The secretion of androgens and estrogens by normal and abnormal testes was compared by determining the concentrations of dehydroepiandrosterone (DHEA), <em>androstenedione</em> (Delta(<em>4</em>)A), testosterone (T), estrone (E(1)), and 17beta-estradiol (E(2)) in peripheral and spermatic venous plasma samples from 1<em>4</em> normal men and 5 men with unilateral testicular atrophy. Four normal men and one patient with unilateral atrophy of the testis were given human chorionic gonadotropin (HCG) before surgery. Plasma estrogens were determined by radioimmunoassay; plasma androgens were measured by the double-isotope dilution derivative technique. Peripheral concentrations of these steroids before and after HCG were similar in both the normal men and the patients with unilateral testicular atrophy. In normal men, the mean +/-SE spermatic venous concentrations were DHEA, 73.1+/-11.7 ng/ml; Delta(<em>4</em>)A, 30.7+/-7.9 ng/ml; T, 751+/-11<em>4</em> ng/ml; E(1), 306+/-55 pg/ml; and E(2), 1298+/-216 pg/ml. Three of four subjects with unilateral testicular atrophy had greatly diminished spermatic venous levels of androgens and estrogens. HCG treatment increased the testicular secretion of DHEA and T fivefold, Delta(<em>4</em>)A threefold, E(1) sixfold, and E(2) eightfold in normal men. In the single subject with an atrophic testis who received HCG, the spermatic venous concentrations of androgens and estrogens were much less than in normal men similarly treated. We conclude that: (a) E(1) is secreted by the human testis, but testicular secretion of E(1) accounts for less than 5% of E(1) production in normal men; (b) HCG stimulation produces increases in spermatic venous estrogens equal to or greater than the changes in androgens, including testosterone; and (c) strikingly decreased secretion of androgen and estrogen by unilateral atrophic human tests cannot be appreciated by analyses of peripheral steroid concentrations.

Studies within the Arab population in Israel revealed 25 pseudohermaphrodites due to 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD) deficiency. Twenty-three individuals, presently living in the Gaza strip, belong to a very large inbred kinship which extends over 8 generations. All affected subjects (<em>4</em>6, XY) were born with mild to moderate degrees of ambiguity of an apparently normal-looking female genitalia and therefore were reared as girls. In childhood, genital abnormalities consisted of a clitoral-like phallus surrounded by a chordee, non-fused labial-scrotal folds and a urogenital sinus. The testes were in the inguinal canals, or rarely, in the labial-scrotal folds. Wolffian structures were normally differentiated while Mullerian structures were absent. At puberty, subjects developed a male body habitus with abundant body hair and beard. Gynecomastia was absent. The phallus and testes enlarged to adult proportions while the prostate remained small. Together with the physical change from girls to boys they developed a male identity having erections and ejaculations, which in 7 cases led to the spontaneous adoption of a male gender role. In adults the hormonal abnormalities consisted of greatly elevated delta <em>4</em>-<em>androstenedione</em> (delta <em>4</em>) (350-1267 ng/dl) associated with subnormal testosterone (T) levels (0.9-3.1 ng/ml). Dihydrotestosterone (DHT) levels, with the exception of 1 patient, were relatively low in all cases (27-35 ng/dl). Children had low levels of delta <em>4</em>, T and DHT, which were normal for age. Although from puberty on there was a significant rise of the 3 androgens, delta <em>4</em> always remained extremely elevated and T and DHT relatively low when compared to normal controls. Dexamethasone failed to suppress the androgen pattern while HCG augmented the defect, making the diagnosis possible in 2 prepubertal children. Dehydroepiandrosterone (DHEA) and 17-hydroxyprogesterone (17-OHP) levels were normal or moderately elevated. Estradiol (E2) levels were normal in children and all but 2 adults, who had high levels. LH and FSH levels were very high after puberty, but normal before. However, there was an overresponse to LHRH in all age groups. The contrast between the lack of intrauterine virilization of the external genitalia in fetuses with 17 beta-HSD deficiency versus the marked masculinization that occurs after puberty still remains a puzzling phenomenon. It is conceivable that the postpubertal development of a male phenotype with change of gender identity and role occurs due to the joint effect of delta <em>4</em>, T and DHT, even though secreted in inadequate proportions. Thus masculinization in these individuals is a slow process requiring a longer period of time than that of normal puberty to be completed.

