A preliminary study of temporal differences in serum concentrations of perfluoroalkyl acids,among New York anglers,in the absence of known changes in manufacturing practices

Perfluorinated compounds have been manufactured in large quantities and used in myriad industrial processes and commercial applications. The aim of this preliminary study was to generate hypotheses with regard to differences in body burdens of perfluoroalkyl acids, among a sub-sample of participants from the New York State Angler Cohort Study, over a time interval during which no known substantial changes occurred in US manufacturing practices or commercial use. Paired serum specimens, collected from 15 subjects in 1993–1994 (time 1), and in 1995–1997 (time 2), with a minimum interval of 2.5 years, were assayed for PFDA, PFHpA, PFHxS, PFNA, PFOA, PFOS, PFOSA and PFUnDA using HPLC with ES-MS/MS. By subject, differences in concentrations between time 1 and time 2 were estimated, employing paired t-tests, correlations, and multivariable linear regression to accommodate heterogeneity in duration between specimens, and in time 1 concentrations. A statistically significant (P < 0.05) adjusted mean decrease of 0.16 ng mL^?1 (18.8%) between time 1 and 2 was detected for PFNA, and an adjusted mean increase of 0.54 ng mL^?1 (56.8%) was detected for PFOA. The results of this study may be indicative of short-term changes in human body burdens of PFNA and PFOA in association with local exposure sources.     

Lack of genotoxic potential of permethrin in mice evaluated by the comet assay and micronucleus test

Permethrin is a common insecticide that does not show genotoxic potential in standard in vitro and in vivo assays. To investigate the genotoxic potential of permethrin in more detail, two in vivo studies were conducted on female mice to assess DNA damage in tumor target organs by the comet assay and micronucleus test. For this, mice were administered permethrin at doses of 150, 300, or 600 mg/kg/day by gavage for 2 days, and their lung, liver, glandular stomach, peripheral blood, and bone marrow cells were examined for DNA damage. There were no significant increases in % tail DNA in the organs examined and no increase in micronuclei in peripheral blood by flow cytometry. Taken together, the present findings provide evidence that permethrin has no genotoxic, aneugenic, or clastogenic potential.     

Induction of micronuclei in bone marrow cells and hepatotoxicity of one of the most common over-the-counter pyrethroid insecticide products in Nigeria

Increasing environmental pollution in the tropics is creating a breeding ground for mosquitoes, leading to increasingly frequent use of insecticides to combat home infestation of mosquitoes, the vector of malarial parasites. Household members are therefore more prone to aerosol exposure and subsequent health effects. We assessed the hepatotoxic and clastogenic effects in rats exposed to different levels of aerosols of one of the insecticides most commonly in used in Nigeria. There were significant (p?2 in the groups of rats exposed to the aerosols when compared with the control group. In addition, rats exposed to the insecticide aerosols have elevated activities of serum enzymes: γ-glutamyl transferase, aspartate amino transferase and alanine amino transferase, as well as number of micronuclei scored in the bone marrow, at levels that are significantly (p?相似文献   

六溴环十二烷(HBCD)对H4细胞和SK-N-AS细胞脱碘酶2和3表达的影响

六溴环十二烷(hexabromocyclododecane,HBCD)与多溴联苯醚(polybrominated diethyl ethers,PBDEs)复合污染体系,对人类健康尤其神经系统所造成的潜在危害及其机制一直是笔者课题组的研究方向。HBCD是广泛使用的溴化阻燃剂,与PBDEs一样,会通过干扰内分泌系统、影响甲状腺激素分泌及损伤神经系统,对生物体产生发育神经毒性。作为系列研究之一,本研究以H4人脑神经胶质瘤细胞和SK-N-AS人神经母细胞瘤细胞为体外生物模型,通过观察HBCD对H4细胞Ⅱ型脱碘酶(Dio2)和SK-N-AS细胞Ⅲ型脱碘酶(Dio3)表达的调控,初步探讨了HBCD对神经系统局部甲状腺激素水平的潜在影响。H4细胞和SK-N-AS细胞分别暴露于0、1、3和9μmol·L-1HBCD 24 h后,采用MTT法检测细胞活力,Western Blot和RT-PCR法分别分析Dio2和Dio3蛋白和基因的表达,酶联免疫吸附测定(ELISA)法检测H4细胞脑源性神经细胞营养因子(BDNF)的分泌。结果表明,HBCD以剂量依赖方式降低H4细胞和SK-N-AS细胞生存率,引起H4细胞Dio2蛋白和基因表达下调,而致SK-N-AS细胞Dio3蛋白和基因表达上调。此外,HBCD还降低H4细胞BDNF的分泌。这表明,HBCD很可能通过影响神经和胶质细胞脱碘酶的表达,影响脑局部甲状腺激素水平,从而引起神经系统损伤及发育神经毒性。     

