• Proceedings of the Korean Society of Toxicology Conference
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• 2000.11a
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• pp.45-67
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• 2000

2-Amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) is a mutagenic and carcinogenic heterocyclic amino found in cooked meat. The in vivo mutagenicity and hepatocarcinogenicity of MeIQx were examined in mice harboring the lacZ mutation reporter gene ($Muta^{TM}$ Mice) and bitransgenic mice over-expressing the c-myc oncogene. C57B1/$\lambda$lacZ and bitransgenic c-myc (albumin promoter)/$\lambda$lacZ mice were bred and weaned onto an AIN-76 based diet containing $0.06\%$ (w/w) MeIQx or onto control diet. After 30 weeks on diet, only male bitransgenic mice on MeIQx developed hepatocellular carcinoma ($100\%$ incidence) indicating that there was synergism between c-myc over-expression and MeIQx. By 40 weeks, hepatic tumor incidence was $100\%$ ($17\%$) and $44\%$ ($0\%$) in male c-myc/$\lambda$lacZ and C57B1/$\lambda$lacZ mice given MeIQx (or control) diet, respectively, indicating that either MeIQx or c-myc over-expression alone eventually induced hepatic tumors. At either time point, mutant frequency in the lacZ gene was at least 40-fold higher in MeIQx-treated mice than in control mice of either strain. These findings suggest that MeIQx-induced hepatocarcinogenesis is associated with MeIQx-induced mutations. Elevated mutant frequency in MeIQx-treated mice also occurred concomitant with the formation of MeIQx-guanine adducts as detected by the $^{32}P$-postlabeling assay. Irrespective of strain or diet, sequence analysis of the lacZ mutants from male mouse liver showed that the principal sequence alteration was a single guanine-base substitution. Adenine mutations, however, were detected only in animals on control diet. MeIQx-fed mice harboring the c-myc oncogene showed a l.4-2.6-fold higher mutant frequency in the lacZ gene than mice not carrying the transgene. Although there was a trend toward higher adduct levels in c-myc mice, MeIQx-DNA adduct levels were not significantly different between c-myc/$\lambda$lacZ and C57B1/$\lambda$lacZ mice after 30 weeks on diet. Thus, it appeared that factors in addition to MeIQx-DNA adduct levels, such as the enhance rate of proliferation associated with c-myc over-expression, may have accounted for a higher mutant frequency in c-myc mice. In the control diet groups, the lacZ mutant frequency was significantly higher in c-myc/$\lambda$lacZ mice than in 057B1/$\lambda$1acZ mice. The findings are consistent with the notion that c-myc over-expression is associated with an increase in mutagenesis. The mechanism for the synergistic effects of c-myc over-expression on MeIQx hepatocarcinogenicity appears to involve an enhancement of MeIQx-induced mutations.

• BMB Reports
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• v.34 no.1
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• pp.90-94
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• 2001

The mutagenic activity of chitosan oligosaccharide and its antimutagenic effect against 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) were investigated using the Salmonella/Ames test. No mutagenic activity was found in the Salmonella typhimurium strains TA 98 and TA 100, either with or without S9 activation. In contrast, chitosan oligosaccharide showed an inhibitory effect on the mutagenic activity of the cooked food mutagen, MeIQx, in the presence of S9. The influence of chitosan oligosaccharide on the genotoxicity of MeIQx was examined using a host-mediated assay in mice. The oligosaccharide was administered for 14 consecutive days (intragastric application at doses of 0.1 or 0.5 g/kg body wt) to mice. S. typhimurium TA 98 was given intravenously before an oral dose of MeIQx (4.5 mg/kg body wt.). The number of $his^+$ revertants were determined from the Ever of mice. The intragastric application of oligosaccharide led to a 47% reduction in the number of mutants induced by MeIQx (p<0.05). These results suggested that chitosan oligosaccharide had antimutagenic properties against MeIQx in vitro and in vivo.

• Journal of Microbiology and Biotechnology
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• v.11 no.2
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• pp.346-349
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• 2001

The antimutagenic activity of polysaccharides extracted from soybeans fermented with Agrocybe cylindracea (AC) or Phellinus igniarius (PI) against 2-amino-3,8-dimethylimidaxo[4,5-f]quinoxaline (MeIQx) was examined using a Salmonella/Ames test and host-mediated assay in mice. The polysaccharides from the soybeans fermented with A. Cylindracea and P. igniarius inhibited the mutagenic acitivity of the cooked food mutagen, MeIQx, by 31.2% and 35.3%, respectively. The polysaccharides also inhibited MeIQx genotoxicity in a dose-dependent manner in micel. These results suggest that the polysaccharides from soybeans fermented with A. cylindracea or P. igniarius exhibit antimutagenic properties against MeIQx in vitro and in vivo.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.3
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• pp.537-542
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• 1998

