RESUMEN
The presence of estrogenic endocrine disruptors in aquatic environments has been a concern and bioassays are recommended tools for their monitoring. However, the physicochemical properties of contaminants and the environmental matrix features may influence the resultant response. This study aimed to assess this influence on the Yeast Estrogen Screen (YES) assay. Mixtures of 17ß-estradiol (E2) and humic acid (HA) were evaluated through the Schild approach aiming to investigate the interactions between estrogens and dissolved organic matter (DOM). Moreover, environmental samples from municipal landfill leachate and wastewater treatment plant (WWTP) influents and effluents were screened for (anti)estrogenic activity at both dissolved and particulate phases. Finally, results were statistically confronted with physicochemical parameters through principal component analysis (PCA). The HA test concentrations strongly reduced the E2 response, even at low levels. Humic substances may not only reduce estrogen bioavailability, but also interfere with the assay mechanism through enzymatic inhibition thus masking the sample estrogenic potential. Landfill leachate had total E2-Eq in the range 1282-2591 ng L-1, while WWTP influent and effluent were in the range 12.1-41.4 and Asunto(s)
Disruptores Endocrinos
, Contaminantes Químicos del Agua
, Contaminantes Químicos del Agua/análisis
, Materia Orgánica Disuelta
, Estrógenos/análisis
, Estradiol/análisis
, Disruptores Endocrinos/toxicidad
, Disruptores Endocrinos/análisis
, Estrona/análisis
, Antagonistas de Estrógenos/análisis
, Monitoreo del Ambiente/métodos
RESUMEN
Information on the occurrence and endocrine potencies of analogues of bisphenol A (BPA) and diglycidyl ester derivatives (BDGEs) of BPA and BPF is limited. Such information is, however, important as the current debate on BPA and the lowered BPA migration limit in Europe may provide an incentive for application of structural analogues. A new sensitive multi-analyte LC-ESI-MS/MS method was developed to measure 17 bisphenols (BPs) and 6 BDGEs in food, beverages and drinkware. Yeast based bioassays were used to determine the in vitro (anti)estrogenic and (anti)androgenic properties of these and 7 additional BPs and BDGEs. Drinkware of polycarbonate and other materials were analysed for BPs and BDGEs. Only BPA and BPS and both at trace levels were found in a few containers. A limited number of (canned) foods and beverages were also analysed. BPA was the most frequently detected BP (ranged from 0.03â¯ngâ¯mL-1 in a beverage sample to 68â¯ngâ¯g-1 in food). Other BPs detected were BPS, 2,2-BPF and 4,4-BPF. In addition BADGE, BADGE.HCl, BADGE.H2O and BADGE.2H2O were detected from 0.08â¯ngâ¯mL-1 in a beverage sample to 3.3â¯ngâ¯g-1 in food. In vitro testing showed that most BPs exhibited an equal or higher estrogenic potency than BPA and most of them also showed a higher anti-androgenic potency, i.e. BPB, BPCl, BPC, BPE, 4,4-BPF, BPP, BPAF, and BPTMC. Some BPs and BDGEs were not estrogenic, but showed an anti-estrogenic effect and were anti-androgenic too. BPS was only weakly estrogenic and BADGE.2H2O and BFDGE.2H2O showed no in vitro activity. The present data show that in addition to BPA, other BPs and BDGEs can be present in food and drinks, some displaying in vitro endocrine activities.
Asunto(s)
Antagonistas de Andrógenos/análisis , Compuestos de Bencidrilo/análisis , Ésteres/análisis , Antagonistas de Estrógenos/análisis , Contaminación de Alimentos/análisis , Fenoles/análisis , Compuestos de Bencidrilo/química , Cromatografía Liquida , Europa (Continente) , Fenoles/química , Espectrometría de Masas en Tándem/métodosRESUMEN
Complex tissue-specific and cell-specific signaling by the estrogen receptor (ER) frequently leads to the development of resistance to endocrine therapy for breast cancer. Pure ER antagonists, which completely lack tissue-specific agonist activity, hold promise for preventing and treating endocrine resistance, however an absence of structural information hinders the development of novel candidates. Here we synthesize a small panel of benzopyrans with variable side chains to identify pure antiestrogens in a uterotrophic assay. We identify OP-1074 as a pure antiestrogen and a selective ER degrader (PA-SERD) that is efficacious in shrinking tumors in a tamoxifen-resistant xenograft model. Biochemical and crystal structure analyses reveal a structure activity relationship implicating the importance of a stereospecific methyl on the pyrrolidine side chain of OP-1074, particularly on helix 12.
