We report an 18-month-old girl with Cushing's disease caused by a large adenoma of the pituitary gland. Tumour size and extension were determined by X-ray, CT-scan and angiographic studies. The endocrinological findings were typical for this disease: elevated plasma levels of ACTH, cortisol, <em>17</em>-Hydroxyprogesterone (<em>17</em>-OHP) and testosterone, elevated urinary excretion of <em>17</em>-<em>Ketosteroids</em> (<em>17</em>-KS) and <em>17</em>-Hydroxycorticoids (<em>17</em>-OHCS). Dexamethasone failed to suppress ACTH and cortisol plasma levels. TRH induced only a minimal TSH increase. Following LH-RH injection gonadotropin levels rose to pubertal values. The hGH response to insulin-induced hypoglycaemia was subnormal. After resection of the tumour the infant died because of non-treatable arrhythmia. Histological findings showed a non-differentiated adenoma with extension into the subarachnoid space and into nerve tissues. In vitro lysine-vasopressin (LVP) and arginine-vasopressin (AVP) exhibited only weak stimulatory effects on the ACTH secretion of the tumour cells.

Serum acid phosphatase activity, urinary total cholesterol, and ratio of deoxy to oxy urinary <em>17</em>-<em>ketosteroids</em> were measured in a group of 42 patients with prostatic carcinoma and in a group of 14 age-matched normal healthy individuals. Our purpose was to evaluate whether or not the simultaneous determinations of these tests would increase the rate of detection obtained by the single assay alone. The results of single assay revealed for the following detection rate: 67 per cent (28 of 42 patients) for serum acid phosphatase, 62 per cent for urinary total cholesterol, and 22 per cent for ratio of <em>17</em>-<em>ketosteroids</em>. A significant increase of detection rate was observed when simultaneous determinations of two assays were performed; 86 per cent for serum acid phosphatase activity and total urinary cholesterol; 74 per cent for serum acid phosphatase and ratio of <em>17</em>-<em>ketosteroids</em>; and 74 per cent for total urinary cholesterol and ratio of <em>17</em>-<em>ketosteroids</em>. A detection rate of 88 per cent (37 of 42 patients) was obtained as all three assays were analyzed, though it was not significantly different from a ratio of 86 per cent for simultaneous assays of acid phosphatase and total cholesterol. It was concluded that simultaneous determinations of serum acid phosphatase activity, urinary total cholesterol, and androgens are of values in diagnosis for patients with prostatic neoplasia.

We report the clinical history and results of endocrine investigations in two brothers born to consanguineous parents, who presented with hypokalemia and arterial hypertension when they were aged 2 and 6 years. The hormonal serum assay results, including extremely low values for aldosterone and plasma renin activity, favored the existence of apparent mineralocorticoid excess. A diagnosis of 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD) deficiency was made, based on assays of the hydrogenated urinary metabolites of cortisol and cortisone, as well as of corticosterone and dehydrocorticosterone. Indeed we found a very low rate of urinary elimination of cortisone metabolites: tetrahydrogenated cortisone was reduced to between 0.10 and 30 mumol/24 h, which is 15-100 times lower than the normal rate; hexahydrogenated cortolones alpha and beta were found to be 7- to 20-fold lower than normal levels; and the 11-keto-<em>17</em>-<em>ketosteroid</em> derivatives of cortisone were also reduced. Urinary elimination of the cortisol-reduced metabolites 5 beta- and 5 alpha-tetrahydrogenated cortisol were slightly reduced or normal. These results argue in favor of a deficit in the enzyme 11 beta-HSD, which oxidizes cortisol into cortisone. A moderate defect in the conversion of cortisol into 5 beta-THF compared to normal conversion into 5 alpha-THF was also found. With respect to corticosterone metabolism, we demonstrated the presence of a defect in the oxidation of that steroid into dehydrocorticosterone, also due to the deficit in 11 beta-HSD. Arterial hypertension and hypokalemia were corrected by treatment with dexamethasone, concomitantly with correction of the low aldosterone and plasma renin activity levels. On the other hand, during this treatment, urinary concentrations of the metabolites of cortisol, cortisone and corticosterone were only moderately affected.

