Time - sensitive effects of Zhuang medicated thread moxibustion on estrogen level in female perimenopausal model rats / Efectos sensibles al tiempo de la moxibustión con hilo medicado de Zhuang sobre el nivel de estrógeno en ratas modelo perimenopáusicas hembras

The present study was conducted to ascertain the estrogenic effect of Zhuang Medicated Thread Moxibustion (ZMTM) and explore its time - sensitive impact on estradiol (E2) in female perimenopausal rats. 40 female rats were randomized into four gr oups: the control, model, ZMTM, and acupuncture groups. The perimenopausal syndrome was induced in the last three groups with a daily subcutaneous dose of 80 mg/kg of 4 - vinylcyclohexene diepoxide for 15 days. Afterward, rats in the model and control group s were fed routinely, while rats in the ZMTM and acupuncture groups were treated with six ZMTM and acupuncture courses, respectively. Results of the study suggested that following the six courses of treatment, the E2 level in the model group was significan tly the lowest, while the regular group was the highest (P < 0.05). There was also a gradual increase in the E2 level of the ZMTM group compared to the model and acupuncture groups, e.g. after the 5th and 6th courses of treatment, their E2 level was signif icantly higher than the model and acupuncture groups. The ZMTM group was better than the model and acupuncture groups. In summary, ZMTM can improve perimenopausal induced rats' estrogen level.

El presen te estudio se llevó a cabo para determinar el efecto estrogénico de la moxibustión con hilo medicado Zhuang (ZMTM) y explorar su impacto sensible al tiempo en el estradiol (E2) en ratas hembras perimenopáusicas. Se dividió al azar una muestra de 40 ratas h embras en cuatro grupos: control, modelo, ZMTM y acupuntura. El síndrome perimenopáusico se indujo en los últimos tres grupos con una dosis subcutánea diaria de 80 mg/kg de diepóxido de 4 - vinilciclohexeno durante 15 días. Después, las ratas en los grupos m odelo y control fueron alimentadas rutinariamente, mientras que las ratas en los grupos ZMTM y acupuntura recibieron seis cursos de ZMTM y acupuntura, respectivamente. Los resultados del estudio sugieren que después de los seis cursos de tratamiento, el ni vel de E2 en el grupo modelo fue significativamente más bajo, mientras que el grupo regular fue más alto ( p < 0,05). También hubo un aumento gradual en el nivel de E2 del grupo ZMTM en comparación con los grupos modelo y acupuntura, por ejemplo, desp ués del quinto y sexto cursos de tratamiento, su nivel de E2 fue significativamente más alto que los grupos modelo y acupuntura. El grupo ZMTM fue mejor que los grupos modelo y acupuntura. En resumen, el ZMTM puede mejorar el nivel de estrógeno de las rata s inducidas por la perimenopausia.

Endocrine-disrupting substances: I. Relative risks of PFAS in drinking water.

Concentrations of per and polyfluorinated alkyl substances (PFAS) in drinking water are significantly lower than in vivo levels of the native target hormone. These concentrations are orders of magnitude lower than the hormone in question, particularly when corrected for transactivation. A pregnant woman can excrete about 7,000 µg/day of total estrogens. A low-dose oral contraceptive pill contains 20 µg estradiol. Soy-based baby formula contains phytoestrogens equivalent to a low-dose oral contraceptive pill. A woman on a low-dose oral hormone replacement therapy consumes about 0.5-2 mg/day of one or more estrogens. The levels of endocrine-disrupting substances (EDSs) exposure by oral, respiratory, or dermal routes have the potential to make removing PFAS from drinking water due to its estrogenic activity divert valuable resources. These levels become even less of a threat when their estrogenic potencies are compared with those of the target hormones present as contaminants in water and even more so when compared with levels commonly present in human tissues. The fact that PFAS constitute a tiny fraction compared to exposure to phytoestrogens makes the effort even more insignificant. If PFAS are to be removed from drinking water, it is not due to their estrogenic activity.

Distribution, ecological fate, and risks of steroid estrogens in environmental matrices.

