Studies on the role of intestinal bacteria in metabolism of synthetic and natural steroid hormones

Administration of antimicrobial agents to subjects taking oral contraceptives has been reported to lead to contraceptive failure and subsequent pregnancy. In women taking oral contraceptives antimicrobial agents could have an effect on both endogenous hormone levels and on the metabolism of the exogenously administered steroids. To investigate these possibilities, antimicrobial agents were administered for short periods to normal women taking various steroid drugs: Megestrol acetate (MA), medroxyprogesterone acetate (MPA), norethisterone (NET), a combination of NET and ethinylestradiol (EE) or a combination of lynestrenol and EE. During ampicillin administration the 24-h morning plasma concentrations of MA, MPA and NET were increased compared to the control values. In the MA and MPA experiments the afternoon values were determined and also found to be increased. In the subjects taking oral contraceptives plasma EE concentration showed a tendency to decrease during ampicillin administration on the third, fourth or fifth morning of ampicillin administration, but was never lower than the pretreatment values. In other experiments plasma estrone (E1) and estradiol (E2), urinary total E1, E2 and estriol (E3) and fecal unconjugated and conjugated E1, E2 or E3 were determined by RIA before, during and after administration of oxytetracycline (2 X 500 mg/day for 5 days) to 5 young male subjects. Furthermore urinary and fecal estrogens were determined in 1 male subject after administration of erythromycin for 6 days and in 2 normally menstruating women after tetracycline and trimethoprim administration, respectively. During treatment with antimicrobial drugs an increase in the excretion of fecal conjugated and, with the exception of the oxytetracycline experiments, also of unconjugated estrogens paralleled a decrease in urinary estrogen excretion, especially for E2 and E3. In both urine and feces the E1/E2 and E1 + E2/E3 ratios increased due to diminished reductive metabolism of estrogens in the gut. No significant effects on plasma unconjugated estrogen concentrations were observed. The results suggest that the intestinal bacterial flora plays a significant role in estrogen metabolism. However, further studies are necessary, because our results do not explain why administration of antibiotics may cause contraceptive failure.     

Hormone levels and anogenital swelling of female chimpanzees as a function of estrogen dosage in a combined oral contraceptive.

A combined oral contraceptive consisting of ethinyl estradiol (EE2) in three dosages (50, 100, and 400 micrograms) and norethindrone (0.5 mg) was given to female chimpanzees to determine the effect on endogenous sex hormone levels and anogenital swelling. Serum levels of EE2 increased with increasing dosages of EE2, estradiol decreased, and luteinizing hormone, progesterone and testosterone were maintained at approximately midfollicular phase levels. Urinary levels of EE2 glucuronide increased with the increasing dosages of EE2, whereas estrone and pregnanediol glucuronide were essentially undetectable. The cyclic increase in female anogenital swelling was abolished when the norethindrone was combined with 50 micrograms of EE2 and relatively constant and low levels of swelling were recorded. Relatively constant but successively higher levels of swelling were recorded when the norethindrone was combined with the higher dosages of EE2. These effects of oral contraceptives on female genital tissues are relevant to our laboratory studies of sexual behavior in chimpanzees given oral contraceptives and could also have implications for women taking oral contraceptives.     

Excretion of non-conjugated androstenedione and testosterone in human urine.

A method for the measurement of unconjugated testosterone and androstenedione in human urine is described. The method uses chromatographic separation followed by radioimmunoassay and has been examined for reliability. The mean 24-hour excretion of androstenedione by adult male subjects was 2.5 mug and of testosterone was 0.8 mug. For women, the mean excretion was 2.9 mug of androstenedione and 0.25 mug of testosterone. In pregnancy, androstenedione excretion was occasionally elevated above the normal range, but testosterone excretion was quite commonly increased. Some hirsute subjects exhibited an increase in androstenedione excretion, which was decreased by administration of dexamethasone. The results suggest that the amount of unconjugated testosterone in urine is not a direct reflection of the plasma free testosterone, but urinary androstenedione may be a useful reflection of plasma androstenedione levels.     

