Activity and inhibition of 3-beta-hydroxysteroid dehydrogenase/delta-5-4-isomerase in human skin.

Activity and inhibition of 3 beta-hydroxysteroid dehydrogenase/delta 5-4-isomerase, a key example of biosynthesis of androgenic steroids, in human skin were studied. Whole-width dermal tissue specimens excised from various regions of the male and female body were investigated with an in vitro radioenzyme assay method using dehydroepiandrosterone as substrate. The Michaelis-Menten constant of the enzyme was found to be Km = 10nM and the maximal velocity was Vmax = 0.625 pmol produced 4-androstene-3,17-dione/mg protein/20 min. Activity of 3 beta-hydroxysteroid dehydrogenase/delta 5-4-isomerase in male inguinal skin (n = 8) was 0.132-0.412, in female abdominal skin (n = 4) 0.140-0.255, in perineal skin (n = 4) 0.138-0.962 pmol/mg protein/20 min. The synthetic steroids cyproterone acetate, 4-MA and epostane proved to be potent inhibitors, IC50 values were 150, 6.2 and 1.45 nM, respectively.

In vitro inhibition of testicular delta 5-3 beta-hydroxysteroid dehydrogenase and prostatic 5 alpha-reductase activities in rats and humans by strogen forte extract.

Clinical findings indicate that Strogen forte (a standardized extract of the plant Sabalis serrulata) can be use successfully in the medical treatment of benign prostatic hyperplasia. The aim of the present study was to investigate the possible inhibitory effects of the Strogen forte extract on rat and human testicular delta 5-3 beta-hydroxysteroid dehydrogenase (delta 5-3 beta-HSD) and prostatic 5 alpha-reductase (5 alpha-R). Strogen forte proved to be a direct inhibitor of medium effectivity, with similar IC50 values for rat testicular delta 5-3 beta-HSD (400 +/- 23 micrograms/ml) and human testicular delta 5-3 beta-HSD (212 +/- 8.6 micrograms/ml). 5 alpha-R activities were analysed by in vitro incubation of rat and human prostatic tissue homogenates with 14C-labelled testosterone as substrate in KRPG-DTT medium (pH = 7.4) with NADPH coenzyme, in air, at 37 degrees C, in the presence of Strogen forte extract. The results clearly demonstrate that Strogen forte is a potent inhibitor of prostatic 5 alpha-R, with IC50 values of 385 +/- 35.6 micrograms/ml for rat and 245 +/- 64.6 micrograms/ml for human prostatic 5 alpha-R. The present study has revealed that this plant extract inhibits not only prostatic 5 alpha-R, but also testicular delta 5-3 beta-HSD.

In vitro binding of 16-methylated C18 and C19 steroid derivatives to the androgen receptor.

The binding of androgens and structurally related analogues to the androgen receptor was studied. The in vitro experiments were carried out with cytosol of castrated rat prostate, using [3H]R1881 (methyltrienolone) as radioligand. The binding parameters measured were Kd = 1.25 x 10(-10) M and Bmax = 111 fmol (mg protein)-1. Ligand specificity was confirmed by competition experiments with known androgen, oestrogen and progestogen ligands. The receptor binding of substituted steroids was studied. The RBAs (relative binding affinities) of our recently synthetized 16-alkyl steroids were low. The only exception was the 17 beta-hydroxy-16 beta-methylestr-4-en-3-one, which exhibited the remarkable RBA of 22.9%.

In vitro inhibitory effects of 16-methyl-substituted steroids on 5 alpha-reductase in rat and human prostates.

The inhibitory effects (IC50) of 16-methyl steroids on 5 alpha-reductase were studied. The in vitro experiments were carried out with homogenates of rat and human prostates. The investigated 16-methyl steroids were found to be weak inhibitors. In comparison with the known 5 alpha-reductase inhibitor 4-MA (17 beta-N, N-diethylcarbamoyl-4-methyl-4-aza-5 alpha-androstan-3-one), the relative IC50 values of the studied compounds are 4.7 times or more greater than 4-MA in human prostate and 23.5 times or more greater than 4-MA in rat prostate. The IC50 values increase in the sequence 16 alpha-, 16 beta- and 16,16-dimethyl derivatives. In human prostate homogenates IC50 varies between 0.6 and 120, while in rat it ranges from 1.6 to 1000 microM. This shows that the enzyme of the human prostate is more sensitive than that of the rat prostate to the methyl-substituted compounds.

[In vitro inhibitory effects of 16-methyl-substituted steroids on 5 alpha-reductase in rat and human prostates]. / 16-metil-szubsztituált szteroidok in vitro gátló hatása az 5 alpha-reduktáz aktivitására patkány- és emberi prosztatában.

