Estrone exposure interacts with temperature to alter predator evasion performance and systemic mRNA abundances.

Environmental estrogens from anthropogenic activities are ubiquitous in aquatic ecosystems. Ambient temperature in these systems also fluctuates in daily, seasonal, and long-term rhythms. While both factors have been studied extensively, their interaction on aquatic life is critical to understand. The objective of this study was, therefore, to examine how behavior and gene expression are impacted by estrogenic exposure across a range of environmental temperatures. Larval fathead minnows (Pimephales promelas) were exposed to estrone (E1) at two concentrations (nominal 625 and 1250 ng/L) or to an ethanol solvent control, at one of four temperatures (15, 18, 21 and 24 °C) from fertilization to 21 days post-hatch. Exposed larvae were assessed for alterations in predator evasion performance and mRNA abundances of two genes for calcium channel receptors found in muscles - dihydropyridine receptor (dhpr) and ryanodine receptor 1, and the gonadal genes anti-Müllerian hormone, cytochrome P450 gonadal aromatase (cyp19a), doublesex and mab-3 related transcription factor 1 (dmrt1) and estrogen receptor 1 (esr1). Larval escape angle, escape latency, as well as systemic esr1 and cyp19a mRNA abundances were altered by an interaction between E1 concentration and temperature. E1-exposed larval exhibited reduced escape performance across all tested temperatures, whereas decreased systemic dhpr mRNA abundance was observed only at 18 °C. E1-exposure reduced systemic mRNA abundances of amh, cyp19a, dhpr, and ryr1, while temperature significantly reduced systemic cyp19a and dhpr mRNA abundances. E1-exposure and temperature significant enhanced systemic mRNA abundances of esr1 and cyp19a, respectively. These complex results illustrate the importance of considering how abiotic factors may moderate the effects of contaminant exposure during the sensitive larval developmental stage, as temperature modulates effects of estrogenic exposure on animal performance and mRNA abundances.

Temperature modulates estrone degradation and biological effects of exposure in fathead minnows.

Environmental pollutants, including estrogens, are widespread in aquatic environments frequently as a result of treated wastewater effluent discharged. Exposure to estrogens has been correlated with disruption of the normal physiological and reproductive function in aquatic organisms, which could impair the sustainability of exposed populations. However, assessing the effects of estrogen exposure on individuals is complicated by the fact that rates of chemical uptake and environmental degradation are temperature dependent. Because annual temperature regimes often coincide with critical periods of biological activity, temperature-dependent changes in estrogen degradation efficacy during wastewater treatment could modulate biological effects. We examined the interactions between ambient water temperature and degradation of estrone (E1) during wastewater treatment. In addition, we exposed mature fathead minnows (Pimephales promelas) to three environmentally relevant concentrations of E1 at four different water temperatures (15°C, 18°C, 21°C, and 24°C) to reflect natural seasonal variation. E1 degradation occurred with and without the support of robust nitrification at all temperatures; however, the onset of E1 degradation was delayed at cooler water temperatures. In addition, we observed significant interactive effects between temperature and E1 exposure. Female morphometric endpoints were more susceptible to temperature-modulating effects while physiological endpoints were more strongly affected in males. Collectively, the data demonstrate that natural seasonal fluctuations in temperature are sufficient to affect E1 degradation during wastewater treatment and induce sex-dependent physiological and anatomical changes in exposed fish.

Distribución de esteroides estrogénicos en plantas de tratamiento de aguas residuales municipales / Distribution of estrogenic steroids in municipal wastewater treatment plants

