Rainbow trout fry were treated with equimolar quantities of cortisol, cortisone, <em>androstenedione</em>, and 11 beta-hydroxy<em>androstenedione</em>, added to the aquarium water during a <em>4</em>-week period. All four steroids inhibited ovarian growth. <em>Androstenedione</em> did not influence gonadal sex differentiation; the other steroids pushed the sex ratio in the male direction. However, a near to normal sex ratio was observed 300 days after the 11 beta-hydroxy<em>androstenedione</em> treatment. When added to the food in two different doses during 8 weeks, 11 beta-hydroxy<em>androstenedione</em> had a pronounced masculinizing effect, and <em>androstenedione</em> did not modify the gonads at all. It is suggested that the two exogenous corticosteroids were probably converted into 11-oxygenated <em>androstenedione</em> derivatives, and that these 11-oxygenated <em>androstenedione</em> derivatives are particularly important in sustaining the differentiation and early development of the testes in rainbow trout. Testosterone does not seem to be indispensable for these processes, because two different doses of testosterone-blocking cyproterone acetate, added to the food for 8 weeks, failed to affect early gonad development.

Eight patients with congenital microphallus were investigated. Plasma luteinizing hormone, follicle-stimulating hormone, testosterone and <em>androstenedione</em> levels were obtained in all cases. In addition, the response to the administration of human chorionic gonadotropin, luteinizing horomone-releasing hormone and adrenocorticotropic hormone, the assessment of testicular histology by electron microscopy and the measurement of dihydrotestosterone formation by preputial skin were determined in some patients. The results of these studies were compared to similar studies in 6 normal prepubertal boys, <em>4</em> boys with bilateral cryptorchidism, 1 male infant with anorchia and 1 adult with hypogonadotropic hypogonadism. The clinical and endocrinological findings in the 8 patients with microphallus can be divided into 2 distinct categories. In 5 patients the disorder is familial, gonadotropin levels are low and there is a normal response to stimulation with chorionic gonadotropin. The data are compatible with the possibility that 3 (possibly 5) of the 8 patients with microphallus have hypogonadotropic hypogonadism. In the other group the cases are sporadic, serum luteinizing hormone and follicle-stimulating hormone levels are elevated and plasma testosterone failed to increase after short-term treatment with chorionic gonadotropin. In these patients a primary testicular disorder appears to be responsible. Experimental and clinical evidence suggests that microphallus results from defective testicular function during the second and third trimesters of pregnancy, either as the result of defective gonadotropin secretion or defective androgen synthesis.

OBJECTIVE

A previously published study has identified that anovulatory women with PCOS have an increased response of 17 alpha-hydroxyprogesterone (17OHP) and androstenedione to a GnRH analogue suggesting dysregulation of cytochrome P450c17 alpha. The object of this study was to compare the responses of the pituitary-ovarian axis to a single dose of a long-acting GnRH agonist (GnRHa) in both ovulatory and anovulatory women with PCOS with those in normal subjects.

METHODS

Comparative study of responses of LH, FSH and ovarian steroids to buserelin and the adrenal steroid response to synthetic ACTH in two groups of women with hyperandrogenaemia and polycystic ovaries: those with anovulatory menses or amenorrhoea and those with equally elevated serum testosterone concentrations but regular menses. Results in both groups of women with PCO were compared with those in normal subjects.

METHODS

Twenty-four women with hyperandrogenism and PCO (14 had oligo or amenorrhoea, 10 regular cycles) and 12 weight matched controls with normal ovaries, regular cycles and neither clinical nor biochemical evidence of hyperandrogenism. Subjects were given synthetic ACTH (Synacthen) 250 micrograms i.v. on day 1 of the study and blood collected at 30 and 60 minutes thereafter. On the evening of day 1, dexamethasone treatment was commenced to suppress adrenal androgens. GnRHa 100 micrograms s.c. was given on day 2 and blood samples collected at 30-minute intervals for 4 hours and once more at 24 hours after the injection.

RESULTS

The acute responses of both immunoactive and bioactive LH to GnRHa were significantly greater in the ovulatory PCO group (ovPCO) than controls but the response was greater in anovulatory women with polycystic ovaries (anovPCO) than in either ovPCO or controls, throughout the 24-hour study period. Despite the discrepancy in LH concentrations, basal serum concentrations of androstenedione were equally elevated in anovulatory and ovulatory women with PCO, compared with controls. There was a small but significant increase in androstenedione following GnRHa in both PCO groups at 24 hours but not in controls. A similar pattern was observed in the response of 17OHP to GnRHa although the response was significantly higher than controls in anovPCO women only. By contrast, the responses of both androstenedione and 17OHP to 250 micrograms synthetic ACTH were similar in PCO women to those in controls.

CONCLUSIONS

These data provide evidence for ovarian hypersecretion of androgens in ovulatory, as well as anovulatory women with PCO, supporting the concept of abnormal regulation of 17-hydroxylase and (17,20-lyase activity in the ovary. The finding of an equal degree of hyperandrogenaemia in ovPCO and anovPCO groups, even though LH levels were much higher in the latter, suggests that hypersecretion of LH is not the primary cause of ovarian hyperandrogenism. Hyperandrogenism in PCOs may therefore represent an intrinsic abnormality of ovarian theca-interstitial cell function.

