Assays of PCB congeners and organochlorine insecticides with the transgenic Arabidopsis and tobacco plants carrying recombinant guinea pig AhR and GUS reporter genes

Certain congeners of polychlorinated biphenyls (PCBs) and organochlorine insecticides are ligands of aryl hydrocarbon receptors (AhRs) in animals. A recombinant guinea pig (g) AhR, XgDV, was constructed by fusing the ligand-binding domain of gAhR, the DNA-binding domain of LexA, and the transactivating domain of VP16. Then, the expression unit of β-glucuronidase (GUS) reporter gene regulated by XgDV was introduced into Arabidopsis and tobacco plants. When the transgenic Arabidopsis XgDV plants were cultured on Murashige–Skoog (MS) medium containing PCB congeners, the GUS activity in the plants increased toxic equivalent (TEQ)-dependently. The GUS activity in the transgenic Arabidopsis XgDV plants cultured on MS medium containing the organochlorine insecticide dieldrin was also induced. On the other hand, in the case of DDT, the GUS activity induced by 3-methylcholanthere in the plants decreased. The transgenic Arabidopsis XgDV plants detected 1000 ng g^?1 PCB126 in 1 g of soils. Thus the XgDV plants seemed to be useful for convenient assays of PCB congeners and organochlorine insecticides, without any extraction and purification steps.     

The constitutively active Ah receptor (CA-AhR) mouse as a model for dioxin exposure - effects in reproductive organs

The dioxin/aryl hydrocarbon receptor (AhR) mediates most toxic effects of dioxins. In utero/lactational exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) impairs fetal/neonatal development and the developing male reproductive tract are among the most sensitive tissues. TCDD causes antiestrogenic responses in rodent mammary gland and uterus and in human breast cancer cell lines in the presence of estrogen. Also, more recently an estrogen-like effect of TCDD/AhR has been suggested in the absence of estrogen. A transgenic mouse expressing a constitutively active AhR (CA-AhR) was developed as a model mimicking a situation of constant exposure to AhR agonists. Male and female reproductive tissues of CA-AhR mice were characterized for some of the effects commonly seen after dioxin exposure. Sexually mature CA-AhR female mice showed decreased uterus weight, while an uterotrophic assay in immature CA-AhR mice resulted in increased uterus weight. In immature mice, both TCDD-exposure and CA-AhR increased the expression of the estrogen receptor target gene Cathepsin D. When co-treated with 17β-estradiol no increase in Cathepsin D levels occurred in either TCDD-exposed or CA-AhR mice. In sexually mature male CA-AhR mice the weights of testis and ventral prostate were decreased and the epididymal sperm reserve was reduced. The results of the present study are in accordance with previous studies on dioxin-exposed rodents in that an activated AhR (here CA-AhR) leads to antiestrogenic effects in the presence of estrogen, but to estrogenic effects in the absence of estrogen. These results suggest the CA-AhR mouse model as a useful tool for studies of continuous low activity of the AhR from early development, resembling the human exposure situation.     

Determination of dioxin concentrations in fish and seafood samples using a highly sensitive reporter cell line, DR-EcoScreen cells

There is a strong need for the development of relatively rapid and low-cost bioassays for the determination of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), and dioxin-like polychlorinated biphenyls (dl-PCBs) in environmental and food samples. In this study, we applied a reporter gene assay using DR-EcoScreen cells (DR-cell assay), which is highly sensitive to dioxins, to the determination of PCDD/Fs and dl-PCBs in fish and seafood samples. The PCDD/Fs and dl-PCBs were extracted from homogenated samples (10 g) of 30 fish and shellfish, purified by clean-up procedure using a multilayered silica gel column and an alumina column, and applied to DR-cell assay. Interestingly, the bioanalytical equivalent (BEQ) values obtained from the DR-cell assay [<0.1 ∼ 5.4 pg BEQ g⁻¹ wet weight (ww)] were closely correlated with the toxicity equivalent (TEQ) values from conventional high-resolution gas chromatography/high-resolution mass spectrometry (HRGC-HRMS) analysis (r² = 0.912), and the slope of regression line was 0.913. Therefore, we multiplied the BEQ values from the DR-cell assay by a conversion coefficient (1.095, the reciprocal of 0.913) to approximate the TEQ values from the HRGC-HRMS analysis. Furthermore, we used this DR-cell assay to perform a prescreening test of PCDD/Fs and dl-PCBs in 16 fish and seafood samples purchased from a supermarket, revealing that a sample from the fatty flesh of a bluefin tuna exceeded 8 pg TEQ g⁻¹ ww (the European Union-tolerance limit). Taken together, these results suggest that the DR-cell assay might be applicable as a rapid and low-cost prescreening method to determine dioxin levels in fish and seafood samples.     