En la actualidad, existe una preocupación creciente por la presencia de estrógenos en el medio acuático, donde pueden ser introducidos a partir de aguas residuales después de su eliminación incompleta en las plantas de tratamiento. Las aguas residuales sistemáticamente reciben estrógenos naturales y sintéticos, y por lo tanto una comprensión más profunda de la suerte de ellos en el medio ambiente es necesaria. Se evaluaron los niveles de estrógenos en los efluentes de las Plantas de Tratamiento de Aguas Residuales (PTARs) Penha e Ilha do Governador, ambos de tipo convencional de flujo continuo de lodo activado con aireación prolongada. Fue utilizado como el parámetro de determinación de algunos compuestos de interés como estrógenos naturales [estrona (E1), 17β-estradiol (E2), estriol (E3)) y sintéticos (17α-etinilestradiol (EE2)]. Las muestras individuales se recogieron posteriormente al tratamiento de cada PTAR y después de los procedimientos de laboratorio se realizó la determinación de estrógenos basado en la extracción en fase sólida (SPE) y la cromatografía líquida de alta resolución con detector de arreglo de diodos (HPLC-DAD). Las concentraciones fueron de: 0,7 a 5,2 μg/l y de 0,5 a 5,6 de E1; 0,9 a 7,7 y 1,2 a 9,2 μg/l para E2; 2,01 a 6,09 y 1,07 a 4,08 μg/l para EE2 en PTAR Penha y PTAR Ilha do Governador, respectivamente. La capacidad de eliminación de estrógenos fue eficaz, pero denota que la eliminación sistemática de la población es en la actualidad alta. Se recomienda instalar mecanismos para mitigar el consumo exagerado de estas sustancias o implementar una eliminación completa más eficaz.

Currently, there is a growing concern over the presence of estrogens in the aquatic environment, where they can be introduced from wastewater after their incomplete elimination in the treatment plants. Wastewater systematically receives natural and synthetic estrogens, and thus a deeper understanding of the fate of them in the environment is extremely necessary. It was evaluated estrogen levels in the effluent from the Sludge Wastewater Treatment Plants (SWTPs) Penha and Ilha do Governador, both of type conventional continuous-flow activated sludge with extended aeration. The determination of some target compounds as natural estrogens was used as the evaluation parameter [estrone (E1), 17β-estradiol (E2), estriol (E3) and synthetic (17α-ethinylestradiol (EE2)]. Individual samples were collected posterior treatment of each SWTP, and after laboratory procedures, the determination of estrogens was performed by a method based on solid phase extraction (SPE) and high performance liquid chromatography-diode array detector (HPLC-DAD). Concentrations ranged from 0.7 to 5.2 μg/l and from 0.5 to 5.6 for E1; 0.9 to 7.7 and 1.2 to 9.2 μg/l for E2; 2.1 to 6.9 and 1.7 to 4.8 μg/l for EE2 at SWTPs Penha and Ilha do Governador, respectively. The removal capacity of estrogens despite its effectiveness denotes that the systematic elimination by the population is high nowadays and urging mechanisms to mitigate the exaggerated consumption or to implement most effective complete removal.

Impact of environmental estrogens on nucleotide excision repair gene expression in embryonic zebrafish.

Estrogens and estrogen mimics are aquatic contaminants that can elicit a variety of deleterious effects in exposed fauna. One of the most potent xenoestrogens found in the aquatic environment is 17α-ethinylestradiol (EE(2)), the pharmaceutically derived semi-synthetic hormone found in oral contraceptives and hormone replacement therapies. Exposure to 100 ng/L EE(2) has previously been shown to profoundly decrease functional hepatic nucleotide excision repair (NER) processes in adult zebrafish in correlation with dramatic decreases in the abundance of hepatic XPC and XPA transcripts; however, its effects on these processes in embryos are currently unknown. Because developing organisms are known to have increased sensitivities to endocrine disrupting compounds such as EE(2), the goal of this study was to examine the impacts of estrogen exposure on mRNA expression of these two key NER genes in zebrafish embryos during the first 4 days of development. Embryos were exposed from 0 h post fertilization (hpf) to waterborne EE(2), its major metabolite, estrone (E(1)), or combinations of the two compounds and sampled at 12, 24, 48, 72 and 96 hpf. Increased abundance of vitellogenin-1 (VTG1) mRNA, a bioindicator of estrogen exposure, was evident as early as 24 hpf in embryos that were co-exposed to EE(2) and E(1) and this effect was sustained throughout 96 hpf. Embryos exposed to EE(2) alone exhibited elevated VTG1 beginning at 72 hpf. In contrast to observations from adult zebrafish exposed to EE(2), embryos did not show any change in mRNA abundance of the excision repair gene, XPC, during the first 4 days of development. However, co-exposure to EE(2) and E(1) elicited an increase in XPA mRNA abundance at 48 and 72 hpf, which was the opposite response as that observed in exposed adults where hepatic XPA mRNA abundance decreased after EE(2) exposure. These differences between embryos and adults suggest that alteration of NER gene transcription by EE(2) is operating under different stimuli during development.