Asunto(s)
Antineoplásicos/farmacología , Benzopiranos/farmacología , Antagonistas de Estrógenos/farmacología , Neoplasias Mamarias Experimentales/tratamiento farmacológico , Pirrolidinas/farmacología , Fosfatasa Alcalina/análisis , Animales , Antineoplásicos/análisis , Antineoplásicos/síntesis química , Antineoplásicos/uso terapéutico , Benzopiranos/síntesis química , Benzopiranos/química , Benzopiranos/uso terapéutico , Proliferación Celular/efectos de los fármacos , Antagonistas de Estrógenos/análisis , Antagonistas de Estrógenos/síntesis química , Antagonistas de Estrógenos/uso terapéutico , Receptor alfa de Estrógeno/efectos de los fármacos , Femenino , Humanos , Células MCF-7 , Ratones Endogámicos BALB C , Ratones Desnudos , Conformación Proteica en Hélice alfa/efectos de los fármacos , Pirrolidinas/química , Pirrolidinas/uso terapéutico , Moduladores Selectivos de los Receptores de Estrógeno/análisis , Moduladores Selectivos de los Receptores de Estrógeno/síntesis química , Moduladores Selectivos de los Receptores de Estrógeno/farmacología , Estereoisomerismo , Útero/efectos de los fármacos , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Endocrine-disrupting chemicals are mainly discharged into the environment by wastewater treatment plants (WWTPs) and are known to induce adverse effects in aquatic life. Advanced treatment with ozone successfully removes such organic micropollutants, but an increase of estrogenic effects after the ozonation of hospital wastewater was observed in previous studies. In order to investigate this effect, estrogenic and androgenic as well as anti-estrogenic and anti-androgenic activities were observed during treatment of hospital wastewater using three different effect-based reporter gene bioassays. Despite different matrix influences, sensitivities, and test-specific properties, all assays used obtained comparable results. Estrogenic and androgenic activities were mainly reduced during the biological treatment and further removed during ozonation and sand filtration, resulting in non-detectable agonistic activities in the final effluent. An increased estrogenic activity after ozonation could not be observed in this study. Antagonistic effects were removed in the biological treatment by up to 50 % without further reduction in the advanced treatment. Due to the presence of antagonistic substances within the wastewater, masking effects were probable. Therefore, this study showed the relevance of antagonistic activities at hospital WWTPs and illustrates the need for a better understanding about antagonistic effects.
Asunto(s)
Antagonistas de Andrógenos/toxicidad , Bioensayo , Disruptores Endocrinos/toxicidad , Antagonistas de Estrógenos/toxicidad , Eliminación de Residuos Líquidos , Aguas Residuales/toxicidad , Contaminantes Químicos del Agua/toxicidad , Antagonistas de Andrógenos/análisis , Bioensayo/métodos , Disruptores Endocrinos/análisis , Antagonistas de Estrógenos/análisis , Estrógenos , Filtración , Ozono/química , Saccharomyces cerevisiae/efectos de los fármacos , Pruebas de Toxicidad , Eliminación de Residuos Líquidos/métodos , Aguas Residuales/química , Contaminantes Químicos del Agua/análisisRESUMEN
A considerable amount of scientific evidence indicates that a number of pharmaceuticals that could be detected in the environment can contribute towards the development of problems associated with human reproductive systems, as well as those of wildlife. We investigated the estrogenic and androgenic effects of select pharmaceuticals with high production volume and environmental relevance. We examined the receptor-binding activities of these pharmaceuticals in the T47D human cell line using altered secretion of cytokine CXCL12. Functional yeast-luciferase reporter gene assays were also employed to confirm the mechanism of receptor binding by estrogen and androgen. Non-steroidal anti-inflammatory drugs, namely ibuprofen, diclofenac and antiarrhythmic agent amiodarone showed strong anti-estrogenic effects in the T47D cell line. In the yeast-luciferase assay, these anti-inflammatory drugs also demonstrated anti-estrogenic potency and inhibited the E2 response in a concentration-dependent manner. Amiodarone did not exhibit any response in the yeast-luciferase assay; therefore, the endocrine disruption presumably occurred at a different level without directly involving the receptor. All the anti-inflammatory drugs considered in this study, including ketoprofen, naproxen and clofibrate, exhibited a dose-dependent antagonism towards the androgen receptor in the yeast-luciferase assays. Several other drugs, including the stimulant caffeine, did not show any response in the tests that were employed. A risk assessment analysis using 'Hazard Quotient' suggested a potential risk, especially in the cases of ibuprofen, ketoprofen, diclofenac and clofibrate. The results reveal the intrinsic endocrine disrupting nature of several pharmaceuticals and thus could contribute towards explaining a number of adverse health effects on humans and wildlife.
