RESUMEN
To establish the risk of the endocrine disrupting activity of polycyclic aromatic compounds, especially oxygenated and nitrated polycyclic aromatic hydrocarbons (oxy-PAHs and nitro-PAHs, respectively), antiandrogenic and estrogenic activities were determined using chemically activated luciferase expression (CALUX) assays with human osteoblast sarcoma cells. A total of 27 compounds including 9 oxy-PAHs (polycyclic aromatic ketones and quinones) and 8 nitro-PAHs was studied. The oxy-PAHs of 7H-benz[de]anthracen-7-one (BAO), 11H-benzo[a]fluoren-11-one (B[a]FO), 11H-benzo[b]fluoren-11-one (B[b]FO), and phenanthrenequinone (PhQ) exhibited significantly the potent inhibition of AR activation. All nitro-PAHs exhibited high antiandrogenic activities (especially high for 3-nitrofluoranthene (3-NFA) and 3-nitro-7H-benz[de]anthracen-7-one (3-NBAO)), and the AR inhibition was confirmed as noncompetitive for 3-NFA, 3-NBAO, and 1,3-dinitropyrene (1,3-DNPy). Antiandrogenic activity of 3-NFA demonstrated characteristically a U-shaped dose-response curve; however, the absence of fluorescence effect on the activity was confirmed. The prominent estrogenic activity dependent on dose-response curve was confirmed for 2 oxy-PAHs (i.e., B[a]FO and B[b]FO). Elucidating the role of AR and ER on the effects of polycyclic aromatic compounds (e.g., oxy- and nitro-PAHs) to endocrine dysfunctions in mammals and aquatic organisms remains a challenge.
Asunto(s)
Hidrocarburos Policíclicos Aromáticos , Compuestos Policíclicos , Animales , Humanos , Hidrocarburos Policíclicos Aromáticos/toxicidad , Hidrocarburos Policíclicos Aromáticos/química , Nitratos/química , Quinonas , Luciferasas , MamíferosRESUMEN
The question of what kinds of airborne particles, including diesel exhaust particles and their adherent chemical constituents, exacerbate the activity of allergic and inflammatory respiratory diseases has not been elucidated in detail. Therefore, chemicals that have amplifying actions on Dermatophagoides farinae (Df) body extract-induced IL-8, the inflammatory cytokines of the innate immune system, were comprehensively examined using commonly used human alveolar epithelial cells, A549, as simple screening for 17 polycyclic aromatic hydrocarbons (PAHs), which are representative organic constituents in atmospheric samples. The significant amplifying actions of two PAHs, dibenzo[a,l]pyrene (DB[a,l]P) at 50 nM and dibenzo[a,i]pyrene (DB[a,i]P) at 2 µM for 48 h, for IL-8 protein release induced by mite antigens in epithelial cells were observed for the first time. In contrast, the enhancement of IL-8 was not observed in protein levels for these PAHs without the antigens. Meanwhile, the significant synergistic amplifying effect of DB[a,l]P at 50 nM on proinflammatory actions was measured in gene expression (i.e., IL-8, IL-6, ICAM-1, and TNF-α) levels in the experimental setting; for the results, the induction of TNF-α may have been the essential factor that enhanced the amplifying activity of DB[a,l]P for IL-8 gene expression and protein release. Examining the exacerbating effect on allergic pathophysiological states for DB[a,l]P is planned for further study.
RESUMEN
BACKGROUND: Conflicting results between bacterial mutagenicity tests (the Ames test) and mammalian carcinogenicity tests might be due to species differences in metabolism, genome structure, and DNA repair systems. Mutagenicity assays using human cells are thought to be an advantage as follow-up studies for positive results in Ames tests. In this collaborative study, a thymidine kinase gene mutation study (TK6 assay) using human lymphoblastoid TK6 cells, established in OECD TG490, was used to examine 10 chemicals that have conflicting results in mutagenicity studies (a positive Ames test and a negative result in rodent carcinogenicity studies). RESULTS: Two of 10 test substances were negative in the overall judgment (20% effective as a follow-up test). Three of these eight positive substances were negative after the short-term treatment and positive after the 24 h treatment, despite identical treatment conditions without S9. A toxicoproteomic analysis of TK6 cells treated with 4-nitroanthranilic acid was thus used to aid the interpretation of the test results. This analysis using differentially expressed proteins after the 24 h treatment indicated that in vitro specific oxidative stress is involved in false positive response in the TK6 assay. CONCLUSIONS: The usefulness of the TK6 assay, by current methods that have not been combined with new technologies such as proteomics, was found to be limited as a follow-up test, although it still may help to reduce some false positive results (20%) in Ames tests. Thus, the combination analysis with toxicoproteomics may be useful for interpreting false positive results raised by 24 h specific reactions in the assay, resulting in the more reduction (> 20%) of false positives in Ames test.
