Influence of augmented Hageman factor (Factor XII) titers on the cryoactivation of plasma prorenin in women using oral contraceptive agents.

Prolonged cold storage of plasma may induce the conversion of plasma prorenin (inactive renin) to renin. This phenomenon is exaggerated in oral contraceptive (OC) users; the titer of Hageman factor (HF, Factor XII) in OC users is higher than in nonusers. The present study relates these observations. The increment in plasma renin activity (PRA) during cold storage, as measured by generation of angiotensin I, correlated strongly with the initial plasma titer of HF. Increasing the HF titer of nonusers to that observed in OC users by addition of purified HF increased cold-induced PRA at least twofold, while reducing the plasma HF titer of OC users correspondingly decreased cold-induced PRA. Thus, in OC users, the enhanced conversion of plasma prorenin to renin during cold storage reflects the elevated plasma titer of HF.

PIP: Prolonged cold storage of plasma may induce the conversion of plasma prorenin (inactive renin) to renin. This phenomenon is exaggerated in oral contraceptive (OC) users; the titer of Hageman factor (HF, Factor 12) in OC users is higher than in nonusers. The present study relates these observations. The increment in plasma renin activity (PRA) during cold storage, as measured by generation of angiotensin I, correlated strongly with the initial plasma titer of HF. Increasing the HF titer of nonusers to that observed in OC users by the addition of purified HF increased cold-induced PRA at least 2-fold, while reducing the plasma HF titer of OC users correspondingly decreased cold-induced PRA. Thus, in OC users, the enhanced conversion of plasma prorenin to renin during cold storage reflects the elevated plasma titer of HF.

Clinical behavior of a cohort of adult women with facial acne treated with combined oral contraceptive: ethinylestradiol 20 µg/dienogest 2 mg.

Acne vulgaris is the most common skin disease. It affects the young adult female population and generates great impact on physical and mental health. One of the treatments with good results for affected women is combined oral contraceptive pills (COCPs). The aim of this study was to determine the clinical effect of facial acne management with ethinylestradiol 20 µg/dienogest 2 mg in a cohort of Colombian adult women. A cohort of 120 female university students was followed for 12 months. These participants were enrolled in the Sexual and Reproductive Health Program of the Santiago de Cali University. This cohort admitted women between 18 and 30 years old who had chosen to start birth control with ethinylestradiol 20 µg/dienogest 2 mg COCPs, did not have contraindi cations to the use of COCPs, and had been diagnosed with acne. Monthly monitoring of facial acne lesion count was performed. Relative changes in facial lesion count were identified. At the end of follow-up, the percentage of reduction of lesions was 94% and 23% of women had a 100% reduction in acne lesions. In conclusion, the continued use of the ethinylestradiol 20 µg/dienogest 2 mg COCPs reduced inflammatory and non-inflammatory acne lesions in reproductive-age women between 18 and 30 years of age with no severe acne.

Oral contraceptive use and fibrocystic breast disease with special reference to its histopathology.

The relationship between use of oral contraceptives and fibrocystic breast disease was examined in a hospital-based case-control study undertaken in New Haven, Connecticut, from 1977 to 1979. Particular emphasis was placed on the extent of epithelial atypia and other histopathologic characteristics found in the biopsy specimens from the cases. Women who had ever used oral contraceptives were at a somewhat decreased risk for fibrocystic disease as a whole. Cases with high atypia and controls had similar patterns of oral contraceptive use, whereas cases with low and intermediate atypia had less oral contraceptive use than controls. Cases with intermediate atypia reported the lowest oral contraceptive use. Subjects with biopsy specimens exhibiting gross cysts, microscopic cysts, or papillomatosis were about 50% less likely to have used oral contraceptives than controls.

Mammary nodules in dogs during four years' treatment with megestrol acetate or chlormadinone acetate.