Over the past two decades, steroidal estrogens (SEs) such as 17α-ethylestradiol (EE2), 17ß-estradiol (E2),17α-estradiol (17α-E2), estriol (E3) and estrone (E1) have elicited worldwide attention due to their potentially harmful effects on human health and aquatic organisms even at low concentration ng/L. Natural steroidal estrogens exhibit greater endocrine disruption potency due to their high binding effect on nuclear estrogen receptors (ER). However, less has been explored regarding their associated environmental risks and fate. A comprehensive bibliometric study of the current research status of SEs was conducted using the Web of Science to assess the development trends and current knowledge of SEs in the last two decades, from 2001 to 2021 October. The number of publications has tremendously increased from 2003 to 2021. We summarized the contamination status and the associated ecological risks of SEs in different environmental compartments. The results revealed that SEs are ubiquitous in surface waters and natural SEs are most studied. We further carried out an in-depth evaluation and synthesis of major research hotspots and the dominant SEs in the matrices were E1, 17ß-E2, 17α-E2, E3 and EE2. Nonetheless, investigations of SEs in soils, groundwater, and sediments remain scarce. This study elucidates SEs distribution, toxicological risks, ecological fate and mitigation measures, which will be beneficial for future monitoring, management, and risk assessment. Further studies are recommended to assess the toxicological risks of different SEs in complex environmental matrices to pursue a more precise and holistic quantitative estimation of estrogenic risk.

Estrogenic and retinoid-like activity in stagnant waters with mass occurrence of water blooms.

Stagnant freshwaters can be affected by anthropogenic pollution and eutrophication that leads to massive growth of cyanobacteria and microalgae forming complex water blooms. These can produce various types of bioactive compounds, some of which may cause embryotoxicity, teratogenicity, endocrine disruption and impair animal or human health. This study focused on potential co-occurrence of estrogenic and retinoid-like activities in diverse stagnant freshwaters affected by phytoplankton blooms with varying taxonomic composition. Samples of phytoplankton bloom biomass and its surrounding water were collected from 17 independent stagnant water bodies in the Czech Republic and Hungary. Total estrogenic equivalents (EEQ) of the most potent samples reached up to 4.9 ng·g-1 dry mass (dm) of biomass extract and 2.99 ng·L-1 in surrounding water. Retinoic acid equivalent (REQ) measured by in vitro assay reached up to 3043 ng·g-1 dm in phytoplankton biomass and 1202 ng·L-1in surrounding water. Retinoid-like and estrogenic activities at some sites exceeded their PNEC and effect-based trigger values, respectively. The observed effects were not associated with any particular species of cyanobacteria or algae dominating the water blooms nor related to phytoplankton density. We found that taxonomically diverse phytoplankton communities can produce and release retinoid-like compounds to surrounding water, while estrogenic potency is likely related to estrogens of anthropogenic origin adsorbed to phytoplankton biomass. Retinoids occurring in water blooms are ubiquitous signalling molecules, which can affect development and neurogenesis. Selected water bloom samples (both water and biomass extracts) with retinoid-like activity caused effects on neurodifferentiation in vitro corresponding to those of equivalent all-trans-retinoic acid concentrations. Co-occurrence of estrogenic and retinoid-like activities in stagnant water bodies as well as the potential of compounds produced by water blooms to interfere with neural differentiation should be considered in the assessment of risks associated with water blooms, which can comprise complex mixtures of natural and anthropogenic bioactive compounds.

Rapid Detection of Five Estrogens Added Illegally to Dietary Supplements by Combining TLC with Raman Imaging Microscope.

Estrogens added illegally to dietary supplements are hazardous to human health. Traditional detection and analysis methods have many limitations, and we have developed an assay that combines thin-layer chromatography with Raman imaging microscopy (TLC-RIM). The five estrogens (estrone, estradiol, estriol, ethinyl estradiol, and diethylstilbestrol) were initially separated by TLC, then detected by area scanning Raman imaging with a 532 nm laser under a microscope. Raman spectra were obtained for each estrogen, which were used for detecting estrogen illegally added to botanical dietary supplements. The LOD of each estrogen was 0.4, 1.0, 0.8, 0.2, and 0.2 mg/mL, respectively. The matrix in the real sample did not interfere with the detection of estrogens. The method was fast, sensitive, stable, specific, and reliable.

Secondary treatment phase of tertiary wastewater treatment works significantly reduces estrogenic load.

