Evaluation of analytical methodology for the detection of hormones and their attenuation during aquifer recharge and recovery cycles.

The hormones listed in the screening survey list 2 of the Unregulated Contaminant Monitoring Rule 3 (estrone, 17-ß-estradiol, 17-α-ethynylestradiol, 16-α-hydroxyestradiol (estriol), equilin, testosterone and 4-androstene-3,17-dione) were analyzed by liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). Two analytical methods were compared: EPA method 539 and the isotope dilution method. EPA method 539 was successfully utilized in river and drinking water matrices with fortified recoveries of 98.9 to 108.5%. Samples from the Hillsborough River reflected levels below the method detection limit (MDL) for the majority of the analytes, except estrone (E1), which was detected at very low concentrations (<0.5 to 1 ng L(-1)) in the majority of samples. No hormones were detected in drinking water samples. The isotope dilution method was used to analyze reclaimed and aquifer storage and recovery (ASR) water samples as a result of strong matrix/solid phase extraction (SPE) losses observed in these more complex matrices. Most of the compounds were not detected or found at relatively low concentrations in the ASR samples. Attenuation of 50 to 99.1% was observed as a result of the ASR recharge/recovery cycles for most of the hormones, except for estriol (E3). Relatively stable concentrations of E3 were found, with only 10% attenuation at one of the sites and no measureable attenuation at another location. These results have substantiated that while EPA method 539 works well for most environmental samples, the isotope dilution method is more robust when dealing with complex matrices such as reclaimed and ASR samples.

Contraceptive options and their associated estrogenic environmental loads: relationships and trade-offs.

This work explores the relationships between a user's choice of a given contraceptive option and the load of steroidal estrogens that can be associated with that choice. Family planning data for the USA served as a basis for the analysis. The results showed that collectively the use of contraception in the USA conservatively averts the release of approximately 4.8 tonnes of estradiol equivalents to the environment. 35% of the estrogenic load released over the course of all experienced pregnancies events and 34% the estrogenic load represented by all resultant legacies are a result of contraception failure and the non-use of contraception. A scenario analysis conducted to explore the impacts of discontinuing the use of ethinylestradiol-based oral contraceptives revealed that this would not only result in a 1.7-fold increase in the estrogenic loading of the users, but the users would also be expected to experience undesired family planning outcomes at a rate that is 3.3 times higher. Additional scenario analyses in which ethinylestradiol-based oral contraceptive users were modeled as having switched entirely to the use of male condoms, diaphragms or copper IUDs suggested that whether a higher or lower estrogenic load can be associated with the switching population depends on the typical failure rates of the options adopted following discontinuation. And, finally, it was estimated that, in the USA, at most 13% of the annual estrogenic load can be averted by fully meeting the contraceptive needs of the population. Therefore, while the issue of estrogen impacts on the environment cannot be addressed solely by meeting the population's contraceptive needs, a significant fraction of the estrogenic mass released to environment can be averted by improving the level with which their contraceptive needs are met.

Degradation of the potential rodent contraceptive quinestrol and elimination of its estrogenic activity in soil and water.

Quinestrol has shown potential for use in the fertility control of the plateau pika population of the Qinghai-Tibet Plateau. However, the environmental safety and fate of this compound are still obscure. Our study investigated degradation of quinestrol in a local soil and aquatic system for the first time. The results indicate that the degradation of quinestrol follows first-order kinetics in both soil and water, with a dissipation half-life of approximately 16.0 days in local soil. Microbial activity heavily influenced the degradation of quinestrol, with 41.2% removal in non-sterile soil comparing to 4.8% removal in sterile soil after incubation of 10 days. The half-lives in neutral water (pH 7.4) were 0.75 h when exposed to UV light (λ = 365 nm) whereas they became 2.63 h when exposed to visible light (λ > 400 nm). Acidic conditions facilitated quinestrol degradation in water with shorter half-lives of 1.04 and 1.47 h in pH 4.0 and pH 5.0 solutions, respectively. Moreover, both the soil and water treatment systems efficiently eliminated the estrogenic activity of quinestrol. Results presented herein clarify the complete degradation of quinestrol in a relatively short time. The ecological and environmental safety of this compound needs further investigation.