PIP: A 7 year study of megestrol and chlormadinone in female dogs is in progress. This report characterized histopathologically 60 mammary nodules during the first 4 years of the study. 100 purebred female beagles, 6-12 months of age, were randomly assigned to 5 equal groups. One group was used as a control. Oral doses were .01, .10, and .25 mg/kg/day of megestrol acetate in coconut oil in capsules and of chlormadinone acetate .25 mg/kg/day in lactose tablets. These doses were 1, 10, and 25 times the projected dose of megestrol for humans and about 25 times the human dose of chlormadinone. After 2 years 4 dogs from each group were necropsied. One high-dose megestrol-treated and 1 chlormadinone-treated dog had benign mixed mammary tumors. Palpable nodules were first observed at 16 months in the chlormadinone-treated dogs, at 18 months in dogs given the high dose megestrol and at 27 months in the dogs treated with middle-dose megestrol. Transitory nodules were found in 4 control dogs after 21 months and in low dose megestrol-treated dogs at 26 months. Of 38 grossly detected nodules evaluated microscopically from the megestrol-treated dogs 27 were nodular hyperplasia, 5 were benign mixed mammary tumors, 3 were ductal dialatations, 1 was a lymph node, 1 was fat necrosis and 1 was the umbilicus. Of 22 nodules from the chlormadinone-treated dogs 12 were nodular hyperplasia, 4 benign mixed mammary tumors, 1 chondromucoid degeneration and 1 adenocarcinoma with widespread metastases. 3 nodules were lymph nodes and 1 other had no mammary tissue. Involutions, regression and sclerosis of many areas of nodular hyperplasia were evident at 4 years. Thus of the 60 nodules evaluated during the first 4 years of the study 50 were non-neoplastic and 10 were neoplastic. It is considered that the 1 adenocarcinoma may have been spontaneous and not a treatment-related neoplasm. A precursor stage through nodular hyperplasia apparently did not occur.^ieng

Lack of an elevated risk of malignant melanoma in relation to oral contraceptive use.

Epidemiologic and laboratory data suggest an effect of oral contraceptives (OC) on the risk of malignant melanoma. This relationship was explored in a hospital-based case-control study of 160 women with malignant melanoma and 640 matched controls, all of whom were white and 20-59 years of age. A total of 63 cases (39%) had used OC compared with 270 controls (42%), yielding a relative risk estimate of 0.9 (95% confidence interval: 0.6-1.3). When a number of potential confounding factors were simultaneously controlled, the relative risk estimate was 0.8 (0.5-1.3). For use that lasted 5 or more years the estimate was 0.9 (0.5-1.6). The level of tumor invasion was not related to OC use. The evidence from this study suggests that OC, even when used for 5 or more years, do not increase the risk of malignant melanoma.

Oral contraceptive use and fibrocystic breast disease of different histologic classifications.

The relationship between oral contraceptive (OC) use and occurrence of fibrocystic breast disease (FBD) of different histologic classifications was evaluated with data from a cohort study. Biopsy specimens from 232 women with FBD were classified into different atypia categories. In 96 matched pairs of OC users and nonusers, atypia scores were lower in users than in nonusers. Women without breast diseases (500 OC users and 500 nonusers) were sampled from the original cohort to form a two-stage "anamorphic" study with the 232 cases of FBD. The previously shown inverse association between OC use and FBD occurrence was present and increased with increased length of OC use. However, the "protective effect" of OC use did not vary for different histologic classifications of FBD. The findings from both paired and anamorphic analyses of the data are not consistent with the hypothesis that the use of OC is associated with decreased frequency only of FBD with minimal epithelial atypia.

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

Breast cancer epidemiology.

PIP: The various risk factors for breast cancer have been recognized for many years. A table lists these established breast cancer risk factors together with the approximate magnitude of the increase in risk associated with them. Breast cancer incidence rates increase with age throughout the life span in Western countries, although the rate of increase is greater up to age 50 years than after 50 years. Breast cancer is more common among women in upper rather than lower social classes, among women who never have been married, among women living in urban areas, among women living in the northern US than in the southern US, and among whites than blacks, at least among those over age 50. Women in North American and Northern European countries have the highest risk for breast cancer, women in Southern European and Latin American countries are at intermediate risk, and women in Africa and Asian countries have the lowest risk. Yet, rapid rates of increase in incident rates have been noted in recent years in many Asian, Central European, and some South American countries. The later the age at which a woman has her 1st full-term pregnancy, the higher her risk for breast cancer; the earlier the age at menarche and the later the age at menopause the higher the risk; and among women who have a premenopausal oophorectomy, the earlier the age at which this occurs the lower the risk. Among postmenopausal women, obesity is associated with an increase in risk. Lactation is negatively associated with subsequent breast cancer risk. Some current research is considering potential risk factors that have not been well studied in the past, including alcohol consumption, cigarette smoking, caffeine consumption, exposure to diethylstilbestrol (DES), emotional stress, exposure to electric power, and lack of physical activity. Other areas of current research reviewed here include radiation, mammographic parenchymal patterns, a high-fat diet, use of oral contraceptives (OCs), use of estrogen replacement therapy, and endogenous hormones. Cigarette smoking and caffeine consumption do not appear promising as potential etiologic agents. The studies of the DES-exposed women and of OC users suggest that the timing of exposure may be critical, since the possible effect of both these hormonal agents may be limited to specific time periods of rapid breast development. If such a critical period does not exist in postmenopausal women, then there may be little effect of hormones used at this time. Studies with long-term follow-up and that include long-term users are essential to studies of effects of hormones and other exposures.^ieng

Breast carcinoma etiological factors.