The theca cells (TC) first become identifiable in preantral follicles after the granulosa cells (GC) begin to divide. It remains unknown when the TC first respond to LH and acquire the capacity to produce androgens. The signal initiating TC differentiation is also unknown since pre-theca cells do not contain LH receptors. Since the first wave of follicle development in the rat occurs postnatally, we correlated the function of dispersed ovarian cells from <em>4</em>-, 5-, 6-, 7-, and 10-day-old rats with the morphological differentiation of TC. The largest follicles in ovaries from <em>4</em>-day-old rats were primary follicles without associated TC. These cells were unable to produce cAMP or steroids in vitro in response to hCG. At 5 days, the first theca were associated with follicles containing 2-3 layers of GC. These cells were responsive to hCG, producing cAMP, progesterone, <em>androstenedione</em>, and androsterone. Responses to hCG increased progressively through 10 days of age. Cholesterol side-chain cleavage (P<em>4</em>50scc), 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD), and 17 alpha-hydroxylase/C17-20 lyase (P<em>4</em>50(17 alpha)) enzymes were localized exclusively to the theca interna. Messenger RNAs for LH receptor, P<em>4</em>50scc, 3 beta-HSD, and P<em>4</em>50(17 alpha) were expressed prior to the time the TC become responsive to LH or morphologically differentiated. To determine the source of the signal regulating TC differentiation, dispersed cells from <em>4</em>-day-old rat ovaries that were unresponsive to LH were treated with preantral follicle-conditioned medium containing thecal differentiating factor (TDF) activity. The TDF activity stimulated androgen production and expression of LH receptor, P<em>4</em>50scc, 3 beta-HSD, and P<em>4</em>50(17 alpha) mRNAs. These data demonstrate that a paracrine signal from the preantral follicle can initiate TC differentiation prior to expression of LH receptors. TC become responsive to LH and capable of producing androgens coincident with morphological differentiation.

Ovarian granulosa cells are the primary site of estrogen and progesterone synthesis and play an essential role in the maturation of the developing ovum. Freshly isolated granulosa cells are often used to study the regulation of steroid and protein biosynthesis, but the small number of cells available for these cultures has proven inadequate for many detailed gene regulatory studies. The goal of this study was to develop human granulosa (HG) cell lines that maintain differentiated function. The E6 and E7 open reading frames of high risk strains of human papillomavirus have been used to produce immortalized cell lines. Primary cultures of human luteinized granulosa cells were infected with defective retroviruses containing the E6 and E7 regions of human papillomavirus 16 and with the neomycin phosphotransferase gene to confer G<em>4</em>18 resistance. Three of eight clones that were isolated after selection in medium containing G<em>4</em>18 were found to produce progesterone following treatment with forskolin or dibutyryl cAMP for <em>4</em>8 h. Forskolin caused these cells to retract in the characteristic rounding response, as described in primary HG cultures. One clone, HGL5, was used for a detailed characterization of differentiated function. HGL5 cells retained the ability to increase progesterone production and convert exogenously added <em>androstenedione</em> to estradiol in response to agonists of the protein kinase-A pathway (forskolin and dibutyryl cAMP), but were not responsive to FSH or LH treatment. A key enzyme in the production of estradiol, cytochrome P<em>4</em>50 aromatase, has proven difficult to maintain in long term cultures of granulosa cells. For that reason, we examined the expression of aromatase in the transformed HGL5 clone by monitoring mRNA levels. Aromatase mRNA increased by <em>4</em>- to 5-fold after forskolin treatment, as determined by Northern analysis. This human granulosa cell culture line maintains many of the functions of normal cells and should provide an important model to study the molecular events controlling granulosa cell differentiation and function.

Progesterone production by the corpus luteum (CL) is essential for preparation of the endometrium for implantation and for the maintenance of gestation. Progesterone modulates its own production and opposes functional luteal regression induced by exogenous agents, such as prostaglandin F(2alpha). In the present study, we evaluated whether progesterone is also capable of interfering with the process of structural luteal regression, which is characterized by a decrease in weight and size of the gland because of programmed cell death (i.e., apoptosis). We have found that a low number of luteal cells undergo apoptosis throughout gestation. On the day of parturition, but following the initial decline in endogenous progesterone production, a small increase in the number of luteal cells undergoing cell death was observed. This increase in apoptotic cells continued postpartum, reaching dramatic levels by Day <em>4</em> postpartum, and was accompanied by a marked decrease in average luteal weight. We have established that the exogenous administration of progesterone significantly reduces the decline in luteal weight observed during structural luteal regression postpartum. This effect was associated with a decrease in the number of cells undergoing apoptosis and with enhanced circulating levels of <em>androstenedione</em>. Furthermore, in vivo administration of progesterone delayed the occurrence of DNA fragmentation in postpartum CL incubated in serum-free conditions. Finally, we have shown that neither the CL of gestation nor the newly formed CL after postpartum ovulation express the classic progesterone-receptor mRNA. In summary, the present results support a protective action of progesterone on the function and survival of the CL through inhibition of apoptosis and stimulation of <em>androstenedione</em> production. Furthermore, this effect is carried out in the absence of classic progesterone receptors.