To study ovarian and adrenal steroid profiles of women with idiopathic hirsutism, we compared sex steroid and basal and corticotropin (ACTH)-stimulated adrenal steroid levels before and after ovarian suppression induced by a long-acting gonadotropin-releasing hormone agonist analog (GnRH-a) in 2<em>4</em> hirsute women without hyperandrogenemia. Twelve healthy women served as controls for basal and ACTH-stimulated adrenal steroid levels. Serum levels of testosterone (T), sex hormone-binding globulin (SHBG), estradiol (E2), basal and ACTH-stimulated 17-hydroxyprogesterone (17OHP), dehydroepiandrosterone (DHEA), DHEA sulfate (DHEAS), delta <em>4</em>-<em>androstenedione</em> (delta <em>4</em>-A), 11-deoxycortisol (S) and cortisol (F), and basal and luteinizing hormone-releasing hormone (LHRH)-stimulated gonadotropin levels were measured before and 21 days after 3.75 mg intramuscular triptorelin in hirsute women. Basal T levels and basal and ACTH-stimulated delta <em>4</em>-A, DHEA, and DHEAS levels were not different in hirsute women with respect to controls. Basal and ACTH-stimulated 17OHP was elevated, and decreased to normal after ovarian suppression with triptorelin. Although basal and ACTH-stimulated delta <em>4</em>-A levels were normal, the delta delta <em>4</em>-A/delta F and delta delta <em>4</em>-A/delta 17OHP ratios were elevated and remained elevated after ovarian suppression, suggesting enhanced adrenal delta <em>4</em>-17,20-lyase activity. T, F, S, and DHEAS levels were not affected by ovarian suppression. Basal and ACTH-stimulated 17OHP and delta <em>4</em>-A, and stimulated DHEA concentrations were reduced with ovarian suppression, but their net increment and ratio to the increase of F in response to ACTH remained unchanged, reflecting the ovarian contribution to the secretion of these steroids. We conclude that idiopathic hirsute women with normoandrogenemia show an increase in ovarian secretion of 17OHP and a minimally increased adrenal delta <em>4</em>-17, 20-lyase activity, suggesting that mild forms of ovarian and adrenal functional hyperandrogenism may be present in these patients with otherwise unexplained hirsutism.

The serum levels of the following steroids were measured in 59 hirsute patients before (control) and after (post-dexamethasone) administration of dexamethasone (Dex) for 7 days: cortisol (F), dehydroepiandrosterone (DHEA), its sulfate (DHEA-S), <em>androstenedione</em> (A), testosterone (T), and 5alpha-dihydrotestosterone (DHT). Assuming that Dex-suppressibility implied adrenal origin, the source of excess androgens was also evaluated. All patients showed elevated level of only one androgen: <em>4</em> had elevated DHEA-S; <em>4</em> had elevated T; 3 had elevated DHT. No patient had an elevated DHEA or A without an evelation of the other androgens. The control levels of DHEA-S were above normal in <em>4</em>5 patients; the DHT levels were elevated in <em>4</em>3 patients; 31 patients had elevated T levels; and 25 patients elevated A; and 2<em>4</em> patients, DHEA levels. In 32 patients with adequate suppression of adrenal androgens after 7 days of Dex administration, the source of excess androgens could be evaluated. Of 13 patients with elevated A levls, the excess A production was of adrenal origin in 6 cases, of ovarian origin in 5 cases, and of mixed origin in 2 cases. Of 15 patients with elevated T levels, the excess T production was of adrenal origin in 3 cases, of ovarian origin in 10 cases, and of mixed origin in the remaining 2 cases. Of 25 patients with elevated DHT levels, the excess DHT production was of adrenal origin in 16 patients, of ovarian origin in 5 patients, and of mixed origin in <em>4</em> patients. Of the 32 patients with an adequate Dex suppression test, 1<em>4</em> showed evidence of adrenal hyperandrogenism, 5 had ovarian hyperandrogenism, and mixed hyperandrogenism was present in the remaining 13 patients. There was an adrenal source of hyperandrogenism in 27 of 32 patients (1<em>4</em> pure adrenal and 13 mixed adrenal-ovarian), which represents 85% of the 32 patients.

OBJECTIVE

To analyse the clinical characteristics and relevant hormonal profile in type 1 diabetic patients with and without ED.

METHODS

Fifty one type 1 diabetic patients were studied. ED was assessed by direct interview. Chronic diabetic complications, smoking and alcohol status as well as current use of medications were recorded. Hormonal profile consisted of plasma LH, FSH, prolactin, <em>androstenedione</em> (Delta(<em>4</em>)), dehydroepiandrosterone (DHEA), DHEA-sulfate (DHEA-S), free testosterone (FT), estradiol (E(2)), sex hormone binding globulin (SHBG), dihydrotestosterone (DHT), cortisol, TSH and free thyroxine (FT(<em>4</em>)).

RESULTS

ED was present in 2<em>4</em> patients (<em>4</em>7%) (group 1), who were older (P<0.001), had a longer diabetes duration (P<0.001) and a higher systolic blood pressure (P=0.017) when compared to the subjects who did not complain (group 2). ED was positively correlated to all diabetes-related complications (P<0.02). Antidepressive drug(s) were more frequent in group 1 (P=0.007), as well as prokinetics (P=0.0<em>4</em>3) and ACE-inhibitors (P=0.010). HbA(1)c was comparable. Patients with ED had lower levels of Delta(<em>4</em>) (P=0.003), DHEA (P<0.001), DHEA-S (P=0.002), FT (P=0.08) while SHBG (P=0.010) and LH (P=0.022) were higher compared to group 2. Multiple logistic regression analysis showed an independent association of ED with Delta(<em>4</em>) (P=0.016), DHEA-S (P=0.037), SHBG (P=0.001) and insulin dose (P=0.025). There was no significant difference for all other measured hormones.