邻苯二甲酸二异辛酯致赤子爱胜蚓体细胞氧化损伤的研究

以赤子爱胜蚓为实验材料,探讨了邻苯二甲酸二异辛酯(DEHP)对赤子爱胜蚓(Esisenia foelide)的毒性.DEHP染毒剂量:0、0.2、0.4、0.8、1.6mmol·L-1,染毒2h后采用KCl-SDS沉淀法检测细胞匀浆DNA-蛋白质交联(DPC)系数,硫代巴比妥酸法检测丙二醛(MDA)含量.结果表明:随着DEHP染毒浓度的增加,赤子爱胜蚓细胞匀浆的DPC系数显著升高(F=14.1,p<0.01),并具有剂量-效应关系(r=0.8877,p<0.05),而MDA含量变化不显著,各染毒组与对照组之间不存在显著性差异(F=0.27,p>0.05).以上结果表明,实验浓度的DEHP暴露可使赤子爱胜蚓体细胞发生显著的DNA蛋白质交联,具有遗传毒性.     

Isolation and identification of fungi tolerant to polycyclic aromatic hydrocarbons and coal tar from different habitats in Lithuania

A group of 36 fungal strains, belonging to the Lithuanian mycobiota, was collected and isolated from different locations, habitats, and matrices, including creosote-treated wood in storage yards for crosstie wastes. The eight most perspective strains selected according to preliminary assessment of tolerance to coal tar were subsequently identified combining taxonomical evaluation and molecular techniques. The tolerance of the eight identified fungal species (five basidiomycetes and three ascomycetes) to the presence of various concentrations of coal tar, and for the four most perspective fungal strains (Pleurotus sp., Schizophyllum sp., Irpex lacteus, Bjerkandera adusta) to polycyclic aromatic hydrocarbons was evaluated. The ligninolytic enzymatic activity assay of the isolated strains resulted in a good correspondence between the tolerance to pollutants and the capability to produce ligninolytic enzymes indicating that this group of white-rot fungi is perspective for further investigation and eventual usage for mycoremediation of polycyclic aromatic hydrocarbons polluted substrates.     

Genotoxicity assessment of human peripheral Lymphocytes induced by thallium(I) and thallium(III)

Thallium is a non-essential metal with a wide range of industrial uses. However, thallium is also a potential pollutant with high potential toxicity to humans. In the present study, we analyzed and compared the cellular and genotoxic effects of thallium in two main oxidation states by applying chromosome aberration assays to human peripheral lymphocytes. We observed that thallium(I) sulfate reduced the mitotic index at all tested concentrations (0.5, 1, 5, 50 and 100 μg/mL), whereas thallium(III) chloride was toxic at concentrations ≥1 μg/mL. Thallium(I) and thallium(III) treatment significantly increased structural chromosomal aberrations, with and without gaps, and increased the percentage of aberrant cells without gaps. Furthermore, satellite associations and numerical chromosomal aberration tests showed significant differences at a few of the tested concentrations. The satellite association test is related to aneuploidy. Thallium salts increased satellite associations when hyperploid cells were observed. Our results indicated that the two oxidation states of thallium induced toxicity in vitro – i.e. cyto/genotoxic (clastogenic and aneuploidogenic) effects.     

