Possible modulating action of plant infusion of Ocimum sanctum L. on chromosomal aberrations and sister chromatid exchanges induced by chlormadinone acetate in human lymphocytes in vitro.

Chlormadinone acetate (CMA) is a synthetic progesterone analogue. It has its usage in oral contraceptives formulations and also for estrous synchronization of animals. The aim of the present study is to study the anti- genotoxic activity of the plant infusion against the CMA induced genotoxic damage on cultured human lymphocytes, using chromosomal aberrations and sister chromatid exchanges (SCFs) as parameters. For chromosomal aberration analysis, the treatment of 40 microM of CMA was associated with 4.33% abnormal metaphases. The treatment of 40 microM of CMA, separately with 1.075 x 10(-4), 2.125 x 10(-4) and 3.15 x 10(-4) gm l(-1) of plant infusion results in the reduction of the number of abnormal metaphases i.e. 2.67%, 2.00% and 1.67% respectively. For sister chromatid exchange analysis, the frequency of sister chromatid exchange per cell (SCE(S)/Cell) for the treatment of 40 microM of CMA was 6.43. The treatment of 40 microM of CMA, separately with 1.075 x 10(-4), 2.125 x 10(-4) and 3.15 x 10(-4) gm l(-1) of plant infusion results in the significant reduction of the frequency of SCE(S)/Cell i.e. 3.76, 3.01 and 2.94, respectively, as compared to the CMA (40 microM) treatment alone (6.43). The used dosages of plant infusion did not increase chromosomal aberrations and sister chromatid exchanges at significant level as compared to the untreated. The results of the present study suggest that the plant infusion per se does not have genotoxic potential, but can modulate the genotoxicity of chlormadinone acetate in human lymphocytes in vitro.

Effect of bufalin and pregnanes on vasoreactivity of human resistance arteries.

Endogenous Na/K ATPase inhibitory activity has been implicated in salt and water homeostasis in mammals and amphibians. Recent interest has focused on endogenous cardiac glycosides, some progesterone derivatives (pregnanes) and the amphibian bufodienolides. This study has examined the effects of non-planar and planar pregnanes and the bufodienolide bufalin on vasoreactivity of human resistance arteries. Bufalin and a non-planar pregnane caused concentration-dependent potentiation of the tone of submaximally pre-contracted arteries and inhibited endothelium-dependent relaxation, whereas a planar pregnane affected neither response. The relative potency of the compounds studied suggest the results do not simply reflect degrees of Na/K ATPase inhibition. The active compounds may be important in the regulation of vascular tone.

Some substances with proposed digitalis-like effects evaluated on platelet functions sensitive for cardiac glycosides.

1. The effects of progesterone, corticosterone 21-acetate, chlormadinone acetate, dehydroepiandrosterone 3-sulfate and lysophosphatidylcholine were tested on 86Rb-uptake, 3H-5-HT-uptake, ADP-induced aggregation and 5-HT-induced shape change in human platelets. Ouabain and digoxin were used for reference. 2. Ouabain and digoxin 10(-5) M inhibited 86Rb-uptake by more than 85%, and chlormadinone acetate 10(-5) M by 20%. The other substances had no effects. 3. Ouabain and digoxin were potent inhibitors on 3H-5-HT-uptake, whereas chlormadinone acetate had no effect. 4. Ouabain and digoxin increased ADP-induced aggregation but chlormadinone acetate decreased it. 5-HT-induced shape change was decreased by ouabain and digoxin, and to a lesser extent by chlormadinone acetate and its vehicle (ethanol 1.0%).

Interaction of progesterone derivatives with the digitalis target enzyme: impact of glycosidation on inhibitory potency.

As a function of the structural modification of the steroid nucleus, the inhibitory interaction of 11 progesterone derivatives with human Na/K-ATPase (Na+/K(+)-transporting ATPase, EC 3.6.1.37), through C3-O-rhamnosylation, is either much decreased or weakly up to strongly increased, so that the rhamnosyl residue contributes to the complementary Gibbs energy of interaction, at the most, the same Gibbs energy increment as realized in ouabain. After C3 beta-O-rhamnosylation, the activity of some progesterone derivatives considerably surpasses that of 3 beta-O-rhamnosyl-chlormadinolacetate, which has been known to elicit positive inotropy in cats. The progesterone derivatives (aglycons and glycosides), that have been analysed more closely, produce their effects by the same molecular mechanism of interaction with Na/K-ATPase as characteristic for digitalis aglycons and glycosides. The results promise to pave the way for the identification of the chemical nature of endogenous digitalis and for the design of novel inotropic drugs.

[Morphologic and biochemical changes in the hypothalamus in hormonally stimulated sheep]. / Morfologické a biochemické zmeny v hypotalame u hormonálne stimulovaných oviec.

