• BMB Reports
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• 제41권8호
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• pp.604-608
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• 2008

1-Nitropyrene 4,5-oxide and 1-nitropyrene 9,10-oxide are oxidative metabolites that are responsible for the mutagenicity of 1-nitropyrene. In this study, the mutation spectra induced by oxidative metabolites in human cells were determined using a shuttle vector assay. The mutation frequencies induced by 1-nitropyrene 9,10-oxide were 2-3 times higher than those induced by 1-nitropyrene 4,5-oxide. The base substitutions induced by 1-nitropyrene 4,5-oxide were $G{\rightarrow}A$ transitions, $G{\rightarrow}C$ transversions, and $G{\rightarrow}T$ transversions. In the case of 1-nitropyrene 9,10-oxide, $G{\rightarrow}A$ transitions, $G{\rightarrow}T$ transversions, $A{\rightarrow}G$ transitions and $G{\rightarrow}C$ transversions were observed. Most base substitution mutations induced by oxidative metabolites occurred at the guanine sites in the supF gene. These sequence-specific hot spots were commonly identified as 5'-GA sequences for both metabolites. On the other hand, the sequence-specific hot spots at the adenine sites were identified as 5'-CAC sequences for 1-nitropyrene 9,10-oxide. These results suggest that the oxidative metabolites of 1-nitropyrene induce sequence-specific DNA mutations at the guanine and adenine sites at high frequency.

• Molecules and Cells
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• 제19권1호
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• pp.114-123
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• 2005

Nitropyrene, the predominant nitropolycyclic hydrocarbon found in diesel exhaust, is a mutagenic and tumorigenic environmental pollutant that requires metabolic activation via nitroreduction and ring oxidation. In order to determine the role of ring oxidation in the mutagenicity of 1-nitropyrene, its oxidative metabolites, 1-nitropyrene 4,5-oxide and 1-nitropyrene 9,10-oxide, were synthesized and their mutation spectra were determined in the coding region of hprt gene of CHO cells by a PCR amplification of reverse-transcribed hprt mRNA, followed by a DNA sequence analysis. A comparison of the two metabolites for mutation frequencies showed that 1-nitropyrene 9,10-oxide was 2-times higher than 1-nitropyrene 4,5-oxide. The mutation spectrum for 1-nitropyrene 4,5-oxide was base substitutions (33/49), one base deletions (11/49) and exon deletions (5/49). In the case of 1-nitropyrene 9,10-oxide, base substitutions (27/50), one base deletions (15/50), and exon deletions (8/50) were observed. Base substitutions were distributed randomly throughout the hprt gene. The majority of the base substitutions in mutant from 1-nitropyrene 4,5-oxide treated cells were $A{\rightarrow}G$ transition (15/33) and $G{\rightarrow}A$ transition (8/33). The predominant base substitution, $A{\rightarrow}G$ transition (11/27) and $G{\rightarrow}A$ transition (8/27), were also observed in mutant from 1-nitropyrene 9,10-oxide treated cells. The mutation at the site of adenine and guanine was consistent with the previous results, where the sites of DNA adduct formed by these compounds were predominant at the sites of purines. A comparison of the mutational patterns between 1-nitropyrene 4,5-oxide and 1-nitropyrene 9,10-oxide showed that there were no significant differences in the overall mutational spectrum. These results indicate that each oxidative metabolite exhibits an equal contribution to the mutagenicity of 1-nitropyrene, and ring oxidation of 1-nitropyrene is an important metabolic pathway to the formation of significant lethal DNA lesions.

• 한국환경보건학회지
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• 제17권2호
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• pp.114-120
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• 1991

대기부유입자상물질이나 diesel 배출가스중에 함유하여, 주요한 direct-acting mutagen의 하나로 작용하는 1-nitropyrene, 1,3-, 1,6- 및 1,8-dinitropyrene을 합성하고, 고속액체 chromatography로 분리정제하여, Salmonella typhimurium TA 98, S9mix 비첨가의 계에서 돌연변이원성을 측정한 결과, 1-nitropyrene에 비하여, dinitropyrene은 강력한 돌연변이 원성을 나타내었고, 그 중에서도 1,8-dinitropyrene은 733,000 revertants/$\mu$g으로 최고의 돌연변이원 활성을 나타내었다.

• Journal of Microbiology and Biotechnology
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• 제32권7호
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• pp.869-876
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• 2022

The skin, which is the largest organ of the human body, is in direct contact with pollutants in the surrounding atmosphere. Meanwhile, 1-nitropyrene (1-NP), the most abundant nitro-polycyclic aromatic hydrocarbon found in particulate matter, is known to have carcinogenic effects; however, studies on its toxicity in human and canine skin are still needed. In this study, we investigated 1-NP-induced apoptosis and inflammatory pathways in HaCaT cells. In addition, we also measured the cytoprotective effect of methyl gallate (MG), which is widely distributed in medicinal and edible plants and is well known for its anti-inflammatory and antioxidant properties. MG inhibited 1-NP-induced cell death and apoptosis pathways, including the cleavage of PARP and activation of caspase-3, -7, and -9. MG also suppressed 1-NP-induced COX-2 expression and phosphorylation of mitogen-activated protein kinases (MAPKs) and MAPK kinases (MAPKKs). Our findings suggest that 1-NP induces skin toxicity in human and canine through apoptosis and inflammatory responses, and moreover, that this can be prevented by treatment with MG.

