Anticlastogenic action of vitamin C against genotoxicity of estrogenic drug diethylstilbestrol (DES) in the human lymphocyte chromosomes

Several sex steroids and estrogenic drugs are genotoxic in varying conditions and cause oxidative stress, which has been a field of interest to study the molecular mechanism of the genetic damage. Among the estrogenic drugs, a strong toxic effect is exerted by diethylstilbestrol (DES). In the present study it has been attempted to study its genotoxic effects in human lymphocyte assay system along with ameliorative or anticlastogenic effects of vitamin C. The drug was used with different dosage of concentrations on human lymphocytes administered in vitro. The parameters used were Sister Chromatid Exchanges (SCEs) and Chromosomal Aberrations (CAs). Higher levels of clastogeny and SCEs have been observed indicating significant damaging effect by the drug. Interesting ameliorating effects were observed in the presence of vitamin C which is a well-known antioxidant. The results support the possibility of practical application of natural protectors against the mutagenic/oenotoxic action of chemical mutagens.     

Protective role of nordihydroguaiaretic acid (NDGA) against the genotoxic damage induced by ethynodiol diacetate in human lymphocytes in vitro

Antioxidants and plant products are reported to reduce the genotoxic damage of steroids. In our present study we have tested different dosages of nordihydroguaiaretic acid (NDGA) against the genotoxic damage induced by ethynodiol diacetate in the presence of S9 mix. Treatments with nordihydroguaiaretic acid (NDGA) results in the reduction of the genotoxic damage. A significant decrease was observed at all the tested doses of NDGA in sister chromatic exchanges of number of abnormal cells. The results suggest a protective role of NDGA against the genotoxic damage.     

常乐稀土复合肥对玉米（Zea may L.）的细胞毒性和遗传毒性效应

为了研究常乐稀土多元复合肥对主要农作物--玉米(Zea mays L.)是否产生细胞毒性和遗传毒性效应,采用不同梯度质量浓度的稀土复合肥溶液对玉米进行处理,观察其根尖生长状况,统计分析细胞死亡率、微核率、有丝分裂指数以及对细胞DNA的损伤情况.结果表明,质量浓度为10mg·L-1以下时,常乐稀土复合肥能够促进玉米根尖的生长,100mg·L-1以上时抑制根尖生长;随着试验浓度的递增,细胞死亡率、微核率、DNA损伤率逐渐上升并具有正相关性,表现出明显的剂量-效应关系,而有丝分裂指数却随着浓度的增加逐渐下降.以上结果表明,常乐稀土复合肥对玉米确有一定的细胞毒性和遗传毒性效应,其细胞毒性阈值和遗传毒性阈值皆为100mg·L-1.     

Genotoxicity study of phenol and o-cresol using the micronucleus test and the comet assay

Phenol and cresols are common toxic environmental pollutants. In this study, the micronucleus assay and the alkaline single cell gel electrophoresis (SCGE) technique have been used to investigate the genotoxic activity of phenol and o-cresol. The result of experiments revealed that phenol and o-cresol were both evident genotoxins, and the genotoxic activity of o-cresol was stronger. In the micronucleus assay, phenol and o-cresol could both cause a significant increase in the micronucleus (MN) frequencies (p?p?p?p?相似文献   

Genotoxic effects of two different classes of insecticide in developing chick embryos

Pesticides provide considerable protection against pest population; however, rampant accumulation of these chemicals into varied habitats across the globe necessitates the need for a careful screening of each chemical due to toxic manifestations. In the current study, the genotoxic potential of two different classes of commercial insecticides – chlorpyrifos and cypermethrin combination and Spinosad, a naturalyte were compared. Rhode Island Red chick embryos were exposed to different doses of either of these insecticides individually, by in ovo treatment. Genotoxicity was then evaluated through micronucleus (MN) test and Comet assay. The combination insecticide exposure at low doses of 0.05 and 0.1 μg/egg induced DNA damage as evidenced by an increased tail moment in the Comet assay. Further, the presence of micronucleated erythrocytes and also various abnormal cells including dacryocytes, microcytes, erythroplastids, squashed/notched nuclei, and spindle-shaped erythrocytes in the blood smear consolidates indicate the presence of insecticide-induced genotoxicity. Spinosad, however, was found only mildly genotoxic but at a high dose of 1.5 mg/egg. The results indicate that usage of naturalyte insecticide may be a better option to minimize the harmful effects of chemical insecticides.     