The present study was performed to elucidate desmutagenic principles from Ecklonia stolonifera extracts against 2-amino-1-methyl-6-phenylimidazo[4,5-b] pyridine(PhIP) and 2-amino-3,8-dime-thylimidazo[4,5-f]duinoxaline(MeIQx) with Salmonella/mammalian-microsome mutagenicity test. Alginate, phenols, chlorophyll and carotenoids from Ecklonia stolonifera were extracted and their desmutagenicities were assayed. Alginate hydroysates showed desmutagenic activities against PhIP and MeIQx at high level dose. Phenol fractions and bromophenol showed desmutagenic activity of about MeIQx at high level dose. Phenol fractions and bromophenol showed desmutagenic activity of about 90% per 0.5mg against PhIP and MeIQx. Chlorophyllin among chlorophyll derivatives exhibited remarkable desmutagenic activities of 92.9% and 82.7% at 20uM against PhIP and MeIQx, respectively. Carotenoids, such as lutein and $\alpha$-cryptoxanthin isolated from Ecklonia stolonifera exerted also high desmutagenic activity. Major desmutagenic substances from Ecklonia stolonifera are considered to be chlorophyllin, phenols, lutein, $\alpha$-cryptoxanthin and low molecular alginates.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.31 no.4
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• pp.581-586
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• 1998

The present study was conducted to prepare seaweed extracts suppressing mutagenic activity of 2-amino-1-methyl-6-phenylimidazo[4,5-b] pyridine ( PhIP ) ana 2-amino-3,8-dimethylmidazo[4,5-f]quinoxaline (MeIQx) derived from cooked meat produce. The tumor initiation activity of PhIP and MeIQx was assayed with Ames method using Salmonella typhimurium TA98 in the presence of S-9 mixtures before and after addition of methanol-solubles of seaweed, such as, Phaeophyta; Undaria pinnatifida, Ecklonia stolonifera Ecklonia cava, Laminaia japonica Sargassum fulvellum, Sargassum hornezi, Sargassum miyabei, Sargassum thunbergii, Agarum cribrosum and Hizikia fusifomis, Rhodophpei Porphyra yezoensis, Grateloupia eiliptiut Lomentraria catenata, Ploemium telfairiae and Glarilaria verrucosa, Chlorophyta; Codium fragile, Enteromorpha compresa and Ulva pertusa. Among seaweed tested, Phaeophyta was shown the higher desmutagenic activity than Rhodophyta and Chlorophyta. E. stolonifera, E. cava and S. miyabei, among Phaeophyta exerted the stronger desmutagenic activity (above $90\%$/2 mg). The ethyl acetate, diethyl ether and chlomform extracts except water extracts from E. stolonifera exhibited a high desmutagenic activity. The ethyl acetate extract of E. stolonifera which showed highest activity was fractionated with Sephadex LH20 column chromatography to give active fraction A-7, which showed desmutagenicity of $90\%$/mg against PhIP and $80\%$/mg against MeIQx. The active fraction had the absorbance at 207.7 and 232nm.

• Food Science of Animal Resources
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• v.31 no.4
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• pp.563-569
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• 2011

Heterocyclic amines (HCAs) are food mutagens and carcinogens that are found in cooked fish, meat, and protein-rich foods. This study examined the inhibitory effect of marinades containing a Nelumbo nucifera leaf fraction on HCA formation in cooked beef steak. As a result of the Ames assay, cooked beef marinated with the N. nucifera leaf butanol fraction (2.0 g) cooked at $190^{\circ}C$ showed a 61.5% reduction in overall mutagenicity. However, these data revealed no significant difference in mutagenicity in the ethanol, ethyl acetate, or water fractions. Formation of MeIQx (2-amino-3,8 dimethylimidazo[4,5-f]-quinoxalin) and PhIP (2-amino-1-methyl-6-phenylimidazo[4,5-b]-pyridine) was inhibited 60.7-63.5%. Cooked beef marinated with the water fraction of the N. nucifera leaf significantly (p<0.05) reduced the formation of MeIQx, PhIP, and DiMeIQx (2-amino-3,4,8 trimethylimidazo[4,5-f]-quinoxaline) by 65.3, 67.6, and 73.9%, respectively. These results show that marinades containing the N. nucifera leaf fraction could be an alternative method for reducing HCA formation in cooked beef steak.