• Asian Journal of Atmospheric Environment
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• 제4권1호
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• pp.1-8
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• 2010

The effectiveness of the government regulation on tail-pipe emission for diesel vehicles issued in 2003 in Tokyo was evaluated in this study. Variations in annual average concentrations of polycyclic aromatic hydrocarbons (PAHs) and nitro-PAHs associated with airborne particulate matter were investigated in connection with the variation in airborne elemental carbon (EC) concentration in downtown Tokyo, Japan in 2006-2007 and in 1997-1998. The annual average concentrations of EC, seven different PAHs, and 1-nitropyrene were found to have decreased significantly from 1997-1998 to 2006-2007. The most prominent decrease in atmospheric concentration was observed for 1-nitropyrene, which is a representative nitro-PAH originating from diesel vehicles. This indicated that the government control has worked to considerably reduce both atmospheric mutagens and airborne particulate matter. In contrast, the concentrations of two nitro-PAHs, 2-nitrofluoranthene and 2-nitropyrene, remained the same. These nitro-PAHs are known to be formed by atmospheric nitration of their parent PAHs, and this result suggested factors other than the concentration of parent PAHs and $NO_2$ affects the degree of atmospheric formation of nitro-PAHs.

• Asian Journal of Atmospheric Environment
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• 제5권3호
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• pp.146-151
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• 2011

1-Nitropyrene (1-NP), 2-NP and 2-nitrofluoranthene (2-NFR) are useful markers for studying the atmospheric behaviors of polycyclic aromatic hydrocarbons (PAHs) and nitropolycyclic aromatic hydrocarbons (NPAHs). However, present methods for measuring trace levels of these compounds are lesssensitive and laborious. Here we describe several improvements to a previously reported high-performance liquid chromatography-chemiluminescence detection system that allows it to determine trace levels of 1-, 2-NPs and 2-NFR. The proposed system was equipped with a reducer column packed with Pt/Rh instead of zinc whose life-time was limited. The combination of Cosmosil MS-II (monomeric ODS) and AR-II (polymeric ODS) columns was used instead of polymeric ODS columns as the separator column to improve the separation. An ethanol mixture with acetate buffer (pH 5.5) was used in place of an acetonitrile mixture with the same buffer to activate the reducer column. The same ethanol mixture was used as the mobile phase for the clean-up column. The switching time of the column switching valve was optimized to concentrate the amino-derivatives of above NPAHs quantitatively on the concentrator column. The concentrations of bis(2,4,6-trichlorophenly) oxalate and hydrogen peroxide in the chemiluminescence reagent solution were optimized to 0.4 mM and 30 mM, respectively, to increase the sensitivity. Under the above conditions, the detection limits (S/N=3) of 1-, 2-NPs and 2-NFR were 1 fmol (0.25 pg), 10 fmol (2.5 pg) and 4 fmol (1 pg), respectively. The proposed system was effectively used to determine trace levels of 1-, 2-NPs and 2-NFR in airborne particulates collected at Noto Peninsula. The atmospheric concentrations of 1-, 2-NPs and 2-NFR were not more than sub pg $m^{-3}$ levels. They were higher in winter (January) than in summer (July). In both seasons, the concentrations were in decreasing order, [2-NFR]>[1-NP]>[2-NP].

• KSBB Journal
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• 제18권3호
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• pp.217-221
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• 2003

Ames test (S. typhimurium TA98, TA100)에서 6종의 직접변이원에 대한 가시오갈피(뿌리, 줄기, 잎)의 메탄을 추출물 및 용매 분획물의 항돌연변이 효과를 고찰하였다. Salmonella typhimuriun TA98, TA100에서, 6종의 직접변이원으로 유도된 각각의 변이원성에 대한 가시오갈피 뿌리, 줄기 및 잎의 메탄을 추출물의 돌연변이 억제효과를 검색한 결과 1-nitropyrene의 변이원성에 대하여 선택적으로 강한 억제효과를 나타내었다. 따라서 1-nitropyrene 변이원의 돌연변이원성에 대한 용매분획물의 활성을 비교한 결과 클로로포름 분획물이 가장 강한 억제효과를 나타내었다. S. typhimurium TA98, TA100에서, 뿌리의 클로로포름 분획물은 세포내 항돌연변이 효과를 강하게 나타낸 반면 줄기와 잎은 세포내 돌연변이 억제효과를 나타내지 않았다. 이 결과로부터 가시오갈피 뿌리에는 주로 세포내 항돌연변이 효과를, 줄기와 잎에는 세포외 항돌연변이 효과를 강하게 나타내는 활성 성분이 함유되어 있음을 알 수 있었다.

