Endocrine disruptors in bottled mineral water: total estrogenic burden and migration from plastic bottles

Background, aim, and scope  Food consumption is an important route of human exposure to endocrine-disrupting chemicals. So far, this has been demonstrated by exposure modeling or analytical identification of single substances in foodstuff (e.g., phthalates) and human body fluids (e.g., urine and blood). Since the research in this field is focused on few chemicals (and thus missing mixture effects), the overall contamination of edibles with xenohormones is largely unknown. The aim of this study was to assess the integrated estrogenic burden of bottled mineral water as model foodstuff and to characterize the potential sources of the estrogenic contamination. Materials, methods, and results  In the present study, we analyzed commercially available mineral water in an in vitro system with the human estrogen receptor alpha and detected estrogenic contamination in 60% of all samples with a maximum activity equivalent to 75.2 ng/l of the natural sex hormone 17β-estradiol. Furthermore, breeding of the molluskan model Potamopyrgus antipodarum in water bottles made of glass and plastic [polyethylene terephthalate (PET)] resulted in an increased reproductive output of snails cultured in PET bottles. This provides first evidence that substances leaching from plastic food packaging materials act as functional estrogens in vivo. Discussion and conclusions  Our results demonstrate a widespread contamination of mineral water with xenoestrogens that partly originates from compounds leaching from the plastic packaging material. These substances possess potent estrogenic activity in vivo in a molluskan sentinel. Overall, the results indicate that a broader range of foodstuff may be contaminated with endocrine disruptors when packed in plastics.     

基于MIKE11模型的南淝河及其支流内分泌干扰风险评价与调控

为寻求评价和调控城市河流内分泌干扰风险的有效方法,以城市河流典型代表―南淝河及其支流水系为例,运用MIKE11水动力学和对流扩散模块构建南淝河流域城市河流雌激素迁移转化模型.同时,利用该模型对南淝河水系雌激素类物质引起的内分泌干扰风险进行评价,并基于未来可达水污染控制目标提出风险控制措施.结果表明,在最优雌激素排放率情景下,约50%以上的河段具有潜在内分泌干扰风险;仅在最高排放率情景下,丰、平、枯水期分别有19%、53%和67%河段存在高风险,高风险河段集中在南淝河城区段及其支流二十埠河、板桥河和店埠河.3种削减内分泌干扰风险远近期可达情境方案分析表明:仅依靠单一的点源截污控制方案,即使在最优排放率情景下仍有近50%的河段具有潜在风险水平;通过点源截污和70%的面源排污控制相结合的调控措施,在最优排放率情景下能使所有河段处于无风险水平;在将点源和面源排污完全截留控制的远期目标下,可以基本实现预期情景下90%以上河段处于无内分泌干扰风险水平.     

环境对阿尔茨海默病病因影响

阿尔茨海默病是一种发生于老年人群的原发生退行性脑病。目前，关于ＡＤ病研究中的中心问题是弄清楚其发病的病理，如果可以抑制的一步和几步，将减缓或防止其发病，因此搞清ＡＤ病的发病机理对预防和治疗ＡＤ病相当重要。     

蛋白质组学技术在环境内分泌干扰物分析上应用

为了探讨蛋白质组学技术在环境雌激素分析上的应用,用浓度为3.7×10-9mol/L的17β-雌二醇(E2)处理MCF-7细胞2或3天后进行双向电泳,对比不同的蛋白质提取、染色方法,利用光密度扫描仪将电泳图像数字化,并使用PDQuest软件进行图像分析,以观察雌激素诱导下的蛋白质表达图谱。结果表明,在蛋白质的提取上,冻融法比超声裂解法更有效;在蛋白质染色上,银染法比考马斯亮蓝染色法能得到更多可显示的蛋白质斑点。用E2处理72h后的MCF-7细胞,可检测到40个下调蛋白和5个上调蛋白。该蛋白质表达模式的改变提示了环境雌激素诱导下的特异性蛋白质表达图谱变化。本研究为分析雌激素类内分泌干扰物的内在作用机制提供了一种新的可行方法,也为建立其高通量筛选方法提供了新的思路。     

双酚A体外类雌激素活性评价及其作用机制的初步研究

采用体外MCF-7细胞增殖试验检测了双酚A(BPA)的类雌激素活性，并用生长曲线分析、细胞周期分析、他奠昔芬拮抗试验和凋亡检测对其作用机制进行了初步探讨。结果显示BPA有明显刺激MCF-7细胞增殖的类雌激素活性；细胞周期分析BPA组细胞增殖指数均明显高于溶剂对照组；BPA刺激MCF-7细胞增殖的类雌激素活性可被Tam拮抗：BPA能明显抑制MCF-7细胞凋亡的发生。结果提示BPA刺激MCF-7细胞增殖的类雌激素活性作用机制可能是BPA与雌激素受体结合后，影响细胞增殖及细胞凋亡过程，从而产生一系列生物效应。     