Hormone levels in serum and seminal plasma of men with different types of azoospermia

Hormone concentrations in the serum and seminal plasma of 15 normozoospermic, 17 excretory azoospermic and 14 secretory azoospermic men were measured. The results indicate that: (a) serum FSH and LH levels are markedly elevated in secretory azoospermia, as compared with excretory azoospermia and normozoospermia; (b) serum 17 alpha-hydroxyprogesterone levels are somewhat raised in secretory azoospermia as compared with excretory azoospermia and normozoospermia; (c) serum testosterone levels are lower in both types of azoospermia with respect to normozoospermia; (d) in secretory azoospermia the oestradiol serum levels are relatively high and dihydrotestosterone serum levels relatively low, whereas the serum levels of these hormones in excretory azoospermia are similar to those in normozoospermic men; (e) in the seminal plasma of azoospermic patients the levels of prolactin, progesterone, testosterone, dihydrotestosterone and oestradiol were depressed, but only dihydrotestosterone levels could be of value in differentiating types of azoospermia because they are lower in secretory azoospermia. We suggest that the measurement of FSH, LH, 17 alpha-hydroxyprogesterone, dihydrotestosterone and oestradiol in serum and dihydrotestosterone in seminal plasma may be used in the differential diagnosis between secretory and excretory azoospermia when invasive tests are unavailable.     

Oral contraceptive use in older women and fatal myocardial infarction.

A previous study of women who had died from myocardial infarction and of a control group of women matched with them for age suggested a fivefold increase in the risk of death from myocardial infarction among users of oral contraceptive aged 40-44 years compared with women not using such preparations. Only a small proportion of women in the infarction and control groups had used oral contraceptives, however, so the margin of error was wide. We therefore investigated a further 54 women in this age group who died from myocardial infarction and compared their oral contraceptive histories with those of age-matched, living controls. Combination of the findings from the present investigation with the previous results have enabled a revised estimate of a threefold increase in risk to be made. Although this risk estimate is similar to that previously shown for a younger age group, the total mortality attributable to complications associated with the use of oral contraceptives remained considerably greater among women over the age of 40.     

Sexual desire, sexual arousal and hormonal differences in premenopausal US and Dutch women with and without low sexual desire

The interaction between women's hormonal condition and subjective, physiological, and behavioral indices of desire or arousal remains only partially explored, in spite of frequent reports from women about problems with a lack of sexual desire. The present study recruited premenopausal women at two sites, one in the United States and the other in the Netherlands, and incorporated various measures of acute changes in sexual desire and arousal. A sample of 46 women who met criteria for Hypoactive Sexual Desire Disorder (HSDD) was compared to 47 women who experienced no sexual problems (SF). Half of each group used oral contraceptives (OCs). The specific goal was to investigate whether there is a relationship between women's hormone levels and their genital and subjective sexual responsiveness. Background demographics and health variables, including oral contraceptive (OC) use, were recorded and hormones (total testosterone (T), free testosterone (FT), SHBG, and estradiol) were analyzed along with vaginal pulse amplitude and self-report measures of desire and arousal in response to sexual fantasy, visual sexual stimuli, and photos of men's faces. Self-reported arousal and desire were lower in the HSDD than the SF group, but only for women who were not using oral contraceptives. Relationships between hormones and sexual function differed depending on whether a woman was HSDD or not. In line with prior literature, FT was positively associated with physiological and subjective sexual arousal in the SF group. The HSDD women demonstrated the opposite pattern, in that FT was negatively associated with subjective sexual responsiveness. The findings suggest a possible alternative relationship between hormones and sexual responsiveness in women with HSDD who have characteristics similar to those in the present study.     

