Effects of the antifertility drug enovid in five strains of mice, with particular regard to carcinogenesis.

PIP: The antifertility drug, Enovid, was tested for possible carcinogenicity in female mice of 5 specially selected strains: BALB/c, C3H, C3HfB, A, and C57BI. Enovid was chosen for testing since it is one of the most widely used oral contraceptives. The 5 strains of mice provided maximum genetic variation in the test animals. The drug was fed at 3 dose levels: 5 mcg/gm, 10 mcg/gm and 20 mcg/gm of food. The lowest dose did not prevent reproduction. The 10 mcg dose prevented some females from reproducing. The 20 mcg dose prevented all females from reproducing. The strains of mice differed in their response to Enovid. Weight gain was reduced in all strains. Effect on life-span varied, partly because of the tumors. Cervical and vaginal lesions showed invasion of the epithelium into the stroma but was limited, with few exceptions to the BALB/c females. In the BALB/c strain these lesions occurred in controls as well, but showed more progression and a higher incidence with the highest dose of Enovid. None of these lesions appeared grossly as tumors and none had extended beyond the vaginal wall or metastasized. They were observed only on histologic sections. Neither ovarian nor mammary gland tumors were increased in any strain. In the C3H strain such tumors seemed to be inhibited. In the C3HfB strain there was some inhibition of hepatomas and in the BALB/c strain some inhibition of adrenocortical adenoma. Chromophobe adenomas of the hypophysis were significantly increased in old C57BI females treated with the highest dose of Enovid. Offspring of Enovid-treated females showed no abnormalities. Enovid increased the occurrence and may have advanced the progression of epithelial lesions of the cervix and vagina of old BALB/c females. A study of the lesions in untreated females of this strain might help the understanding of carcinoma in situ in women and possibly the appearance of adeno-carcinoma of the vagina of young women whose mothers had been treated with stilbestrol during the first trimester to maintain pregnancy. Other neoplasms in this strain were not increased by the Enovid therapy. In the C3H strain mammary tumors were reduced by the Enovid and those that did occur were found later than in controls. Results from experimental animals should be applied to humans with care. Such results are of greatest value in directing attention to certain areas for investigation.^ieng

Estrogens and the hypothalamo-pituitary-adrenal axis in man: evidence for normal feedback regulation by corticosteroids.

Estrogen treatment and pregnancy are associated with higher than normal plasma free (nonprotein-bound) cortisol levels. In spite of this, clinical manifestations of steroid excess are not seen in these conditions. To explain this seeming discrepancy, it has been postulated that estrogens may induce tissue resistance to the actions of cortisol, and that one aspect of this resistance may be a higher set-point for ACTH suppression by corticosteroids. This possibility was studied in seven normal women. Plasma total and free cortisol levels as well as urinary cortisol excretion were measured during a control period and during treatment with ethinyl estradiol (100 micrograms/day). During both periods, graded doses of dexamethasone (0.2-mg increments; 0-1.4 mg/day) were administered. Estrogen treatment resulted in elevated plasma total and free cortisol levels, but urinary cortisol excretion was not affected. Dexamethasone administration resulted in a dose-dependent reduction of plasma total and free cortisol as well as urinary cortisol. The dose-response curve for suppression by dexamethasone of urinary cortisol during estrogen treatment was indistinguishable from that during the control period. The dose-response curve for plasma free cortisol suppression suggested that during estrogen treatment, slightly more dexamethasone was required to suppress free cortisol. However, this effect was small. In view of the overall data, we conclude that 1) estrogen does not increase integrated free cortisol prevailing in vivo; 2) estrogen does not significantly alter the hypothalamic or pituitary set-point for ACTH suppression by corticosteroid; 3) the elevation of plasma free cortisol is relatively minor and possibly an in vitro phenomenon; and 4) the present findings are compatible with the absence of clinical hypercorticism in hyperestrogenized states.

Biphasic change in pituitary capacity induced by estrogen in hypogonadal women.