PIP: This review of breast carcinoma etiological factors considers the following: heritage; menses, marital state, and parity; breastfeeding; contraceptives; benign epithelial diseases of the breast; hormonal factors; cancer; iatrogenic factors; immunological factors; viral aspects of human breast cancer; dietary factors; and psychosomatic factors. Breast carcinoma is the most prevalent type of cancer in American women. 8% of American women can expect to be stricken with this disease. The rate in men is 1% that in women. The causes or reasons for a woman being afflicted with this disease remain equivocal. An abundance of evidence exists that the incidence of breast carcinoma varies greatly from 1 population to another throughout the world and that in most populations it is increasing. Due to earlier detection, improved medical care, and possible other factors, the death rate is not increasing as rapidly as the incidence rate. In general, the incidence is greatest in populations with the highest standards of living, such as those of Northwestern Europe and North America. Thus, a woman's heritage is usually a large factor in determining her low risk of developing mammary carcinoma. Heritage includes family, race, country of origin, religion, and any component of lifestyle that is firmly passed on from 1 generation to the next. These factors seem to have a great influence on the incidence of breast cancer, but there is little agreement on which components of heritage are most important and how they operate. Many publications report insignificant or no effect of menarchal age on breast cancer risk. Many reports mention breastfeeding along with other data in breast cancer etiological studies, but its influence on the disease seems to be insignificant. Possibly there is some synergistic influence, but available data is not strong enough to establish the direction. Many reports seem to suggest that oral contraceptives (OCs) increase risk in nulliparous women and may promote the growth of malignant neoplasia, but in parious women, if there is no incipient cancer present, OCs do not appear to increase risk and may even decrease risk. Benign epithelial lesions can be harbingers of breast carcinoma, but from the literature it is not easy to conclude the nature of the most risky lesions nor their relative risks. A familial or personal history of cancer in any location or tissue may increase the risk of breast cancer. Ionizing radiation, oophorectomy, hysterectomy, and hormone therapy are the principal iatrogenic factors that influence the incidence of breast cancer. Radiation is known to be a most powerful carcinogen and is probably responsible for more cancer than any other iatrogenic factor. None of the factors so far studied is of great importance singly, but in a situation where several act together, any one may appear to be significant.^ieng

Implantable controlled release systems.

PIP: A controlled release system utilizes a polymer matrix or pump as a rate-controlling device to deliver the drug in a fixed, predetermined pattern for a desired time period. These systems have the following advantages compared to other methods of administration: 1) plasma drug levels are continuously maintained in a therapeutically desirable range, 2) harmful side effects from systemic administration can be reduced or eliminated by local administration from a controlled release system, 3) drug administration may be improved and facilitated in underpriviledged areas where good medical supervision is not available, 4) administration of drugs that have short in vivo half lives may be greatly facilitated, 5) continuous small amounts of drug may be less painful than several large doses, 6) patient compliance may be improved, and 7) this method is relatively less expensive and less wasteful of the drug. Disadvantages include possible toxicity, need for surgery to implant the system, possible pain, and difficulty in shutting off release if necessary. Two types of diffusion-controlled systems have been developed. The reservoir is a core of drug surrounded with a polymer film. The 2nd type, the matrix, is one in which the drug is uniformly distributed through a polymer. In chemically controlled systems, the rate of drug release is regulated by a chemical reaction with the polymer. In solvent activated systems a swelling or osmotic mechanism is involved. Pharmaceutical applications have been made in ocular disease with the Ocusert, a reservoir system for glaucoma therapy and which is not widely used, and in contraception with 4 systems. These systems are: 1) subdermal implants of nonbiodegradable polymers such as Norplant, 6 capsules of 36 mg levonorgestrel; 2) subdermal implant of biodegradable polymers; 3) steroid releasing IUD; and 4) vaginal rings, which are silicone-coated. Other applications have been made in the areas of dentistry, immunization, anticoagulation, cancer, narcotic antogonists, and insulin delivery. Transdermal delivery involves placing a polymeric system containing a contact adhesive on the skin.^ieng

Androgen antagonists in androgen target tissues.

