A rare case of an adrenal adenoma that produced virilization by an isolated secretion of testosterone without a concomitant increase in <em>17</em>-<em>ketosteroids</em> is reported. It was erroneously concluded that the patient's hypertestosteronemia was due to an ovarian disorder because the urinary <em>17</em>-<em>ketosteroids</em> were normal. It then was speculated that the adenoma's enzyme system was so efficient it was capable of converting the more prevalent androstenedione directly to testosterone without metabolizing it to <em>17</em>-<em>ketosteroids</em>. Removal of the adrenal adenoma completely cured the patient's hirsutism.

<em>17</em>β-Hydroxysteroid dehydrogenase/<em>17</em>-<em>ketosteroid</em> reductase (<em>17</em>HSD/KSR) activity and <em>17</em>HSD/KSR types 1, 2, 4, and 5 mRNA levels were characterized in ovarian cancer cell lines derived from patients unexposed to radiation or chemotherapy. Activity was at the limit of detection in TOV-112D and TOV-21G cells. Activity in OV-90 was comparable to that in human placental tissue, was predominantly microsomal and was <em>17</em>HSD/KSR type 2-like in substrate specificity and inhibition patterns. In monolayers, conversion of testosterone (T) to androstenedione (A) was 12-fold greater than that of A to T. Reduction of fetal bovine serum to 0.3% in the culture medium had no effect on <em>17</em>β-HSD activity. Significant levels of type 1 and type 2 mRNAs were observed in OV-90 while only trace amounts were detected in TOV-21G. In contrast, type 4 mRNA levels were comparable for OV-90 and TOV-21G. Type 5 mRNA was detected in both cell lines but its level in OV-90 was twice that of TOV-21G. In OV-90, the type 2-like activity was predominant even though the type 5 mRNA level was 2.5-fold higher than that of the type 2. OV-90 cells may be a useful system for studying the regulation of <em>17</em>HSD/KSR type 2 activity and expression in ovarian epithelial cancer.

BACKGROUND

<em>17</em>-<em>Ketosteroid</em> reductase deficiency results in male pseudohermaphroditism because conversion of the weak androgen androstenedione to the more potent androgen testosterone is impaired. If a late-onset form exists, hypogonadism and gynecomastia caused by decreased testosterone production and increased estrogen production, respectively, would be expected as the major clinical manifestations in men.

METHODS

We studied 48 male subjects, ranging from 14 to 26 years of age, who had idiopathic pubertal gynecomastia. Serum concentrations of gonadal and adrenal steroid hormones were measured before and after the administration of corticotropin and after the combined administration of chorionic gonadotropin and dexamethasone for three days.

RESULTS

We identified three unrelated subjects (ages, 16, <em>17</em>, and 26 years) with results indicative of a partial deficiency of testicular <em>17</em>-ketosteroid reductase. The three subjects had gynecomastia as well as decreased libido and impotence. Their mean (+/- SD) base-line serum androstenedione and estrone concentrations were elevated as compared with the levels in the 45 subjects without this enzyme deficiency (androstenedione, 380 +/- 70 vs. 110 +/- 70 ng per deciliter [13 +/- 2 vs. 4 +/- 2 nmol per liter]; estrone, 138 +/- 12 vs. 46 +/- 9 pg per milliliter [511 +/- 44 vs. <em>17</em>0 +/- 33 pmol per liter]). After the administration of chorionic gonadotropin, the mean serum androstenedione concentration in these three subjects was 910 +/- 48 ng per deciliter (32 +/- 2 nmol per liter) and the mean serum estrone concentration was 260 +/- 16 pg per milliliter (962 +/- 59 pmol per liter). The mean serum testosterone concentration at base line was 210 +/- 80 ng per deciliter (7.4 +/- 2.8 nmol per liter) in the 3 subjects, as compared with a value of 410 +/- 12 ng per deciliter (14.4 +/- 0.42 nmol per liter) in the 45 other subjects, and it did not increase in response to the administration of chorionic gonadotropin. The concentrations of androstenedione and estrone in spermatic venous serum were 19 times higher and 73 times higher, respectively, than in normal men. The serum concentrations of follicle-stimulating hormone and luteinizing hormone in these three subjects were inappropriately low, suggesting the presence of hypogonadotropic hypogonadism.

CONCLUSIONS

A late-onset form of testicular <em>17</em>-ketosteroid reductase deficiency can cause gynecomastia and hypogonadism in men.