The inhibitory effects (IC50) of 16-methyl steroids on 5 alpha-reductase were studied. The in vitro experiments were carried out with homogenates of rat and human prostates. The investigated 16-methyl steroids were found to be weak inhibitors. In comparison with the known 5 alpha-reductase inhibitor 4-MA, the relative IC50 values of the studied compounds are 4.7 times or more greater than 4-MA in human prostate and 23.5 times or more greater than 4-MA in rat prostate. The IC50 values increase in the sequence 16 alpha, 16-beta- and 16,16-dimethyl derivatives. In human prostate homogenates IC50 varies between 0.6 and 120, while in rat it ranges from 1.6 to 1000 microM. This shows that the enzyme of the human prostate is more sensitive than that of the rat prostate to the methyl-substituted compounds. Acylation of the 17-hydroxy group significantly increases the IC50 values (cf. 8,11: from 4.8 to 23.5 and from 0.52 to 0.62;9,12: from 26.0 to 170.0 and from 0.58 to 1.4;15,17: from 3.9 to 35.0; and 16,18: from 5.6 to 58.0 microM). Whereas lack of a 19-CH3 group improves the inhibitory effect in the 16-unsubstituted compounds (1,19;14,22), the reverse hold in the 16-methylated derivatives (9,20; 10,21; 15,23; 16,24).

[Analysis of steroids and binding proteins in the blood of women with androgenetic alopecia]. / Szteroidok és kötöfehérjék vizsgálata androgen alopeciás nök szérumában.

Specific and sensitive radioimmune methods were used to study the steroid levels and binding capacities of their binding proteins in the serum of 30 women with androgenetic alopecia, in order to establish the proportions of the patients in whom hyperandrogenism or hypercorticism can be detected. It was found that the serum testosterone, dehydroepiandrosterone sulphate and cortisol levels did not differ from those for healthy women except dehydroepiandrosterone which was high. In 5 cases (16.6%), the binding capacity of the "sex hormone-binding globulin" was pathologically low, while in 6 patients (20%) the "free androgen index" was elevated. The binding capacity of the "corticosteroide-binding globulin" was pathologically low in 4 of 21 cases (19%) while the "free cortisol index" was high in 5 of these 21 patients (23.8%). Diane treatment (2 mg cyproterone acetate and 50 micrograms ethinyl-oestradiol)--was administered during 2-5 cycles (8 cases), the pathological dehydroepiandrosterone and dehydroepiandrosterone sulphate levels normalized, the testosterone level decreased to the lower limit of the normal range, the binding capacity of the "sex hormone-binding globulin" increased considerably, and the "free androgen index" fell below the normal range. Diane treatment, an essential improvement was observed in the condition of the patients: the hair became less greasy and its tendency to fall out was likewise moderated.

Adenomatous leydig cell hyperplasia of the testicle remnant after subtotal orchidectomy in the rat.

Subtotal orchidectomy is a suitable method to induce adenomatous Leydig cell hyperplasia in the testicular remnant without irradiation or toxic chemicals. Very expressed hyperplasia of the smooth endoplasmic reticulum was detectable electronmicroscopically in the newly formed Leydig cells. The activity of Δ5-3ß-hydroxysteroid dehydrogenase in the homogenate of the testicular remnant was found considerably elevated. These observations suggest a steroidogenetic activity of the proliferating Leydig cells.

Improvement of the reproductive performance of sows by treatment with a GnRH superactive analogue.

The effect of a Hungarian-made superactive analogue of GnRH (Ovurelin, D-Phe6-GnRH-EA, Reanal, Hungary) on the postpartal sexual function of sows was monitored. GnRH treatment was carried out on day 19 before weaning. The sows were inseminated at the first oestrus after weaning. GnRH treatment markedly increased litter size at weaning, substantially reduced (to 25 and 50%, respectively) the number of sows failing to come into oestrus after weaning, and increased the number of sows coming into oestrus within one week after weaning by 42.5% and 9%, respectively. These beneficial effects were particularly apparent on the far using closed management technology.

Simultaneous occurrence of selective ACTH insensitivity, achalasia and alacrimia accompanied by hyperlipoproteinaemia.

An extremely rare clinical syndrome on a 7-year-old-girl is presented. Besides isolated glucocorticoid insufficiency, achalasia and alacrima disturbance of the lipid metabolism was also detected--being a special feature of this case. The details of the endocrine workup is discussed, providing clues for the possible pathomechanism. The correct diagnosis and specific therapy is of utmost importance in the everyday life of the patient.

In vitro study of rat prostate 5 alpha-reductase activity and its inhibition.

