己酸二乙氨基乙醇酯的致突变性研究

采用鼠伤寒沙门氏菌回复突变试验(Ames试验),用TA97,TA98,TA100和TA102菌株,加与不加S9,剂量分别设为每皿1000μg,500μg和250μg;小鼠骨髓多染红细胞微核试验,剂量设为雄性、雌性小鼠给药剂量均为250,500和1000mg*kg-1;小鼠睾丸精母细胞染色体畸变试验,剂量设为625mg*kg-1,1250mg*kg-1和2500mg*kg-1.结果表明,Ames试验中,各测试浓度的诱发回变菌落数均未超过自发回变菌落数的2倍;小鼠骨髓多染红细胞微核试验和小鼠睾丸精母细胞染色体畸变试验,各剂量组和溶剂对照组的微核率进行统计学处理,未见有显着性差异(P＞0.05),表明各项试验结果均为阴性.因此,己酸二乙氨基乙醇酯无致突变作用.     

饮用水遗传毒性测试中TA98和TA100的敏感度和可靠度比较

为了寻找适合我国水质特征的Ames试验简化方法,作者通过广泛的文献调研,对近30年以来,我国研究者用Ames试验评价饮用水和水源水的遗传毒性的实验数据进行了总结和分析。结果发现,TA98-S9测试体系的敏感度和可靠度最高,TA98+S9次之,TA100-S9与TA100+S9 2个测试体系对Ames试验结果的影响几乎可以忽略。只用1个TA98-S9测试体系的方法 1,仅用25%的工作量,可获得91%的致突变阳性检出率和74%的有效数据率;采用TA98-S9和TA98+S9 2个测试体系的方法 2,使用50%的工作量,可获得96%的阳性检出率和96%的有效数据率。据此,作者建议:采用Ames试验评价我国饮用水和水源水的遗传毒性时,可以将通常采用的4个测试体系的方法 4,简化为只采用TA98-S9与TA98+S9 2个测试体系的方法 2。在仅进行致突变性定性评价时,或者在费用或水样量不足等条件下,则可使用更为简化的方法 1,更利于Ames试验的广泛应用。     

官厅水库沉积物极性提取液化学成分分析及基因毒性效应

提取北京市官厅水库表层沉积物极性组分,定量检测了12种硝基芳香烃和5种芳香胺的含量水平;并通过鼠伤寒沙门氏菌回复突变试验(Ames试验)研究沉积物极性提取液的致突变毒性,对官厅水库沉积物的潜在基因毒性效应进行评价.结果表明,官厅水库沉积物中12种硝基芳香烃的总含量为8.4—272 ng·g~(-1),平均值为121 ng·g~(-1);5种芳香胺总含量为5.1—42.8 ng·g~(-1),平均值为4.8 ng·g~(-1).各组沉积物极性提取液对鼠伤寒沙门氏菌(Salmonella typhimurium)TA98及TA100的致突变毒性试验结果均呈阴性,但多个点位沉积物极性提取液导致的细菌突变数随提取液浓度的上升而增加,表明可能存在致突变物质.Pearson相关分析结果表明,2-硝基甲苯、3-硝基甲苯、1,3,5-三硝基苯和联苯胺的含量与TA98的突变效应具有显著相关性,推断其可能是主要的致突变物质,且其可能引起的致突变类型为移码型基因突变.     

2-硝基芴的致突变性受共存多环芳烃的影响

本文以Ames试验的TA_(98)菌株研究了2-硝基芴(2-Nitroflurene)在其它多环芳烃共存时,其致突变性的变化.结果显示:它与其它硝基多环芳烃共存时.直接致突变性增强;而与其它无直接致突变性的多环芳烃共存时,其直接致突变性减弱,且多环芳烃的环数越多、减弱作用越强,这可能是因为硝基多环芳烃具有亲电子性,DNA的碱基有亲核性,易于结合,造成DNA的损伤;而多环芳烃一般具有供电子性,阻碍了硝基多环芳烃与DNA碱基的结合.     