Many steroid hormones contain a Δ(4)-3-<em>ketosteroid</em> functionality that undergoes sequential reduction by 5α- or 5β- steroid reductases to produce 5α- or 5β-dihydrosteroids; and a subsequent 3-keto-reduction to produce a series of isomeric tetrahydrosteroids. Apart from steroid 5α-reductase all the remaining enzymes involved in the two step reduction process in humans belong to the aldo-keto reductase (AKR) superfamily. The enzymes involved in 3-<em>ketosteroid</em> reduction are AKR1C1-AKR1C4. These enzymes are promiscuous and also catalyze 20-keto- and <em>17</em>-keto-steroid reduction. Interest in these reactions exist since they regulate steroid hormone metabolism in the liver, and in steroid target tissues, they may regulate steroid hormone receptor occupancy. In addition many of the dihydrosteroids are not biologically inert. The same enzymes are also involved in the metabolism of synthetic steroids e.g., hormone replacement therapeutics, contraceptive agents and inhaled glucocorticoids, and may regulate drug efficacy at their cognate receptors. This article reviews these reactions and the structural basis for substrate diversity in AKR1C1-AKR1C4, <em>ketosteroid</em> reductases. This article is part of a Special Issue entitled 'Steroid/Sterol signaling'.

Theophylline is thought to improve asthma by increasing intracellular cyclic adenosine 3'-5'-monophosphate (cAMP) levels. It has been demonstrated in experimental animals that elevation of intracellular cAMP in the adrenal cortex causes an increased secretion of cortisol. We studied whether therapeutic doses of theophylline given intravenously and orally to human subjects over 3 days would increase cortisol secretion. A single-blind, 6-day protocol was employed in five normal and five asthmatic volunteers. Adrenal function was monitored by 8 A.M. and 4 P.M. serum cortisol and adrenocorticotropic hormone (ACTH) levels; daily 24-hr urine for urinary-free cortisol (UFF), <em>17</em>-hydroxysteroids (<em>17</em>-OH), and <em>17</em>-<em>ketosteroids</em> (<em>17</em>-KS); and alternate-day cortisol secretory rates (FSR) measured by isotope dilution after intravenous 14C-cortisol. Serum theophylline concentration also was monitored. Results in normal and asthmatic subjects were similar. Theophylline caused a significant but transient increase in UFF and <em>17</em>-OH excretion. Urine volumes also increased significantly, suggesting that the renal effect of theophylline accounted for the increased UFF and <em>17</em>-OH excretion. FSR increased during the first 24 hr after theophylline in eight of nine cases (p < 0.05 by sign test), mean values increasing from 14.2 to 19.3 mg, but this effect had dissipated by day 3 of theophylline administration. In contrast to these findings, theophylline had no effect on serum cortisol or ACTH or urinary <em>17</em>-KS. It is likely that serum cortisol and ACTH remained unchanged because the increase in cortisol secretion was offset by a concomitant increase in cortisol clearance. It is concluded that theophylline produces a small, transient increase in cortisol secretion and clearance, and this effect is similar in asthmatic and normal subjects.

