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.     

Microscale Spatial Variation in Forest Litter Phytotoxicity

The spatial variation (within a 100 × 100 m plot) in the pollution of forest litter with heavy metals (Cu, Cd, Pb, and Zn), its acidity, and phytotoxicity (measured by the results of the root test using seedlings from a genetically homogeneous sample of common dandelion (Taraxacum officinale s.l.) have been estimated. Forest litter has been sampled in three zones differing in the toxic impact of long-term polymetal pollution by emissions from a copper-smelting plant emissions in the Middle Urals. The phytotoxicity variation is maximum in a moderately polluted plot, where both very high and very low pollution levels were observed, which determines a substantially nonlinear dose–effect relationship. The litter phytotoxicity is mainly accounted for by exchangeable forms of metals. Biological testing of samples from the most polluted plot has demonstrated marked antagonism between heavy metals and acidity.     

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.     

代谢活化作用对多溴代联苯类污染物类/抗甲状腺激素活性的影响

选用大鼠肝匀浆(S9)和鼠肝癌细胞(H4ⅡE)两种体系代谢活化多溴代联苯醚混标(BDEs)和十溴联苯醚(BDE209),采用重组甲状腺激素受体基因酵母检测BDEs和BDE209母体及其代谢产物的类/抗甲状腺激素效应.结果表明,BDEs和BDE209母体均不表现甲状腺激素效应(p>0.05);但是经S9和H4ⅡE细胞代谢活化后,其代谢产物表现出明显的类甲状腺激素活性和抗甲状腺激素活性(p<0.05),BDEs和BDE209的干扰甲状腺激素效应需要经过代谢活化步骤.比较不同代谢活化体系,重组酵母细胞本身的代谢活化作用并不显著,而H4ⅡE细胞和S9代谢活化体系均能够导致活性中间体.     

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.     

纳滤膜对脂肪族及杂环有机物的截留特性

分别选用NF90、NF270和NF-型号的纳滤膜测定了20种脂肪族及杂环有机物的截留率®,并分析了纳滤膜截留效果的影响因素.结果表明,脂肪族及杂环有机物的R受到分子结构和膜特性的影响:对于同分异构体,分枝结构越多,R越高;环状有机物与分子量相近的直链有机物相比,R明显偏高;孔径越小的纳滤膜R越高.利用遗传算法(GA)结合偏最小二乘回归(PLS)和人工神经网络法(ANN)建立了脂肪族及杂环物质的R与其结构的定量关系模型,2种方法建立的模型相关系数可分别达到0.8809和0.9944,通过2种模型进一步分析了R的影响规律,并对几种物质的R进行了有效的预测.从预测结果来看,GA-ANN模型的预测精度要好于GA-PLS模型.     

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.     

青藏高原表土重金属污染评价与来源解析

为了解青藏高原表层土壤重金属的污染特征、空间分布及污染来源,沿东北-西南方向对青藏高原表土(0～20 cm)样品进行了采集,对土壤中的Ba、Cd、Co、Cr、Cu、Fe、Mn、Mo、Ni、Pb、Sb、Sc和Zn等13种重金属总量进行了分析,并利用主成分分析-绝对主成分分数-多元线性回归(PCA-APCS-MLR)受体模型初步定量解析了重金属的潜在来源.结果表明Cd和Sb均显著超标,分别是20世纪70年代青藏高原土壤背景值的2. 13与1. 52倍.富集因子(EF)、地累积指数(I_(geo))和Nemero综合指数(PN)分析同样表明青藏高原表土主要以Cd和Sb污染为主,但污染程度普遍不高.在空间分布方面,青藏高原中部、东南部及东北部均呈不同程度污染,但中部及东南部污染相对较重. PCA-APCS-MLR分析表明,青藏高原土壤重金属主要有3个来源,分别为自然、交通和采矿等综合因素. Co、Cr、Cu、Fe、Mn、Ni和Sc主要受自然因素影响,Ba、Cd、Mo和Pb主要受交通因素影响,Zn主要受自然和交通因素共同影响,Sb主要受采矿、自然和交通等综合因素影响.青藏高原土壤重金属污染防治过程中应着重考虑受交通、采矿等综合因素影响下的Cd和Sb污染.     

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%.