RESUMEN
Studies have been conducted on the genotoxicity and carcinogenicity of disinfection by-products formed from natural organic matter (NOM) and mitigation effect for mutagens and clastogens by NOM. Whereas, reportedly, synthetic humic acid in high concentration has induced genotoxicity in human cells, and NOM samples have provoked mild oxidative and other physiological responses in aquatic organisms. Our group developed a novel detection method for DNA damage formation, namely enhanced green fluorescent protein (EGFP)-fused mediator of DNA damage checkpoint 1 (MDC1)-expressing human cells as simple and high-sensitive system. By using this method, a significant increase in the foci area was observed after 3â¯h, but not 24â¯h for 130 mgC L-1 Suwannee River fulvic acid (SRFA), 38 mgC L-1 humic acid (SRHA), and 19 mgC L-1 NOM (SRNOM). The SRNOM concentration is the original environmental one; therefore, it was suggested that the formation and repair of DNA damage associated with γ-H2AX, a biomarker for DNA double-strand breaks by mild oxidative stress, in Suwannee River (SR) were detected for the first time. The increase in the foci area was not observed for 18 mgC L-1 Lake Biwa fulvic acid (LBFA) and 50â¯mgâ¯L-1 catechin after both 3â¯h and 24â¯h. The difference between the SR and Lake Biwa (LB) samples may result from the differences in their electron-accepting capacity. The application of this methodology is expected to elucidate oxidative stress and toxicological effects shortly and in detail for many water samples.
Asunto(s)
Benzopiranos/toxicidad , Daño del ADN/efectos de los fármacos , Daño del ADN/genética , Proteínas Fluorescentes Verdes/genética , Sustancias Húmicas/toxicidad , Proteínas Nucleares/biosíntesis , Transactivadores/biosíntesis , Proteínas Adaptadoras Transductoras de Señales , Proteínas de Ciclo Celular , Línea Celular Tumoral , Colorantes , Humanos , Lagos/química , Células MCF-7 , Proteínas Nucleares/genética , Ríos/química , Transactivadores/genéticaRESUMEN
The effect of waterborne ingredient on ecosystem has been of great interest. In the present study, the evaluation method using algal photosynthesis inhibition assay with dual-channel pulse amplitude modulation (PAM) system was established for a series of water samples to elucidate the potential effect of the total body of organic compounds including natural organic matter (NOM) on aquatic ecosystems. The more sensitive and less time-consuming monitoring method compared with algal growth inhibition assay was suggested, especially considering inorganic and coloring constituents. Algal photosynthesis inhibition activity was detected with high sensitivity for photosystem II (PSII) inhibitors, whereas the IC10 of the other chemicals was over the environmental standard concentration for Chlamydomonas moewusii (Chlorophyceae) and Pheodactylum tricornutum (Diatomea). The photosynthesis inhibition activity of Lake Biwa dissolved organic matter (LBDOM) and fulvic acid (LBFA) was significantly detected at ≥10 times the concentration and >10â¯mgC L-1, respectively, whereas prominent activity was confirmed for Suwannee River NOM (SRNOM) on the river original concentration (>30â¯mgC L-1) for both algae. Significant inhibition activity was detected in both algae at least in twice-concentration for water samples from a wastewater treatment pilot plant. There was no great difference in the activity between sewage secondary effluent and its filtrate with ultrafiltration (UF), and physically washing water for the UF membrane.