Asunto(s)
Andrógenos/análisis , Disruptores Endocrinos/análisis , Antagonistas de Estrógenos/análisis , Preparaciones Farmacéuticas/análisis , Receptores Androgénicos/metabolismo , Receptores de Estrógenos/antagonistas & inhibidores , Contaminantes Químicos del Agua/análisis , Andrógenos/toxicidad , Antiinflamatorios no Esteroideos/análisis , Antiinflamatorios no Esteroideos/toxicidad , Bioensayo/métodos , Línea Celular Tumoral , Quimiocina CXCL12/metabolismo , Relación Dosis-Respuesta a Droga , Disruptores Endocrinos/toxicidad , Estradiol/toxicidad , Antagonistas de Estrógenos/toxicidad , Genes Reporteros , Humanos , Luciferasas/genética , Saccharomyces cerevisiae/genética , Contaminantes Químicos del Agua/toxicidadRESUMEN
CYP2D6 is primarily responsible for the metabolism of clomiphene citrate (CC). The purpose of the present study was to investigate the relationship between CYP2D6 genotypes, concentrations of CC and its major metabolites and drug response in infertility patients. We studied 42 patients with ovulatory dysfunction treated with only CC. Patients received a dose of 100 mg/day CC on days 3-7 of the menstrual cycle. CYP2D6 genotyping and measurement of CC and the major metabolite concentrations were performed. Patients were categorized into CC responders or non-responders according to one cycle response for the ovulation. Thirty-two patients were CC responders and 10 patients were non-responders with 1 cycle treatment. The CC concentrations were highly variable within the same group, but non-responders revealed significantly lower (E)-clomiphene concentration and a trend of decreased concentrations of active metabolites compared to the responders. Nine patients with intermediate metabolizer phenotype were all responders. We confirmed that the CC and the metabolite concentrations were different according to the ovulation status. However, our results do not provide evidence for the contribution of CYP2D6 polymorphism to either drug response or CC concentrations.
Asunto(s)
Clomifeno/uso terapéutico , Citocromo P-450 CYP2D6/genética , Infertilidad/tratamiento farmacológico , Polimorfismo Genético , Adulto , Cromatografía Líquida de Alta Presión , Clomifeno/sangre , Clomifeno/metabolismo , Antagonistas de Estrógenos/análisis , Antagonistas de Estrógenos/metabolismo , Antagonistas de Estrógenos/uso terapéutico , Femenino , Genotipo , Humanos , Infertilidad/genética , Inducción de la Ovulación , Fenotipo , República de Corea , Espectrometría de Masas en TándemRESUMEN
In this study, for the first time, a coupled 1-mL microsyringe system was utilized to perform a miniaturized ionic liquid dispersive liquid-liquid microextraction (IL-DLLME) method. Danazol was extracted and determined via the developed method followed by micro-volume ultraviolet spectroscopy (UV). The extraction process was carried out by the injection of extraction solvent ionic liquid 1-octyl-3-methylimidazolium hexafluorophosphate [C8mimPF6] into sample solution (syringe A), and then rapid shoot the solution into syringe B. After that the shooting was repeated several times at a rate of 1 cycle/s. The extraction procedure was induced by the formation of cloudy solution, which was composed of fine drops of [C8mimPF6] dispersed entirely into sample solution with the help of shooting without any dispersive solvent, ultrasonication or high temperature. Several important parameters affecting the extraction efficiency were studied and optimized. Under the optimized conditions, the limit of detection (LOD) was 0.055 µg/mL (capsule) or 0.054 µg/mL (serum) at a signal-to-noise ratio of 3. The calibration curve was linear over the range of 0.62-25 µg/mL. The proposed method was successfully applied to danazol capsule and the real mice serum samples and good spiked recoveries in the range of 90.5-103.4% were obtained. The obtained results of this work were in good agreement with the results of HPLC.
Asunto(s)
Danazol/sangre , Antagonistas de Estrógenos/sangre , Líquidos Iónicos/química , Microextracción en Fase Líquida/instrumentación , Espectrofotometría Ultravioleta/instrumentación , Animales , Cápsulas , Danazol/análisis , Diseño de Equipo , Antagonistas de Estrógenos/análisis , Límite de Detección , Ratones , JeringasRESUMEN
CYP2D6 is primarily responsible for the metabolism of clomiphene citrate (CC). The purpose of the present study was to investigate the relationship between CYP2D6 genotypes, concentrations of CC and its major metabolites and drug response in infertility patients. We studied 42 patients with ovulatory dysfunction treated with only CC. Patients received a dose of 100 mg/day CC on days 3-7 of the menstrual cycle. CYP2D6 genotyping and measurement of CC and the major metabolite concentrations were performed. Patients were categorized into CC responders or non-responders according to one cycle response for the ovulation. Thirty-two patients were CC responders and 10 patients were non-responders with 1 cycle treatment. The CC concentrations were highly variable within the same group, but non-responders revealed significantly lower (E)-clomiphene concentration and a trend of decreased concentrations of active metabolites compared to the responders. Nine patients with intermediate metabolizer phenotype were all responders. We confirmed that the CC and the metabolite concentrations were different according to the ovulation status. However, our results do not provide evidence for the contribution of CYP2D6 polymorphism to either drug response or CC concentrations.
