Oxidative damage in unfertilized eggs of Chinese rare minnow (Gobiocypris rarus) exposed to nonylphenol

In the present study, female Chinese rare minnows (Gobiocypris rarus) were used as in vivo models and exposed to nonylphenol (NP) at concentrations of 1 to 200 mu g/L for 21 d under semistatic conditions. Molecular biomarkers of oxidative stress were measured in unfertilized eggs and included reactive oxygen species (ROS), lipid peroxidation products (thiobarbituric acid-reactive substances [TBARS] and protein carbonyl), superoxide dismutase activity, and glutathione. Cathepsin D activity as an indicator of egg viability also was assayed. Nonylphenol induced ROS formation in unfertilized eggs in all exposed groups compared to the controls. The levels of protein carbonyl and TBARS in unfertilized eggs were significantly increased (p < 0.05) at 10 to 200 and 100 to 200 mu g/L, respectively. Good positive correlations were shown between ROS induction and levels of TBARS and protein carbonyl in eggs (R = 0.918, p < 0.05 and R = 0.784, p < 0.05, respectively). Superoxide dismutase activity in eggs was significantly inhibited (p < 0.05) in the 50 to 200 mu g/L exposure groups. Glutathione levels in eggs were significantly depleted (p < 0.05) at 100 to 200 mu g/L concentrations. In addition, ROS induction resulted in oxidative damage to lipid and protein in chorions. Significant reductions (p < 0.05) of the protein and lipid contents in chorions were both found in the 50 to 200 mu g/L exposure groups. A previous study found that NP exposure could lead to chorion thinning in zebra fish. Thus, the reductions in protein and lipid contents in chorion could be the reason for chorion thinning by NP exposure. Meanwhile, cathepsin D activity was significantly inhibited (p < 0.05) in all exposure groups. The results demonstrated that NP-induced oxidative stress could damage the chorion of unfertilized eggs and lead to a decline in gamete quality in female Chinese rare minnow.

Nonylphenol and octylphenol in urban eutrophic lakes of the subtropical China

The paper studied two estrogenic pollutants, 4-nonylphenol (NP) and 4-tert-octylphenol (OP) in water, suspended particle (SP) and sediments in urban eutrophic lakes. The concentrations of NP ranged from 1.94 to 32.85 mu g/l, 0.876 to 31.13 mu g/l and 3.54 to 32.43 mu g/g dry weight (dw) in water, suspended particle (SP) and sediments, respectively, and that of OP from 0.027 to 1.44 mu g/l, 0.008 to 1.777 mu g/l and 0.058 to 1.245 mu g/g dw in water, suspended particle (SP) and sediments, respectively. An increasing trend in the concentration was noticed in all matrices close to the sewage inlets, which was found to be the major factor affecting the spatial distribution of alkylphenols (APs) in the lakes. Due to restoration of submerged macrophytes, which might accumulate APs, the contaminations of APs in the Little Moon Lake (LML) and the Little Lotus Lake (LLL) were lower than those in the Big, Moon Lake (BML) and the Bier Lotus Lake (BLL). A reasonable correlation of NP and OP was obtained among water, suspended particle and sediment. The possible environmental stress of APs concentration on aquatic organisms in Wuhan urban lakes was also discussed.

Effects of nonylphenol on the growth and microcystin production of Microcystis strains

Both organic pollution and eutrophication are prominent environmental issues concerning water pollution in the world. It is important to reveal the effects of organic pollutants on algal growth and toxin production for assessing ecological risk of organic pollution. Since nonylphenol (NP) is a kind of persistent organic pollutant with endocrine disruptive effect which exists ubiquitously in environments, NP was selected as test compound in our study to study the relationship between NP stress and Microcystis growth and microcystin production. Our study showed that responses of toxic and nontoxic Microcystis aeruginosa to NP stress were obviously different. The growth inhibition test with NP on M. aeruginosa yielded effect concentrations EbC50 values within this range of 0.67-2.96 mg/L. The nontoxic M. aeruginosa strains were more resistant to NP than toxic strains at concentration above 1 mg/L. Cell growth was enhanced by 0.02-0.2 mg/L NP for both toxic and nontoxic strains, suggesting a hormesis effect of NP on M. aeruginosa. Both toxic and nontoxic strains tended to be smaller with increasing NP. But with the increased duration of the experiment, both the cell size and the growth rate began to resume, suggesting a quick adaptation of M. aeruginosa to adverse stress. NP of 0.05-0.5 mg/L significantly promoted microcystin production of toxic strain PCC7820, suggesting that NP might affect microcystin production of some toxic M. aeruginosa in the field. Our study showed that microcystin excretion was species specific that up to 75% of microcystins in PCC7820 were released into solution, whereas > 99% of microcystins in 562 remained in algal cells after 12 days' incubation. NP also significantly influenced microcystin release into cultural media. The fact that NP enhanced growth and toxin production of M. aeruginosa at low concentrations of 0.02-0.5 mg/L that might be possibly found in natural freshwaters implies that low concentration of NP may favor survival of M. aeruginosa in the field and may play a subtle role in affecting cyanobacterial blooms and microcystin production in natural waters. (c) 2006 Elsevier Inc. All rights reserved.

