Complex Mechanisms of Antimony Genotoxicity in Budding Yeast Involves Replication and Topoisomerase I-Associated DNA Lesions,Telomere Dysfunction and Inhibition of DNA Repair

Antimony is a toxic metalloid with poorly understood mechanisms of toxicity and uncertain carcinogenic properties. By using a combination of genetic, biochemical and DNA damage assays, we investigated the genotoxic potential of trivalent antimony in the model organism Saccharomyces cerevisiae. We found that low doses of Sb(III) generate various forms of DNA damage including replication and topoisomerase I-dependent DNA lesions as well as oxidative stress and replication-independent DNA breaks accompanied by activation of DNA damage checkpoints and formation of recombination repair centers. At higher concentrations of Sb(III), moderately increased oxidative DNA damage is also observed. Consistently, base excision, DNA damage tolerance and homologous recombination repair pathways contribute to Sb(III) tolerance. In addition, we provided evidence suggesting that Sb(III) causes telomere dysfunction. Finally, we showed that Sb(III) negatively effects repair of double-strand DNA breaks and distorts actin and microtubule cytoskeleton. In sum, our results indicate that Sb(III) exhibits a significant genotoxic activity in budding yeast.     

Abnormal chromosome assessment of snakehead fish (Channa striata) affected by heavy metals from a reservoir near an industrial factory

This study aimed to investigate nine heavy metal concentrations (As, Cd, Cr, Pb, Cu, Fe, Zn, Mn and Ni) in water, sediment and snakehead fish (Channa striata) and to identify abnormal chromosomes in C. striata from a reservoir near an industrial factory and a reference area. Heavy metal concentrations were measured by inductively coupled plasma optical emission spectrometry (ICP-OES). The metal concentrations in the water, sediment and C. striata samples did not exceed the standard limit of Thailand, except for Cr concentrations, which exceeded water quality standards. The concentrations of Cd, Fe, Mn and Ni in C. striata samples between the reservoir and the reference area were significantly different (p < 0.05). The diploid chromosome number of C. striata from both areas was (2n = 42). Eight types of abnormal chromosomes were identified and classified as a single-chromatid gap, a single-chromatid break, centric fragmentation, a centric gap (CG), fragmentation, deletion, single-chromatid decomposition and iso-arm fragmentation. The most frequent abnormal chromosome in the samples was CG. The percentages of abnormal chromosomes in the C. striata samples from the reservoir near the industrial factory and the reference area were significantly different (p < 0.05) at 8.44 and 1.20, respectively.     

Evaluation of chlorite and chlorate genotoxicity using plant bioassays and in vitro DNA damage tests

In the last few years chlorine dioxide has been increasingly used for disinfecting drinking water in many countries. Although it does not react with humic substances, chlorine dioxide added to water is reduced primarily to chlorite and chlorate ions, compounds that are under investigation for their potential adverse effects on human health. The aim of this research was to study the genotoxicity of chlorite and chlorate and their mixtures. The end-points included two plant tests (chromosomal aberration test in Allium cepa and micronucleus assay in Tradescantia, carried out at different times of exposure) and two genotoxicity tests in human HepG2 cells (comet assay and cytokinesis-blocked micronucleus test). Preliminary toxicity tests were carried out for both plant and HepG2 assays. The results showed that chlorite and chlorate are able to induce chromosomal damage to plant systems, particularly chromosomal aberrations in A. cepa root tip cells, even at concentrations lower than the limit established by Italian normative law and WHO guidelines. In HepG2 cells increased DNA damage was only observed for chlorate at the lowest concentration. No increase in micronuclei frequency was detected in any of the samples tested in human HepG2 cells.     

Genotoxicity in water and sediment extracts from the St Lawrence river system, using the SOS chromotest

Surface water and sediments from the St Lawrence River system (Québec region) were analysed for genotoxicity using the SOS Chromotest, as well as for their chemical concentrations of polycyclic aromatic hydrocarbons and heavy metals. Additionally, chlorobenzenes, polychlorinated biphenyls, organochlorinated pesticides, ammonia and nitrite concentrations in sediments were determined. Water and sediments sampled from twenty-five sites were initially partitioned into their aqueous and particulate phases by tangential flow filtration and centrifugation, respectively. Organic contaminants were extracted from the fractions with dichloromethane. For surface water, fifteen extracts of filtered water and seven of particulates, and for sediments one extract of pore water and three of particulates proved to be weakly genotoxic. All but one of the genotoxic responses observed in the surface water were obtained from samples taken from the highly industrial portion of the St Lawrence River system, with the strongest responses observed in Lake St Louis. Surface water genotoxicants partitioning favours the particulate fraction. Bottom particulates genotoxicity was 1000-fold weaker than suspended particulates, on a per unit weight basis. Additionally, whole sediments were extracted with a 10% dimethylsulfoxide-saline solution. The observed distributions of genotoxicity values did not correlate with observed concentrations of demonstrated SOS inducers, mutagens and/or carcinogens, nor with the presence of other toxic chemicals.     

