Mutagenicity In TA98 Research Articles

Analysis of mutagenic components of oxidative hair dyes with the Ames test

Oxidative hair dyes consist of two components (I and II) that are mixed before use. Aromatic amines in component I and their reaction with hydrogen peroxide after mixing them with component II have been of primary concern. In addition, two in vitro genotoxicity assays are still required for the approval of the final products of oxidative hair dyes in China, and the substance in the oxidative hair dye that causes the high rate of positive results in genotoxicity tests, especially the Ames test, has not been fully elucidated. In this study, we analyzed the formulation of 55 different oxidative hair dyes from 7 color series and performed Ames tests in the strain TA98 with the S9 mix (oxidative hair dyes No. 1-30) and in strain TA97a without the S9 mix (oxidative hair dyes No. 31-55). We found that toluene-2,5-diamine sulfate (2,5-diaminotoluene sulfate, DATS) in component I may be the cause of mutagenicity in TA98, and hydrogen peroxide in component II may be the cause of mutagenicity in TA97a, and their positive concentrations were consistent with those that we calculated from Ames tests. The results suggest that the positive results for the oxidative hair dye in the Ames test were inevitable because of the existence of DATS in component I and of hydrogen peroxide in component II. Therefore, we should carry out safety assessments on each raw material and carry out risk assessments on the final products of oxidative hair dyes instead of genotoxicity tests in China.

Screening of rhizomes of Rheum emodi Wall. Ex. Meissen for antimutagenic potential employing Ames assay

Rheum emodi Wall. Ex. Meissen is an important food and medicinal plant growing wild in western Himalayas. The current endeavor was undertaken to carry out bioassay guided fractionation and isolation of antimutagenic principle(s) present in the rhizomes of Rheum emodi. The antimutagenicity assay was performed to check the recuperating effect of 80% aqueous methanol extract of rhizomes of R. emodi and its fractions i.e. n-hexane, chloroform, ethyl acetate, n-butanol and water fractions, against 4-nitro-o-phenylenediamine (NPD), sodium azide, and 2AF-induced mutagenicity in TA98 and TA100 strains using the Ames Salmonella his+ reversion assay. The crude 80% aqueous methanol extract and ethyl acetate fraction exhibited strong antimutagenicity. The antimutagenic potential of the extracts and fractions was markedly enhanced when indirect acting mutagen was used in both the tester strains of Salmonella typhimurium. In an effort to identify the molecules responsible for the antimutagenic effect displayed by R. emodi, four anthraquinones i.e. aloe-emodin, chrysophanol, emodin and rhein detected in the ethyl acetate fraction by HPLC analysis were also tested for their antimutagenic potential in both the tester strains. It was found that the anthraquinones in their pure form showed lower antimutagenicity than the mother fraction from which they were isolated. The results point towards a synergistic effect of the principles present in the mother fraction.

In-vitro antioxidant, antimutagenic and cancer cell growth inhibition activities of Rhododendron arboreum leaves and flowers.

