Molecular characterization and developmental expression patterns of thyroid hormone receptors (TRs) and their responsiveness to TR agonist and antagonist in Rana nigromaculata

Considering some advantages of Rana nigromaculata as an experimental species, we propose that this species, like Xenopus laevis, could be used to assay thyroid hormone（TH） signaling disrupting actions. To validate the utilizability of R. nigromaculata, we investigated the responsiveness of R. nigromaculata to a TH receptor（TR） agonist（T3） and antagonist（amiodarone） by analyzing expression, based on characterizing TR cDNA and developmental expression patterns. With high levels of identity with the corresponding genes in X. laevis, both TRα and TRβ in R. nigromaculata exhibited roughly similar developmental expression patterns to those of X. laevis, in spite of some species-specific differences. Both TRα and TRβ expression had greater changes in the liver and intestine than in the tail and brain during metamorphosis. T3 exposure for 2 days induced more dramatic increases of TRβ expression in stage 27 than in stage34 tadpoles but not in stage 42 tadpoles, showing that the responsiveness of R. nigromaculata to TH decreased with development and disappeared at the onset of metamorphic climax.Corresponding to greater changes of TRβ expression in the liver and intestine than in the tail and brain during metamorphosis, the liver and intestine had higher responsiveness to exogenous T3 than the tail and brain. Amiodarone inhibited T3-induced TRβ expression. Our results show that R. nigromaculata can be used as a model species for assaying TH signaling disrupting actions by analyzing TRβ expression, and intestine tissues at stage 27 are ideal test materials due to high responsiveness and easy accessibility.     

Effects of biosurfactants on assays of PCB congeners in transgenic arabidopsis plants carrying a recombinant guinea pig AhR-mediated GUS reporter gene expression system

The transgenic Arabidopsis plants carrying a recombinant guinea pig (g) aryl hydrocarbon receptor (AhR)-mediated β-glucuronidase (GUS) reporter gene expression system were generated for assays of polychlorinated biphenyl (PCB) congeners. The selected transgenic Arabidopsis plant XgD2V11-6 exhibited a correlation between uptake of PCB126 and PCB126-induced GUS activity. Also, the plants showed induced GUS activity towards the supplemental indole 3-acetic acid (IAA). Thus, the GUS assay may reflect induction by both endogenous and exogenous AhR ligands. When biosurfactants, MEL-B, produced in the culture of yeast isolated from plants were used for assays of PCB congeners in the transgenic Arabidopsis plants, they showed marked PCB126 dose-dependent and toxic equivalency factor (TEF) dependent GUS activities. The effects of biosurfactants were clearer when the plants were cultivated on soils containing PCB congeners for 7 days as compared with on soils for 3 days as well as in the medium for 3 days. Threfore, it was estimated that biosurfactants form micellae with PCB congeners, which are easily uptaken by the plants in a mode of passive diffusion, transport into the aerial parts and then induce GUS activity.     

Dioxin-like activity in environmental and human samples from Greenland and Denmark

Dioxins and dioxin-like (DL) compounds are some of the most toxic chemicals being highly persistent in the environment. The toxicological effects of dioxins are mediated via the aryl hydrocarbon receptor (AhR). Compounds of diverse structure and lipophility can bind and activate AhR. The AhR transactivation bioassay is utilized in an array of projects to study the AhR-mediated activities of individual chemicals and mixtures and for epidemiological purposes.This review summarizes a series of studies regarding the DL-activity of single compounds and complex compound mixtures in the environment and humans.We found that some pesticides, plasticizers and phytoestrogens can activate the AhR, and the combined effect of compounds with no or weak AhR potency cannot be ignored.The significant DL-activity in the wastewater effluent indicates the treatment is not sufficient to prevent contamination of surface waters with dioxins. Our results from human studies suggest that the serum DL-activity reflect the complex mixture of persistent organic pollutants (POPs). Greenlandic Inuit had lower serum DL-activity level compared to Europeans, probably due to long distance from the dioxin sources and UV degradation of the high potent dioxin and/or the inhibitory effect of the high level of non-DL POPs. Selective bioaccumulation of PCBs in the food chain may contribute to the negative correlation between serum POPs and DL-activity observed in Greenlandic Inuit.Hence the AhR transactivation bioassay provides a cost-effective and integrated screening tool for measurement of the DL-activity in human, environmental and commercial samples.     

