• 한국식품영양과학회지
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• 제28권3호
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• pp.599-606
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• 1999

Water extracts of persimmon leaf tea(PLTE), green tea(GTE) and oolong tea(OTE), at the con centration used for human consumption, were examined for inhibitory effects on the mutagenicity of major classes of dietary and environmental mutagens including indirect acting mutagens, B[ ]P (benzo[ ]pyrene), IQ(2 amino 3 methylimidazo[4,5 f]quinoline), 2 AA(2 aminoanthracene) in the presence of S9 mix and direct acting mutagen, 4 NQO(4 nitroquinoline 1 oxide) without S9 mix, using the modified Ames Salmonella/microsome assay. PLTE, GTE and OTE showed very potent and concentration dependent antimutagenic effects against indirect acting mutagens B[ ]P and IQ. At the maximum concentration(16,200 g/plate) of each tea extract, number of colonies decreased in a dose dependent manner up to 82~100%. Similar inhibition of PLTE, GTE and OTE were seen at higher concentration in the mutagenicity of the 2 AA following an initial increase in the activity at lower concentration. However, the mutagenicity of the direct acting mutagen 4 NQO were not suppressed at lower concentration of the three tea extracts, and higher concentration of the tea extracts enhanced mutagenic activity of the mutagen. There were no differences in the mode of antimutagenesis between PLTE, GTE, and OTE, in both Salmonella typhimurium TA98 and TA100 strains against the same mutagen. In conclusion, the water extracts of persimmon leaf tea, green tea and oolong tea possess marked antimutagenic potential against a variety of important dietary and environmental indirect acting mutagens, but the activity was not observed against the direct acting mutagens. These results suggest that the mode of inhibitory action may not have resulted from direct interaction between tea extracts and the mutagens, but rather from indirect metabolic inactivation of mutagens by tea extracts.

• Journal of Microbiology and Biotechnology
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• 제14권2호
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• pp.268-275
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• 2004

To investigate the potential application of the single-cell gel electrophoresis (SCGE) assay to carp as an aquatic pollution monitoring technique, gill, liver, and blood cells were isolated from carp exposed to a direct-acting mutagen, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), or indirect mutagen, $benzo[\alpha]pyrene$ $(B[\alpha]P)$, then the DNA strand breakage was analyzed using the assay. Based on testing 5 different cell isolation methods and 6 electrophoretic conditions, the optimized assay conditions were found to be cell isolation by filter pressing and electrophoresis at a lower voltage and longer running time (at 0.4 V/cm for 40 min). In preliminary experiments, gill and liver cells isolated from carp exposed to MNNG in vitro exhibited DNA damage signals even with 0.5 ppb exposure, which is a much higher dose than previously reported. In the gill cells isolated from carp exposed to 0.01-0.5 ppm MNNG in vivo, significant dose-and time-dependent increases were observed in the tail for 4 days. As such, the linear correlation between the relative damage index (RDI) values and time for each dose based on the initial 48-h exposure appeared to provide effective criteria for the genotoxicity monitoring of direct-acting mutagenic pollution. In contrast, the in vivo exposure of carp to 0.25-1.0 ppm of $B[\alpha]P$ for 7 days resulted in dose-and time-dependent responses in the liver cells, in which 24-h delayed responses for metabolizing activation and gradual repair after 48 h were also observed. Thus, the negative-sloped linear correlation between the RDI and time at each dose based on the initial 48 h appeared to provide more effective criteria for the genotoxicity monitoring of indirect mutagenic pollution.