Asunto(s)
Chlamydomonas/efectos de los fármacos , Diatomeas/efectos de los fármacos , Sustancias Húmicas/análisis , Fotosíntesis/efectos de los fármacos , Contaminantes Químicos del Agua/análisis , Purificación del Agua/métodos , Compuestos de Amonio/análisis , Chlamydomonas/fisiología , Colorantes/análisis , Diatomeas/fisiología , Lagos/química , Ultrafiltración/métodos , Aguas Residuales/química , Contaminantes Químicos del Agua/toxicidadRESUMEN
Various types of polycyclic aromatic compounds (PACs) in diesel exhaust particles are thought to contribute to carcinogenesis in mammals. Although the carcinogenicity, mutagenicity and tumour-initiating activity of these compounds have been evaluated, their tumour-promoting activity is unclear. In the present study, to determine the tumour-inducing activity of PACs, including previously known mutagenic compounds in atmospheric environments, a transformation assay for promoting activity mediated by the release of contact inhibition was conducted for six polycyclic aromatic hydrocarbons (PAHs), seven oxygenated PAHs (oxy-PAHs) and seven nitrated PAHs (nitro-PAHs) using mouse embryonic fibroblast cells transfected with the v-Ha-ras gene (Bhas 42 cells). Of these, two PAHs [benzo[k]fluoranthene (B[k]FA) and benzo[b]fluoranthene (B[b]FA)], one oxy-PAH [6H-benzo[cd]pyren-6-one (BPO)] and two nitro-PAHs (3-nitro-7H-benz[de]anthracen-7-one and 6-nitrochrysene) were found to exhibit particularly powerful tumour-promoting activity (≥10 foci following exposure to <100nM). In addition, clear mRNA expression of CYP1A1, which is associated with aryl hydrocarbon receptor (AhR)-mediated activation, was observed following the exposure of cells to two PAHs (B[k]FA and B[b]FA) and three oxy-PAHs (1,2-naphthoquinone, 11H-benzo[b]fluoren-11-one and BPO). Further, an HO-1 antioxidant response activation was observed following exposure to B[k]FA, B[b]FA and BPO, suggesting that the induction of tumour-promoting activity in these compounds is correlated with the dysfunction of signal transduction via AhR-mediated responses and/or oxidative stress responses.
Asunto(s)
Carcinógenos/química , Carcinógenos/toxicidad , Transformación Celular Neoplásica/inducido químicamente , Hidrocarburos Policíclicos Aromáticos/química , Hidrocarburos Policíclicos Aromáticos/toxicidad , Animales , Línea Celular , Supervivencia Celular/efectos de los fármacos , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Ratones , Mutágenos/toxicidad , ARN Mensajero/genética , Emisiones de Vehículos/toxicidadRESUMEN
Sulfuric acid-treated liver extracts of representative high-trophic level Japanese animals were analyzed by toxic identification and evaluation (TIE) with chemically activated luciferase expression (CALUX) and chemical analysis to elucidate androgen receptor (AR) antagonistic activities and potential contributions of organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs). The activities were detected in striped dolphins (n = 5), Stejneger's beaked whales (n = 6), golden eagle (n = 1), and Steller's sea eagle (n = 1) with CALUX-flutamide equivalents (FluEQs) as follow: 38 (20-52), 47 (21-96), 5.0, and 80 µg FluEQ/g-lipid, respectively. The AR antagonism was detected in limited number of specimens at lower levels for finless porpoise, raccoon dog, and common cormorant. Theoretical activities (Theo-FluEQs) were calculated using the concentration of OCPs and PCBs and their IC25-based relative potency (REP) values. These total contribution to CALUX-FluEQ was 126%, 84%, 53%, 55%, and 44% for striped dolphin, Steller's sea eagle, Stejneger's beaked whale, finless porpoise, and golden eagle, respectively, and the main contributor was p,p'-DDE. However, most of the activities for raccoon dog (7.6%) and common cormorant (17%) could not be explained by OCPs and PCBs. This suggests other unknown compounds could function as AR antagonists in these terrestrial species.