CONCLUSIONS

ED is impressively prevalent in type 1 diabetes and is associated with age, diabetes duration, chronic complications and decreased androgens.

Standardized bilateral ovarian-adrenal vein catheterization was utilized to assess directly glandular steroid release in 60 androgenized women without evidence of a functional neoplasm. Testosterone (T), dihydrotestosterone (DHT), <em>androstenedione</em> (delta <em>4</em> A), dehydroepiandrosterone (DHEA), DHEA sulfate (DHEA-S), 17-hydroxyprogesterone (17-OHP), and cortisol (F) were measured by radioimmunoassay in samples obtained from a peripheral vein and the four glandular veins (all values are given as nanograms per milliliter, mean +/- standard deviation). Peripheral values were as follows: T, 0.68 +/- 0.<em>4</em>3; DHT, 0.32 +/- 0.13; delta <em>4</em> A, 2.2 +/- 2.0; DHEA, 8.8 +/- 8.9; DHEA-S, 3137 +/- 177<em>4</em>; 17-OHP, 2.0 +/- 3.0; and F, 216 +/- 121. Peripheral elevations of at least one androgen were found in 80% of the 60 cases (T, 38%; DHT, 18%; delta <em>4</em> A, 50%; DHEA, <em>4</em>5%; and DHEA-S, 37%). Ovarian-peripheral vein gradients ( OPGs ) and adrenal-peripheral vein gradients ( APGs ) served as semiquantitative estimates of glandular secretion. OPGs were as follows: T, 0.<em>4</em> +/- 1.1; DHT, 0.1 +/- 0.2; delta <em>4</em> A, 3.<em>4</em> +/- 7.0; DHEA, 1<em>4</em>.6 +/- 100; DHEA-S, -288 +/- 523; 17-OHP, <em>4</em>.5 +/- 8.<em>4</em>; and F, -35 +/- <em>4</em>7. APGs were as follows: T, 0.88 +/- 1.3; DHT, 1.1 +/- 0.9; delta <em>4</em> A, 1<em>4</em>.<em>4</em> +/- 38.<em>4</em>; DHEA, 327 +/- 367; DHEA-S, 85<em>4</em> +/- 1223; 17-OHP, 20.8 +/- <em>4</em>1.3; and F, 1252 +/- 2023. Excess ovarian and/or adrenal androgen output was assumed in a given individual when one or more of the respective T, DHT, delta <em>4</em> A, DHEA, and DHEA-S gradients exceeded the upper 95% confidence limits of normal previously established in this laboratory.(ABSTRACT TRUNCATED AT 250 WORDS)

Stimulation of rat granulosa cell aromatase activity by FSH has recently been used as a sensitive biological end point to develop an in vitro FSH bioassay. The present report provides a detailed validation and application of this assay. In the presence of <em>androstenedione</em> and diethylstilbestrol, FSH stimulated estrogen production in a dose-dependent manner. Although addition of high doses of a phosphodiesterase inhibitor [1-methyl-3-isobutyl xanthine (MIX)] decreased maximal estrogen production, treatment with 0.125 mM MIX increased the sensitivity of granulosa cells to FSH, presumably by minimizing endogenous cAMP breakdown. Addition of insulin and human CG (hCG) further synergistically enhanced granulosa cell sensitivity to FSH. Although inclusion of gonadotropin-free serum obtained from hypophysectomized male rats decreased the assay sensitivity, pretreatment of serum with polyethylene glycol [(PEG) 10-1<em>4</em>%] resulted in a dose-dependent decrease in the serum-interfering effect. Studies using exogenous [125I]iodo-rat FSH or RIA measurement indicated recovery of 9<em>4</em>-98% FSH after pretreatment of serum with 12% PEG. In the presence of the PEG-pretreated gonadotropin-free serum (<em>4</em>%), ovine, rat, and human FSH preparations induced parallel dose-response curves for estrogen production with minimal detectable doses of 0.12 ng, 0.12 ng, and 0.12 mIU/culture, respectively. In contrast, treatment with GH, PRL, TSH, and ACTH did not affect estrogen production. The apparent stimulatory effect of high doses (greater than 60 ng/culture) of LH and hCG could be attributed to FSH contamination or intrinsic FSH activity in these preparations. Changes in serum bioactive FSH levels were studied in adult male rats after GnRH administration. GnRH (5 micrograms/rat) treatment significantly elevated FSH levels within 30 min after injection. Maximal increases (approximately 2.8-fold) in serum bioactive FSH were observed between 60-120 min. At 8 h after treatment, FSH levels decreased to control levels. Comparison between granulosa cell aromatase bioassay and RIA results indicated no apparent changes in the bio- to immuno- ratio of FSH after GnRH treatment. In conclusion, extreme sensitivity of the bioassay allows the measurement of circulating levels of bioactive FSH. Since rat granulosa cells respond to FSH preparations from different species, the in vitro assay should also provide valuable information on FSH levels in many animal species including those lacking a specific RIA. Measurement of serum levels of bioactive FSH should provide insight regarding the role of FSH in various physiological and pathological conditions.