Reproductive effects of prenatal exposure to nonylphenol on zebrafish (Danio rerio)

Mature female and male zebrafish were separated and exposed to nonylphenol (NP) at 0.1, 1, 10, 50, 100 and 500 mu g/L, respectively, for 3 weeks. Gonadosomatic index (GSI) in both sexes and vitellogenin (VTG) induction in males was measured as the bioindicators for the impairment to the parents. The results indicated that 50 mu g/L of NP was the non-observed effect concentration (NOEC) for GSI and VTG induction. Afterwards, the 50 mu g/L NP exposed females and males, and the control females and males were cross-wise pair-bred in the control water for one week to examine the reproductive effects. The embryonic cathepsin D (CAT D) activity, eggshell thickness, fecundity, hatching rate and malformation (vertebral column flexure) rate of offspring were determined in the four pair-bred groups. While endpoints remained unchanged in the groups with exposed males, prenatal exposure of females to 50 mu g/L of NP resulted in the impairment of reproduction in groups with exposed females including inhibition of CAT D activity (P < 0.05), decrease of eggshell thickness (by 23.6%) and elevation of malformation rate (P < 0.001). These results suggested NP could induce reproductive damage to zebrafish at NOEC for parents. The results also imply that alterations of CAT D activity and eggshell thickness may be more sensitive biomarkers to indicate the reproductive effects caused by endocrine disrupting chemicals. (c) 2005 Elsevier Inc. All rights is reserved.

Nonylphenol, bisphenol-A and DDTs in Lake Donghu, China

A preliminary survey was carried out in April 2003 to estimate the levels of nonylphenol (NP), bisphenol-A (BPA) and residual DDTs in Lake Donghu. After then, the sediments of some areas of the lake were moved out for clearing the lake by the local government. Therefore, the variance of NP and BPA after the clearance of sediment in the surface water was determined from December 2003 to May 2004. Sediments, surface water and fish were collected from four sub-lake areas and the analytes were qualified by GC/MS in SIM mode after concentration onto an Oasis solid phase extraction cartridge. NP and BPA values ranged between 5.46-119.10 and 0.9913.42 mg/kg dw, respectively, in sediments, 75.2-179.6 and 15.1-62.5 mu g/L in surface water. Meanwhile, the bioaccumulation factors (BCFs) of NP and BPA were calculated in fish livers. Plasma vitellogenin (VTG), a sensitive biomarker showing oestrogenicity was detected in the captured male Wuchang bream (Megalobrama amblycephala Yih) and common carp (Cyprinus carpio). After sediment clearance, the concentrations of NP and BPA decreased to 0.65-25.04 and 0.04-21.32 mu g/L. The results indicate the presence of high-dose xenoestrogenic NP and BPA, probably major contributors which associate with VTG induction in Lake Donghu male fish.

The accumulation of alkylphenols in submersed plants in spring in urban lake, China

The concentrations of alkylphenols (APs) were investigated in water, sediments and submersed macrophytes from the Moon Lake, Wuhan city, China. The water samples contained APs, ranging up to 26.4 mu g l(-1) for nonylphenol (NP) and 0.68 mu g l(-1) for octylphenol (OP). APs were found in the sediment samples with concentrations ranging from 4.08 to 14.8 for NP and from 0.22 to 1.25 mu g l(-1) dry weight for OP. The samples from the site near former sewage inlet showed the highest concentrations of APs in both water and sediments. The results of distribution pattern and dynamics of NP and OP in submersed macrophytes of the Moon Lake showed that the two pollutants were all found in Myriophyllum verticillatum, Elodea nuttallii, Ceratophyllum oryzetorum, and Potamageton crispus collected from the Moon Lake. For NP, M. verticillatum had the highest capacity of accumulation, followed by E. nuttallii, C. oryzetorum and P. crispus. However the distribution pattern of OP differed from that of NP, and the highest amount of accumulation was observed in E. nuttallii, followed by M. verticillatum, P. crispus, and C. oryzetorum. The temporal pattern of APs was also observed in submersed macrophytes from March to May, and the highest accumulation period was in May. (c) 2008 Elsevier Ltd. All rights reserved.

