Mutation spectra of the drinking water mutagen 3-chloro-4-methyl-5-hydroxy-2(5H)-furanone (MCF) in Salmonella TA100 and TA104: comparison to MX

The chlorinated drinking water mutagen 3-chloro-4-methyl-5-hydroxy-2(5H)-furanone (MCF) occurs at concentrations similar to or greater than that of the related furanone 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX). MCF and MX differ structurally only by replacement of a 3-methyl in MCF with a 3-dichloromethyl in MX; yet, MCF is significantly less mutagenic than MX and produces different adducts when reacted with nucleosides or DNA. To explore further the effects that these structural differences might have on the biological activity of MCF and MX, we determined the mutation spectra of MCF in Salmonella strains TA100 and TA104 and of MX in strain TA104; the spectrum of MX in TA100 had been determined previously. In TA100, which presents only GC targets for mutagenesis, MCF induced primarily (75%) GC --> TA transversions, with most of the remaining revertants (20%) being GC --> AT transitions. This spectrum was not significantly different from that of MX in TA100 (P = 0.07). In TA104, which presents both GC and AT targets, MCF induced a lower percentage (57%) of GC --> TA transversions, with most of the remaining revertants (33%) being AT --> TA transversions. In contrast, MX induced almost only (98%) GC --> TA transversions in TA104, with the remaining revertants (2%) being AT --> TA transversions. Thus, almost all (98%) of the MX mutations were targeted at GC sites in TA104, whereas only 63% of the MCF mutations were so targeted. These results are consistent with the published findings that MX: (1) forms an adduct on guanosine when reacted with guanosine, (2) induces apurinic sites in DNA, and (3) forms a minor adduct on adenosine when reacted with adenosine or DNA. The results are also consistent with evidence that MCF forms adenosine adducts when reacted with adenosine. Our results show that the replacement of the 4-methyl in MCF with a 4-dichloromethyl to form MX not only increases dramatically the mutagenic potency but also shifts significantly the mutagenic specificity from almost equal targeting of GC and AT sites by MCF to almost exclusive targeting of GC sites by MX

Bioassay-directed fractionation and sub-fractionation for mutagenicity and chemical analysis of diesel exhaust particles

Several types of diesel exhaust particles (DEPs) have been used for toxicology studies, including a highorganic automobile DEP (A-DEP) from Japan, and a low-organic forklift DEP developed by the National Institute of Standards and Technology (N-DEP). However, these DEPs were not characterized extensively for chemical composition or sub-fractionated and tested extensively for mutagenicity. We collected a compressor-generated DEP (C-DEP) and characterized it by conducting bioassay-directed fractionation of the extractable organics in Salmonella and correlating the results by hierarchical clustering with the concentrations of 32 polycyclic aromatic hydrocarbons (PAHs). Relative to A- and N-DEP, the mutagenic potency of C-DEP was intermediate in TA100 1S9 (PAH mutagenicity) but was lowest in TA98 – S9 (nitroarene mutagenicity). More than 50% of the mass of the extractable organics of C-DEP eluted in the nonpolar fraction 1, and only -20% eluted in the moderately polar Fractions 2 and 3. However, most of the mutagenicity eluted in Fractions 2 and 3, similar to A-DEP but different from N-DEP. HPLC-derived mutagrams of 62 sub-fractions per fraction confirmed that most of the mutagenicity was due to moderately polar compounds. The diagnostic strains identified a strong role for PAHs, nitroarenes, aromatic amines, and oxy-PAHs in the mutagenicity of C-DEP. Hierarchical clustering confirmed the importance of oxy-PAHs but not that of nitroarenes. To our knowledge this is the first use of hierarchical clustering to correlate chemical composition with the mutagenicity of a complex mixture. The chemical analysis and mutagenicity of C-DEP described here makes C-DEP suitable for additional toxicological studies

Health effects of soy-biodiesel emissions: mutagenicity-emission factors.

CONTEXT: Soy biodiesel is the predominant biodiesel fuel used in the USA, but only a few, frequently conflicting studies have examined the potential health effects of its emissions

What's in the pool? a comprehensive identification of disinfection by-products and assessment of mutagenicity of chlorinated and brominated swimming pool water

