3,3',5-Triiodothyroacetic acid (TRIAC) was identified as a major contributor to the activity of thyroid hormone receptor (TR) agonists in environmental water. TRIAC contributed 60-148% of the TR-agonist activity in effluents from sewage treatment plants (STPs). Meanwhile, the contributions of 3,5,3'-triiodothyronine (T3), 3,3',5,5'-tetraiodothyronine (T4), and analogues were <1%. TRIAC concentrations in the range of 0.30-4.2 ng/L are likely enough to cause disruption of the thyroid system in living aquatic organisms. The origin of TRIAC in the STP effluents was investigated by analyzing both STP influents and effluents. Relatively high concentrations of T3 and T4 (2.5 and 6.3 ng/L, respectively) were found only in the influents. TRIAC was identified only in the effluents. These findings suggested that T3 and T4 in STP influents were potentially converted into TRIAC during activated sludge treatment or by other means. The evaluation of TRIAC at relevant environmental concentrations by in vivo assays and an appropriate treatment to reduce the TR activity in sewage are needed.

The human thyroid receptor (hTR)-agonist activities of 796 compounds were evaluated using a yeast two-hybrid assay in this study. A total of 17 compounds exhibited agonist activity at tenfold the effective concentration (EC×10) [0.0083–7,500 nM]. Additionally, TRIAC, TETRAC, and GC-1, which exhibited 30–200 times higher activities than T3 and T4, may cause thyroid-hormone receptors (TR) activity at extremely low concentrations in real environmental water. Moreover, 3-chloro-3',5,5'-triiodo-L-thyronine exhibited high TR-agonist activity for the first time. Chemicals with a structure equivalent to thyroid hormones exhibited TR-agonist activity regardless of the TR type. Information on hTR-agonist activity is important because previous studies, such as those involving EDSP21, have focused exclusively on rTR-agonist activity. Therefore, the knowledge gained from the present study will boost chemical regulation strategies benefiting human and wildlife.

The human thyroid receptor (hTR)-antagonist activities of 691 compounds were evaluated using a yeast two-hybrid assay with Saccharomyces cerevisiae Y190 introduced hTR alpha and coactivator. In parallel, those YTOX tests were conducted to evaluate whether those compounds affected either antagonism or toxicity. This is the first report that focuses on the hTR-antagonist activity of many chemical compounds suspected to be endocrine disruptor. In this study, 46 compounds exhibited antagonist activity at 50% of the maximum activity (IC x 50) within 11-9940 nM. In particular, 10,10-Oxybisphenoxarsine, triphenyltin fluoride, triphenyltin hydroxide, and chlorothalonil had strong hTR-antagonist activities. This knowledge gained from the present study will boost chemical regulation strategies for human and wildlife health. (C) 2022 The Author(s). Published by Elsevier Inc.

It is important to investigate contamination levels of neonicotinoide insecticides and the other pesticides that may cause adverse effects, not only to human health but also to the environmental ecosystems near rivers because of the large amounts of pesticides used for the paddy rice fields. Therefore, the runoff movements of commonly used 31 pesticides (12 fungicides, 4 neonicotinoid insecticides, 8 insecticides, and 7 related metabolites) in a small river basin, which branches from a major river, located near paddy rice fields in Japan were presented in this study. The concentration of the pesticide residues found in the present study fluctuated depending on the applied periods and amounts as well as the flow of the river water. No pesticides exceeded the standard levels in environmental and drinking water set by the government of Japan. The maximum loadings of total pesticides in the river were obtained in July for fungicides, August for neonicotinoids and May for the other insecticides. The monthly loading of total pesticides exhibited unique variations. The runoff ratios of neonicotinoids from paddy fields to the river ranged from 078% of imidacloprid to 10% for dinotefuran. Results obtained in this study are important for the evaluation of the actual behavior of the insecticides and fungicides in small rivers.