Simultaneous determination of ethinyl estradiol and drospirenone in oral contraceptive by high performance liquid chromatography

A simple, rapid, economical and reliable high performance liquid chromatographic method has been developed and successfully applied in simultaneous determination of ethinyl estradiol and drospirenone in coated tablets. The HPLC method was performed on a LiChroCART® 100RP column (125x4 mm i.d., 5 µm) with acetonitrile:water 50:50 (v/v) as mobile phase, pumped at a flow rate of 1.0 mL.min-1. The fluorescence detection for ethinyl estradiol was made at λex= 280 nm and λem= 310 nm and a UV detection for drospirenone was made at 200 nm. The elution time for ethinyl estradiol and drospirenone were 4.0 and 5.7 min, respectively. The method was validated in accordance to USP 34 guidelines. The proposed HPLC method presented advantages over reported methods and is suitable for quality control assays of ethinyl estradiol and drospirenone in coated tablets.

Um método simples, rápido, econômico e confiável foi desenvolvido empregando a cromatografia líquida de alta eficiência para a determinação simultânea de etinilestradiol e drospirenona em comprimidos revestidos. O método foi realizado utilizando coluna LiChroCART® 100RP (125 x 4 mm d.i., 5 µm), a fase móvel constituída de acetonitrila:água, 50:50 (v/v) com vazão de 1,0 mL.min-1. A detecção foi realizada empregando fluorescência em λex= 280 nm e λem= 310 nm para o etinilestradiol e na região de UV em 200 nm para a drospirenona. O etinilestradiol e a drospirenona tiveram tempo de retenção de 4,0 e 5,7 min, respectivamente. O método foi validado de acordo com as diretrizes da USP 34. O método proposto apresentou vantagens sobre os relatados na literatura e pode ser considerado adequado para o controle de qualidade do etinilestradiol e da drospirenona em comprimidos revestidos.

Tissue-specific uptake and bioconcentration of the oral contraceptive norethindrone in two freshwater fishes.

The environmental presence of the oral contraceptive norethindrone (NET) has been reported and shown to have reproductive effects in fish at environmentally realistic exposure levels. The current study examined bioconcentration potential of NET in fathead minnow (Pimephales promelas) and channel catfish (Ictalurus punctatus). Fathead minnows were exposed to 50 µg/l NET for 28 days and allowed to depurate in clean water for 14 days. In a minimized 14-day test design, catfish were exposed to 100 µg/l NET for 7 days followed by 7-day depuration. In the fathead test, tissues (muscle, liver, and kidneys) were sampled during the uptake (days 1, 3, 7, 14, and 28) and depuration (days 35 and 42) phases. In the catfish test, muscle, liver, gill, brain, and plasma were collected during the uptake (days 1, 3, and 7) and depuration (day 14) stages. NET tissue levels were determined by gas chromatography-mass spectrometry (GC-MS). Accumulation of NET in tissues was greatest in liver followed by plasma, gill, brain, and muscle. Tissue-specific bioconcentration factors (BCFs) ranged from 2.6 to 40.8. Although NET has been reported to elicit reproductive effects in fish, the present study indicated a low potential to bioconcentrate in aquatic biota.