Beware of registries for their biases.

Patient registries are very popular. On the other hand, scientific data collections in registries are commonly observational and retrospective and, in many instances, are prone to biases. Same thing is true of administrative data bases. The selection of the control group(s) is probably the Achilles heel of scientific data collection in observational studies, and there are historical examples of how a properly chosen control group can help or its absence deceive us. Somewhat more recently recognized biases are the wandering comparisons of risk, confounding by disease severity, channeling bias, depletion of the susceptible, and the immortal time bias. The last bias can especially be deceiving and give us false hopes of new remedies. A particularly important selection bias we have come across is what we call the "mortality bias." This is where the mortality in a mother population lessens the mortality in the registry that stems from this mother population simply because deaths in the former cannot be represented in the latter.

Continuous estrone treatment impairs spatial memory and does not impact number of basal forebrain cholinergic neurons in the surgically menopausal middle-aged rat.

CEE (conjugated equine estrogens) is the most widely prescribed estrogen-only menopausal hormone therapy in the United States, and is comprised of over 50% estrone (E1) sulfate. Following CEE administration, E1 is the principal circulating estrogen. However, the cognitive and neurobiological effects of E1 in a middle-aged rodent model have not yet been evaluated. We assessed cognitive effects of continuous E1 treatment in middle-aged surgically menopausal rats using a maze battery. We also quantified number of choline acetyltransferase-immunoreactive (ChAT-IR) neurons in distinct basal forebrain regions known in earlier studies in to be impacted by the most potent naturally-circulating estrogen in rodents and women, 17ß-estradiol (17ß-E2), as well as CEE. On the spatial working memory delayed-match-to-sample water maze, the highest E1 dose impaired memory performance during acquisition and after delay challenge. E1 did not impact ChAT-IR neuron number in the medial septum (MS) or horizontal/vertical diagonal bands. In a comparison study, 17ß-E2 increased MS ChAT-IR neuron number. Findings indicate that E1 negatively impacts spatial working memory and memory retention, and does not increase ChAT-IR neuron number in basal forebrain, as does 17ß-E2. Thus, data from prior studies suggest that 17ß-E2 and CEE can enhance cognition and increase number of ChAT-IR basal forebrain neurons, while here we show that E1 does not induce these effects. Findings from preclinical basic science studies can inform the design of specific combinations of estrogens that could be beneficial to the brain and cognition. Accumulating data suggest that E1 is not likely to be among these key beneficial estrogens.

Development of an LC-MS/MS method to quantify sex hormones in bovine milk and influence of pregnancy in their levels.

Hormones work in harmony in the body, and this status must be maintained to avoid metabolic disequilibrium and the subsequent illness. Besides, it has been reported that exogenous steroids (presence in the environment and food products) influence the development of several important illnesses in humans. Endogenous steroid hormones in food of animal origin are unavoidable as they occur naturally in these products. The presence of hormones in food has been connected with several human health problems. Bovine milk contains considerable quantities of hormones and it is of particular concern. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method, based on hydroxylamine derivatisation, has been developed and validated for the quantification of six sex hormones in milk [pregnenolone (P5), progesterone (P4), estrone (E1), testosterone (T), androstenedione (A) and dehydroepiandrosterone (DHEA)]. This method has been applied to real raw milk samples and the existence of differences between milk from pregnant and non-pregnant cows has been statistically confirmed. Basing on a revision of existing published data, it could be concluded that maximum daily intakes for hormones are not reached through milk ingestion. Although dairy products are an important source of hormones, other products of animal origin must be considered as well for intake calculations.

