Acute phase protein in serum of women using hormonal contraceptives.

Acute phase protein precipitating sonatic C-polysaccharide of pneumococci appears in serum of women under treatment with hormonal contraceptives in a significantly higher number of cases when compared with control groups. The summarized results of three preliminary studies show that in 80 control serums there were four positive specimens (5 percent) and in 80 serums from women using hormonal contraceptives there were 58 positive specimens (72.5 percent).

Oral contraceptives as substrates and inhibitors for human cytosolic SULTs.

Cytosolic sulfotransferases (SULTs) in mammals are involved in the biotransformation and homeostasis of various endogenous and xenobiotic compounds. The current study aimed to examine the sulfation of contraceptive compounds by various human cytosolic SULTs and to investigate the inhibitory effects and mode of action of these compounds on the sulfation of 17beta-estradiol, a major endogenous estrogen. A systematic study using all eleven known human cytosolic SULTs revealed the differential substrate specificity of these enzymes for the eight representative contraceptive compounds and two endogenous estrogens (estrone and 17beta-estradiol) tested as substrates. Activity data showed that SULT1A1 displayed the strongest activity toward 17alpha-ethynylestradiol. Kinetic studies revealed that the V (max) value of the sulfation of 17alpha-ethynylestradiol by SULT1A1 was 1.64 times that of the sulfation of 17beta-estradiol, while the K (m) values were almost equal for the two compounds. The inhibitory effects of three contraceptive compounds on the sulfation of 17beta-estradiol by SULT1A1 were examined. IC(50) values determined were 0.193, 1.84, and 2.98 mM, respectively, for 19-norethindrone acetate, ethynodiol diacetate and mifepristone. Kinetic analyses indicated that the mechanism underlying the inhibition by these contraceptives is of a mixed noncompetitive type. Metabolic labeling experiments confirmed the sulfation of contraceptive compounds and the release of their sulfated derivatives by HepG2 human hepatoma cells. Collectively, the results obtained suggest a role of sulfation in the metabolism of contraceptive compounds in vivo. Moreover, in view of their inhibitory effects on the sulfation of 17beta-estradiol, these compounds may potentially act to disrupt the homeostasis of endogenous estrogens.

Androgen control of an inhibitory modulator of phosphodiesterase in rat epididymis and prostate.

When rats were injected with the progestin ethynodiol diacetate in doses that suppressed spermatogenesis and the growth of accessory sex glands, the level of phosphodiesterase in epididymal and prostate tissues increased 5- to 10-fold. This increase was prevented by concurrent administration of testosterone propionate. A similar increase in phosphodiesterase activity was observed in the epididymides and prostates of castrated animals, with reversal by treatment with androgen. In immature rats approaching puberty, the phosphodiesterase activity in epididymis and prostate increased while the blood testosterone level remained low; as the testosterone level rose with the onset of puberty, the phosphodiesterase activity decreased. Incubation of enzymically active extracts of accessory tissues from castrated rats with heat-treated extracts of the corresponding normal tissues resulted in strong inhibition of the initially high phosphodiesterase activity. The addition of heat-treated extracts of accessory glands from castrated rats to enzymically active extracts of the corresponding tissues from normal rats resulted in a marked elevation of their phosphodiesterase activities. Of the two heat-stable modulators, the inhibitor factor was dialyzable. The dependence of this factor on testosterone suggests a mechanism of action by which the steroid hormone, by inducing the production in its target tissues of an inhibitory modulator of phosphodiesterase, controls the maintenance of functional levels of cAMP.

Contribution of functional groups of 19-nor-progestogens to binding to progesterone and estradiol-17beta receptors in rabbit uterus.