Métodos anticonceptivos: entonces y ahora / Contraceptive methods: then and now

Comparar los métodos anticonceptivos utilizados en la década de 1990 con los utilizados en la década de 2000. Unidad de Ginecología Reproducción y Salud Integral, Valencia, Edo. Carabobo. La muestra la integraron 1 000 pacientes, 500 en 1997-1998 (grupo A) y 500 en 2007-2008 (grupo B). Revisamos las historias médicas de las pacientes que practicaban anticoncepción. Predominaron los anticonceptivos orales en ambos grupos con 29,6 por ciento (grupo A) y 31,4 por ciento (grupo B). Los de mayor uso fueron levonorgestrel+etinilestradiol (25,9 por ciento) y drospirenona+etinilestradiol (24,7 por ciento) respectivamente. En el grupo A los anticonceptivos inyectables fueron mayormente acetato de medroxiprogesterona (66,7 por ciento), y en el grupo B sólo noretindrona+valerianato de estradiol (100 por ciento). El uso de implantes no superó el 1 por ciento en ambos grupos. En el grupo B 8,4 por ciento usaban métodos naturales en contraposición a 23,4 por ciento utilizados en el A. El grupo A usó preservativos en 17,8 por ciento comparado con el grupo B 24,4 por ciento. La esterilización en la cesárea es el método irreversible más frecuente (grupo A 56,9 por ciento y grupo B 67,5 por ciento). No hubo esterilización por laparoscopia en el grupo A, pero sí en el grupo B (10,3 por ciento). En solo una década, se aprecian diferencias significativas en los métodos anticonceptivos que utilizan nuestras mujeres. Los métodos anticonceptivos más utilizados siguen siendo los de administración oral aunque su composición y dosis han sufrido variaciones. El empleo de preservativos ha aumentado. El uso de los métodos naturales ha disminuido. Aparecieron nuevos métodos como los de emergencia, parches y anillos vaginales. La laparoscopia se ha popularizado como método de esterilización

To compare contraceptive methods utilized during 1990 decade with those utilized during decade 2000. Unidad de Ginecologia Reproduccion y Salud Integral, Valencia, Edo. Carabobo. One thousand Unidad de Ginecología Reproducción y Salud Integral patients charts were reviewed, 500 belonged to decade of 1990 (group A) and 500 to decade of 2000 (group B). All patients were utilizing some contraceptive method. Oral contraceptives predominated in both groups (29.6 percent in group A and 31.4 percent in group B). The most utilized contraceptives were levonorgestrel+ethynilestradiol (25.9 percent) and drospirenone+ethynilestradiol (24.7 percent) respectively. In group A injectable contraceptive mainly utilized were medroxyprogesterone acetate (66.7 percent) and only norethindrone+estradiol valerianate (100 percent) in group B. Use of implants didn’t surpass 1 percent in both groups. Group A utilized natural methods in 23.4 percent contrarily to group B that only utilized them in 8.4 percent. Condoms were utilized in 17.8 percent in group A compared with 24.4 percent in group B. Surgical sterilization during cesarean section continues to be the most frequent irreversible method (56 percent in group A and 67.5 percent in group B). There were no laparoscopic sterilizations in group A in opposition to group B in which there were 10.3 percent. In only one decade significative differences in contraceptive methods utilized in our women are observed. The most frequent contraceptive utilized continues to be the oral contraceptives even if its composition and dose have suffered variations. Use of condoms has increased, use of natural methods has decreased, new methods have appeared like emergency contraceptives, patches and vaginal rings. Laparoscopic sterilization has become a popular method of sterilization

Are oral contraceptives a significant contributor to the estrogenicity of drinking water?

Recent observed feminization of aquatic animals has raised concerns about estrogenic compounds in water supplies and the potential for these chemicals to reach drinking water. Public perception frequently attributes this feminization to oral contraceptives (OCs) in wastewater and raises concerns that exposure to OCs in drinking water may contribute to the recent rise in human reproductive problems. This paper reviews the literature regarding various sources of estrogens, in surface, source and drinking water, with an emphasis on the active molecule that comes from OCs. It includes discussion of the various agricultural, industrial, and municipal sources and outlines the contributions of estrogenic chemicals to the estrogenicity of waterways and estimates that the risk of exposure to synthetic estrogens in drinking water on human health is negligible. This paper also provides recommendations for strategies to better understand all the potential sources of estrogenic compounds in the environment and possibilities to reduce the levels of estrogenic chemicals in the water supply.