Comparing predicted against measured steroid estrogen concentrations and the associated risk in two United Kingdom river catchments.

Predicted concentrations of estrone, 17ß-estradiol, and 17α-ethinylestradiol generated from a geographical information systems-based model (LF2000-WQX) have previously been used to assess the risk of causing intersex in male fish in the rivers of England and Wales, United Kingdom. Few measured data of sufficient quality and spatial extent have been available to verify this risk assessment. New measured data have been collected from sewage treatment plant effluents and the receiving waters upstream and downstream of these discharges from the Erewash River and the Avon River systems in England. The model results for these rivers were in good agreement with the measured values in terms of estradiol equivalents. Critically, the risk assessment based on the measured data gave a risk assessment nearly identical to that derived from the modeled results. For individual estrogens, 17α-ethinylestradiol was modeled best and estrone worst. Poor simulations reflected poor estimates of the effluent concentrations, which were more variable from day to day and between works of nominally similar type than is assumed in the model. In support of this, model results for the Erewash River, calculated using observed effluent concentrations, were in excellent agreement with the measured data. The model has proved to be adequate in predicting overall estrogenic potency, and therefore risk, along these rivers; however, improvements are possible, particularly in predicting STP removal efficiency and therefore effluent concentrations.

Effects of unintentional exposure of children to compounded transdermal sex hormone therapy.

Gynecomastia and rapid growth progressed in twin brothers and pubic hair in one, over a period of 2 years. A combination of contra- and isosexual development was induced by transdermal exposure to compounded estradiol, estrone, and testosterone creams applied to their mother's body as part of a hormone replacement regimen.

The effect of estrone on thrombin generation may explain the different thrombotic risk between oral and transdermal hormone replacement therapy.

BACKGROUND: The metabolism of estrogen contained within hormone replacement therapy (HRT) is influenced by the route of administration, and this may affect the risk of venous thromboembolism. Thrombin generation, a global coagulation assay, is a marker of hypercoagulability and is of potential use in determining the thrombotic risk associated with particular HRT administration routes. OBJECTIVES: To determine whether any effect of oral and transdermal HRT on thrombin generation is related to the plasma estrogen profile. METHODS: We investigated the effects of oral, transdermal and no HRT (controls) in 52, 39 and 52 postmenopausal women, respectively, on thrombin generation, standard markers of thrombophilia, estradiol level and estrone level. RESULTS: All parameters of thrombin generation were altered in women using oral HRT as compared with controls (P<0.001 for all comparisons). No such differences were found in women using transdermal HRT. Estrone levels correlated with peak thrombin generation (R=0.451, P<0.001) in women using oral HRT, but there was no correlation in women using the transdermal route. CONCLUSIONS: Thrombin generation is significantly increased in women who use HRT administered by the oral route. This is probably mediated by the hepatic first-pass metabolism of estrone, the main metabolite of oral estradiol, which is avoided by the transdermal route. The effect of estrone on thrombin generation may provide the explanation for the higher thrombotic risk seen in women using oral rather than transdermal HRT.

Molecular clustering of endometrial carcinoma based on estrogen-induced gene expression.

Identification of biomarkers potentially provides prognostic information that can help guide clinical decision-making. Given the relationship between estrogen exposure and endometrial cancer, especially low grade endometrioid carcinoma, we hypothesized that high expression of genes induced by estrogen would identify low risk endometrioid endometrial cancers. cDNA microarray and qRT-PCR verification were used to identify six genes that are highly induced by estrogen in the endometrium. These estrogen-induced biomarkers were quantified in 72 endometrial carcinomas by qRT-PCR. Unsupervised cluster analysis was performed, with expression data correlated to tumor characteristics. Time to recurrence by cluster was analyzed using the Kaplan-Meier method. A receiver operating characteristic (ROC) curve was generated to determine the potential clinical utility of the biomarker panel to predict prognosis. Expression of all genes was higher in endometrioid carcinomas compared to non-endometrioid carcinomas. Unsupervised cluster analysis revealed two distinct groups based on gene expression. The high expression cluster was characterized by lower age, higher BMI, and low grade endometrioid histology. The low expression cluster had a recurrence rate 4.35 times higher than the high expression cluster. ROC analysis allowed for the prediction of stage and grade with a false negative rate of 4.8% based on level of gene expression in endometrioid tumors. We have therefore identified a panel of estrogen-induced genes that have potential utility in predicting endometrial cancer stage and recurrence risk. This proof-of-concept study demonstrates that biomarker analysis may play a role in clinical decision making for the therapy of women with endometrial cancer.