Androgen metabolism (<em>androstenedione</em> and testosterone) has been assessed in the digestive tube and gonads of the echinoderm Paracentrotus lividus exposed to different concentrations of the biocide triphenyltin (TPT) in a semi-static water regime for <em>4</em> weeks. Key enzymatic activities involved in both synthesis and metabolism of androgens, namely 17beta-hydroxysteroid dehydrogenases (17beta-HSDs), 3beta-HSDs, 5alpha-reductases, P<em>4</em>50-aromatase, palmitoyl-CoA:testosterone acyltransferases (ATAT) and testosterone sulfotransferases (SULT), were investigated in digestive tube and/or gonads of control and TPT-exposed specimens in an attempt to see whether androgen metabolism was altered by exposure. In agreement with previous data for vertebrates, exposure to TPT led to a concentration dependent decrease of P<em>4</em>50-aromatase that was statistically significant at the highest TPT concentration tested (225ng/L). Additionally, increased metabolism of testosterone to form dihydrotestosterone (DHT) and 5alpha-androstane-3beta,17beta-diol was observed, suggesting increased 5alpha-reductase activity in the gonads of TPT-exposed individuals. Interestingly, exposure to TPT induced testosterone conjugating activities in organisms exposed to medium (SULT) and high (ATAT and SULT) TPT concentrations. Despite the changes of androgen metabolizing enzymes, testosterone levels in gonads remained rather stable. In contrast, an increase in testosterone and a concomitant decrease in estradiol were observed in the coelomic fluid of TPT-exposed organisms. Overall, the data indicate the ability of TPT to modulate androgen metabolism and circulating steroid levels in P. lividus and suggest the existence of regulatory mechanisms to maintain stable endogenous levels of testosterone in gonads. This study also contributes to a better knowledge of echinoderm endocrinology.

In this cross-sectional study, plasma levels of dehydroepiandrosterone sulfate (DHEA-S), dehydroepiandrosterone (DHEA), delta <em>4</em>-<em>androstenedione</em> (delta <em>4</em>) and testosterone (T) were measured by RIA in 232 normal subjects of both sexes, aged 2 weeks to 20 years. The results were analyzed in relation to chronological age, body surface and pubertal stage. High levels of plasma androgens were found in newborn infants of both sexes. After 3 months of age, androgen levels were uniformly low and rose with increasing chronological age and body surface. The first significant increase in mean androgen levels was found for DHEA-S. It occurred after 6 years of age in girls and after 8 years in boys. DHEA and T rose in both sexes after 8 years of age. delta <em>4</em> increased steadily with chronological age and body surface in both sexes. When androgen levels were related to body surface, a first significant increase was observed above 1.00 m2 for the four androgens, in both boys and girls. Above 1.20 m2 and 12 years of age, girls had higher mean levels of DHEA-S, DHEA and delta <em>4</em>, but lower mean T levels than boys of the same body surface and chronological age. Before puberty, a positive correlation was found in both sexes between the plasma androgen levels on the one hand, and both chronological age and body surface on the other. Plasma androgen levels markedly increased at stage P2 in both sexes, and further increased with pubertal development. During puberty, girls had higher plasma delta <em>4</em>, but lower plasma T levels than boys of the same pubertal stage. Plasma DHEA-S and DHEA levels were similar in both sexes. In contrast to the plasma androgens, plasma cortisol levels did not show any change in relation either to chronological age or to body surface or pubertal development. Body surface appears to be as good a discriminating factor as chronological age, at least in young children. It also appears from this study that DHEA-S is a good guide for the clinical evaluation of adrenal maturation and may be very useful in evaluating patients with growth or pubertal disturbances.

Adrenal androgen production and adrenocortical biosynthesis are frequently abnormal in women with hyperandrogenism. It is possible that these abnormalities result from hypertestosteronemia of ovarian origin. We prospectively studied the effect of prolonged testosterone (T) administration on adrenocortical biosynthesis and basal hormone levels in seven oophorectomized euandrogenic volunteers, who received 20 mg T in oil (USP), im, three times weekly for 3 weeks. Blood was sampled before T administration (week 0), during therapy (weeks 1, 2, and 3), and 1 week after discontinuation of the medication (week <em>4</em>). Acute adrenal stimulation using 250 micrograms ACTH-(1-2<em>4</em>), iv, was performed on weeks 0, 2, and 3. Circulating T rose from an average of 1.53 nmol/L to 27.9 nmol/L by the end of week 3. Both LH and FSH decreased with T administration, while PRL rose slightly during the initial weeks of T therapy. Little change was observed in the basal levels of 17-hydroxyprogesterone, 11-deoxycortisol, 17-hydroxypregnenolone, estrone, and estradiol. <em>Androstenedione</em> (A) rose concurrent with T administration and normalized after therapy discontinuation. Dehydroepiandrosterone (DHA) decreased significantly with T administration, while DHA sulfate (DHS) increased, leading to an increased DHS/DHA ratio. Total cortisol (F) decreased during T therapy; however, corticosteroid-binding globulin levels also decreased, and the unbound F level remained unchanged. No significant change in adrenocortical response (measured by either the absolute poststimulation level or the net increment in steroid levels) for any steroid was noted after T administration. Although the absolute poststimulation level of A was higher during T administration, this change reflected the increased basal A levels, since the increment in A from 0-60 min was not different. In conclusion, raising endogenous T levels in oophorectomized healthy women 10- to 15-fold for a period of 3 weeks produced a decrease in circulating total F and corticosteroid-binding globulin levels, while maintaining normal levels of unbound F. Although the very high T levels produced in this study resulted in an increased DHS/DHA serum ratio, it remains to be proven that the lesser degree of hypertestosteronemia usually observed in hyperandrogenic patients is sufficient to alter DHS metabolism. With this exception, the adrenocortical biosynthetic abnormalities frequently noted in hyperandrogenism do not appear to result from the elevated circulating T levels.