hER-MIP is a molecularly imprinted polymer (MIP) that has been shown to selectively collect human estrogen receptor (hER) binding active substances. However, environmental samples contain various chemicals depending on the location and regional differences, and the hER binding activity depends on the sample type. Thus, the general applicability of hER-MIP to actual environmental samples must be elucidated. In this study, 48 environmental samples were collected and screened with hER-MIP, and a yeast assay was performed to evaluate the adsorption characteristics of the samples according to the adsorption and elution fractions. The results showed that hER-MIP collects hER binding active substances almost selectively but does not collect constitutive androstane receptor (CAR) binding active substances selectively. CAR binding activity was detected in the adsorbed fraction because several hER binding active substances also demonstrate CAR binding activity.

A novel acidic morpholine-derivative containing glyceride (M-glyceride) was isolated from the cells of two strains of the thraustochytrid, Aurantiochytrium . The glyceride accounted for approximately 0.1 -0.4% of the lyophilized cells. The glyceride consisted of peaks I (85%) and II (15%). The structures of the intact and acetylated glycerides were elucidated by liquid chromatography-quadrupole time-of-flight chromatograph mass spectrometer (LC–Q/TOF) and NMR spectroscopy. The hydrate type of M-glyceride was detected as a minor component by LC–MS/MS. By 2D-NMR experiments, peaks I of the intact M-glyceride were elucidated as 1,2-didocosapentaenoyl-glyceryl-2′-oxy-3′-oxomorpholino propionic acid, and peak II was estimated 1,2-palmitoyldocosapentaenoyl- and/or 1,2-docosapentaenoylpalmitoyl-glyceryl-2′-oxy-3′-oxomorpholino propionic acid. The double bond position of docosapentaenoic acid was of the ω − 6 type (C 22 = 5.ω − 6). The M-glyceride content varied by the cell cycle. The content was 0.4% of lyophilized cells at the mid logarithmic phase, and decreased to 0.1% at the mid stationary phase. When cells were grown in 1.0 µM M-glyceride-containing growth media, cell growth was stimulated to 110% of the control. With 0.1 µM acetyl M-glyceride, stimulation of 113% of the control was observed. Finding morpholine derivatives in biological components is rare, and 2-hydroxy-3-oxomorpholino propionic acid (auranic acid) is a novel morpholine derivative.

There has been much effort to assess the efficacy of sewage treatment plants to remove chemicals with estrogenic activity, but other modes of toxicity have received less attention. This study assessed the efficacy of advanced tertiary technologies to remove estrogen receptor (ER), aryl hydrocarbon receptor (AhR), retinoic receptor (RAR) agonists and genotoxicity using recombinant gene bioassays. Untreated sewage contained ER, AhR and RAR agonists and was genotoxic. Activated sludge treatment removed over 69% of the ER, AhR and RAR aqueous phase activities. Ozonation and biologically activated carbon processes removed genotoxicity and most receptor agonists to below detection limits. Estrogenic activity was associated with the semi-polar fraction of the aqueous phase and non-polar phase of the influent (maximum 30%). AhR and RAR activities were found in both the aqueous and particulate phases of influent. Only RAR activity was frequently found in the polar fraction of aqueous and particulate phases of the process effluent. In the influent, the greatest RAR activity was found in the polar fraction of the aqueous phase (>65%). Genotoxic activity was detected in all influent fractions but rapidly decreased. The results showed that ER, AhR and RAR agonists were greatly reduced by the advanced tertiary treatment processes.