Seasonal variations of estrogenic compounds and their estrogenicities in influent and effluent from a municipal sewage treatment plant in China

The seasonal variations of estrogenic compounds and the estrogenicities of influent and effluent were investigated by OF chemical analysis and in vitro assay in a municipal sewage treatment plant in Wuhan (China). The levels of eight estrogenic compounds, including 17 beta-estradiol (E-2) estrone (E-1), estriol (E-3) diethylstilbestrol (DES), 17 alpha-ethinylestradiol, nonylphenol (NP), 4-tert-octylphenol (OP), and bisphenol A (BPA), were measured by gas chromatography-mass spectrometry. Total estrogenic activity of sewage was quantitatively assessed using primary cultured hepatocytes of male Megalobrama amblycephala Yih using vitellogenin as a biomarker. The E-2 equivalents (EEQs) obtained from the chemical analysis were consistent with those measured by bioassay. The natural (E-1, E-2, and E-3) and synthetic (DES) estrogens, as well as NP, were the main contributors of the total EEQs of influent and effluent in the present study. The levels of natural estrogens E-1 and E-3 in the influent and effluent were higher in winter than in summer, whereas the situation for NP and OP was the reverse. The levels of E-2, DES, and BPA varied little among different seasons. 17 alpha-Ethinylestradiol was not detected in the influent and effluent. The estrogenicities of the influent and of the primary and secondary effluents were all higher in summer than in winter. Estrogenic activities in winter mainly originated from natural (E-1, E-2, and E-3) and synthetic (DES) estrogens, whereas the increase of EEQs in summer was contributed by NP The results from chemical analysis and bioassay demonstrate that estrogenic compounds cannot be entirely removed by the existing sewage treatment process, which should be further improved to protect aquatic ecosystems and human health.

Evaluation of estrogenic activities and mechanism of action of perfluorinated chemicals determined by vitellogenin induction in primary cultured tilapia hepatocytes

Perfluorochemicals (PFCs) are emerging persistent organic pollutants (POPs) and are widely present in the environment, wildlife and humans. Recently, reports have suggested that PFCs may have endocrine-disrupting activities. In the present study, we have developed a non-competitive enzyme-linked immunosorbent assay (ELISA) method to investigate estrogenic activities of selected PFCs using vitellogenin (VTG) induction in primary cultured hepatocytes of freshwater male tilapia (Oreochromis niloticus). Cultured hepatocytes were exposed to various concentrations of perfluorooctanyl sulfonate (PFOS), pentadecafluorooctanoic acid (PFOA), 1H, 1H, 2H, 2H-nonafluoro-1-hexanol (4:2 FTOH), 1H, 1H, 2H, 2H-perfluorooctanol (6:2 FTOH) and 1H, 1H, 2H, 2H-perfluoro-1-decanol (8:2 FTOH) for 48h, while 17 beta-estradiol (E2) and 4-nonylphenol (4-NP) were used as positive controls. A dose-dependent induction of VTG was observed in E2-, 4-NP-, PFOS-, PFOA- and 6:2 FrOH-treated cells, whereas VTG levels remained unchanged in the 4:2 FTOH and 8:2 FTOH exposure groups at the concentrations tested. The estimated 48-h EC50 values for E2,4-NP, PFOS, PFOA and 6:2 FTOH were 4.7 x 10(-7), 7.1 x 10(-6), 1.5 x 10(-5), 2.9 x 10(-5) and 2.8 x 10(-5) M, respectively. In the time-course study, significant VTG induction took place at 24 h (E2), 6 It (4-NP), 48 It (PFOS), 48 It (PFOA), 72 It (4:2 FTOH), 12 h (6:2 FTOH), 72 h (8:2 FTOH), and increased further after 96 It of exposure. Co-exposure to binary mixtures of individual PFCs and E2 for 48 It significantly inhibited E2-induced hepatocellular VTG production in a dose-dependent manner except for 4:2 FTOH. The estimated 48-h IC50 (concentration of a compound that elicits 50% inhibition of maximally E2-induced VTG) values for PFOS, PFOA, 6:2 FTOH and 8:2 FTOH were 3.1 x 10(-7), 5.1 X 10(-7), 1.1 X 10(-6) and 7.5 x 10(-7) M, respectively. In order to further investigate the estrogenic mechanism of PFCs, the hepatocytes were co-exposed to binary mixtures of individual chemicals (E2,4-NP, PFOS, PFOA and 6:2 FTOH) and the known estrogen receptor inhibitor tamoxifen for 48 h; tamoxifen significantly inhibited the ability of these chemicals to stimulate vitellogenesis. The overall results demonstrated that PFOS, PFOA and FTOHs have estrogenic activities and that exposure to a combination of E2 and PFCs produced anti-estrogenic effects. The results of the estrogen receptor inhibition assay further suggested that the estrogenic effect of PFCs may be mediated by the estrogen receptor pathway in primary cultured tilapia hepatocytes. (c) 2007 Elsevier B.V. All rights reserved.