In the current investigation, the active principles of the methanol extracts of Rhododendron arboreum leaves (MEL) and flowers (MEF) were investigated with the help of ultra-high performance liquid chromatography (UHPLC), amino acid analyzer and gas chromatography mass spectrometry (GC-MS). UHPLC revealed different polyphenols present in the extracts. GC-MS identified 20 phytochemicals in leaves and 17 in the flowers, whereas, amino acid analyzer confirmed 11 amino acids in leaves and 10 in the flowers. The extracts were subjected to the investigation of biological activity through analysis of antioxidant activity in different in vitro assays, antimutagenic activity in Ames assay and cancer cell growth inhibition activity by MTT (3-4,5 dimethylthiazol-2,5 diphenyltetrazolium bromide) assay. MEL showed higher antioxidant activity in lipid peroxidation inhibition assay (95.32 ± 0.37%) than MEF (77.09 ± 4.17%) with IC50103.6 µg/ml for MEL and 271.17 µg/ml for MEF. In nitric oxide scavenging assay, an activity of 94.46 ± 0.32% (IC50 150.13) was observed in MEF followed by 83.71 ± 0.74% (IC50 179.52) in MEL. The antimutagenic activity of both the extracts was evaluated against sodium azide, 4-nitro-O-phenylenediamine and 2-aminofluorene mutagens in TA-98 and TA-100 strains of Salmonella typhimurium. The analysis was carried out using pre- and co-incubation modes. However, both extracts were observed to possess considerable antimutagenic activity against different known mutagens, flowers came out to be more effective than the leaves in terms of % inhibition. The extracts also exhibited significant cancer cell growth inhibition activity, when tested against 3 cancer cell lines namely, Human cervical cancer cell line (HeLa), Breast cancer cell line (MCF7) and Lung cancer cell line (A549). In case of HeLa and A549, MEL showed higher activity of 64.62 ± 2.65 and 75.08 ± 1.68% as compared to 53.11 ± 2.84 and 45.92 ± 2.43% in MEL, respectively. The EC50 values for MEL in HeLa and A549 were noted to be 232.76 and 155.38 µg/ml, respectively, whereas, MEF had EC50 of 395.50 µg/ml in HeLa and 660.26 µg/ml in A549. Further, MEF showed higher cytotoxicity in MCF7 cell line (84.93 ± 1.17%) followed by the MEL (73.57 ± 1.27%) with EC50 value of 95.16 µg/ml for MEF followed by 172.19 µg/ml for MEL. The biological activities of the extracts can be attributed to the phyto-constituents identified by sophisticated instruments.The biological activities of the extracts can be attributed to the active principles identified by sophisticated instruments.

Identification of methylglyoxal as a major mutagen in wood and bamboo pyroligneous acids.

To identify the major mutagen in pyroligneous acid (PA), 10 wood and 10 bamboo pyroligneous acids were examined using the Ames test in Salmonella typhimurium strains TA100 and TA98. Subsequently, the mutagenic dicarbonyl compounds (DCs), glyoxal, methylglyoxal (MG), and diacetyl in PA were quantified using high-performance liquid chromatography, and the mutagenic contribution ratios for each DC were calculated relative to the mutagenicity of PA. Eighteen samples were positive for mutagens and showed the strongest mutagenicity in TA100 in the absence of S9 mix. MG had the highest mutagenic contribution ratio, and its presence was strongly correlated with the specific mutagenicity of PA. These data indicate that MG is the major mutagen in PA.

Cytotoxicity and mutagenicity assessment of organomodified clays potentially used in food packaging

Modern food packaging has made great advances as result of global trends and consumer preferences, which are oriented to obtain improved food quality and safety. In this regard, clay minerals, and mainly Montmorillonite (Mt) are attracting considerable interest in food packaging because of the improvements developed in mechanical and barrier properties. Hence, the present work aim to assess the toxicity of four Montmorillonite-based clay minerals, an unmodified clay, Cloisite®Na+ (CNa+), and three modified Mt clays: Cloisite®30B (C30B), a commercial clay, and Clay1 and Clay2, two novel modified organoclays developed by the Packaging, Transport, & Logistics Research Institute (ITENE). First, the cytotoxic effects were studied in the Human Umbilical Vein Endothelial Cells (HUVEC). In addition, the potential mutagenicity of the clays was evaluated by the Ames test. Clay1 did not induce any cytotoxic effects in HUVEC, although it exhibited potential mutagenicity in TA98 Salmonella typhimurium strain. In contrast, Clay2 produced cytotoxicity in endothelial cells but no mutagenicity was recorded. However, CNa+ was not cytotoxic neither mutagenic. And finally, C30B showed positive results in both assays. Therefore, results showed that clay minerals have a different toxicity profile and a case by case toxicity evaluation is required.