Estrogenic compounds enter waterways via effluents from wastewater treatment works (WWTW), thereby indicating a potential risk to organisms inhabiting adjacent receiving waters. However, little is known about the loads or concentrations of estrogenic compounds that enter Australian WWTWs, the efficiency of removing estrogenic compounds throughout the various stages of tertiary WWTW processes (which are common in Australia), nor the concentrations released into estuarine or marine receiving waters, and the associated risk for aquatic taxa residing in these environments. Therefore, seven estrogenic compounds, comprising the natural estrogens estrone (E1), 17ß-estradiol (E2) and estriol (E3), the synthetic estrogen (EE2), and the industrial chemicals bisphenol A (BPA), 4-t-octyl phenol (4-t-OP) and 4-nonyl phenol (4-NP), in wastewater samples were quantified via liquid chromatographic-mass spectrometry (LC-MS) after solid-phase extraction at different stages of wastewater treatment and associated receiving waters. The concentrations of the target compounds in wastewater ranged from < LOQ (limit of quantification) to 158 ng/L for Tanilba Bay WWTW and < LOQ to 162 ng/L for Belmont WWTW. Most target compounds significantly declined after the secondary treatment phase. Appreciable removal efficiency throughout the treatment process was observed with removal from 39.21 to 99.98% of influent values at both WWTWs. The reduction of the natural estrogens (E1, E2 and E3) and 4-t-OP were significantly greater than EE2, BPA, and 4-NP in both WWTWs. Risk quotients (RQs) were calculated to assess potential ecological risks from individual estrogenic compounds. In predicted diluted effluents, no targeted compounds showed any ecological risk (RQ ≤1.65 × 10-2) at both WWTWs. Similarly, all RQs for shore samples at both WWTWs were below 1. Finally, the hazard index (HI), which represents combined estrogenic contaminants' ecological risk, indicated no mentionable risk for predicted diluted effluents (HI = 0.0097 to 0.0218) as well as shoreline samples (HI = 0.393 to 0.522) in the receiving estuarine or marine waters.

A critical review on the distribution and ecological risk assessment of steroid hormones in the environment in China.

During past two decades, steroid hormones have raised significant public concerns due to their potential adverse effects on the hormonal functions of aquatic organisms and humans. Considering China being a big producer and consumer of steroid hormones, we summarize the current contamination status of steroid hormones in different environmental compartments in China, and preliminarily assess the associated risks to ecological systems. The results show that steroid hormones are ubiquitously present in Chinese surface waters where estrogens are the most studied steroids compared with androgens, progestogens and glucocorticoids. Estrone (E1), 17ß-estradiol (17ß-E2) and estriol (E3) are generally the dominant steroid estrogens in Chinese surface waters, whereas for the other steroids, androsterone (ADR), epi-androsterone (EADR), progesterone (PGT), cortisol (CRL) and cortisone (CRN) have relatively large contributions. Meanwhile, the investigations for the other environmental media such as particles, sediments, soils and groundwater have been limited, as well as for steroid conjugates and metabolites. The median risk quotients of most steroid hormones in surface waters and sediments are lower than 1, indicating low to moderate risks to local organisms. This review provides a full picture of steroid distribution and ecological risks in China, which may be useful for future monitoring and risk assessment. More studies may focus on the analysis of steroid conjugates, metabolites, solid phase fractions, analytical method development and acute/chronic toxicities in different matrices to pursue a more precise and holistic risk assessment.

Is an anodizing coating associated to the photobiomodulation able to optimize bone healing in ovariectomized animal model?

This in vivo study investigated whether the bioactivity of anodizing coating, produced by plasma electrolytic oxidation (PEO), on mini-plate in femur fracture could be improved with the association of photobiomodulation (PBM) therapy. From the 20 ovariectomized Wistar female rats, 8 were used for model characterization, and the remaining 12 were divided into four groups according to the use of PBM therapy by diode laser (808 nm; power: 100 mW; energy: 6.0 J; energy density: 212 J/cm2; power density: 3.5 W/cm2) and the type of mini-plate surface (commercially pure titanium mini-plate -cpTi- and PEO-treated mini-plate) as follow: cpTi; PEO; cpTi/PBM; and PEO/PBM. After 60 days of surgery, fracture healing underwent microstructural, bone turnover, histometric, and histologic adjacent muscle analysis. Animals of groups with PEO and PBM showed greater fracture healing than cpTi control group under histometric and microstructural analysis (P < 0.05); however, bone turnover was just improved in PBM's groups (P < 0.05). there was no difference between cpTi and PEO without PBM (P > 0.05). Adjacent muscle analysis showed no metallic particles or muscle alterations in all groups. PEO and PBM are effective strategies for bone repair in fractures, however their association does not provide additional advantages.

