Does the bone morphogenetic protein 7 inhibit oocyte maturation by autocrine/paracrine in mud crab?

A bone morphogenetic protein ligand (BMP7) and its two receptors (BMPRIB and BMPRII) were recently cloned and characterized in the mud crab, Scylla paramamosain. However specific functions of BMP7 and the mechanistic pathways regulating its function are largely unidentified. In the present study, we separated oocytes and follicle cells from the ovarian explants of S. paramamosain. Subsequent analysis using semi-quantitative PCR demonstrated that the mRNA of Sp-BMP7 was exclusively expressed in follicle cells while Sp-BMPRs were expressed in both oocytes and follicle cells. In vitro experiments further showed that the mRNA and protein levels of Cyclin B increased but Sp-BMP7 declined in 17α, 20ß-Dihydroxyprogesterone (DHP)-induced oocytes. Furthermore, the inhibitory effects of Sp-BMP7 were not affected by the elimination of the contact/gap junction-mediated communication between oocytes and follicle cells. Our data indicate that BMP7 may play a role in the suppression of DHP-induced oocyte maturation by affecting autocrine/paracrine pathways in S. paramamosain.

Multicentred clinical study of the metabolic effect of the monthly injectable contraceptive containing dihydroxyprogesterone acetophenide 150 mg + estradiol enanthate 10 mg.

The metabolic effect of the monthly injectable contraceptive containing dihydroxyprogesterone acetophenide (DHPA) 150 mg + estradiol enanthate (EEn) 10 mg was compared to that of other regularly used contraceptive methods (pills containing: ethinylestradiol (EE) 0.050 mg + levonorgestrel (LNG) 0.250 mg, EE 0.030 mg + LNG 0.150 mg; EE 0.030/0.040/0.030 mg + LNG 0.050/0.075/0.0125 mg; norethisterone enanthate (NEE) 200 mg i.m.; non-hormonal methods). Serum triglycerides, HDL/LDL-cholesterol, copper, ceruloplasmin, total and free cortisol, CBG, total and free testosterone and SHBG in chronic users were determined. A total of 237 women took part in this study. Taking users of non-hormonal methods as control, triglyceride levels were higher, and total and free testosterone levels were lower in women using DHPA 150 mg + EEn 10 mg and in those taking contraceptive pills (p less than 0.05 - 0.01). Such modifications were slightly less in the group using the injectable. The effects of DHPA 150 mg + EEn 10 mg on HDL/LDL-cholesterol copper, ceruloplasmin, CBG, total and free cortisol and SHBG were rare or non-existent. Nevertheless, the contraceptive pills (even the low-dose formulations) correlate with modifications of all those variables, which were highly significant in comparison with the injectable (p less than 0.01) and with non-hormonal methods (p less than 0.01); there were no differences between the last two methods. The results suggest that the metabolic effect of DHPA 150 mg + EEn 10 mg is not higher than that of the commonly used oral contraceptives. On the other hand, they do not suggest that the dose contained in this injectable is exaggerated. There is no evidence that it produces accumulation of effects in the organism. These findings should be taken into account when referring to the long-term safety of this injectable.

Development of sensitivity to maturation-inducing steroids and gonadotropins in the oocytes of the tobinumeri-dragonet, Repomucenus beniteguri, Callionymidae (Teleostei).

The effectiveness of steroids and gonadotropins (GtHs) in inducing final oocyte maturation was examined at different times of the day (0100, 0500, 0700, 0900, 1100, 1300, 1700, 2100 hr) in a daily spawning marine teleost, the tobinumeri-dragonet, Repomucenus beniteguri. The responsiveness of oocytes to GtHs and steroids was different at various times of day. The sensitivity of oocytes to hormones was apparent only during a certain period of the day (0100-0900 hr). Germinal vesicle breakdown (GVBD) could be induced by GtHs but not by steroids at 0100 hr. At 0500 hr, not only GtHs but also steroids at higher doses induced GVBD and ovulation. Oocytes underwent GVBD and ovulation at low doses of steroids at 0900 hr. GVBD and ovulation spontaneously occurred at 1100 and 1300 hr without any hormonal treatment. In addition, diameter and histological changes of ovarian oocytes were investigated. Ovarian oocytes were smaller than 420 microns, and only one oocyte group existed in distribution of oocyte frequency at 2100 and 0100 hr. With the increase in size of oocytes, two distinct groups appeared at 0500 hr. Larger oocytes completed GVBD at 1100 hr. Ovulation occurred between 1300 and 1700 hr and oviposition was completed between 1700 and 2100 hr daily. These results clearly show that the oocyte of the dragonet possesses a daily maturation rhythm. Responsiveness of oocytes to GtHs appeared earlier than responsiveness to steroids. This suggests that sensitivity to steroids is induced by GtH.