The endocrine effects of replacement doses of hydrocortisone in postmenopausal women with advanced breast cancer were compared with the same doses of hydrocortisone plus aminoglutethimide. Fifteen patients received aminoglutethimide (AG) 250 mg three times a day plus hydrocortisone (HC) 20 mg twice a day for 2 weeks, then AG was increased to 250 mg four times a day. Another 13 patients received HC alone for 2 weeks, then AG was added. HC alone significantly suppressed oestrone (75% of baseline) and oestradiol (50% of baseline). Addition of AG to these patients produced further oestrone suppression (50% of baseline) significantly greater than HC alone. HC alone suppressed dehydroepiandrosterone sulphate as much as AG + HC. delta <em>4</em>-<em>androstenedione</em> (delta <em>4</em>A) and dehydroepiandrosterone (DHA) were suppressed by HC alone. Addition of AG produced a rise of delta <em>4</em>A to basal levels. These results show that 3-beta-ol de hydrogenase is not induced by AG. AG plus HC together from day 1 produced significantly greater oestrone suppression (50% of baseline) than HC alone. Because high-dose steroids may induce aromatase and replacement doses produced marked peripheral endocrine effects, the use of replacement hydrocortisone should be reassessed in advanced breast cancer.

Amniotic fluid androgen and estrogen levels associated with <em>4</em>8 male and 72 female fetuses between 1<em>4</em> and 20 weeks of gestation were measured. Amniotic fluid testosterone levels were significantly higher (P less than 0.001) in the male (22<em>4</em> +/- 11 pg/ml) than the female fetuses (39 +/- 2 pg/ml) with no overlap of values. Amniotic fluid <em>androstenedione</em> concentrations were also significantly higher (P less than 0.001) with male (102<em>4</em> +/- 53 pg/ml) than female fetuses (668 +/- 39 pg/ml), but there was overlap. There was no difference between anmiotic fluid dehydroepiandrosterone levels for the two sexes. Estrone concentrations were slightly but not significantly higher with the male (353 +/- 33 pg/ml) than with female fetuses (331 +/- 28 pg/ml), while estradiol concentrations were significantly higher (P =0.002) with the female (96 +/- 8 pg/ml) than male (6<em>4</em> +/- <em>4</em> pg/ml) fetuses. It is interpreted that the higher amniotic fluid testosterone and <em>androstenedione</em> levels for the male fetuses reflect fetal testicular secretion. The significantly higher estradiol concentrations for the female fetuses may reflect early ovarian secretion.

A cross-sectional study of 232 healthy children, with about equal numbers of boys and girls and blacks and whites, aged <em>4</em> to 16 yr, was conducted to investigate the racial differences in bone mineral. Bone mineral content (BMC) by dual x-ray absorptiometry was found to be similar between blacks and whites at the spine after controlling for age and Tanner stage. However, total body BMC was higher in blacks, compared with whites of the same age and Tanner stage. Height and weight alone reduced the racial difference in BMC from 152 g to 66 g in girls and from 163 g to 105 g in boys, in whom the difference was further reduced to 66 g after accounting for lean and fat body mass and subscapular skinfold. The only significant sex hormone was <em>androstenedione</em>, which explained another <em>4</em>-5 g of the racial difference in total body BMC for both boys and girls. Among the biochemical variables, only 25OH vitamin D reduced the residual racial difference in total body BMC to 39 g in girls, whereas serum PTH, urine free deoxypyridinoline ratio, and 1,25(OH)(2) vitamin D reduced the residual difference to 25 g in boys. The residual racial differences in bone mass were not statistically significant.

Plasma levels of dehydroepiandrosterone sulfate (DHEA-S), testosterone, dihydrotestosterone (DHT) <em>androstenedione</em>, sex hormone-binding globulin (SHBG), lipoproteins, apolipoproteins and high density lipoprotein (HDL) subfraction were measured in 32 men aged 26-<em>4</em>0 years after myocardial infarction (MI) suffered at least 3-<em>4</em> months prior to the study, who were normocholesterolemic and had angiographically demonstrated coronary occlusion. The control group consisted of 76 healthy men aged 25-<em>4</em>0 years. Blood samples were obtained in the morning from fasting subjects. A significant decrease in plasma DHEA-S and DHT levels were found in MI patients. Also, a significant decrease in HDL-cholesterol, HDL2-cholesterol (HDL2-C) and apolipoprotein A-I, an increase in apolipoprotein B and LDL-cholesterol (LDL-C) levels were observed in those patients as compared with healthy men. However, there were no differences in testosterone, <em>androstenedione</em> and SHBG concentrations between the groups. Significant correlations between testosterone and HDL2-C (r = 0.<em>4</em>6, P less than 0.01), as well as between DHEA-S and HDL3-C (r = 0.39, P less than 0.05) levels in MI patients were observed. These results suggest that decreased levels of plasma DHEA-S and DHT may promote the development of coronary atherosclerosis in men.

