Determination of estrogenic activity by LYES-assay (yeast estrogen screen-assay assisted by enzymatic digestion with lyticase)

In order to enhance the sensitivity and the speed of the yeast estrogen screen (YES)-assay, which has been established in many laboratories for the determination of estrogenic activity of compounds and environmental samples, the LYES-assay, a modified version of the YES-assay including a digestion step with the enzyme lyticase, was developed. With the LYES-assay the estrogenic activities of natural (17β-estradiol E2 and estrone), synthetic (17-ethinylestradiol EE2) and pharmaceutical estrogens (diethylstilbestrol DES) as well as xenoestrogens (4-nonylphenol NP and five parabens) were determined and compared with the results obtained by other in vitro-assays namely the conventional YES-assay, the E-Screen-assay (MCF-7 breast tumor cell proliferation) and a receptor binding-assay (RB) with human estrogen receptors hER- and hER-β. In the case of E2 the LYES-assay had a significantly lower limit of quantification (LOQ) than the conventional YES-assay and even two orders of magnitude lower than the RB-assay. Compared to the E-Screen-assay the LOQ of the LYES-assay was almost one order of magnitude higher. The time required to perform the LYES-assay was as little as seven hours compared to three to five days for the conventional YES-assay. Thus, the LYES-assay is a very good alternative to existing estrogenic in vitro-assays, since it has a good sensitivity, is cheap and much faster than the other assays.     

Toxic effects of selenium on marine fish

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Enhanced photocatalytic activity of nanotube-like titania by sulfuric acid treatment

IntroductionTiO2 nanotubehaswidelydrawnmuchattentionduetoitslargesurfaceareaandhighphotocatalyticactivity ,becausetheyhavegreatpotentialforsuchapplicationsasenvironmentalpurification ,decompositionofcarbonicacidgas,andgenerationofhydrogengas .Titaniananot…     

Ti(IV)与铜离子共掺杂的羟基磷灰石薄膜弱紫外光杀菌研究

采用共沉淀水热合成和离子交换的方法,制备了钛与铜离子共掺杂的羟基磷灰石(Ca10(PO4)6(OH)2:HAP)(HAPTiCu),以大肠杆菌为目标物,研究了不同材料负载的多孔镍网薄膜在弱紫外光下的杀菌活性.结果表明,HAPTiCu在弱紫外光下具有高效杀菌效率,并远远高于HAPTi和P25TiO2薄膜.这主要是铜离子的抗菌与光催化分解细菌的协同作用的结果.电子自旋共振(ESR)研究表明杀菌反应的主要活性物种是O-2·.HAPTiCu薄膜释放的铜离子在光催化杀菌反应过程中,被光催化还原沉积在表面,导致了较低的铜离子溶出,从而保证了材料的稳定性.     

纳豆激酶原核表达载体的构建及其活性鉴定

利用引物搭桥的方法,经PCR扩增,获得了纳豆激酶成熟肽基因(NK),从而构建了大肠杆菌表达质粒NK/pTWIN1,NK/PET32a及NK/PML-c2x,经分别转化宿主菌ER2566,BL21和ER2566,获得了转纳豆激酶基因重组菌.结果表明,NK/pTWIN1-ER2566和NK/PET32a-BL21的纳豆激酶蛋白表达量较高.本研究选用NK/pTWIN1-ER2566作为表达菌株,对其进行发酵表达.SDS-PAGE显示,目的蛋白约占菌体总蛋白的36%,该蛋白是以包涵体形式存在的.包涵体经收集、蛋白变性及复性,并经过SP Sepharose分离纯化,得到了纯化的纳豆激酶蛋白,琼脂糖-纤维蛋白平板法测出1mg纳豆激酶干粉的溶栓活性相当于600u尿激酶.本文从基因工程角度研究纳豆激酶基因的克隆、表达及纯化,为利用基因工程菌生产纳豆激酶奠定了基础.图5表1参11     

有机溶剂,表面活性剂及NaCl对对二氯苯在水中活度系数的影响

本文利用气相色谱法测定了对二氯苯在不同组成的水溶液体系中的活度系数,其中包括二组份与三组份共溶剂体系、表面活性剂-水体系与NaCl-水体系。实验结果表明:对二氯苯在二组份和三组份共溶剂体系中的活度系数的对数值与各组份的体积分数之间有线性加和关系;表面活性剂在达到临界胶束浓度以上时,可显著降低对二氯苯在水中的活度系数;对二氯苯在NaCl水溶液中的活度系数与NaCI的含量之间成正比关系。作者还利用UNIFAC法估算了对二氯苯在各种共溶剂体系中的活度系数,所得估算值与测定值相接近。     