PIP: 5 healthy postmenopausal women aged 52-59 years with elevated gonadotropin levels and low estradiol (E2) (10+ or -1.5 pg/mol) concentrations volunteered for this study which measured changes in responses to pulses of luteinizing hormone-releasing factor (LHF) (10 mcg at 2-hour intervals 3 times) and thyroid-releasing factor (TRF) (200 mcg at 2-hour intervals 3 times) for luteinizing hormone (LH), follicle stimulating hormone (FSH), and prolactin (PL) before and during administration of a large dose of ethinyl estradiol (EE) (400 mcg/day) for 5 days. Responses to LRF and TRF stimulation on the 2nd and 5th days of EE treatment were studied in 2 experimental periods in the same individual at an interval of at least 10 weeks between treatments. During 5 days of EE treatment, LH 1st declined (50%) and then returned to the original level, forming a previously observed U-shaped curve. The initial fall in basal FSH level was not followed by a return to pretreatment level. These changes were accompanied by an exponential increase of more than 3-fold in the PL levels. The bidirectional pattern of LH release was associated with parallel changes in pituitary sensitivity to a 10-mcg pulse of LRF. Pituitary reserve, defined as the response to the 2nd and 3rd pulses of LRF, exhibited unidirectional augmentation. This progression in PL release upwards was unaccompanied by changes in pituitary PL sensitivity and reserve. The contribution of hypothalamic LRF and dopamine in the participation of these functional changes of LH, FSH, and PL are discussed.^ieng

Chorea induced by oral contraceptives.

A rare complication of oral contraceptive therapy is the induction of chorea. We here describe five cases of chorea in patients receiving low- or high-dose estrogen-containing contraceptives. All patients were nulliparous, young (average age 19 years), and became symptomatic shortly (average of 5 weeks) after initiation of contraceptive therapy. Two patients previously suffered an episode of Syndenham chorea; one experienced chorea in the course of Henoch-Schönlein purpura; and two had a history of congenital cyanotic heart disease without chorea. Dyskinesia resolved in all patients upon discontinuing the medication. Patients with preexisting striatal abnormalities appear more susceptible to oral contraceptive-induced chorea which is reversible on drug discontinuation. The mechanism of oral contraceptive-induced chorea is unknown, but clinical and experimental data suggest that it involves altered central dopaminergic activity.

PIP: It is known that chorea is a rarely-occurring complication of oral contrceptive therapy. 5 case histories of chorea in patients receiving either low- or high-dose estrogen-containing contraceptives are reviewed. All the patients were young and nulliparous. They developed the symptoms within an average of 5 weeks after therapy initiation. Dyskinesia ceased upon cessation of the oral contraceptive therapy. A summary of 17 previously-reported cases of oral contraceptive-associated chorea is also presented in tabular form. These cases plus the 5 reviewed in this paper suggest that chorea arises in women with abnormalities of the basal ganglia of various etiologies and will probably not occur in normal individuals. Studies with animals have indicated that female sex hormones may enhance central dopaminergic sensitivity, bringing on chorea in oral contraceptive patients. In the 3 of 5 patients here described who had previously experienced an episode of chorea, the contraceptive-induced disorders, i.e., asymmetries, orofacial involvement, and personality changes, were similar to the original movement disorders. The other 2 cases studied here had not experienced a chorea episode but did have a history of neonatal cyanosis.

Antiepileptic medication and oral contraceptive interactions: a national survey of neurologists and obstetricians.

Hepatic enzyme-inducing antiepileptic drugs (AEDs) lower oral contraceptive (OC) sex hormone levels approximately 40% and increase the risk of unplanned pregnancies in women with epilepsy. AEDs also increase the risk of birth defects in offspring of women with epilepsy. We performed a national survey to determine obstetricians' and neurologists' knowledge of OC and AED interactions and the risk of birth defects for women with epilepsy taking AEDs. We received responses to a mailed questionnaire from 160 of 1,000 neurologists (16%) and 147 of 1,000 obstetricians (15%) from 47 states. Practice demographics and ages of responders were typical for U.S. neurologists and obstetricians. Ninety-one percent of neurologists and 75% of obstetricians said they treat women with epilepsy of child-bearing age. Only 4% of the neurologists and none of the obstetricians, however, knew the effects of the six most common AEDs on OCs, even though 27% of neurologists and 21% of obstetricians reported OC failures in their patients taking AEDs. Although increasing OC doses can compensate for insufficient OC sex hormone levels due to AEDs, most physicians do not increase the doses. Even though the risk of birth defects for the offspring of women with epilepsy is 4 to 6%, up from the background level of 2%, 44% of neurologists thought the risk was lower (0 to 3%), and some of the respondents guessed that it was as high as 50%. Many neurologists and obstetricians do not have accurate information to counsel women with epilepsy properly about their contraceptive and pregnancy choices.