To reinvestigate the effect of hCG on circulating and urinary dehydroepiandrosterone sulfate (DHEAS), a hCG stimulation test (5000 IU administered i.m. at 8.30 h on 3 consecutive days) was performed in 6 healthy males (aged 24 to 35 years). Blood specimens and 24-h urine samples were collected immediately before the first and directly after the last hCG administration. Contrary to previous findings in normal men, the present study revealed significant DHEAS responses after testicular stimulation with hCG: plasma DHEAS increased from 7.9 +/- 2.3 to 9.6 +/- 2.2 mumol/L (P < 0.05) and urinary DHEAS from 5.7 +/- 3.6 to 9.3 +/- 5.2 mumol/day (P < 0.05). There was also a marked rise (P < 0.05) in the urinary excretion of total <em>17</em>-<em>ketosteroid</em> sulfates. Clear increases of unconjugated plasma dehydroepiandrosterone as well as of circulating and renally excreted androstenedione and testosterone definitely confirmed an adequate Leydig cell stimulation. Significant post-hCG changes were additionally observed for plasma and urinary 3 alpha-androstanediol glucuronide (149% and 79% increases, respectively) and for urinary cortisol (21% decrease). Significant correlations were found for the post-hCG percent increases of plasma androstenedione versus plasma DHEAS (r = 0.86) and for the percent increases of plasma testosterone versus urinary DHEAS (r = 0.98), indicating that the extent of gonadal androgen elevations in the circulation of normal men is a determinant of DHEAS increases in blood or urine. These findings provide an explanation for the frequently observed sex differences for DHEAS in adults.(ABSTRACT TRUNCATED AT 250 WORDS)

During an altitude training camp (23 days, 1850 m above sea level) we collected night urine from 36 German elite rowers in order to measure the excretion of urea, creatinine, <em>17</em>-<em>ketosteroids</em> (<em>17</em>-KS), and <em>17</em>-hydroxycorticosteroids (<em>17</em>-OHCS). <em>17</em>-KS and <em>17</em>-OHCS represent the major metabolites from endogenous anabolic and catabolic steroid hormone systems. There was no significant change in the excretion of <em>17</em>-OHCS during the training camp (mean value: females 0.0025 +/- 0.0011, males 0.0033 +/- 0.0012 mg/kgbw.h). Significant increases in the excretion of <em>17</em>-KS (mean value: females 0.007 +/- 0.003, males 0.0092 +/- 0.0039 mg/kgbw.h) and in the ratio <em>17</em>-KS/<em>17</em>-OHCS (mean value: females 3.13 +/- 1.65, males 3.11 +/- 1.71) were found during the first week and towards the end of the training camp. To investigate training effects on the excretion of these hormone metabolites, we used a recently described classification for rowing training (categories I to IV of rowing-specific training according to lactate levels and non-rowing specific categories) and a corresponding rowing data base. More than 80% of training was performed at a lactate level lower than 2 mmol/l. Using multiple regression analysis, the general finding was that in males the ratio of <em>17</em>-KS/<em>17</em>-OHCS increased with rowing specific and non specific training regimes where a lactate level below 2 mmol/l was observed. Training at higher lactate levels caused a decrease in the ratio that may be interpreted as a shift in the production from endogenous androgenic steroid hormones to cortisol. No significant effects of single training variables were found in female rowers, which indicates major training influences on testosterone metabolism. The influence of further factors such as a relationship between urine urea and <em>17</em>-KS in males are described and need further explanation.

The measurement of androgen steroids has been utilized as a clinical indicator of adrenal function, androgen abuse, and as a prediction of general health or biological aging. An improved high-performance liquid chromatography-ion trap mass spectroscopic method with sonic spray ionization (SSI) technology for the quantification of individual urinary <em>17</em>-<em>ketosteroid</em> sulfates and glucuronides was developed and validated. Sample preparation was simplified using a C18 cartridge followed by direct injection onto a reversed-phase HPLC column. Individual <em>17</em>-<em>ketosteroid</em> from 63 urinary specimens collected in a 24-h period was measured. <em>17</em>-<em>Ketosteroid</em> conjugates, total <em>17</em>-KS-S and the ratio of total <em>17</em>-KS-S to creatinine referred to herein as the Anabolic/Catabolic Index (ACI) showed statistically significant negative correlations with age.

