• Toxicological Research
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• 제18권2호
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• pp.129-138
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• 2002

The mouse lymphoma assay (MLA) has been recently validated as a sensitive and specific test system to determine the genotoxic potential for a chemical. The objective of this study is to evaluate the utility of MLA for detecting mutagens. Especially, to compare MLA with the in vitro chromosomal aberration test (CA), we performed MLA using the microwell method with three chemicals (hydroxyurea, theophylline and amino acid copper complex), which were reportedly positive in the CA. In cell treated with hydroxyurea, anti-neoplastic agent that blocks DNA replication, evidence of a positive response was obtained without S9 mix for 4 h and 24 h. In addition, analysis of colony size distribution at concentration that gave an elevated mutant fraction showed that hydroxyurea induced a high proportion of small type colonies, indicating that hydroxyurea-induced mutation is associated with large chromosomal deletion. Conversely, negative MLA result was obtained for theophylline, which was wed as central nervous system stimulator. Although theophylline increased the mutant frequency at concentration of 1250 $\mu\textrm{g}$/$\textrm{m}{\ell}$ with S9 mix for 4 h, a concentration-related increase in mutant frequency was not observed. The MLA result of amino acid copper complex was considered equivocal because the positive result was obtained at concentration showing 10% or less RS or RTG. Thus, among 3 CA-positive chemicals, positive MLA result was obtained for one. The other two chemicals were negative and equivocal. However MLA, which evaluates mutagenic potential of chemicals through colony formation by cell grouth, may provide a higher predictivity of carcinogenesis than CA.

• Journal of Technologic Dentistry
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• 제22권1호
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• pp.145-152
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• 2000

In order to select an effective antibiotic substance against oral resident bacteria, we were isolated from soil and texonomically analyzed. Seven hundred and eighteen strains were isolated on humic acid- vitamin agar(HV agar) and 220 strains were on methanol medium from three each paddy forest, field and riverside soil samples. So, during the screening of antibiotics from soil, we isolated microorganisms showing powerful antagonistic activity against oral resident bacteria. Microorganism was tested against 25 strains of bacteria, yeast and fungi. Among them, No. 248 strain exhibited the most strongly growth inhibition. So, the taxonomical analysis the isolated strain was found to be unknown Actinomyces sp. and was named No 248. A production of the antibiotics from No. 248 begins at the early exponential phase developed at the 72th hour under the optinum conditions. The property of No. 248 antimicrobial compound was very stable under acid(pH 3.0) and alkali(pH 10.0) treatment, but it was instable in heat treatment at $120^{\circ}C$. For the improvement of antibiotic activity, two mutants were isolated from strain No. 248 by the treatment of mutagenic agents, NTG and hydroxylamine. As a result, the mutant strains excreted the potent antibiotics to inhibit the growth of Candida albicans.

• Mycobiology
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• 제33권2호
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• pp.104-108
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• 2005

Genus Phellinus taxonomically belongs to Aphyllophorales and some species of this genus have been used as a medicinal ingredients and Indian folk medicines. Especially, P. linteus and morphological-related species are well-known medicinal fungi that have various biological activities such as humoral and cell-mediated, anti-mutagenic, and anti-cancer activities. However, little is known about the rapid detection for complex Phellinus species. Therefore, this study was carried out to develop specific primers for the rapid detection of P. linteus and other related species. Designing the species-specific primers was done based on internal transcribed spacer sequence data. Each primer set detected specifically P. linteus (PL2/PL5R) and P. baumii (PB1/PB4R). These primer sets could be useful for the rapid detection of specific-species among unidentified Phellinus species. Moreover, restriction fragment length polymorphism analysis of the ITS region with HaeIII was also useful for clarifying the relationship between each 5 Phellinus species.

• Journal of Environmental Science International
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• 제2권1호
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• pp.57-68
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• 1993

Polycyclic aromatic hydrocarbons (PAMs) are ubiquitous contaminants in marine environments. PAHs enter estuarine and nearshore marine environment via several routes such as combustion of fossil fuels, domestic and industrial effluents and oil spills PAHs have been the focus of numerous studies in the world because they owe potentially carcinogenic, mutagenic, and teratogenic to aquatic organisms and humans from consuming contaminated food. However, one can hardly find any available data on PAM content in marine organisms in Korea. The present study was carried out in order to determine PAH content in oysters from the intertidal and subtidal zones of Chinhae Bay, which is located in near urban communities and an industrial complex, and the bay is considered to be a major repositories of PAHs. 16 PAHs were analyzed by High Performance Liquid Chromatography (HPLC) with uv/vis and fluorescence detectors in oysters: they are naphthalene (NPTHL), acenaphthylene (ANCPL), acenaphthene (ACNPN), fluorene (FLURN), phenanthrene (PKEN), anthracene (ANTHR), fluoranthene (FLRTH), pyrene (PYRf), benzo(a)anthracene (BaA), chrysene (CHRY), benzo(b)- fluoranthene (BbF), benzo(k)fluoranthene (BkF), benzo(a)pyrene (BaP), dibenz(a, h)anthracene (DhA), benzo(g, h, i)peryne (Bghip) and indeno(1, 2, 3, -cd)pyrene (I123cdP). The PAH contents in oysters from the intertidal and subtidal zones of Chinhae Bay ranged from < 0.1 to 992.0 $\mu\textrm{g}$/kg (mean 69.8 $\pm$ 9.8 $\mu\textrm{g}$/kg). Key words . polycyclic aromatic hydrocarbon, high performance liquid chromatography, oyster, Chinhae Bay.