PIP: Responses from 160 of 1000 neurologists (16%) and 147 of 1000 obstetricians (15%), selected from an American Medical Association listing to receive a mailed questionnaire, revealed a disturbing lack of knowledge about the interactions between antiepileptic medications and oral contraceptives (OCs). Hepatic enzyme-inducing antiepileptic drugs lower OC estradiol levels by about 40% and may reduce free progestin levels, thereby increasing the risk of unplanned pregnancy; moreover, antiepileptics increase the risk of birth defects in their epileptic users, who already have a 4-6% increased risk of such defects. Physicians can reduce, but not prevent, the risk of unwanted pregnancy by increasing the OC estradiol dose to at least 50 mcg and prescribing valproic acid and gabapentin (non-enzyme-inducing antiepileptics). 91% of neurologists and 75% of obstetricians reported that they treated epileptic women of childbearing age, and 27% of the former and 21% of the latter physicians acknowledged cases of OC failure in these patients. Only 4% of the neurologists and none of the obstetricians knew the effects of the 6 most common antiepileptic drugs on OCs. Just 41% of neurologists and 43% of obstetricians routinely had patients adjust their OC doses if they were taking antiepileptics. Such adjustment was more likely among physicians who had an epileptic patient with an unintended pregnancy and those who had accurate knowledge of OC-antiepileptic drug interactions. 44% of neurologists and 23% of obstetricians underestimated the birth defects risk as 0-3%. Since the physicians who chose to respond to this survey were presumably more concerned and knowledgeable about the reproductive effects of antiepileptic drugs than those who chose not to respond, continuing education efforts are urged to enable health care providers to counsel epileptic women about contraception.

Changes in haemostatic variables induced by oral contraceptives containing 50 micrograms or 30 micrograms oestrogen: absence of dose-dependent effect on PAI-1 activity.

Several studies have suggested a dose-response relation between the oestrogen content of oral contraceptive (OC) and the risk of both venous thrombosis and arterial disease, when oestrogen doses were higher than 50 micrograms. However, there is no clear epidemiological evidence for a decrease in thrombotic risk with formulations containing less than 50 micrograms oestrogen. Therefore, we investigated haemostatic variables in users of OC containing either 30 micrograms (35 women) or 50 micrograms (29 women) ethinyl estradiol as compared with non users (64 women) matched for age and smoking status. Mean values of antithrombin activity were significantly lower in 30 micrograms or 50 micrograms oestrogen users than in non users (96% and 98% vs 105%, respectively, p < 0.001), but they were not significantly different between the two groups of OC users. There was a significant increase in mean values of factor VII antigen in women taking either 30 micrograms or 50 micrograms oestrogen as compared with non users (96% and 101% vs 85%, respectively, p < 0.005). Although the difference between both groups of OC users was not significant, a positive linear trend in factor VII levels was observed within the 0-50 micrograms oestrogen range (p < 0.001). Mean levels of fibrinogen were slightly higher in 30 micrograms or 50 micrograms oestrogen users than in non users (2.71, 2.66 g/l vs 2.55 g/l, respectively), but there was no significant difference between the three groups.(ABSTRACT TRUNCATED AT 250 WORDS)

PIP: In Paris, France, clinicians compared data on 64 women aged 19-40 who used combined oral contraceptives (OCs) for 6-200 months with data on 64 healthy women who did not use OCs for the last two months and who were matched for age and smoking status to investigate activity of plasminogen activator inhibitor 1 (PAI-1), factor VII antigen, fibrinogen concentration, and antithrombin activity in users of OCs containing either 30 mcg or 50 mcg estrogen and in nonusers. OC users exhibited lower mean values of PAI-1 activity than nonusers (4.63-4.89 vs. 6.47 AU/ml; p 0.02). There was no dose-dependent effect of estrogen on PAI-1 activity, however. Antithrombin activity values were much lower in OC users than nonusers (96-98% vs. 105%; p 0.001). The difference between the two groups of OC users was not significant, however. The mean values of factor VII antigen in women using either 30 mcg or 50 mcg estrogen were higher than those for nonusers (96% and 101% vs. 85%, respectively; p 0.005). The difference in factor VII antigen values between the two OC groups was not significant, yet there was a positive linear trend in factor VII levels within the 0-50 mcg estrogen range (p 0.001). No significant difference in the mean fibrinogen levels between the three groups (30 mcg estrogen OC group, 50 mcg estrogen OC group, and nonusers) was observed. Hemostatic variables were not significantly different between 30 mcg estrogen OCs containing 100 mcg, 150 mcg, or 200 mcg levonorgestrel. The researchers could not conduct a valid assessment of the progestogen effect in 50 mcg estrogen OCs due to the wide range of different types of progestogens. These findings suggest an alteration of blood coagulation and fibrinolysis in OC users within the 30-50 mcg estrogen range. Estrogen appears to have a dose-dependent effect on factor VII but no significant effect on PAI-1 activity and other markers of thrombogenic risk and arterial disease risk.

Effect of oestrogen dose on whole blood platelet activation in women taking new low dose oral contraceptives.