PIP: Most antiandrogens appear to act by binding to the androgen receptor and competitively inhibiting the binding of testosterone and cihydrotestosterone to the receptor. Focusing on those compounds which appear to inhibit androgen receptor mediated responses, this review discusses the chemistry of those antiandrogens which have been studied to the extent that their mechanism of action is at least partially understood, outlines the mechanism of androgen action as it is currently understood and suggests how antiandrogens might fit in with this mechanism, indicates the major metabolites of several important antiandrogens, and discusses the clinical applications of several antiandrogens. Cyproterone acetate has been studied extensively as a potential male contraceptive. Although it was recognized that 100 mg of cyproterone acetate per day inhibited spermatogenesis, that dose also reduced libido and potency. Following the administration of 10 or 20 mg of cyproterone acetate per day to 15 males for 26 weeks, the following observations were made: the number of motile sperm was reduced; the quality of their motion was impaired; and the ability of the sperm to penetrate cervical mucus was decreased. Sperm density was also suppressed, but neither it nor sperm motility were inhibited to the extent necessary for contraception. Antiandrogens have been demonstrated to be beneficial in treating 5 clinical syndromes or diseases: acne, seborrhea, hirsutism with or without menstrual abnormalities; precocious puberty; benign prostatic hypertrophy; cancer of the prostate; and sexual deviates. Since 3 of these conditions are very common, effective and safe treatment would have a large market. At this time, antiandrogens are widely used in Europe for treatment of seborrhea, acne, and hirsutism and a large Veterans Administration Cooperative Study in the US was approved but has not yet been funded to compare antiandrogens with other treatments for cancer of the prostate. Studies to assess antiandrogen interaction with other hormones or drugs have been limited. Side effects in the female have been best evaluated when cyproterone acetate was administered in combination with ethinyl estradiol. In 46 women followed over 317 cycles, side effects were similar to those reported with estrogen-progestin contraceptives. Administration of 10-20 mg of cyrproterone acetate per day to males caused no significant side effects, but 100 mg or more/day has caused loss of libido, impotence, gynecomastia, tiredness, weakness, decreased efficiency, weight gain, drying and desquamation of skin over the legs, and loss of hair on the trunk and pubic area.^ieng

Normal adults and subjects with hypogonadotropic hypogonadism respond differently to D-Ser(TBU)6-LH-RH-EA10.

Daily administration of 5microgram of D-Ser(TBU)6-LH-RH-EA10 for one week produced a significant increase in the LH response to GnRH in hypogonadotropic hypogonadal subjects and a significant decrease in the response of normal male adults. Basal plasma testosterone concentrations fell in normal controls but were unchanged in the hypogonadal group.

PIP: Daily administration of 5 mcg of the gonadotropin-releasing hormone (GnRH) analogue D-Ser (TBU) 6-LH-RH-EA 10 for 1 week produced a significant increase in the luteinizing hormone (LH) response to GnRH in hypogonadotrophic hypogonadal subjects and a significant decrease in the response of normal male adults. Subjects included 4 men and 2 women with an isolated gonadotropin deficiency and 4 men and 1 woman with hypogonadism secondary to proven craniopharyngioma; 5 adult men of proven fertility served as controls. All sex steroid treatment was discontinued at least 3 months before the study. Basal plasma testosterone concentrations fell in controls but were unchanged in the hypogonadal group 9 days after onset of treatment with the analogue. This difference in responsiveness to LH between normal and hypogonadotrophic hypogonadal subjects has also been recorded following prolonged therapy with natural GnRH. Moreover, this finding implies that the results of Hoe 766 studies in normal subjects cannot be extrapolated. Patients with isolated gonadotropin deficiency showed greater increments in LH secretion than those with craniopharyngiomas. The 3 patients who had not received prior gonadal steroid therapy responded relatively poorly in terms of LH increment, perhaps indicating that the administration of gonadal steroids is associated with increased LH responsiveness and an adult pattern of gonadotropin secretion. The tendency for plasma testosterone levels to rise in the hypogonadotrophic hypogonadal subjects in this study suggests that long-acting GnRH analogues have potential for use in this area.

Homologous radioimmunoassay for human epidermal growth factor (urogastrone).