Although androgens have been implicated in follicular atresia, ovarian follicular androgen synthesis is required for preovulatory follicular growth. To localize the site(s) of androgen biosynthesis and to obtain a better understanding of the regulation of the androgenic pathway(s) in rat ovarian follicles we examined the relative abilities of developing follicles to accumulate specific androgens [testosterone (T) and dihydrotestosterone (DHT)] using both radioimmunoassay (RIA) and 3H-substrate metabolism techniques. Small antral and preovulatory follicles were obtained from control or human chorionic gonadotropin (hCG)-primed immature rats, respectively (Richards and Bogovich, 1982). Small antral follicles, theca and granulosa cells produced little immunoassayable androgen (T + DHT) when incubated with or without 8-bromo-cAMP. In contrast, preovulatory follicles and theca produced more androgen than small antral tissues and in a manner acutely stimulable by cAMP. Granulosa cells produced little androgen under these conditions. Inclusion of [3H] androstenedione in the incubates yielded increased accumulation of [3H] T and [3H] DHT for all small antral and preovulatory tissues. Indeed, granulosa cells from both small antral and preovulatory follicles possessed a remarkable ability to accumulate [3H] T. This ability was not altered by hypophysectomy or subsequent treatment with estradiol and/or follicle-stimulating hormone (FSH). These results suggest that <em>17</em>-<em>ketosteroid</em> reductase may be a constitutive enzyme in granulosa cells.

Influence of the competitive testosterone antagonist 4-nitro-3-trifluoromethylisobutyranilide (NFBA) on the hypothalamo-hypophyseal-gonadal system in male Wistar rats and guinea pigs was studied. NFBA as well as nonlabelled testosterone diminished 3H-testosterone uptake in hypothalamus and hypophysis of castrated animals. Increase of plasma testosterone level was obtained after the antiandrogen treatment in a dose of 10 or 25 mg/kg per os during 3, 10 and 48 days. Maximal level of the hormone was noticed on the 10th day. The contents of <em>17</em>-<em>ketosteroids</em> in testes had not changed, but the testosterone content and steroid-delta5-3beta-ol-dehydrogenase activity were high during the treatment period. After 10 days of NFBA administration increased plasma LH level was found. Increase in size and number of gonadotropocytes was revealed by histological and histochemical methods as well as Leydig cell hypertrophy. High plasma testosterone level and Leydig cell activation were obtained in guinea pigs as a result of NFBA subcutaneous injections in a dose of 10 mg/kg during 30 days. The suggestion is made that prolonged NFBA treatment produces changes in the testosterone metabolism. Short-term administration of NFBA may be used for the estimation of the hypothalamo-hypophyseal functional abilities.

We examined the serum concentrations of delta(5)-3beta-hydroxysteroids, pregnenolone (Preg), <em>17</em>-hydroxypregnenolone (<em>17</em>-OH-Preg), dehydroepiandrosterone (DHEA), androstenediol (ADIOL) and their sulfates in 30 well controlled (Group I: HbA1c<7.0%) and 15 poorly controlled (Group II: HbA1c>7.1%) type 2 diabetic patients, and 30 normal controls. These patients were treated with diet therapy or anti-diabetic agent. The distribution of gender and age of the subjects were matched between the groups. The serum levels of sulfo-conjugated and unconjugated steroids described above were measured by GC-MS and enzyme immunoassay (EIA), respectively. The serum levels of the entire sulfo-conjugated steroid measured in this study were significantly lower in Groups I and II than in controls. On the other hand, Preg levels in both Groups I and II were significantly higher than those in controls, whereas the serum levels of the downstream unconjugated steroids were not different from controls. To investigate the effect of sulfonylurea (SU) on the serum levels of steroids, the serum concentrations of steroids between the patients who were treated with diet therapy and SU agent were compared in Group I. No significant differences were observed between both groups. These results suggest that (1) since increased Preg levels did not cause any changes in the downstream delta(5)-3beta-hydroxysteroid levels, the metabolic pathway of delta(4)-3-<em>ketosteroids</em> may be accelerated in type 2 diabetes; (2) serum steroid levels were not affected by SU treatment; (3) sulfo-conjugated steroid catabolism was altered in type 2 diabetes; (4) the decreased sulfo-conjugated steroids especially ADIOLS may contribute to the alteration of sex steroid levels and onset or exacerbate infectious diseases in diabetes.