Asunto(s)
Adulto , Femenino , Humanos , Cromatografía Líquida de Alta Presión , Clomifeno/sangre , Citocromo P-450 CYP2D6/genética , Antagonistas de Estrógenos/análisis , Genotipo , Infertilidad/tratamiento farmacológico , Inducción de la Ovulación , Fenotipo , Polimorfismo Genético , República de Corea , Espectrometría de Masas en TándemRESUMEN
OBJECTIVE: Adverse health outcomes related to exposure to endocrine disrupting chemicals, including increased incidences of coronary heart disease, prostate and testicular cancers, and congenital disabilities, have been reported in firefighters or their offspring. We, therefore, measured the estrogenic and antiestrogenic activity of extracts of used firefighter gear to assess exposure to these agents. METHODS: Extracts and known chemical contaminants were examined for estrogenicity and antiestrogenicity in yeast cells expressing the estrogen receptor. RESULTS: Most extracts of used gear and phthalate diesters detectable on this gear displayed strong antiestrogenic effects. Notably, new glove and hood extracts showed significant estrogenic activity. CONCLUSIONS: Overall, our data suggest that firefighters are exposed to both estrogenic and antiestrogenic agents, possibly phthalates that may lead to health risks observed in this occupation as a result of perturbation of hormone homeostasis.
Asunto(s)
Disruptores Endocrinos/análisis , Antagonistas de Estrógenos/análisis , Estrógenos/análisis , Bomberos , Exposición Profesional/análisis , Ropa de Protección , HumanosRESUMEN
OBJECTIVE: To establish gene assays for determining (anti) estrogen effect of environmental chemicals; and to compare the reactivity and sensitivity of two assays with different estrogen subtype. METHODS: Human estrogen receptor a (hERalpha) and hERbeta mediated reporter gene assays employing firefly luciferase (Luc) were developed. The expression plasmid hERalpha or hERbeta was constructed and transiently co-transfected into LLC-MK2 cells with pERE-minP-Luc2P reporter plasmid and the control plasmid pGL4.74. Estradiol (E2) and diethylstilbestrol (DES) served as positive test substances to verify the performance of the assays. The effectiveness of the assays for detecting anti-estrogenic activity was tested using 10(-5) mol/L ICI 182, 780 under different concentrations of E2. The performance of the two subtype-mediated assays was verified and compared using bisphenol A (BPA) and genistein (GS). RESULTS: The hERalpha mediated assay found expression of reported gene at 1.9 x 10(-11) mol/L E2; and the largest luciferase activity was shown at 10(-8) mol/L E2, resulting in 30.7-fold of vehicle control. The hERbeta mediated assay found expression of reporter gene at 2.2 x 10(11) mol/L E2, and the largest luciferase activity was shown at 10(-8) mol/L E2, resulting in 14.4-fold of vehicle control. ICI 182, 780 inhibited estrogenic activity of E2 significantly. In both assays, E2 failed to induce luciferase activity without hER-pcDNA3.1. BPA and GS induced luciferase activity. CONCLUSION: Both assays have high sensitivity and reproducibility for detecting (anti) estrogen effect. The pGL4-based hERbeta has lower sensitivity than the hERalpha- mediated reporter gene assay. BPA shows stronger estrogenic activity than GS in hERalpha mediated reporter gene assay; whereas, GS shows stronger estrogenic activity than BPA in hERbeta mediated reporter gene assay.
Asunto(s)
Antagonistas de Estrógenos/análisis , Genes Reporteros , Receptores de Estrógenos/genética , Compuestos de Bencidrilo/química , Línea Celular , Dietilestilbestrol/antagonistas & inhibidores , Estradiol/química , Genisteína/química , Humanos , Luciferasas de Luciérnaga , Fenoles/química , Plásmidos , Reproducibilidad de los Resultados , TransfecciónRESUMEN
Use of personal care products is widespread in the United States but tends to be greater among African Americans than whites. Of special concern is the possible hazard of absorption of chemicals with estrogenic activity (EA) or anti-EA (AEA) in these products. Such exposure may have adverse health effects, especially when it occurs during developmental windows (e.g., prepubertally) when estrogen levels are low. We assessed the ethanol extracts of eight commonly used hair and skin products popular among African Americans for EA and AEA using a cell proliferation assay with the estrogen sensitive MCF-7:WS8 cell line derived from a human breast cancer. Four of the eight personal care products tested (Oil Hair Lotion, Extra-dry Skin Lotion, Intensive Skin Lotion, Petroleum Jelly) demonstrated detectable EA, whereas three (Placenta Hair Conditioner, Tea-Tree Hair Conditioner, Cocoa Butter Skin Cream) exhibited AEA. Our data indicate that hair and skin care products can have EA or AEA, and suggest that laboratory studies are warranted to investigate the in vivo activity of such products under chronic exposure conditions as well as epidemiologic studies to investigate potential adverse health effects that might be associated with use of such products.