Ames测试不确定性分析

Ames测试具有实验周期短、简便的优点,是目前普遍采用的检测化合物致突变性的初筛方法,但Ames测试仍有一定的不确定性.本文主要依据本实验室的研究结果分析了Ames测试中测试菌株、统计回复子数目的时间及2倍判定标准给测试结果带来的不确定性,对影响Ames测试结论存在假阳性及假阴性的因素做了进一步分析与讨论.图6表1参19     

工业碳黑的致突变性研究

本文研究了工业碳黑有机溶剂萃取物的致突变性.发现其萃取物在Ames试验中显示极强的直接致突变性,特别是经过硝化工艺的碳黑.而且萃取溶剂不同,其致突变性也不同,以氯苯和二氯甲烷为溶剂的萃取物活性较高.经进一步分析测定,在萃取物中含有二硝基芘和3-硝基芴酮等硝基多环芳烃,前者是一种非常强的直接致突变物,它们是工业碳黑致突变性的主要供献者,光照可以降低碳黑萃取物的致突变活性.     

荧光假单胞菌(Pseudomonas fluorescens)工程菌株田间残留和扩散的追踪检测

分别在北京和江苏省连云港市对携带Btcry基因的荧光假单胞菌工程菌进行了田间残留与扩散的追踪检测 .对越冬前后的试验地和保护地土壤样品进行抗生素抗性平板分离 ,能检测到极少量的具有与出发菌株相同抗性的荧光假单胞菌菌落 ,但没有发现工程菌株的残留与扩散 .对江苏试验地样品还进行了工程菌株质粒卡那霉素和壮观霉素抗性标记基因的抗性菌落分离 ,绝大多数样品中都能分离到包括荧光假单胞菌在内的抗性菌落 ,土样中菌密度n(cfu) =10 4 ～ 10 5g-1,但进行Btcry基因PCR -RFLP检测时没有从样品中得到特异扩增产物 .研究结果表明 :工程菌株抗性标记基因在自然界广泛存在 ,工程菌在株环境中没有残留和扩散 ,具有良好的生物安全性 .表 4参 8     

Tet抗性铜绿假单胞菌接合转移pHN102方法研究

二亲本杂交实验通常要求受体菌不能携带有与质粒上相同的抗性.本次实验中受体菌铜绿假单胞菌(Pseudo-monas aeruginosa)和重组质粒pHN102都带有四环素(Tc)抗性.考虑到质粒pHN102导入受体菌后至少带有两个Tc抗性基因片段,转移接合子的抗性会增强,所以提高四环素浓度至20μg/mL,并利用pHN102上携带的luxAB发光酶基因标记受体菌筛选转移接合子,并设立对照菌株.通过抽提质粒、琼脂糖凝胶电泳检测,确定pHN102成功转入P.aeruginosa中.转移接合子经4次转接后,质粒仍可以稳定存在且能够稳定表达.P.aeruginosa(pHN102)的发光动力学曲线表明,加入底物20min后,发光强度趋于稳定.经液体培养稀释后进行发光度和活菌数测定,发现二者呈显著的线性关系(r=0.994).图4表2参6     

高盐度工业废水优势降解菌的生长特性及应用

从3处不同的污泥中筛选到4株对高盐度工业废水有较好降解效果的菌株A41、A45、TA2、XA3。通过生长特性研究确定出4株菌株的最佳温度、pH、盐度适应范围,A41分别为(35℃,7和2%~3%),A45分别为(35℃,6和0.5%~4%),TA2分别为(35℃,8和0.5%~4%),XA3分别为(45℃,7和6%~8%)。试验结果表明,当A41、TA2、XA3的接种量按1∶1∶1混合时,处理工业废水效果最好。3株混合菌株处理高盐度废水的最佳处理时间为24 h、COD去除率达到了50.5%;最佳pH为9,COD去除率达到了58.3%。     

基于luxAB标记菌株的发光定量分析及其在镉毒性检测方面的应用

以E.coli(pHN102)为研究对象,探讨了luxAB标记菌株的发光定量方法以及影响其发光测定的相关因素.结果表明,该标记菌株在癸醛诱导发光后存在动态变化的发光曲线,发光强度随时间变化先升后降.增大癸醛浓度会提高发光强度,当癸醛浓度达到1%时出现最大发光值;供试菌株在35℃和pH6条件下的发光强度最大,与E.coli的最适生长条件相一致.应用此标记菌株检测镉(Cd)毒性,其相对发光度与Cd浓度存在负相关,线性回归分析可达到1%显著水平.Cd在LB培养基、M9培养基和水中的EC50值分别为4.27mmol/L、34.8mmol/L和62μmol/L,说明Cd在水介质中的毒性远大于在LB、M9培养基中的毒性.图6表2参16     