Serum, urinary, and salivary nitric oxide in rheumatoid arthritis: complexities of interpreting nitric oxide measures

Nitric oxide (NO) may play important roles in rheumatoid arthritis (RA). RA is an inflammatory disease involving joints and other systems including salivary glands. To assess NO production in RA patients, we compared levels of serum, urine, and salivary nitrite and nitrate (NOx) in patients with RA and normal subjects, and we examined the relationships of these measures to disease activity. Serum, urine, and NOx levels as well as renal creatinine, NOx clearance and fractional excretion rates were compared in 25 RA patients and 20 age- and gender-matched healthy controls. Subjects were hospitalized for 3 days and placed on a NOxrestricted diet. NOx was assayed using nitrate reductase and the Griess reagent. RA activity was assessed using standard clinical and laboratory measures. While consuming a restricted diet for 3 days to eliminate the effects of oral intake of NOx, 24 hour urinary NOx excretion decreased in both RA patients and healthy controls. Urine NOx levels at all time points were not significantly different between RA patients and normal subjects. Serum NOx levels also decreased during the 3 days of NOx restriction, but RA patients had higher serum NOx levels at all time points compared with the control group. Likewise, serum NOx/creatinine ratios were higher in RA patients than in controls. Although basal salivary flow rate and tear flow were lower in RA patients, salivary NOx levels did not differ between normal and RA subjects. While renal creatinine clearance was not different between the two groups, we found that RA patients had lower renal NOx clearance and lower renal NOx fractional excretion. After correction of p values for multiple comparisons, there were no significant relationships for the RA group between measures of disease activity and the urinary NOx, serum NOx, or urinary NOx clearance. Despite interest in the use of NO as a marker of disease activity, alterations in renal NOx clearance and fractional excretion in RA make it difficult to assess in vivo NO production even with strict dietary restriction of NOx intake.     

Studies of Carbohydrate and Lipid Metabolism in Women Developing Hypertension on Oral Contraceptives

Metabolic studies in 100 women developing hypertension on combined oestrogen-progestogen oral contraceptives have been compared with similar studies in normotensive women on oral contraceptives, matched for age and duration of contraceptive use, and in women not taking contraceptives.The metabolic changes known to be induced by oral contraceptives—impaired glucose tolerance, elevated blood pyruvate levels, and raised serum lipid concentrations—were found to be exaggerated in the matched hypertensive group, largely due to pronounced abnormalities in 33 subjects with diastolic blood pressures over 110 mm Hg.Women developing severe hypertension were older, more obese, and of higher parity than those with mild hypertension and there was a high incidence of previous toxaemia of pregnancy in the hypertensive group.The results show that in women on oral contraceptives changes in blood pressure and in metabolic functions tend to be correlated with one another, and are consistent with the hypothesis that oral contraception induces a primary biochemical effect whose expression in the individual is determined by intrinsic factors including genetic constitution, age, weight, and parity.     

Trace element status (Se, Cu, Zn) in healthy portuguese subjects of Lisbon population

Serum levels of selenium, copper, and zinc were systematically determined in healthy subjects of the Lisbon population. The sample consisted of 183 blood donors of both genders who were divided into two age groups: 20–44 and 45–70 yr of age; relationships with gender, age, the lipid profile, and tobacco consumption were investigated. In the female group, the intake of oral contraceptives and pregnancy were considered for the youngest women, and hormonal replacement therapy (HRT) was taken into account for the oldest ones. Serum concentrations of these elements were in the same range as those found for populations of other European countries. Differences between genders were observed for the three elements studied, with serum selenium and zinc concentrations higher in men and copper levels higher in women. Age-dependent differences were found for selenium: The oldest subjects (regardless gender) presented the highest concentrations of selenium in serum as further demonstrated by the positive correlation with age. In both pregnant and contraceptive-using women, copper was greatly increased, confirming the influence of estrogen status and/or oral contraceptive intake on increased serum copper levels. However, in postmenopausal women, HRT did not significantly affect serum copper levels. Selenium, copper, and zinc status were not different between normolipidemic and hyperlipidemic subjects for the same gender and age range, but selenium levels tended to increase with hyperlipidemia when considering the whole group of subjects. With respect to the lifestyle, higher serum zinc levels were found in tobacco-consuming men. Albumin serum levels were similar for all considered subgroups, except for the pregnant women, for whom a decrease in this parameter was observed. The present study allowed one to obtain reference values for this healthy group of population, which will serve for a comparative study with groups having pathological conditions, such as cardiovascular disease.     