The concentration of the adrenal steroids dehydroepiandrosterone (DHA) and dehydroepiandrosterone sulfate (DHA-S) was measured by radioimmunoassay in plasma of 13<em>4</em> baboons (Papio cynocephalus) from 0 to 17 yr of age. Plasma <em>androstenedione</em> (delta <em>4</em>-A) was also measured in 289 baboons encompassing the same age range. The concentration of DHA was persistently high from birth to adulthood, exceeding 700 ng/dl at all ages. This differs markedly from the human pattern in which DHA concentrations fall below 50 ng/dl following regression of the fetal adrenal. Plasma DHA-S declined rapidly during the first 2 yr of life, causing a significant age-related decline in the ratio of DHA-S:DHA, a pattern not seen in man. Plasma delta <em>4</em>-A declined during the first 3 yr, but remained greater than 100 ng/dl. delta <em>4</em>-A then rose sharply at age <em>4</em>, the time of gonadal maturation in the baboon. Thus, the baboon differs from man primarily in the persistent elevation of plasma DHA and delta <em>4</em>-A prior to puberty. It remains to be determined whether this difference reflects fundamentally different mechanisms regulating adrenal androgen secretion or merely the closer temporal juxtaposition of fetal adrenal regression and adrenarche due to the more rapid sexual maturation of the baboon.

OBJECTIVE

Flutamide is a non-steroid antiandrogen that specifically blocks the androgen receptor. We have investigated the effect of flutamide treatment on the adrenal androgen response to corticotrophin releasing factor (CRF) in eight patients with polycystic ovary syndrome (PCOS).

METHODS

Eight women with moderate to severe hirsutism, ranging in age from 19 to 23 years were enrolled in the study. Basal hormonal pattern showed anovulatory cycles, increased concentrations of LH, androstenedione and testosterone and increased LH/ FSH ratio. A baseline ultrasound scan revealed polycystic ovaries in all patients. Each received 250 mg of Flutamide twice a day for 6 months.

METHODS

Before treatment and at the end of the sixth month, women were evaluated for hirsutism score and a CRF test was performed to evaluate ACTH, cortisol and adrenal androgen responses.

RESULTS

Androstenedione (delta 4), DHEA-S, 17-hydroxy-progesterone, testosterone and free-testosterone showed significantly reduced responses after six months of flutamide therapy whereas ACTH and cortisol response were similar to those before treatment. Clinical improvement in the degree of hirsutism was observed in all patients. The Ferriman-Gallwey scores decreased from a mean of 22 +/- 2 to 8 +/- 1.5.

CONCLUSIONS

Flutamide induces a significant reduction in adrenal androgen response to the CRF test but not in the response of ACTH and cortisol. The finding that flutamide does not alter the pituitary-adrenal axis shows that flutamide acts by reducing adrenal androgens. These results demonstrate that flutamide is not only effective in the treatment of hirsutism but also reduces adrenal androgen secretion.

To summarize the endocrine approach for the treatment of BPH: much clinical data have accumulated over the past forty years. Until recently, scientists and physicians mainly concentrated on the reduction of androgens as a possible solution. We have come a long way from surgical castration, through the administration of hormones such as estrogen and progesterone, gonadotropin-releasing hormone agonists to the inhibition of an enzymatic reaction reducing testosterone to DHT--the now recognized active intracellular androgen metabolite. Recently, the role of estrogens has been emphasized with the finding that stromal hyperplasia is the main change occurring in BPH. Lately, research has been initiated to examine the clinical effect aromatase inhibitors would have in the treatment of human BPH. Since there is enough evidence that both the epithelial and stromal components of the human prostate undergo hyperplasia in BPH, and individuals vary with respect to their relative epithelial/stromal components, both structures would have to be reduced for therapy to be successful. Therefore, the combination of an antiandrogenic and antiestrogenic effect is theoretically promising. Indeed, prostates of beagles shrunken after treatment with an aromatase inhibitor, further decreased in weight after additional treatment with cyproterone acetate, an antiandrogen. We are now approaching the stage where these "antihormones" are both enzyme inhibitors with actually no side effects that preclude the use of the earlier generation's "antihormonal" hormonal drugs. Furthermore, it has recently been reported that the aromatase inhibitor, <em>4</em>-hydroxy-<em>androstenedione</em> also inhibits human prostatic 5-alpha reductase, at least in vitro. The in vivo relevance of this finding awaits further classification. Thus, a good hormonal treatment that will be both scientifically sound, and clinically safe and effective, seems feasible in the near future. Two main factors have encouraged our interest and research into methods of inhibiting prostatic growth or reducing its obstructive symptomatology: the enormous cost of prostatic operations for outlet obstruction secondary to BPH, and the natural aging process of the population accompanied by deteriorated health precluding anesthesia and prostatic surgery. Medical treatment of BPH has to result in symptomatic improvement, elimination of residual urine, and improvement of flow to be considered successful. These are usually accomplished by surgery and results at least as good as those obtained by operation should be aimed at, if medical treatment is to replace surgery. Although indications for surgery and outcome of operations are well-defined, this is not the case when alternatives to prostatectomy are chosen.(ABSTRACT TRUNCATED AT <em>4</em>00 WORDS)

OBJECTIVE

A combination of flutamide (Eulexin) or nilutamide (Anandron) with a luteinizing hormone-releasing hormone (LHRH) agonist or orchiectomy is the only therapy demonstrated to prolong life in prostate cancer. Recently, the low 50-mg daily dose of Casodex, an analogue of the pure antiandrogen flutamide, was chosen for clinical studies on the basis that the compound was 5 to 10 times more potent than flutamide, as suggested by data obtained in the inappropriate intact rat model. The present study was designed to compare the in vitro antiandrogenic activity of OH-flutamide (OH-FLU), the active metabolite of flutamide, Casodex, and nilutamide.