贯筋藤凝乳剂对小鼠急性毒性与遗传毒性研究

目的探究贯筋藤作为凝乳剂的安全性。方法以贯筋藤凝乳剂为受试物,采用限量法评价贯筋藤凝乳剂对小鼠的急性毒性,观察小鼠体重及解剖后心脏、脾脏、肝脏、肾脏是否发生病变;采用Ames实验评价其遗传毒性,设20、100、500、2500、12500μg/皿剂量组进行遗传毒性实验。结果急性毒性实验未出现小鼠死亡情况。研究发现小鼠的精神亢奋,有较强攻击欲望,根据小鼠的生理状况对小鼠进行脑组织中MAO-A和5-HT含量检测发现空白组和实验组差异极显著(P0.01)。通过Ames试验检测贯筋藤凝乳剂的遗传毒性,根据结果发现在12500μg/皿的情况下出现抑制微生物生长的情况,而在2500μg/皿的剂量下统计每组菌落数差异不著性(P0.05)。结论贯筋藤凝乳剂的LD50剂量大于10000 mg/kg,并根据Ames试验确定贯筋藤凝乳剂在最高2500μg/皿不具有致突变作用。     

Hemocompatibility,cytotoxicity, and genotoxicity of poly(methylmethacrylate)/nanohydroxyapatite nanocomposites synthesized by melt blending method

The aim of this study was to show the hemocompatibility, cytotoxicity, and genotoxicity of nanocomposites that were synthesized with different molecular weights of poly(methyl methacrylate) (PMMA) and different concentrations of nanohydroxyapatite (nHAp). Different techniques to characterize the nanocomposites were used. The cytotoxicity and genotoxic effects of the polymers and nanocomposites on human lymphocytes were determined by acid phosphatase assay, viability test, and comet assay. Moreover, hemocompatibility test was performed. It was found that all of the PMMA/nHAp nanocomposites are highly hemocompatible and biocompatible, none of the nanocomposites showed a cytotoxic effect, and nHAp addition decreased the genotoxicity.     

Lab‐on‐a‐Chip‐Based High‐Throughput Screening of the Genotoxicity of Engineered Nanomaterials

The continuous increasing of engineered nanomaterials (ENMs) in our environment, their combinatorial diversity, and the associated genotoxic risks, highlight the urgent need to better define the possible toxicological effects of ENMs. In this context, we present a new high‐throughput screening (HTS) platform based on the cytokinesis‐block micronucleus (CBMN) assay, lab‐on‐chip cell sorting, and automated image analysis. This HTS platform has been successfully applied to the evaluation of the cytotoxic and genotoxic effects of silver nanoparticles (AgNPs) and silica nanoparticles (SiO₂NPs). In particular, our results demonstrate the high cyto‐ and genotoxicity induced by AgNPs and the biocompatibility of SiO₂NPs, in primary human lymphocytes. Moreover, our data reveal that the toxic effects are also dependent on size, surface coating, and surface charge. Most importantly, our HTS platform shows that AgNP‐induced genotoxicity is lymphocyte sub‐type dependent and is particularly pronounced in CD2+ and CD4+ cells.     

Chlorinated Polycyclic Aromatic Hydrocarbons Associated with Drinking Water Disinfection: Synthesis,Formation under Aqueous Chlorination Conditions and Genotoxic Effects

Polycyclic aromatic hydrocarbons (PAHs) are among the most persistent and toxic organic micropollutants present in water and several of them are mutagenic and carcinogenic. Although it has been shown that chlorinated derivatives of PAHs (Cl-PAHs) may be formed during the water chlorination procedure, little is known about their potential genotoxic and carcinogenic effects. The objectives of the present work were to prepare and characterize the major chlorinated derivatives of benzo[a]pyrene (BaP) and fluoranthene (Fluo), to develop an analytical methodology for their quantification in water samples and to analyse their potential genotoxicity. Chlorinated standards were prepared by a newly developed two phase method (water/n-hexane) using sodium hypochlorite. 6-Chloro-benzo[a]pyrene was selectively obtained from BaP, while 1,3-dichloro-fluoranthene and 3-chloro-fluoranthene were obtained from Fluo. All products were isolated and characterized by nuclear magnetic resonance and mass spectrometry. The formation of BaP- and Fluo-chlorinated derivatives under aqueous chlorination conditions was observed using a SPE-HPLC-FLD methodology. In addition, the cytotoxic and genotoxic activities of the three chlorinated derivatives were analyzed in comparison to their parent compounds, in a human-derived hepatoma cell line using the neutral red uptake and comet assays, respectively. The results showed that, at the equimolar doses of 100 and 125 μM, 6-Cl-BaP was able to induce a significantly higher level of DNA damage than BaP, suggesting a more potent genotoxic effect. In contrast, neither Fluo nor its chlorinated derivatives were genotoxic in the same cell line. The identification of new and possibly hazardous water chlorination by-product from PAHs emphasizes the need to minimize total organic carbon content of raw water and the implementation of safer water disinfection methods.