Effects of Obesity,Glucocorticoid, and Oral Contraceptive Therapy on Plasma Glucose and Blood Pyruvate Levels

The plasma glucose and blood pyruvate levels were determined after oral glucose tolerance test in six groups of women: non-obese and obese controls and in non-obese and obese women receiving glucocorticoid or oral contraceptive therapy. The mean fasting plasma glucose level was similar in all groups, but glucose tolerance was impaired in the obese controls, non-obese women on oral contraceptives or being treated with glucocorticoids, and appreciably impaired in the obese oral contraceptive and glucocorticoid groups compared with mean levels in non-obese subjects of the same groups. Obesity was associated with abnormally raised blood pyruvate levels in response to a glucose tolerance test in all groups. Striking similarities were observed between the responses of the plasma glucose and blood pyruvate levels to glucose tolerance tests in the obese control and non-obese oral contraceptive and non-obese glucocorticoid-treated groups. It is suggested that these abnormalities result from a common mechanism—namely, glucocorticoid excess.     

The differences between aromatizable and non-aromatizable androgens in relation to body composition and metabolic syndrome risk factors in men

The relationships between the parameters of metabolic syndrome and non-aromatizable metabolites of testosterone have been discussed in literature. Some papers describe these metabolites as one of the possible causes of male-type obesity. On the contrary, other studies show a protective influence of dihydrotestosterone on visceral obesity. The aim of this study to analyse the relationship between anthropometric parameters, lipid spectrum, glycemia and the level of endogenous testosterone and dihydrotestosterone, and to compare the effects of these androgens. Our population-based study involved 232 healthy men ranging from 20 to 78 years with BMI 18 to 39 kg/m(2). Serum testosterone, dihydrotestosterone and sex hormone binding globulin SHBG levels, lipid spectrum, glucose metabolism parameters were measured and the oral glucose tolerance test was carried out in all subjects. Their anthropometric parameters (weight, height, waist, hips, waist-to-hip ratio, 14 skin folds) and body composition parameters were determined and calculated by the Antropo program. Multiple regression analysis showed a correlation between hormonal levels, esp. of testosterone and dihydrotestosterone, and the anthropometric data, lipid spectrum and parameters of glucose regulation. Low testosterone and/or dihydrotestosterone was correlated to a higher body-mass index, fat content, waist diameter, total-, HDL-, LDL-cholesterol and triglycerides, fasting glucose, insulin resistance and lower muscle and bone mass. In addition, statistical analysis using multivariate regression with reduction in dimensionality did not discover any striking difference between aromatizable and non-aromatizable androgens in their association to lipid and glucose metabolism parameters in healthy, normosthenic men. In conclusion, the association of endogenous testosterone and dihydrotestosterone to anthropometric data, lipid spectrum and insulin sensitivity are of the same quality; however, the effect of the circulating levels of dihydrotestosterone is quantitatively smaller.     

Skeletal effects of oral oestrogen compared with subcutaneous oestrogen and testosterone in postmenopausal women.