The inducible 3-keto-5 alpha-steroid-delta 4-dehydrogenase of Nocardia corallina was purified to homogeneity using affinity chromatography on 19-nortestosterone-<em>17</em>-acetoxyaminoethyl Sepharose 4B. SDS-polyacrylamide gel electrophoresis, gel filtration and spectral analysis of flavin suggest that the purified dehydrogenase is a monomeric protein of Mr 60,000 containing one flavin. It has a typical absorption spectrum of flavoprotein with maxima at 457, 375, and 277 nm. The values shifted to 470 and 395 nm on binding of 19-nortestosterone. The enzyme catalyzed the dehydrogenation of 3-keto-5 alpha-steroid at the 4- and 5-position, e.g. the conversion of 5 alpha-androst-1-ene-3,<em>17</em>-dione to 1,4-androstadiene-3,<em>17</em>-dione with the reduction of phenazine methosulfate. The substrate 3-<em>ketosteroid</em> has essentially the 5 alpha-configuration. The enzyme did not reduce potassium ferricyanide but did reduce cytochrome c at a moderate rate, and exhibited only a weak steroid oxidase activity. Stereochemical study demonstrated that the enzyme abstracts the 4 beta, 5 alpha-hydrogens of the substrate as a hydrogen ion through a protein-based reaction and as a hydride ion by transfer to FAD, respectively. The enzyme oxidizes a wide variety of 3-keto-5 alpha-steroids but not 3 beta-hydroxysteroid. The dehydrogenase also catalyzed steroid transhydrogenation between 3-keto-5 alpha-steroid and 3-keto-1,4-diene-steroid. The properties of this enzyme are compared with those of 3-keto-steroid-delta 1-dehydrogenase.

delta 5-3-<em>Ketosteroid</em> isomerase from Pseudomonas putida is subject to photoinactivation by light of wavelengths greater than 300 nm, specifically in the presence of the competitive inhibitor, 1,4,6-androstatrien-3-one-<em>17</em> beta-ol (TEO). In the absence of this steroid or in the presence of the nonchromophoric steroidal competitive inhibitor, deoxycholate, the enzyme activity is essentially unaffected by irradiation. Deoxycholate protects the enzyme from the TEO-dependent reaction to a degree which is predictable from the concentrations of deoxycholate and TEO and their respective competitive inhibition constants, thus demonstrating that the inititial velocity of the photoinactivation is dependent upon the fraction of enzyme active sites occupied by TEO. Cholate, which is not a competitive inhibitor, does not protect enzyme activity. Amino acid analyses of hydrochloric acid hydrolysates of the photoinactivated enzyme show no significant differences from that of the native enzyme. However, the fraction of initial enzyme activity remaining correlates quantitatively with the disappearance of one of the four thiol groups in each polypeptide chain of the enzyme. Enzyme irradiated under the same conditions in the absence of TEO does not lose thiol groups.

3 beta, <em>17</em> beta-Hydroxysteroid dehydrogenase (3 beta <em>17</em> beta HSDH) is an NAD-dependent dehydrogenase which has a double specificity for the 3- and <em>17</em>-positions on the steroid skeleton. When dehydroepiandrosterone (DHEA) is used as steroid substrate, and the assay coupled with <em>ketosteroid</em>-isomerase, the two reactions occur alternately and each reaction on the 3-position produces a chromophoric molecule. These two reactions can follow one another without dissociation of the coenzyme from the enzyme binding site. This is confirmed by competition experiments with another dehydrogenase.

The peripheral levels of <em>17</em>-hydroxypregnenolone (<em>17</em>delta5P), progesterone (P), <em>17</em>-hydroxyprogesterone (<em>17</em>P), testosterone (T), 5alpha-dihydrotestosterone (DHT), androstenedione (A), androst-5-ene-3beta,<em>17</em>beta-diol (delts5diol), dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate (DHEA-S), estradiol-<em>17</em>beta (E2), and cortisol (F) were measured in peripheral and right adrenal venous blood of an amenorrheic patient with a right virilizing adrenal adenoma. Urinary <em>17</em>-<em>ketosteroids</em> were markedly elevated and were not suppressed on a low dose of dexamethasone (Dex) for 7 days. Peripheral T level was 1.2 ng/ml and DHEA-S was 13,500 ng/ml. Calculations of the ratios of adrenal venous gradients for delta5 and delta4 steroids suggest that the predominant pathway of steroid secretion used by the tumor was as follows: pregnenolone (delta5P) leads to <em>17</em>delta5P leads to DHEA leads to A leads to T. Following removal of the adenoma, T returned to normal levels but DHEA-S was still above normal at 4100 ng/ml. The patient became eumenorrheic with marked improvement at her hirsutism and virilization.