Odd-carbon fatty acids have been utilized for anaplerotic therapy for lifestyle-related diseases such as Alzheimer’s disease and diabetes. We examined the culture condition of Aurantiochytrium for the production of pentadecanoic acid (C15) as an odd-carbon fatty acid, and demonstrated a produce method for odd-carbon fatty acid–enriched triglyceride. To determine the optimum conditions for C15 production, selection of effective components and their respective concentrations in the culture medium, choice of the suitable Aurantiochytrium strain, and determination of the optimum culture pH were performed. The optimum conditions for the production of C15 were found to be as follows: Strain: Aurantiochytrium sp. strain SA-96 Medium composition: 0.2% yeast extract 0.5% monosodium glutamate, 1.0% sea salt, 50 mM L-Val, 25 mM sodium propionate, 3.6% glucose, and 10% soy milk whey. Culture system: Airlift culture vessels (5.0 L) and pH controller were used. Sodium hydroxide (1.0 M) solution was used for pH adjustment. Other parameters were as follows: medium volume, 3.0 L; air supply, 1.2 vvm; temperature, 23.5–26.6 °C; pH, 7.40–7.74; culture period, 72 h. The yield of odd-carbon fatty acids (C15, 85% of odd FA) was 1.04 g L−1. Preparation of odd-carbon fatty acid–enriched triglyceride: Triglyceride was isolated from the extracted lipids from cultured cells and treated with ozone/hydrogen peroxide. By removing of carboxylic acid containing triglyceride, the odd-carbon fatty acid and C15 contents of the resultant triglyceride reached to 72.3 and 60.7%, respectively.

Reproductive disorders in birds are the most characteristic effects of DDT contamination of wildlife. Experimental exposure of avian eggs to the estrogenic substance o,p'-DDT causes abnormal development of the reproductive tract (shortening of the left oviduct and aberrant development of the right oviduct) and eggshell thinning in mature birds, but it is still not known how eggshell thinning occurs in the abnormal oviduct. To fill this information gap, we examined the histology of the uterine part of the oviduct in Japanese quail treated in ovo with o,p'-DDT or a synthetic estrogen, diethylstilbestrol (DES), and we performed immunohistochemical staining for the calcium-binding proteins CALB1, SPP1, and TRPV6. Both o,p'-DDT-treated and DES-treated quail had few, and scattered, gland cells in the left uterus, unlike vehicle controls, in which gland cells tightly occupied the lamina propria. The aberrantly developed right uterus retained all the components of the normal left uterus, but in immature form. Immunostaining for CALB1, SPP1, and TRPV6 was greatly reduced by both o,p'-DDT and DES; SPP1 and TRPV6 immunostaining patterns, in particular, differed distinctly from those in the controls. These findings suggest that CALB1, SPP1, and TRPV6 are molecular factors, decreased production of which is responsible for eggshell thinning. Our findings also could contribute to understanding of the eggshell formation mechanism in birds.

Automated identification and determination system using a gas chromatograph-mass spectrometry (AIQS-GC) is a technology that enables semi-identification and quantification of substances without using the standard substance. Using this method, environmental water monitoring around the disaster temporary storage area after the Great East Japan Earthquake was conducted. Di (2-ethylhexyl) phthalate, pyrene, etc. were detected at high concentration. Some substances showed high concentration near the temporary storage area, suggesting the possibility of elution from disaster waste. Thus, AIQS-GC was effective in emergency monitoring such as disasters. The authors held several workshops to disseminate this system. We have also started joint research with the local environment research institute on the improvement of AIQS-GC. In this joint research, we have a plan to add the number of listed substances, expand the applicable equipment manufacturers, and study the inter-apparatus and inter-laboratory error. In addition, AIQS can be analyzed retrospectively, accumulated and aggregated measurement data is expected.

2011年3月11日の東日本大震災によって発生した災害廃棄物は, 地方公共団体が管理する土地を仮置き場として野積みにされた。2011年6月, 隣接する空き地に災害廃棄物が積み上げられた宮城県石巻市の高等学校を訪問し, 瓦礫から発生したと考えられるハエ類や浮遊微生物の調査を実施した。校舎内で捕獲したハエ類30個体のうち, 17個体は屋内侵入性のイエバエ類Muscoideaであった。また, ハエ類の体表面から病菌細菌の分離を試みたところ, 大腸菌Escherichia coliが30個体中15個体(50%)から, 緑膿菌Pseudomonas aeruginosaが5個体(17%)から, サルモネラSalmonella entericaが13個体(43%)から分離された。屋内の浮遊細菌および浮遊真菌濃度は日本建築学会の規準値以下であったが, 屋外の浮遊真菌濃度は災害廃棄物の瓦礫に近いほど高くなる傾向にあった。また, 吸入性アレルゲンとして重要な真菌であるAlternaria属およびEurotium属の屋外濃度が国内の一般環境と比較し, 高濃度であることが明らかになった。

