes (20 immunized and 21 controls). In Exp. 2 (10 immunized and 10 controls) the same hormones were measured in ewes before and after prostaglandin (PG)-induced luteolysis and, in addition, the pattern of pulsatile LH secretion was determined during the luteal (PG + 12 days), early follicular (PG + 24 h) and late follicular (PG + 40 h) phase of the oestrous cycle. Ovulation rates were measured in both experiments. The results of these experiments indicate that androstenedione-immune animals have elevated ovulation rates (0.6-0.7 greater than control animals; P less than 0.05) associated with elevated plasma concentrations of LH and progesterone. The magnitude of the increase in plasma progesterone was correlated with androstenedione antibody titre (r = 0.6, P less than 0.001). LH pulse frequency of androstenedione-immune ewes tended to be higher at all stages of the oestrous cycle, but this difference was only significant (P less than 0.05) during the luteal phase. Mean plasma concentrations of FSH did not differ significantly between immunized and control ewes at any stage of the cycle. Analysis of periodic fluctuations in FSH during the luteal phase revealed that androstenedione-immune animals had a similar number of fluctuations of a similar amplitude to those of control animals, but the nadir of these fluctuations was lower (P less than 0.05) in immunized animals. A significant (P less than 0.05) negative correlation existed between androstenedione antibody titre and the interval between FSH peaks (r = -0.49) and androstenedione antibody titre and FSH nadir concentrations (r = -0.46). It is concluded that plasma FSH concentrations are not a determinant of ovulation rate in androstenedione-immune ewes and that increased LH concentrations, or perturbation of normal intraovarian mechanisms, may be responsible for the increase in ovulation rate observed in ewes immunized against androstenedione.

Unlabelled: We studied the effects of chronic renal failure on the pituitary-cortisol axis and adrenal androgen function in 26 patients (16 male and 10 female), aged 6.5 to 22.5 years (mean 14.5). Ten patients were prepubertal, 8 pubertal, and 8 post-pubertal. All of them were on chronic hemodialysis. Pubic hair development was delayed in 56% of the patients. Serum cortisol was increased in 15 out of the 26 patients. Serum delta 4-androstenedione was high in 11 out of 15 patients in Tanner's stage I or II and in 1 out of 11 patients in Tanner's stage III, IV or V (p less than 0.01). Serum cortisol was elevated in 10 out of 12 patients with high serum delta 4-androstenedione and in only 5 out of 14 with normal delta 4-androstenedione (p less than 0.02). Serum dehydroepiandrosterone sulphate was normal in 22 patients and elevated in 4 males. There was a significant inverse correlation between bone age and serum cortisol (r: -0.59; p less than 0.005) and a significant positive correlation between bone age and serum dehydroepiandrosterone sulphate (r: 0.45 p less than 0.01). Serum ACTH was normal. A reduction by 50% in cortisol and 78% in dehydroepiandrosterone sulphate was found after dexamethasone suppression, but delta 4-androstenedione did not suppress after dexamethasone. After ACTH stimulation test cortisol increased by 50% and delta 4-androstenedione by 80%., Conclusions: The increased levels of cortisol and delta 4-androstenedione with partial resistance to dexamethasone suggest that these patients have a hypothalamic-pituitary dysfunction similar to that found in Cushing's disease or in chronic stress. The difference in the responses of delta 4-androstenedione and dehydroepiandrosterone sulphate observed is consistent with the existence of different mechanisms of control for these two steroids.

Active immunization of sheep against androstenedione results in an increase in ovulation rate that is associated with increased plasma levels of LH and progesterone, but not FSH. Although immunized ewes have more activated follicles the secretion rate of oestradiol is not increased. An experiment was conducted to examine the effect of androstenedione immunity on the ovarian secretion and peripheral plasma concentrations of inhibin. Merino ewes in which the left ovary had been autotransplanted to a site in the neck were divided into control (n = 5) and androstenedione-immune (n = 6) groups. Ovarian and jugular venous blood was collected every 10 min at two stages of the follicular phase, 21-27 h and 38-42 h after a luteolytic dose of an analogue of prostaglandin F2 alpha (PG), and every 15 min for 6 h on day 10 of the subsequent luteal phase. The ewes were monitored regularly for luteal function by measurement of the concentration of progesterone and preovulatory LH surges. The concentration of inhibin in jugular and ovarian venous plasma was determined by radioimmunoassay and ovarian secretion rates and peripheral concentrations are expressed as pg of 1-26 peptide fragment of the alpha chain. The ovarian secretion rate of inhibin tended to be greater in androstenedione-immune ewes at all stages of the oestrous cycle measured, with this difference being statistically significant (P less than 0.05) during the luteal phase (100 +/- 40 and 260 +/- 80 (S.E.M.) pg/min for control and immune groups respectively). The pattern of ovarian inhibin secretion exhibited pulsatile-like fluctuations which were not associated with LH pulses. Peripheral concentrations of inhibin were generally higher in immunized than in control ewes with this difference being significant (P less than 0.01) from day 4 to 14 of the luteal phase (59 +/- 5 and 110 +/- 7 ng/l for control and immune respectively). The ovarian secretion rate of immunoactive inhibin was greater (P less than 0.01) during the follicular phase than during the luteal phase in both groups of ewes, and peripheral concentrations of inhibin increased (P less than 0.001) following injection of PG in ewes from both treatment groups. We concluded that androstenedione immunity results in an increase in ovarian inhibin secretion, an effect that can probably be attributed to the greater number of large oestrogenic follicles present in the ovaries of these ewes.(ABSTRACT TRUNCATED AT 400 WORDS)