Initial photocatalytic degradation intermediates/pathways of 17alpha-ethynylestradiol: effect of pH and methanol.

This study investigated the photocatalytic degradation of the synthetic oral contraceptive 17alpha-ethynylestradiol (EE2). Particular attention was paid on the effects of pH and the co-solvent methanol on the degradation intermediates of EE2. Twelve intermediates were identified, and several intermediates reported herein have not been found in previous studies. The degradation efficiency of EE2 and the number of identified intermediates decreased evidently at pH 3 and in the presence of methanol at pH 7. Three photocatalytic degradation pathways were proposed: The transformation of the phenolic moiety (pathway I) is the primary initial reaction pathway; the initial photocatalytic degradation in the aliphatic carbon linked to the aromatic ring (pathway II) only took place at pH 7; the isomerization of EE2 (pathway III) could occur only in the presence of methanol at pH 7. Results from this study underscore the importance of photocatalytic degradation on the removal of estrogenic activity mainly expressed by the phenol moiety of EE2.

[Combined estrogenic activity of soybean extract used in a dietary supplement and ethinyl estradiol].

We examined the combined estrogenic activity of soybean extract used in a dietary supplement and ethinyl estradiol (EE) contained in an oral contraceptive. Olive oil (control), soybean extract (0.0036 or 0.36 g/kg corresponding to doses of total isoflavone of 0.83 or 83 mg/kg respectively), EE (1 or 10 microg/kg), and soybean extract+EE were administered to ovariectomized CD-1 mice by oral gavage for 4 consecutive days. Soybean extract (0.0036 or 0.36 g/kg) and EE (1 microg/kg) did not increase the relative uterine weight. The relative uterine weight of the soybean extract (0.0036 or 0.36 g/kg)+EE (10 microg/kg) group was significantly higher than that of the control. The relative uterine weight of the soybean extract (0.36 g/kg)+EE (10 microg/kg) group was also significantly higher than that of the EE (10 microg/kg) group. Soybean extract showed estrogenic activity for human estrogen receptor (hER)-alpha and -beta. Coadministration of EE with soybean extract increased the estrogenic activity for hER-alpha and -beta.

Serum folate and Vitamin B12 levels in women using modern oral contraceptives (OC) containing 20 microg ethinyl estradiol.

OBJECTIVE: The effects of modern oral contraceptives (OC) on serum concentrations of folate and cobalamin are controversial. STUDY DESIGN: Case-control study on the cobalamin and folate status of 71 healthy female nulligravidae using "low dose" OC for >or=3 months and 170 controls. Factors interfering with vitamin metabolism were thoroughly controlled. Serum concentrations were measured by commercial assays. The results were evaluated using Mann-Whitney's U-test and chi(2) analysis. RESULTS: OC-users showed significantly lower concentrations of cobalamin than controls. The rates of women with reduced, normal, and elevated levels differed significantly. Nine users but no control had frank cobalamin deficiency without clinical symptoms. Folate levels did not differ between the groups. Vegetarian diet, smoking or obesity did not have a significant influence. CONCLUSIONS: Routine measurement of cobalamin or folate in women using "low dose" OC is not warranted. Vitamin supplementation or different contraceptive methods should be considered in women with pre-existing cobalamin deficiency or restrictive dietary habits.