The structural elements of 19-norprogestogens which may be essential for binding to progesterone and estradiol-17beta(E2) receptors were investigated in the rabbit uterine cytosol. The kinetic study showed that 19-nor-progestogens are competitive inhibitors of progesterone-receptor (8S) binding and E2-receptor binding. The affinities of steroids for the progesterone receptor were as follows: norethindrone (Ki of 2.3 X 10(-9)M) greater than 5alpha-dihydronorethindrone greater than norethindrone acetate greater than lynestrenol greater than 17alpha-ethynyl-estra-4-ene-3beta, 17beta-diol greater than ethynodiol diacetate (Ki of 1.3 X 10(-7) M). The affinities of steroids for the E2 receptor were as follows: ethynodiol diacetate (Ki of 1.3 X 10(-7)M) greater than 17alpha-ethynyl-estra-4-ene-3beta, 17beta-diol greater than norethindrone acetate greater than norethindrone greater than 5alpha-dihydronorethindrone greater than lynestrenol (Ki of 8.4 X 10(-7)M). The results indicate that 3-ketone and 17beta-hydroxyl groups, and the plane of ring A/B of 19-norprogestogen are important for binding to the progesterone receptor. The affinities of 19-nor-progestogens for the E2 receptor were very weak. Their affinities for the E2 receptor increased with addition of acetate or hydroxyl groups at the 3beta and 17beta positions, and were decreased by the elimination of a 3 oxygen function or the reduction of ring A.

Effects of oral contraceptive agents on copper and zinc balance in young women.

Young adult women were selected from a group of volunteers on the basis of present and past use of oral contraceptive agents. Subjects were either those who had never used oral contraceptives or those who had used an agent for a minimum of two menstrual cycles. All subjects were fed diets that provided 2.02 to 2.11 mg of copper and 9.15 to 9.10 mg of zinc daily in an 18-day controlled metabolic study. Both oral contraceptive users and nonusers were in negative zinc balance and zinc excretion as not different (P > 0.05) for the groups. Oral contraceptives did not alter (P > 0.05) the metabolic balance of copper; the mean copper retention values were -0.16 mg/day for the nonusers and -0.13 mg/day for the users of oral contraceptives. As anticipated from reports in the literature, the plasma copper levels were higher (P < 0.01) in the oral contraceptive group, but the levels of zinc were not different. Although blood levels of zinc and copper are usually changed with the use of oral contraceptives, the retention of the two nutrients appears not to be affected.

PIP: Young adult women were selected from a group of volunteers on the basis of present and past use of (OCs) oral contraceptives. Subjects were either those who had never used OCs or those who had used an agent for a minimum of 2 menstrual cycles. All subjects were fed diets which provided 2.02-2.11 mg of copper and 9.15-9.10 mg of zinc daily in an 18-day controlled metabolic study. Both OC users and nonusers were in negative zinc balance and zinc excretion was not different (P 0.05) for the groups. OCs did not alter the metabolic balance of copper (P 0.05); the mean copper retention values were -0.16 mg/day for the nonusers and -0.13 mg/day for OC users. As anticipated from reports in the literature, the plasma copper levels were higher (P 0.01) in the OC group, but the levels of zinc were not different. Although blood levels of zinc and copper are usually changed with the use of OCs, the retention of the 2 nutrients appears not to be affected.

Effect of ethynodiol diacetate and mestranol on serum folic acid and vitamin B12 levels and on tryptophan metabolism in baboons.

The effect of an oral contraceptive agent on serum folate and vitamin B12 levels and on some aspects of tryptophan metabolism in the baboon has been investigated. Folate levels were not affected by this treatment. Vitamin B12 levels were lowered during oral contraceptive treatment (P smaller than 0.1) and increased during a 30-day period of pyridoxine supplementation (P smaller than 0.01). Trends indicating deterioration of glucose tolerance and abnormal tryptophan metabolism were also observed. Definite conclusions concerning the effect of oral contraceptive agents on nutritive parameters could not be made. This is in keeping with the vast volume of literature published on the effects of oral contraceptive agents in humans and other species. It is concluded that perhaps, due to the biologically variable responses to oral contraceptive agents by individuals of any species, studies of serum levels of vitamins and other superficial biochemical parameters. Further, we conclude that the baboon could be a useful model for the study of endocrine influences on nutritive parameters, first because endocrine parameters resemble those of humans and second, because experimental protocols can be rigidly adhered to.

Effect of oral contraceptive agents on vitamin nutrition status.