Asunto(s)
Proliferación Celular/efectos de los fármacos , Cosméticos/farmacología , Antagonistas de Estrógenos/farmacología , Estrógenos/farmacología , Cabello , Cuidados de la Piel , Negro o Afroamericano , Antagonistas de Estrógenos/análisis , Estrógenos/análisis , Humanos , Células MCF-7 , Medición de Riesgo , Estados UnidosRESUMEN
Thousands of environmental chemicals are subject to regulatory review for their potential to be endocrine disruptors (ED). In vitro high-throughput screening (HTS) assays have emerged as a potential tool for prioritizing chemicals for ED-related whole-animal tests. In this study, 1814 chemicals including pesticide active and inert ingredients, industrial chemicals, food additives, and pharmaceuticals were evaluated in a panel of 13 in vitro HTS assays. The panel of in vitro assays interrogated multiple end points related to estrogen receptor (ER) signaling, namely binding, agonist, antagonist, and cell growth responses. The results from the in vitro assays were used to create an ER Interaction Score. For 36 reference chemicals, an ER Interaction Score >0 showed 100% sensitivity and 87.5% specificity for classifying potential ER activity. The magnitude of the ER Interaction Score was significantly related to the potency classification of the reference chemicals (p < 0.0001). ERα/ERß selectivity was also evaluated, but relatively few chemicals showed significant selectivity for a specific isoform. When applied to a broader set of chemicals with in vivo uterotrophic data, the ER Interaction Scores showed 91% sensitivity and 65% specificity. Overall, this study provides a novel method for combining in vitro concentration response data from multiple assays and, when applied to a large set of ER data, accurately predicted estrogenic responses and demonstrated its utility for chemical prioritization.
Asunto(s)
Disruptores Endocrinos/análisis , Receptor alfa de Estrógeno/agonistas , Receptor beta de Estrógeno/agonistas , Ensayos Analíticos de Alto Rendimiento , Modelos Químicos , Algoritmos , Animales , Bioensayo , Antagonistas de Estrógenos/análisis , Receptor alfa de Estrógeno/antagonistas & inhibidores , Receptor beta de Estrógeno/antagonistas & inhibidores , Estrógenos/análisis , Humanos , Células MCF-7 , Plaguicidas , Transducción de SeñalRESUMEN
Endocrine-disrupting chemicals are exogenous substances that alter the function of the endocrine system, with adverse health effects on organisms or their progeny. In vitro estrogen receptor (ER) reporter gene assays have long been used to measure estrogenic activity in wastewater. Nevertheless, there is still uncertainty about their usefulness in environmental monitoring on account of a discrepancy between the estrogenic response of the in vitro assay and concentrations of estrogenic compounds determined by chemical analysis. Here, we measured estrogenic and antiestrogenic activities in wastewater by ERα reporter gene assay. All samples were simultaneously analyzed for estrone, 17ß-estradiol, estriol, and 17α-ethynylestradiol, and the concentrations were used to predict estrogenic activity. All samples in which measured estrogenic activity was significantly lower than predicted showed strong antiestrogenic activity. In addition, we confirmed that the fraction that did not have antiestrogenic activity showed stronger estrogenic activity than the unfractionated wastewater extract. These results indicate that antiestrogenic compounds in wastewater suppress the activity of natural estrogens, and the reporter gene assay represents the net activity.
