The fate of steroid estrogens: Partitioning during wastewater treatment and onto river sediments

This is the author's accepted manuscript. The final published article is available from the link below. Copyright @ 2010 Springer Science+Business Media B.V.The partitioning of steroid estrogens in wastewater treatment and receiving waters is likely to influence their discharge to, and persistence in, the environment. This study investigated the partitioning behaviour of steroid estrogens in both laboratory and field studies. Partitioning onto activated sludge from laboratory-scale Husmann units was rapid with equilibrium achieved after 1 h. Sorption isotherms and Kd values decreased in the order 17α-ethinyl estradiol > 17α-estradiol > estrone > estriol without a sorption limit being achieved (1/n >1). Samples from a wastewater treatment works indicated no accumulation of steroid estrogens in solids from primary or secondary biological treatment, however, a range of steroid estrogens were identified in sediment samples from the River Thames. This would indicate that partitioning in the environment may play a role in the long-term fate of estrogens, with an indication that they will be recalcitrant in anaerobic conditions.EPSR

A new method for the separation of androgens from estrogens and for the partition of estriol from the estrone-estradiol fraction: with special reference to the identification and quantitative microdetermination of estrogens by ultraviolet absorption spectrophotometry

It is recognized generally that a qualitative and quantitative knowledge of the excretion pattern of the urinary estrogens is one index to an understanding of the functional activity of the ovary and adrenal cortex. Obviously, such determinations may be useful also in evaluating the normal and abnormal functions of other physiologically related endocrine glands as well as of organs like the liver and kidneys. The clinical applications of these data are self-evident. Various attempts have been made to circumvent the notoriously inaccurate values which have been obtained for the urinary estrogens by a variety of bioassay methods and calorimetric techniques (1, 2). The acknowledged shortcomings of these methods have led us to investigate the application of ultraviolet absorption spectrophotometry to the quantitative determination of the urinary estrogens in an attempt to develop an objective physical method for their accurate determination. It is known that the infra-red portion of the spectrum yields more differentially characteristic curves, but those of the ultraviolet range are more readily obtainable, and consequently better adapted to clinical use. This communication is concerned with studies of the following aspects of the problem: (1) spectrophotometric identification and quantitative micro determination of crystalline estrogens; (2) detection by spectrophotometric assay of gross errors in current methods for extraction and partition of estrogens; (3) studies on the ultraviolet absorption of substances comprising the background material; (4) separation of the phenolic estrogens from the so called neutral steroid fraction; (5) separation of urinary estrogens from other urinary phenolic substances by steam distillation; (6) micro-Girard separation of estrone from estradiol; (7) an essentially new method for the extraction and partition of crystalline estrone, estradiol, and estriol, and their quantitative assay by ultraviolet spectrophotometry

Catechol estrogens stimulate insulin secretion in pancreatic β-cells via activation of the transient receptor potential A1 (TRPA1) channel

Estrogen hormones play an important role in controlling glucose homeostasis and pancreatic β-cell function. Despite the significance of estrogen hormones for regulation of glucose metabolism, little is known about the roles of endogenous estrogen metabolites in modulating pancreatic β-cell function. In this study, we evaluated the effects of major natural estrogen metabolites, catechol estrogens, on insulin secretion in pancreatic β-cells. We show that catechol estrogens, hydroxylated at positions C2 and C4 of the steroid A ring, rapidly potentiated glucose-induced insulin secretion via a nongenomic mechanism. 2-Hydroxyestrone, the most abundant endogenous estrogen metabolite, was more efficacious in stimulating insulin secretion than any other tested catechol estrogens. In insulin-secreting cells, catechol estrogens produced rapid activation of calcium influx and elevation in cytosolic free calcium. Catechol estrogens also generated sustained elevations in cytosolic free calcium and evoked inward ion current in HEK293 cells expressing the transient receptor potential A1 (TRPA1) cation channel. Calcium influx and insulin secretion stimulated by estrogen metabolites were dependent on the TRPA1 activity and inhibited with the channel-specific pharmacological antagonists or the siRNA. Our results suggest the role of estrogen metabolism in a direct regulation of TRPA1 activity with potential implications for metabolic diseases

Estrogens promote misfolded proinsulin degradation to protect insulin production and delay diabetes