Comparative mutagenicity and genotoxicity of particles and aerosols emitted by the combustion of standard vs. rapeseed methyl ester supplemented bio-diesel fuels: Impact of after treatment devices: Oxidation catalyst and particulate filter

Diesel exhausts are partly responsible for the deleterious effects on human health associated with urban pollution, including cardiovascular diseases, asthma, COPD, and possibly lung cancer. Particulate fraction has been incriminated and thus largely investigated for its genotoxic properties, based on exposure conditions that are, however, not relevant for human risk assessment. In this paper, original and more realistic protocols were used to investigate the hazards induced by exhausts emitted by the combustion of standard (DF0) vs. bio-diesel fuels (DF7 and DF30) and to assess the impact of exhaust treatment devices (DOC and DPF). Mutagenicity and genotoxicity were evaluated for (1) resuspended particles (“off line” exposure that takes into account the bioavailability of adsorbed chemicals) and for (2) the whole aerosols (particles+gas phase components) under continuous flow exposure (“on line” exposure). Native particles displayed mutagenic properties associated with nitroaromatic profiles (YG1041), whereas PAHs did not seem to be involved. After DOC treatment, the mutagenicity of particles was fully abolished. In contrast, the level of particle deposition was low under continuous flow exposure, and the observed mutagenicity in TA98 and TA102 was thus attributable to the gas phase. A bactericidal effect was also observed in TA102 after DOC treatment, and a weak but significant mutagenicity persisted after DPF treatment for bio-diesel fuels. No formation of bulky DNA-adducts was observed on A549 cells exposed to diesel exhaust, even in very drastic conditions (organic extracts corresponding to 500μg equivalent particule/mL, 48h exposure). Taken together, these data indicate that the exhausts issued from the bio-diesel fuels supplemented with rapseed methyl ester (RME), and generated by current diesel engines equipped with after treatment devices are less mutagenic than older ones. The residual mutagenicity is linked to the gas phase and could be due to pro-oxydants, mainly for RME-supplemented fuels.

Mutagenicity of the cysteine S-conjugate sulfoxides of trichloroethylene and tetrachloroethylene in the Ames test

The nephrotoxicity and nephrocarcinogenicity of trichloroethylene (TCE) and tetrachloroethylene (PCE) are believed to be mediated primarily through the cysteine S-conjugate β-lyase-dependent bioactivation of the corresponding cysteine S-conjugate metabolites S-(1,2-dichlorovinyl)-l-cysteine (DCVC) and S-(1,2,2-trichlorovinyl)-l-cysteine (TCVC), respectively. DCVC and TCVC have previously been demonstrated to be mutagenic by the Ames Salmonella mutagenicity assay, and reduction in mutagenicity was observed upon treatment with the β-lyase inhibitor aminooxyacetic acid (AOAA). Because DCVC and TCVC can also be bioactivated through sulfoxidation to yield the potent nephrotoxicants S-(1,2-dichlorovinyl)-l-cysteine sulfoxide (DCVCS) and S-(1,2,2-trichlorovinyl)-l-cysteine sulfoxide (TCVCS), respectively, the mutagenic potential of these two sulfoxides was investigated using the Ames Salmonella typhimurium TA100 mutagenicity assay. The results show both DCVCS and TCVCS were mutagenic, and TCVCS exhibited 3-fold higher mutagenicity than DCVCS. However, DCVCS and TCVCS mutagenic activity was approximately 700-fold and 30-fold lower than DCVC and TCVC, respectively. DCVC and DCVCS appeared to induce toxicity in TA100, as evidenced by increased microcolony formation and decreased mutant frequency above threshold concentrations. TCVC and TCVCS were not toxic in TA100. The toxic effects of DCVC limited the sensitivity of TA100 to DCVC mutagenic effects and rendered it difficult to investigate the effects of AOAA on DCVC mutagenic activity. Collectively, these results suggest that DCVCS and TCVCS exerted a definite but weak mutagenicity in the TA100 strain. Therefore, despite their potent nephrotoxicity, DCVCS and TCVCS are not likely to play a major role in DCVC or TCVC mutagenicity in this strain.