Estimating receptor sensitivity to spatial proximity of emissions sources

Spatial proximity of emissions sources to receptors may affect sensitivity to potential adverse human health effects. This research investigates whether receptor sensitivity to the location of emission sources can be utilized efficiently to minimize health risk in selecting sites for industrial enterprises, thermal electric stations, etc. A sensitivity function that is independent of the location of pre-existing emission sources is derived and applied to Minsk, Belarus. The function estimates exposures based on weather and climatic conditions as well as the distribution of population density at a given locality. Arraying prospective sites based on their sensitivity function magnitude provides a technique for minimizing health risk based on receptor sensitivity to the spatial proximity of atmospheric emissions sources.     

酵母双杂交技术构建重组人雌激素受体基因酵母

应用酵母双杂交技术构建重组人雌激素受体基因酵母,用以检测类/抗雌激素化合物和环境样品的类/抗雌激素活性.采用多聚酶链反应(PCR)法扩增人雌激素受体(hER)基因,构建诱饵质粒pGBKT7-ER LBD;分别提取并纯化两种含有ER共激活因子基因的靶质粒pGAD424-GRIP1和pGAD424-SRC1;诱饵质粒和靶质粒同时转染酵母细胞Y187,于营养缺陷型培养基(SD/-Trp/-Leu)上筛选阳性菌落,分别构建两种ER双杂交酵母ER+GRIP1和ER+SRC1.考察双杂交酵母与不同激素:17β-雌二醇(E2)、二氢睾酮(DHT)、孕酮(PG)以及三碘甲状腺原氨酸(T3)的结合情况,并考察了雌激素受体拮抗剂4-羟基他莫昔芬(4-OHT)与酵母的相互作用.结果表明:ER+GRIP1和ER+SRC1酵母均能够专一性的和E2结合,并存在显著的剂量-效应关系,E2对ER+GRIP1和ER+SRC1酵母β-半乳糖苷酶活性诱导的EC50值分别为7.3×10-11mol·L-1和1.5×10-10mol·L-1,其中ER+GRIP1酵母细胞诱导产生的酶活性值明显高于ER+SRC1酵母细胞.4-OHT能够抑制E2诱导ER+GRIP1酵母细胞产生的酶活性,并存在显著的剂量-效应关系,IC50值为1.0×10-7mol·L-1.表明重组人雌激素受体基因酵母可以用于检测化合物和环境样品的类/抗雌激素活性.     

Reliability of Indicators of Decline in Abundance

Abstract: Although there are many indicators of endangerment (i.e., whether populations or species meet criteria that justify conservation action), their reliability has rarely been tested. Such indicators may fail to identify that a population or species meets criteria for conservation action (false negative) or may incorrectly show that such criteria have been met (false positive). To quantify the rate of both types of error for 20 commonly used indicators of declining abundance (threat indicators), we used receiver operating characteristic curves derived from historical (1938–2007) data for 18 sockeye salmon (Oncorhynchus nerka) populations in the Fraser River, British Columbia, Canada. We retrospectively determined each population's yearly status (reflected by change in abundance over time) on the basis of each indicator. We then compared that population's status in a given year with the status in subsequent years (determined by the magnitude of decline in abundance across those years). For each sockeye population, we calculated how often each indicator of past status matched subsequent status. No single threat indicator provided error‐free estimates of status, but indicators that reflected the extent (i.e., magnitude) of past decline in abundance (through comparison of current abundance with some historical baseline abundance) tended to better reflect status in subsequent years than the rate of decline over the previous 3 generations (a widely used indicator). We recommend that when possible, the reliability of various threat indicators be evaluated with empirical analyses before such indicators are used to determine the need for conservation action. These indicators should include estimates from the entire data set to take into account a historical baseline.     