BACKGROUND

This study evaluates the metabolic and hormonal response in elite female gymnasts undergoing strenuous training and supplementation with SUPRO Brand Isolated Soy Protein. METHODS.

METHODS

14 top female gymnasts (Romanian Olympic Team), took part in a study to examine their hormonal metabolic profile and to investigate any possible changes resulting from a 4-month program of strenuous training and daily supplementation with soy protein at a level of 1 g/kg body weight. Gymnasts wtare randomly assigned to one of two groups seven to the Supplemented Group (A) and seven to the Non-Supplemented Group (B). Both groups took part in the same program, which consisted of strenuous training for 4-6 hours/day (except on Sunday, controlled food intake and supplements including vitamins and minerals. Group A received a supplement of Sports Beverage Protein Mix with SUPRO Isolated Soy Protein (ISP) twice daily. Group B received a placebo identical in appearance and flavour. Selected parameters were measured before and after the 4-month training program (lean body mass, fat mass, serum hemoglobin, protein, fats, urea and creatinine, liver enzymes, serum total calcium and magnesium, immunoglobulins, urinary mucoproteins, serum T3 and T4, estradiol, progesterone, prolactin, testosterone and urinary <em>17</em>-<em>ketosteroids</em>).

RESULTS

Results demonstrated that the Supplemented Group (A) had an increase in lean body mass and serum levels of prolactin (p < 0.01) and T4 and a decrease in serum alkaline phosphetases (p < 0.01). The Non-Supplemented Group (B) had a decreased level of serum T4 and an increased level of urinary mucoproteins (p < 0.05).

CONCLUSIONS

Our preliminary conclusions might suggest lower metabolic-hormonal stress in elite female gymnasts undergoing strenuous training and who received daily supplementation with isolated soy protein.

Impairment of cortisol metabolism, evidence of endogenous hypercortism and sensitivity to oral corticosteroids are known to occur in patients with chronic liver disease. Measurements were made of plasma 11-OHCS and urinary cortisol levels and other parameters of adrenal function in a group of such patients. The mean plasma total 11-OHCS was lower in patients than in control subjects, although this difference was not statistically significant and the normal circadian rhythm was maintained. However, in patients with chronic liver disease a greater proportion of the 11-OHCS was in the non-protein bound state resulting in an elevation of this fraction. This elevation of non-protein bound 11-OHCS must represent a resetting of the pituitary-adrenal feedback mechanism in these patients. Corticosteroid binding globulin was lower in patients than in control subjects, although the difference was not statistically significant. Urinary cortisol excretion was significantly reduced as was excretion of <em>17</em>-<em>ketosteroids</em>. Cortisol secretion rate was found to be normal. It is suggested that elevation of plasma non-protein bound 11-OHCS, resulting from impaired metabolism and reduced protein binding of cortisol in patients with hepatic disease, may explain the features of endogenous hypercorticism seen in such patients. Moreover, in the presence of impaired steroid metabolism and reduced protein binding, these patients may exhibit an increased sensitivity to corticosteroid therapy, and therefore administration of a reduced dosage may be advisable.

OBJECTIVE

To emphasize the potentially devastating consequences of topical application of corticosteroids.

METHODS

A case report is presented of a previously healthy 38-year-old woman in whom florid Cushing's syndrome developed after prolonged use of Lotrisone.

RESULTS

Four years of continuous topical administration of Lotrisone, a combination antifungal-corticosteroid product, led to the development of Cushing's syndrome. The cream had been applied to less than 10% of the total body surface area without use of occlusive dressings, and the patient had no evidence of liver dysfunction or systemic skin disorder. Laboratory studies revealed an unmeasurable corticotropin level, suppressed or unmeasurable cortisol levels, and suppressed or low-normal 24-hour urine collection values for <em>17</em>-hydroxy-steroids, <em>17</em>-<em>ketosteroids</em>, and urinary free cortisol, all of which confirm suppression of the hypothalamic-pituitary-adrenal axis.