Oral contraceptive use is known to cause changes in the haemostatic system. These changes are thought to be related to oestrogen dose and to provide a possible link between the increased risk of thromboembolic disease known to occur in women taking oestrogen containing oral contraceptives. This study measured whole blood platelet activation, serially, in women taking oral contraceptives containing 20 micrograms and 30 micrograms ethinyloestradiol combined with desogestrel. Increased levels of ADP and arachidonic acid induced aggregation were observed in women taking the 30 micrograms ethinyloestradiol combination. Platelet release of beta-thromboglobulin (beta TG) was also significantly increased. Increased collagen induced aggregation was observed but this failed to reach statistical significance for the individual treatment groups. In women taking the 20 micrograms ethinyloestradiol combination, a significant increase was only observed when platelets were stimulated with arachidonic acid. Platelet factor 4 (PF4) levels were unchanged in both groups. Significantly higher levels of beta TG were observed in women taking the 30 micrograms ethinyloestradiol combination compared with women taking the 20 micrograms ethinyloestradiol combination. These results show that oral contraceptive use is associated with platelet activation. Women taking the 20 micrograms ethinyloestradiol combination show less changes in platelet activation than women taking the 30 micrograms ethinyloestradiol combination. This lower dose pill may therefore be particularly suitable for high risk women wishing to use oral contraception.

PIP: To evaluate the effects of low-dose oral contraceptives (OCs) on platelet function, hematologic measures were compared in 45 Irish women taking OCs containing 20 or 30 mcg of ethinyl estradiol as well as 150 mcg of desogestrel. Serum samples were collected before treatment and at 6, 14, and 22 weeks after OC use commenced. ADP induced whole blood platelet aggregation was significantly increased in users of OCs containing 30 mcg of ethinyl estradiol, reaching a maximum at 22 weeks, but not in users of the low-dose OC. A significant increase in collagen induced aggregation was observed when both groups of OC users were combined, but not when either was tabulated separately. Both groups showed significant increases in arachidonic acid induced aggregation. Platelet count, hematocrit, and platelet factor 4 levels were unaffected. Increased levels of beta-thromboglobulin were observed at 6, 14, and 22 weeks in the 30 mcg group; there was no significant change in the 20 mcg group. Since the low-dose 20 mcg ethinyl estradiol OC produced fewer changes in platelet activation, its use is recommended for women with risk factors for thromboembolic disease.

The actions of ethinyloestradiol on the pituitary-adrenal system of the rat.

PIP: A study assessing the effects of estrogen on the pituitary-adrenal axis in rats is reported. In a series of experiments, rats were subjected to single and multiple doses of ethinyl estradiol (EE) and injections of ACTH. Administration of single doses of EE to quiescent rats brought increased plasma and adrenal gland corticosterone concentrations and in vitro corticosteroid production. In animals stressed by ether vapor, the plasma and in vitro corticosterone values were 40% lower than in controls although adrenal corticosterone levels were higher. Treatment with 500 mcg/kg EE per day for 7 days resulted in loss of body weight and hypertrophied, hyperdemic adrenal and pituitary glands in rats sacrificed 1 day after treatment. Rats studied 9 days after treatment showed normal growth and a regression in adrenal size but not in pituitary size. Plasma corticosterone concentration was unchanged 1 day after the 7-day treatment, adrenal weight increased by 58%, and in vivo steroid production was reduced suggesting a distinct hypersecretion of ACTH. Plasma protein binding capacity for corticosterone was unchanged by the 7-day treatment. After 9 days of rest from the EE regimen, an ACTH injection restored in vitro corticosteroid production to normal levels and raised plasma and adrenal content levels in stressed rats which suggests that the adrenal gland regained its function more quickly than the pituitary. Injection with long-acting ACTH caused a 56% increase in adrenal weight and no change in pituitary weight. Adrenal activity was not changed by ACTH treatment suggesting that the adrenal gland was insensitive to an acute release of endogenous ACTH. Inhibition of the pituitary-adrenal response to stress is most likely caused by inhibition of cholesterol synthesis although lack of precursor corticoid secretion due to exhaustion must be considered as a cause. Inhibition of the stress response after extended ACTH treatment is suggested to be due to a decreased sensitivity of the adrenal cortex although a reduction of circulating cholesterol cannot be excluded as a cause.^ieng

Proceedings: Quantitation of a new serum alpha-macroglobulin in malignant disease, steroid treatment, pregnancy and normal subjects.