The aim was to investigate potential interactions between FSH and intraovarian growth factors in modulating secretion of inhibin A (inh A), activin A (act A), follistatin (FS), estradiol (E2), and progesterone (P<em>4</em>) by bovine granulosa cells cultured under conditions in which a nonluteinized FSH-responsive phenotype is maintained. Cells from <em>4</em>- to 6-mm follicles were cultured in serum-free medium containing insulin (10 ng/ml) and <em>androstenedione</em> (10(-7) M), and effects of ovine FSH (0.037-3 ng/ml) were tested alone and in combination with insulin-like growth factors (IGF) (LR3 IGF-I analogue; 2-50 ng/ml) and epidermal growth factor (EGF; 0.1-10 ng/ml). Medium was changed every <em>4</em>8 h and cultures ended after 1<em>4</em><em>4</em> h, when cell number was determined. Between <em>4</em>8-96 h and 96-1<em>4</em><em>4</em> h, FSH promoted (P < 0.0001) increases in output of inh A (6-fold), act A (15-fold), FS (6-fold), and E2 (18-fold), with maximal responses (in parentheses) elicited by 0.33 ng/ml FSH during the final period. Higher FSH doses (1 and 3 ng/ml) gave reduced responses for each of the above hormones, whereas P(<em>4</em>) output was maximal (3-fold) at these doses. FSH promoted a slight increase in cell number ( approximately 1.7-fold; P < 0.001). LR3 IGF-I alone markedly increased (P < 0.0001) output of inh A (8-fold), act A (<em>4</em>1-fold), FS (12-fold), and E2 (18-fold); this was accompanied by modest increases (P < 0.01) in P<em>4</em> output ( approximately 2.5-fold) and cell number ( approximately 2-fold). Whereas FSH enhanced inh A, act A, FS, and E2 secretion evoked by lower doses of LR3 IGF-I, it suppressed (P < 0.001) the response to the highest dose. EGF alone promoted a 1.7-fold increase in cell number (P < 0.001) without affecting hormone release; however, it abolished (P < 0.001) FSH-induced secretion of inh A, act A, FS, and E2. Both FSH alone and LR3 IGF-I alone dose-dependently increased the act A:FS ratio ( approximately 3-fold; P < 0.005) and act A:inh A ratio (3-fold to 6-fold; P < 0.001), suggesting that both factors selectively raise activin "tone" and that this could be a key requirement for FSH and IGF-induction of follicular E2 production. This hypothesis was reinforced by the finding that addition of FS, to reduce the act A:FS ratio and sequester secreted activin, markedly suppressed (P < 0.001) FSH (3-fold)-, and LR3 IGF-I (2-fold)-induced E2 output.

The variations in oestrogen levels which occur in men with septic shock were determined and analysed in terms of the changes seen in the levels of other steroid hormones of testicular and adrenal origin. The concentrations of the hormones, oestrone (E1), oestradiol (E2), testosterone (T), delta <em>4</em>-<em>androstenedione</em> (delta <em>4</em>), cortisol (F) and progesterone (P<em>4</em>) were determined by radioimmunoassay. The serum levels of cholesterol, triglycerides, phospholipids and non-esterified fatty acids (NEFAs) were also determined. Two groups of male septic shock patients were studied within the first 2<em>4</em> h following the admission to the Intensive Care Unit. Group I (n = 2<em>4</em>) patients died. Group II (n = 22) patients recovered. Both groups were compared to a control group (n = <em>4</em><em>4</em>) of healthy men. In group I patients, serum E1 levels were 3900 +/- 900 pmol/l, 12-fold higher than controls (296 +/- 22 pmol/l) [P less than 0.001], serum E2 levels were 880 +/- 170 pmol/l, 6-fold above control levels (158 +/- 30 pmol/l) [P less than 0.001] and serum T levels were 1.7 +/- 0.3 nmol/l, 11-fold lower than in controls (18.7 +/- 1.9 nmol/l) [P less than 0.001]. Serum P<em>4</em> and F levels were slightly increased (P less than 0.05) and delta <em>4</em> <em>androstenedione</em> levels were unchanged. Groups II serum estrogen levels (81<em>4</em> +/- 350 pmol/l) [P less than 0.01] were higher than controls and serum T levels were 2-3 times less than control levels (5.5 +/- 2 nmol/l) [P less than 0.01]. The group II serum P<em>4</em>, F and delta <em>4</em> <em>androstenedione</em> levels did not differ from control levels. The levels of cholesterol, triglycerides, phospholipids and NEFAs were all decreased to similar, significant, degrees in both groups of shock patients. The dramatic increase in E1 levels associated with the decrease in T suggests an adrenal-testicular relationship with possible potentiation of aromatization of adrenal or testicular androgens in men in septic shock. The determination of serum E1 and T during septic shock in men could form the basis for prognostic estimations of septic shock severity and for a new therapeutic approach to shock.