The effect of exogenous hormonal preparations (PMSG, SG, PMSG + PGF2 alpha and FSH) was studied as exerted on neurosecretion and on changes in the nuclear volume of nucleus paraventricularis, nucleus ventromedialis, nucleus infundibularis and nucleus tuberomamillaris, and on histological changes in the third cerebral ventricle. Changes in catecholamine concentrations were determined in the rostral, medial, and caudal hypothalamus of the ewes. The study was performed with 28 ewes of the Slovak Merino breed during the oestrus period. The oestrus was synchronized by the Agelín vaginal tampons (Spofa). The ewes of four experimental groups were stimulated by the administration of 1000 I.U. PMSG (Antex-Leo), 1000 I. U. Serum Gonadotropin (Bioveta Ivanovice na Hané), 1000 I. U. PMSG with 250 micrograms Oestrophan inj. (Léciva Praha), and 450 I. U. Folistiman (VEB Arzneimittelwerk Dresden, GDR). The brain samples were processed by common histological methods. A caryometric analysis was performed at 3000-fold magnification and by measurement of 200 cells of one sample. The data were subjected to mathematical processing after Fischer and Inke (1956). The amount of neurosecretory material was evaluated by light microscopy (Nakahara, 1963). The multiplication of neurosecretion in the hypothalamic nuclei testifies to the fact that the administration of hormonal preparations stimulates the function of hypothalamic structures, which corresponds with the results of the caryometric analysis. It also ensues from the results that the administration of serum gonadotropins (PMSG and SG) at the dose of 1000 I. U. results in a statistically significant decline (P less than 0.001) of the concentrations of norepinephrine in the rostral, medial and caudal hypothalami of the ewes, although the PMSG preparation has a more pronounced effect when compared with SG. The hormonal stimulation with PGF2 alpha and with FSH causes no significant changes in the concentration of norepinephrine in the hypothalamus of the ewes. As norepinephrine is considered as a neurosecretion inhibitor, the multiplication of neurosecretion in the hypothalamus nuclei after stimulation with serum gonadotropins correlates with the decline of hypothalamic norepinephrine concentrations.

Antiandrogenic effects of cyproterone acetate and chloromadinone acetate on the rat hypothalamus as revealed by electroencephalographic responses.

Bipolar stainless steel electrodes were implanted stereotaxically into given hypothalamic and cortical regions of the brain of 48 rats. The animals were divided into two groups of equal numbers and the electrical activity of the given regions was recorded electroencephalographically (EEG). The effect of the acute and chronic administration of cyproterone acetate (CPA) or chloromadinone acetate (CHA), in doses inducing sterility, on EEG activity were observed. Apart from slight inhibition in the preoptic region, no appreciable EEG changes were found after CPA, while CHA reduced the discharge frequency in both the preoptic and the posterior hypothalamus. These findings can be attributed to the presence of two different types of androgen receptors in the hypothalamus.

LH and estradiol concentration in the blood and morphological characteristics of the hypothalamus, hypophysis and the ovaries of sheep after LH/FSH-RH and synthetic gestagen treatment.

In the experiments the effect of synthetic LH/FSH-RH on the concentration of LH and estradiol 17-beta (E2) in the blood of sheep was followed during physiological anestrus, and the preovulatory LH release and the E2 level in the blood serum after the synchronization of estrus with chlormadinone acetate (CAP) were studied. Furthermore, the nuclear volumes of the neurons in the ncl. hypothalamicus anterior (NHA), ncl. paraventricularis (NPV), ncl. arcuatus (NARC) and the pars distalis ncl. mamillaris medialis (PD NMM) as well as the percentage numbers of the adenohypophyseal cells and the number of normal and atretic tertiary follicles of the same ewes are described. The results show that after synthetic LH/FSH-RH treatment the LH concentration gradually increases in the blood, intravaginally instilled CAP affects the hypothalamic neurons probably by increasing significantly the concentration of the endogenous E2 that sensitizes the receptors of the hypothalamo-hypophyseal structures to LH/FSH-RH, enhancing, thus, its effect. The changes in the nuclear volume of the neurosecretorial cells of the hypothalamus after the administration of the mentioned substances indicate an ultrashort feed back in the regulation of secretion and the LH/FSH-RH release by the NARC cells. The changes were considered from the viewpoint of influencing the estrous cycle and the productivity of ewes.

[Quantifiable morphokinesis on parts of the hypothalamo-adenohypophyseal-gonadal axis of hormonally castrated boars. I. Studies on boars treated with chlormadinone acetate]. / Zur quantifizierbaren Morphokinese an Gliedern der hypothalamo-adenohypophyseo-gonadalen Achse hormonell desexualisierter Eber 1. Mitteilung: Untersuchungen an mit Chlormadinonazetat behandelten Ebern.