Asunto(s)
Antagonistas de Receptores Androgénicos/análisis , Ecotoxicología/métodos , Extractos Hepáticos/análisis , Plaguicidas/análisis , Antagonistas de Receptores Androgénicos/metabolismo , Animales , Animales Salvajes/metabolismo , Aves , Diclorodifenil Dicloroetileno/análisis , Águilas , Monitoreo del Ambiente/métodos , Cadena Alimentaria , Hidrocarburos Clorados/análisis , Hidrocarburos Clorados/toxicidad , Japón , Extractos Hepáticos/metabolismo , Plaguicidas/toxicidad , Bifenilos Policlorados/análisis , Bifenilos Policlorados/toxicidad , Marsopas , Perros Mapache , Receptores Androgénicos/metabolismo , Contaminantes Químicos del Agua/análisis , Contaminantes Químicos del Agua/metabolismo , Contaminantes Químicos del Agua/toxicidad , Ballenas/metabolismoRESUMEN
Dioxin-Responsive Chemical-Activated LUciferase gene eXpression assay (DR-CALUX) was applied to assess the total toxic activity of the mixture of PAHs and related compounds as well as dioxin-related compounds in road dust from urban areas of Hanoi, Vietnam. Road dust from Hanoi contained significantly higher DR-CALUX activities (3 to 39, mean 20 ng CALUX-TEQ/g dw) than those from a rural site (2 to 13, mean 5 ng CALUX-TEQ/g dw). The total concentrations of 24 major PAHs (Σ24PAHs) in urban road dust (0.1 to 5.5, mean 2.5 µg/g dw) were also 6 times higher than those in rural road dust (0.08 to 1.5, mean 0.4 µg/g dw). Diagnostic ratios of PAHs indicated vehicular engine combustion as the major PAH emission source in both sites. PAHs accounted for 0.8 to 60% (mean 10%) and 2 to 76% (mean 20%) of the measured CALUX-TEQs in road dust for Hanoi the rural site, respectively. Benzo[b]-/benzo[k]fluoranthenes were the major TEQ contributors among PAHs, whereas DRCs contributed <0.1% to CALUX-TEQs for both rural and urban sites. These results suggest TEQ contribution of other aryl hydrocarbon receptor agonists in road dust. Significant PAH concentrations in urban dust indicated high mutagenic and carcinogenic potencies. Estimated results of incremental life time cancer risk (ILCR) indicated that Vietnamese populations, especially those in urban areas such as Hanoi, are potentially exposed to high cancer risk via both dust ingestion and dermal contact. This is the first study on the exposure risk of AhR agonists, including PAHs and DRCs, in urban road dust from a developing country using a combined bio-chemical analytical approach.
Asunto(s)
Polvo/análisis , Exposición a Riesgos Ambientales/estadística & datos numéricos , Contaminantes Ambientales/toxicidad , Hidrocarburos Policíclicos Aromáticos/toxicidad , Receptores de Hidrocarburo de Aril/metabolismo , Bioensayo , Exposición a Riesgos Ambientales/análisis , Monitoreo del Ambiente , Contaminantes Ambientales/análisis , Humanos , Hidrocarburos Policíclicos Aromáticos/análisis , Medición de Riesgo , VietnamRESUMEN
There are very few reports on the contamination by perfluorinated chemicals (PFCs) in the environment of developing countries, especially regarding their emission from waste recycling and disposal sites. This is the first study on the occurrence of a wide range of PFCs (17 compounds) in ambient water in Vietnam, including samples collected from a municipal dumping site (MD), an e-waste recycling site (ER), a battery recycling site (BR) and a rural control site. The highest PFC concentration was found in a leachate sample from MD (360 ng/L). The PFC concentrations in ER and BR (mean, 57 and 16 ng/L, respectively) were also significantly higher than those detected in the rural control site (mean, 9.4 ng/L), suggesting that municipal solid waste and waste electrical and electronic equipment are potential contamination sources of PFCs in Vietnam. In general, the most abundant PFCs were perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), and perfluoroundecanoic acid (PFUDA; <1.4-100, <1.2-100, and <0.5-20 ng/L, respectively). Interestingly, there were specific PFC profiles: perfluoroheptanoic acid and perfluorohexanoic acid (88 and 77 ng/L, respectively) were almost as abundant as PFOA in MD leachate (100 ng/L), whereas PFNA was prevalent in ER and BR (mean, 17 and 6.2 ng/L, respectively) and PFUDA was the most abundant in municipal wastewater (mean, 5.6 ng/L), indicating differences in PFC contents in different waste materials.