PIP: Alpha-macroglobulin was quantitated in patients with malignant disease, steroid treatment, pregnancy, and in normal subjects using the rocket technique of Laurell. Women treated with combined estrogen/progestogen and with mestranol and men treated with stilbesterol showed rises in alpha-macroglobulins. Those treated with norethynodrel did not, indicating that the estrogen is the responsible agent. The level increased during pregnancy and decreased sharply in the first 2 days postpartum. 30% of normal women and 10% of normal men had detectable quantities of the protein (up to 4 mg/100 ml) in their serum. 92% of patients with malignant disease had detectable levels of protein--6 mg/100 ml or higher.^ieng

Anterior pituitary sensitivity in immature female rats stimulated with gonadotrophin releasing hormone in vivo: effect of priming with oestrogen, pregnant mare serum gonadotrophin or brain lesion.

PIP: Anterior pituitary sensitivity, assessed in terms of increments in plasma luteinizing hormone (LH) and follicle stimulating hormone (FSH) concentrations to stimulation with 1 or 2 injections of gonadotrophin releasing hormone (GnRH) was investigated in 26-day-old immature female rats which had received 1 of the following priming treatments: 1) 10 mcg estradiol benzoate (EB) as a single injection on Day 23 or Day 25 or on both days, 2) 10 IU pregnant mare serum gonadotrophin (PMSG) on Day 24, 3) an electrochemical brain lesion placed in the mediobasal hypothalamus on Day 23, or 4) control animals received either vehicle alone or a sham lesion. Pituitary sensitivity assessed at 1000 hours on Day 26 after 1 or 2 injections of GnRH was enhanced to a similar degree in the 3 groups treated with EB in terms of LH (p .01). FSH increased somewhat after EB treatment. In contrast, 48 hours after the injection of PMSG pituitary sensitivity, in terms of both LH and FSH, dropped sharply (p .001). Sensitivity to 1 injection of GnRH was unchanged in lesioned rats. However, a 2nd GnRH injection administered after an hour interval induced a slightly larger LH response in control animals. In another study, rats treated with EB on Day 23 and with 1 mg progesterone at 1200 hours on Day 26, pituitary sensitivity increased at both 1400 and 1700 hours as compared with that in the Day 23 EB-treated group at 1000 hours. PMSG-treated rats maintained their state of decreased responsiveness at 1400 hours, but exhibited increased sensitivity at the time of the gonadotropin surge (1700 hours).^ieng

Sexual behaviour of neonatally castrated rats injected during infancy with oestrogen and dihydrotestosterone.

Male rats were castrated on the day of birth (day 1) and injected with either testosterone, dihydrotestosterone, a synthetic oestrogen (RU 2858 + dihydrotestosterone, or oil from days 1 to 5. The aromatizable androgen, testosterone, and RU 2858 suppressed both cyclic gonadotrophin secretion, indicated by the absence of corpora lutea from implanted ovarian grafts, and the behavioural response to oestradiol benzoate + progesterone injections in adulthood. The 5alpha-reduced androgen, dihydrotestosterone alone did not affect gonadotrophin secretion or female receptive behaviour, but like testosterone, it increased penis development in response to testosterone propionate, and this was positively correlated with copulatory efficiency, i.e. the ratio of intromission to mount frequencies. Nevertheless, ejaculation only occurred among animals that had received testosterone or RU 2858 + dihydrotestosterone. The results support the concept that during the preinatal period, neural conversion of androgens to oestrogens is important both for the suppression of female gonadotrophin secretion and behaviour patterns as well as for the organization of male behaviour patterns. The 5alpha-reduction of unsaturated C19-steriods to dihydrotestosterone in peripheral tissues is also required to complete the development of the male genital tract.

PIP: Experiments were carried out to compare the masculine sexual behavior of male rats castrated on the day of birth (Day 1) and injected with testosterone (an aromatizable androgen), dihydrotestosterone (a nonaromatizable androgen), or an estrogen. Litters of Sprague-Dawlet rats were castrated under cryoanesthesia on Day 1 and assigned to 1 of 5 treatment groups: Group 1 received 50 mcg testosterone, Group 2 received 50 mcg dihydrotestosterone, Group 3 received RU 2858, Group 4 received 50 mcg dihydrotestosterone plus 1 mcg RU 2858, and Group 5 was given .05 ml oil. During Week 5 an ovary from a prepubertal female rat was transplanted under the left kidney capsule of each rat. Each rat was given 20 mcg estradiol benzoate per kg followed by .5 mg progesterone. Sexual receptivity was assessed. The aromatizable androgen, testosterone, and RU 2858 suppressed both cyclic gonadotropin secretion and the behavioral response to estradiol benzoate plus progesterone injections. Gonadotropin secretion or female receptive behavior was unaffected by dihydrotestosterone, but it increased penis development in response to testosterone propionate which was positively correlated with copulatory efficiency. Ejaculation only occurred in those animals that had recived testosterone or RU 2858 plus dihydrotestosteorne. During the perinatal period natural conversion of androgens to estrogens is needed for the suppression of gonadotropin secretion and behavior and for the development of male behavior patterns.