Effects of low estrogen combination type oral contraceptives on some of the biochemical parameters used for assessing vitamin nutritional status were investigated in a group of women who had used the pill for 6 to 12 months. Another group of women was examined initially and then at one or more points of time within the first 6 months of treatment. Following changes were observed in women treated with oral contraceptives: 1) increased excretion of kynurenic acid and xanthurenic acid following tryptophan load; 2) increased EGOT activity and also an increase in vitro stimulation of EGOT with added PALP; 3) increased plasma vitamin A levels; 4) fall in erythrocyte folate levels; 5) fall in erythrocyte transketolase activity with no change in vitro stimulation with TPP; and 6) fall in erythrocyte riboflavin concentration associated with a decrease in erythrocyte glutathione reductase activity and increase in vitro stimulation with FAD. Most of these changes were observed during the first few cycles of oral contraceptive treatment.

Metabolic effects of oral contraceptives in monkeys fed on adequate and low protein diets.

PIP: Rhesus monkeys were given either high estrogen, low progesterone or low estrogen-high progesterone oral contraceptive agents (OCAs) and maintained on either a 16% protein diet or a 4% protein diet. Both OCAs created a small but significant fall in hemoglobin and serum protein. On both diets, OCAs led to a gradual elevation of serum alkaline phosphatase and serum glutamic oxalic transaminase (SGOT) activities. There were no histologic liver lesions. On the low protein diet, glucose tolerance was impaired between cycles 10 and 11, and on the high protein diet between cycles 16 and 20. Vitamins B-12, A, and thiamine tended to be lower in OCA animals. The conclusion of these experiments is that the hazard of using OCAs is not exaggerated by protein deficiency.^ieng

Raised serum copper and caeruloplasmin levels in subjects taking oral contraceptives.

A marked increase in the serum copper and caeruloplasmin levels in 25 female subjects taking oral contraceptives is noted. The correlation between the copper and caeruloplasmin concentrations, together with the finding of normal urine copper levels in 10 of the subjects, suggests that the increased serum copper is due to increased caeruloplasmin synthesis.

Urinary excretion of quinolinic acid and other tryptophan metabolites after deoxypyridoxine or oral contraceptive administration.

PIP: The urinary excretion of quinolinic acid and some other tryptophan metabolites has been determined after a 2-g L-tryptophan load in a deoxypyridoxine-treated subject, in 20 women receiving estrogen-containing oral contraceptives, in and a control group of 12 women taking no steroids. In both situations there were increases in the excretion of quinolinic acid and 3-hydroxyanthanillic acid, as well as 3-hydroxykynurenine and xanthurenic acid. These changes were reversed by pyridoxine administration and are considered to reflect an inhibitory effect of estrogens and deoxypyridoxine on vitamin B6 dependent enzymes. Although the principal enzyme of the tryptophan-nicotinic acid ribonucleotide pathway affected by imparied pyridoxal phosphate function is kynureninase, the present results are consistent with the existence of an unidentified enzyme beyond the formation of 3 hydroxyanthranilic acid requires the coenzyme.^ieng

Effects of oral contraceptives on lipoprotein lipids: a prospective study.

One hundred sixty-nine healthy women, aged 17 to 29 years, nonsmokers or light smokers (fewer than ten cigarettes per day), were assigned randomly to take one of five oral contraceptives: 1) 100 micrograms mestranol plus 0.5 mg ethynodiol diacetate (100 M + 0.5 ED); 2) 100 micrograms mestranol plus 1.0 mg ethynodiol diacetate (100 M + 1.0 ED); 3) 50 micrograms ethinyl estradiol plus 1.0 mg ethynodiol diacetate (50 EE + 1.0 ED); 4) 30 micrograms ethinyl estradiol plus 2.0 mg ethynodiol diacetate (30 EE + 2.0 ED); or 5) 30 micrograms ethinyl estradiol plus 0.15 mg levonorgestrel (30 EE + 0.15 NG). One hundred forty-seven women completed the study. When assessed for within-group differences, all preparation caused a statistically significant increase in total triglyceride (from 17.0 to 46.4 mg/dL), total cholesterol (from 6.3 to 24.4 mg/dL), and low-density lipoprotein (LDL) cholesterol (from 7.0 to 10.3 mg/dL). Effects on high-density lipoprotein (HDL) cholesterol varied widely. The product 100 M + 0.5 ED markedly increased (9.9 mg/dL) HDL cholesterol. Neither 100 M + 1.0 ED nor 50 EE + 1.0 ED altered HDL cholesterol levels, whereas both preparations containing 30 micrograms estrogen showed decreases: the preparation containing 2.0 mg ethynodiol diacetate lowered HDL cholesterol by 3.6 mg/dL and that containing 0.15 mg levonorgestrel lowered it by 6.9 mg/dL. Specific between-group comparisons revealed no statistically significant differences between differing amounts of estrogen (50 EE + 1.0 ED versus 100 M + 1.0 ED).(ABSTRACT TRUNCATED AT 250 WORDS)