• Horticultural Science & Technology
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• 제29권1호
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• pp.53-60
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• 2011

Cytoplasmic male sterility (CMS) induced by mutant mitochondria genome, has been used for commercial seed production of $F_1$ hybrid cultivars in diverse crops. In pepper (Capsicum annuum L.), two sterile cytoplasm specific gene organization, atp6-2 and coxII were identified. An open reading frame, orf456 nearby coxII gene has been speculated to induce male sterility (MS) by mutagenic analysis. Moreover, molecular markers for atp6-2 and coxII of mitochondrial genotype (mitotype) were developed. However, the Cytoplasmic MS specific markers, atp6SCAR and coxIISCAR markers appeared in both N and S cytoplasms when polymerase chain reaction (PCR) cycles prolonged more than 40 cycles. Since the reported molecular markers were dominant markers, the presence of the faint sterile-specific band in normal cytoplasm may lead to the mis-classification of pepper breeding lines. To solve this problem, one common forward primer and two different reverse primers specific to normal coxII and sterile orf456 genes were designed after analyzing their gene organizations. By using these three primers, N and S coxII specific bands were co-amplified in male-sterile lines, but only normal coxII specific band was amplified in maintainer lines. Since the reverse primer for sterile coxII was specifically designed 275 bp downstream of orf456, relatively stable PCR amplification patterns were observed regardless of the number of PCR cycles. These primer sets easily identified different mitotypes among the divergent breeding lines, commercial cultivars and diverse germplasms.

• Archives of Pharmacal Research
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• 제18권6호
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• pp.415-422
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• 1995

Twenty-four flavones were synthesized with various hydroxyl and/or methoxyl groups on A and B rings. Their antimutagenic properties were evaluated against ben:w(a)pyrene (BaP) and a pool of mutagenic urine concentrate (U) using a modified liquid incubation method of Ames test. The tester strain was Salmonella typhimurium TA98+S9 Mix. The antimutagenic activities were calculated by non linear regression analysis and the doses of flavones (in nmoles) required for a 50% reduction of induced revertants with BaP and U were defined as the inhibition doses (TEX>$ID_{508}{\;}and{\;}ID_{508}$ respectively). Seventeen flavones possessed significant antimutagenic activity against BaP. $ID_{508}$ ranged from 15.1 nmoles (F22) to 1000.6 nmoles (F13). Eighteen f1avones showed significant antimutagenic activity against U. $ID_{50U}$ ranged from 23.5 nmoles (F22) to 354.6 nmoles (F3). The 2',3',4'-trihydroxyflavone (F22, $ID_{508}=15.1$ nmoles, $ID_{50U}=23.5$ nmoles) and the 2',3',4',7-tetrahydroxyflavone (F20, $ID_{508}=37.8$ nmoles; $ID_{50U}=62.3$ nmoles) had antimutagenic activities similar to those of chlorophyllin ($ID_{508}=19.6$ nmoles and $ID_{50U}=44.2$ nmoles) and were evaluated against B(alP 7,8-dihydrodiol-9,10-epoxide. Against this last mutagen, the flavones which included three OH in B ring showed the highest activity and this property seemed independent of the substituent groups on A ring.

• Toxicological Research
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• 제31권3호
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• pp.255-264
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• 2015