In vitro luteinizing hormone-releasing hormone release from superfused rat hypothalami: site of action of progesterone and effect of estrogen priming.

The study examined the effect of estrogen priming on progesterone (P4)-induced LHRH release, the tissue site of action of P4, and the effect of 5 alpha-dihydroxyprogesterone (5 alpha-DHP) on LHRH release from hypothalamic fragments superfused in vitro. Immature female rats were ovariectomized (OVX) and, at 28 days of age, Silastic capsules containing estradiol (E2) were implanted. Two days later, animals were killed and hypothalamic fragments were removed and transferred to superfusion chambers. The hypothalamic units received P4 or 5 alpha-DHP delivered in an intermittent mode (10-min on, 20-min off). LHRH was determined in perfusates by RIA. After the input characteristics of different infusion modes (single pulses, intermittent, and continuous) of P4 infused into superfusion chambers were assessed, an intermittent infusion mode (10-min on, 20-min off) was selected for further examinations. In the mediobasal hypothalamic-anterior hypothalamic-preoptic area (MBH-AHA-POA) tissue preparations, we observed: 1) an infusion of 5 alpha-DHP was ineffective in stimulating LHRH release; 2) the release pattern of LHRH in response to three different P4 doses (10, 20, and 50 ng/ml) was similar in terms of percent changes (202% to 219% over control values); and 3) E2 priming was absolutely required for P4-stimulated LHRH release, and this requirement appeared to be dose dependent. Upon an examination of three hypothalamic tissue boundaries [the MBH, the POA-suprachiasmatic nuclei (POA-SCN), and the median eminence (ME)] to better delineate the in vitro site of action of P4 on LHRH release, it was demonstrated that the MBH responded upon P4 infusion, whereas the POA-SCN was unable to do so. The ME also responded upon P4 infusion, and LHRH release followed closely the pulsatile administration of P4 since upon each challenge of the steroid at the concentration of 10 or 20 ng/ml, a significant rise in LHRH release occurred. However, the temporal patterns of LHRH release from the ME appears to be different from those obtained from the MBH as well as the MBH-AHA-POA. These observations demonstrate that an intermittent infusion of P4, but not 5 alpha-DHP, is effective in activating the neural LHRH apparatus. Estrogen is an obligatory requirement for this P4-stimulated LHRH release, and the neural site of action of P4 resides within the MBH. However, this steroid also can act directly upon the ME nerve terminals to release LHRH.

Tixocortol pivalate, a corticosteroid with no systemic glucocorticoid effect after oral, intrarectal, and intranasal application.

Tixocortol pivalate is a corticosteroid with topical anti-inflammatory activity equal to that of hydrocortisone. It was evaluated in a group of 18 normal subjects to determine whether it exerted any systemic glucocorticoid activity after single oral or intrarectal doses and after short-term dosing by the intranasal route. Effects of tixocortol pivalate were compared to those of oral dexamethasone and intrarectal betamethasone 21-phosphate. By the three routes, tixocortol pivalate does not induce any changes in plasma cortisol, leukocyte counts (neutrophils, lymphocytes, monocytes, eosinophils), blood glucose, or 24-hr urinary excretion of sodium and potassium, whereas there were changes after dexamethasone and betamethasone. Tixocortol pivalate, however, increased urinary free cortisol-like substances. It is concluded that tixocortol pivalate given for short periods by nonparenteral routes does not induce a measurable systemic glucocorticoid effect.

Pharmacological study of a new anti-inflammatory steroid, tixocortol pivalate (JO 1016).

Some pharmacological activities of pregn-4-ene-3,20-dione-21-thiol-11 beta,17 alpha-dihydroxy-21-pivalate (tixocortol pivalate, JO 1016 Pivalone), a new steroidal anti-inflammatory compound, are described. The anti-inflammatory activity of tixocortol pivalate has been clearly demonstrated in various tests using adrenalectomised and intact animals. Of particular interest is the dissociation of its local and systemic activities. Comparison with hydrocortisone acetate indicates a similar or greater anti-inflammatory activity, by either the local or topical routes, but tixocortol pivalate is between 60 and 300 times less active than hydrocortisone acetate after oral or subcutaneous administration. Glucocorticoid activity was only detected at very high oral doses of tixocortol pivalate and the highest tested subcutaneous dose (300 mg/kg) failed to induce significant activity. Hydrocortisone acetate exerted glucocorticoid effects at much lower doses. It is possible therefore the local or topical use of tixocortol pivalate in therapy may not cause the unwanted side effects of many of the corticosteroids in current use.