東日本大震災は, 2011年3月11日に発生した東北地方太平洋沖地震による災害であり, 海岸線に押し寄せた津波によって甚大な被害をもたらした。津波被災地域の避難所等の居住区では, 津波堆積物等由来の飛散粉じんによる呼吸器疾患等の健康被害が懸念された。本調査では, 宮城県と連携して, 同年5月～9月にかけて, 津波被災地域の避難所5カ所と高等学校1カ所の室内浮遊粉じん(PM2.5, PM10及びTSP)の測定とハウスダストの性状評価及び元素分析を実施した。PM2.5の濃度は, 調査期間の平均値が18～31 μg/m3であり, 1日平均値の環境基準35 μg/m3以下であった。また, PM2.5, PM10及びTSPの濃度は, 調査期間を通じて概して低い値であり, 大半の測定場所で時間経過に伴って減少傾向であった。一方で, 人の活動が活発になる午前8～9時から午後4～5時においては, 35 μg/m3を超過することがあり, 高感受性者の体調の変化に注意を向ける必要があると考えられた。室内浮遊粉じんの測定結果は, 津波に浸水した建屋(或いはその近傍)かどうか, 土足で出入りしているかどうか, 定期清掃を行っているかどうかが, その濃度に影響を与える重要な因子であることを示唆した。ハウスダストのふるい処理による性状評価は, 避難所のハウスダストの観察によって室内浮遊粉じんの発生状況を推察できる可能性を示した。また, ハウスダストの元素分析の結果は, 避難所内に設置されている物品由来の化学物質に留意が必要であることを示した。

The possibility that a wide variety of polychlorinated biphenyls (PCBs) and their derivatives are present in the environment necessitates detailed evaluation of their toxicities. A number of PCBs and monohydroxylated PCBs (OH-PCBs) have been reported to have affinities for several nuclear receptors, and some of these compounds have been shown to have harmful effects in animals. In this study, we focused on the retinoid X receptor (RXR) and the retinoic acid receptor (RAR), which play critical roles in development and homeostasis. Specifically, we measured the agonistic activities of 91 OH-PCBs on these receptors by using yeast cells transfected with human RXRβ or RARγ. Of the tested OH-PCBs, 20 exhibited agonistic activity on RXRβ and 35 on RARγ, although their activities were lower than those of the respective endogenous ligands. Most of the OH-PCBs that showed potent activity on RXRβ were tri- or tetrachlorinated compounds, whereas most of the active compounds on RARγ were di-, tri-, or tetrachlorinated compounds. The optimal position for the hydroxyl group relative to the bond linking the aromatic rings was ortho for RXRβ and meta for RARγ. The activities of the OH-PCBs on RXRβ depended strongly on the position of the hydroxyl group, whereas those on RARγ did not. Taken together with the results of our previous studies of compounds with effects on other nuclear receptors, these results provide critical information for further research on receptor-mediated toxicity, endocrine-disrupting effects, and structure-activity relationships.