Eficacia y aceptabilidad en mujeres venezolanas de un anticonceptivo oral monofásico que contiene 20 microgramos de etinilestradiol y 150 microgramos de desogestrel / Efficacy and acceptability in Venezuelan females of a oral monophasic contraceptive containing ethynilestradiol 20 micrograms and desogestrel 150 micrograms

Evaluar la eficacia anticonceptiva, control de ciclo, aceptabilidad por parte de la usuaria, cambios en el peso corporal y tensión arterial con el uso de un anticonceptivo oral monofásico que contiene 20 microgramos de etilestradiol y 150 microgramos de desogestrel. Estudio abierto no comparativo multicéntrico donde participaron 122 mujeres proporcionando datos relativos a 600 ciclos. Ocho centros privados localizados en Caracas, Barquisimeto y Mérida. De las 122 mujeres evaluadas durante 600 ciclos, no se presentó embarazo, el control del ciclo tuvo una duración en todos los casos entre 3 a 5 y no huvo cambios estadísticamente significativos con respecto al peso corporal y tensión arterial. Los resultados derivados de este estudio sugieren que la reducción en la dósis de etinilestradiol a 20 microgramos permite todavía una máxima eficacia, buena aceptabilidad y un adecuado control de ciclo

Burnout, perceived stress, and cortisol responses to awakening.

OBJECTIVE: The effects of burnout and perceived stress on early morning free cortisol levels after awakening were investigated in a group of teachers. Previous studies revealed that cortisol levels show a significant increase after awakening, with high intraindividual stability. METHODS: Sixty-six teachers from local public schools (42 women and 24 men, mean age 42+/-5 years) were asked to sample saliva for cortisol analysis on 3 consecutive days. On each day, cortisol levels were measured at the time of awakening and 15, 30, and 60 minutes thereafter. On the night before the third day, subjects took 0.5 mg dexamethasone orally for testing glucocorticoid feedback inhibition. Burnout and perceived stress were measured by three different questionnaires. RESULTS: Perceived stress correlated with increases of cortisol levels during the first hour after awakening after dexamethasone pretreatment. In addition, teachers scoring high on burnout showed lower overall cortisol secretion on all sampling days, and a higher suppression of cortisol secretion after dexamethasone administration. In the subgroup of teachers with both high levels of perceived stress and high levels of burnout, a lower overall cortisol secretion was observed on the first 2 days, with stronger increases during the first hour after awakening after dexamethasone suppression. This subgroup also showed the lowest self-esteem, the highest external locus of control, and the highest number of somatic complaints. CONCLUSIONS: These results demonstrate differential effects of burnout and perceived stress on hypothalamic-pituitary-adrenal axis regulation.

[Oral contraception and the risk of breast cancer]. / Contraception orale et risque de cancer du sein.

The evaluation of carcinologic risks associated with various types of oral contraceptives remains unclear, because the constant evolution of composition and dosages in oestrogen and progestagen of pill. Since the oral contraceptives introduction 30 years ago, numerous epidemiological studies have analysed the association between OC and breast cancer risk. The recent meta-analysis of the Collaborative Group on Hormonal Factors in Breast Cancer provides large results. Original data from 54 studies representing about 90% of the published epidemiological studies were reanalyzed. The main findings are that is a small increase in the risk of having breast cancer diagnosed in current users of combined oral contraceptives and in women who had stopped use in past 10 years but that there is no evidence of an increase in the risk more than 10 years after stopping use. However, the cancers diagnosed in women who had used oral contraceptives are less advanced clinically than the cancers diagnosed in women who had not used them, suggesting a bias of screening in oral contraceptives users. However, the benefice-risk balance of OC is largely positive. The evaluation of third generation of pill is not yet available. Studies of third generation pill use and breast cancer risk are necessary.

Characterization of the novel progestin gestodene by receptor binding studies and transactivation assays.