Integral assessment of estrogenic potentials in sediment-associated samples

Background, aim, and scope  The enzyme-linked receptor assay (ELRA) detects estrogenic and anti-estrogenic effects at the molecular level of receptor binding and is a useful tool for the integrative assessment of ecotoxicological potentials caused by hormonally active agents (HAA) and endocrine disrupting compounds (EDC). The main advantage of the ELRA is its high sample throughput and its robustness against cytotoxicity and microbial contamination. After a methodological adaptation to salinity of the ELRA, according to the first part of this study, which increased its salinity tolerance and sensitivity for 17-β-estradiol, the optimised ELRA was used to investigate 13 native sediments characterised by different levels of salinity and chemical contamination. The applicability of the ELRA for routine analysis in environmental assessment was evaluated. Salinity is often a critical factor for bioassays in ecotoxicological sediment assessment. Therefore, salinity of the samples was additionally adjusted to different levels to characterise its influence on elution and binding processes of receptor-binding substances. Materials and methods  The ELRA was carried out with the human estrogen receptor α (ER) in a 96-well microplate format using the experimental setup known from the competitive immunoassay based on ligand–protein interaction. It is an important improvement that a physiologically relevant receptor was used as a linking protein instead of an antibody. The microplates were coated with a 17-β-estradiol-BSA conjugate, and dilution series of estradiol and of native sediment samples were added and incubated with the ER. After a washing step, a biotinylated mouse anti-ER antibody was added to each well. Receptor binding to estradiol, agonistic and antagonistic receptor binding, were determined by a streptavidin-POD-biotin complex with subsequent measurement of the peroxidase activity at the wavelength of 450 nm using a commercial ELISA multiplate reader. The sediment elutriates and pore water samples of sediments were tested in a dilution series to evaluate at which dilution step the receptor-binding potential ends. In the elution process (see Section 2.1 to 2.2), a method was developed to adjust the salinity to the levels of the reference testings, which offers an appropriate option to adjust the salinity in both directions. Statistical evaluation was made with a combination of the Mann–Whitney U test and the pT-method. Results  This part of the study characterised the environmental factor ‘salinity’ for prospective applications of the ELRA. Using reference substances such as 17-β-estradiol, the ELRA showed sigmoid concentration-effect relations over a broad range from 0.05 μg/l to 100 μg/l under physiological conditions. After methodological optimisation, both sensitivity and tolerance of the assay against salinity could be significantly raised, and the ELRA became applicable under salinity conditions up to concentrations of 20.5‰. The mean relative inter-test error (n = 3) was around 11% with reference substances and below 5% for single sediments elutriates in three replicates each. For sediment testings, the pore water and different salinity-adjusted elutriates of 13 sediments were used. A clear differentiation of the receptor-binding potential could be reached by application of the pT-method. Thereby, pT-values from one to six could be assigned to the sediments, and the deviation caused by the different salinity conditions was one pT-value. The mean standard deviation in the salinity adaptation procedure of the elutriates was below 5%. Discussion  Although the ELRA has already been used for assessments of wastewater, sludge and soil, its applicability for samples to different salinity levels has not been investigated so far. Even if the ELRA is not as sensitive as the E-screen or the YES-assay, with regard to reference substances like 17-β-estradiol, it is a very useful tool for pre-screening, because it is able to integrate both estrogenic as well as anti-estrogenic receptor-binding effects. According to the results of sediment testing, and given the integrative power to detect different directions of effects, the ELRA shows sufficient sensitivity and salinity tolerance to discriminate receptor-binding potentials in environmental samples. Conclusions  The optimised ELRA assay is a fast, cost-effective, reliable and highly reproducible tool that can be used for high-throughput screening in a microplate format in detecting both estrogenic and anti-estrogenic effects. Additionally, the ELRA is robust against microbial contaminations, and is not susceptible towards cytotoxic interferences like the common cell-culture methods. The general applicability and sufficient sensitivity of the ELRA was shown in freshwater environments. Marine and brackish samples can be measured up to salinity levels of 20.5‰. Recommendations and perspectives  In view of the proven sensitivity, functionality and the fastness of the ELRA, it is recommendable to standardise the test method. At the moment, no adequate in vitro test procedure exists which is standardised to DIN or ISO levels. The E-screen and the yeast estrogen/androgen screens (YES/YAS) sometimes underlie strong cytotoxic effects, as reported in the first part of this study. Further development of an ELRA assay using human androgen receptors appears to be very promising to gain information about androgenic and anti-androgenic effects, too. This would offer a possibility to use the ELRA as a fast and reliable pre-screening tool for the detection of endocrine potentials, thus minimising time and cost-expensive animal experiments.     

好氧反硝化菌的脱氮特性研究与探讨

本论文对好氧反硝化菌株FR2,FX5,FW4在好氧条件下的反硝化特性进行了研究。结果表明：反硝化主要发生在菌体的对数生长期,且反硝化过程中NO2^--N有少量的积累。 对不同温度、pH、碳源对好氧反硝化作用的影响进行了研究,结果表明：该三株菌在好氧条件下均具有良好的反硝化效果;温度为25℃～30℃的中性和微碱性条件反硝化效果较好;以乙酸钠为碳源的反硝化效果最好,NO3^-N的去除率最高。以葡萄糖为碳源的反硝化效果最差,NO3^-N的去除率最低。     

基于绝对主成分-多元线性回归的滇池污染源解析

定量解析污染源是湖泊流域水环境管理的重要基础.基于滇池草海和外海多年水质监测数据,采用主成分分析（PCA）方法识别了主要水质指标的污染源类型,利用绝对主成分-多元线性回归模型（APCS-MLR）得到不同污染源对水质的贡献程度.结果表明,草海主要的污染源有农业面源、城市面源和内源3类,外海的主要污染源是农业面源、城镇生活污染源、城市面源和内源4类.与河流水污染源解析结果不同,底泥内源与气象因子对滇池主要水质指标的影响较大.     