Role of GSH in the amelioration of chromium-induced membrane damage

Membrane damage is one of the important consequences of chromium (Cr), an environmental toxicant, induced cytotoxicity. Reduced glutathione (GSH), a membrane protectant may be used to reduce the Cr-induced membrane damage. In the present study, the impact of Cr in presence and absence of GSH was studied on plasma membrane of the liver and kidneys in male Wistar rats. Significant increases in membrane cholesterol levels as well as significant decreases in membrane phospholipid levels in Cr exposed (0.8 mg per 100 g body weight, i.p., for 28 days) animals suggest structural alterations in both the liver and kidney plasma membranes. Alkaline phosphatase (ALP), total ATPase, and Na⁺–K⁺–ATPase activities of plasma membrane were significantly decreased in both the liver and kidneys after Cr treatment. This treatment also produced significant weight loss and increased Cr content in the liver and kidneys. However, GSH (8 mg per 100g body weight, i.p., daily at an interval of 6 h after injection of Cr for a period of 28 days) supplementation restored alterations induced by Cr in plasma membrane of both the liver and kidneys but was not able to eliminate the deposited Cr from the liver and kidney tissues.     

人血淋巴细胞检测浊漳河地表水的遗传毒性

为评价浊漳河水体中有机污染物的致突变性,采用单细胞凝胶电泳(SCGE)技术,研究了浊漳河水体3个不同样点的有机提取物对人外周血淋巴细胞DNA损伤效应。结果显示,各水样中的有机提取物均能引起DNA损伤,且随剂量的增高,有机物对DNA的损伤加重;细胞DNA损伤与对照组相比均有显著性差异(p<0.01)。这表明人外周血淋巴细胞对水体有机污染物的毒作用非常敏感。细胞彗星尾部DNA含量(%DNAT)和染毒剂量呈极显著正相关(R2>0.8547,p<0.01)。以上结果表明,浊漳河水体受到一定程度的有机物污染,这些有机污染物可诱导淋巴细胞DNA损伤,具有遗传毒性。SCGE技术作为一种简便、快速和灵敏的检测方法在水环境遗传毒性监测方面具有较大的应用价值。     

The microcystins-induced DNA damage in the liver and the heart of zebrafish,Danio rerio

Microcystins (MCYST) are the freshwater cyanobacterial toxins, known to induce hepatocellular carcinoma, necrosis, intrahepatic bleeding, as well as human and livestock mortality. Within hepatocytes, MCYST selectively bind to protein phosphatases 1 and 2A, resulting in severe liver damage. The toxicology of MCYST in mice and rats has been well studied, but little is known regarding genotoxicity in aquatic animals. In this study, the zebrafish, Danio rerio was exposed to crude extract of Microcystis aeruginosa bloom. Liver and heart were examined for MCYST-induced toxicity. Light microscopy at 36?h revealed severe, widespread apoptotic necrosis of the majority of hepatocytes, and cytoskeletal deformation in myocardiocytes. Hepatocytes were dissociated with cell shrinkage and margination of nuclear chromatin. Laddering of genomic DNA from the liver and heart of the exposed fish in an increment of 180–200?bp was consistent with apoptosis. Fluorimetric analysis of DNA unwinding was carried out to determine the DNA strand breakage. After 36?h exposure, the % double-stranded DNA was significantly reduced in hepatocytes and myocardiocytes. In conclusion, the results obtained in this study indicate that, the extract of M. aeruginosa bloom is genotoxic to fish. The DNA damage observed in this study may be attributed to the activation of DNA endonucleases. This model of DNA damage may contribute for identifying novel molecular mechanisms of interest for therapeutic application.     

An anticlastogenic in vivo micronucleus assay for tea

Common use of antimutagens and anticarcinogens in everyday life is an effective measure for preventing human cancer and genetic diseases. Antioxidant properties of tea have vast potential as protective agents against diverse toxic effects. The present study was aimed to evaluate the role of aqueous clonal tea extracts (green tea, oolong tea and black tea) in modulating the genotoxic damage induced by cyclophosphamide (CP), a commonly used chemotherapeutic drug and a well-known mutagen and clastogen. All the three tea extracts at 1 and 2% concentration did not increase the frequency of micronucleated polychromatic erythrocytes (MPE) in bone marrow cells of mice when administered individually. The tea extracts decreased the micronuclei (MN) induced by CP. Therefore, regular intake of tea may improve the antioxidant status in in vivo and thereby reduce the risk of cancer and coronary heart disease.     