Gestodene is a novel progestin used in oral contraceptives with an increased separation of progestogenic versus androgenic activity and a distinct antimineralocorticoid activity. This specific pharmacological profile of gestodene is defined by its pattern of binding affinities to a variety of steroid hormone receptors. In the present study the affinity of gestodene to the progesterone receptor (PR), the androgen receptor (AR), the glucocorticoid receptor (GR), the mineralocorticoid receptor (MR) and the estrogen receptor (ER) was re-evaluated by steroid binding assays and compared to those obtained for 3-keto-desogestrel and progesterone. The two synthetic progestins displayed identical high affinity to rabbit PR and similar marked binding to rat AR and GR, while progesterone showed high affinity to PR but only low binding to AR and GR. Furthermore, 3-keto-desogestrel exhibited almost no binding to MR, whereas gestodene, similar to progesterone, showed marked affinity to this receptor. In addition to receptor binding studies, transactivation assays were carried out to investigate the effects of gestodene on AR-, GR- and MR-mediated induction of transcription. In contrast to progesterone, which showed antiandrogenic activity, gestodene and 3-keto-desogestrel both exhibited androgenic activity. Furthermore, all three progestins exhibited weak GR-mediated antagonistic activity. In contrast to progesterone, which showed almost no glucocorticoid activity, gestodene and 3-keto-desogestrel showed weak glucocorticoid action. In addition, gestodene inhibited the aldosterone-induced reporter gene transcription, similar to progesterone, whereas unlike progesterone, gestodene did not induce reporter gene transcription. 3-Keto-desogestrel showed neither antimineralocorticoid nor mineralocorticoid action.

[Estrogens. Contraceptive therapy]. / Estrógenos. Terapia anticonceptiva.

To evaluate the competitive molecular phenomenon of Ethynyl-Estradiol (EE-2) from contraceptive formulations against the endogenous Estradiol (E-2) at the intra and the extracellular compartments, plasma and endometrial samples were simultaneously obtained on different days of the pseudomenstrual cycle from oral contraceptive users under EE-2+ Norgestrel (30 micrograms/+ 500 micrograms) and EE-2+ Norethindrone (50 micrograms + 1.0 mg) in order to quantify EE-2 & E-2. When measuring both molecules it was shown that the chronic administration of steroids regardless of the pharmacological action of the progestin component the lower content of EE-2 (30 micrograms) does not compete substantially at the circulating level permitting the cyclic fashion of the natural estradiol while at the endometrial compartment such phenomenon is not seen thus, a local infertility effect should be reconsidered to anticipate a different approach in the future of contraception.

PIP: 27 women participated in a study of systemic and endometrial estradiol concentrations in oral contraceptive (OC) users. 13 women aged 15-22 used OCs containing ethinyl estradiol 30 mcg and norgestrel 500 mg for 4-24 months, and 14 women aged 18-28 used OCs containing 50 mcg ethinyl estradiol and 1.0 mg norethindrone for 1-28 months. A peripheral blood sample and an endometrial biopsy were obtained at random from each woman on different cycle days to measure the concentration of endogenous estradiol. The ovarian response defined by endogenous plasma and endometrial estrogen levels differed greatly in the two groups. In the 30 mcg group, plasma estrogen levels were similar to those described in an ovulatory menstrual cycle, while this profile was not observed in the 50 mcg group. The endometrial estrogen profile described for women not using hormonal contraception was not observed in either of the two dose groups.

Estrógenos: terapia anticonceptiva / Estrogens: anticonceptive therapy

Para estudiar el fenómeno competitivo molecular del Etinil-Estradiol (EE-2) contenido en los anticonceptivos orales, en contra del estradiol (E-2) endógeno intra/extra-celular, muestras de plasma y endometrio fueron simultáneamente obtenidas en diferentes días del pseudociclo menstrual de mujeres bajo tratamiento con EE-2 + Norgestrel (30 ug + 500 ug) y EE2 + Noretindrona (50 ug + 1.0 mg). Los compuestos que contienen 30 ug de estrógeno sintético administrados crónicamente independientemente del contenido progestacional mostraron la no supresión del fenómeno cíclo hacia el 17 ß estradiol ovárico; ese fenómeno no se observó en el tejido endometrial. Un mecanismo local interceptivo debiera ser reconsiderado como un mecanismo farmacológico en el futuro de la regulación de la fertilidad