Study on the Release Potential of BPA and Steroid Estrogens in the Sediments of Erhai Lake, a Typical Plateau Lake of China.

On-site sampling analysis and laboratory-scale experiments were conducted to study the pollution status and release potential of EDCs in Erhai Lake. We found that nitrogen and phosphorus pollution in Erhai Lake sediment were both at a high level, as well as EDCs pollution. The concentrations of BPA, E2α, E1, E2ß, EE2, and E3 were 36.84 ng/g(DW), 13.04 ng/g(DW), 128.97 ng/g(DW), 52.57 ng/g(DW), 18.48 ng/g(DW) and 5.36 ng/g(DW), respectively. The concentrations of E2α, E1, E2ß and EE2 in the bottom water were higher than the surface water due to the impact of sediment release. The results of the 20 days release test indicated that BPA release from the sediment had a greater correlation with the original concentration and the particle size of sediment, while the steroid EDCs had no obvious correlation with these two factors, probably due to the difference in hydrophobicity between them. Under hydraulic disturbance and aerobic conditions, the release process of EDCs was accompanied by a large amount of microbial degradation, and degradation amount > released amount. BPA was released quickly, 9.56% was released in 20 days, but only 3.37% of steroid EDCs released. In comparison, the release process of steroids was longer and posed a greater threat to aquatic ecology.

Obesity-associated inflammation induces androgenic to estrogenic switch in the prostate gland.

BACKGROUND AND OBJECTIVE: Our patient cohort revealed that obesity is strongly associated with steroid-5α reductase type 2 (SRD5A2) promoter methylation and reduced protein expression. The underlying mechanism of prostatic growth in this population is poorly understood. Here we addressed the question of how obesity, inflammation, and steroid hormones affect the development of benign prostatic hyperplasia (BPH). MATERIAL AND METHODS: We used preadipocytes, macrophages, primary human prostatic stromal cells, prostate tissues from high-fat diet-induced obese mice, and 35 prostate specimens that were collected from patients who underwent transurethral resection of the prostate (TURP). RNA was isolated and quantified with RT-PCR. Genome DNA was extracted and SRD5A2 promoter methylation was determined. Sex hormones were determined by high-performance liquid chromatography-tandem mass spectrometry. Protein was extracted and determined by ELISA test. RESULTS: In prostatic tissues with obesity, the levels of inflammatory mediators were elevated. SRD5A2 promoter methylation was promoted, but SRD5A2 expression was inhibited. Inflammatory mediators and saturated fatty acid synergistically regulated aromatase activity. Obesity promoted an androgenic to estrogenic switch in the prostate. CONCLUSIONS: Our findings suggest that obesity-associated inflammation induces androgenic to estrogenic switch in the prostate gland, which may serve as an effective strategy for alternative therapies for management of lower urinary tract symptoms associated with BPH in select individuals.

[Effects of 17ß-E2 on growth and gonad development of overwintering Whitmania pigra].

The present study was conducted to explores the effects of short-term addition of 17ß-E2 on the growth,gonad development and internal quality of overwintering Whitmania pigra. Before overwintering,0. 0,1. 0,10. 0,25. 0,50. 0,100. 0 µg·L~(-1) of 17ß-E2 were added to the aquaculture water for 6 weeks and then hibernated for 60 days. The changes of growth performance,gonad index,morphological structure of spermary( ovary),endogenous steroid hormones level and internal quality were measured. The results showed that the body weight,weight gain rate,specific growth rate,female gonad index,oocyte development and endogenous estrogen level of the leech increased first and then decreased with the increase of the concentration of exogenous 17ß-E2,which were higher than those of the control group. The body weight,weight gain rate and specific growth rate of the leech at the concentration of 25 µg·L~(-1)17ß-E2 were significantly higher than those of the other groups( P<0. 05),oocyte development and endogenous estrogen levels were significantly higher than those of other groups at the concentration of 50 µg·L~(-1)( P<0. 05). When the concentration of exogenous 17ß-E2 was higher than 50 µg·L~(-1),the levels of male gonad index,spermatocyte development,endogenous androgen and progesterone were significantly inhibited( P< 0. 05). There was no significant difference in endogenous corticosteroid levels among the groups. In conclusion,short-term addition of exogenous 17ß-E2 of 10-25 µg·L~(-1) could promote the growth of overwintering leeches,oocyte development and antithrombin activity without inhibiting the development of male gonads.