Glucose metabolism during the menstrual cycle. Glucose tolerance during the one cycle and under the effect of ovulatory suppressants.

PIP: To investigate possible diabetogenic effects of endogenous and exogenous hormones, an intravenous glucose tolerance test was administered to 3 groups of women: those with a normal cycle (15), those on Sequalan sequential therapy (12), or those on progestational therapy (13). None had received oral contraceptives previously. The test was performed for every woman at the estrogen peak (day 13, 14, or 15) and at the progesterone peak (day 23, 24, or 25) of a 28-day cycle. For each of the three groups there was no significant glucose metabolism difference between the two phases of the cycle. Estrogens could still reduce glucose tolerance if the luteal peak of urinary estrogen secretion, which occurs in most cycles, is characterized by the presence of nonbiologic estrogen precursors for estriol. Glucose tolerance can be investigated at any part of the cycle.^ieng

The effect of ampicillin on oral contraceptive effectiveness.

The effectiveness of oral contraceptives with concomitant use of ampicillin has been questioned. Eleven volunteer subjects were studied during 2 consecutive menstrual cycles; they were given Demulen with placebo or ampicillin during each of the cycles. Serum estradiol (E2), progesterone (P4), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) were evaluated on days 13 and 19 of each cycle. Testosterone-binding globulin (TeBG) capacity and ferning of cervical mucus were evaluated on day 13 of each cycle. Subjective symptoms were recorded. All cycles appeared to be anovulatory. Steroid suppression was present in 10 of 11 patients during both cycles. Estrogen-dependent TeBG was not affected by ampicillin. It is possible that cervical mucus ferning was improved by concurrent ampicillin administration. Ampicillin appears unlikely to diminish the effectiveness of the oral contraceptive studied.

Studies on dopamine turnover in ovariectomized or hypophysectomized female rats. Effects of 17 beta-estradiol benzoate, ethynodioldiacetate and ovine prolactin.

The effects of 17 beta-estradiol benzoate (EB), ethynodioldiacetate and ovine prolactin on dopamine (DA) turnover have been studied. As an index for a change in turnover, differences in DA depletion following tyrosine hydroxylase inhibition with alpha-methyl-tyrosine methylester were observed. DA was measured by means of mass fragmentographical analysis in the rat median eminence, the olfactory tubercle and the striatum. The actions of ethynodioldiacetate and ovine prolactin on DA turnover in various subregions of the rat median eminence were analysed by quantitative microfluorimetry. Repeated injections of EB to ovariectomized rats resulted in a significant increase of DA turnover in the median eminence. Administration of ethynodioldiacetate to ovariectomized rats almost significantly increased DA turnover in the olfactory tubercle. In the median eminence DA turnover was significantly increased only in the lateral palisade zone. In male hypophysectomized rats ovine prolactin increased DA turnover in both the lateral and the medial palisade zone of the median eminence. The results support the involvement of DA neurons in the control of prolactin and luteinizing hormone secretion. It is suggested that the tubero-infundibular DA neurons are involved in mediating the central inhibitory feedback actions of prolactin and gonadal steroids on prolactin and LH secretion, respectively.

Carbohydrate and lipid metabolic studies before and after one year of treatment with ethynodiol diacetate in "normal" women.