A simple and rapid method of measuring 5 alpha-reductase (5 alpha-R) activity and of determining the kinetic parameters (KM and Vmax) of the enzyme is described. The 5 alpha-R activity in the homogenate of the prostate of Wistar rats aged 8-12 weeks was established, and the effects of natural and synthetic steroids and of non-steroidal antiandrogens (IC50) upon the 5 alpha-R activity were studied. Of the natural steroids, 17-OH-progesterone was found to have the highest inhibitory effect (IC50 = 1.35 microM), followed in decreasing order by progesterone (IC50 = 5.0 microM) and 4-androstene-3,17-dione (IC50 = 21.6 microM). Oestradiol-17 beta had practically no inhibitory effect. Of the synthetic steroids, 4-MA had the highest inhibitory effect (IC50 = 0.068 microM), followed by nortestosterone (IC50 = 7.4 microM) and RU-486 (Mifepristone) (IC50 = 115 microM). Even at 1000 microM, cyproterone acetate exerted no inhibitory effect. Of the nonsteroidal compounds, ketoconazole proved a weak inhibitor (IC50 = 115 microM), while flutamide was practically ineffective.

Effect of treatment with Prolan or with a GnRH superactive analog on the sexual function of sows after weaning.

On a large, closed pig farm using artificial insemination (AI), 29 sows were treated with Prolan S-öl injection (Bayer, FRG) and 31 sows with a GnRH superactive analog (Ovurelin, D-Phe6-GnRH-EA, Reanal, Hungary) 48 h after weaning. The effect of treatment on the sows' sexual function was monitored by serum progesterone radioimmunoassay (RIA). The conception rate in the control group (36 sows) was 69.4%. In the groups treated with Prolan and Ovurelin it was 79.3 and 71%, respectively. The use of Prolan S-öl injection markedly reduced the number of acyclic sows and of those having an irregular oestrous cycle. Treatment with the GnRH analog inhibited the manifestation of weaning-induced heat; subsequently, however, it induced a regular cycle in 30 out of the 31 sows treated.

[Simultaneous occurrence of selective ACTH deficiency , achalasia, alacrimia and hyperlipoproteinemia]. / Szelektív ACTH érzéketlenség, achalasia és alacrimia együttes elöfordulása hyperlipoproteinaemiával.

An extremely rare clinical syndrome on a 7-year-old girl is presented. Besides isolated glucocorticoid insufficiency, achalasia and alacrimia disturbance of the lipid metabolism was also detected--this is a special feature of this case. The details of the endocrine workup is discussed, providing clues for the possible pathomechanism. The correct diagnosis and specific therapy is of utmost importance in the everyday life of the patient.

[Increased androgen-binding capacity and decreased free androgen index in male alcoholics with liver disease]. / Emelkedett androgenkötö kapacitás és csökkent szabad androgen index alkoholos májbeteg férfiakban.

Androgen binding capacity in the serum of 30 male patients under 51 years of age suffering from chronic alcoholic liver disease was investigated with tritiated 5 alpha-dihydrotestosterone. The result received by saturation analysis is regarded as an indicator of the SHBG (sex hormone binding globulin) content of the blood. Testosterone was measured simultaneously and the free androgen index (FAI) was calculated. The diagnosis of liver disease was based on case histories, physical symptoms and laboratory abnormalities, in some cases on liver biopsy too. Age-matched control group consisted of 18 healthy males. The results showed markedly elevated androgen binding capacities (SHBG contents), testosterone values in the normal range and significantly decreased FAI, indicating insufficient amounts of biologically active testosterone.

Direct radioimmunoassay of serum progesterone for monitoring the reproductive status of large pig herds.

The reproductive status of sows was monitored by serum progesterone radioimmunoassay (RIA) in a closed pig herd of 1100 sows, producing for the market. The method proved suitable for monitoring reproductive events (progesterone production during lactation, effect of weaning on the resumption of cycling, successful and unsuccessful inseminations, abortion, etc.) and for diagnosing ovarian disturbances (acyclia, irregular cycle, etc.). A total of 813 inseminations were followed up to determine the time of return to oestrus. The serum progesterone values of blood samples taken from the v. cava cranialis on day 22 after insemination indicated that pregnancy was diagnosed with a reliability of 75.5%, while non-pregnant sows failing to return to oestrus were identified with a reliability of 96.1%. The economic benefits to be derived from the use of this method in a pig herd are discussed.

High-dose ketoconazole treatment in advanced prostatic cancer.

For purposes of testosterone suppression, high doses of ketoconazole were administered to 18 patients with advanced cancer of the prostate for an average of 6 months. A well-evaluable response was observed both objectively and subjectively. The blood levels of the steroid hormones were measured during the treatment period and it was found that testosterone biosynthesis was blocked. Mild gastrointestinal symptoms occurred in the minority of the cases, and administration of the drug had to be stopped in one case because of jaundice. The possibility of the use of smaller doses is being investigated, which may lead to the development of an alternative to the endocrine treatment methods.