STUDY OBJECTIVE--To compare oral and implanted oestrogens for their effects in preventing postmenopausal osteoporosis. DESIGN--Non-randomised cohort study of postmenopausal women treated with oral or depot oestrogens and postmenopausal controls. SETTING--Gynaecological endocrine clinic in tertiary referral centre. PATIENTS--Oral treatment group of 37 postmenopausal women (mean age 57.5 years, median 8.75 years from last menstrual period), compared with 41 women given oestrogen implants (mean age 56.2 years, median 9.5 years from last menstrual period) and 36 controls (mean age 51.8 years, median 2.0 years from last menstrual period). Weight was not significantly different among the groups. INTERVENTIONS--Oral treatment group was given continuous treatment with cyclic oestrogen and progesterone preparations (Prempak C or Cycloprogynova) for a median of 8.0 years. Implant group was given subcutaneous implants of oestradiol 50 mg combined with testosterone 100 mg, on average six monthly for a median of 8.5 years. Controls were not treated. END POINT--Significant increase in bone density. MEASUREMENTS AND MAIN RESULTS--Bone density measured by dual beam photon absorptiometry was 1.02 (SD 0.13) g hydroxyapatite/cm2 in implant group versus 0.89 (0.11) in oral group (p less than 0.01) and 0.87 (0.14) in controls (p less than 0.01). Serum oestradiol concentration in implant group was (median) 725 pmol/l versus 170 pmol/l in oral group (p less than 0.01) and 99 pmol/l in controls (p less than 0.01). Serum follicular stimulating hormone was median 1 IU/l (range 1-11) in implant group (equivalent to premenopausal values) versus 43 (4-94) IU/l in oral group (p less than 0.01) and 72 (28-99) IU/l in controls (p less than 0.01). CONCLUSIONS--Subcutaneous oestrogen is more effective than oral oestrogen in preventing osteoporosis, probably owing to the more physiological (premenopausal) serum oestradiol concentrations achieved. It also avoids problems of compliance that occur with oral treatment.     

Vitamin A,Pregnancy, and Oral Contraceptives

It has been shown that women receiving oral contraceptives have increased levels of serum vitamin A. High vitamin A levels may constitute a teratogenic hazard and it has been suggested that women who conceive soon after discontinuing oral contraceptive therapy may be especially at risk to this hazard.We have confirmed a significant increase in vitamin A levels in women taking oral contraceptives. During early pregnancy there is no significant difference in vitamin A levels between women who have recently been taking oral contraceptives and those who have not. We have been unable to show that either taking oral contraceptives shortly before pregnancy or a high vitamin A level during the first trimester of pregnancy, comparable to that of a woman taking oral contraceptives, has any detrimental effect on the outcome of pregnancy. It seems unlikely that women who conceive soon after discontinuing oral contraception run any teratogenic risk from increased vitamin A levels.     

Oral contraceptives: effect of folate and vitamin B12 metabolism.

Women who use oral contraceptives have impaired folate metabolism as shown by slightly but significantly lower levels of folate in the serum and the erythrocytes and an increased urinary excretion of formiminoglutamic acid. The vitamin B12 level in their serum is also significantly lower than that of control groups. However, there is no evidence of tissue depletion of vitamin B12 associated with the use of oral contraceptives. The causes and clinical significance of the impairment of folate and vitamin B12 metabolism in these women is discussed in this review of the literature. Clinicians are advised to ensure that women who shop taking "the pill" because they wish to conceive have adequate folate stores before becoming pregnant.     

Effect of oral contraceptives on sebum excretion rate.

Oral contraceptives containing a high dose of oestrogen reduce the sebum excretion rate (SER) and improve acne vulgaris, but more progestogenic preparations may exacerbate acne. The effect on the SER of several oral contraceptives with varying progestogenic potencies was studied in 81 women. The predominantly progestogenic pills (Eugynon 30, Gynovlar) produced no significant change in SER, whereas the rate in women taking a more oestrogenic pill (Minovlar) was significantly reduced compared with the rate in controls. Progestogens therefore do not exacerbate acne by inducing seborrhoea, but in the doses we studied they nullified the inhibitory effect of oestrogens on the sebaceous glands. Acne-prone women who require an oral contraceptive should be given a predominantly oestrogenic preparation.     