In situ evaluation of the genotoxic potential of the river Nile: I. Micronucleus and nuclear lesion tests of erythrocytes of Oreochromis niloticus niloticus (Linnaeus, 1758) and Clarias gariepinus (Burchell, 1822)

This study aimed to investigate the genotoxic potential of chemicals present along the course of the river Nile using frequencies of micronuclei (MN) and nuclear lesions (NL) in erythrocytes of Nile tilapia Oreochromis niloticus niloticus and African catfish Clarias gariepinus, as biomarkers. Results showed that most of the physicochemical parameters detected and heavy metal concentrations were significantly higher in the water collected from the estuaries of the river Nile compared to other sites of the upper Nile. The frequencies of MN and NL in peripheral blood erythrocytes of Nile tilapia and African catfish were significantly higher in estuary sites in Damietta and Rosetta compared to upper sites. The lowest level of genotoxicity was observed at two sites (Aswan and Kena), considered to be less contaminated. Our results suggested that higher frequencies of MN and NL determined at Damietta and Rosetta sites may be indicative of damage produced by pollutants in these areas. The most remarkable result was that MN and NL frequencies appear to be strongly related to water quality at different sites examined, indicating that MN frequencies may serve as a reliable biomarker for testing genotoxicity in situ. The positive correlation between MN and NL induction suggested that NL may be a useful complementary assay for genotoxicity analyses when fish are used as experimental animals. It was also found that seasonal variations in MN and NL frequencies might contribute to a better understanding of genotoxic responses in the field. The use of fish as indicator organisms for monitoring the presence of genotoxic-inducing contaminants in the environment seemed justified because the effects of exposure to a “complex mixture” such as river water were obtained. Nile tilapia appears to be a more suitable bioindicator species than African catfish in studying genotoxic chemical pollution in the river Nile attributed to a higher sensitivity.     

玉米种子的DNA指纹计算机化鉴定

对１２个优良玉米自效系的ＤＮＡ进行了ＲＡＰＤ分析，共筛选了２５０个Ｏｐｅｒｏｎ引物，其中１２个引物共扩增出５９个多态性产物，根据这５９个多态性ＲＡＰＤ产物，对１２个自交系进行了聚类分析，把它们分为３个群，结果与根据系谱法的聚类结果一致。经过优选，用ＯＰＮ－１１扩增出的１１种ＤＮＡ带，建立了这１２个自交系的ＲＡＰＤ－ＤＮＡ指纹图谱。在该图谱呼弱个自交系都具有特异的ＤＮＡ指纹，可以与其它自效纱相区别。     

过敏症状儿童室内PM_(2.5)对小鼠巨噬细胞的氧化损伤及VE的抗氧化保护作用研究

越来越多的研究提示,主要空气污染物PM_(2.5)暴露浓度的升高与儿童过敏性疾病的发病率有着密切的关系,然而PM_(2.5)暴露与过敏性疾病之间的关联尚未完全阐明。为探究患有过敏症状儿童的室内PM_(2.5)对小鼠巨噬细胞的氧化损伤作用以及维生素E(vitamin E,VE)的抗氧化保护作用,从5户患有1种或1种以上的过敏性症状(如过敏性鼻炎、哮喘)儿童的室内采集PM_(2.5),分别考察了不同剂量PM_(2.5)暴露24 h后如何影响小鼠巨噬细胞的氧化应激水平,指标包括活性氧(ROS),还原型谷胱甘肽(GSH),丙二醛(MDA),8-羟基脱氧鸟苷(8-OH-dG),以及炎症因子水平,指标包括肿瘤坏死因子ɑ(TNF-ɑ),白介素8β(IL-8β)的影响。结果表明,200μg·mL~(-1)PM_(2.5)暴露组与对照组比较,细胞内ROS积累,出现脂质过氧化以及DNA损伤,并伴有炎症反应的发生,差异均有统计学意义(P0.05或P0.01);VE(50 mg·mL~(-1))+200μg·mL~(-1)PM_(2.5)组的ROS、MDA、8-OHdG、TNF-ɑ、IL-8β含量低于200μg·mL~(-1)PM_(2.5)组,GSH含量高于200μg·mL~(-1)PM_(2.5)组。较高剂量(200μg·mL~(-1))PM_(2.5)可诱导小鼠腹腔巨噬细胞出现氧化损伤,VE在该应激过程中起着一定的保护作用。     

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.     