Epidermal growth factor (EGF), a polypeptide hormone originally discovered in the mouse submaxillary gland, stimulates growth in a variety of tissues in several species. This hormone has recently been identified in human urine. A homologous RIA for human EGF (RIA-hEGF) has been developed. In general, levels were similar to those recently reported using a heterologous RIA system. Twenty-four-hour urinary excretion of RIA-hEGF by normal adult males and females was 63.0 +/- 3.0 and 52.0 +/- 3.5 (mean +/- SE) micrograms/total vol, or 29.7 +/- 1.1 and 39.8 +/- 1.7 micrograms/g creatinine, respectively. Excretion by females taking oral contraceptives was significantly greater (60.1 +/- 2.7 micrograms/g creatinine; P less than 0.01) than that by females who were not. Recent evidence suggests the probable identity of hEGF and beta-urogastrone, a potent inhibitor of gastric acid secretion. Adult males with active peptic ulcer disease appeared to have lower urinary RIA-hEGF excretion (22.9 +/- 2.6 micrograms/g creatinine) than normal men, but this was not significant (P greater than 0.05). Several of those with very low values had histories of alcohol abuse. Excretion by patients with Cushing's syndrome was normal. Patients with psoriasis or recovering from major burns excreted both abnormally high and abnormally low levels of RIA-hEGF, with no obvious correlation to their clinical condition. There was no apparent diurnal or postprandial variation in urinary RIA-hEGF excretion by normal subjects. An excellent linear correlation was observed between RIA-hEGF and creatinine concentrations in each urine sample for each subject, suggesting that RIA-hEGF concentration in a random urine sample provides a valid index of 24-h RIA-hEGF excretion.

Combined effects of RU486 and tamoxifen on the growth and cell cycle phases of the MCF-7 cell line.

The antiproliferative effect of RU486 and its effect combined with tamoxifen on the growth and cell cycle kinetics parameters on the MCF-7 human carcinoma cells were investigated. When MCF-7 cells in the exponential growth phase were treated with RU486 (10(2) nmol/L), a time-dependent cell growth inhibition was observed which was significant 5 days after the beginning of treatment. This inhibition was accompanied by a time- and dose-dependent decrease in the percentage of S and G2-M phase cells and a concomitant increase in the percentage of cells in the G0/G1 phase of the cell cycle. With tamoxifen (10(5) pmol/L), growth inhibition was obtained after 4 days of treatment of cells, and the blockage of the cell cycle occurred in the G0/G1 phase. In the case of simultaneous treatment of MCF-7 cultures with RU486 (10(2) nmol/L) and tamoxifen (10(5) pmol/L), we observed a synergistic inhibitory effect on the proliferative rate for short treatment (less than or equal to 3 days), whereas RU486 or tamoxifen alone had no effect. For the intermediate treatment (4 days), the combined effect of RU486 and tamoxifen was not significant compared to the effect of tamoxifen alone. For the long treatment (greater than 4 days), there were no differences between the number of cells in the treated cultures under different experimental conditions, but all were inhibited compared to those in control cell cultures. This simultaneous treatment of cells does not induce any change in the distribution of cells in the different cell cycle phases compared to tamoxifen percentages. These results suggest that RU486 is a cell cycle phase-specific growth inhibitory agent, and a combination of antiestrogen/antiprogestin should be considered as a possible improvement in breast cancer endocrine therapy.

PIP: The antiproliferative effect of the antiprogestational agent RU-486 (100 nmol/L), alone and its additive effect in combination with the antiestrogenic tamoxifen (100,000 pmol/L) on the growth and cell cycle phases of the human breast cancer cell line MCF7, which is ER and PR positive, using image cytometry was investigated. On days 2, 3, 4, 5, 7, and 9, cells were harvested from control and RU-486-, tamoxifen, and RU-486- and tamoxifen-treated cultures and counted with an hemocytometer. Cell number increased in the RU-486-treated culture for the first 4 days as well as in the control culture. RU-486 treatment of cells for 5 days produced a significant time-dependent cell growth inhibition (P 0.01) compared with that in the control cells. With tamoxifen (100,000 pmol/L), growth inhibition was obtained after 4 days of treatment of cells, and the blockage of the cell cycle occurred in the G0.G1 phase. For the short treatment a significant synergistic growth inhibition (P 0.05 and P 0.01 on days 2 and 3, respectively) compared to RU-486 or tamoxifen alone, which had no effect compared to control cells. In the case of simultaneous treatment of MCF7 cultures with RU-486 (100 nmol/L) and tamoxifen (10000 pmol/L), a synergistic inhibitory effect of the proliferative rate was not produce an effect. During intermediate treatment (4 days), the combined effect of RU-486 and tamoxifen was not significant compared to the effect of tamoxifen alone. On the other hand, tamoxifen inhibited growth compared to that in control cells (P 0.05). These findings suggest that RU-486 is a cell cycle phase-specific growth inhibitory agent, and a combination of antiestrogen and antiprogestin agents may be of possible use in breast cancer endocrine therapy.