Asunto(s)
Disruptores Endocrinos/toxicidad , Congéneres del Estradiol/toxicidad , Antagonistas de Estrógenos/toxicidad , Receptor alfa de Estrógeno/genética , Aguas Residuales/química , Contaminantes Químicos del Agua/toxicidad , Animales , Disruptores Endocrinos/análisis , Monitoreo del Ambiente/métodos , Congéneres del Estradiol/análisis , Antagonistas de Estrógenos/análisis , Células HEK293 , Humanos , Oryzias , Contaminantes Químicos del Agua/análisisRESUMEN
Anti-estrogenic activity in wastewater is gaining increased attention because of its endocrine-disrupting function. In this study, the level and removal efficiency by coagulation of anti-estrogenic activity in secondary effluents of domestic wastewater treatment plants were studied. Anti-estrogenic activity was detected in secondary effluent samples at a tamoxifen (TAM) equivalent concentration level of 0.38-0.94 mg-TAML(-1). Dissolved organic matters (DOM) with the molecular weight (MW) less than 3000 Da in hydrophobic acids (HOA) and hydrophobic neutrals (HON) fractions of the secondary effluent were the key fractions related to anti-estrogenic activity. Coagulation with FeCl(3) and polyaluminium chloride (PAC) can remove the anti-estrogenic activity of the secondary effluents, but the removal efficiency was limited. The removal efficiency using FeCl(3) coagulant was higher than that induced by PAC. Dissolved organic carbon was continuously removed with increased coagulant dose (0-120 mg L(-1) FeCl(3) or 0-60 mg L(-1) PAC). However, the removal of anti-estrogenic activity was not enhanced further when the coagulant concentration was beyond a critical value (30 mg L(-1) FeCl(3) or 10 mg L(-1) PAC). The highest removal of anti-estrogenic activity was about 36% by FeCl(3) and 20% by PAC. Size exclusion chromatography results indicated difficulty in removing DOM with MW less than 3000 Da in the secondary effluent during coagulation even at a high coagulant concentration, which led to low removal efficiency of anti-estrogenic activity.
Asunto(s)
Antagonistas de Estrógenos/análisis , Eliminación de Residuos Líquidos/métodos , Aguas Residuales/química , Contaminantes Químicos del Agua/análisis , Purificación del Agua/métodosRESUMEN
Adaptive or compensatory responses to chemical exposure can significantly influence in vivo concentration-duration-response relationships. This study provided data to support development of a computational dynamic model of the hypothalamic-pituitary-gonadal axis of a model vertebrate and its response to aromatase inhibitors as a class of endocrine active chemicals. Fathead minnows (Pimephales promelas) were either exposed to the aromatase inhibitor fadrozole (0.5 or 30 µg/l) continuously for 1, 8, 12, 16, 20, 24, or 28 days or exposed for 8 days and then held in control water (no fadrozole) for an additional 4, 8, 12, 16, or 20 days. The time course of effects on ovarian steroid production, circulating 17ß-estradiol (E2) and vitellogenin (VTG) concentrations, and expression of steroidogenesis-related genes in the ovary was measured. Exposure to 30 µg fadrozole/l significantly reduced plasma E2 and VTG concentrations after just 1 day and those effects persisted throughout 28 days of exposure. In contrast, ex vivo E2 production was similar to that of controls on day 8-28 of exposure, whereas transcripts coding for aromatase and follicle-stimulating hormone receptor were elevated, suggesting a compensatory response. Following cessation of fadrozole exposure, ex vivo E2 and plasma E2 concentrations exceeded and then recovered to control levels, but plasma VTG concentrations did not, even after 20 days of depuration. Collectively these data provide several new insights into the nature and time course of adaptive responses to an aromatase inhibitor that support development of a computational model (see companion article).
Asunto(s)
Adaptación Fisiológica/efectos de los fármacos , Inhibidores de la Aromatasa/toxicidad , Cyprinidae/fisiología , Antagonistas de Estrógenos/toxicidad , Fadrozol/toxicidad , Sistema Hipotálamo-Hipofisario/efectos de los fármacos , Ovario/efectos de los fármacos , Alternativas a las Pruebas en Animales , Animales , Inhibidores de la Aromatasa/análisis , Estradiol/sangre , Antagonistas de Estrógenos/análisis , Fadrozol/análisis , Femenino , Sistema Hipotálamo-Hipofisario/enzimología , Masculino , Ovario/enzimología , Valor Predictivo de las Pruebas , Factores de Tiempo , Vitelogeninas/sangreRESUMEN
Estrogen receptor (ER) modulators are a serious health issue but estrogenic compounds, especially antagonists of ER function, are widely screened for in search of novel therapeutics against hormonal diseases such as the breast cancer. Here we report a novel and a simple bioassay for estrogenic and anti-estrogenic compounds based on ligand-dependent recruitment of ER co-activator steroid receptor co-activator 1 (SRC-1) to purified Renilla luciferase-tagged ERα. In this assay, in vivo-biotinylated (E. coli) SRC-1, purified Renilla luciferase-ERα, and the analyte sample are mixed and incubated for 2 h in a streptavidin-coated microtiter wells, and after one washing step, luminescence is measured with a simple instrument. The assay does not require chemical labeling of the components and shows good sensitivity (25 pM E(2)) and wide dynamic range of more than four orders of magnitude.