The effect of a novel tobacco process on the in vitro cytotoxicity and genotoxicity of cigarette smoke particulate matter

Some of the toxic effects of smoking have been attributed to the combustion of nitrogenous protein in tobacco. The effects of a treatment which reduces tobacco’s protein nitrogen level, on the in vitro cytotoxicity and genotoxicity of cigarette smoke particulate matter (PM), were measured. PMs were tested in the Neutral Red Uptake (NRU) test; the Salmonella mutagenicity assay (SAL); the mouse lymphoma mammalian cell mutation assay (MLA) and the in vitro micronucleus test (IVMNT). PMs from all of the cigarettes were cytotoxic and genotoxic. PM obtained from smoking treated tobacco, showed a small, consistent and statistically significant reduced mutagenicity (revertants/μg) in TA98 with post-mitochondrial supernatant (S9). No consistent quantitative or qualitative differences were detected in the other tests. The data are discussed in relation to published information on smoke chemistry obtained from cigarettes made of tobacco treated using this technique. The observations confirm that the method did not give rise to any new qualitative or quantitative cytotoxic or genotoxic effects, and may have reduced PM’s bacterial mutagenicity in TA98 with S9. Further toxicity testing is warranted, to investigate the effects of the tobacco treatment in more detail and add to the data already obtained.

In vitro genotoxicity of neutral red after photo-activation and metabolic activation in the Ames test, the micronucleus test and the comet assay

Neutral red (Nr) is relatively non-toxic and is widely used as indicator dye in many biological test systems. It absorbs visible light and is known to act as a photosensitizer, involving the generation of reactive oxygen species (type-I reaction) and singlet oxygen (type-II reaction).The mutagenicity of Nr was determined in the Ames test (with Salmonella typhimurium strains TA1535, TA97, TA98, TA98NR, TA100, and TA102) with and without metabolic activation, and with and without photo-activation on agar plates. Similarly to the situation following metabolic activation, photo-mutagenicity of Nr was seen with all Salmonella strains tested, albeit with different effects between these strains. To our knowledge, Nr is the only photo-mutagen showing such a broad action. Since the effects are also observed in strains not known to be responsive to ROS, this indicates that ROS production is not the sole mode of action that leads to photo-genotoxicity. The reactive species produced by irradiation are short-lived as pre-irradiation of an Nr solution did not produce mutagenic effects when added to the bacteria. In addition, mutagenicity in TA98 following irradiation was stronger than in the nitroreductase-deficient strain TA98NR, indicating that nitro derivatives that are transformed by bacterial nitroreductase to hydroxylamines appear to play a role in the photo-mutagenicity of Nr. Photo-genotoxicity of Nr was further investigated in the comet assay and micronucleus test in L5178Y cells. Concentration-dependent increases in primary DNA damage and in the frequency of micronuclei were observed after irradiation.

The Antimutagenic Effect of Mistletoe Lectin (Viscum album L. var. coloratum agglutinin)

A galactose- and N-acetyl-D-galactosamine-specific lectin (Viscum album L. var. coloratum agglutinin, VCA), which is known for its anticancer activity, was isolated from mistletoe. In this study, we investigated the antimutagenic potentials of VCA by using the pre-incubation method of the Ames test (Salmonella typhimurium TA98 and TA100) in the presence or absence of S9 mixture. Viscum album L. var. coloratum agglutinin was assessed for its antimutagenic properties against the mutagens 2-aminoanthracene (2AA) and furylfuramide (AF-2) for strain TA98, and sodium azide (NaN(3) ) and 2-aminoanthracene (2AA) for strain TA100. The concentrations used for this test compound were 100, 200 and 400 µg per plate. Viscum album L. var. coloratum agglutinin showed moderate, but not negligible, protective effects regarding the antimutagenic properties against the direct-acting mutagens NaN(3) and AF-2. Furthermore, VCA was more effective in preventing the mutagenicity of the indirect-acting mutagen 2-AA (in the presence of S9) when tested with both TA98 and TA100. In conclusion, this report has shown broad ranging antimutagenic effects of VCA to numerous mutagens in TA98 and TA100 Salmonella typhimurium strains. Although the data presented here cannot be applied in vivo, they can support other antimutagenic and anticarcinogenic findings for VCA.