Effects of neomycin on the biliary excretion and enterohepatic circulation of mestranol and 17beta-oestradiol.

PIP: The continued circulation of free steroids depends on their resorption from the gut following the hydrolysis of biliary conjugates. In this study, the bile duct of female Wistar albino rats was cannulated. Animals receiving labeled steroids or labeled bile intraductally also had the duodenum fitted with a cannula connected with a dosing syringe. In neomycin-treated rats, recirculation was impaired up to 50%. The deconjugation of mestranol and estradiol biliary conjugates was shown in vitro uponiincubation with rat caecal microorganisms, and the inhibition of such hydrolysis by neomycin was observed in vitro. Neomycin pretreatment reduced the biliary excretion of mestranol and estradiol after intraductal administration. It was thought that suppression of the gut microflora by neomycin was a major factor in the impairment of the intrahepatic circulation of mestranol and estradiol metabolites. This effect may be important regarding the half-life of estrogenic compounds of the contraceptive pill.^ieng

Developmental correlation of higher levels of estrogen binding by macromolecules in rat liver supernatant and of increases in plasma renin substrate levels after estrogen administration.

PIP: Developmental correlation of higher levels of estrogen binding by macromolecules in rat liver supernatant and of increases in plasma renin substrate levels after estrogen administration is reported. Gel filtration columns were used to separate bound from free radioactivity in studying binding of radioactive estradiol to tissue supernatants. The liver of the prepubescent female rat has less estrogen-specific binding macromolecules than the adult (p less than .01). This difference in quantity was maintained when binding activities were partially purified by precipitation with ammonium sulfate at 30% saturation. After administration in vivo of 100 mcg of ethinyl estradiol (sc), plasma renin substrate (PRS) levels increased 167% above control in the adult female rat (p less than .05). The corresponding increase was only 15% in the prepubescent rat. In contrast, renin substrate levels were significantly increased in both the prepubescent and adult by administration of 4 mg/kg of dexamethasone (p less than .05). The marked increase in the amount of estrogen binding and PRS responsiveness to estrogen administration with sexual maturation indicates that the estrogen-binding protein may be an estrogen receptor involved in modulating synthesis of at least 1 plasma protein.^ieng

Effect of long-term therapy with an oral contraceptive on some aspects of hepatic lipid metabolism in vitro.

PIP: The effect of Enovid (7.5 mg/kg of food) for both short and long periods of therapy in female Blue Spruce Farm rats was studied. Hepatic release of triglyceride and of cholesterol was measured. Tests were made in vitro. Treatment periods were 4 days of 1 year. The short-term treated group ingested 315 mcg/kg body weight of Enovid daily; the long-term treated group ingested 375 mcg/kg of body weight of Enovid daily. These are about 3 times the daily dose for humans. Animal livers were removed and placed in a perfusion apparatus. The perfusion technique is described. The adrenal glands were also removed and the lipid extract, after being similarly treated in the perfusion apparatus, was tested for cholesterol. Body weights of animals were reduced significantly (p less than .05) by both short- and long-term treatment. The amounts of food consumed were reduced only in the short-term tests. In neither group was change in liver weight found. Bile production was reduced 50% in rats treated 4 days but in those treated 1 year with Enovid no effect was noted when compared with controls. However, in both control and test animals, production of bile after 1 year was reduced by 70-75% as compared with younger animals. Perfusion flow rate in rats treated with Enovid for 1 year was significantly faster (p less than .02) than through livers in the control group. Glucose release was reduced by both short- and long-term Enovid therapy. Reduced food intake may have caused this effect in the short-term therapy. In the group treated with Enovid for 1 year, release of cholesterol and triglycerides into the perfusate was reduced 72 and 38%, respectively. This effect was not observed in the 4-day treated animals. Enovid had no effect on the weights of livers or on the concentration of either triglycerides or cholesterol in hepatic tissue after either form of therapy. No elevation of serum triglyceride was found. A 30% decrease in serum cholesterol was found after 4 days of Enovid therapy. However, in those treated 1 year, and in controls, there was a 100% increase over that of younger animals. Enovid had no effect on the weights of adrenal glands. Total sterol content of adrenal glands from animals treated 1 year was decreased significantly (p less than .05) but not in animals treated only 4 days. Results obtained may be attributed to the metabolic effects of the individual components of Enovid. Further experiments are in progress to examine the effects of each component on hepatic triglyceride transport. Each Enovid tablet contained 5 mg norethynodrel and .075 mg of mestranol.^ieng

Effects of oestrogen and androgen on the sexual behaviour responses of the ovariectomized ewe.