The Australian Antarctic Division (AAD) operates Australia's Davis Station in the Antarctic. In 2005, Davis Station's wastewater treatment plant failed and since then untreated, macerated effluent has been discharged to the ocean. The objectives of this study were to determine whether an advanced water treatment plant (AWTP) commissioned by the AAD and featuring a multi-barrier process involving ozonation, ceramic microfiltration, biologically activated carbon filtration, reverse osmosis, ultraviolet disinfection and chlorination was capable of producing potable water and a non-toxic brine concentrate that can be discharged with minimal environmental impact. The AWTP was tested using water from a municipal wastewater treatment plant in Tasmania, Australia. We used spot water and passive sampling combined with two multi-residue chromatographic-mass spectrometric methods and a range of recombinant receptor-reporter gene bioassays to screen trace organic chemicals (TrOCs), toxicity and receptor activity in the Feed water, in the environmental discharge (reject water), and product water from the AWTP for six months during 2014-15, and then again for three months in 2016. Across the two surveys we unambiguously detected 109 different TrOCs in the feed water, 39 chemicals in the reject water, and 34 chemicals in the product water. Sample toxicity and receptor activity in the feed water samples was almost totally removed in both testing periods, confirming that the vast majority of the receptor active TrOCs were removed by the treatment process. All the NDMA entering the AWTP in the feed and/or produced in the plant (typically < 50 ng/L), was retained into the reject water with no NDMA observed in the product water. In conclusion, the AWTP was working to design, and releases of TrOCs at the concentrations observed in this study would be unlikely cause adverse effects on populations of aquatic organisms in the receiving environment or users of the potable product water.

In pesticide risk management in Japan, predicted environmental concentrations are estimated by a tiered approach, and the Ministry of the Environment also performs field surveys to confirm the maximum concentrations of pesticides with risk concerns. To contribute to more efficient and effective field surveys, we developed the Pesticide Chemicals High Resolution Estimation Method (PeCHREM) for estimating spatially and temporally variable emissions of various paddy herbicides from paddy fields to the environment. We used PeCHREM and the G-CIEMS multimedia environmental fate model to predict day-to-day environmental concentration changes of 25 herbicides throughout Japan. To validate the PeCHREM/G-CIEMS model, we also conducted a field survey, in which river waters were sampled at least once every two weeks at seven sites in six prefectures from April to July 2009. In 20 of 139 sampling site-herbicide combinations in which herbicides were detected in at least three samples, all observed concentrations differed from the corresponding prediction by less than one order of magnitude. We also compared peak concentrations and the dates on which the concentrations reached peak values (peak dates) between predictions and observations. The peak concentration differences between predictions and observations were less than one order of magnitude in 66% of the 166 sampling site-herbicide combinations in which herbicide was detected in river water. The observed and predicted peak dates differed by less than two weeks in 79% of these 166 combinations. These results confirm that the PeCHREM/G-CIEMS model can improve the efficiency and effectiveness of surveys by predicting the peak concentrations and peak dates of various herbicides.

We screened 583 chemicals for receptor binding activity to the human estrogen receptor (hER), the Japanese medaka estrogen receptor (medER), and the aryl hydrocarbon receptor (AhR) using the yeast two-hybrid assay. The substances tested included substances that could potentially be produced unintentionally by industrial processes, such as halogenated steroids and phenols. Antagonistic effects on hER and the androgen receptor were also screened. The test chemicals were selected for screening on the basis of chemical structure associated with possible estrogen receptor binding activity. The current study presents the report on the screening of 583 chemicals for different kinds of endocrine disrupting activity.

We report an efficient screening procedure for the selective detection of compounds that are actively bound to estrogen receptor (ER) from environmental water samples using a receptor-mimic adsorbent prepared by a molecularly imprinted polymer (MIP). To mimic the recognition ability of ER, we improved the typical MIP preparation procedure using a hydrophilic matrix with a polyethylene glycol (PEG)-based crosslinker and a hydrophobic monomer to imitate the hydrophobic pocket of ER. An optimized MIP prepared with methacrylic acid as an additional functional monomer and estriol (E3), an analogue of 17 beta-estradiol (E2), exhibited highly selective adsorption for ER-active compounds such as E2 and E3, with significant suppression of non-specific hydrophobic adsorption. The prepared MIP was then applied to the screening of ER-active compounds in sewage samples. The fraction concentrated by the MIP was evaluated by in vitro bioassay using the yeast two-hybrid (Y2H) method and liquid chromatography -quadrupole time-of-flight mass spectrometry (LC-Q-TOFMS). Compared to an authentic adsorbent, styrene-divinylbenzene (SDB)-based resin, the fraction concentrated by the MIP had 120% ER activity in the Y2H assay, and only 25% peak volume was detected in LC-Q-TOFMS. Furthermore, a few ER-active compounds were identified only from the fraction concentrated by the MIP, although they could not be determined in the fraction concentrated by the SDB-based resin due to ion suppression along with high levels of hydrophobic compounds. These results indicated that the newly developed MIP effectively captured ER-active compounds and while allowing most non-ER-active compounds to pass through. (C) 2018 Elsevier Ltd. All rights reserved.