Summary: Conjugated estrogens (CE) delay the onset of type 2 diabetes (T2D) in postmenopausal women, but the mechanism is unclear. In T2D, the endoplasmic reticulum (ER) fails to promote proinsulin folding and, in failing to do so, promotes ER stress and β cell dysfunction. We show that CE prevent insulin-deficient diabetes in male and in female Akita mice using a model of misfolded proinsulin. CE stabilize the ER-associated protein degradation (ERAD) system and promote misfolded proinsulin proteasomal degradation. This involves activation of nuclear and membrane estrogen receptor-α (ERα), promoting transcriptional repression and proteasomal degradation of the ubiquitin-conjugating enzyme and ERAD degrader, UBC6e. The selective ERα modulator bazedoxifene mimics CE protection of β cells in females but not in males. : Estrogens prevent diabetes in women, but the mechanism is poorly understood. Xu et al. report that estrogens activate the endoplasmic-reticulum-associated protein degradation pathway, which promotes misfolded proinsulin degradation, suppresses endoplasmic reticulum stress, and protects insulin secretion in mice and in human pancreatic β cells. Keywords: estrogens, beta cell, islet, endoplasmic reticulum stress, proinsulin misfolding, diabetes, bazedoxifene, sex dimorphism, ERAD, SER

Influence of operating parameters on the biodegradation of steroid estrogens and nonylphenolic compounds during biological wastewater treatment processes

This document is the unedited author's version of a Submitted Work that was subsequently accepted for publication in Environmental Science & Technology, copyright © American Chemical Society after peer review. To access the final edited and published work see http://pubs.acs.org/doi/abs/10.1021/es901612v.This study investigated operational factors influencing the removal of steroid estrogens and nonylphenolic compounds in two sewage treatment works, one a nitrifying/denitrifying activated sludge plant and the other a nitrifying/denitrifying activated sludge plant with phosphorus removal. Removal efficiencies of >90% for steroid estrogens and for longer chain nonylphenol ethoxylates (NP4−12EO) were observed at both works, which had equal sludge ages of 13 days. However, the biological activity in terms of milligrams of estrogen removed per day per tonne of biomass was found to be 50−60% more efficient in the nitrifying/denitrifying activated sludge works compared to the works which additionally incorporated phosphorus removal. A temperature reduction of 6 °C had no impact on the removal of free estrogens, but removal of the conjugated estrone-3-sulfate was reduced by 20%. The apparent biomass sorption (LogKp) values were greater in the nitrifying/denitrifying works than those in the nitrifying/denitrifying works with phosphorus removal for both steroid estrogens and nonylphenolic compounds possibly indicating a different cell surface structure and therefore microbial population. The difference in biological activity (mg tonne−1 d−1) identified in this study, of up to seven times, suggests that there is the potential for enhancing the removal of estrogens and nonylphenols if more detailed knowledge of the factors responsible for these differences can be identified and maximized, thus potentially improving the quality of receiving waters.Public Utilities Board (Singapore), Anglian Water Ltd, Severn Trent Water Ltd, Thames Water Utilities Ltd, United Utilities 393 Plc and Yorkshire Water Services

Biotransformation and bioconcentration of steroid estrogens by Chlorella vulgaris

The biotransformation and bioconcentration of natural and synthetic steroid estrogens by Chlorella vulgaris were investigated using batch shaking experiments with incubation for 48 hours in the light or dark. Estradiol and estrone were inter-convertible in both light and dark conditions, however this biotransformation showed a preference to estrone. In the light, 50% of estradiol was further metabolized to an unknown product. Apart from biotransformation, estrone as well as hydroxyestrone, estriol and ethinylestradiol were relatively stable in the algal culture, while estradiol valerate was hydrolyzed to estradiol and then estrone within 3 hours of incubation. All the tested estrogens exhibited a degree of partitioning to C. vulgaris, however, the concentrations of estriol, hydroxyestrone, ethinylestradiol and estradiol valerate were always below the quantification limits. For estradiol and estrone, the partitioning of these estrogens in the algal extracts to the filtrates was below 6% of the total present. The average concentration factor for estrone was around 27, however the concentration factor for estradiol is not reported since no equilibrium was reached between aqueous solution and that within the cells due to continuing biotransformation

The significance of sample mass in the analysis of steroid estrogens in sewage sludges and the derivation of partition coefficients in wastewaters

Optimization of an analytical method for determination of steroid estrogens, through minimizing sample size, resulted in recoveries >84%, with relative standard deviations <3% and demonstrated the significance of sample size on method performance. Limits of detection were 2.1–5.3 ng/g. Primary sludges had estrogen concentrations of up to one order of magnitude less than those found in biological sludges (up to 994 ng/g). However, partition coefficients were higher in primary sludges (except estriol), with the most hydrophobic compound (ethinylestradiol) exhibiting the highest Kp value, information which may be of value to those involved in modeling removal during wastewater treatment