PFOA对斑马鱼胚胎发育、行为和DNA损伤的毒性研究

全氟辛酸(PFOA)是一种分布广泛的环境持久性有机污染物,在野生动物与人体内普遍被检出。目前PFOA发育毒性的特点和机制尚未阐明,已有的研究显示,PFOA毒性与受试物种及性别密切相关。因此,利用不同物种进行PFOA毒性研究对阐明其毒性机制十分重要。本研究选用了斑马鱼这种理想的脊椎动物模型,考察了PFOA暴露对其胚胎发育的自主运动、心跳、行为、细胞凋亡和DNA损伤的影响。研究发现,始于6hpf的PFOA暴露会导致斑马鱼胚胎自主运动异常,心率降低,且具有一定的剂量依赖性;6～24hpf的高浓度的PFOA暴露(>414.0mg·L-1)会导致胚胎发生显著的细胞凋亡,凋亡主要出现在眼部、头部、心脏和尾部。较高浓度的PFOA暴露(165.6mg·L-1)会使仔鱼的光刺激应激行为模式发生变化。此外,PFOA暴露会致使斑马鱼发生DNA损伤,且损伤程度随PFOA浓度的升高而加重,表明PFOA具有一定的基因毒性。上述结果表明,PFOA对斑马鱼胚胎具有发育毒性,具体表现为自主运动异常,心率降低和行为反应能力变弱,同时伴随畸形、细胞凋亡和DNA损伤。     

应用超敏感酵母HUG1-yEGFP生物传感器筛选基因毒性化合物

为了扩展本实验室构建的响应于DNA损伤信号的超敏感酵母HUG1-yEGFP生物传感器的检测范围并积累参考数据,为实际应用打下实验基础,本研究对26种与环境及人类生活息息相关的化合物的基因毒性进行了检测,筛选出8种具有基因毒性的化合物,并将其结果与现有微生物检测系统的检测结果进行了对比。结果显示,超敏感酵母HUG1-yEGFP生物传感器的检测结果与以细菌为宿主细胞的Ames试验或SOS显色法的检测结果的吻合度高于50%,与以酵母为宿主细胞的GSA检测系统的检测结果的吻合度高于85%。以真核生物酵母为宿主的检测系统与以原核生物细胞为宿主的检测系统的结果的差异反映了它们抗胁迫机制的不同。本研究应用超敏感酵母HUG1-yEGFP生物传感器特异性检测出3种化合物为基因毒性阳性,这3种化合物是代森锰锌、孔雀石绿及丙烯酰胺,均被报道具有潜在致癌性。本研究结果表明,超敏感酵母HUG1-yEGFP生物传感器可以有效地对环境污染有关的基因毒性化合物进行检测,可以作为其他现有检测系统的补充,应用于环境化学污染的健康风险评价。     

Genotoxic potential of organic extracts from municipal reclaimed water in Tianjin,China

Despite increasing pressure to make more efficient use of water resources, the widespread use of reclaimed water still remains a contentious issue, primarily because of risks to human health arising from water pollution by organic compounds. Therefore, safety evaluation of reclaimed water is an urgent need. The aim of this study was to determine the genotoxic potential of reclaimed water during low and high water periods. Reverse phase C-18 solid-phase extraction (RP-C18SPE) was used for the extraction of organic compounds from water samples. The tests, namely, Ames, micronucleus and chromosome aberrations were used to determine the damage caused by water samples on genetic material. Ames tests showed that both influent and effluent reclaimed water, except influent after metabolism by S9 in high water period, induced mutation of TA98 and TA100 strains of Salmonella typhimurium in a concentration-response manner. There were no significant differences in micronucleus and chromosome aberration tests. Even after treatment, reclaimed water in Tianjin still showed mutational effects and new strategies for reduction of genotoxins need to be considered.     