[Piperazinyl estrone, a new estrogen derivant, prevents bone loss in aged rats].

The purpose of this study was to determine the effects of piperazinyl estrone, a new estrogen derivative, on bone turnover, bone mass and uterine weight in female aged rats. Thirty-two Sprague-Dawley female rats at the age of 22 months were treated with vehicle or with piperazinyl estrone (P-E) at 0.5, 1 and 2 mg/kg/day, subcutaneous injection for 1 month. At the time of death, the uterine weight was measured and bone histomorphometric analysis of proximal tibial metaphyses (PTM) was performed in undecalcified sections. Compared with control, bone mass was increased in P-E groups. Dynamic data showed that bone resorption were decreased, but bone formation was not declined and bone mass was increased significantly in P-E (1 mg/kg day) group. There was no significant change in uterine weight. The findings of this study show that piperazinyl estrone at dosage of 1 mg/kg/d is most efficacious in preventing the bone losses in aged rats and has no side effect on uterus.

Synthesis and reactivity of the catechol metabolites from the equine estrogen, 8,9-dehydroestrone.

The risk factors for women developing breast and endometrial cancers are all associated with a lifetime of estrogen exposure. Estrogen replacement therapy in particular has been correlated with an increased cancer risk. Previously, we showed that the equine estrogens equilin and equilenin, which are major components of the widely prescribed estrogen replacement formulation Premarin, are metabolized to highly cytotoxic quinoids which caused oxidative stress and alkylation of DNA in vitro [Bolton, J. L., Pisha, E., Zhang, F., and Qiu, S. Chem. Res. Toxicol. 1998, 11, 1113-1127]. In this study, we have synthesized 8,9-dehydroestrone (a third equine estrogen component of Premarin) and its potential catechol metabolites, 4-hydroxy-8,9-dehydroestrone and 2-hydroxy-8,9-dehydroestrone. Both 2-hydroxy-8,9-dehydroestrone and 4-hydroxy-8,9-dehydroestrone were oxidized by tyrosinase or rat liver microsomes to o-quinones which reacted with GSH to give one mono-GSH conjugate and two di-GSH conjugates. Like endogenous estrogens, 8,9-dehydroestrone was primarily converted by rat liver microsomes to the 2-hydroxylated rather than the 4-hydroxylated o-quinone GSH conjugates; the ratio of 2-hydroxy-8,9-dehydroestrone versus 4-hydroxy-8,9-dehydroestrone was 6:1. Also in contrast to experiments with equilin, 4-hydroxyequilenin was not observed in microsomal incubations with 8,9-dehydroestrone or its catechols. The behavior of 2-hydroxy-8,9-dehydroestrone was found to be more complex than 4-hydroxy-8,9-dehydroestrone as GSH conjugates resulting from 2-hydroxy-8,9-dehydroestrone were detected even without oxidative enzyme catalysis. Under physiological conditions, 2-hydroxy-8,9-dehydroestrone isomerized to 2-hydroxyequilenin to form the very stable 2-hydroxyequilenin catechol; however, 4-hydroxy-8,9-dehydroestrone was found to be stable under similar conditions. Finally, preliminary studies conducted with the human breast tumor S-30 cell lines demonstrated that the catechol metabolites of 8,9-dehydroestrone were much less toxic than 4-hydroxyequilenin (20-40-fold). These results suggest that the catechol metabolites of 8,9-dehydroestrone may have the ability to cause cytotoxicity in vivo primarily through formation of o-quinones; however, most of the adverse effects of Premarin estrogens are likely due to formation of 4-hydroxyequilenin o-quinone from equilin and equilenin.