A clearly morphokinetic effect was recorded from the parts of the hypothalamo-adenohypophyseo-gonadal axis in boars that had been treated with chloromadinone acetate (CAP). Luteinising hormone release is inhibited by CAP, which is reflected in an increase of luteinising hormone cells in their storage form. This leads to statistically significant decline in androgenic testicular tissue percentage and in the cell nucleus volumes of Leydig's cells. Excessive weight decrease of the testicle is caused primarily by some 60 per cent reduction in tube length. The neurons of the tested hypothalamic nuclear region, that is the nucleus praeopticus medialis, have shown that in treated animals cell nucleus volumes were significantly depressed.

Sex hormone-dependent brain maturation and sexual behaviour in rats.

In male and female rats the endogenous steroid and gonadotrophin secretion was inhibited by injecting high doses of chlormadinone acetate (CmAc) from day 14 to 24 of life, i. e. during the period of brain maturation. In adulthood the males treated prepubertally with CmAc exhibited reduced sexual activity and fertility, whereas the females did not differ from the controls. More complete sex hormone deficiency during brain maturation was achieved by castration on day 14 of life. Controls were castrated at normal puberty time (40--60 days). Both groups were then substituted with androgens or oestrogens. In the females castrated on day 14 no impairment of sexual behaviour was observed as compared to the later castrated controls. In contrast, the early castrated males showed delayed onset of mounting behaviour. At autopsy, the weights of their sex organs were found to be lower than in the controls despite equal testosterone replacement for several months. These findings speak in favour of a permanently diminished responsiveness to androgens in males having been exposed to more or less severe androgen deficiency during sex specific brain maturation. Hence, the maturation of a male hypothalamus as well as the differentiation appears to depend at least in part on the presence of androgens, whereas in females it runs without hormonal influence.

PIP: Sex hormone-dependent brain maturation and sexual behavior in rats was studied in animals castrated at an early age and in animals treated with an antigonadotropic and antiandrogenic drug. 30 male and 25 female rats were treated with 6 mg chlormadinone acetate (CMA) from Days 14 to 24 of life. Male and female sexual behavior was tested. In adulthood these treated males were stimulated with 5 IU of follicle stimulating hormone plus 30 IU human chorionic gonadotropin daily and sexual behavior observed. Males treated with CMA exhibited reduced sexual activity and fertility, whereas females were similar in response to controls. In a 2nd experiment, males and females were castrated on Day 14 of life. Other animals were castrated at the start of sexual activity. Both groups then received androgens or estrogens. Sexual behavior was again tested. In females castrated by Day 14, sexual behavior was unimpaired as compared with the older castrates. However, early castrated males showed delayed onset of mounting and ejaculatory behavior after beginning androgen replacement. The reduced mounting frequency and ejaculatory behavior were permanently observed during 2-4 months of hormone replacement. At autopsy the weights of the sex organs were lower than controls despite equal testosterone regimen. In males exposed to severe androgen deficiency during sex-specific brain maturation there is a permanently diminished responsiveness. Female maturation occurs without hormonal influence.

A four-year evaluation of the chronic toxicity of megestrol acetate in dogs.

PIP: A 4-year evaluation of the chronic toxicity of megestrol acetate in dogs is reported. .01, .1 or .25 mg of megestrol acetate/kg/day or .25 mg of chlormadinone acetate/kg/day was administered orally for 4 years t o female beagle dogs. The hormone-treated dogs tended to gain more weig ht than did the controls (controls vs. .25 mg megestrol acetate every month after the 3rd p less than .01). All treated dogs revealed decreased evidence of estrus. Mucoid vaginal discharges were more prevalent among the middle and high dose groups. Mean hemoglobin, packed cell volume and total erythrocyte values were slightly decreased while mean total leucocyte count and erythrocyte sedimentation rates were slightly increased in the middle and high dose groups. Clotting me chanism did not reveal any disturbances. Evidence of diabetes consistin g of bilateral cataracts, elevated serum glucose concentrations and glycosuria after 4 years in 2 of 16 high-dose megestrol acetate and in 6 of 15 chlormadinone acetate-treated dogs was revealed. It is concluded that the effects of megestrol acetate were similar but less severe than those of chlormadinone acetate.^ieng

The action of chlormadinone acetate on the estradiol uptake by rat organ cultures.

PIP: The action of chlormadinone acetate on the estradiol uptake by rat organ cultures was investigated. Rat hypothalami and uteri tissue were incubated in TC 199 medium for 49 or 72 hours with tritiated estradiol-17beta and unlabeled estradiol or chlormadinone acetate. Chlormadinone acetate added simultaneously to tritiated estradiol inhibited the estradiol uptake (p less than .01) and when added 24 hours previously, the uptake increased (p less than .01) in both uterus and hypothalamus. Unlabeled estradiol also diminished (p less than .01) the estradiol uptake when added simultaneously and increased its uptake when added prior to tritiated estradiol-17beta. These results may indicate an estrogeniclike effect of chlormadinone acetate, perhaps by increasing the binding sites.^ieng