安徽典型硫铁矿集中开采区土壤重金属污染特征及来源解析

为查明安徽某典型硫铁矿集中开采区周边土壤重金属元素污染特征及污染来源,采集了研究区42个表层土壤样品和2个垂向剖面的16个土壤样品测定其pH值及Zn、Cu、Cd、Pb、Cr、Ni、Hg和As重金属元素含量,运用ArcGIS反距离权重插值法、内梅罗综合指数法、主成分分析法和绝对因子得分-多元线性回归（APCS-MLR）受体模型等方法进行了重金属元素分布特征分析、污染评价和来源解析.结果表明,土壤中8种重金属元素的含量平均值是铜陵地区背景值的1.03~13.14倍,Zn、Cu、Cd、Pb、Hg和As这6种重金属元素局部富集明显,在空间分布上与采矿活动位置基本一致.单因子污染指数评价显示,Zn、Cu、Cd、Pb和As这5种元素存在不同程度的污染风险,Cd和Cu元素污染程度最为严重,中度污染以上占比分别为47.62%和42.86%;内梅罗综合污染指数评价显示,研究区土壤样点中度污染以上占比为61.90%;地累积指数评价显示,研究区Zn、Cu、Cd、Pb、Hg和As这6种重金属元素污染程度主要为轻污染~强污染.在土壤剖面垂向变化上,重金属元素易在土壤表层富集,向深部迁移较缓慢,高含量主要在1 m以上位置.源解析表明,Zn、Pb和As的来源中地质高背景和采矿活动贡献率分别为37.82%、43.49%和46.63%,成土母质风化的自然来源贡献率分别为34.02%、40.88%和38.52%;Cr和Ni来源主要为成土母质风化的自然来源,贡献率分别为91.95%和73.68%;Cu的来源中地质高背景和采矿活动贡献率为41.91%,大气沉降和农业综合源贡献率为41.30%;Hg的来源较多,成土母质风化的自然来源贡献率为35.60%,地质高背景和采矿活动贡献率为29.87%,未知来源贡献率为34.05%;Cd的主要来源为大气沉降和农业综合源,贡献率为81.81%.     

城市间PM2.5源的碳成分谱比较

为了解我国不同城市PM_2.5源的碳成分谱特征和地域差异,采集沈阳市、十堰市和乌鲁木齐市的燃煤源、柴油车尾气源、汽油车尾气源和餐饮源样品,使用热光透射法分析PM_2.5中的总碳（TC）、有机碳（OC）和元素碳（EC）,以及细分的8种碳组分（OC1,OC2,OC3,OC4,EC1,EC2,EC3和OPC_T）,构建各类污染源碳成分谱.结果表明：3个城市4类源TC/PM_2.5从高到低分别为：餐饮源（65.1%±8.4%）、柴油车尾气源（46.2%±9.5%）、汽油车尾气源（37.7%±3.5%）和燃煤源（17.3%±8.0%）;OC/TC在餐饮源中最高（98.0%±0.5%）,EC/TC在柴油车尾气源中最高（38.6%±8.5%）.3个城市同类源的碳组分含量受污染源细分后的不同类型影响有一定差异,但归一化处理后总体仍表现为燃煤源中OC2（14%~30%）和OC3（13%~23%）含量最高,柴油车尾气源中EC2（22%~56%）含量最高,汽油车尾气源中OC2（24%~41%）、OC1（16%~42%）和OC3（12%~26%）含量最高,餐饮源中OC2（21%~43%）和OC3（23%~49%）含量最高.不同污染源的OC/EC值为燃煤源在0.4~7.6之间,柴油车尾气源在0.2~5.6之间,汽油车尾气源在1.1~38.5之间,餐饮源在6.4~170.2之间.分歧系数结果显示3个城市不同源的碳成分谱具有差异性,同类源的碳成分谱具有相似性.将3个城市同类源碳成分谱合并后利用化学质量平衡灵敏度矩阵得到OC2,OC3,OC4,EC1和OPC_T可共同作为燃煤源的标识组分;EC2是柴油车尾气源的标识组分;OC1,OC2和OC3可共同作为汽油车尾气源的标识组分;OC2和OC3可共同作为餐饮源的标识组分.沈阳市、十堰市和乌鲁木齐市相同污染源相似的碳成分谱和一致的标识碳组分可为国内其他城市相关研究提供数据参考.     

新疆干旱区某矿冶场对周围土壤重金属累积的影响

我国西部干旱半干旱地区为生态脆弱区,近年来西部工矿业发展引起的土壤污染问题越来越受到关注.本文通过空间布点采样,采用多元统计分析、GIS技术以及地积累指数评估等手段,对新疆某矿冶场区周围土壤的9种重金属污染特征进行了调查.结果表明,Cu和As元素为研究区域的主要重金属污染物;重金属Cr、Zn、Ni和Cd累积程度较轻、受人为干扰小;Co和Pb元素在少数样点存在人为干扰累积的现象,但程度也较轻;土壤中Mn没有明显累积和人为干扰.因子分析结果发现9种重金属主要存在2种来源,As、Cu、Ni、Cr、Zn、Cd和Co主要来源于第1种源;元素Mn和Pb主要来源于第2种源.从重金属空间分布特征可知,As、Cu、Ni、Cr、Zn、Pb、Cd和Co的最高含量样点都分布在尾砂库附近区域,因此对尾砂库进行封闭处理是减少重金属污染物向周围环境扩散的重要措施.此外,本研究还发现基于大气污染分析的PCA/APCS受体模型不适用于本案例中定量分析土壤重金属来源.