Impact of Soil Filtration on Metals, Nutrients, and Estrogenic Activity of Reclaimed Water.

This study examined the impact of soil filtration on water quality for reclaimed water from two municipal wastewater treatment processes with two soils (forest and farm). Soils were watered for 16 wk. Nutrients (NO-N, NH-N, and PO), electrical conductivity, hardness, pH, total metals, and estrogenic activity in source and effluents were measured. Results were evaluated in reference to current regulations in Washington State. Both reclaimed waters met standards for unrestricted agricultural use but did not meet current state regulations for ecological use. Metal concentrations in reclaimed waters were generally similar to the control (tap water). Both source and soil filtered water met Washington State stream water criteria for chronic exposure for As but exceeded groundwater standards. Concentrations of Cd, Pb, Ag, and Zn in the effluents were similar or slightly elevated to reclaimed source water for both soils and water types. Soil filtration improved water quality to levels within Washington State regulatory limits for ecological use for Cd, Cu, and Zn but decreased water quality for Pb for both soil and reclaimed water types. Phosphate concentrations were reduced by soil filtration, but NO concentrations were increased for both soils and all water types above source water concentrations. The estrogenic activity of the sand-filtered water decreased to near detection limits in the forest soil and by >200% in the farm soil. The degradation in water quality (increased NO, As, and Mn) after soil filtration observed in this study would likely occur with any water type, including rainwater.

Screening Estrogen Receptor Modulators in a Paper-Based Breast Cancer Model.

The health risks associated with acute and prolonged exposure to estrogen receptor (ER) modulators has led to a concerted effort to identify and prioritize potential disruptors present in the environment. ER agonists and antagonists are identified with end-point assays, quantifying changes in cellular proliferation or gene transactivation in monolayers of estrogen receptor alpha expressing (ER+) cells upon exposure. While these monolayer cultures can be prepared, dosed, and analyzed in a highly parallelized manner, they are unable to predict the potencies of ER modulators in vivo accurately. Physiologically relevant model systems that better predict tissue- or organ-level responses are needed. To address this need, we describe here a screening platform capable of quantitatively assessing ER modulators in 96 chemically isolated 3D cultures. These cultures are supported in wax-patterned paper scaffolds whose design has improved performance and throughput over previously described paper-based setups. To highlight the potential of paper-based cultures for toxicity screens, we measured the potency of known ER modulators with a luciferase-based reporter assay. We also quantified the proliferation and invasion of two ER+ cell lines in the presence of estradiol. Despite the inability of the current setup to better predict in vivo potencies of ER modulators than monolayer cultures, the results demonstrate the potential of this platform to support increasingly complex and physiologically relevant tissue-like structures for environmental chemical risk assessment.

Fate of estrogens in a pilot-scale step-feed anoxic/oxic wastewater treatment system controlling by nitrogen and phosphorus removal.

The control measures for estrogens in the aquatic environment are topics of growing concern. It is a meaningful issue to finding optimal process parameters for efficient removal of estrogens with the purpose of efficient total nitrogen (TN) or total phosphorus (TP) removal in sewage treatment plants. The present paper is concerned with the relationships between the estrogen removal and TN or TP removal in a pilot-scale three-stage anoxic/oxic (A/O) system treating real municipal wastewater. The total removal efficiency for estrone (E1) and 17ß-estradiol (E2) and their sulfate and glucuronide conjugates were on average 87% in the pilot-scale system. The concentrations of the sulfate and glucuronide conjugates of estrogens (E1 and E2) in the system were much lower than the estrogens, which might be caused by the rapid degradation of conjugates in the pilot-scale system. The average removal efficiencies of E1 and E2 and their sulfate and glucuronide conjugates were significantly lower under high TP removal conditions than those under high TN removal conditions that suggested that the ammonia oxidation promotes estrogen degradation. When the system achieved efficient TN removal, the concentrations of both E1 and E2 were generally lower in the aerobic zones than those in the anoxic zones. Instead, when the system achieved efficient TP removal conditions, the estrogen concentrations were higher in the aerobic zones than in the anoxic zones. However, it was thought that the variation of the concentrations of the estrogen conjugates had weak influence on concentrations of the free estrogens. The increase of the free estrogens in the aerobic zones could be attributed to the release of the estrogens adsorbed on the sludge. The variation of estrogens in a three-stage A/O system can be properly estimated and measured by a binary linear regression model with the variables of TP and TON (NO2--N and NO3--N), which is probably the important information for the improvement and optimization of wastewater treatment processes to obtain higher removal efficiency for estrogens.