Some investigators have suggested that "dysregulation" of cytochrome P<em>4</em>50c17 alpha may result in the exaggerated secretion of ovarian androgens in hyperandrogenism. Although the majority of hyperandrogenic (HA) patients demonstrate an ovarian source for their androgens, approximately 50% also display adrenocortical hyperactivity and adrenal androgen excess. To determine whether 17-hydroxylase (17-OH) and/or 17,20-lyase dysregulation is responsible for the adrenocortical abnormalities noted in many HA patients, we studied 92 consecutive women with hirsutism and/or HA oligomenorrhea; 26 healthy eumenorrheic nonhirsute women served as controls. The basal levels of total and free testosterone (T), sex hormone-binding globulin, and dehydroepiandrosterone sulfate were measured, and pregnenolone, 17-hydroxypregnenolone, progesterone, 17-hydroxyprogesterone, dehydroepiandrosterone, and <em>androstenedione</em> were measured 0 and 60 min after the acute iv administration of ACTH-(1-2<em>4</em>). Controls and HA patients did not differ in mean age or body mass, but HA women had higher basal T, free T, dehydroepiandrosterone sulfate, <em>androstenedione</em>, and LH/FSH and lower sex hormone-binding globulin levels. The mean estimated basal 17-OH activity was higher among HA patients than in controls. Although 52 HA patients demonstrated solely an exaggerated basal delta 5-17-OH activity estimate, few HA patients had an exaggerated estimate for either basal delta <em>4</em>-17-OH or ACTH-stimulated 17-OH activity. No HA patient demonstrated an exaggerated 17,20-lyase basal activity, whereas 1<em>4</em> demonstrated an exaggerated delta <em>4</em>-17,20-lyase ACTH-stimulated activity only. There was no association between these estimates of 17-OH and 17,20-lyase activities and the circulating adrenal androgen levels in HA women. Importantly, none of the patients demonstrated an increase in the basal activities of both 17-OH and 17,20-lyase, and only <em>4</em> patients demonstrated an exaggerated ACTH-stimulated activity of both 17-OH and 17,20-lyase. In conclusion, the steroidogenic profile observed in this population of HA women before and after ACTH-(1-2<em>4</em>) stimulation is not consistent with dysregulation of cytochrome P<em>4</em>50c17 alpha and probably represents a generalized alteration of adrenocortical control or biosynthesis.

Congenital lipoid adrenal hyperplasia (lipoid CAH) is the most severe form of CAH, eventually destroying all adrenal and gonadal steroidogenesis. Lipoid CAH is caused by mutations in the steroidogenic acute regulatory protein (StAR), which facilitates the entry of cholesterol into mitochondria to initiate steroidogenesis. Patients with lipoid CAH typically present with a salt-losing crisis in the first 2 months of life, although presentation as late as 10 months with partial retention of StAR activity has been reported. We describe eight patients from six Saudi Arabian families who were first diagnosed at 1-1<em>4</em> months of age (median, <em>4</em>-7 months; mean, 7 months). Five patients were <em>4</em>6,XY, and three were <em>4</em>6,XX. At presentation, all had hyponatremia, hyperkalemia, elevated ACTH, and low cortisol. Pregnenolone, progesterone, 17-hydroxypregnenolone, 17-hydroxyprogesterone, testosterone, <em>androstenedione</em>, and dehydroepiandrosterone sulfate were all low in those patients in whom it was measured. DNA sequencing showed that one patient was homozygous for the StAR mutation M1<em>4</em><em>4</em>R, and the other seven, from five apparently unrelated families, were homozygous for the StAR mutation R182H. Each mutation was recreated in a human StAR cDNA expression vector and found to be wholly inactive in a standard assay of COS-1 cells cotransfected with the cholesterol side-chain cleavage enzyme system. Thus, the loss of all assayable activity in vitro correlated poorly with the later onset of clinical symptoms in these patients. Lipoid CAH may present much later in life than previously thought.

Serum levels of sex-hormones, sex-hormone binding globulin, gonadotropin, and prolactin were evaluated during the follicular and the luteal phases in 65 women with epilepsy and in 20 healthy controls. Twenty-one patients were treated with sodium valproate (VPA), 21 with phenobarbital (PB), and 23 with carbamazepine (CBZ). VPA does not stimulate liver microsome enzymes, whereas PB and CBZ do. Patients on VPA therapy showed higher body weight and body mass index, but no significant differences in hirsutism score, or in ovary volume or polycystic ovary prevalence (at ultrasound examination). Estradiol levels were lower in all patient groups than in healthy controls in the follicular but not in the luteal phases. VPA affected luteal progesterone surge in 63.6% of cases. This effect was significantly lower in the CBZ and PB groups. Furthermore, increases in testosterone and delta <em>4</em>-<em>androstenedione</em> levels and in free androgen index, along with a higher luteinizing hormone-follicle-stimulating hormone ratio in the luteal phase, were observed in women treated with VPA. Although sex-hormone binding globulin levels were higher in CBZ and PB than in VPA-treated patients, the differences were not significant because of the wide dispersion of the carrier protein levels. Inducer antiepileptic drugs decreased dehydroepiandrosterone sulfate levels, which remained unchanged during VPA treatment. No significant differences occurred in basal gonadotropin and prolactin levels.

OBJECTIVE

To document the relative importance of endogenous sex steroids in modulating the frequency of orgasms, the dominant aspect of sexual behaviour in healthy eugonadal men.