1-Methoxy-3-indolylmethyl glucosinolate; a potent genotoxicant in bacterial and mammalian cells: Mechanisms of bioactivation

1-Methoxy-3-indolylmethyl (1-MIM) glucosinolate, contained in many Brassica vegetables, is strongly mutagenic in Salmonella typhimurium TA100 when activated by myrosinase. Here, we describe the synthesis and evaluation of two breakdown products, 1-MIM nitrile and 1-MIM alcohol. 1-MIM nitrile was not mutagenic and 1-MIM alcohol showed low direct mutagenicity in TA100, indicating that other breakdown products mediated the mutagenicity of 1-MIM glucosinoate/myrosinase in this strain. However, 1-MIM alcohol was strongly mutagenic to a TA100-derived strain expressing human sulphotransferase SULT1A1. Likewise, 1-MIM glucosinolate (with myrosinase) showed 10 times higher mutagenic activity in TA100-SULT1A1 than in strain TA100. Identical adducts, N 2-(1-MIM)-dG and N 6-(1-MIM)-dA, were detected in both strains, but the levels were higher in TA100-hSULT1A1. A similar influence of SULT1A1 was observed in recombinant V79-hSULT1A1 cells compared to parental SULT-deficient Chinese hamster V79 cells. 1-MIM glucosinolate (with myrosinase) as well as 1-MIM alcohol induced sister chromatid exchange in both cell lines, but with clearly higher efficiency in V79-hSULT1A1 cells. Gene mutation assays were conducted at the HPRT locus with 1-MIM alcohol in V79-hSULT1A1 cells, and with 1-MIM glucosinolate/myrosinase in V79 parental cells. In both cases, the result was clearly positive. Thus, 1-MIM glucosinolate is mutagenic in bacterial and mammalian cells via at least two different metabolites.

Synthesis and characterization of selected 4,4′-diaminoalkoxyazobenzenes

The role of the –N(CH 2CH 2OH) 2 group in producing a mutagenic response from 4-((3-(2-hydroxyethoxy)4-amino)phenylazo)- N, N-bis(2-hydroxyethyl)aniline has been investigated. To accomplish this goal, a group of substituted 4,4′-diaminoazobenzene dyes was synthesized, and their structures were confirmed using 1H NMR, TOF-LC-ESI mass spectrometry, and combustion analysis. Mutagenicity was determined using the standard Ames test in Salmonella strains TA98, TA100, and TA1538 with and without S9 enzyme activation. The results of this study provide evidence that the mutagenicity of the parent dye arises from the metabolic cleavage of N-hydroxyethyl groups to give the corresponding –NHCH 2CH 2OH and –NH 2 substituted monoazo dyes as direct-acting mutagens. All 5 of the dyes studied were mutagenic at various levels with and without S9 enzyme activation in TA1538. In addition, the results show that removing one N-hydroxyethyl group and capping both –OH groups in the parent dye did not affect mutagenicity, whereas removing both N-hydroxyethyl groups produced a strong direct-acting mutagen in all three bacterial strains. Increasing the length of the N-alkyl chain from two to three carbon atoms eliminated mutagenicity in TA98 without S9 activation.

Chemical fate and mutagenic formation potentials of phenothiazine and related compounds during water chlorination

The reactions of hetero-tricyclic aromatic hydrocarbons (H-TCAHs) with hypochlorite in an aqueous solution were investigated under conditions that simulate wastewater disinfection. H-TCAH-hypochlorite reaction products were determined by gas chromatographic-mass spectrometric (GC-MS) analyses. For 20 µM, 10H-phenothiazine, 10H-phenoxazine, and phenoxathiin reacted rapidly with active chlorine in neutral pH (7.0), but no phenazine-hypochlorite reaction was observed over pH values of 5-9 for 1 hr. The 10H-phenothiazine-hypochlorite reaction began by oxidation with active chlorine to form its dioxides, followed by chloro-substitution in water. The extent of the reactions depended on the chlorine dose, solution pH and compound structures. Ames assays for the chlorination byproducts of 10H-phenothiazine and 10H-phenoxazine also showed to be weak mutagenicity in TA98 and TA100 strains without S9 mix, but no chlorination byproducts of phenoxathiin exhibited any mutagenicity in both tester strains with and without S9 mix.