CONCLUSIONS

(1) Betamethasone dipropionate can lead to the development of Cushing's syndrome, even in the absence of enhancing factors. (2) Combination medications can lead to the unintentional use of component agents. (3) Availability of potent corticosteroids as over-the-counter medications allows for inappropriate duration of therapy and inadequate medical supervision.

<em>Ketosteroids</em> (e.g., 19-nortestosterone) and phenolic steroids (e.g., <em>17</em> beta-estradiol and <em>17</em> beta-dihydroequilenin), which are potent competitive inhibitors of delta 5-3-<em>ketosteroid</em> isomerase (isomerase, EC 5.3.3.1) of Pseudomonas testosteroni, undergo significant polarization upon binding to the active site of the enzyme. The 10 nm red shift of the UV absorption maximum of the enone chromophore of 19-nortestosterone, which occurs in the enzyme-steroid complex, resembles that observed when this steroid is exposed to strong acid. The UV and fluorescence spectral changes of <em>17</em> beta-estradiol and <em>17</em> beta-dihydroequilenin in the enzyme-bound complex resemble the spectra of ionized phenolate species in aqueous basic solutions. Since most enzymes bind their substrates and competitive inhibitors in a solvent-inaccessible hydrophobic environment, and the generation of charges in such nonpolar environments is unfavorable, we investigated the possibility that the spectral perturbations of the steroids might arise from strong hydrogen bonding in nonpolar environments. For this purpose, the spectral properties of model compounds capable of forming intramolecular hydrogen bonds were studied in nonpolar solvents. Thus, 4-hydroxyandrost-4-ene-3,<em>17</em>-dione, in which the 4-hydroxyl group is intramolecularly hydrogen-bonded to the 3-carbonyl group through a five-membered ring, exhibits a lambda max of 276.0 nm, while the corresponding 4-methyl ether, 4-methoxyandrost-4-ene-3,<em>17</em>-dione, which cannot form an internal hydrogen bond, shows a lambda max of 258.5 nm in aqueous solution.(ABSTRACT TRUNCATED AT 250 WORDS)

An investigation was carried out to study the effect of mechanized agricultural work on the excretion of some corticosteroids and catecholamines in tractor-drivers during a seasonal field work. Group 1 (15 subjects) was examined three times: in the morning hours before starting work and at two intervals during the first and second half of the working day. In group two (11 subjects) the characteristics of the circadian rhythm for 24h were investigated in urine collected at six intervals. The results from both studies disclose a considerable increase of the total <em>17</em>-<em>ketosteroids</em>, total <em>17</em>-oxycorticosteroids, free 11-oxycorticosteroids, nor-adrenaline and adrenaline excretion levels which indicates overactivity of the hypothalamic-pituitary-adrenal and sympathetic nervous systems. The circadian rhythms of the examined parameters are characterized by an increasing of the mesors and a lowering of the amplitudes of the rhythmic variations. No significant changes of corticosteroids and catecholamines are established. Considerable interindividual differences in the course of excretion of the studies indices as well as the lack of marked circadian rhythm in about 50% of the subjects are found.

A 28 year old white women was found to have a cervical tumor in the 25th week of pregnancy. Pathologic examination revealed a nonkeratinizing small cell carcinoma. After delivery by cesarean section, pelvic lymph node exploration was carried out, and all 15 nodes were free of tumor. Her condition was staged as II-A, and she was treated with local radiation. Metastatic disease became manifest almost a year later and was histologically similar to her primary disease. A Cushingoid appearance was noticed and plasma cortisol levels were elevated. Twenty-four hour urinary <em>17</em>-hydroxycorticosteroid (<em>17</em>-OHCS) and <em>17</em>-<em>ketosteroid</em> (<em>17</em>-KS) levels were elevated and failed to suppress with dexamethasone. Plasma adrenocorticotropin (ACTH) level was elevated. Electron microscopic examination of the tumor tissue revealed neurosecretory granules. Immunoperoxidase stains for ACTH were positive. The patient's course was one of progressive decline and eventual death. A literature review revealed two other cases in which carcinoma of the uterine cervix was considered to be the source of ectopic ACTH. Some small cell carcinomas of the cervix may arise from cells of the APUD series. Small cell carcinoma of the uterine cervix may behave differently from the more commonly encountered keratinizing and large cell nonkeratinizing carcinomas of the cervix and may not respond as well to standard therapy. Ectopic hormone production, production of abnormal peptides or of vasoactive amines may be more common in small cell carcinoma of the cervix than is currently recognized, and these products may be clinically useful as tumor markers.