External exposure of workers engaging demolition works of damaged buildings performed near the Fukushima Daiichi Nuclear Power Plants was investigated. Indoor air dose rates recorded near the ceiling were higher than those recorded near the floor, and smaller than outdoor air dose rates. It was found that demolition works at “residence restricted area”, in some cases, might meet specified high dose rate works, because average air dose rate of a demolition work performed in this area exceeded 2.5 µSv/h. Average daily external exposure of workers at residence restricted area was 6.4 µSv, it was higher than that at “zone in preparation for the lifting of the evacuation order,” 4.4 µSv. It may reflect air dose rates recorded at these areas. On the other hands, average cumulative external exposure at “zone in preparation for the lifting of the evacuation order” was 146 µSv, it was higher than that at “residence restricted area,” 89 µSv. This tendency may reflect a long period of demolition work and less of the workers. In demolition works at “zone in preparation for the lifting of the evacuation order,” “removing roofing tiles” was selected as the work which significantly increased external exposures by multiple regression analysis. “Removing interior materials” was also selected as the work which significantly decreased external exposures in demolition works at “residence restricted area.”

The constitutive androstane receptor (CAR) is a nuclear receptor and transcription factor regulating proteins involved in xenobiotic metabolism. Agonist activation of the CAR can trigger metabolic activation and toxification as well as detoxification and clearance; accordingly, xenobiotic substances acting as CAR ligands may pose a threat to human and animal health. We used yeast cells transduced with the human CAR and the response pathway to measure the CAR-agonistic activities of 549 synthetic or natural compounds: 216 of the tested compounds exhibited CAR-agonistic effects. Eighty-four percent of CAR-activating compounds were aromatic compounds, and >65% of these active compounds were aromatic hydrocarbons, bisphenols, monoalkyl phenols, phthalates, styrene dimers, diphenyl ethers, organochlorines, and organophosphates. The ten most potent compounds were 4-tert-octylphenol (4tOP; reference substance), 4-nonylphenol, diethylstilbestrol, benzyl n-butyl phthalate, 2-(4-hydroxyphenyl)-2,4,4-trimethylchroman, o,p'-DDT, methoxychlor, di-n-propyl phthalate, hexestrol, and octachlorostyrene. The activities of these nine non-reference compounds exceeded 10% of the 4tOP activity. Analysis of para-monoalkyl phenols suggests that branching of the alkyl group and chlorination at the ortho position raises potency. This study provides critical information for identifying the potential of CAR-mediated toxic hazards and for understanding the relevant mechanism.

Instrumentation and monitoring systems in a nuclear power plant are very important to monitor plant conditions for safe operations and a plant shutdown. The severe accident at TOKYO ELECTRIC POWER COMPANY's Fukushima Daiichi Nuclear Power Station (hereinafter called as TF1) in March 2011 caused several severe situations such as core damage, hydrogen explosion, etc. Lessons learned from the severe accident at TF1 show that an appropriate operable instrumentation and monitoring system for a severe accident should be developed so that the system will deliver an appropriate performance for mitigation of severe accident condition in a nuclear power plant. This paper proposes the classification method of severe accident condition for the development of an appropriate operable instrumentation and monitoring system for a severe accident based on the problem analysis of monitoring variables during the severe accident at TF1. The classification is formed on the basis of the integrity of boundary for plant safety and the successful (or unsuccessful) condition of the cooling water injection, and is used for an establishment of defining severe accident environmental conditions for the instrumentation and monitoring system. Examples of the establishment method are also shown in this paper.