磷酸三苯酯(TPP)和磷酸三(2-氯异丙基)酯(TCPP)诱导人胚肾细胞HEK293凋亡的机制研究

磷系阻燃剂对人体的潜在毒性作用引起了国内外研究者的广泛关注。肾脏是机体重要的排毒器官,若肾脏细胞受损,可能影响肾脏功能的正常发挥。本研究以人胚肾细胞HEK293为研究对象,结合传统毒理学实验,筛选出磷酸三苯酯(TPP)及磷酸三(2-氯异丙基)酯(TCPP)诱导人胚肾细胞HEK293凋亡的关键靶标基因p53。在此基础上采用分子对接模拟和光谱法分析发现,TPP和TCPP分别以嵌插方式和沟槽方式结合p53-DNA,改变基因片段的框架结构,启动分子起始事件,通过影响相关基因(Bax、Hrk、Bcl-2和Bad)的表达量,导致线粒体途径释放cyt c,最终激活Caspase 7实现细胞凋亡。研究结果阐明了此类污染物诱导凋亡的作用机制,为毒害化学品的污染防控提供理论依据。     

太阳鱼脑乙酰胆碱酯酶的分离纯化

采用硫酸铵沉淀–普鲁卡因胺亲和柱层析方法,对太阳鱼(Lepomis gibbosus)脑部乙酰胆碱酯酶分离纯化,纯化倍数为234倍,回收率为25.67%,经SDS-PAGE电泳检测,呈现单一的条带,相对分子质量约为68 000,表明其纯度达到电泳纯.     

Distribution-free two-sample comparisons in the case of heterogeneous variances

Behavioral ecologists are often faced with a situation where they need to compare the central tendencies of two samples. The standard tools of the t test and Mann–Whitney U test (equivalent to the Wilcoxon rank-sum test) are unreliable when the variances of the groups are different. The problem is particularly severe when sample sizes are different between groups. The unequal-variance t test (Welch test) may not be suitable for nonnormal data. Here, we propose the use of Brunner and Munzel’s generalized Wilcoxon test followed by randomization to allow for small sample sizes. This tests whether the probability of an individual from one population being bigger than an individual from the other deviates from random expectation. This probability may sometimes be a more clear and informative measure of difference between the groups than a difference in more commonly used measures of central tendency (such as the mean). We provide a recipe for carrying out a statistical test of the null hypothesis that this probability is 50% and demonstrate the effectiveness of this technique for sample sizes typical in behavioral ecology. Although the test is not available in any commercial software package, it is relatively straightforward to implement for anyone with some programming ability. Furthermore, implementations in R and SAS are freely available on the internet.     

邻苯二甲酸丁基苄酯致神经细胞氧化损伤

为探究邻苯二甲酸丁基苄酯(butyl benzyl phthalate,BBP)对小鼠神经的毒性作用,进行了小鼠体外毒理学研究。首先用不同浓度的邻苯二甲酸丁基苄酯染毒神经模型细胞—N2a神经瘤细胞,通过噻唑蓝比色法(MTT),Hoechst 33258染色实验评价邻苯二甲酸丁基苄酯的细胞毒效应;通过对染毒细胞氧自由基(ROS)、丙二醛(MDA)、还原型谷胱甘肽(GSH)含量的检测来探究BBP对小鼠神经瘤细胞的氧化损伤效应。随着BBP浓度的不断增高,细胞的MTT值逐渐变小,当BBP的浓度达到10 g·L-1时,MTT实验结果与对照组出现显著性差异;Hoechst 33258染色结果显示:高浓度的BBP导致细胞核呈现出不规则状态,出现了凋亡小体;随着BBP染毒浓度的升高N2a细胞中的ROS水平和MDA含量逐渐上升,分别在0.16 g·L-1和10 g·L-1开始与对照组相比出现了显著性的差异(p0.05);而GSH系数呈现下降趋势,在0.32 g·L-1时开始出现显著性差异(p0.05)。实验结果表明高浓度的邻苯二甲酸丁基苄酯可以导致神经瘤细胞的凋亡,并产生氧化损伤效应。     