• Toxicological Research
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• v.34 no.4
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• pp.291-296
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• 2018

Chemical carcinogenesis is a multistep process. Genotoxic carcinogens, which are DNA-reactive, induce DNA adduct formation and genetic alterations in target cells, thereby generating mutated cells (initiation). Subsequently, preneoplastic lesions appear through clonal proliferation of the mutated cells and transform into tumors (promotion and progression). Many factors may influence these processes in a dose-dependent manner. Therefore, quantitative analysis plays an important role in studies on the carcinogenic threshold of genotoxic carcinogens. Herein, we present data on the relationship between key carcinogenic events and their deriving point of departure (PoD). Their PoDs were also compared to those of the carcinogenesis pathway. In an experiment, the liver of rats exposed to 2-amino-3,8-dimethylimidazo-(4,5-f)quinoxaline (MeIQx) was examined to determine the formation of MeIQx-DNA adducts, generation of mutations at LacI transgene, and induction of preneoplastic glutathione S-transferase placental form (GST-P)-positive foci and tumors (benign and malignant). The PoDs of the above key events in the carcinogenicity of MeIQx were increased as the carcinogenesis advanced; however, these PoDs were lower than those of tumor induction. Thus, the order of key events during tumor induction in the liver was as follows: formation of DNA adducts ${\ll}$ Mutations ${\ll}$ GST-positive foci (preneoplasia) ${\ll}$ Tumor (adenoma and carcinoma). We also obtained similar data on the genotoxic and carcinogenic PoDs of other hepatocarcinogens, such as 2-amino-3,8-dimethylimidazo(4,5-f)quinoline. These results contribute to elucidating the existence of a genotoxic and carcinogenic threshold.

• Environmental Mutagens and Carcinogens
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• v.18 no.1
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• pp.26-31
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• 1998

Antimutagenic activity of Saururus chinesis (Lour.) Bail was investigated for food-borne mutagens using S. typhimurium TA98. Methanol extract from Saururus Chinesis (Lour.) Bail was fractionated into hexane, chloroform, ethylacetate and butanol fractions, followed by determination of antimutagenic activity for food-borne mutagenic heterogenic amines (HCA). The hexane fraction exhibited a strong antimutagenic activity for 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), 2-amino-3,8-dimethylimidazo[4,5-f] quinoxaline (MeIQ), 2-amino-3,4-dimethyl-3H-imidazo[4,5-f]quinoline (MeIQx), 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), 3-amino-1-methyl-5H-pyroid[4,3-b]indole acetate (Trp-2-A); however its fraction rather enhanced the bacterial mutagenicity of 2-amino-3,4,8-trimethyl-3H-imidazo[4,5-f]quinozaline (4,8-diMeIQx) and 2-amino-3,7,8-trimethyl-3H-imidazo[4,5-f]quinoxline (7,8-diMeIQx). Active principle in the fraction was found to be two major compounds (${\gamma}$-crene B and epi-bicyclosesquiphellandrane) and 6 minor compounds (${\delta}$-caryophyllene, ${\gamma}$-elemene, ${\beta}$-cabebene, ${\delta}$-cadinene, ${\delta}$-selinene, and patchoulene). Modulation effect for the mutagenic activity of the food-borne mutagenic HCA by the fraction might be derived from a cumulative effect of each individual compounds. Hence, this hexane fraction might be use to reduce the production of mutagenic HCA during cooking process of protein-rich foods.

• Korean Journal of Food Science and Technology
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• v.40 no.4
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• pp.389-393
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• 2008

This study examined the effects of cooking temperature and onion (Allium cepa L.) tissue concentrate on heterocyclic amine (HCA) formation in fried ground beef patties. Various amounts of onion tissue (2.0, 5.0, and 10.0%, w/w) were added to the ground beef patties, which were then fried at two different temperatures (190 and $225^{\circ}C$) for 10 min/side. The ground beef patties fried at $190^{\circ}C$ and containing 10.0% (w/w) onion showed a 51% decrease in mutagenicity, and formation of 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]-pyridine (PhIP) was reduced by 58% and 63%, respectively. For the patties fried at $225^{\circ}C$, total mutagenicity decreased by 35% and 48% with the additions of 5.0 and 10.0% (w/w) onion, respectively, and PhIP formation was reduced 50, 60, and 71% with the additions of 2.0, 5.0, and 10.0% (w/w) onion, respectively.

• Food Science of Animal Resources
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• v.28 no.5
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• pp.637-644
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• 2008

Four different extraction and purification methods were evaluated to determine the heterocyclic amines (HCAs) in fried pork patties. Pork patties were cooked in the teflon-coated electric frying pan at $230^{\circ}C$ for 8 min per side. HCAs in the fried pork patties were extracted and purified using four different solid-phase extraction (SPE) methods and quantitated by LC-MS (API-ESI). Recovery of four different extraction and purification methods was evaluated by comparing the HCAs amounts quantified by the standard addition method. Validation of extraction and purification methods for fried pork patties was determined to establish accurate sample preparation. The recoveries of HCAs from different SPE methods were calculated. The recovery yields were 15.7-68.7% (Polar amine group) and 25.0-74.7% (less-polar amine group) in method A. Method D provided recovery yields ranging from 14.1% to 68.7% in polar amine groups and from 3.0% to 72.3% in less-polar amine groups, respectively. Modified procedures of Method A and D were the most suitable extraction and purification method for HCAs analysis from fried pork patties.