Impairment of prednisolone disposition in women taking oral contraceptives or conjugated estrogens.

Companion studies were designed to determine the effects of oral contraceptives and conjugated estrogens on the pharmacokinetics of prednisolone. Twenty-four normal women entered the studies, including six young women taking oral contraceptives and six age-matched control women, and six postmenopausal women receiving conjugated estrogens and six age-matched postmenopausal women. All received 0.53 mg/kg prednisolone phosphate, iv. Significant decreases (P less than 0.05) in the clearance and volume of distribution and significant increases in the half-life were found for both total and unbound prednisolone in the women taking oral contraceptives compared to values in the young control women. A significant decrease in the unbound clearance and increases in the total and unbound half-lives of prednisolone were found in the women receiving conjugated estrogens compared to values in the postmenopausal control women. Total clearance and volume of distribution were unchanged by conjugated estrogen therapy. Administration of prednisolone to women receiving estrogen-containing oral contraceptives or conjugated estrogens results in exposure of these women to increased concentrations of unbound prednisolone for increased periods of time. Increases in the pharmacological and toxic effects of prednisolone might be expected in these women.

The secretion of human chorionic gonadotropin-like substance in women employing contraceptive measures.

To study the secretion pattern of a hCG-like substance (hCG') in normal women and those employing contraceptive measures, we assayed daily urine concentrates by RIAs using an anti-hCG beta-COOH-peptide serum and an anti-hCG serum to monitor hCG' and human LH (hLH), respectively. In eight cycles of four normal women, urinary hCG' was within the normal range (less than 100 ng/day), except that four samples in two women had marginal elevations of hCG' during the menstrual or premenstrual period, but not at the time of the midcycle hLH surge. Among seven cycles from three intrauterine device (IUD) users, six showed midcycle hLH surges, and in five, hCG' was prominent (0.17-3.6 micrograms/day) in the late luteal phase. In eight cycles of women ingesting steroid contraceptives on a conventional schedule, hCG' was found during the premenstrual and menstrual intervals. The pattern of hCG' secretion was episodic, and hCG' levels were in the range of 0.3-1.8 microgram/day. It was notable that hCG' was excreted at the highest levels during intervals when ingestion of steroids was withheld. These findings suggest that contraceptive steroid hormones may modulate pituitary hCG' secretion. From gel high pressure liquid chromatographic studies, the immunoactive hCG' detected in urinary extracts from normally cycling women and IUD users had a molecular size larger than hCG beta and hLH, but was slightly smaller than hCG (lot CR119). The hCG' identified in these eluents is unlikely to be a subunit of hCG. On the other hand, the immunoactive hCG' in the urinary extracts from oral contraceptive users was mainly the hCG beta-subunit-like substance. Both hCG alpha and hCG beta immunoactive substances were heterogeneous in size in contrast to those found in normally cycling women and IUD users. In summary, the hCG' excretion pattern was different in menstrual and postmenopausal women, and hCG' identified in these subjects was heterogeneous.

PIP: To study the secretion pattern of an hCG-like substance (hCG') in normal women and those employing contraception, the authors assayed daily urine concentrates by radioimmunoassays using an anti-hCGbeta-COOH-peptide serum and an anti-hCG serum to monitor hCG' and human luteinizing hormone (hLH), respectively. In 8 cycles of 4 normal women, urinary hCG' was within the normal range (100 ng/day), except that 4 samples in 2 women had marginal elevations of hCG' during the menstrual or premenstrual period, but not at the time of the midcycle hLH surge. Among 7 cycles from 3 IUD users, 6 showed midcycle hLH surges, and in 5, hCG' was prominent (0.17-3.6 mcg/day) in the late luteal phase. In 8 cycles of women ingesting steroid contraceptives on a conventional schedule, hCG' was found during the premenstrual and menstrual intervals. The pattern of hCG' secretion was episodic and hCG' levels were in the range of 0.3-1.8 mcg/day. It was notable that hCG' was excreted at the highest levels during intervals when steroid ingestion was withheld. These findings suggest that contraceptive steroids may modulate pituitary hCG' secretion. From gel high pressure liquid chromatographic studies, the immunoactive hCG' detected in urinary extracts from normally cycling women and IUD users had a molecular size larger than hCGbeta and hLH, but was slightly smaller than hCG (lot CR119). The hCG' identified in these eluents is unlikely to be a subunit of hCG. On the other hand, the immunoactive hCG' in the urinary extracts from OC users was mainly the hCG beta-subunit-like substance. Both hCGalpha and hCGbeta immunoactive substances were heterogeneous in size in contrast to those found in normally cycling women and IUD users. In summary, the hCG' excretion pattern was different in menstrual and postmenopausal women, and hCG' identified in these subjects was heterogeneous.