Asunto(s)
Bioensayo/métodos , Disruptores Endocrinos/análisis , Antagonistas de Estrógenos/análisis , Estrógenos/análisis , Luciferasas de Renilla/metabolismo , Relación Dosis-Respuesta a Droga , Disruptores Endocrinos/metabolismo , Escherichia coli/genética , Estradiol/análisis , Estradiol/metabolismo , Antagonistas de Estrógenos/metabolismo , Receptor alfa de Estrógeno/química , Receptor alfa de Estrógeno/genética , Receptor alfa de Estrógeno/metabolismo , Estrógenos/metabolismo , Humanos , Luciferasas de Renilla/química , Luciferasas de Renilla/genética , Coactivador 1 de Receptor Nuclear/química , Coactivador 1 de Receptor Nuclear/metabolismo , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Sensibilidad y EspecificidadRESUMEN
Flavonoids such as naringenin and morin are ubiquitous in a wide range of foods isolated from plants, and have diverse effects on plants even on human health. Here, we establish a selective visual method for recognition of aringenin and morin based on the "switched on" fluorescence induced by a metal-organic coordination polymer of Zn(bix) [bix=1,4-bis(imidazol-1-ylmethyl)benzene]. Owing to the coordination interaction of aringenin and morin with Zn(II) from the polymeric structure of Zn(bix), the conformational free rotation of naringenin and morin is restricted leading to relatively rigid structures. And as a consequence, the fluorescence is switched on. While luteolin and quercetin, holding a very similar structure with naringenin and morin, have no such fluorescence enhancement most likely owing to the 3'-hydroxy substitution in the B ring. Under 365 nm UV lamp light, we can visually recognize and discriminate naringenin and morin from them each other and luteolin as well as quercetin based on the colors of their emission. With this recognition system, the detection of naringenin and morin in human urine was made with satisfactory results.
Asunto(s)
Derivados del Benceno/química , Antagonistas de Estrógenos/orina , Flavanonas/orina , Flavonoides/orina , Compuestos Organometálicos/química , Zinc/química , Antioxidantes/análisis , Antagonistas de Estrógenos/análisis , Flavanonas/análisis , Flavonoides/análisis , Humanos , Sensibilidad y Especificidad , Espectrometría de Fluorescencia/métodosRESUMEN
Wastewaters from various industries are a main source of the contaminants in aquatic environments. The authors evaluated the hormonal activities (estrogenic/anti-estrogenic activities, androgenic/anti-androgenic activities) and genotoxicity of various effluents from textile and dyeing plants, electronic and electroplate factories, pulp and paper mills, fine chemical factories, and municipal wastewater treatment plants in the Pearl River Delta region by using in vitro bioassays (yeast estrogen screen [YES]; yeast androgen screen [YAS]; and genotoxicity assay [umu/SOS]) combined with chemical analysis. The results demonstrated the presence of estrogenic, anti-estrogenic, and anti-androgenic activity in most industrial effluents, whereas no androgenic activities were detected in all of the effluents. The measured estrogenic activities expressed as estradiol equivalent concentrations (EEQs) ranged from below detection (3 of 26 samples) to 40.7 ng/L, with a mean of 7.33 ng/L in all effluents. A good linear relationship was found between the EEQs measured by YES bioassay and the EEQs calculated from chemical concentrations. These detected estrogenic compounds, such as 4-nonylphenol and estrone, were responsible for the estrogenic activities in the effluents. The genotoxic effects expressed as benzo[a]pyrene equivalent concentrations (BaP EQs) varied between below detection and 88.2 µg/L, with a mean of 8.76 µg/L in all effluents. The target polycyclic aromatic hydrocarbons were minor contributors to the genotoxicity in the effluents, and some nontarget compounds in the effluents were responsible for the measured genotoxicity. In terms of estrogenic activities and genotoxicity, discharge of these effluents could pose high risks to aquatic organisms in the receiving environments.
Asunto(s)
Monitoreo del Ambiente/métodos , Estrógenos/toxicidad , Aguas Residuales/química , Contaminantes Químicos del Agua/toxicidad , Andrógenos/análisis , Andrógenos/toxicidad , Bioensayo , China , Estradiol/análisis , Estradiol/toxicidad , Antagonistas de Estrógenos/análisis , Antagonistas de Estrógenos/toxicidad , Estrógenos/análisis , Estrona/análisis , Estrona/toxicidad , Mutágenos/análisis , Mutágenos/toxicidad , Fenoles/análisis , Fenoles/toxicidad , Hidrocarburos Policíclicos Aromáticos/análisis , Hidrocarburos Policíclicos Aromáticos/toxicidad , Ríos/química , Contaminantes Químicos del Agua/análisisRESUMEN
Tomato fruit has assumed the status of 'functional food' due to the association between its consumption and a reduced likelihood of certain types of cancers and CVD. The nutraceutical value of tomatoes can be affected by the cultivation conditions, e.g. the phytochemical content of the fruits may increase with the establishment of beneficial mycorrhizal symbioses in the plants. A multidisciplinary study was carried out to gain knowledge on the antioxidant, oestrogenic/anti-oestrogenic and genotoxic activity of tomato fruits produced by mycorrhizal plants. The present results showed that the symbiosis positively affected the growth and mineral nutrient content of tomato plants and enhanced the nutritional and nutraceutical value of tomato fruits through modifications of plant secondary metabolism, which led to increased levels of lycopene in fruits obtained from mycorrhizal plants, compared with controls. Moreover, such changes did not result in the production of mutagenic compounds, since tomato extracts induced no in vitro genotoxic effects. Fruit extracts, both hydrophilic and the lipophilic fractions, originating from mycorrhizal plants strongly inhibited 17-ß-oestradiol-human oestrogen receptor binding, showing significantly higher anti-oestrogenic power compared with controls. The present study shows that beneficial plant symbionts, such as mycorrhizal fungi, can lead to the production of safe and high-quality food, which is an important societal issue strongly demanded by both consumers and producers.