MOVEMENT BEHAVIOUR OF MEDAKA (Oryzias latipes) IN RESPONSE TO SUBLETHAL TREATMENTS OF DIAZINON AND CHOLINESTERASE ACTIVITY IN SEMI-NATURAL CONDITIONS

Behavioural changes of medaka (Oryzias latipes) treated with an anticholinesterase insecticide, diazinon (0.1 mg L^{- 1}), were continuously observed for 4 days in semi-natural conditions. Although variations occurred in individual specimens, the movement tracks appeared differently with typical short-range movement with irregular turns and shaking after the treatments. Eight movement patterns frequently observed before and after the treatments were selected, and the variables characterising the movement patterns were compared quantitatively. The variables were clearly differentiated when the movement patterns were correspondingly matched before and after the treatments (e.g., vertical movements, horizontal movements, etc). Meander and stop duration were highly different among the selected movement patterns. Additionally, different degree of toxic response behaviours could also be detected by quantitative characterisation of the variables. Response behaviour was confirmed with toxicological experiments that show the decrease in the acetylcholine esterase activity in the head and body of specimens. Quantitative investigations on the variables of the movement tracks suggested the usefulness of response behaviour as a monitoring tool for environmental assessment.     

Water Environmental Degradation of the Heihe River Basin in Arid Northwestern China

Water environmental degradation is a major issue in the Heihe River Basin belonging to the inland river basin of temperate arid zone in northwestern China. Mankind’s activities, such as dense population and heavy dependence on irrigated agriculture, place immense pressure on available and limited water resources during the last century, especially the recent five decades. An investigation on the water environmental degradation in the Heihe River Basin and analysis of its causation were conducted. The results indicated that water environmental changes in the whole basin were tremendous mostly in the middle reaches, which reflected in surface water runoff change, decline of groundwater table and degeneration of surface water and groundwater quality. Some new forms of management based on traditional and scientific knowledge must be introduced to solve problems of water environmental degradation in the Heihe River Basin.     

不同种类食细菌线虫对石油污染土壤微生物的影响

通过在石油污染土壤中接种不同种类食细菌线虫,探究土壤微生物活性和多样性的变化.本试验设置5个处理：杀灭线虫土壤（NFS）、石油污染土壤（SP）、在石油污染土壤中分别接种不同线虫,即秀丽隐杆线虫（Caenorhabditis elegans,SPN1）、头叶属线虫（Cephalobus persegnis,SPN2）和小杆属线虫（Rhabditis marina,SPN3）.结果表明,经过168d试验,和NFS相比,SP的土壤基础呼吸和脲酶活性增大,而微生物量碳含量和蔗糖酶活性减小.和SP相比,SPN1、SPN2、SPN3的基础呼吸增大,微生物量碳减少了34.59%~72.48%,蔗糖酶活性增加了15.66%~22.89%.SP、SPN1、SPN2和SPN3的微生物的丰富度（S）分别下降了5.00,7.25,2.50和9.75,Shannon-Wiener指数（H'）分别显著下降了0.18,0.15,0.15和0.23.不同种类的食细菌线虫可通过捕食作用对石油污染土壤微生物活性及多样性产生影响并有种间差异,有利于提高石油污染土壤微生物活性,促进石油降解.     

有机负荷和温度波动对厌氧菌群及酶活影响

基于太阳能辅热厌氧消化反应器进行餐厨垃圾半连续发酵试验,探究了温度波动和有机负荷调控（OLR=2.0、4.0、6.0、7.0 kg·m^-3·d^-1）对甲烷产量、酶活性变化和微生物群落结构的影响.结果表明,反应器可以在OLR为2.0 kg·m^-3·d^-1下稳定运行并在6.0 kg·m^-3·d^-1时实现最佳甲烷生产效率,虽然太阳能组比电能组减少了4倍能耗,但热辐射不稳定导致太阳能组发酵温度波动,甲烷平均产量比电能组减少21%.此外,蛋白酶在温度波动环境下表现出较高活性,但脂肪酶和淀粉酶活性却受到抑制.高通测序结果表明,低OLR阶段乙酸型产甲烷菌Methanosaeta活性较强,随着OLR递增氢营养型产甲烷菌Methanoregula和Methanospirillum相对丰度逐渐提高,而试验全过程中水解细菌Firmicutes相对丰度维持在62%~95%,占据主导地位.