Association of melasma with thyroid autoimmunity and other thyroidal abnormalities and their relationship to the origin of the melasma.

Melasma is localized hyperpigmentation over the forehead, upper lips, cheeks, and chin. In this study, evidence suggesting an association between autoimmune thyroid disorders and melasma and the relationship of thyroid disorders to the origin of melasma is presented. A total of 108 nonpregnant women, aged 20-56 yr, were divided into 2 groups for the purpose of this study: 1) melasma, 84 patients; 2) control group, 24 patients from the Dermatology Clinic matched for age and sex. Microsomal thyroid autoantibodies (MCHA) were sought in all subjects. TRH-TSH tests were performed in patients with melasma and in those women with goiter and/or positive MCHA tests from the control group. Studies were completed with serum T4, T3, and antithyroglobulin antibody (TGHA) measurements in all patients with thyroid abnormalities. In patients with melasma, the frequency of thyroid disorders (58.3%) was 4 times greater than in the control group. The MCHA-negative patients had 1) simple goiter (13.1%), 2) Plummer's disease (2.4%), and 3) TSH hyperresponse to TRH in nongoitrous patients (10.7%). Patients with positive MCHA tests (32.1%) were divided into 2 subgroups. One comprised those women with an apparently normal thyroid gland and thyroid function (n = 7), while the other included all patients with goiter and/or subclinical hypothyroidism (n = 20). Regarding the origin of the melasma, it was found that 70% of women who developed melasma during pregnancy or while using oral contraceptives had thyroid abnormalities compared to 39.4% of patients with idiopathic melasma. Subjects from the control group had a 12.5% incidence of thyroid abnormalities, and only 8.3% had positive MCHA. Estrogen, progesterone, or both could be the triggering factor in the development of melasma in women who have a particular predisposition toward both melasma and thyroid autoimmunity. Patients with idiopathic melasma had a lower frequency of thyroid abnormalities, suggesting that there may be different genetic patterns linked to autoimmune thyroid disease. We conclude that there is a true association between thyroid autoimmunity and melasma, mostly in women whose melasma develops during pregnancy or after ingestion of oral contraceptive drugs.

The circadian pattern of melatonin and its positive relationship with progesterone in women.

To investigate the possible role of melatonin in the regulation of the human menstrual cycle, the circadian pattern of melatonin was determined in the follicular and luteal phases of 10 normal women. Four-hourly sampling was used to derive a melatonin index which described the total exposure to melatonin for 24 h. This sampling procedure adequately represented the circadian melatonin output and demonstrated that pulses of melatonin secretion, inconsistent with a measured half-life of 47 min, did not exist. A significant increase (P less than 0.001) in the melatonin index was found in the luteal phase compared to that in the follicular phase. To investigate the influence of exogenous progestins on the melatonin pattern, repeated 24-h profiles were measured in 8 women taking the 3-phase contraceptive pill. There was a significant increase (P less than 0.01) in the melatonin index associated with an increase in the dose of progestin. These results are consistent with a positive relationship between melatonin and progesterone and suggest that changes in the circadian pattern of melatonin secretion, rising during the luteal phase with a fall before ovulation, may act as a modulator of cyclicity.