Modeling toxic endpoints for improving human health risk assessment of polycyclic aromatic hydrocarbons – parent compounds and simple mixtures

Risk assessments for mixtures of polycyclic aromatic hydrocarbons (PAH) are problematic due to the lack of available potency and toxicity data on individual compounds and mixtures. This article examines the toxicity of parent compounds and designed mixtures of PAH in order to bridge the gap between component assessment and mixture assessment for this class of ubiquitous compounds. The objective for this study was to test seven parent PAH compounds and four PAH mixtures in a set of three bioassays to evaluate the toxicity of parent compound PAH and binary mixtures of PAH. PAH and mixtures were examined in the Salmonella/microsome mutagenicity assay, a Gap Junction Intercellular Communication assay, and the 7-ethoxyresorufin-O-deethylase assay. These assays were chosen for their ability to measure specific toxic endpoints related to the carcinogenic process (i.e. initiation, promotion, and progression). Two compounds similar in structure, benzo(a) pyrene (BAP) and benzanthracene, consistently produced positive results in all three bioassays. Conversely, a linear PAH, anthracene, produced negative results in all three bioassays. An antagonistic response was observed for the mixtures in all three bioassays. Chemical structure was important in explaining the observed responses. Using chemical structure–activity relationships with the steps of the carcinogenic process may be used to improve estimates of toxicity for compounds and mixtures for human health risk assessments.     

石油烃对栉孔扇贝血淋巴细胞DNA损伤的初步研究

为研究石油烃对海洋生物的毒性效应,将栉孔扇贝(Chlamys farreri)暴露于0.08、0.21和0.88mg·L-1石油烃中,采用单细胞凝胶电泳实验(彗星实验)技术检测不同暴露时间扇贝血淋巴细胞的DNA损伤程度,对照组中石油烃背景浓度为0.04mg·L-1。结果显示,低浓度(0.08mg·L-1)的石油烃短期(<7d)内即可导致栉孔扇贝血淋巴细胞的DNA损伤,并且随石油烃浓度的增大和暴露时间的延长,DNA损伤程度增加,石油烃浓度达0.88mg·L-1时,DNA损伤程度已非常严重。3d恢复实验后,各浓度组DNA损伤又均有不同程度的恢复。研究表明,彗星实验是检测石油烃对海洋贝类DNA损伤的一种有效手段,贝类血淋巴细胞DNA损伤有望成为石油烃污染的一种生物标志物,用于海洋污染的早期预警监测。     

双酚A对小鼠肝脏和肾脏细胞的氧化损伤

为探究双酚A(BPA)的氧化毒性,分别以剂量为20、40和80mg·kg~(-1)·d~(-1)的BPA对雄性昆明小鼠灌胃处理1周,并测定了小鼠体内活性氧自由基(ROS)水平、还原型谷胱甘肽(GSH)含量、丙二醛(MDA)含量和DNA-蛋白质交联系数(DPC)。与对照组相比,各BPA暴露组小鼠肝脏和肾脏细胞中的ROS生成量、MDA含量和DPC系数均升高,而GSH含量下降(P<0.05或P<0.01)。ROS生成量、GSH含量和DPC系数均显示出剂量-效应关系。研究表明,BPA可扰乱小鼠肝脏和肾脏细胞的氧化应激平衡,诱导细胞氧化损伤。     