Piperazine oestrone sulphate and interrupted norethisterone in postmenopausal women: effects on bone mass, lipoprotein metabolism, climacteric symptoms, and adverse effects.

OBJECTIVE: To compare the effects of two doses of piperazine oestrone sulphate combined with interrupted norethisterone, with that of oestradiol continuously combined with norethisterone acetate, and with placebo, in postmenopausal women. DESIGN: A prospective randomised trial. PARTICIPANTS: Two hundred postmenopausal women. SETTING: Monocentre study with expertise in osteoporosis. METHODS: The participants were randomly assigned to two years of treatment with alternating three-day cycles of 1.5 mg of piperazine oestrone sulphate plus 0.7 mg of norethisterone (highEP), or alternating three-day cycles of 0.75 mg of piperaine oestrone sulphate plus 0.35 mg of norethisterone (lowEP), or 2 mg of 17beta-oestradiol continuously combined with 1 mg of norethisterone acetate (E2+NETA), or placebo. MAIN OUTCOME MEASURES: Change in bone mineral density, lipoprotein metabolism, climacteric symptoms, and adverse effects. RESULTS: One hundred and twenty-one women completed the study. Spinal bone mineral density was increased about 9% over two years by E2+NETA, about 6% by highEP, 4% by lowEP, but remained unchanged in the placebo group. The same pattern was seen in the hip and forearm. All hormone regimens decreased markers of bone turnover and alleviated climacteric symptoms. Serum lipoproteins decreased by about 10% in all hormone groups. CONCLUSIONS: All hormone regimens studied prevented bone loss completely and lowered serum lipids.

Successful treatment of non-segmental vitiligo: systemic therapy with sex hormone-thyroid powder mixture.

We previously reported a patient with generalized vitiligo improved by oral administration of the drug for menopausal syndrome (sex hormone-thyroid powder mixture). In this study, we reevaluated the efficiency of this drug for vitiligo, and examined its pharmacological action in melanogenesis.

Low-dose esterified estrogens (0.3 mg/day): long-term and short-term effects on menopausal symptoms and quality of life in postmenopausal women.

OBJECTIVE: To study the impact of low-dose unopposed esterified estrogens on menopausal symptoms and quality of life. METHODS: In a long-term, 2-year, randomized, double-blind, placebo-controlled study, 204 postmenopausal women were treated with esterified estrogens 0.3 mg daily or placebo. Menopausal symptoms were assessed with a modified Kupperman index at baseline, 3, 6 and thereafter every 6 months. In a second 12-week, open-label, short-term pilot study, 25 postmenopausal women with moderate to severe vasomotor symptoms were treated with esterified estrogens 0.3 mg daily for 12 weeks. Vasomotor symptoms and quality of life were assessed using the Greene scale and Quality of Life Menopause Scale (QUALMS). RESULTS: In the long-term study, significant (p < 0.05) reductions in total symptom scores were observed at each time point with esterified estrogens compared with placebo. Somatic symptom scores (hot flushes, night sweats, vaginal dryness) decreased significantly (p < 0.01) in patients treated with esterified estrogens 0.3 mg compared to baseline and placebo. In the short-term, open-label pilot study, the incidence of vasomotor symptoms was significantly (p < 0.01) reduced with esterified estrogens 0.3 mg from week 4 until the study end. Significant (p < 0.05) improvements versus baseline were seen in the somatic and vasomotor/sleep domains and in the total quality-of-life score. CONCLUSIONS: Esterified estrogens 0.3 mg given daily provide adequate menopausal symptom relief and improved quality of life in postmenopausal women.

[Dermatitis caused by estrogens]. / Dermatite aux oestrogènes.