Asunto(s)
Fluorocarburos/análisis , Administración de Residuos/métodos , Contaminantes Químicos del Agua/análisis , Residuos Electrónicos , Monitoreo del Ambiente , Vietnam , Contaminación Química del Agua/estadística & datos numéricosRESUMEN
Diacylglycerol kinase (DGK) plays an important role in phosphoinositide signaling cascade by regulating the intracellular level of diacylglycerol and phosphatidic acid. The DGK family is involved in various pathophysiological responses that are mediated through unique binding partners in different tissues and cells. In this study, we identified a small GTPase effector protein, IQGAP1, as a novel DGKζ-associated complex protein. A bacterial endotoxin, lipopolysaccharide (LPS), facilitated the complex formation in macrophages. Both proteins co-localized at the edge and phagocytic cup of the cell. Furthermore, RNA interference-mediated knockdown of DGKζ or IQGAP1 impaired LPS-induced Rac1 activation. Primary macrophages derived from DGKζ(-/-) mice attenuated LPS-induced phagocytosis of bacteria. These results suggest that DGKζ is involved in IQGAP1/Rac1-mediated phagocytosis upon LPS stimulation in macrophages.
Asunto(s)
Diacilglicerol Quinasa/metabolismo , Macrófagos/inmunología , Neuropéptidos/metabolismo , Fagocitosis , Proteínas de Unión al GTP rac/metabolismo , Proteínas Activadoras de ras GTPasa/metabolismo , Animales , Células Cultivadas , Diacilglicerol Quinasa/genética , Humanos , Lipopolisacáridos/inmunología , Activación de Macrófagos , Ratones , Ratones Endogámicos C57BL , Proteína de Unión al GTP rac1RESUMEN
1H-Phenalen-1-one (phenalenone) is one of the major oxygenated polyaromatic compounds present in the atmospheric environment. In order to gain detailed information regarding the mutagenicity and physicochemical properties of the nitration products of phenalenone, we measured Ames Salmonella mutagenicity, lower LUMO (lowest unoccupied molecular orbital) energy and octanol-water partition coefficient of the products obtained from the nitration reaction of phenalenone. Both nitration reactions of phenalenone, i.e. with mixed inorganic acids (a mixture of nitric acid and sulphuric acid) and with NO(2)-O(3) in an aprotic solvent, preferentially afforded the nitration products 2-nitrophenalenone and 5-nitrophenalenone. Formation of a 6-nitro derivative of phenalenone was, however, only observed in the nitration reaction with sulphuric acid. Moreover, dinitro derivatives of phenalenone and also two oxidatively decomposed products of nitrophenalenone, i.e. 3-nitro- and 4-nitronaphthalic anhydride, were isolated from the reaction mixture. The mutagenicities of the six nitro compounds obtained from the nitration reactions were tested with the Salmonella strains TA98, TA100, YG1021 and YG1024 in the absence of S9 mix. Among these products, 2-nitrophenalenone exhibited the most potent mutagenic activity against TA98, TA100 and YG1024 (160, 230 and 2800 revertants/nmol for strains TA100, TA98 and YG1024, respectively), whereas 2,5-dinitrophenalenone exerted the highest mutagenicity against YG1021. Semi-empirical calculation showed that among the mononitrophenalenone series, the mononitro derivatives possessing lower LUMO energy tended to exhibit greater mutagenic activity than those with higher LUMO energy. This tendency, however, did not extend to the compounds with different aromatic ring systems due to the considerable differences in the hydrophobicities of these compounds.