METHODS

The effect of the antiandrogens was tested on two androgen-sensitive parameters, namely proliferation of the SEM-107 clone of Shionogi mouse mammary tumor cells and secretion of the GCDFP-15 (gross cystic disease fluid protein 15 kDa) in T-<em>4</em>7D and ZR-75-1 human breast cancer cells.

RESULTS

The twofold stimulation of Shionogi cell proliferation caused by a 10-day exposure to 1 nM testosterone was competitively reversed by incubation with OH-FLU, Casodex, or nilutamide, at the respective IC50 values of 72, 2<em>4</em>3, and <em>4</em>12 nM. Moreover, the marked increase in GCDFP-15 release induced by 1 nM testosterone was blocked by OH-FLU. Casodex, or nilutamide at respective IC50 values of 29, 180, and 87 nM in T-<em>4</em>7D cells and at 35, 1<em>4</em>2, and 75 nM in ZR-75-1 cells. Similar data were detected in <em>4</em>-<em>androstenedione</em>-induced Shionogi cell proliferation and in dihydrotestosterone-induced GCDFP-15 secretion in T-<em>4</em>7D cells.

CONCLUSIONS

OH-FLU is 3.1- to 7.8-fold more potent than Casodex, as measured on two in vitro androgen-sensitive parameters, in agreement with our recent in vivo data obtained in the model of castrated rats supplemented with <em>4</em>-<em>androstenedione</em> implants, in which threefold greater potency of flutamide was observed. The present data, as well as other data from the literature, strongly indicate the need to choose a more appropriate dose of Casodex for the treatment of prostate cancer.

Sequential use of letrozole and human menopausal gonadotrophin (HMG) was compared with HMG only in poor ovarian responders undergoing IVF. Patients (n=53) with less than four oocytes retrieved in previous IVF cycles or less than five antral follicles were randomized to either letrozole for 5days followed by HMG or HMG alone. The letrozole group had lower dosage of HMG (P<0.001), shorter duration of HMG (P<0.001) and fewer oocytes (P=0.001) when compared with controls. Live-birth rate was comparable with a lower miscarriage rate in the letrozole group (P=0.038). Serum FSH concentrations were comparable in both groups except on day 8, while oestradiol concentrations were all lower in the letrozole group from day <em>4</em> (all P<0.001). Follicular fluid concentrations of testosterone, <em>androstenedione</em>, FSH and anti-Müllerian hormone were higher in the letrozole group (P=0.009, P=0.001, P=0.0<em>4</em>6 and P=0.03<em>4</em>, respectively). Compared with HMG alone, sequential use of letrozole and HMG in poor responders resulted in significantly lower total dosage and shorter duration of HMG, a comparable live-birth rate, a significantly lower miscarriage rate and a more favourable hormonal environment of follicular fluid. The management of poor ovarian responders or women with poor ovarian reserve in IVF is controversial. The use of letrozole has been studied; however, results are inconsistent. This randomized trial studied the sequential use of letrozole and gonadotrophin compared with gonadotrophin alone in poor responders undergoing IVF. The sequential use of letrozole and gonadotrophin led to a significantly lower dosage and shorter duration of gonadotrophin use, significantly fewer oocytes, comparable live-birth rate, a significantly lower miscarriage rate and a more favourable hormonal environment at a lower cost.

To investigate the influence of obesity on hormonal parameters in 186 apparently healthy women and in 176 women suffering from severe obesity the serum concentrations of FSH, LH, estrone (E1), estradiol (E2), <em>androstenedione</em> (A) and testosterone (T) were determined radioimmunologically. The climacteric onset of increased FSH production is <em>4</em> yr earlier (P less than 0.001) in obese than in normal women. Parallel to the rise of FSH there is a significantly premature decrease of the E1 and E2 concentrations in obese women. The typical elevation of the LH was found similar and not significantly different in the two collectives. The mean A levels are significantly lower (P less than 0.01) in obese than in normal women in all age groups. The T concentrations do not depend on the age of the women during the investigated period (<em>4</em>1 to 60 yr) and are significantly higher (P less than 0.001) in the obese than in the normal women. There is a significant (P less than 0.001) correlation between the concentrations of A and E1 both in the obese and the normal women. An increased conversion of androgens to estrogens by adipose tissue is not revealed by the peripheral serum concentrations. Our data clearly demonstrate that in obese women the onset of ovarian insufficiency is significantly earlier than in normal women.