PIP: 10 normal women participated in a study designed to investigate the possible role of melatonin in the regulation of the human menstrual cycle. The circadian pattern of melatonin was determined in the follicular and luteal phases of the study subjects who had conventional life styles and normal sleep/wake rhythms. Also investigated was the influence of exogenous steroids on the circadian pattern of melatonin, using subjects who were taking a 3-phase oral contraceptive (OC). Initially, 2 studies were performed to determine the frequency of sampling necessary to define the circadian profile of melatonin and to determine the relevance of that sampling frequency to the half-life of melatonin. 3 routes of administration were employed: 2 mg melatonin as a 0.04% solution in corn oil containing 2% ethanol, taken orally as a suspension in 50 ml milk; 2 mg melatonin in an 8% solution of ethanol in a nasal spray; and 2 mg melatonin in 250 mg ascorbic acid, taken orally in a gelatin capsule. A program of 4-hourly sampling was adopted. The 10 healthy women ranged in age from 20-40 years and had normal menstrual cycles. A circadian pattern of serum melatonin concentrations was observed in all study subjects. Figure 1 shows the 24-hour profiles of melatonin in 1 woman studied in the follicular and luteal phases of her menstrual cycle. The concentrations of melatonin in samples taken at intervals of 15 minutes for a 24-hour period indicated that episodic release did not take place. The increase in the melatonin index from the follicular to the luteal phase was highly significant for the group as a whole. The investigation of other aspects of the circadian pattern of melatonin showed a significant increase in the luteal phase compared to the follicular phase in peak serum melatonin concentrations, estimated from data at 4-hourly intervals. The mean duration of elevation was 11.6 +- 0.8 h in the luteal phase and 10.7 +- 0.7 h in the follicular phase. All subjects had a single peak concentration, at either 2400 or 0400 h. A change in the melatonin index was found in the 8 women using the 3-phase OC pill when sampled at 2 times during the cycle. A significant increase in the melatonin index was associated with an increase in the dose of progestin, either from no progestin intake to a low-dose or from a low-dose to a higher dose.

The role of estrogen in the modulation of pituitary sensitivity of LRF (luteinizing hormone-releasing factor) in men.

The role of estradiol in modulating pituitary gonadotropin release in the human male was studied by evaluating the effects of clomiphene on the pituitary gonadotropin response to synthetic LRF (150 mug) in 6 eugonadal men. Administration of clomiphene (100 mg single daily dose time 5 days) induced a significant elevation of the basal levels of LH, FSH, estradiol and testosterone. However, the pituitary release of LH and FSH in response to LRF was markedly diminished by the clomiphene treatment. This finding suggests that in men, as in women, endogenous estradiol provides feedback regulation of gonadotropin output by the pituitary.

PIP: The effects of clomiphene on the pituitary response to LRF in eugonadal men were studied to investigate the role of estradiol in the feedback modulation of gonadotropin secretion in men. 150 mcg LRF was given to 6 healthy men (20-28 years) and luteinizing hormone (LH) and follicle stimulating hormone (FSH) evaluated. A 5-day course of clomiphene (100 mg daily) was then given and the LRF test repeated. Serum FSH, LH, estradiol, and testosterone (T) were determined. Clomiphene caused a significant elevation in the basal levels of LH, FSH, Estradiol, and T. Pituitary release of LH and FSH in response to LRF was markedly reduced by clomiphene treatment indicating that estradiol provides feedback regulation of gonadotropin output by the pituitary.

Fourteen-day versus twenty-one-day regimens of intermittent intranasal luteinizing hormone-releasing hormone agonist combined with an oral progestogen as antiovulatory contraceptive approach.

This study was designed to determine the effect of discontinuous administration of a LHRH agonist on pituitary-ovarian function in normal women. The LHRH agonist buserelin (200 micrograms/12 h or 400 micrograms/24 h) was given intranasally for four consecutive cycles for 14 or 21 days in 26 normally cycling women. Five milligrams of medroxyprogesterone acetate were given orally twice daily from days 15-21. There was a 7-day pause between each medication cycle. Blood samples were drawn every other day for RIA of LH, FSH, estradiol (E2), and progesterone (P). Serum FSH increased for only a few days at the beginning of each cycle, whereas sustained elevation of serum LH occurred during LHRH agonist administration. Serum E2 increased rapidly and remained elevated during the administration of buserelin. Serum P remained in the follicular phase range or increased briefly after the initiation of buserelin occasionally in the 14-day regimens. After discontinuation of buserelin, E2 fell rapidly, and uterine withdrawal bleeding occurred. During the pause, FSH increased progressively. The patterns of gonadotropin response to buserelin were similar in the four cycles. Based on measurement of the areas of the response curves, serum LH and E2 levels were higher during the administration of 200 micrograms/12 h compared to 400 micrograms/24 h buserelin. However, down-regulation of the pituitary-ovarian axis, as evaluated by the acute gonadotropin response to buserelin on day 14, was more pronounced with 200 micrograms/12 h than with 400 micrograms/24 h. Breakthrough bleeding occurred in the 14-day schedules, whereas withdrawal bleeding occurred during the pause in the 21-day schedules. The immediate cycles following buserelin administration were normal ovulatory cycles. Intermittent LHRH agonist administration for 21 days avoided constant down-regulation of the pituitary-ovarian axis and allowed regular uterine bleeding. Combined with an appropriate P complement, it could be a useful contraceptive approach.