Mutagenicity of surface soil from residential areas in Kyoto city, Japan, and identification of major mutagens

To clarify the mutagenic potential of surface soil in residential areas in Kyoto city, surface soil samples were collected twice or three times from 12 sites, and their organic extracts were examined by the Ames/ Salmonella assay. Almost all (>92%) samples showed mutagenicity in TA98 without and with S9 mix, and 8/25 (32%) samples showed high (1000–10,000 revertants/g of soil) or extreme (>10,000 revertants/g of soil) activity. Moreover, to identify the major mutagens in surface soil in Kyoto, a soil sample was collected at a site where soil contamination with mutagens was severe and continual. The soil extract, which showed potent mutagenicity in TA98 without S9 mix, was fractionated by diverse column chromatography methods. Five major mutagenic constituents were isolated and identified to be 1,6-dinitropyrene (DNP), 1,8-DNP, 1,3,6-trinitropyrene (TNP), 3,9-dinitrofluoranthene (DNF), and 3,6-dinitrobenzo[ e]pyrene (DNBeP) by co-chromatography using high performance liquid chromatography and spectral analysis. Contribution ratios of 1,6-DNP, 1,8-DNP, 1,3,6-TNP, 3,9-DNF, and 3,6-DNBeP to total mutagenicity of the soil extract in TA98 without S9 mix were 3, 10, 10, 10, and 6%, respectively. These nitroarenes were detected in surface soil samples collected from four different residential sites in other prefectures, and their contribution ratios to soil mutagenicity were from 0.7 to 22%. These results suggest that surface soil in residential areas in Kyoto was widely contaminated with mutagens and there were some sites where surface soils were heavily polluted. 1,6-DNP, 1,8-DNP, 1,3,6-TNP, 3,9-DNF, and 3,6-DNBeP may be major mutagenic constituents that contaminate surface soil in Kyoto and other residential areas.

Biodirected mutagenic chemical assay of PM 10 extractable organic matter in Southwest Mexico City

The concentration of breathable particles (PM 10) in urban areas has been associated with increases in morbidity and mortality of the exposed populations, therein the importance of this study. Organic compounds adsorbed to PM 10 are related to the increased risk to human health. Although some studies have shown the lack of correlation between specific mutagenic compounds in an organic complex mixture (OCM) and the mutagenic response in several bioassays, the same organic compounds selectively separated in less complex groups can show higher or lower mutagenic responses than in the OCM. In this study, we fractionated the OCM, from the PM 10 in four organic fractions of increasing polarity (F1–F4). The Salmonella bioassay with plate incorporation was applied for each one using TA98, with and without S9 (mammalian metabolic activation), and YG1021 (without S9) strains. The most polar fraction (F4) contained the greatest mass followed by F1 (non-polar), F2 and F3 (moderately polar). The concentrations of the OCM as well as the F4 were the only variables correlated with PM 10, atmospheric thermal inversions, fire-prone area, NO 2, SO 2, CO, rain and relative humidity. This indicated that polar organic compounds were originated in gas precursors formed during the atmospheric thermal inversions as well as the product of the incomplete combustion of vehicular exhausts and of burned vegetation. The percentages of the total PAH, and the individual PAH with molecular weight ≥ 228 g mol −1 (except retene) correlated with the percentages of indirect-acting mutagenicity in TA98 + S9. The percentages of the total nitro-PAH and most of the analyzed individual nitro-PAH correlated with percentages of the direct-acting mutagenicity in both TA98 − S9 and YG1021, the latter being more sensitive. In general, the highest mutagenic activity (indirect and direct) was found in F3 (moderately polar) and in F4 (polar). The non-polar fraction (F1) did not exhibit any kind of mutagenicity. In 77% of the cases, mutagenic activity was higher in the sum fractions with respect to their OCM. The combinations between F1, F2 and F4, with F3 under different or equal proportions suggested that mutagenicity reduction, in the combined matter of January (with TA98 + S9 and YG1021) and of May (with YG1021), was due to concentrations of mutagens and non-mutagens in each fraction, and not to an antimutagenic effect. The organic compounds present in the non-polar fractions showed no antagonism, inhibition or reduction in the most mutagenic fractions in both indirect- and direct-acting mutagenicity, and the less polar organic compounds in F3 reduced mutagenicity in F4, in both months.