PIP: The secretion of gonadal steroid hormones that stimulate sexual behavior differs between males and females in 2 respects: the hormones are chemically different, estrogen and progesterone on 1 hand and androgen on the other, and their pattern of release into the blood stream differs according to the sex of the animal. Those produced by the female are released during a limited period whereas testosterone exhibits little day-to-day variation. 12 ovariectomized Ile-de-France ewes were injected either with 50 mg of estradiol benzoate of 10 mg or testosterone propionate 48 hours after the last of 5 daily injections of 25 mg of progesterone. In both cases the experimental females exhibited normal female sexual behavior. In a 2nd experiment, the ewes were injected daily for 4 weeks either with 50 mg of estradiol benzoate or with 10 mg of testosterone propionate. In both cases male patterns of sexual behavior appeared, but more intensely with androgen than with estrogen, and simultaneously, the ewes became receptive. However, receptivity declined rapidly after a few days of estrogen treatment. This decline did not occur with androgen.^ieng

Influence of chlorpromazine on the positive and negative feed-back mechanism of oestrogens in man.

PIP: It was determined whether all the different neuroendocrine actions of estrogen are competitively antagonized by phenothiazines to test the putative analogy between the 2 types of molecular receptor. 12 men aged 22-25 years were tested on 3 days and then given 2 X 20 mg ethinylestradiol (EE) for 4 days with either 2 X 50 mg chlorpromazine (6 cases) or a placebo (6 cases). Neurohypophyseal activity and adenohypophyseal activity were tested. There was a lack of significant changes in pulse, blood pressure, body weight, and psychosexual factors. Blood alkaline phosphatases decreased in all 12 men after estrogen, and an inhibitory effect of estrogens alone was seen on blood FSH and on the 2 and 5 fraction of urinary 17-keto-steroids. The neurophysine basal level and the growth hormone peak response to hypoglycemia showed a stimulatory effect. There was no effect on FSH by chlorpromazine or on the inhibition of 17-keto-steroids due to estrogens. However, chlorpromazine lessened the neurophysine increase and abolished the facilitatory effect of estrogens on growth hormone responsibeness to hypoglycemia. In 60% of the cases TSH blood levels were undetectable and unvaried.^ieng

Long-acting hormonal contraceptives for women.

Following the development and widespread use of oral hormonal contraceptives, it became evident that alternative long-acting delivery systems would be required to improve contraceptive practice in some cultural settings where injectable or subdermal routes of administration are preferred. Nowadays, long-acting contraceptives constitute an important option in family planning services in many parts of the world. Indeed, two long-acting injectable contraceptives containing just a synthetic progestogen (depot-medroxyprogesterone acetate (DMPA) and norethisterone enantate (NET-EN)) have been in clinical practice for more than 20 years. The World Health Organization's (WHO) Special Programme of Research in Human Reproduction, in collaboration with the U.S. National Institute of Child Health and Human Development (NICHD) and universities primarily in developing countries undertook a synthesis programme aimed at producing an improved injectable preparation by developing new derivatives of known steroids. One such compound (levonorgestrel 17-butanoate) is now at the stage of Phase II clinical testing. In addition, the Special Programme has developed and improved once-a-month injectable formulations and assessed their safety and efficacy in different countries worldwide. After large scale clinical testing, at least two progestogen-estrogen combinations have reached the point of introductory trials.

PIP: A survey of recent trials of new injectable hormonal contraceptives, progestogen-only, levonorgestrel esters, and once monthly injectables, follows a brief review of all the experimental long-acting contraceptive modalities, injectables, implants, vaginal rings, and hormone-releasing IUDs. Currently medroxyprogesterone acetate (DMPA) and norethisterone enanthate (NET-EN) are being used by 7 million women. WHO is conducting dose reduction trials and studies of bioavailability in various national populations. Even though a dose of 100 mg DMPA every 3 months has been satisfactory for contraception, 150 mg is still recommended until further pharmacodynamic data are available. Some populations, notably Thais and Mexican women, have higher peaks and more rapid elimination rates of DMPA, while Chinese women show slower elimination and higher blood levels of NET-EN. Extensive studies of new synthetic esters of levonorgestrel have proceeded to Phase II clinical trials with levonorgestrel butanoate. This ester is an effective contraceptive for 3 months at 12.5 mg, or 5-6 months at a dose of 25 or 50 mg. Trials of combined estrogen and progestogen injectables once-monthly have been ongoing for 10 years. The ratio of the 2 components is as important as the amounts. 2328 women from 12 countries participated in trials of DMPA 25 mg-estradiol cypionate 5 mg, and NET-EN 50 mg-estradiol valerate 5 mg. The continuation rate was better than that for 3-monthly progestogen-only injectables, because of less irregular bleeding. A combined injectable called Cyclofem, DMPA 25 mg-estradiol cypionate is being introduced in several countries. The steadily increasing demand for long-acting injectables prompts development of better formulations.