Treatment and removal strategies for estrogens from wastewater

Natural and synthetic steroidal estrogens (estrone, 17β-estradiol and 17α-ethinylestradiol) are endocrine disrupters, that are discharged consistently from the sewage treatment works into surface waters, thereby causing endocrine disrupting effects to aquatic organisms at trace concentrations (nanogram per litre). Several years of research have been focused on their fate, behaviour and removal in the environment but primarily in the sewage treatment works which acts as a sink for these compounds. This review attempts to summarize the factors involved in the removal of these chemicals from the sewage treatment works. Biological processes, and to a limited extent physio-chemical properties, play a vital role in the endocrinal deactivation of which these compounds. The efficiency of these processes is highly dependent on operating parameters (such as sludge retention time, redox potential, etc) that govern the secondary treatment process of a functional sewage treatment works. Although advanced treatment technologies are available, cost and operational considerations do not make them a sustainable solution

The effectiveness of anaerobic digestion in removing estrogens and nonylphenol ethoxylates

This is the post-print version of the final paper published in Journal of Hazardous Materials. The published article is available from the link below. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. Copyright @ 2011 Elsevier B.V.The fate and behaviour of two groups of endocrine disrupting chemicals, steroid estrogens and nonylphenol ethoxylates, have been evaluated during the anaerobic digestion of primary and mixed sewage sludge under mesophilic and thermophilic conditions. Digestion occurred over six retention times, in laboratory scale reactors, treating sludges collected from a sewage treatment works in the United Kingdom. It has been established that sludge concentrations of both groups of compounds demonstrated temporal variations and that concentrations in mixed sludge were influenced by the presence of waste activated sludge as a result of transformations during aerobic treatment. The biodegradation of total steroid estrogens was >50% during primary sludge digestion with lower removals observed for mixed sludge, which reflected bulk organic solids removal efficiencies. The removal of nonylphenol ethoxylates was greater in mixed sludge digestion (>58%) compared with primary sludge digestion and did not reflect bulk organic removal efficiencies. It is apparent that anaerobic digestion reduces the concentrations of these compounds, and would therefore be expected to confer a degree of protection against exposure and transfer of both groups of compounds to the receiving/re-use environment.Thames Water, Yorkshire Water, and EPSRC

Disruption of 3D MCF-12A breast cell cultures by estrogens - An in vitro model for ER-mediated changes indicative of hormonal carcinogenesis

Copyright @ 2012 The Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and 85 reproduction in any medium, provided the original author and source are credited. The article was made available through the Brunel University Open Access Publishing Fund.Introduction: Estrogens regulate the proliferation of normal and neoplastic breast epithelium. Although the intracellular mechanisms of estrogens in the breast are largely understood, little is known about how they induce changes in the structure of the mammary epithelium, which are characteristic of breast cancer. In vitro three dimensional (3D) cultures of immortalised breast epithelial cells recapitulate features of the breast epithelium in vivo, including formation of growth arrested acini with hollow lumen and basement membrane. This model can also reproduce features of malignant transformation and breast cancer, such as increased cellular proliferation and filling of the lumen. However, a system where a connection between estrogen receptor (ER) activation and disruption of acini formation can be studied to elucidate the role of estrogens is still missing. Methods/Principal Findings: We describe an in vitro 3D model for breast glandular structure development, using breast epithelial MCF-12A cells cultured in a reconstituted basement membrane matrix. These cells are estrogen receptor (ER)α, ERβ and G-protein coupled estrogen receptor 1 (GPER) competent, allowing the investigation of the effects of estrogens on mammary gland formation and disruption. Under normal conditions, MCF-12A cells formed organised acini, with deposition of basement membrane and hollow lumen. However, treatment with 17β-estradiol, and the exogenous estrogens bisphenol A and propylparaben resulted in deformed acini and filling of the acinar lumen. When these chemicals were combined with ER and GPER inhibitors (ICI 182,780 and G-15, respectively), the deformed acini recovered normal features, such as a spheroid shape, proliferative arrest and luminal clearing, suggesting a role for the ER and GPER in the estrogenic disruption of acinar formation. Conclusion: This new model offers the opportunity to better understand the role of the ER and GPER in the morphogenesis of breast glandular structure as well as the events implicated in breast cancer initiation and progression.This study was partly funded by a School of Pharmacy Studentship