Malnutrition in HIV-infected patients is characterized by a loss of both fat-free mass (FFM) and fat mass (FM). Glucocorticoids and androgens change during the course of the infection and may play a key role in the protein balance. The serum concentrations of cortisol, adrenal (DHEA and DHEA sulfate) and gonadal androgens (<em>androstenedione</em>, testosterone, and dihydrotestosterone) of HIV-positive men were measured and compared with several parameters of body composition as a function of body weight loss (BWL). The patients were assigned to one of five groups according to their BWL: group I (controls, n = 10) < 5%, group II (n = 7) 5-10%, group III (n = 8) 10.1-16%, group IV (n = 9) 16.1-2<em>4</em>%, and group V (n = <em>4</em>)>> 2<em>4</em>.1%. Correlation analysis showed significant positive or negative relationships between several markers of malnutrition and adrenal androgens and the cortisol:DHEA ratio, but not with cortisol. BWL was negatively correlated with DHEA (r = -0.69, P < 0.0001), DHEA sulfate (r = -0.58, P < 0.0001) and testosterone (r = -0.3<em>4</em>, P < 0.03), but positively with the cortisol:DHEA ratio (r = 0.61, P < 0.0001). In contrast, BCM was positively correlated with DHEA (r = 0.3<em>4</em>, P < 0.0<em>4</em>) and DHEA sulfate (r = 0.36, P < 0.03) and negatively with the cortisol:DHEA ratio (r = -0.58, P < 0.0001). The cortisol:DHEA ratio was also negatively correlated with BMI (body mass index) (r = -0.56, P < 0.01), fat-free mass (r = -0.<em>4</em>8, P < 0.00<em>4</em>), fat mass (r = -0.39, P < 0.02), and BCM:weight ratio (r = -0.<em>4</em>7, P < 0.005) and positively with the extracellular:intracellular water ratio (r = 0.5<em>4</em>, P < 0.001). These data indicate that the steroid hormone environment of patients, particularly their cortisol:DHEA ratio, is linked to the malnutrition associated with HIV infection. The decreased DHEA and increased cortisol in patients with the advanced stages of disease could be associated with increased protein catabolism.

In a case-control study nested within the Greek EPIC cohort, serum levels of estrone, estradiol, <em>androstenedione</em>, dehydroepiandosterone sulfate, testosterone and IGF-1 were measured for 29 breast cancer patients and 58 control women, matched for age and menopausal status. There was little difference in breast cancer risk when values of <em>4</em>-6-as compared to values of 1-3-hormones were elevated, a finding arguing against a positive interaction among these hormones. Breast cancer risk, however, was significantly and substantially lower among women with levels of all hormones below the corresponding age- and menopausal-status-predicted means, compared to women with levels of at least 1 hormone above the predicted mean (odds ratio = 0.11 with 95% confidence interval 0.01-0.90; p = 0.0<em>4</em>). Our results suggest that the studied mammotropic hormones may act as permissive factors for breast cancer occurrence, and that the levels of some of them above the mean suffice for sustaining growth of a developing tumor. A corollary is that studies of mammotropic hormones in relation to breast cancer risk may also need to focus on the lower end of the distributions of these growth-enhancing hormones.

Steroidogenic factor 1 (SF-1) is a master regulator for steroidogenesis. In this study, we identified novel SF-1 target genes using a genome-wide promoter tiling array and a DNA microarray. SF-1 was found to regulate human glutathione S-transferase A (GSTA) family genes (hGSTA1-hGSTA<em>4</em>), a superfamily of detoxification enzymes clustered on chromosome 6p12. All hGSTA genes were up-regulated by transduction of SF-1 into human mesenchymal stem cells, while knockdown of endogenous SF-1 in H295R cells down-regulated all hGSTA genes. Chromatin immunoprecipitation assays, however, revealed that SF-1 bound directly to the promoters of hGSTA3 and weakly of hGSTA<em>4</em>. Chromosome conformation capture assays revealed that the coordinated expression of the genes was based on changes in higher-order chromatin structure triggered by SF-1, which enables the formation of long-range interactions, at least between hGSTA1 and hGSTA3 gene promoters. In steroidogenesis, dehydrogenation of the 3-hydroxy group and subsequent Δ(5)-Δ(<em>4</em>) isomerization are thought to be enzymatic properties of 3β-hydroxysteroid dehydrogenase (3β-HSD). Here, we demonstrated that, in steroidogenic cells, the hGSTA1 and hGSTA3 gene products catalyze Δ(5)-Δ(<em>4</em>) isomerization in a coordinated fashion with 3β-HSD II to produce progesterone or Δ(<em>4</em>)-<em>androstenedione</em> from their Δ(5)-precursors. Thus, hGSTA1 and hGSTA3 gene products are new members of steroidogenesis working as Δ(5)-Δ(<em>4</em>) isomerases.

OBJECTIVE

To evaluate the systemic and therapeutic effect of topical testosterone treatment in vulvar lichen sclerosus.

METHODS

This prospective clinical, single-arm study included ten postmenopausal women with vulvar lichen sclerosus. Testosterone propionate (0.0<em>4</em> g daily) was administered topically for <em>4</em> weeks. Serum androgens (testosterone, free testosterone, <em>androstenedione</em>, dehydroepiandrosterone sulfate) were determined before and after <em>4</em> weeks of treatment, and vulvodynia was evaluated by a horizontal visual analogue scale.

RESULTS

Serum levels of total testosterone increased in all patients (P < .01) and exceeded normal range in eight of ten women. Vulvodynia improved in nine of ten patients (paired t test: P < .01). Four of ten patients showed clinical signs of hyperandrogenism (enlargement of the clitoris, alterations of the voice, increase in libido) after <em>4</em> weeks of treatment. The only patient without subjective improvement had elevated basal serum androgen levels and showed clinical signs of hyperandrogenism before therapy.

CONCLUSIONS

Topical testosterone is effective in normoandrogenic women with lichen sclerosus. Androgen status should be evaluated before treatment, and dosage should be individualized to avoid virilization and metabolic side effects. Because there is a marked systemic effect, clinical controls and a follow-up with evaluation of serum testosterone levels are recommended. Other steroids should be included in therapeutic decisions.