A comparison of the effects of tixocortol pivalate (JO 1016), beclomethasone dipropionate and hydrocortisone acetate on the activation of lymphocytes.

A comparative study was made of the effects of hydrocortisone acetate, beclomethasone dipropionate and a corticosteroid substitute (pregn-4-ene-3,20-dione-21-thiol-11 beta,17 alpha-dihydroxy-21-pivalate, tixocortol pivalate, JO 1016, Pivalone) on lymphocyte activation. The response measured in vivo was the production of heamagglutinating antibody by mice in response to immunization with sheep erythrocytes. The in vitro response tested was the blastocytic transformation of T cells in mixed lymphocyte culture. All three compounds suppressed T cell activation in vitro. Tixocortol pivalate however, was non-toxic and non-immunosuppressive when administered to mice, in contrast to hydrocortisone acetate and beclomethasone dipropionate, which were toxic and immuno-suppressive at the highest dose levels tested. These results indicated that tixocortol pivalate would, by virtue of its low toxicity and absence of general immunosuppression, have potential advantages over both hydrocortisone acetate and beclomethasone dipropionate in the topical treatment of allergic conditions of mucous membranes.

A comparison of the effects of tixocortol pivalate (JO 1016), hydrocortisone acetate and beclomethasone dipropionate on the phagocytosis and lysis of microorganisms by alveolar macrophages.

A comparison was made of the ability of guinea pig alveolar macrophages, which had been pretreated with hydrocortisone acetate, beclomethasone dipropionate or a corticosteroid substitute pregn-4-ene-3,20-dione-21-thiol-11 beta,17 alpha-dihydroxy-21-pivalate (tixocortol pivalate, JO 1016, Pivalone), to phagocytose and lyse Staphylococcus aureus or Candida albicans. The three drugs had different patterns of effect on phagocytosis and lysis. Hydrocortisone acetate had little effect on the phagocytosis of Staph. aureus at any dose level tested, but the two higher concentrations slightly inhibited intracellular lysis; phagocytosis of C. albicans was inhibited initially but increased at 5 h. Both beclomethasone dipropionate and tixocortol pivalate caused an initial stimulation of phagocytosis and lysis of Staph. aureus. Ingestion of C. albicans was increased in macrophages pretreated with beclomethasone dipropionate, but their fungicidal activity was unchanged. Pretreatment with tixocortol pivalate stimulated the initial phagocytosis of C. albicans but continued uptake on prolonged incubation was inhibited. Lysis of the ingested organisms was not markedly affected. Since the production of any effect on the phagocytic and lytic activity of alveolar macrophages required much higher levels of tixocortol pivalate than of either of the other two drugs, it is suggested that in clinical use correspondingly higher doses of tixocortol pivalate could be given without danger of affecting either phagocytic activity or the immune response.

A comparison of the effects of tixocortol pivalate (JO 1016), hydrocortisone acetate and beclomethasone dipropionate on collagen synthesis and degradation.

The effects of a corticosteroid substitute pregn-4-ene-3,20-dione-11 beta,17 alpha-dihydroxy-21-pivalate (tixocortol, JO 1016, Pivalone), hydrocortisone acetate and beclomethasone dipropionate on collagen metabolism in mouse calvariae were compared by histological and biochemical determination of collagen content, and by radio-assays of collagenase and collagenase inhibitor. Hydroxyproline assay revealed dose-related increases in collagen content/wet wt. tissue. These were greatest in explants treated with beclomethasone dipropionate, tixocortol pivalate being effective only at much higher concentrations than the other 2 drugs. Hydrocortisone acetate and beclomethasone dipropionate stimulated collagenase production at the lowest levels tested whereas tixocortol pivalate was only slightly stimulatory at the highest treatment level. These results suggest that in clinical use tixocortol pivalate would be much less likely than hydrocortisone acetate or beclomethasone dipropionate to cause degenerative changes in connective tissue.

Inhibitory effects of various progestins and deoxycorticosterone on the rapid onset of maternal behavior induced by ovariectomy-hysterectomy during late pregnancy in rats.