Asunto(s)
Daño del ADN , Mutágenos/toxicidad , Nitrocompuestos/toxicidad , Fenalenos/química , Interacciones Hidrofóbicas e Hidrofílicas , Mutagénesis , Pruebas de Mutagenicidad , Mutágenos/química , Nitrocompuestos/química , Salmonella typhimurium/efectos de los fármacos , Salmonella typhimurium/genéticaRESUMEN
A wide variety of contaminants derived from diesel and gasoline engines, tire, asphalt, and natural organic compounds is found in road dust. Polycyclic aromatic compounds (PACs) are the important toxic targets among various contents in road dust and diesel exhaust particulates (DEPs), and endocrine-disrupting activity of PACs was suggested. In the present study, aryl hydrocarbon receptor (AhR) ligand activity was confirmed in the extract of both road dust and DEPs. In the separation of the extracts for both road dust and DEPs with reversed-phase HPLC, it was found that polar fractions contributed to significant AhR ligand activity in both a mouse hepatoma (H1L1) cell system and a yeast system. Furthermore, the contribution of these polar fractions was higher in DEPs than in road dust, probably because of the greater concentration of oxy-PAHs in DEPs than in road dust. The contribution of contaminants associated with the polar region to AhR ligand activity was also evident following the separation of road dust with normal-phase HPLC. Additionally, remarkable estrogen receptor (ER) ligand activity was detected in the highly polar region separated with normal-phase HPLC. It is suggested that many unknown AhR or ER ligand active compounds are contained in the polar region.
Asunto(s)
Contaminantes Atmosféricos/análisis , Material Particulado/análisis , Receptores de Hidrocarburo de Aril/metabolismo , Receptores de Estrógenos/metabolismo , Emisiones de Vehículos/análisis , Contaminantes Atmosféricos/toxicidad , Animales , Cromatografía Líquida de Alta Presión , Polvo/análisis , Japón , Ligandos , Ratones , Material Particulado/toxicidad , Hidrocarburos Policíclicos Aromáticos/análisis , Hidrocarburos Policíclicos Aromáticos/toxicidad , Saccharomyces cerevisiae/metabolismo , Células Tumorales Cultivadas , Emisiones de Vehículos/toxicidadRESUMEN
Polycyclic aromatic ketones (PAKs) and polycyclic aromatic quinones (PAQs) are oxygenated polycyclic aromatic hydrocarbons (PAHs), and reports about the aryl hydrocarbon receptor (AhR) ligand activities of these compounds are few. In this study, activation of AhR by 41 polycyclic aromatic compounds (PACs), focusing especially on PAKs and PAQs, was determined by measuring beta-galactosidase activity from a reporter plasmid in yeast engineered to express human AhR and the AhR nuclear translocator proteins and by measuring luciferase activity from mouse hepatoma (H1L1) cells (chemical-activated luciferase expression [CALUX] assay). The PACs used in these experiments included 11 PAKs, seven PAQs, and 21 PAHs. In this study, the PAKs 11H-benzo[a]fluoren-11-one (B[a]FO), 11H-benzo[b]fluoren-11-one (B[b]FO) and 7H-benzo[c]fluoren-7-one and the PAQs 5,12-naphthacenequinone, 1,4-chrysenequinone, and 7,12-benz[a]anthracenequinone showed high AhR activities in H1L1 cells, although these values were not as high as that for benzo[a]pyrene (B[a]P). These PAKs and PAQs showed significantly stronger activities in yeast cells relative to B[a]P. It was predicted that PAKs such as B[a]FO and B[b]FO occupied 0.06% to 1.3% of the total induction equivalents, and each contribution matched the contribution of PAHs such as B[a]P, chrysene, and benz[a]anthracene in gasoline exhaust particulates and airborne particulates using data of CALUX assay.