Asunto(s)
Frutas/química , Frutas/microbiología , Alimentos Funcionales/análisis , Alimentos Funcionales/microbiología , Micorrizas/fisiología , Solanum lycopersicum/química , Solanum lycopersicum/microbiología , Antioxidantes/análisis , Suplementos Dietéticos/efectos adversos , Suplementos Dietéticos/análisis , Antagonistas de Estrógenos/análisis , Antagonistas de Estrógenos/farmacología , Receptor alfa de Estrógeno/antagonistas & inhibidores , Receptor alfa de Estrógeno/genética , Receptor alfa de Estrógeno/metabolismo , Frutas/efectos adversos , Frutas/crecimiento & desarrollo , Alimentos Funcionales/efectos adversos , Humanos , Concentración de Iones de Hidrógeno , Solanum lycopersicum/efectos adversos , Solanum lycopersicum/crecimiento & desarrollo , Masculino , Minerales/análisis , Mutágenos/análisis , Mutágenos/farmacología , Micorrizas/química , Valor Nutritivo , Fitoestrógenos/análisis , Fitoestrógenos/farmacología , Extractos Vegetales/análisis , Extractos Vegetales/farmacología , Control de Calidad , Proteínas Recombinantes/antagonistas & inhibidores , Proteínas Recombinantes/metabolismo , Elementos de Respuesta/efectos de los fármacos , SimbiosisRESUMEN
Gunnera perpensa (Gunneraceae) is an African plant widely used in traditional medicine. This species is known for its activity involving the female reproductive system, such as inducing or increasing labor, treating female infertility, expelling the placenta and/or preventing post-partum hemorrhage. These properties are probably due to (z)-venusol, a majoritary compound, and its action in conjunction with substances in the whole extract and other natural products. In southern Brazil, a native species Gunnera manicata L. that also belongs to Gunneraceae can be found. In spite of the traditional use of G. perpensa, there is no pharmacological and phytochemical information regarding the South American Gunnera species. Therefore, the aim of this study was to investigate the activity of Brazilian G. manicata aqueous extracts on the reproductive system of immature female Wistar rats through a uterotrophic assay and to verify the presence of (z)-venusol by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Data were analyzed by analysis of variance (ANOVA) and Bonferroni´s post-hoc test (p< 0.01). Results obtained shown that G. manicata extracts did not present in vivo anti or estrogenic activity. Furthermore, (z)-venusol compound was not found. This study represents the first preliminary screening done on the South American G. manicata species.
Gunnera perpensa (Gunneraceae) é uma planta de origem africana extensamente utilizada na medicina tradicional do país. Esta espécie é conhecida por suas atividades no sistema reprodutor feminino, como indução ou aumento do trabalho de parto, tratamento da infertilidade em mulheres, expulsão da placenta e/ou impedimento de hemorragia pós-parto. Tais atividades devem-se, provavelmente, ao sinergismo existente entre o (z)-venusol, composto majoritário, e outros compostos presentes na planta. No sul do Brasil, encontra-se uma espécie nativa, Gunnera manicata L., pertencente à família Gunneraceae. Apesar do uso tradicional de G. perpensa, não há informações farmacológicas e fitoquímicas a respeito da espécie sul Americana de Gunnera. Assim, o objetivo deste estudo foi investigar a atividade de extratos aquosos da espécie brasileira G. manicata no sistema reprodutor de ratas Wistar imaturas através de ensaio uterotrófico e verificar a presença do composto (z)-venusol utilizando-se cromatografia líquida acoplada a espectrômetro de massas em tandem (CL-EM/EM). Para a análise estatística, utilizou-se ANOVA/Bonferroni (p<0,01). Os resultados obtidos demonstraram que os extratos de G. manicata testados não apresentaram atividade anti ou estrogênica in vivo. Na análise química não foi verificada a presença do composto (z)-venusol. Este estudo representa o primeiro screening realizado com a espécie sul-americana G. manicata.