BACKGROUND: We report a case of sensitization to estrogen. CASE REPORT: A 40-year-old woman consulted for skin disorders which followed a cyclic pattern. At each menses, the patient developed pruritus and erythematous papulovesicular lesions over the members and trunk. Estraderm patch contact dermatitis was evident. Prick and patch tests with alcoholic solutions of estrone alone were positive. Serum tests were positive for anti-ethinyl-estradiol antibodies and anti-progesterone antibodies. DISCUSSION: Autoimmune dermatitis can be caused by sensitization to endogenous or exogenous sex hormones. Clinical manifestations and histological findings are variable and non-specific. The cyclic nature of the manifestations is however quite suggestive. Positive prick and patch tests performed with alcohol solutions of the hormones may give the diagnosis and serum tests may be positive for specific anti-steroid antibodies. These complementary explorations are however difficult to perform and interpret and definitive diagnosis is based on an association of clinical findings, skin tests, laboratory tests and the clinical course. In case of progesterone sensitization, the treatment of choice is estrogen inhibition of ovulation. For estrogen sensitization, anti-estrogen treatment appears to be more effective. Finally, bilateral ovariectomy may be required in difficult cases.

Bleeding pattern and endometrial changes during continuous combined hormone replacement therapy. The Ogen/Provera Study Group.

OBJECTIVE: To establish the optimum oral daily dose of micronized medroxyprogesterone acetate, given in combination with a fixed oral dose of estrone (E1) sulfate as hormone replacement therapy, that provides endometrial protection and induces cessation of vaginal bleeding. METHODS: This multicenter, randomized, double-blind study was conducted for 2 years. Five hundred sixty-eight postmenopausal women were randomized to take E1 sulfate 1.25 mg daily and one of three doses of medroxyprogesterone acetate (2.5, 5, or 10 mg) daily. Any vaginal bleeding was recorded by patients in a daily diary, and endometrial biopsies were performed at entry into the study and at 3, 12, and 24 months. RESULTS: Forty-two percent of all women reported some bleeding at month 3 of therapy. However, by month 6, 76.5, 80.1, and 80.9% of women were amenorrheic in the 2.5-, 5-, and 10-mg medroxyprogesterone acetate groups, respectively. Over time, the percentage of women with no bleeding increased in each group, and by 24 months 91.5, 89.9, and 94.3% were amenorrheic in the 2.5- and 10-mg medroxyprogesterone acetate groups, respectively. Approximately 10% of women continue to have some bleeding, regardless of the dose of medroxyprogesterone acetate. There were no statistically significant differences in the number of women with bleeding at any time point between the three groups. There were no cases of endometrial hyperplasia reported in the study population over the 2 years. CONCLUSION: All three studied doses of medroxyprogesterone acetate, given in combination with 1.25 mg of E1 sulfate, provide adequate endometrial protection and render approximately 80% of women amenorrheic by 6 months of therapy.

Menopausal symptom control and side-effects on continuous estrone sulfate and three doses of medroxyprogesterone acetate. Ogen/Provera Study Group.

OBJECTIVES: To establish the optimum oral daily dose of micronized medroxyprogesterone acetate (MPA), given in combination with 1.25 mg of estrone sulfate for menopausal symptom control. METHODS: This multicenter, randomized, double-blind study was conducted on 568 postmenopausal women who were randomized to take estrone sulfate 1.25 mg daily with 2.5, 5.0 or 10 mg of MPA daily for 2 years. The number of vasomotor symptoms and the severity of mood swings, lethargy, vaginal dryness and loss of libido as well as side-effects were recorded in a diary. Blood pressure and weight were recorded at each 3-month visit. RESULTS: Vasomotor symptoms were reported by approximately 80% of subjects at month 1, 23% at month 3 but only 9% by month 24. Mood swings, lethargy and vaginal dryness improved rapidly in the initial 3 months of therapy. Decrease in libido had a slower response to therapy in all three treatment groups. Breast tenderness was the commonest side-effect with 22% of subjects complaining of this in the first 3 months of therapy, dropping to 13% by 6 months. Headache, depression, nausea, bloating and irritability showed a similar pattern of decline. There was no significant difference in the rate of decrease in menopausal symptoms or reported side-effects between the three treatment groups. There was a small but significant (p < 0.001) decrease in systolic and diastolic blood pressure over the study period. CONCLUSIONS: All three treatment regimens provide adequate symptom control. Side-effects decreased markedly after the first 3 months, with no significant difference between the treatment groups.