Evaluation of extracellular products and mutagenicity in cyanobacteria cultures separated from a eutrophic reservoir

The algal extracellular products (ECPs) in three cultures of cyanobacteria species (Anabaena, Microcystis, and Oscillatoria) dominating the eutrophic reservoir populations and their toxins have been investigated in the present work. Using gas chromatography coupled with high-resolution electron-impact mass spectrometry (GC/EI-MS) and high performance anion-exchange chromatography (HPAEC) techniques, more than 20 compounds were found in the algal culture (including cells and filtrates) extracts. The main identified ECPs were classified to polysaccharides, hydrocarbons, and aldehydes. Odor causing substances such as trans-1,10-dimethyl-trans-9-decalol (geosmin) and 2-methylisoborneol (2-MIB)were also found in the algal cultures. The potential mutagenicity of the algal suspensions was also studied with the Ames test. The organic extracts of the algal suspension from the axenic cultures were mutagenicity in TA98 without S9 mix and in TA100 with and without S9 mix. The results indicate that the ECPs of three algae species dominating the eutrophic reservoir were mutagenic clearly in the bacterial test.

The role of steric effects in the direct mutagenicity of N-acyloxy- N-alkoxyamides

Electrophilic N-acyloxy- N-alkoxyamides are mutagenic in Salmonella typhimurium TA100 without the need for S9 metabolic activation and they react with DNA at guanine—N7 at physiological pH. Since these are direct-acting mutagens, structural factors influence binding and reactivity with DNA. Mutagenicity in TA100 can be predicted by a QSAR incorporating hydrophobicity (log P), stability to substitution reactions at nitrogen (p K a of the leaving acid) and steric effects of para-aryl substituents ( E s). A number of mutagens exhibit activities that deviate markedly from the predicted values and they fall into two classes: di- tert-butylated N-benzoyloxy- N-benzyloxybenzamides, which – because of their size – are most probably excluded from the major groove or are unable to achieve a transition state for reaction with DNA, and N-benzoyloxy- N-butoxyalkylamides with branching α-to the amide carbonyl, which are resistant to S N2 reactions at the amide nitrogen.

Mutagenicity of Surface Soils in Urban Areas of Aichi Prefecture, Japan, and Bangkok, Thailand

To clarify the contamination levels of surface soil with mutagens in urban areas of Aichi prefecture, Japan, and Bangkok, Thailand, 60 and 67 surface soil samples were collected in Aichi and Bangkok, respectively, and mutagenicities of the organic extracts from these soil samples were examined in the Ames assay using Salmonella typhimurium (S. typhimurium) TA98 and TA100 with and without a mammalian metabolic system (S9 mix). Most of the soil samples collected in both areas showed mutagenicity in TA98 with and without S9 mix and in TA100 with S9 mix. Thirteen (22%) soil samples collected in Aichi showed high mutagenicity (more than 1000 revertants/g of soil) in TA98 with and/or without S9 mix, and six soil samples induced more than 4000 revertants/g of soil in TA98. Mutagenic potencies of most of the soil samples from Bangkok were moderate (100‐500 revertants/g of soil) or low (less than 100 revertants/g of soil) in both strains with and without S9 mix, and only one sample induced more than 1000 revertants/g of soil in both strains. In the characterization of mutagens, organic extracts of two and one soil samples from Aichi and Bangkok, respectively, were separated on an octadecylsilyl (ODS) column for HPLC, and mutagenicities of the resulting fractions were examined using S. typhimurium YG1024 without S9 mix. For the two soil samples from Aichi, retention times of the distinct mutagenic fractions were similar at 40‐74 min, and especially high mutagenicities were detected in three fractions corresponding to dinitropyrene isomers. In contrast, distinct mutagenicities were detected at retention times of 6‐29 min for the sample from Bangkok, and marked mutagenicity was found in the fraction corresponding to another nitroarene. These results indicate that surface soils in urban areas in Aichi and Bangkok are commonly contaminated with mutagens, but major mutagenic constituents in the surface soils are different.

