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.     

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。     

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.     

Nitro-Deficient Niclosamide Confers Reduced Genotoxicity and Retains Mitochondrial Uncoupling Activity for Cancer Therapy

Niclosamide is an oral anthelmintic drug, approved for use against tapeworm infections. Recent studies suggest however that niclosamide may have broader clinical applications in cancers, spurring increased interest in the functions and mechanisms of niclosamide. Previously, we reported that niclosamide targets a metabolic vulnerability in p53-deficient tumours, providing a basis for patient stratification and personalised treatment strategies. In the present study, we functionally characterised the contribution of the aniline 4′-NO₂ group on niclosamide to its cellular activities. We demonstrated that niclosamide induces genome-wide DNA damage that is mechanistically uncoupled from its antitumour effects mediated through mitochondrial uncoupling. Elimination of the nitro group in ND-Nic analogue significantly reduced γH2AX signals and DNA breaks while preserving its antitumour mechanism mediated through a calcium signalling pathway and arachidonic acid metabolism. Lipidomics profiling further revealed that ND-Nic-treated cells retained a metabolite profile characteristic of niclosamide-treated cells. Notably, quantitative scoring of drug sensitivity suggests that elimination of its nitro group enhanced the target selectivity of niclosamide against p53 deficiency. Importantly, the results also raise concern that niclosamide may impose a pleiotropic genotoxic effect, which limits its clinical efficacy and warrants further investigation into alternative drug analogues that may ameliorate any potential unwanted side effects.     

电子烟基因毒性评价方法概述

为了解电子烟的基因毒性及其评价方法,按照受试对象分类进行了综述。其中以细菌为受试对象的试验方法有细菌回复突变试验和DNA损伤分析等;以离体细胞为受试对象的试验方法有微核试验、DNA双链断裂试验、RNA转录测序试验、定向基因检测分析和Bhas细胞转化试验等;以模式动物为受试对象的试验方法有长期吸入毒性试验和生殖发育毒性试验等;以人体为受试对象的主要是临床试验和流行病学调查研究等。由于受试对象和试验方法的差异性,导致电子烟基因毒性结果的可信度和可比性较差,因此建议在评价电子烟基因毒性的过程中,应至少考虑细菌、离体细胞、模式动物和临床试验等4个层次的受试对象,通过回复突变试验、微核试验、长期吸入毒性试验和临床试验等多种试验方法获得多个基因毒性的试验终点,科学、客观和全面地综合评价电子烟的基因毒性。     

昆仑雪菊遗传毒性研究

目的研究昆仑雪菊的遗传毒性,为其食用安全性提供科学依据。方法试验受试物为昆仑雪菊浸泡浓缩液,每毫升相当于1 g昆仑雪菊干品。Ames试验和体外哺乳类细胞染色体畸变试验分别设昆仑雪菊浸泡浓缩液0.008、0.04、0.2、1、5 mg/皿组和1.25、2.5、5 mg/ml组,直接计数培养基上各菌株的回变菌落数和染色体畸变细胞数。微核试验、精子畸形试验和小鼠精原细胞或精母细胞染色体畸变试验均设昆仑雪菊浸泡浓缩液低、中、高剂量组(5、10、20 g/kg),各试验分别镜检计数每只动物1 000个嗜多染红细胞(PCE)、1 000个完整精子、100个中期分裂相细胞,记录有微核的嗜多染红细胞数量、畸变精子类型和数目、染色体畸变细胞数。结果昆仑雪菊遗传毒性试验结果均为阴性。结论本研究未发现昆仑雪菊有遗传毒性。     

紫色杆菌素及脱氧紫色杆菌素的基因毒性评价

目的：考察紫色杆菌素、脱氧紫色杆菌素以及二者混合物的基因毒性,为其更广泛的活性研究和应用奠定基础。方法：利用SOS/umu检测试剂盒定量分析紫色杆菌素、脱氧紫色杆菌素、二者混合物以及各自的S9代谢产物是否刺激鼠伤寒沙门菌Salmonella typhimurium NM2009细胞产生了SOS修复,以及与阳性药物2-氨基芴（2-aminofluorene,AF-2）和2-氨基蒽（2-aminoanthracene,2-AA）进行对比所产生毒性作用的强弱。结果：32.6 μg/mL的脱氧紫色杆菌素、经S9代谢后的34.2 μg/mL的紫色杆菌素和33.4 μg/mL的混合物对S. typhimurium NM2009细胞产生了基因毒性,但与阳性对照物相比属于低毒性物质,且相同剂量的紫色杆菌素和脱氧紫色杆菌素以及二者混合物的基因毒性基本相同,二者混合后未产生协同效应。结论：紫色杆菌素和脱氧紫色杆菌素属于低基因毒性物质。     

Relevance of protein fermentation to gut health

It is generally accepted that carbohydrate fermentation results in beneficial effects for the host because of the generation of short chain fatty acids, whereas protein fermentation is considered detrimental for the host's health. Protein fermentation mainly occurs in the distal colon, when carbohydrates get depleted and results in the production of potentially toxic metabolites such as ammonia, amines, phenols and sulfides. However, the effectivity of these metabolites has been established mainly in in vitro studies. In addition, some important bowel diseases such as colorectal cancer (CRC) and ulcerative colitis appear most often in the distal colon, which is the primary site of protein fermentation. Finally, epidemiological studies revealed that diets rich in meat are associated with the prevalence of CRC, as is the case in Western society. Importantly, meat intake not only increases fermentation of proteins but also induces increased intake of fat, heme and heterocyclic amines, which may also play a role in the development of CRC. Despite these indications, the relationship between gut health and protein fermentation has not been thoroughly investigated. In this review, the existing evidence about the potential toxicity of protein fermentation from in vitro animal and human studies will be summarized.     

Relative Genotoxicities of PAH of Molecular Weight 252 AMU in Coal Tar-Contaminated Sediment

Bioassay-directed chemical fractionation methodology was used to calculate relative mutagenic potencies of polycyclic aromatic hydrocarbons (PAH) of molecular weight 252 amu in coal tarcontaminated sediment from Sydney Harbour, Nova Scotia. A normal phase HPLC technique was used to separate organic solvent extracts into fractions containing isomeric PAH of a single benzologue class. Bioassays with Salmonella typhimurium strain YG1025 with the addition of oxidative metabolism (S9) showed that approximately 50% of the mutagenic activity observed in the sediment extract was associated with PAH of molecular weight 252 amu. Further separation of the 252 PAH fraction using reversed phase HPLC yielded subfractions containing individual compounds; bioassay dose-response curves for these subfractions showed that benzo[a]pyrene was responsible for approximately 75% of the activity of the 252 PAH fraction.