Heterocyclic amines (HCAs) and acrylamide are unintended hazardous substances generated by heating or processing of foods and are known as carcinogenic and mutagenic agents by the animal experiments. A simple method was established for a rapid and accurate determination of 12 types of HCAs (IQ, MeIQ, Glu-P-1, Glu-P-2, MeIQx, Trp-P-1, Trp-P-2, PhIP, $A{\alpha}C$, $MeA{\alpha}C$, Harman and Norharman) and acrylamide in three food matrices (non-fat liquid, non-fat solid and fat solid) by isotope dilution liquid chromatography-tandem mass spectrometry (LC-MS/MS). In every sample, a mixture of internal standards including $IQ-d_3$, $MeIQx-d_3$, $PhIP-d_3$, $Trp-P-2-^{13}C_2-^{15}N$ and $MeA{\alpha}C-d_3$ was spiked for quantification of HCAs and $^{13}C_3$-acrylamide was also spiked for the analysis of acrylamide. HCAs and acrylamide in sample were extracted with acetonitrile and water, respectively, and then two solid-phase extraction cartridges, ChemElut: HLB for HCAs and Accucat: HLB for acrylamide, were used for efficiently removing interferences such as pigment, lipid, polar, nonpolar and ionic compounds. Established method was validated in terms of recovery, accuracy, precision, limit of detection, limit of quantitation, and linearity. This method showed good precision (RSD < 20%), accuracy (71.8~119.1%) and recovery (66.0~118.9%). The detection limits were < 3.1 ng/g for all analytes. The correlation coefficients for all the HCAs and acrylamide were > 0.995, showing excellent linearity. These methods for the detection of HCAs and acrylamide by LC-MS/MS were applied to real samples and were successfully used for quantitative monitoring in the total diet study and this can be applied to risk assessment in various food matrices.

• 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.

• Environmental Analysis Health and Toxicology
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• 제5권3_4호
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• pp.19-28
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• 1990

Nitroarenes are ubiquitous environmental pollutants displaying potent mutagenicity in bacteria and carcinogenicity in mammal. In this study, the concentration of nitroarenes in coarse and fine particles and mutagenicity of POC$\_$N/ fraction was investigated in suspended particulates at the Shinchon and Bulkwang area of Seoul. The suspended particulates were collected bimonthly by a high volume cascade impactor air sampler from July 1987 to May 1988. Extractable organic matter was obtained by ultrasonic extraction on diethly ether/cyclohexane (8/2, v/v). Neutral fraction was obtained by liquid-liquid extraction. Polar neutral organic compounds (POC$\_$N/) was fractionated by thin-layer chromatography. Finally, the concentrations of nitroarenes in POC$\_$N/ fraction were measured and determined by capillary gas chromatography. Direct and indirect mutagenicity of POC$\_$N/ fraction were measured using Salmonella typhimurium TA 98. The result were as follows: 1) Major nitroarenes at the Shinchon area was 1-nitropyrene and at the Bulkwang area it was 2,7-dinitro-9-fluorenone during the year. 2) Average concentration of total nitroarenes measured was 67.26 ng/m$^3$in fine particles which was 1,3 folds higher that in coarse particle (52.30 ng/m$^3$). 3) Annual pattern of nitroarenes concentrations revealed that concentration during heating season (Feb., Jan., Mar.) was 2.2 folds higher than that in non heating season (May, Jul., Sep.). Concentration of each season has 157.68 ng/m$^3$and 80.39 ng/m$^3$. 4) The mutagenic activity of POC$\_$N/ fraction from fine particles was higher compared to that of coarse particles and was increased when metabolically activated, with 59 mixture. Mutagenicities, Metabolically activated, were significantly different between Shinchon and Bulkwang area, 322.8 rev/250 $\mu\textrm{g}$/plate and 286.8 rev/250 $\mu\textrm{g}$/plate, respectively. 5) Annual pattern of mutagenicity of POC$\_$N/ fraction revealed that mutagenicity during the heating season was 1.7 folds higher at Shinchon area and 1.2 folds higher at Bulkwang than during the non heating season. The variable contents and levels of nitroarenes in suspended particulates may affect human health significantly. Further studies such as risk assessment should be conducted on the basis of these kind of studies.

• Analytical Science and Technology
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• 제17권3호
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• pp.263-270
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• 2004

A method is described for the analysis of short-chain alkylphenol ethoxylates (APEOs), 4-octylphenol-di-ethoxylate (OP2EO) and 4-nonylphenol-di-ethoxylate (NP2EO), in drinking water or wastewater using reversed phase high-performance liquid chromatography with electrospray ionization mass spectrometry. The solvent system was water and methanol containing $10{\mu}M$ trifluoroacetic acid as an ionization solvent. We acidified 1 L of water samples to less than pH 2 with concentrated $H_2SO_4$ and loaded onto Sep-Pak $C_{18}$, and eluted with acetone. The calibration of OP2EO and NP2EO was performed for the concentration range from 20 to 500 ng/L and the correlation coefficients were 0.999 and 0.990, respectively. The limits of detection were 20 ng/L (OP2EO) and 50 ng/L (NP2EO) at a signal-to-noise ratio of 3. Accuracy and precision of this analytical method were 85.8 ~ 122.1% and 8.2 ~ 18.8%, respectively. The proposed method allowed a sensitive and rapid detection of OP2EO and NP2EO and it could be applied for monitoring of APEOs from environmental samples.