METHODS

Measurement of adrenal and testicular sex steroids in a sample of army recruits and study of their relation to frequency of orgasms ascertained by questionnaire after potential confounding variables were controlled for.

METHODS

Military campus and military hospital laboratories in Athens, Greece.

METHODS

92 consecutively enrolled healthy male recruits aged 18-22 years.

METHODS

Weekly number of orgasms. Serum concentrations of testosterone, dehydroepiandrosterone sulphate, dihydrotestosterone, oestradiol, oestrone, delta-<em>4</em>-<em>androstenedione</em>, and sex hormone binding globulin.

RESULTS

Serum dihydrotestosterone concentration was the only independent hormonal predictor of the frequency of orgasms; an increase in concentration of 1.36 nmol/l (about 2 SD) corresponded to an average increase of one orgasm a week.

CONCLUSIONS

Differences in concentrations of circulating dihydrotestosterone within the normal range may represent a major predictor of sexual activity in healthy young men.

A simple and efficient method is described for the purification of microsomal aromatase cytochrome P-<em>4</em>50 from human placenta. The enzyme was solubilized with Emulgen 913 and sodium cholate and subjected to chromatography on a column of Sepharose <em>4</em>B coupled with a specific monoclonal antibody, followed by hydroxyapatite column chromatography. The specific cytochrome P-<em>4</em>50 content of purified aromatase was 13.1 (12-1<em>4</em>.8) nmol/mg of protein. Aromatase assays were carried out with reconstituted systems of bovine liver P-<em>4</em>50 reductase and dilauroyl-L-alpha-phosphatidylcholine with [1 beta-3H,<em>4</em>-1<em>4</em>C]-<em>androstenedione</em> as substrate. The specific activity of purified aromatase was 65.0 (50.6-7<em>4</em>.3) nmol.min-1.(mg of protein)-1 or a turnover rate of 5.0 (<em>4</em>.3-5.9) min-1. The total recovery of purified aromatase activity was 32.2%, and P-<em>4</em>50 recovery was 17.6%. The Km of immunoaffinity-purified aromatase was 12, 210, <em>4</em>1, and 2830 nM for <em>androstenedione</em>, 16 alpha-hydroxy<em>androstenedione</em>, testosterone, and 16 alpha-hydroxytestosterone, respectively. The very high Km value for 16 alpha-hydroxytestosterone aromatization gives a reasonable indication that estriol is not the directly aromatized product in the fetoplacental unit of human pregnancy. The aromatase P-<em>4</em>50 was subjected to SDS-polyacrylamide gel electrophoresis in increasing quantities. Silver stain detection techniques indicated a single band having a molecular mass of 55 kDa with greater than 97% purity. The stability analysis showed a half-life of over <em>4</em> years on storage at -80 degrees C.

Diabetic women may have an increased risk of developing endometrial carcinoma. Ovarian and adrenal activity seem to be factors in the genesis of this cancer. We have measured serum sex hormone-binding globulin (SHBG), free and bound fractions of estrogens and androgens, and gonadotropins in 20 consecutive postmenopausal insulin-treated diabetic women and 16 normal postmenopausal women. The diabetics were nonketoacidotic, without nephropathy and without proliferative retinopathy. The groups were comparable regarding age and percent ideal body weight. The diabetic group had significantly increased serum levels of estrone (P less than 0.001), estrone sulfate (P less than 0.05), 17 beta-estradiol (P less than 0.02), and SHBG (P less than 0.001). Levels of testosterone, delta <em>4</em>-<em>androstenedione</em>, and dehydroepiandrosterone sulfate tended to be higher (not significantly) in the diabetics. FSH and LH levels were similar in the two groups, while serum PRL was significantly lower in the diabetic group (P less than 0.02). The hormonal changes in the diabetics were not related to control of the diabetes. We conclude that total estrogen levels are increased in postmenopausal women with insulin-treated diabetes mellitus. High SHBG levels in these patients tend to keep the free fractions of sex hormones within normal limits.

To determine if delta 1-testololactone can inhibit the peripheral aromatization of <em>androstenedione</em> (delta), nine postmenopausal women with metastatic breast cancer were studied before and after 2 weeks of therapy with 250 mg of the drug, given every 6 h by mouth. The conversion ratio of delta to estrone (E1) was significantly reduced (P less than 0.005) from a mean (+/-SE) of 0.0098 +/- 0.0025 before to 0.0009 +/- 0.0005 after treatment. The drug's effect on the metabolism of delta seemed to be specific since significant changes in the MCR of delta and in the conversion ratio to testosterone were not observed. That this inhibition of peripheral aromatization had an effect on E1 metabolism was shown by the significant decrease (P less than 0.01) of mean serum E1 levels from 22 +/- 3 pg/ml before to 12 +/- 1 pg/ml after treatment. Serum estradiol levels rose slightly from 8 +/- 0.8 to 12 +/- <em>4</em> pg/ml. Serum delta and testosterone levels were unchanged by therapy. These data are consistent with the concept that delta 1-testololactone is a potent inhibitor of peripheral aromatization of delta to E1. This mechanism could explain the antitumor properties of this compound.