Antimutagenic and bacteriostatic activities of Schiff‐base compounds derived from aminoguanidine,semicarbazone and thiosemicarbazone and a copper(II)‐coordination complex

The Schiff‐base aminoguanidine compounds including a resorcylidene aminoguanidine copper(II) complex that were synthesized at our laboratory posses non‐mutagenic properties when tested with Ames test using Salmonella typhimurium TA97, TA100 and TA102 bacterial strains. These compounds were tested as possible pharmacological agents in preventing a wide array of illnesses. Additionally, some of these compounds exhibit strong antimutagenic and bacteriostatic activity. Nitrovin, a known mutagen, was used in the antimutagenicity tests as the inducer. A three‐dimensional projection of the copper(II) complex was derived by the semi‐empirical method ZINDO/1 to obtain additional information about its structure, and to help elucidate a possible mechanism of action.     

Evaluation of the antimutagenic and antiradical potentials of extracts from the tubers of (Tunisian) Cyperus rotundus

The mutagenic potential of aqueous, total oligomers flavonoids (TOF), ethyl acetate and methanol extracts from tubers of Cyperus rotundus L. was assessed using Ames Salmonella tester strains TA98 and TA100, and SOS chromotest strain Escherichia coli PQ37 with and without metabolic activation (S9). None of the different extracts produced a mutagenic effect. Likewise, the antimutagenicity of the same extracts was tested using the “Ames test” and the “SOS chromotest”. Our results showed that C. rotundus extracts possess antimutagenic effects against S. typhimurium TA98 and TA100 strains towards the mutagen aflatoxin B1 (AFB1), similar to E. coli PQ37 strain against AFB1 and Nifuroxazide mutagens using the SOS chromotest assay. A free radical scavenging test was used in order to explore the antioxidant capacity of the extracts obtained from the tubers of C. rotundus. TOF, ethyl acetate and methanol extracts showed an important free radical scavenging activity towards the 1,1-diphenyl-2-picrylhydrazine (DPPH) free radical. These extracts showed an IC50 value of respectively 5, 20 and 65?µg?mL^?1. The beneficial effects of TOF, ethyl acetate and methanol extracts of C. rotundus were assessed by antioxidant and antimutagenic activities.     

Synthesis and biological activities of new azacrown ether Schiff bases and spectrophotometric studies of their complexation with [60]fullerene

A series of novel azacrown ether Schiff bases 1–3 have been synthesized in good yield and in a simple way. Their host–guest interaction with [60]fullerene has been studied in toluene by absorption spectroscopic method. All the complexes are found to be stable with 1:1 stoichiometry. Because of their potential applications in industry, agriculture and medicine, they were investigated for their mutagenic and antimutagenic activities using the spot test and the plate incorporation assay of Ames. Compounds 1, 2 and 3 were found to be nonmutagenic in the Ames test using strains TA 1535, TA100 and TA97a of Salmonella typhimurium. However, using strain TA102 revealed that, although both compounds 1 and 2 were nonmutagenic, compound 2 gave a positive response indicating that it acts as an oxidative mutagen. The structure-activity relationship may throw some light on the biological activity of such series of compounds.     

Aliphatic and aromatic halocarbons as potential mutagens in drinking water

Chlorinated phenols are either products of industrial chemical processes or the result of chlorination of drinking water. Often, the formation of chlorinated phenols is based upon naturally occurring phenol. The following chlorinated phenols have been selected for testing in the Ames‐test for their mutagenic activity: 3‐chlorophenol, 4‐chlorophenol, 2,3,6‐, 2,4,5‐, 2,4,6‐trichlorophenol, 4‐chloro‐2‐methylphenol and 4‐chloro‐3‐methylchlorophenol. The tester strains TA97, TA98, TA100 and TA104 were employed. All tested compounds produced mutagenic activity at least in one tester strain. The highest numbers of revertants were detected for 2,3,6‐ and 2,4,6‐trichlorophenol. But in contrast to the other substances, these two induced only frameshift mutations in presence of a metabolizing system. The evidence of their presence in drinking water and of their mutagenic activity makes them to a potential health hazard.     