There is little information about the stimulation of estrogen secretion from theca cells of the domestic hen by follicle-stimulating hormone (FSH), and the mechanism of action of FSH through calcium has not been considered previously. The theca interna and externa cells from the third (F3) and fourth (F4) largest ovarian follicles of hens were separated, dispersed, and incubated in M199 with FSH (0.5 micrograms ml-1) or A23187 (0.1 or 1 microgram ml-1) with or without ethylene glycol bis(beta-aminoethyl ether)N,N'-tetraacetic acid (EGTA) (2 mM) in calcium-replete or in calcium-free M199. Alternatively, androstenedione (10(-6) M) was added to the cells as aromatizable substrate, with or without FSH and/or EGTA. Estradiol and estrone secreted into the media during a 4-hr incubation period were measured by RIA. FSH stimulated estradiol and estrone secretion from all the cell preparations. The effect of FSH was abolished by the addition of EGTA or in calcium-free medium. A23187 stimulated estradiol and estrone secretion by the same extent as FSH, but did not do so in calcium-free medium. Androstenedione enhanced estradiol and estrone secretion, but estrogen secretion was further raised by the simultaneous addition of FSH. This action of FSH on aromatization was also abolished by EGTA. This evidence supports the hypothesis that calcium, possibly of extracellular origin, is an important mediator in the stimulation of aromatase systems in thecal cells of chickens.

Groups of heifer calves received a primary immunization against androstenedione (Group A; N = 11) or oestradiol-17 beta (Group E; N = 10) at 3 months of age and booster injections on 5 occasions at 2- to 3-month intervals. Controls (Group C, N = 11) were immunized against human serum albumin alone using the same protocol. Immunity was achieved against both steroids as judged by the secondary antisteroid antibody titres in Group A (1126 +/- 261; reciprocal of titre) and Group E (10,357 +/- 4067) heifers. In Groups A and E there was a general decline in the respective peak antibody titres after successive booster injections. From 3 to 9 months of age mean plasma concentrations of LH were higher (P less than 0.05) in Group E heifers (0.89 +/- 0.08 ng/ml) than in Group C (0.46 +/- 0.03 ng/ml) and Group A (0.59 +/- 0.05 ng/ml) heifers which did not differ from one another. There were no differences between groups in plasma FSH concentrations. At 10 months of age the LH response to exogenous LHRH was of higher (P less than 0.05) amplitude for heifers in Group E (2.59 +/- 0.56 ng/ml) than for those in Groups C (0.61 +/- 0.07 ng/ml) and A (1.04 +/- 0.22 ng/ml). Elevated plasma progesterone concentrations at 5 months of age were shown by 2 heifers in Group C, 10 in Group A, and 6 in Group E. From 8 to 14 months of age a consistently higher proportion of Group A heifers exhibited elevated progesterone compared with Group C and Group E heifers. After ovarian synchronization and booster injection at 15 months of age a corpus luteum was present in 2 heifers in Group C, 7 in Group A and none in Group E. The ovaries of Group A heifers were different from those of Groups C and E and were characterized by greater numbers of 2-4 mm follicles. It is concluded that active immunization against gonadal steroids influences both LH secretion and ovarian function in prepubertal heifers. Early increases in ovarian activity in androstenedione-immunized heifers are maintained after puberty and may therefore confer some lifetime reproductive advantages.

The incidence of oestrus (6/46) and ovulation (14/46) in ewes given antisera to androstenedione, oestrone, oestradiol and testosterone either separately or as a mixture of these sera at the time of treatment with progestagen sponges alone or progestagen sponges followed by LH-RH was similar to that of control ewes (2/13 and 6/13 respectively). The number of corpora lutea (CL) recorded for those ewes that did ovulate was, however, greater in the antiserum-treated ewes (22 CL/14 ewes) than in the controls (6 CL/6 ewes) at the first ovulation after sponge withdrawal. This superiority persisted to the second ovulation (53 CL/42 treated ewes compared to 13 CL/13 controls). The results for groups treated with antisera did not differ amongst themselves.