The steroidogenic activity of human fetal testes during early and midgestation was monitored by analyzing 58 individual fetal testes (aged 6-20 weeks of pregnancy) for endogenous pregnenolone (P5), progesterone, 17-hydroxyprogesterone, dehydroepiandrosterone, <em>androstenedione</em>, testosterone (T) and estradiol. A clear increase in testicular steroid concentrations, especially in those of T and other 3-keto-<em>4</em>-ene steroids, occurred between 8-11 weeks of gestation and reached maximum between 11-1<em>4</em> weeks. The three steroids present in highest concentrations were P5, <em>androstenedione</em>, and T (maximum concentrations, 1.9-2.7 ng/mg wet tissue). The levels of all of the C-19 steroids measured decreased clearly between weeks 1<em>4</em>-20 of gestation, whereas those of the C-21 steroids, P5, progesterone, and 17-hydroxyprogesterone, remained relatively high. Our results suggest that the metabolic reactions converting P5 to androgens are activated in the human fetal testis within a short time range between 8-11 weeks of gestation. The increased androgen production is possibly a consequence of increased 3 beta-hydroxysteroid dehydrogenase activity. Testicular androgen production decreases in the beginning of the second trimester of pregnancy, most likely due to a blockade in the C-21 steroid side-chain cleavage.

Metabolites play essential roles in biological systems, but detailed identities and significance of the seminal plasma metabolome related to bull fertility are still unknown. The objectives of this study were to determine the comprehensive metabolome of seminal plasma from Holstein bulls and to ascertain the potential of metabolites as biomarkers of bull fertility. The seminal plasma metabolome from 16 Holstein bulls with two fertility rates were determined by gas chromatography-mass spectrometry (GC-MS). Multivariate and univariate analyses of the data were performed, and the pathways associated with the seminal plasma metabolome were identified using bioinformatics approaches. Sixty-three metabolites were identified in the seminal plasma of all bulls. Fructose was the most abundant metabolite in the seminal fluid, followed for citric acid, lactic acid, urea and phosphoric acid. <em>Androstenedione</em>, <em>4</em>-ketoglucose, D-xylofuranose, 2-oxoglutaric acid and erythronic acid represented the least predominant metabolites. Partial-Least Squares Discriminant Analysis (PLSDA) revealed a distinct separation between high and low fertility bulls. The metabolites with the greatest Variable Importance in Projection score (VIP>> 2) were 2-oxoglutaric acid and fructose. Heat-map analysis, based on VIP score, and univariate analysis indicated that 2-oxoglutaric acid was less (P = 0.02); whereas fructose was greater (P = 0.02) in high fertility than in low fertility bulls. The current study is the first to describe the metabolome of bull seminal plasma using GC-MS and presented metabolites such as 2-oxoglutaric acid and fructose as potential biomarkers of bull fertility.

OBJECTIVE

Although androgenic alopecia is recognised to be a symptom of polycystic ovary syndrome (PCOS), it is not known whether polycystic ovaries (PCO) and associated endocrine abnormalities are present in patients who present with alopecia as a primary complaint. We therefore set out to determine the strength of the association between androgenic alopecia and PCO. We examined the prevalence of ultrasound-based polycystic ovarian morphology and associated clinical and biochemical features in a large multiethnic group of women whose presenting complaint was of alopecia, and in a control group.

METHODS

We studied 89 women of mixed ethnic origin with androgenic alopecia and compared them to 73 control women. A detailed history was taken, anthropometry was performed and assessment of body-hair distribution was made. The presence of PCO was established by pelvic ultrasound scan. Serum gonadotrophins, testosterone, androstenedione, dihydrotestosterone and sex hormone binding globulin concentrations were measured.

RESULTS

Women with alopecia had a higher prevalence of PCO and hirsutism than the control population (PCO: 67% vs 27%, P<0.00001; hirsutism: 21% vs 4%, P=0.003). Women with alopecia (with or without PCO) had higher testosterone, androstenedione and free androgen index than controls, even though few had frankly abnormal androgens.

CONCLUSIONS

These findings confirm an association between androgenic alopecia and PCO, and other symptoms of hyperandrogenaemia. Thus most women who present with androgenic alopecia as their primary complaint also have PCO and have indices of abnormal androgen production. Since PCO is a well known risk factor for development of type 2 diabetes, this association has important implications for long-term management.

OBJECTIVE

To examine whether follicle loss due to ovarian aging is responsible for the occurrence of regular menstrual cycles in aging women with polycystic ovary syndrome (PCOS), the size of the FSH-sensitive follicle cohort was estimated by the exogenous follicle-stimulating hormone ovarian reserve test (EFORT) and related to the follicle count as measured by ultrasound.

METHODS

Prospective study.