Contraception use and associations with intimate partner violence among women in Bangladesh

This study examines the association between contraception use and intimate partner violence (IPV) among women of reproductive age in Bangladesh. The observational study of 10,996 women used the chi-squared test and logistic regressions to assess the associations. Almost 80% of all respondents had used contraceptives at some point in their lives. About half of the respondents (48%) were victims of physical violence, while 11% experienced sexual abuse from their husbands. Urban residents, higher educated women and women aged 20-44 were more likely to use contraceptives than their peers in rural areas, those with lower education and those in their late forties (45-49 years). Women exposed to physical violence were almost two times (OR 1.93, CI 1.55-2.41) more likely to use contraceptives compared with their non-abused peers. Sexual abuse had no significant association with contraceptive use. Physical violence is a predictor for higher levels of contraceptive use among women in Bangladesh. The findings emphasize the importance of screening for IPV at health care centres. The differences in urban and rural contraceptive use and IPV exposure identified by the study have policy implications for service delivery and planning.     

Role of dihydrotestosterone in the control of sexual behaviour on castrated male sheep

Wethers (at least 2 1/2 years after castration) were implanted with testosterone propionate (TP), oestradiol dipropionate (ODP), dihydrotestosterone, or a combination of dihydrotestosterone and ODP Silastic capsules. Active immunization against both oestradiol and oestrone or oestradiol only was used to negate effects of oestrogens produced by aromatization of TP. On exposure to oestrous ewes, immunization of wethers implanted with TP significantly (P less than 0.01) reduced all components of mating behaviour (except sniffing and Flehmen) to levels seen in untreated controls. The results support the conclusion that dihydrotestosterone potentiates the action of oestrogens, particularly as regards Flehmen, and has no action on its own within the central nervous system, while oestrogens do not restore mating activity to the same level as that following treatment with testosterone.     

Oral Contraceptives,Depression, and Libido

Depression, headaches, and libido were rated in 272 women before starting a contraceptive method and at intervals during the first year of use—54 were fitted with an intrauterine device (I.U.D.) and 218 used one of three oral contraceptives. Side effects caused 25% of the oral contraceptive group and 13% of the I.U.D. group to stop the method. Depression, headaches, and loss of libido were the most common reasons for stopping oral contraceptives and breakthrough bleeding was the most common reason for stopping the I.U.D.The group of women who stopped or changed their oral contraceptives during the survey were compared with the group who remained on the same oral contraceptive throughout. The former had higher mean depression and neuroticism scores at the first clinic visit and contained more women with a history of premenstrual weepiness, depression during pregnancy, outpatient psychiatric treatment, and treatment with antidepressants. Changes in the depression, headache, and libido ratings throughout the survey are presented.     

Oral contraceptives and fatal subarachnoid haemorrhage.

A case-control study was conducted of the deaths from subarachnoid haemorrhage (SAH) in women aged 15-44 in England and Wales in 1976. There was a small excess of oral contraceptive use by the women who died from SAH compared with their generally healthy practice-matched controls; this was not, however, statistically significant. Out of 134 women who died from SAH, 34 had a history of hypertension compared with only six of their controls. Renal disease and pre-eclamptic toxaemia were more commonly associated with hypertension in the dead women than in controls. No change in the annual mortality from SAH has been observed in the past 20 years such as might have been expected if the risks were high. Although current or past use of oral contraceptives may have increased the blood pressure and risk of SAH in a few women, the most important factor in determining this risk was hypertension. SAH should thus probably not be regarded as serious cause for concern in healthy non-hypertensive women using oral contraceptives.     

Urinary Tract Dilatation and Oral Contraceptives

Six nulliparous patients were found to have overdistensiblity of the upper urinary tract on excretion urography. Of these five had had persistent or recurrent urinary infections. All had been or were receiving oral contraceptives. Though overdistensibility associated with contraceptive therapy is unlikely to be life-threatening, it may be associated with increased patient morbidity.