NP难问题的算法和实验性研究

NP难是计算机科学中的一个重要概念和核心问题，自从它的提出到现在， 人们已经得到了很多重要的理论结果。直观上讲，一个问题一旦被证明是NP难 的就意味着我们很难找到该问题的一个多项式时间的有效算法。但从实用的角 度讲，对于应用中遇到的问题，单单是证明它很难（是NP难的）是不够的，如何 在合理的时间内求解实际问题也是必须解决的现实问题。本文主要侧重于NP难 问题的算法和实验性研究，研究对象主要是可满足性问题、图的顶点染色、图 的子图匹配等NP难问题，以及可满足性模理论的解空间计算和体积估算等扩展 问题。 围绕几个著名的问题，本文的主要工作如下： 针对图染色问题，日本研究人员提出了一种通过组合小图单元得到大的难 实例的方法。他们通过试错的方式手工找到了7个小图单元。我们提出了一种新 的构造算法来系统地生成这类小图单元，用我们的算法生成的难图染色实例， 主流的图染色工具需要指数时间才能求解；在一些专门求解色数比较小的图的 图染色工具上我们的算法生成的实例更难求解。针对皇后图染色问题，我们利 用模型查找工具SEM来对这类问题进行求解，在求解过程中提出了新的变量选 择策略，发现比简单地使用可满足性问题工具和图染色工具效果要好。 针对语义Web推理中的关键问题RDF蕴含关系的判定问题，我们利用从子 图同构问题到可满足性问题的编码方案，把它转化为命题逻辑公式的可满足性 判定问题，并采用了启发式的方法对编码过程进行必要的化简得到较少的布尔 公式，然后再利用高效的可满足性问题工具来求解。这种转化为可满足性问题 的方法，是跟RDF简单蕴含的模型论语义结合比较自然的一个方法。在小规模 实例上，这种方法的效果也很好。 针对布尔和数值混合约束的公式，即可满足性模理论（线性理论）公式的 体积计算这一新问题，我们首先给出一个直接计算体积的方法，然后提出一个 改进的算法，并研究了如何通过引入可满足性模理论中的技术来尝试对该算法 进一步地改进。我们实现了工具并做了实验。在一个实际的程序实例上，我们 还就“热门路径”问题做了实例研究和探讨。 体积计算是一个有广泛应用背景的经典难题（#P难的），但以前的方法要 么只能处理线性约束，要么只具有理论价值（不够实用）；针对含非线性约束的 体积计算问题我们提出了实用的算法，并设计了相应的工具，在低维实例有很 好的逼近效果。

纯公钥模型下对NP语言的高效并发零知识证明系统

提出了一种从3轮公开掷币的对任何NP语言的诚实验证者零知识证明系统到纯公钥模型下4轮f轮最优）对同一语言的具有并发合理性的并发零知识证明系统．该转化方法有如下优点：1）它只引起D（1）（常数个）额外的模指数运算，相比DiCrescenzo等人在ICALP05上提出的需要qn）个额外的模指数运算的转化方法孩系统在效率上有着本质上的提高，而所需的困难性假设不变；2）在离散对数假设下，该转化方法产生一个完美零知识证明系统．注意到DiCrescenzo等人提出的系统只具有计算零知识性质．该转化方法依赖于一个特殊的对承诺中的离散对数的3轮诚实验证者零知识的证明系统．构造了两个基于不同承诺方案的只需要常数个模指数运算的系统这种系统可能有着独立价值．