• Asian Journal of Atmospheric Environment
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• 제10권2호
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• pp.90-98
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• 2016

Total suspended particulates (TSP) in the atmosphere were collected for 2 weeks during winter in Muroran, Hokkaido, Japan, a typical iron-manufacturing city. The concentrations of polycyclic aromatic hydrocarbons (PAHs) and nitropolycyclic aromatic hydrocarbons (NPAHs) in TSP were determined by high-performance liquid chromatography (HPLC) using fluorescence and chemiluminescence detectors, respectively. No relationship was observed between the atmospheric PAH and NPAH concentration, or the atmospheric PAH and TSP concentration. However, there was a tendency that the atmospheric PAH concentration was higher when the wind blew from the coke-oven plant. Furthermore, the concentration ratios of 1-nitropyrene to pyrene, which is a suitable indicator of the contribution made by automobiles and coal combustion systems to urban air particulates, were smaller in Muroran and the values were close to those observed in particulates from coal combustion systems. Therefore, these results show that the PAH and NPAH compositions for Muroran are characteristic of an iron-manufacturing city.

• Environmental Analysis Health and Toxicology
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• 제7권3_4호
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• pp.69-79
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• 1992

Recently, concerns of water pollution and health risks caused by synthetic detergents have emerged, as the use of various detergents has increased It has been suggested that some surfactants are cocarcinogens. The surfactants tested were linear alkylbenzene sulfonate, sodium lauryl sulfate, polyoxyethylene sorbitan monooleat (tween 80), and the mutagens were 1-nitropyrene, N -methyl- N'-nitro-N -nitrosoguanidine, benzo (a) pyrene, and aflatoxin B$_1$. This study was undertaken to investigate the effects of surfactants on the activity of mytagens using the Ames mutagenic assay with Salmonella typhimurium TA98, TA100. The results were summarized as follows: 1. The surfactants have no mutagenic activity of themselves. 2. Higher doses of surfactants than 100 $\mu\textrm{g}$/plate reduced the number of revertants. It is assumed that the reduction would inhibited cell growth. 3. When the comutagenic ratio is defined as the ratio between mutagenic activity itself and the activity with mutagen and surfactant (drinking water quality standard), LAS showed the comutagenic ratio 0.86-1.17 and SLS 0.74-1.10 as well. According to the comparisons, it could not be recognised for the comutagenicity of drinking water quality standard of surfactant. 4. As increasing the amount of mutagens, the designated amount of surfactant did not affected the mutagen's activity statistically. From the above result, syunthetic surfactants do not present mutagenicity and comutagenicity in the microbial assay.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2001년도 추계학술대회
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• pp.157-163
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• 2001

Mycolic acid-containing gram-positive bacteria, so called nocardioform actinomycetes, have become a great interest to environmental microbiologists due to their metabolic versatility, multidegradative capacity and potential for bioremediation of priority pollutants. For example, Rhodococcus rhodochrous N75 was able to metabolize 4-methy1catechol via a modified $\beta$-ketoadipate pathway whereby 4-methylmuconolactone methyl isomerase catalyzes the conversion of 4-methylmuconolactone to 3-methylmuconolactone in order to circumvent the accumulation of the 'dead-end' metabolite, 4-methylmuconolactone. R. rhodochrous N75 has also shown the ability to transform a range of alkyl-substituted catechols to the corresponding muconolactones. A novel 3-methylmuconolactone-CoAsynthetase was found to be involved in the degradation of 3-methylmuconolactone, which is not mediated in a manner analogous to the classical $\beta$-ketoadipate pathway but activated by the addition of CoA prior to hydrolysis of lactone ring, suggesting that the degradative pathway for methylaromatic compounds by gram-positive bacteria diverges from that of proteobacteria. Mycobacterium sp. Strain PYR-l isolated from oil-contaminated soil was capable of mineralizing various polyaromatic hydrocarbons (PAHs), such as naphthalene, phenanthrene, pyrene, fluoranthrene, 1-nitropyrene, and 6-nitrochrysene. The pathways for degradation of PAHs by this organism have been elucidated through the isolation and characterization of chemical intermediates. 2-D gel electrophoresis of PAH-induced proteins enabled the cloning of the dioxygenase system containing a dehydrogenase, the dioxygenase small ($\beta$)-subunit, and the dioxygenase large ($\alpha$)-subunit. Phylogenetic analysis showed that the large a subunit did not cluster with most of the known sequences except for three newly described a subunits of dioxygenases from Rhodococcus spp. and Nocardioides spp. 2-D gel analysis also showed that catalase-peroxidase, which was induced with pyrene, plays a role in the PAH metabolism. The survival and performance of these bacteria raised the possibility that they can be excellent candidates for bioremediation purposes.