• 한국식품영양과학회지
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• 제28권2호
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• pp.452-457
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• 1999

The inhibitory effect of Coprinus comatus on the mutagenicity in Salmonella assay system and SOS chromotest were studied. In Ames test, the ethanol and water extracts and the cultured mycelia fractions of Coprinus comatus did not show any mutagenicity, but the Coprinus comatus ethanol extracts showed inhibitory effects of 8 0∼90% on the mutagenicity induced by indirect mutagen of benzo(a)pyrene(B(a)P) and aflatoxin B1(AFB1) in Salmonella typhimurium TA98 and TA100. The antimutagenic effect increased with increasing concentration of the ethanol extract toward N methyl N' nitro N nitrosoguanidine(MNNG). However, the water extracts inhibited about 40∼50% against direct and indirect mutagen. The cultured mycelial filtrate of Coprinus comatus, the fractionⅡ, showed antimutagenic effect of 90% on MNNG and 25∼50% on B(a)P and AFB1. In SOS chromotest, the ethanol extracts of Coprinus comatus showed antimutagenic effect of 65∼81% on SOS function induced by 4 NQO, and the cultured mycelia fractionⅡ showed low inhibitory effect of 20∼50%.

• 생명과학회지
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• 제5권3호
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• pp.121-125
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• 1995

To determine whether the omega 3 family, linolenic acis(LnA) is effective to inhibit carcinogens/mutagens-induced mutagenesis, we employed the Ames test using Salmonella typhimurium strain of TA100 and the SOS chromotest using Escherichia coli PQ37 strain. The inhibitory effect of LnA shown in the Ames assaying system was 95%, 78% and 73% when the mutagenicities were mediated by AFB$_{1}$, MNNG and 4-NQO, respectively. LnA shows a strong antimutagenic activity against indirect mutagen of AFB$_{1}$, whereas the same concentration of LnA exhibited weaker inhibitory effects on the direct mutagen of MNNG and 4-NQO than that of AFB$_{1}$. However. LnA reduced more than 80% of SOS responses induced by MNNG and 4-NQO when the adding concentration increased to 5%. We conclude that LnA contains in vitro antimutagenic properties and that this finding warrants further investigation both in vitro and in vivo to assess its possible chemotherapeutic potential.

• Toxicological Research
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• 제23권3호
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• pp.207-214
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• 2007

Chronic oxidative stress produced by exposure to environmental chemicals or pathophysiological states can lead animals to aging, carcinogenesis and degenerative diseases. Indirect antioxidative mechanisms, in which natural or synthetic agents are used to coordinately induce the expression of cellular antioxidant capacity, have been shown to protect cells and organisms from oxidative damages. Electrophile and free radical detoxifying enzymes, which were originally identified as the products of genes induced by cancer chemopreventive agents, are members of this protective system. The NFE2 family transcription factor Nrf2 was found to govern expression of these detoxifying enzymes, and screening for Nrf2-regulated genes has identified many gene categories involved in maintaining cellular redox potential and protection from oxidative damage as Nrf2 downstream genes. Further, studies using Nrf2-deficient mice revealed that these mutant mice showed more susceptible phenotypes towards exposure to environmental chemicals/carcinogens and in oxidative stress related disease models. With the finding that cancer chemopreventive efficacy of indirect antioxidants (enzyme inducers) is lost in the absence of Nrf2, a central role of Nrf2 in the antioxidative protective system has been firmly established. Promising results from cancer prevention clinical trials using enzyme inducers propose that pharmacological interventions that modulate Nrf2 can be an effective strategy to protect tissues from oxidative damage.

• Toxicological Research
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• 제20권3호
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• pp.225-232
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• 2004