Estrogenic Activities of Food Supplements and Beers as Assessed by a Yeast Bioreporter Assay.

Mounting evidence of the effects of endocrine-disrupting chemicals (EDCs) in humans has led to assaying a vast array of food items (processed or packaged) as possible sources of human exposure to estrogens. In this study, we investigated the current situation in this respect of different food supplements and beer brands. Eleven food supplements and 24 beer brands were obtained from Helsinki, Finland. Sample preparation was carried out by established methods while estrogenic activities were assessed by a yeast bioluminescent assay, using two recombinant yeast strains (Saccharomyces cerevisiae BMAEREluc/ERα and S. cerevisiae BMA64/luc). All the food supplements as well as 81% of the beer samples tested were found to be estrogenic, with estradiol equivalent concentrations of food supplements and beer brands ranging from 7.5 to 11.5 µg/ml and from below detection limits to 43.6 ng/ml, respectively. The estrogenic activities detected in beer samples were not dependent on the beer's alcoholic content, the country of production, or the size of the production brewery. The results of our study imply that both food supplements and beers can be a significant source of human exposure to estrogens. Therefore, further studies and regular surveillance are warranted.

Steroids in traditional Chinese medicine: what is the evidence?

Local healthcare providers often question the possible steroidal activity of traditional Chinese medicine (TCM) herbs or herbal products and implicate them as a cause for adrenal insufficiency or Cushing's syndrome in patients with a history of TCM intake. We conducted a comprehensive database search for evidence of potential glucocorticoid, mineralocorticoid, androgenic or oestrogenic activity of herbs or herbal products. Overall, there are not many herbs whose steroidal activity is well established; among these, most cases were based on preclinical studies. Liquorice root may cause pseudoaldosteronism through interference with the steroidogenesis pathway. Although ginseng and cordyceps have some in vitro glucocorticoid activities, the corroborating clinical data is lacking. Deer musk and deer antler contain androgenic steroids, while epimedium has oestrogenic activity. On the other hand, adulteration of herbal products with exogenous glucocorticoids is a recurrent problem encountered locally in illegal products masquerading as TCM. Healthcare providers should stay vigilant and report any suspicion to the relevant authorities for further investigations.

Multiresidue determination of estrogens in different dairy products by ultra-high-performance liquid chromatography triple quadrupole mass spectrometry.

In this work, a simple and fast methodology has been validated and applied for the analysis of a group of 22 estrogenic compounds including eight phytoestrogens (i.e. daidzein, enterodiol, glycitein, enterolactone, genistein, formononetin, prunetin, biochanin A), six mycotoxins (ß-zearalanol, ß-zearalenol, α-zearalanol, α-zearalenol, zearalanone, zearalenone) as well as four synthetic (i.e. ethynylestradiol, diethylstilbestrol, dienestrol, hexestrol) and four natural estrogens (i.e. estriol, 17ß-estradiol, 17α-estradiol, estrone) in different dairy products. Extraction was carried out using the QuEChERS method while separation, determination and quantification of the target analytes were achieved by ultra-high-performance liquid chromatography coupled to triple quadrupole mass spectrometry with an electrospray ionization interface. The methodology was validated for four dairy product samples with relevant interest for the population including skimmed and whole cheese and goat and cow kefir, using 17ß-estradiol-2,4,16,16,17-d5 as internal standard for natural and synthetic estrogens and ß-zeralanol-10,10,11,12,12-d5 as internal standard for mycotoxins and phytoestrogens. Recovery ranged from 70 to 119% for the four types of matrices with RSD values lower than 14% and the limits of quantification of the method achieved were in the range 0.025-2.50µg/kg for all samples. Finally, the analysis of commercially available products was carried out finding the presence of daidzein, glycitein enterolactone and genistein in some of the studied samples.