Mutagens in contaminated soil: a review

The intentional and accidental discharges of toxic pollutants into the lithosphere results in soil contamination. In some cases (e.g., wood preserving wastes, coal-tar, airborne combustion by-products), the contaminated soil constitutes a genotoxic hazard. This work is a comprehensive review of published information on soil mutagenicity. In total, 1312 assessments of genotoxic activity from 118 works were examined. The majority of the assessments (37.6%) employed the Salmonella mutagenicity test with strains TA98 and/or TA100. An additional 37.6% of the assessments employed a variety of plant species (e.g., Tradescantia clone 4430, Vicia faba, Zea mays, Allium cepa) to assess mutagenic activity. The compiled data on Salmonella mutagenicity indicates significant differences (p<0.0001) in mean potency (revertents per gram dry weight) between industrial, urban, and rural/agricultural sites. Additional analyses showed significant empirical relationships between S9-activated TA98 mutagenicity and soil polycyclic aromatic hydrocarbon (PAH) concentration (r2=0.19 to 0.25, p<0.0001), and between direct-acting TA98 mutagenicity and soil dinitropyrene (DNP) concentration (r2=0.87, p<0.0001). The plant assay data revealed excellent response ranges and significant differences between heavily contaminated, industrial, rural/agricultural, and reference sites, for the anaphase aberration in Allium cepa (direct soil contact) and the waxy locus mutation assay in Zea mays (direct soil contact). The Tradescantia assays appeared to be less responsive, particularly for exposures to aqueous soil leachates. Additional data analyses showed empirical relationships between anaphase aberrations in Allium, or mutations in Arabidopsis, and the 137Cs contamination of soils. Induction of micronuclei in Tradescantia is significantly related to the soil concentration of several metals (e.g., Sb, Cu, Cr, As, Pb, Cd, Ni, Zn). Review of published remediation exercises showed effective removal of genotoxic petrochemical wastes within one year. Remediation of more refractory genotoxic material (e.g., explosives, creosote) frequently showed increases in mutagenic hazard that remained for extended periods. Despite substantial contamination and mutagenic hazards, the risk of adverse effect (e.g., mutation, cancer) in humans or terrestrial biota is difficult to quantify.

大気浮遊粒子，河川水および土砂の変異原性モニタリング―７年間（１９９６〜２００３年）の結果―

Airborne particles (n=541) , river waters (n=541) and surface soils (n=544) were collected from five geographically different regions of Japan (Hokkaido, Kanto, Chubu, Kinki and Kyushu regions) between 1996 and 2003. Extracts from those samples were measured and compared by a pre-incubation method using Salmonella typhimurium strains and a standard reference. Airborne particles were collected by high volume air samplers. All organic extracts of airborne particles except for some of those from Kinki region were mutagenic toward both TA98 and TA100 strains with and without S9 mix. Higher mutagenicity of airborne particulate extracts was observed for the samples collected at sites in heavy traffic areas and nearby industrial steel works. Mutagenic activity of airborne particles showed a gradual decrease over the period of the 7 years. Most of the blue rayon-absorbed materials from river waters exerted strong mutagenicity toward YG1024 strain with S9 mix, and the potencies varied widely among sampling sites. Blue rayonabsorbed materials showed no strong mutagenicity in TA100 or YG1029 strains, with or without S9 mix. Non-agricultural surface soils were collected from roadsides, parks and other open areas from each region. All organic extracts of surface soils were mutagenic toward either TA98 or TA100 strains with or without S9 mix. Mutagenic activities of surface soil extracts showed a wide range, and the effect of additional S9 mix on the activities depended on samples.