Feeding 50% ethanolic extract of A. aspera to male rats resulted in reduced sperm counts, weight of epididymis, serum level of testosterone and testicular activity of 3beta-hydroxysteroid dehydrogenase, while motility of the sperm and activity of the HMG CoA reductase were not affected. Cholesterol level in the testis, incorporation of labelled acetate into cholesterol, <em>17</em>-<em>ketosteroids</em> in urine and hepatic and fecal bile acids were increased. The results suggest that ethanolic extract of A. aspera caused reproductive toxicity in male rats and the action may be by suppressing the synthesis of androgen.

Bilateral slipped upper femoral epiphysis is a rare manifestation of renal osteodystrophy. A case of bilateral slipped femoral epiphysis in an 18-year old male suffering from chronic renal failure due to oligomeganephronic renal hypoplasia with profound signs of renal osteodystrophy is presented. Serum growth hormone levels were high, while those of urinary <em>17</em>-<em>ketosteroids</em> were decreased. Following subtotal parathyroidectomy, the progression of the process leading to slipped epiphysis was halted with closure of the epiphyses. The patient was subsequently treated with chronic hemodialysis for several months, after which successful renal transplantation was performed. The pathogenesis of renal osteodystrophy leading to slipped epiphysis is discussed and attention drawn to the fact that bilateral slipped femoral epiphysis may be the first clinical sign of chronic renal insufficiency.

Hydroxysteroid (<em>17</em>beta) dehydrogenase 1 (HSD<em>17</em>B1) catalyzes the reaction between the low active <em>17</em>-<em>ketosteroids</em> and the highly active <em>17</em>beta-hydroxysteroids. In the present study, we have generated transgenic (TG) mice expressing human (h) HSD<em>17</em>B1 under mouse mammary tumor virus (MMTV) promoter (MMTV-hHSD<em>17</em>B1TG mice). The MMTV-hHSD<em>17</em>B1TG mice were used to characterize HSD<em>17</em>B1 enzyme activity and properties of HSD<em>17</em>B1 inhibitor in vivo. Expression of the transgene was detected by enzyme activity and RT-PCR analysis. Increased HSD<em>17</em>B1 activity in the TG mice was detected in vivo by applying estrone as a substrate via an intravenous injection. The developed enzyme activity measurement was then applied to analyze the efficacy of HSD<em>17</em>B1 inhibitor in vivo. The results indicated that the MMTV-hHSD<em>17</em>B1TG mouse model is a valuable novel tool to test human HSD<em>17</em>B1 inhibition by various compounds in vivo. With the potent hHSD<em>17</em>B1 inhibitor compound tested, at highest an 85% and 33% inhibition of the enzyme activity in males and in females, respectively, was observed.

Evidence for the role of androgens in the male aggressive and sexual behavior is reviewed. Medroxyprogesterone acetate (MPA; Provera, Upjohn) has a marked antiandrogen property; it is effective in lowering the testosterone level and controlling certain otherwise intractable sex deviations. The finding in 6 patients treated for sex deviation are summarized. The effects of MPA in the treatment of 11 temporal lobe epileptics and 5 other patients with severe angry-aggressive behavior disorder are reported. Most temporal lobe epileptics responded well to MPA. Weight gain and earlier sleep were consistent side effects. The values of plasma testosterone, serum luteinizing hormone, and urinary <em>17</em>-<em>ketosteroids</em> were decreased by the treatment. Four patients were XYY individuals with lack of control over their sexual-aggressive or angry-aggressive impulses.