磺胺甲恶唑污染对颤蚓的氧化胁迫

在实验室模拟条件下分别采用动、静态生物实验方法,研究了不同浓度的磺胺甲恶唑(sulfamethoxazole,SMZ)对颤蚓(Monopyle phorus limosus)体内抗氧化防御系统的影响以及颤蚓暴露于单一浓度的SMZ后,其体内典型的抗氧化酶,超氧化物歧化酶(SOD)的活性随时间的响应。结果显示:染毒8d后,颤蚓体内SOD的活性先降低后逐渐升高。SMZ染毒浓度达到1.00mg·L-1时,SOD的活性被显著诱导(P<0.01),为对照组的1.30倍。过氧化物酶(POD)活性的变化趋势明显分为先升高、再降低2个阶段。染毒浓度达到0.25mg·L-1时,POD的活性受到最大诱导(P<0.01),约为对照组的1.40倍。各处理组过氧化氢酶(CAT)的活性均高于对照组,并且随着SMZ暴露浓度的增加,CAT活性的变化趋势是逐渐降低的,与SMZ染毒浓度间存在较好的剂量-效应关系。随着SMZ暴露时间的延长,SOD、POD和CAT的活性变化趋势类似,均为先升高后降低,并在第8天时达到最大值(P<0.01),增长率分别为32%、26%和82%。综上,SMZ能诱导颤蚓体内产生氧化胁迫,而CAT在活性氧自由基的清除中发挥主要作用。     

Callyspongia samarensis (Porifera) extracts exhibit anticancer activity and induce bleaching in Porites cylindrica (Scleractinia)

Marine sponges can produce allelopathic compounds with specific roles in the competition for benthic space. Here we demonstrate that extracts from Callyspongia samarensis (Phylum Porifera) accelerate bleaching in Porites cylindrica (Phylum Cnidaria, Order Scleractinia) and exhibit in vitro anticancer activity. A column chromatography fraction and HPLC fraction, purified from the crude methanol extract of C. samarensis, were incorporated into agar gel cubes at natural concentrations and tested on P. cylindrica corals in a laboratory assay. Statistical analysis of percent bleached area and maximum quantum yield showed that a significant difference existed between P. cylindrica nubbins that were exposed to C. samarensis extracts versus the control group. This suggests that C. samarensis contains allelopathic compounds that can cause bleaching in P. cylindrica, possibly leading to tissue necrosis and death. Furthermore, the aforementioned HPLC fraction exhibited significant growth inhibition of the HCT-116 (human colon cancer) cell line. Understanding the strategies by which sponges exert dominance over other organisms is important because it provides information about the ecological roles of sponge allelopathy and can result in the discovery of compounds with biomedical potential.     

Characterization of the liver cytochrome P450 in the marine fish Mugil cephalus and effects of some hydrocarbons on the mixed-function oxidase components

The marine fish Mugil cephalus was caught in the Bay of Marseilles (France) in 1985 and the liver microsomal cytochrome P₄₅₀ was identified at a concentration of about 0.230 nmol mg^-1 protein. Optimal assay conditions were established to measure two mixed-function cytochrome P₄₅₀-dependent oxidases: aniline hydroxylase and p-nitroanisole O-demethylase. Induction of cytochrome P₄₅₀ occurred after exposure of mullet to an aromatic hydrocarbon and a water-soluble petroleum fraction. The liver cytochrome P₄₅₀ was solubilized and purified. The monomer molecular weight of partially purified cytochrome P₄₅₀, determined by sodium dodecyl-sulfate-gel electrophoresis, was 51 000.     

A comparison of Lowry,Bradford and Smith protein assays using different protein standards and protein isolated from the marine diatom Thalassiosira pseudonana