The inhibitory effects of progesterone (P), 5 alpha-dihydroprogesterone (5 alpha-DHP), 17 alpha-hydroxyprogesterone (17-HP), and deoxycorticosterone (DOC) upon the rapid onset of maternal behavior induced during late pregnancy in primigravid rats by ovariectomy-hysterectomy (OH) were examined. Progesterone administration at a dosage of 5.0 mg oil vehicle daily (beginning on Day 17 and ending when the subject responded maternally to foster pups) significantly delayed by about 1.5 to 2.0 days the onset of maternal behavior. In contrast, P at dosages of either 1.0 or 2.5 mg daily, 5 alpha-DHP at 5.0 mg administered daily in either an oil or Tween-80 vehicle, 17-HP at 5.0 mg in oil daily, and DOC at dosages of either 5.0 or 10.0 mg in oil daily failed to effect the rapid onset of maternal behavior induced by ovariectomy-hysterectomy on Day 17 of pregnancy. These data suggest that during pregnancy the onset of maternal behavior is inhibited in a rather specific manner by progesterone.

Oestrogen formation from C19 precursors in human choriocarcinoma in culture.

A cloned cell line of human choriocarcinoma was evaluated as a model of human placental oestrogen production. Oestrone formation from dehydroepiandrosterone (D), D-sulphate (DS) or 4-androstenedione (A) was less than or equal to 5% of oestradiol-17beta (Oe2) formation. Oe2 formation from D and A was similar (100-150 pmole/h/10(7) cells); that from DS was 10 times less. Omitting serum from the medium increased Oe2 yield from DS 4-fold; addition of albumin restored these yields to control values (P greater than 0.05, t-test), presumably by binding DS. N6,O2'-dibutyryl-adenosine 3',5'-cyclic monophosphoric acid and theophylline treatment for 72 h stimulated (P less than 0.01) Oe2 formation from D (36%), DS (66%) and A (183%). In intact cells, sulphatase activity, Oe2 formation from D and Oe2 formation from DS equalled those in homogenates (P greater than 0.05) but Oe2 formation from D was greater than that from DS in both systems (P less than 0.001), indicating a deficiency of sulphatase relative to subsequent enzymes of oestrogen synthesis. Steroids, at concentrations previously shown to inhibit placental sulphatase or 3beta-hydroxysteroid dehydrogenase, did not inhibit choriocarcinoma enzymes. Except for its relative sulphatase deficiency and insusceptibility of oestrogen synthesizing enzymes to steroid inhibitors, choriocarcinoma appears to be a useful model of placental oestrogen synthesis.

Further unravelling of the causes of ACTH-induced hypertension in the sheep.

1. Acute severe sodium subtraction (20-25% of total exchangeable sodium) before or during treatment with adrenocorticotrophic hormone (ACTH) does little to modify the increase in blood pressure induced by ACTH. 2. Chronic low salt diet, less than 5 mmol/day, abolishes the blood pressure increase, but the response can be restored by increasing the sodium intake to as little as 10 mmol/day. 3. 17alpha,20alpha-Dihydroxyprogesterone infused concurrently with other adrenal steroids will mimic ACTH hypertension and perhaps represents a new class of steroid capable of influencing blood pressure.

Effect of some progestational steroids on lactation in Egyptian women. I. Milk yield during the first year of lactation.

PIP: The effect of monthly injectios of 300 mg Depo-Provera or 150 mg Deladroxone, and of daily oral administration of .5 mg chlormadinone acetate or .3 mg quingestanol acetate on lactation was studied in Egyptian women during the 1st year of lactation. Women receiving Depo-Provera had the highest milk yield, followed by those taking Deladroxone and quingestanol acetate. The milk yields while taking these hormonal preparations were higher than in untreated controls. However, those women taking chlormadinone acetate had lower milk yields than untreated controls. The increased milk yield is probably due to the progestagenic activity, and minimal estrogenicity, of these drugs.^ieng

[Response of peripheral receptors to the parenteral administration of an association of dihydroxyprogesterone acetophenide and estradiol-3 benzoate-17-n-butyrate. Preliminary note]. / La risposta dei recettori periferici alla somministrazione parenterale dell'associazione diidrossiprogesterone acetofenide estradiolo-3-benzoato-17n-butirrato. Nota preventiva

PIP: The basal temperature curves, cervical mucus crystallation, endocrin e colpocytology, and endometrial cytology (Endo-Cyte) in 11 women over 30 cycles was determined following the im administration on the 8th day of the cycle of an estroprogestinic combination of 150 mg of dihydroxyprogesterone acetophenide and 10 mg of estradiol-3-benzoate-17-n-butyrate. The findings confirmed the antiovul atory action of the preparation, the absence of unwanted side effects, s ecretory transformations in the endometrium, the normality of the menstrual cycle, and the advantages associated with a single monthly administration.^ieng