Phytoestrogens and sterols in waters with cyanobacterial blooms - Analytical methods and estrogenic potencies.

Compounds with estrogenic potencies and their adverse effects in surface waters have received much attention. Both anthropogenic and natural compounds contribute to overall estrogenic activity in freshwaters. Recently, estrogenic potencies were also found to be associated with cyanobacteria and their blooms in surface waters. The present study developed and compared the solid phase extraction and LC-MS/MS analytical approaches for determination of phytoestrogens (8 flavonoids - biochanin A, coumestrol, daidzein, equol, formononetin, genistein, naringenin, apigenin - and 5 sterols - ergosterol, ß-sitosterol, stigmasterol, campesterol, brassicasterol) and cholesterol in water. The method was used for analyses of samples collected in stagnant water bodies dominated by different cyanobacterial species. Concentrations of individual flavonoids ranged from below the limit of detection to 3.58 ng/L. Sterols were present in higher amounts up to 2.25 µg/L. Biological potencies of these phytoestrogens in vitro were characterized using the hERα-HeLa-9903 cell line. The relative estrogenic potencies (compared to model estrogen - 17ß-estradiol) of flavonoids ranged from 2.25E-05 to 1.26E-03 with coumestrol being the most potent. None of the sterols elicited estrogenic response in the used bioassay. Estrogenic activity was detected in collected field water samples (maximum effect corresponding to 2.07 ng/L of 17ß-estradiol equivalents, transcriptional assay). At maximum phytoestrogens accounted for only 1.56 pg/L of 17ß-estradiol equivalents, contributing maximally 8.5% of the total estrogenicity of the water samples. Other compounds therefore, most likely of anthropogenic origin such as steroid estrogens, are probably the major drivers of total estrogenic effects in these surface waters.

Estrogen Transport in Surface Runoff from Agricultural Fields Treated with Two Application Methods of Dairy Manure.

This study compares two methods of dairy manure application-surface broadcast and shallow disk injection-on the fate and transport of natural estrogens in surface runoff from 12 field plots in central Pennsylvania. Ten natural surface runoff events were sampled over a 9-mo period after fall manure application. Results show that the range of estrogen concentrations observed in surface runoff from the broadcast plots was several orders of magnitude higher (>5000 ng L) than the concentrations in runoff from the shallow disk injection plots (<10 ng L). Additionally, the transport dynamics differed, with the majority of the estrogen loads from the surface broadcast plots occurring during the first rainfall event after application, whereas the majority of the loads from the shallow disk injection plots occurred more than 6 mo later during a hail storm event. Total estrogen loads were, on average, two orders of magnitude lower for shallow disk injection compared with surface broadcast. Independent of the method of manure application, 17α-estradiol and estrone were preserved in the field for as long as 9 mo after application. Overall, injection of manure shows promise in reducing the potential for off-site losses of hormones from manure-amended soils.

Estrogenic compounds in Tunisian urban sewage treatment plant: occurrence, removal and ecotoxicological impact of sewage discharge and sludge disposal.

The occurrence, fate and ecotoxicological assessment of selected estrogenic compounds were investigated at Tunisian urban sewage treatment plant. The influents, effluents, as well as primary, secondary and dehydrated sludge, were sampled and analyzed for the target estrogens to evaluate their fate. All target compounds were detected in both sewage and sludge with mean concentrations from 0.062 to 0.993 µg L-1 and from 11.8 to 792.9 µg kg-1dry weight, respectively. A wide range of removal efficiencies during the treatment processes were observed, from 6.3 % for estrone to 76.8 % for estriol. Ecotoxicological risk assessment revealed that the highest ecotoxicological risk in sewage effluent and dehydrated sludge was due to 17ß-estradiol with a risk quotient (RQ) of 4.6 and 181.9, respectively, and 17α-ethinylestradiol with RQ of 9.8 and 14.85, respectively. Ecotoxicological risk after sewage discharge and sludge disposal was limited to the presence of 17ß-estradiol in dehydrated-sludge amended soil with RQ of 1.38. Further control of estrogenic hormones in sewage effluent and sludge is essential before their discharge and application in order to prevent their introduction into the natural environment.