Two spectrophotometric assays for protein commonly used in marine research (Coomassie stain, Bradford; alkaline copper, Lowry) and a more recent assay which has not been applied in this field (bicinchoninic acid, Smith) were compared for homogenates of the marine diatom Thalassiosira pseudonona using bovine serum albumin (BSA) as a standard. When homogenates were prepared by precipitating protein with trichloroacetic acid (TCA) and redissolving in 1 N NaOH, the protein content estimated by the Lowry and Smith assays agreed closely, but was consistently 20% higher than that indicated by the Bradford assay. To determine if this difference was due to the choice of a protein standard, protein from T. pseudonana was purified and compared to BSA, bovine gamma-globulin (BGG), and casein. The reactivity of the purified protein (expressed as the slope of the absorbance vs protein concentration curve) did not differ between cultures grown at high or low irradiance. For the Smith and Bradford assays the reactivity of BSA was not significantly different from algal protein, but for the Lowry assay, algal protein was significantly higher in reactivity than BSA. BGG was not significantly different in reactivity from algal protein for the Lowry and Smith assays, but BGG gave significantly lower absorbances than algal protein in the Bradford assay. These results suggest that BSA is a suitable standard for algal protein in the Bradford assays, while BGG is preferable for the Lowry assay. Either protein standard could be used for the Smith assay. Differences in purified algal protein reactivity compared to BSA could not account for the differences among the assays, nor could interference by chlorophyll a. Precipitating protein with TCA prior to analyses gave lower protein than direct analyses of homogenates for the Lowry and Smith assays, but no differences were found for the Bradford assay. As a result, the Lowry and Smith assays indicated up to 60% greater protein than the Bradford if TCA precipitation was not performed. This may be due to removal of free amino acids and small peptides which are less reactive in the Bradford assay. The 20% higher protein found in the Lowry or Smith vs Bradford assays may be due to different assay sensitivity to small peptides or other compounds which are precipitated along with proteins by TCA. Although the Smith assay is substantially simpler to perform than the Lowry, there appear to be no quantitative differences in the results. It remains unclear which spectrophotometric assay is most accurate, but the Bradford assay is faster and simpler, and is less likely to be affected by non-protein compounds found in marine phytoplankton.     

Source characterisation and mid-term spatial and temporal distribution of polycyclic aromatic hydrocarbons in molluscs along the Basque coast (northern Spain)

The variability of polycyclic aromatic hydrocarbons (PAH) measured in the soft tissues of Mediterranean mussels (Mytilus galloprovincialis) and Pacific oysters (Crassostrea gigas) are investigated. Samples were collected from estuarine waters within the Basque Country (Bay of Biscay), between 2003 and 2011. PAH bioaccumulation showed some seasonality and significant differences were observed between cold (autumn–winter) and warm (spring–summer) seasons. Sites located within the ports of Bilbao and Pasaia showed the highest PAH concentrations in molluscs, and the highest percentages of samples above the established Environmental Quality Standards and Environmental Assessment Criteria. Probably due to human activities carried out in the area, no clear trends were observed, between 2003 and 2011, for the autumn data. Since the Basque coast is an area with high population density and industrial activity, the congener profiles (which reveal the predominance of tetra-aromatics) and the diagnostic ratios identified urban/industrial combustion processes as the main PAH sources. However, natural and petrogenic sources cannot be disregarded.     

两种典型氯酚类化合物暴露致稀有鮈鲫肝细胞的损伤效应（Toxicity Effects of Two Kinds of Typical Chlorophenols on Hepatocytes of Gobiocypris Rarus）

为探讨两种典型氯酚类化合物——三氯酚(Trichlorophenol,TCP)、五氯酚(Pentachlorophenol,PCP)单独和混合暴露对稀有鮈鲫(Gobiocyprisrarus)肝脏细胞的毒性效应,实验建立了稀有鮈鲫肝细胞原代培养模型,通过噻唑蓝实验(MTT法)和单细胞凝胶电泳(SCGE)实验分别研究了TCP、PCP及其混合暴露对稀有鮈鲫肝细胞存活率的抑制作用,以及对DNA的损伤作用.结果显示:与对照组相比,TCP和PCP单独和混合暴露均对肝脏细胞的正常生长产生了抑制作用.低剂量暴露时(如0.5、2.5μg·L-1)具有显著的抑制作用(p<0.05);高剂量暴露时(如25、50μg·L-1)具有极其显著的抑制作用(p<0.01).SCGE实验结果显示:与对照组相比,暴露剂量高于12.5μg·L-1的TCP、PCP及其混合暴露均能引起稀有鮈鲫肝细胞DNA的显著损伤(p<0.05,p<0.01).以上结果提示,TCP和PCP具有肝细胞毒性,可以引起稀有鮈鲫肝细胞DNA损伤.