Ecotoxicological Assessment of Sediment, Suspended Matter and Water Samples in the Upper Danube River. A pilot study in search for the causes for the decline of fish catches (12 pp)

Goals, Scope and Background Fish populations, especially those of the grayling (Thymallus thymallus), have declined over the last two decades in the upper Danube River between Sigmaringen and Ulm, despite intensive and continuous stocking and improvement of water quality since the 1970s. Similar problems have been reported for other rivers, e.g. in Switzerland, Great Britain, the United States and Canada. In order to assess if ecotoxicological effects might be related to the decline in fish catch at the upper Danube River, sediment, suspended matter and waste water samples from sewage treatment plants were collected at selected locations and analyzed in a bioanalytical approach using a battery of bioassays. The results of this pilot study will be used to decide if a comprehensive weight-of-evidence study is needed. Methods Freeze-dried sediments and suspended particulate matters were extracted with acetone in a Soxhlet apparatus. Organic pollutants from sewage water were concentrated using XAD-resins. In order to investigate the ecotoxicological burden, the following bioassays were used: (1) neutral red assay with RTL-W1 cells (cytotoxicity), (2) comet assay with RTLW1 cells (genotoxicity), (3) Arthrobacter globiformis dehydrogenase assay (toxicity to bacteria), (4) yeast estrogen screen assay (endocrine disruption), (5) fish egg assay with the zebrafish (Danio rerio; embryo toxicity) and (6) Ames test with TA98 (mutagenicity). Results and Discussion The results of the in vitro tests elucidated a considerable genotoxic, cytotoxic, mutagenic, bacteriotoxic, embryotoxic and estrogenic burden in the upper Danube River, although with a very inhomogeneous distribution of effects. The samples taken from Riedlingen, for example, induced low embryo toxicity, but the second highest 17β-estradiol equivalent concentration (1.8 ng/L). Using the fish egg assay with native sediments, a broad range of embryotoxic effects could be elucidated, with clear-cut dose-response relationships for the embryotoxic effects of contaminated sediments. With native sediments, embryotoxicity was clearly higher than with corresponding pore waters, thus corroborating the view that – at least for fish eggs – the bioavailability of particle-bound lipophilic substances in native sediments is higher than generally assumed. The effect observed most frequently in the fish egg assay was a developmental delay. A comparison of our own results with locations along the rivers Rhine and Neckar demonstrated similar or even higher ranges of ecotoxicological burdens in the Danube River. Conclusions The complex pattern of ecotoxicological effects caused by environmental samples from the Danube River, when assessed in an in vitro biotest battery using both acute and more specific endpoints, showed that integration of different endpoints is essential for appropriate hazard assessment. Overall, the ecotoxicological hazard potential shown has indeed to be considered as one potential reason for the decline in fish catches at the upper Danube River. However, based on the results of this pilot study, it is not possible to elucidate that chemically induced alterations are responsible for the fish decline. Recommendations and Perspective . In order to confirm the ecological relevance of the in vitro results for the situation in the field and especially for the decline of the grayling and other fishes, further integrated investigations are required. For linking the weight of evidence obtained by in vitro assays and fish population investigations, the application of additional, more specific biomarkers (e.g. vitellogenin induction, EROD and micronucleus assay) has been initiated in fish taken from the field as well as in situ investigations.     

ERα和ERβ介导的有机磷农药雌激素效应差异分析

雌激素受体（Estrogen Receptor,ER）ERα和ERβ能与雌激素特异性结合,它们在结构上的差异性可能影响与类雌激素的相互作用,这进而会影响其功能.因此,针对10种有机磷农药对ERα和ERβ的雌激素效应差异,采用分子对接方法比较分析了它们与ERα、ERβ相互作用的差异并筛选出与ERα和ERβ相互作用的最佳结合模式.结果发现,10种有机磷农药分子与ERα配体结合域（Ligand Binding Domain,LBD）精氨酸（Arg394）、赖氨酸（Lys449）、色氨酸（Trp393）形成氢键;而部分农药分子与ERβLBD精氨酸（Arg346）、亮氨酸（Leu343）、赖氨酸（Lys401）形成氢键.有机磷农药配体与ERαLBDArg394、Lys449、苯丙氨酸（Phe445）、脯氨酸（Pro324）、Leu387、谷氨酸（Glu353）、甘氨酸（Gly390）等存在疏水作用,与ERβLBD氨基酸Arg346、Glu305、Leu343、蛋氨酸（Met336）、组氨酸（His279）、丙氨酸（Ala302）等存在疏水作用.采用量子化学从头算进一步分析了10种有机磷农药分子表面静电势的差异.研究发现,有机磷农药分子与ERα和ERβ受体不同的相互作用是导致雌激素效应差异的重要原因.     