Background: Swimming pool disinfectants and disinfection by-products (DBPs) have been linked to human health effects, including asthma and bladder cancer, but no studies have provided a comprehensive identification of DBPs in the water and related that to mutagenicity. Objectives: We performed a comprehensive identification of DBPs and disinfectant species in waters from public swimming pools in Barcelona, Catalonia, Spain, that disinfect with either chlorine or bromine and we determined the mutagenicity of the waters to compare with the analytical results. Methods: We used gas chromatography/mass spectrometry (GC/MS) to measure trihalomethanes in water, GC with electron capture detection for air, low- and high-resolution GC/MS to comprehensively identify DBPs, photometry to measure disinfectant species (free chlorine, monochloroamine, dichloramine, and trichloramine) in the waters, and an ion chromatography method to measure trichloramine in air. We assessed mutagenicity with the Salmonella mutagenicity assay. Results: We identified > 100 DBPs, including many nitrogen-containing DBPs that were likely formed from nitrogen-containing precursors from human inputs, such as urine, sweat, and skin cells. Many DBPs were new and have not been reported previously in either swimming pool or drinking waters. Bromoform levels were greater in brominated than in chlorinated pool waters, but we also identified many brominated DBPs in the chlorinated waters. The pool waters were mutagenic at levels similar to that of drinking water (~ 1,200 revertants/L-equivalents in strain TA100-S9 mix). Conclusions: This study identified many new DBPs not identified previously in swimming pool or drinking water and found that swimming pool waters are as mutagenic as typical drinking waters

Mutagenic activity of airborne particulate matter from the urban area of Porto Alegre, Brazil

The mutagenic activity of airborne particulate matter collected from three different sites within the urban area of Porto Alegre, Brazil, was investigated using a Salmonella/microsome assay. Samples were extracted by sonication, sequentially, with cyclohexane (CX), and dichloromethane (DCM), for a rough fractionation by polarity. The different fractions were tested for mutagenicity using Salmonella typhimurium strains TA98, with and without metabolic activation (S9 mix fraction), and TA98NR and TA98/1,8-DNP6, without metabolic activation. Mutagenic response was observed for frameshift strain TA98 in assays with and without metabolization for two sites (sites 2 and 3), which had considerable risk of environmental contamination by nonpolar (CX) and/or moderately polar (DCM) compounds. However, the values of revertants/m3 (rev/m3) were highest on the site subject to automobile exhaust (site 3) in assays without (9.56 rev/m3) and with metabolization (5.08 rev/m3). Maximum mutagenic activity was detected in the moderately polar fraction, decreasing after metabolization. Nevertheless, the nonpolar fractions (CX) gave higher mutagenic activity in the presence of metabolization than in the absence of the S9 mix fraction. The responses observed for TA98NR and TA98/1,8-DNP6 strains suggest the activity of nitrocompounds.<br>Foi investigada a atividade mutagênica de material particulado de amostras de ar coletadas em três diferentes locais dentro da área urbana da cidade de Porto Alegre, Brasil, através do ensaio Salmonella/microssoma. As amostras foram extraídas, em ultra-som, por fracionamento seqüencial de acordo com a polaridade, utilizando os solventes ciclohexano (CX) e diclorometano (DCM). As diferentes frações foram testadas para mutagenicidade com as linhagens de Salmonella typhimurium TA98, em presença e ausência de ativação metabólica, e TA98NR e TA98/1,8-DNP6 em ausência de metabolização. Observou-se resposta mutagênica positiva, do tipo erro no quadro de leitura, na linhagem TA98 (em ensaios em presença e ausência de metabolização) para compostos não polares (CX) e/ou moderadamente polares (DCM) nos locais com considerável risco de contaminação ambiental (locais 2 e 3). No entanto, os valores de revertentes por m3 (rev/m³) foram mais elevados no local sujeito a maior influência de veículos automotores (local 3), tanto em ensaios em ausência (9,56 rev/m³) como em presença de metabolização (5,08 rev/m³). A atividade mutagênica máxima foi detectada na fração moderadamente polar, decrescendo após metabolização. No entanto, as frações não polares (CX) apresentaram atividade mutagênica mais elevada na presença do que na ausência de fração S9mix. As respostas observadas para as linhagens TA98NR e TA98/1,8-DNP6 sugerem a contribuição de nitrocompostos na atividade mutagênica observada

Assessment of DNA damage using chromosomal aberrations assay in lymphocytes of waterpipe smokers

OBJECTIVES: The aim of this study was to investigate the genotoxicity of waterpipe smoking in the lymphocytes of waterpipe smokers using chromosomal aberrations (CAs) assay. MATERIALS AND METHODS: Fifty waterpipe smokers and 18 healthy non-smokers volunteered to participate in the study. Additionally, 18 heavy cigarette smokers were recruited for comparison. Chromosomal aberrations (CAs) assay was used to evaluate DNA damage in the lymphocytes. RESULTS: The results showed that similarly to cigarette smoking, waterpipe smoking significantly increased the frequencies of CAs (p < 0.01). In addition, the frequencies of CAs increased with more waterpipe use. CONCLUSIONS: Waterpipe smoking causes DNA damage to lymphocytes and the damage increases with more waterpipe use