Mutagenicity of diesel engine exhaust in the Ames / Salmonella assay using a direct exposure method

The aim of this study was to investigate the potential mutagenic activity of diesel engine exhaust in the Ames/Salmonella assay using a direct aerosol exposure system. So, TA 98 and TA 100 strains, with or without added S9 mix, were exposed to diesel emissions after varying degrees of filtration. Variants of these two strains, deficient in nitroreductase (TA 98NR and TA 100NR) or over-expressing O-Acetyl Transferase (YG 1024 and YG 1029), were also exposed to total (unfiltered) diesel exhaust to highlight the putative mutagenicity of any nitro-PAHs present in these emissions. Mutagenic activity of the diesel exhaust was demonstrated on Salmonella typhimurium, strains TA 100 and variants TA 100 NR and YG1029. The use of a particle filter did not modify the genotoxicity of the diesel emissions, indicating a major contribution of the gas phase to the mutagenicity of these diesel emissions. The prominent role of the particulate-associated nitro-polycyclic aromatic hydrocarbons (nitro-PAHs) claimed by some authors working on diesel exhaust organic extracts was not confirmed by our results with native diesel exhaust exposure. Our results show that the gas phase is potentially more mutagenic than the particles alone.     

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.     

Evaluation of mutagenicity associated with Escherichia coli inactivation in UV-treated orange juice

UV radiation is an alternative technology for the elimination of pathogenic micro-organisms in liquid food. The objectives of this work were to investigate mutagenesis using the Ames test in : (1) fresh squeezed orange juice submitted to UV radiation doses required to achieve 5 and 10 log₁₀ reductions of different strains of Escherichia coli serotype O157:H7 at two temperatures, and (2) commercial pasteurized orange juice submitted to thermal treatment.

Two histidine-auxotrophic strains, Salmonella Typhimurium TA98 and TA97a, were used in the Ames test with and without metabolic activation.

In commercial pasteurized orange juices, mutagenesis was not observed with S. Typhimurium TA98 strain with and without metabolic activation, but high mutagenic effect was observed using TA97a with metabolic activation.

UV radiation did not affect the stability of ascorbic acid in aqueous solution. However, degradation of ascorbic acid was observed when UV treatment was performed on squeezed orange juice, following first order kinetics.

UV treatment required to achieve 5 log₁₀ reductions (1.5 J/cm²) did not show a mutagenic effect with and without metabolic activation using the tested strains, but a higher UV radiation dose necessary to produce 10 log₁₀ reductions (3.0 J/cm²) showed mutagenicity with metabolic activation.     

Modification of mutagenic activity of benzo[a]pyrene by trace elements in the Ames assay

Benzo[a]pyrene (BaP) is one of the most ubiquitous environmental contaminants and exerts potent carcinogenicity and mutagenicity. In the presence of rat liver S9 mix, four trace elements, germanium (Ge), iron (Fe), zinc (Zn), and molybdenium (Mo), were included in the Ames Salmonella microsome/mutagenic assay to test their inhibitory effects on BaP-induced mutations. All trace elements were found to exert an inhibitory effect in tester TA98 and TA100 strains; Fe was the most potent among the elements tested, whereas Zn was weakly inhibitory to prevent mutations. The present findings demonstrate that the antimutagenic potential of trace elements, notably Fe, may be attributed to its modulatory effect on the bioactivation and detoxification processes of BaP.     

Silver nanoparticle antimicrobial activity explained by membrane rupture and reactive oxygen generation

Silver nanoparticles are widely used as antimicrobial compounds based on empirical observations. However, there is few knowledge on the mechanism ruling the antimicrobial activity and toxicity of Ag nanoparticles. Here, we investigated this mechanism. Nano-Ag was synthesised by thermal co-reduction. Mutagenicity analysis was performed using Salmonella typhimurium histidine auxotrophic strains TA 98 and TA 100 at nano-silver concentrations of 100 to 500 µg per plate. Dose-dependent analysis for reactive oxygen species generation has been performed using 2,7-dichlorofluorescein diacetate dye. Membrane integrity has been analyzed at 260 nm, before and after treatment. We also used scanning electron microscopy, membrane permeabilization test, and superoxide formation determinations. Results show that the average particle size of Ag nanoparticle is 60.4 ± 3.8 nm. The minimum inhibitory concentration of Ag nanoparticles for E.coli is 30 µg/mL; the minimum bactericidal concentration is 40 µg/mL. Ames mutagenicity tests showed negative results, which may be explained by the antimicrobial activity of nano-silver. Bacterial inner wall were indeed ruptured, and cytoplasmic content was released after 5 min of treatment in a dose-dependent manner. We thus propose that reactive oxygen generation and alteration of membrane integrity and permeability are the major mechanism of antimicrobial activity of nano-silver.