Indium tin oxide nanoparticles-mediated DNA fragmentation and cell death by apoptosis in human lung epithelial cells

Indium tin oxide (ITO) nanoparticles (NP) have extensive applications in industrial fields, and concerns regarding their potential toxicity in humans and environmental impact have increased. Since exposure to ITO NP is mainly via skin and inhalation, this study was conducted utilizing human lung epithelial (A549) cell line. Cells were exposed to different concentrations of the ITO NP for 24 and 48 hr. A severe cytotoxic response of ITO NP was observed as evident by the (3-4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and neutral red uptake assays after 48 hr exposure. ITO NP significantly reduced glutathione levels with a concomitant increase in lipid hydroperoxide levels, superoxide activity, and reactive oxygen species (ROS) generation after exposure. A significant induction in caspase activity and formation of condensed chromosomal bodies was also observed after ITO NP (10 or 25 µg/ml) exposure. Furthermore, a significant induction in DNA damage was observed by the Comet assay in cells exposed to ITO NP. Our data demonstrate that ITO NP display cytotoxic and genotoxic potential. However, increase in ROS levels and oxidative stress leading to oxidative DNA damage and condensed chromosomal bodies formation, suggests involvement of apotosis. Thus, ITO NP-mediated effects on cell viability indicate cytotoxicity, and therefore, exposures need to be carefully monitored in the industrial sector.     

Genotoxicity and mutagenicity of electronic waste leachates using animal bioassays

Human exposure to electronic waste (e-waste) is increasing, especially in developing nations. This has potential short and chronic adverse health risks. Information is currently scarce on potential genotoxicity of e-waste leachate. In this study, the potential mutagenic and genotoxic effects of leachates from e-waste dumpsites at two electronic markets in Lagos, Nigeria, were investigated. The assays utilized were: bone marrow micronucleus (MN) and chromosome aberration (CA), spermatogonia CA, sperm morphology, and sperm count in mice. Experiments were carried out at concentrations of 1%, 5%, 10%, 25%, and 50% (v/v; leachate: distilled water) of leachate samples. MN analysis showed a concentration-dependent induction of micronucleated polychromatic erythrocytes (MNPCE) across the treatment groups. In the CA tests (bone marrow and spermatogonia), there was concentration-dependent significant reduction in mitotic index and induction of different types of CA. Assessment of sperm shape showed a significant increase in sperm abnormalities with significant decrease in mean sperm count in treated groups. Heavy metals analyzed in the tested samples are believed to contribute significantly to the observed genetic damage. This indicates that e-waste contains potential genotoxic agents; and constitutes a genetic risk in exposed human population.     

Supramolecular complex formation,a possible antigenotoxic mechanism of glucomannan against aflatoxin B1

Abstract

Glucomannan is a highly branched polysaccharide with glycosidic linkages, constituted of mannoses and glucoses. In recent years, its usefulness due to its immunological, antioxidant and antimutagenic activity has been recognized. The aim of the study was to determine the antigenotoxic ability of glucomannan extracted from Candida utilis orally administered (100–700?mg/kg) to mice, which subsequently received 1?mg/kg aflatoxin B₁. Hepatocytes obtained from these animals 4–16?h post administration were examined by means of the comet assay. The antigenotoxic effect was found to be higher than that observed in previous studies with α-mannan and β-D-glucan isolated from Saccharomyces cerevisiae., In order to explore the possibility of formation of a supramolecular complex between glucomannan and aflatoxin B₁, both compounds were co-crystallized, their melting points determined, and the complex analyzed through ultraviolet spectroscopy. The spectroscopy data suggest that the protective effect of glucomannan is related to the formation of a supramolecular complex between the two compounds.     

Nanosilica exerts cytotoxicity and apoptotic response via oxidative stress in mouse embryonic fibroblasts

Nanoscale silica is an important industrial material and extensively used in medicines. The objective of this study was to determine potential cytotoxicity and genotoxic effects attributed to nanosilica exposure in mouse embryonic fibroblasts (L929) cells. Nanosilica produced mild cytotoxicity in L929 cells. Results showed that nanosilica increased thiobarbituric acid reactive substance levels and enhanced superoxide dismutase activity but decreased levels of glutathione. This was accompanied by a concomitant generation of reactive oxygen species, loss of mitochondrial membrane potential, and activation of caspase-3 activity. In addition, in the single-cell gel test, nanosilica (50–300 μg/ml) at two treatment times 24 and 48 hr produced concentration- and time-dependent increase of DNA damage. Therefore, the obtained results indicate that nanosilica may induce genotoxic effects in cultured L929 cells associated with induction of oxidative stress.