Clostridium scindens is a gut microbe capable of removing the side-chain of cortisol, forming 11β-hydro-xyandrostenedione. A cortisol-inducible operon (desABCD) was previously identified in C. scindens ATCC 35704 by RNA-Seq. The desC gene was shown to encode a cortisol 20α-hydroxysteroid dehydrogenase (20α-HSDH). The desD encodes a protein annotated as a member of the major facilitator family, predicted to function as a cortisol transporter. The desA and desB genes are annotated as N-terminal and C-terminal transketolases, respectively. We hypothesized that the DesAB forms a complex and has steroid-<em>17</em>,20-desmolase activity. We cloned the desA and desB genes from C. scindens ATCC 35704 in pETDuet for overexpression in Escherichia coli The purified recombinant DesAB was determined to be a 142 ± 5.4 kDa heterotetramer. We developed an enzyme-linked continuous spectrophotometric assay to quantify steroid-<em>17</em>,20-desmolase. This was achieved by coupling DesAB-dependent formation of 11β-hydroxyandrostenedione with the NADPH-dependent reduction of the steroid <em>17</em>-keto group by a recombinant <em>17</em>β-HSDH from the filamentous fungus, Cochliobolus lunatus The pH optimum for the coupled assay was 7.0 and kinetic constants using cortisol as substrate were Km of 4.96 ± 0.57 µM and kcat of 0.87 ± 0.076 min-1 Substrate-specificity studies revealed that rDesAB recognized substrates regardless of 11β-hydroxylation, but had an absolute requirement for <em>17</em>,21-dihydroxy 20-<em>ketosteroids</em>.

Microbiological synthesis of 7α- and 7β-hydroxy derivatives of testololactone and testolactone was developed based on bioconversion of dehydroepiandrosterone (DHEA) by fungus of Isaria fumosorosea VKM F-881 with subsequent modification of the obtained stereoisomers by actinobacteria. The first stage included obtaining of the stereoisomers of 3β,7(α/β)-dihydroxy-<em>17</em>a-oxa-D-homo-androst-5-en-<em>17</em>-ones in the preparative amounts. Then the conversion of 7-hydroxylated D-lactones obtained by selected actinobacteria of Nocardioides simplex VKM Ac-2033D, Saccharopolyspora hirsuta VKM Ac-666, and Streptomyces parvulus MTOC Ac-21v was studied. Under the transformation of 3β,7α-dihydroxy-<em>17</em>a-oxa-D-homo-androst-5-en-<em>17</em>-one and its corresponding 7β-stereoisomer by N. simplex VKM Ac-2033D and S. hirsuta VKM Ac-666 the 7α- and 7β-hydroxy-<em>17</em>a-oxa-D-homo-androst-4-ene-3,<em>17</em>-dione (7α- and 7β-hydroxytestololactone), 7α- and 7β-hydroxy-<em>17</em>a-oxa-D-homo-androsta-1,4-diene-3,<em>17</em>-dione (7α- and 7β-hydroxytestolactone) were obtained with molar yields in a range of 60.3-90.9 mol%. The crystalline products of 7α-hydroxytestololactone, 7α-hydroxytestolactone, and their corresponding 7β-hydroxy stereoisomers were isolated, and their structures were confirmed by mass spectrometry and ¹H-NMR spectroscopy analyses. The strain of Str. parvulus MTOC Ac-21v transformed 3β,7(α/β)-dihydroxy-<em>17</em>a-oxa-D-homo-androst-5-en-<em>17</em>-ones into the corresponding 3-keto-4-ene analogs and did not show 3-<em>ketosteroid</em> 1(2)-dehydrogenase activity. The activity of actinobacteria towards steroid D-lactones was hitherto unreported.The results contribute to the knowledge of metabolic versatility of actinobacteria capable of transforming steroid substrates and may be applied in the synthesis of potential aromatase inhibitors.