The granulosa cell produces an inhibitor of aromatase activity, which recently was purified to homogeneity (follicle-regulatory protein: FRP). Since extracts of testicular homogenates also contain factor(s) with biological properties similar to FRP, including inhibition of granulosa cell aromatase, we examined the effects of ovarian FRP on testicular function. Forty-five-day-old rats received daily FRP injections (100 micrograms or 300 micrograms). After 15, 30, <em>4</em>5, and 70 days of therapy, (n = 5 each group), trunk serum was measured for testosterone, <em>androstenedione</em>, estradiol, and FSH levels by established radioimmunoassays (RIA). One testis from each rat was homogenized, centrifuged, and evaluated for sperm head counts; the other testis was dissected by transillumination, and the length of seminiferous epithelial stages determined. After 15 (control: <em>4</em>.8 +/- 0.2 mm; 100 micrograms: 6.0 +/- 0.3 mm; 300 micrograms: 6.6 +/- 0.3 mm) and 30 days (control: <em>4</em>.6 +/- 0.2 mm; 100 micrograms: 6.3 +/- 0.2 mm; 300 micrograms: 5.9 +/- 0.2 mm) of treatment the length of the "strong" seminiferous tubule segment in FRP-treated rats was greater than in control rats (p less than 0.05). Serum levels of steroids and FSH were similar in all groups. After 30, <em>4</em>5, and 70 days of treatment, the sperm head counts for the 100-micrograms and 300-micrograms dosages were 26%, 29%, 30% and 20%, 3<em>4</em>%, and 2<em>4</em>% of control values, respectively. By 70 days of treatment, cycle Stage VII was markedly reduced or absent in FRP-treated rats, and their round spermatids contained ring chromatin; both conditions indicate degeneration. FRP (50 micrograms/ml) was added to rat Sertoli cell cultures for <em>4</em> days after which transferrin and androgen-binding protein (ABP) were measured. FRP enhanced Sertoli cell secretion of ABP (58 vs. 138 +/- 7 microliters eq/culture) and transferrin (2.1 vs. <em>4</em>.7 +/- 0.6 microgram/culture). In conclusion, systemic injection of FRP alters seminiferous epithelial function by reducing maturation of mature sperm forms. Adding FRP to Sertoli cells in culture enhances secretion of transferrin and ABP; this suggests that maturation of the germinal elements may be linked to the secretory function of seminiferous tubules.

A hallmark of severe congenital adrenal hyperplasia due to 21-hydroxylase deficiency is pre- and postnatal virilization. The most characteristic biochemical abnormality is the elevation of 17α-hydroxyprogesterone, which is metabolized to the most potent androgen receptor agonist dihydrotestosterone. 17α-Hydroxyprogesterone can be metabolized to dihydrotestosterone via <em>4</em>-<em>androstenedione</em> through the classical Δ⁴-pathway or via 17α-hydroxypregnenolone and dehydroepiandrosterone through the classical Δ⁵-pathway, as well as through an alternative route, called the 'backdoor pathway', that bypasses dehydroepiandrosterone, <em>4</em>-<em>androstenedione</em>, and testosterone as intermediates. This review article will summarize recent advances in the understanding of the activities of androgen synthesis pathways in patients with 21-hydroxylase deficiency obtained by urinary steroid metabolomics based on gas chromatography-mass spectrometry. Compared with healthy controls, the relative activities of the backdoor and Δ⁴-pathways increase in patients with congenital adrenal hyperplasia during neonatal age and infancy, whereas the activity of the Δ⁵-pathway remains unchanged. Thereafter, the activity of the Δ⁵-pathway dominates, whereas a decreasing 5α-reductase activity leads to a diminished role of the backdoor pathway for androgenic steroid production. Beside the backdoor pathway, the Δ⁴-pathway seems to be responsible for increased androgen generation in patients with 21-hydroxylase deficiency before the onset of adrenarche, whereas the Δ⁵-pathway might contribute to the increased androgen formation in those patients only after the onset of adrenarche.

We have tested the hypothesis that idiopathic hirsutism (IH) may be due to abnormality of androgen-responsive hair follicles. Because androgen metabolism within target cells is an important determinant of androgen action, we have analyzed the rates of formation and disposition of the major mediators of androgen action, testosterone (T) and dihydrotestosterone (DHT). In normal women, the pattern of androgen metabolism by growing hairs favors T predominance over DHT and inactivation of both these 17 beta-hydroxysteroids to 17-ketosteroids. This pattern results greatly from predominance of 17 beta-hydroxysteroid dehydrogenation. For example, in normal women's scalp hair, DHT disposition to 5 alpha-androstanedione proceeded at the rate of 8.6 +/- 2.0 (SEM) %/micrograms DNA/min, whereas DHT was formed from T at a rate of 0.1<em>4</em> +/- 0.02, and T was formed from <em>androstenedione</em> at a rate of 0.60 +/- 0.12, all significantly different from one another. Both the formation of 17-ketosteroids and the apparent 5 alpha-reductase activity were exaggerated in the pubic hair of men; whether these differences are site-, sex-, or androgen-related, remains to be determined. Pubic hairs tended to metabolize androgens at a greater rate than did scalp hair. This was related to the significantly greater DNA content of plucked pubic hairs, a difference unrelated to sex or androgen levels. Women with IH had heterogeneous pubic hair abnormalities. Only 1 of the <em>4</em> IH patients studied had abnormal pubic hair follicle androgen metabolism, with the greatest abnormality being an exaggerated rate of 17 beta-hydroxysteroid inactivation to 17-ketosteroids. Two of the other 3 IH cases had increased DNA content of plucked pubic hairs, a different kind of exaggeration of normal, which suggests an abnormality of hair follicle growth unrelated to androgen sensitivity. We favor the concept that IH is related to various distinct types of sexual hair abnormalities which reflect fundamental defects in the regulation of hair growth.