Differential lipemic and proteinemic response to oral ethinyl estradiol and parenteral estradiol cypionate.

Oral synthetic estrogen administration to normal women has been shown to result in both a lipemic and a proteinemic response. To determine whether parenteral estrogen administration would have similar results, the effects of intramuscular depo-estradiol cypionate on serum lipids and ceruloplasmin were examined. The oral and parenteral estrogens chosen for this study are frequently used therapeutically and varying doses in the range of those commonly employed clinically were given to the experimental subjects. Following oral ethinyl estradiol (20, 50, and 100 micrograms every 12 hr) comparable and significant increases in triglyceride (73 +/- 6 to 128 +/- 10 mg/dl, p < .001), ceruloplasmin (87 +/- 4 to 188 +/- 11 mg/dl, p < .001), and HDL-cholesterol (60 +/- 2 to 74 +/- 3 mg/dl, p < .001) were observed. In contrast, despite substantial increases in serum estrogens, parenteral estrogen administration (depo-estradiol cypionate, 5 and 10 mg) failed to result in alterations in any of the measured parameters. Thus, the route and/or type of estrogen administered may determine the proteinemic and lipemic effects of estrogen in man.

PIP: This study looked at the effect of a commonly used estrogen preparation, parenteral depo-estradiol cypionate, on several metabolic parameters and compared the results of known alterations which follow oral ethinyl estradiol. The metabolic parameters studied were serum cholesterol, triglyceride (TG), ceruloplasmin, and high-density lipoprotein (HDL) cholesterol. Doses of agents used are common therapeutic ones. After oral ethinyl estradiol (20, 50, and 100 mcg every 12 hours) significant increases in TG (73+ or -6 to 128+ or -10 mg/dl, P .001), ceruloplasmin (87+ or -4 to 188+ or -11 mg/dl, P .001), and HDL-cholesterol (60+ or -2 to 74+ or -3 mg/dl, P .001) were observed. In contrast, despite substantial increases in serum estrogens, parenteral estrogen administration (depo-estradiol cypionate, 5 and 10 mg) failed to result in alterations in any of the measured parameters. Thus, the route of contraceptive agent administration may affect the proteinemic and lipemic responses in humans.

Letter: The effect of contraceptive steroids on carbohydrate metabolism.

PIP: Comments are made on a short review on the effect of contraceptive s teroids on carbohydrate metabolism which appeared previously. Intraveno us tolbutamide tolerance tests were performed on 13 normal women and 8 hypopituitary patients before and 3 and 6 months after starting on an oral contraceptive agent (OCA) containing 50 mcg mestranol and 1 mg norethindrone. The control subjects demonstrated a significant impairment of the glucose fall after tolbutamide and an enhancement of the growth hormone (GH) response to the resultant hypoglycemia while on the OCA. The pituitary-deficient patients showed no change in their glucose response to tolbutamide after starting OCA and they had minimal GH secretion as expected. 5 additional normal females on a variety of OCAs had a significant 3-fold increase in GH concentrations after moderate exercise as compared with 5 other women using other forms of contraception. The peripheral tissues of women on OCAs were exposed to higher levels of GH throughout the day. The data seemed to implicate the anterior pituitary gland, and it was postulated that the estrogen-induced secretion of GH, a potent insulin antagonist, was responsible for the effects of OCAs on carbohydrate metabolism. It is concluded through laboratory investigations that there is good evidence to support the role of estrogens alone in causing deterioration of glucose tolerance. This contradicts the review in which it was concluded that carbohydrate metabolism in younger women was unaffected by estrogens alone but will deteriorate in response to a combination of estrogen and nortestosterone progestins.^ieng

Biliary stasis after mestranol-norethindrone ingestion. Report of a case.

PIP: This case report concerns a 29-year-old caucasian woman who developed cholestatic jaundice while taking Ortho-Novum 2 mg. Biliary stasis was diagnosed from laboratory tests and gradually subsided when the medication was withdrawn. The condition was found to be reproducible when Ortho-Novum 2 mg. was given again. The patient was later given C-Quens with no return of symptoms which indicated norethindrone was the cholestatic agent.^ieng