Asunto(s)
Cetonas/farmacología , Compuestos Policíclicos/farmacología , Quinonas/farmacología , Receptores de Hidrocarburo de Aril/metabolismo , Animales , Línea Celular , Ligandos , RatonesRESUMEN
N2-ethylidene-2'-deoxyguanosine (N2-ethylidene-dG) is a major DNA adduct induced by acetaldehyde. Although it is unstable in the nucleoside form, it is relatively stable when present in DNA. In this study, we analyzed three acetaldehyde-derived DNA adducts, N2-ethylidene-dG, N2-ethyl-2'-deoxyguanosine (N2-Et-dG) and alpha-methyl-gamma-hydroxy-1,N2-propano-2'-deoxyguanosine (alpha-Me-gamma-OH-PdG) in the liver DNA of aldehyde dehydrogenase (Aldh)-2-knockout mice to determine the influence of alcohol consumption and the Aldh2 genotype on the levels of DNA damage. In control Aldh2+/+ mice, the level of N2-ethylidene-dG adduct in liver DNA was 1.9 +/- 0.7 adducts per 10(7) bases and was not significantly different than that of Aldh2+/- and -/- mice. In alcohol-fed mice (20% ethanol for 5 weeks), the adduct levels of Aldh2+/+, +/- and -/- mice were 7.9 +/- 1.8, 23.3 +/- 4.0 and 79.9 +/- 14.2 adducts per 10(7) bases, respectively, and indicated that adduct level was alcohol and Aldh2 genotype dependent. In contrast, an alcohol- or Aldh2 genotype-dependent increase was not observed for alpha-Me-gamma-OH-PdG, and N2-Et-dG was not detected in any of the analyzed samples. In conclusion, the risk of formation of N2-ethylidene-dG in model animal liver in vivo is significantly higher in the Aldh2-deficient population and these results may contribute to our understanding of in vivo adduct formation in humans.
Asunto(s)
Aldehído Deshidrogenasa/metabolismo , Aductos de ADN/biosíntesis , Desoxiguanosina/análogos & derivados , Etanol/toxicidad , Hígado/efectos de los fármacos , Aldehído Deshidrogenasa/genética , Aldehído Deshidrogenasa Mitocondrial , Animales , Cromatografía Liquida , Aductos de ADN/química , Desoxiguanosina/química , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Espectrometría de Masas en TándemRESUMEN
Dyeing wastewater collected in Kyoto city, Japan, was investigated for the occurrence of aryl hydrocarbon receptor (AhR) ligands by using an AhR-responsive reporter gene assay. Concentrated extracts of wastewater samples elicited a dose-dependent increase in AhR ligand activity, and several hydrophobic HPLC fractions of the extracts were highly effective in inducing AhR ligand activity. Three potential AhR ligands were isolated from these fractions and identified to be Disperse Red 92, Disperse Yellow 64, and 3'-hydroxybenzo[b]quinophthalone by using HPLC and LC-MS/MS. Disperse Red 92, which has also been detected in the treated effluent from a sewage plant receiving the wastewater, is an anthraquinone disperse dye showing weak AhR binding affinity in the assay. Disperse Yellow 64 and 3'-hydroxybenzo[b]quinophthalone are quinoline disperse dyes capable of activating the AhR at nanomolar concentrations. In particular, Disperse Yellow 64 is a highly potent AhR ligand that was 3 times more effective in inducing AhR ligand activity than beta-naphthoflavone in the assay. Quinoline disperse dyes are suggested to be a new class of xenobiotic AhR ligands which pose a danger to aquatic biota and human health.
Asunto(s)
Compuestos Azo/aislamiento & purificación , Colorantes , Receptores de Hidrocarburo de Aril/metabolismo , Contaminantes Químicos del Agua/aislamiento & purificación , Xenobióticos/aislamiento & purificación , Compuestos Azo/análisis , Compuestos Azo/toxicidad , Cromatografía Líquida de Alta Presión , Relación Dosis-Respuesta a Droga , Genes Reporteros/genética , Ligandos , Estructura Molecular , Espectrometría de Masas en Tándem , Contaminantes Químicos del Agua/análisis , Contaminantes Químicos del Agua/toxicidad , Xenobióticos/análisis , Xenobióticos/toxicidad , LevadurasRESUMEN
Oxygenated polycyclic aromatic hydrocarbons (oxy-PAHs) such as polycyclic aromatic quinones and polycyclic aromatic ketones as well as polycyclic aromatic hydrocarbons (PAHs) are abundant in the atmospheric environment. In this study, mRNA induction of six metabolic enzymes including P4501A1, 1A2, and 1B1, aldo-keto reductase 1C1 (AKR1C1), NAD(P)H-dependent quinone oxidoreductase 1 (NQO1), and glutathione S-transferase M1 (GSTM1) were examined in detail in human hepatoma (HepG2) cells exposed to environmentally relevant 13 PAHs and seven oxy-PAHs. Most PAHs such as benzo[a]pyrene (B[a]P) showed significant induction of P4501A1 and 1A2 mRNA, while induction by oxy-PAHs such as 5,12-naphthacenequinone (NCQ) and 11H-benzo[b]fluoren-11-one (B[b]FO) occurred less strongly. AKR1C1 mRNA was significantly induced by oxy-PAHs, 11H-benzo[a]fluoren-11-one (B[a]FO), NCQ, cyclopenta[cd]pyren-3(4H)-one (CPPO), and B[b]FO and also by P450s-inducing PAHs such as B[a]P, benzo[k]fluoranthene (B[k]FA), and dibenz[a,h]anthracene (DB[a,h]A). Both chemical-dependent and time-dependent induction patterns of NQO1 mRNA were of the mixed types of P4501A1 and AKR1C1. The tendency for the decrease of GSTM1 mRNA was observed when exposed to PAHs B[a]P and B[k]FA.