METHODS

Reproductive endocrinology unit of an academic medical center.

METHODS

Twenty-seven aging women with PCOS (35.8-<em>4</em>9.<em>4</em> years): 20 with regular menstrual cycles and 7 with oligomenorrhea or amenorrhea.

METHODS

EFORT and transvaginal ultrasound.

METHODS

Baseline (cycle day 2, 3, or <em>4</em>) FSH, <em>androstenedione</em> (A), T, E(2), and inhibin B levels, the E(2) and inhibin B increment after the EFORT, and the follicle count.

RESULTS

After correction for the body mass index (BMI), the inhibin B increment was higher in the irregular menstrual group, but the E(2) increment did not differ significantly between the two groups. Ultrasound showed a median follicle count of 8.5 (<em>4</em>.0-18.0) in women with regular menstrual cycles (n = 16), compared with 18.0 (8.0-35.0) in irregularly menstruating women (n = 7). The follicle count was significantly correlated to the FSH-induced E(2) increment (r = 0.656) as well as to the inhibin B increment (r = 0.65<em>4</em>). The regularly menstruating group was significantly older, had a higher basal FSH concentration, and had lower androgens than the irregularly menstruating group.

CONCLUSIONS

The smaller follicle count, the older age, the higher FSH concentration, and the lower FSH-induced inhibin B increment found in women with PCOS and a regular menstrual cycle confirm that a decrease in the size of the follicle cohort due to ovarian aging is largely responsible for the regular menstrual cycles in aging PCOS women.

BACKGROUND

Recent studies challenge the assumption that hypogonadism in Prader-Willi syndrome (PWS) is due only to hypothalamic dysfunction.

OBJECTIVE

The aims of the study were to characterize sexual development and reproductive hormones in PWS males and investigate the etiology of hypogonadism.

METHODS

Physical examination and blood sampling were performed on 37 PWS males, ages <em>4</em> months to 32 yr.

RESULTS

All had a history of undescended testes; age at orchiopexy ranged from 2 months to 6 yr. Pubertal signs were variable, but none achieved full genital development. Anti-Mullerian hormone (AMH) levels in PWS boys were near the lower limits of normal, decreasing from <em>4</em><em>4</em>.<em>4</em> +/- 17.8 ng/ml (mean +/- sd) in young children to 5.9 +/- <em>4</em>.7 ng/ml in adolescents, similar to normal males. In contrast, inhibin B was consistently low (27.1 +/- 36.1 pg/ml) or undetectable in all age groups. In adult males, FSH levels were high (20.3 +/- 18.3 IU/liter), LH levels were normal (<em>4</em>.2 +/- <em>4</em>.3 IU/liter), and testosterone levels were low (1.87 +/- 1.17 ng/ml). Only two adults had severe hypogonadotropic hypogonadism with undetectable levels of LH and FSH and high AMH levels (3<em>4</em>.9 and 36.7 ng/ml), unlike the other nine adults with AMH levels 2.6 +/- 2.1 ng/ml. Androstenedione (1.06 +/- 0.30 ng/ml) and DHEAS (281.1 +/- 1<em>4</em>3.6 microg/dl) in adult PWS were normal.

CONCLUSIONS

Pubertal development in PWS is characterized by normal adrenarche, variable hypothalamic dysfunction, and hypogonadism due to a unique testicular defect. Primary testicular dysfunction is a major component of hypogonadism in PWS.

OBJECTIVE

To evaluate female sexual dysfunction (FSD) in obese women in comparison with age-matched control group, emphasising their hormonal and psychological status.

METHODS

Sixty-four sexually active obese premenauposal women were compared with a control group of 27 age-matched healthy volunteers with a normal body mass index (BMI). The obese group was evaluated in three categories according to BMI (kg/m(2)), as group 1 (30-3<em>4</em>.9), group 2 (35-39.9) and group 3 >> <em>4</em>0). All women were evaluated with Female Sexual Function Index (FSFI) questionnaire and Beck Depression Inventory (BDI). In addition, serum levels of follicle-stimulating hormone, luteinizing hormone, prolactin, dehydroepiandrosterone-SO(<em>4</em>) , free testosterone, 17α-hydroxyprogestrone, <em>androstenedione</em>, oestradiol, free thyroxine and thyrotropin were determined.

RESULTS

The mean FSFI scores were not statistically significant between control and obese patients (P = 0.29). FSD was diagnosed in 50% (32/6<em>4</em>) and <em>4</em>1% (11/27) of the patients in the obese and control groups, respectively (P = 0.3<em>4</em>). There were no differences between total FSFI and FSFI domain scores among BMI categories. BDI scores were significantly higher in the obese groups than in healthy controls, and negatively correlated with total FSFI and all FSFI domain scores. Among hormonal analyses, only free testosterone levels were negatively correlated with total FSFI scores.

CONCLUSIONS

This study showed that obesity has no significant relationship with FSD, but obese patients were found to be in a more depressive mood than age-matched normal counterparts.