Both the adrenal and the ovary contain the biosynthetic pathways necessary for androgen synthesis and secretion. The fetal ovary is not very active but the fetal adrenal is an important source of DHAS. However the secretion of DHAS declines markedly after birth and until puberty there is little androgen secretion by either the adrenal or the ovary. Post-pubertally, the adrenal secretes DHAS, DHA, delta 4-A and T from the reticularis and probably the fasciculata. This secretion is under ACTH control, at least in part, but apparently also under control of another pituitary polypeptide tentatively called 'adrenal androgen secretory hormone'. THe adrenal secretion rates are in the range of 7-14 mg/day for DHAS, 3-4 mg/day for DHA, 1-1.5 mg/day for delta 4-A and 50 micrograms/day for T. Androgen secretion from the ovary arises in part from the theca cells of the follicle, the corpus luteum and the stromal cells, under LH control, and will vary somewhat during the normal menstrual cycle. The ovarian secretion rate in the follicular phase is 1-2 mg/day for DHA, 1-1.5 mg/day for delta 4-A and about 50 micrograms/day for T. In the peri-ovulatory period the secretion rate of delta 4-A can rise to 3-3.5 mg/day but there appears to be little change in the secretion of DHA and T. The normal ovary does not secrete significant amounts of DHAS. In about 50% of post-menopausal women the ovaries continue to secrete some T but little delta 4-A or DHA.

In ewes immunized against androstenedione, ovulation rate was increased (P less than 0.001) although the number of live lambs/nursing ewe (1.37), but not the sex ratio (10 females:1 male), was similar to that of controls (1.33).

Heifers were treated with saline (Group I, N = 10), keyhole limpet haemocyanin (KLH; Group II, N = 10), androstenedione-KLH antigen (Group III, N = 14), or oestradiol-17 beta-KLH antigen (Group IV, N = 14). Booster injections were given to produce binding of greater than 10% at dilutions of 1:100 to 1:1000 (50% binding = 14 X 4 pg androstenedione and 9 X 5 pg oestradiol). The heifers were mated and killed at approximately 46 days of gestation to establish ovulation rates, calf numbers, blood hormone relationships and ovarian morphology. Ovulation rate in animals immunized against androstenedione (Group III) was significantly greater than in the other groups; 4 of the animals had double ovulations and 3 had twins. No significant differences were found between Groups I, II and IV in relation to ovulation or pregnancy rate and animals immunized against oestradiol-17 beta continued to cycle and become pregnant. Systemic progesterone, androstenedione and oestrogen levels were generally increased in Groups III and IV but the differences were not significant. No differences were detected between treatment groups in relation to CL weights, ovarian weights, follicle sizes or numbers. No binding of [3H]androstenedione or [3H]oestradiol-17 beta was detected in allantoic or amniotic fluids or fetal serum. Maternal antibody binding was correlated with binding of [3H]androstenedione and [3H]oestradiol in the follicular fluid sampled from the two largest follicles in the ovaries of each animal (Group III, r = 0 X 59; Group IV, r = 0 X 60; P less than 0 X 05).(ABSTRACT TRUNCATED AT 250 WORDS)

The incidence of routine ovariectomy approximates 20% to 30% of all women at hysterectomy. The propriety of this practice is evaluated from three perspectives: the review of the longevity of ovarian hormonal function throughout life, the review of the low risk of subsequent disease in the retained ovary, and the review of epidemiologic considerations. Because oophorectomy and the loss of its steroid contribution has such a profound influence on many body functions, with the most devastating relation to osteoporosis, and because there are no meaningful data in the literature to support the value of routine oophorectomy, removal of ovaries should only be performed when the ovaries are diseased.

Normal sexual development is the consequence of three sequential interrelated processes: establishment of genetic, gonadal and somatic sex. It is the terminal phase of sexual differentiation--the translation of gonadal into somatic sex, which is governed by the presence or absence of both testosterone and Müllerian-inhibiting hormone and of dihydrotestosterone, which is formed in its respective target tissues. Thus, despite a testis, somatic male sexual differentiation will proceed to a normal male phenotype only if all three hormones are synthesized and act during a critical period of uterine development. Many clinically distinct syndromes are the results of abnormalities in the synthesis or action of the above-mentioned hormones; these syndromes are described in detail. In contrast to male somatic differentiation, female somatic development is independent of these hormones.

The effects of progesterone priming and active immunization against androstenedione on ovulation, oestrus ad premature luteal regression were observed in seasonally anovular Merino ewes which had been induced to ovulate by the introduction of rams. Ovulation was induced in 27 out of 36 ewes, and the response was not affected by either immunization or progesterone priming. The ovulation rate (mean number of ovulations per ewe ovulating) was higher in ewes immune to androstenedione, but there was no significant effect of this treatment on oestrus or luteal maintenance. Priming with progesterone prior to the introduction of rams prevented premature regression of the corpora lutea but had no effect on the other measures of reproductive function. If active immunization were to be applied on a commercial scale, it would not interfere with the practice of mating ewes during the anoestrous season. The increase in ovulation rate in these ewes, with no increase in the proportion of ewes ovulating, indicates that the mechanism for the control of ovulation is separate from that controlling ovulation rate. The mechanism by which progesterone pretreatment prevents premature luteal regression awaits further investigation.