Asunto(s)
20-Hidroxiesteroide Deshidrogenasas/efectos de los fármacos , Carcinoma Hepatocelular/metabolismo , Sistema Enzimático del Citocromo P-450/efectos de los fármacos , Glutatión Transferasa/efectos de los fármacos , NAD(P)H Deshidrogenasa (Quinona)/efectos de los fármacos , Hidrocarburos Policíclicos Aromáticos/farmacocinética , 20-Hidroxiesteroide Deshidrogenasas/metabolismo , Hidrocarburo de Aril Hidroxilasas/efectos de los fármacos , Hidrocarburo de Aril Hidroxilasas/metabolismo , Biotransformación/efectos de los fármacos , Línea Celular Tumoral , Citocromo P-450 CYP1A1/efectos de los fármacos , Citocromo P-450 CYP1A1/metabolismo , Citocromo P-450 CYP1A2/efectos de los fármacos , Citocromo P-450 CYP1A2/metabolismo , Citocromo P-450 CYP1B1 , Sistema Enzimático del Citocromo P-450/metabolismo , Relación Dosis-Respuesta a Droga , Inducción Enzimática/efectos de los fármacos , Glutatión Transferasa/metabolismo , Humanos , Estructura Molecular , NAD(P)H Deshidrogenasa (Quinona)/metabolismo , Hidrocarburos Policíclicos Aromáticos/metabolismo , ARN Mensajero/efectos de los fármacos , ARN Mensajero/metabolismo , Relación Estructura-Actividad , Factores de TiempoRESUMEN
The mechanism of Preferential Enrichment, an unusual enantiomeric resolution phenomenon observed upon recrystallization of a series of racemic crystals which are classified as a racemic mixed crystal with fairly ordered arrangement of the two enantiomers, has been studied. On the basis of the existence of polymorphs and the occurrence of the resulting polymorphic transition during crystallization from solution, the mechanism has been accounted for in terms of (1) a preferential homochiral molecular association to form one-dimensional chain structures in the supersaturated solution of the racemate or nonracemic sample with a low ee value, (2) a kinetic formation of a metastable crystalline phase retaining the homochiral chain structures in a process of nucleation, (3) a polymorphic transition from the metastable phase to a stable one followed by enantioselective liberation of the excess R (or S) enantiomers from the transformed crystal into solution at the beginning of crystal growth to result in a slight enrichment (up to 10% ee) of the opposite S (or R) enantiomer in the deposited crystals, together with an enantiomeric enrichment of the R (or S) enantiomer in the mother liquor, and (4) a chiral discrimination by the once formed S (or R)-rich stable crystalline phase in a process of the subsequent crystal growth, leading to a considerable enantiomeric enrichment of the R (or S) enantiomer up to 100% ee in the mother liquor. The processes (3) and (4) are considered to be directly responsible for an enrichment of one enantiomer in the mother liquor. The association mode of the two enantiomers in solution has been investigated by means of (i) the solubility measurement and (ii) the number-averaged molecular weight measurement in solution by vapor pressure osmometry, together with (iii) the molecular dynamics simulation of oligomer models. The polymorphic transition during crystallization has been observed visually and by means of the in situ FTIR technique and DSC measurement. Both metastable and stable crystals have been obtained, and their crystal structures have been elucidated by X-ray crystallographic analysis of their single crystals.