Influence of rice straw burning on the levels of polycyclic aromatic hydrocarbons in agricultural county, Taiwan

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经饮水亚慢性铝染毒对大鼠血铁及相关蛋白水平的影响

为了探讨慢性铝暴露对大鼠铁代谢的影响,将100只4周龄清洁级雄性Wistar大鼠随机均分成摄铝组(430mg·L-1,以Al3+计)与对照组(蒸馏水),饮水染铝,每隔30d处死染铝大鼠和对照大鼠各10只,实验周期为150d.同时,在不同时间点(30d、60d、90d、120d、150d)测量大鼠体重,并进行协方差分析.最后,用火焰原子吸收分光光度法、固相竞争ELISA和比色法检测血浆Al、Fe、转铁蛋白(TF)、可溶性转铁蛋白受体(sTfR)含量及总铁结合力(TIBC).结果表明,在摄铝前期(90d前),染铝对大鼠体重无显著影响,而后期(120d后)染铝对大鼠体重抑制作用较大.在实验周期内,各时间点间对照组血浆中各指标均无显著差异,摄铝组随着时间的延长各指标变化较大.各时间点摄铝组血浆中Al含量及Al/Fe均显著高于对照组(p0.01);30d时摄铝组血浆中Fe含量与对照组相比无显著差异(p0.05),60、90、120d时均显著低于对照组(p0.01),150d时显著高于对照组(p0.01);30、60d时摄铝组血浆TF含量与对照组相比无显著差异(p0.05),90d后显著高于对照组(p0.05;p0.01);30、60d时摄铝组TIBC与对照组相比无显著差异(p0.05),其后各时间点均显著高于对照组(p0.01);30d时摄铝组sTfR含量与对照组相比无显著差异(p0.05),其后各时间点均显著低于对照组(p0.01).由此可见,铝可干扰大鼠体内铁的代谢,影响铁的生物学作用.     

基于气团老化程度对挥发性有机物分类改善PMF源解析效果

VOCs作为臭氧与二次有机气溶胶的关键前体物,其来源解析对臭氧和颗粒物的协同控制至关重要.但多数VOCs具有反应性,不能完全满足受体模式对污染源排放化学物质组成稳定的假设要求,导致受体模式解析结果不能精准反映实际源贡献.为解决因不同VOCs反应活性不同而导致的组分相对变化与模型假设不符的问题,引入VOCs老化程度来表征其光化学反应的状态.将乌海市超级观测站监测的VOCs依据乙苯与间/对-二甲苯的比值分为高、中和低这3种老化状态.结果表明,分类后解析结果的模拟值与实测值的回归方程参数(斜率和截距)、标准误差、决定系数和残差合格率等模型诊断参数改善明显.因为老化程度与气团在大气中的传输时间和大气氧化性密切相关,在一定程度上也反映了气团的不同来源信息.在高老化态样品中焦化源贡献率最高,达47.20%;在低老化态样品中燃烧源和焦化源均较高,分别为28.67%和24.39%.按老化程度进行分类后,PMF对VOCs源解析的结果更符合实际排放源的贡献.     

Prenatal diagnosis of Apert syndrome: report of two cases

Two de novo cases with Apert Syndrome detected prenatally are presented herein. In the first, fetal ultrasound findings of syndactyly of the hands, craniosynostosis and proptosis resulted in a prenatal diagnosis in the nineteenth week of gestation. This is the earliest prenatal diagnosis of this syndrome in a not-at-risk case. Following counseling, this pregnancy was terminated and subsequent pathological examination and DNA analysis confirmed the diagnosis of Apert Syndrome and coarctation of the aorta. In the second case, fetal ultrasound at 21 weeks' gestation revealed a hypoplastic left heart and clover-leaf skull. Following counseling, this pregnancy was also terminated. Further examination of the fetus and DNA analysis led to a diagnosis of Apert Syndrome. These cases emphasize the need to complete a thorough fetal ultrasound in cases with potentially lethal cardiac abnormality and the importance of incorporating a fetal pathologist, as well as a medical geneticist, in the investigations performed after delivery or pregnancy termination when a fetal abnormality is detected on ultrasound. Copyright © 2003 John Wiley & Sons, Ltd.