A 3-hour oral glucose tolerance test was conducted in "normal" women before and after 12 months of contraceptive treatment with daily 0.25-mg ethynodiol diacetate tablets. There was no significant change in the subjects' weight between the two tests. All but the 3-hour individual group glucose values were significantly elevated at the 1-year test, and 16.7% of the tolerance curves became "borderline abnormal". The group plasma insulin values for the 0.5-, 1-, and 2-hour samples during the 1-year test were also significantly elevated. There was a significant decrease in the group fasting triglyceride levels but no change in the fasting cholesterol levels. These data suggest that some of the 19-norprogestogens may adversely affect carbohydrate metabolism.

The effect of combination and low dose progestogen oral contraceptives on serum lipids.

Fasting serum triglyceride, free fatty acid, phospholipid, and cholesterol levels were determined in women who were taking either combination or low dose progestogen oral contraceptives. The contraceptives used were: ethynodiol diacetate and ethynyl estradiol; norgestrel and ethynyl estradiol; norgestrel; and megestrol acetate. Serum lipids significantly increased in women taking combination pills. They significantly decreased in patients taking norgestrel, while they remained almost unchanged in patients taking megestrol acetate. This decrease varied with the type of progestogen used in continuous low dosage therapy. The confusion when interpreting the results from different reports on the effects of oral contraceptives on serum lipids will be reduced if proper attention is given to the types of steroid used.

The effect of oral contraceptives on serum lipids.

PIP: In the latter part of pregnancy total lipids, cholesterol, triglycerides, phospholipids, and free fatty acids increase progressively. A study involving 375 women aged 19-30 taking various oral contraceptives, both combined and sequential, showed a mean value of 712 mg/100 ml blood of total lipids as contrasted with 596 mg/100 ml for controls. Serum triglycerides also showed marked elevation -- 122 mg/100 ml as compared with 70 mg/100 ml for controls. However, there was no significant change in the level of cholesterol. There was no significant difference between the combined and sequential users or between users of pills of different steroid composition. Total serum lipids and triglyceride levels were similar to those seen in the last 2 trimesters of pregnancy; however, in pregnancy the cholesterol level also increases. Estrogens are known to elevate serum triglycerides, and in hypercholesteremic subjects, estrogen administration lowers serum cholesterol concentrations.^ieng

The relative expressed estrogenicity of oral contraceptives.

PIP: Total effective estrogenicity of several human oral contraceptives was investigated through a method determining uterine weight in immature rats. The preparations, normally given to human females in tablet, form were Norinyl-1 (.05 mg mestranol, 1mg norethindrone), Norinyl-2 (.10 mg mestranol, 2 mg norethindrone), Ovulen (.10 mg mestranol, 1mg ethynodiol diacetate), Enovid-E (.10 mg mestranol, 2.5 mg norethynodrel), and Enovid-5 (.075 mg mestranol, 5mg norethynodrel). Each products was administered in total doses of .0025, .005, and .01 of a tablet. Each dose had 3 trials, approximately 10 mice to a trial. Each mouse was given the compound in .2 ml aqueous vehicle orally, daily for a 3-day period so that the sum of the daily doses equaled 1 of the tablet fractions above. Autopsy, on Day 4, discovered the ratio of mg uterus/gm body weight. A .0025 fraction of a tablet of Norinyl-1 produced a mean uterine ratio of 1.79, while control (untreated) mice had a mean ratio of 1.06. Norinyl-2, Ovulen, Enovid-5 and Enovid-E, judging from the corresponding increases in ratios from the control ratio, were found to be 2.1, 2.7, 6, and 8 times, respectively, more estrogenic than Norinyl-1. The relatively high estrogenicity of the Enovids is probably due to the conversion of norethynodrel to estrogenic compounds.^ieng

Ocular effects of oral contraceptives. I. Studies in the dog.

The administration of two oral contraceptives to female dogs for 5 years did not produce ocular lesions. Corneal and lenticular opacities occurred with equal frequency in control and treated groups, and fundic lesions, including papilledema, venous dilatation, and venous or arterial retinal thrombosis, were not produced by doses of Enovid-E or Ovulen 1, 10, and 25 times the human dose.