江苏某地长江、太湖水源水及出厂水中有机物雌激素活性的比较

于2010年10月～2012年3月期间4次采集江苏WX和SZ地区长江、太湖水源水及相关自来水厂出厂水共43个水样,通过XAD-2树脂吸附、洗脱、浓缩其中的非挥发性有机物(nonvolatile organic compounds,NVOCs);应用重组酵母雌激素筛检试验(recombinant yeast estrogen screen,YES试验)检测和比较水中有机物雌激素活性水平.结果表明所有长江、太湖水源水样均具有雌激素活性,太湖水源水的雌二醇当量(EEQs)为0.04～2.07 ng.L-1,EC25-E2水样量为5.78～207.98 mL,长江水源水的雌二醇当量(EEQs)为0.69～1.15 ng.L-1,EC25-E2水样量为9.06～31.62 mL;SZ地区太湖水源水中具类雌激素内分泌干扰物的污染程度(0.69 ng.L-1)高于WX地区(0.44 ng.L-1);长江、太湖水源水经自来水厂常规净水工艺处理后,58.3%的出厂水样未检出雌激素活性,长江水源自来水厂可降低雌激素活性80.3%～100%,而太湖水源自来水厂可降低55.5%～100%.     

Removal of estrogens by electrochemical oxidation process

Treatments of estrogens such as Estrone(E1), Estradiol(E2) and Ethinylestradiol(EE2) were conducted using an electrolytic reactor equipped with multi-packed granular glassy carbon electrodes.Experimental results showed that E1, E2 and EE2 were oxidized in the range of 0.45–0.85 V and were removed through electro-polymerization. Observed data from continuous experiments were in good agreement with calculated results by a mathematical model constructed based on mass transfer limitation. In continuous treatment of trace estrogens(1 μg/L), 98% of E1, E2 and EE2 were stably removed. At high loading rate(100 μg/L), removal efficiency of E1 was kept around 74%–88% for21 days, but removal efficiency reduced due to passivation of electrodes. However, removal efficiency was recovered after electrochemical regeneration of electrodes in presence of ozone. Electric energy consumption was observed in the range of 1–2 Wh/m3. From these results, we concluded that the present electrochemical process would be an alternative removal of estrogens.     

有机氯类农药在残留剂量下联合诱导乳腺癌MCF-7细胞增殖的机制研究

有机氯类农药为已确定的环境雌激素,易在体内蓄积产生慢性毒性危害人体健康。选择单一存在时无明显雌激素效应的残留剂量进行联合试验,观察联合作用后雌激素效应的增减。选取乳腺癌MCF-7细胞作为研究对象,对残留剂量下六氯苯(HCB)、β-六六六(β-BHC)和p,p’-滴滴涕(p,p’-DDT)联合后的雌激素效应机制进行了研究。首先,运用MTT法观察3种有机氯类农药残留剂量下单独和联合作用对MCF-7细胞生长的影响;然后,运用流式细胞仪测定有机氯类农药联合对MCF-7细胞周期的影响;之后,采用Western Blot法测定有机氯类农药联合后ERα、ERβ、ERK1/2、p-ERK1/2、Ki67、c-Myc和Cyclin D1蛋白表达的变化。结果显示,β-BHC+p,p’-DDT组、HCB+β-BHC+p,p’-DDT组作用24 h后细胞增殖率为115.0%、120.1%,作用48 h后增殖率为121.2%、126.3%,其他联合组与对照相比未发生明显变化;β-BHC+p,p’-DDT组、HCB+β-BHC+p,p’-DDT组中MCF-7细胞的G1期细胞比例下降,S期细胞比例上升,细胞呈高增殖状态;β-BHC+p,p’-DDT组、HCB+β-BHC+p,p’-DDT组和雌二醇(E2)组作用48 h后,均降低ERα的蛋白表达,升高ERβ的蛋白表达,促进p-ERK1/2、Ki67、Cyclin D1和c-Myc的蛋白表达。研究表明,在单一作用时不具有雌激素效应的残留剂量下,仅β-BHC+p,p’-DDT联合组和HCB+β-BHC+p,p’-DDT联合组具有明显雌激素效应,其效应机制为经雌激素受体途径,激活ERβ蛋白表达,抑制ERα蛋白表达,促进p-ERK1/2、Ki67、c-Myc和Cyclin D1等相关蛋白的表达。