The suitability of the single cell gel electrophoresis (SCGE) assay as a test for the monitoring of genotoxicity of aquatic environment was evaluated. The SCGE assay was employed to detect DNA damage induced in clam (Spisula sachalinensis) exposed to a direct mutagen, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) or an indirect mutagen, benzo[a]pyrene (B[a]P). The cells of gill and digestive glands were isolated from clam by homogenization, which was the optimized cell dissociation method, and the level of DNA damage was assessed and expressed as mean tail length. In the gill cells, significant dose- and time-dependent increase was observed in the mean tail length at the concentration from 0.01 to 0.5 ppm MNNG for 96 h. The linear correlation between relative dam-age index (RDI) values was suggested to provide criteria of genotoxicity monitoring for direct acting mutagen. The dose- and time-dependent responses of the digestive glands cells were less sensitive than those of the gill cells. In contrast, the genotoxic response resulting from the exposure of 0.01～1.0 ppm B[a]P to clam revealed a higher sensitivity in the digestive glands cells than the gill cells. The comparison between the time profiles of genotoxic responses in clam and carp, the latter had been obtained in our previous study, indicated that the metabolism of genotoxic compounds in the two aquatic organisms were quite different each other. We conclude that the SCGE assay has the potential as a screening test for routine genotoxicity monitoring of aquatic organisms because of its higher sensitivity and simplicity.

• Preventive Nutrition and Food Science
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• 제2권4호
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• pp.348-353
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• 1997

The antimutagenic and cancer cell growth inhibitory effects of methanol extracts from 9 kinds of seaweed were studied in the Ames assay and cell culture systems, respectively. The methanol extracts from the seaweeds of sea lettuce, chlorella, sea tangle, sea mustard, sporophyll of sea mustard, fusiforme, seaweed papulosa, purple laver and ceylon moss showed antimutagenicities against aflatoxin B₁(AFB₁) and N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) in the Salmonella typhimurium TA100. These extracts revealed relatively higher antimutagenicity against AFB₁(indirect mutagen) than MNNG(direct mutagen). Sporophyll of sea mustard and seaweed papulosa exhibited strong antimutagenic activity against AFB₁, and sporophyll of sea mustard, sea tangle and ceylon moss also reduced the mutagenicity induced by MNNG. The sporophyll fo sea mustard exerted the highest antimutagenic activity among the samples treated. The methanol extracts from 9 kinds of seaweed inhibited the growth of two cancer cell lines, AGS human gastric adenocarcinoma cells and HT-29 human colon carcinoma cells. Sea tangle, sea mustard and sporophyll of sea mustard inhibited the growth of cancer cells significantly. These results suggest that various seaweeds show not only antimutagenic activity but also growth inhibitory effect of some cancer cells.

• 한국식품영양과학회지
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• 제24권3호
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• pp.440-445
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• 1995

The antimutagenic effects of juices from green pepper(GP), red pepper leaf(RPL), red pepper(RP) and sweet pepper(SP) were examined by the Ames method using Salmonella typhimurium TA100. The juice supernatants of GP, RPL and RP showed antimutagenic activities against afltoxin B1(AFB1) in Salmonella typhimurium TA 100. The juice supermatants of GP and RPL also exhibited the inhibitory effects(p<0.05) to the mutagenicities induced by N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) and 4-nitro-quinoline-1-oxide(4-NQO). The juice of RP showed antimutagenic activities against indirect mutagen of AFB1, however, the activity was reduced at higher concentration(5.0%), furthermore, as the adding concentration of sample increased to 5.0%, it exhibited slight comutagenicith on direct mutagen of MNNG. Theantimutagenic activities of GP and RPL juices were reduced significantly after heating at 100℃ for 20min, supposing that the antimutagenic compound(s) in the juices were heat labile.

• 한국식품과학회지
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• 제36권2호
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• pp.317-320
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• 2004

Curcumin의 추출효율은 에탄을 추출방법이 가장 높았고, 항산화효과는 에탄올, 초임계, 열수 추출물 순으로 나타났다. Ames의 mutagenicity test를 실시한 결과 각각의 추출물은 돌연변이원이 없는 것으로 확인되었고, TA98에 직접변이원 2-NF 처리시 초임계 추출물의 저해율이 20.1%로 에탄올과 열수 추출물에 비하여 가장 높은 항돌연변이 효과를 나타냈다. 또한 간접 변이원 2-acetamidofluorene($10\;{\mu}L/plate$; Sigma Co.; 2-AF)에 대한 항돌연변이 효과를 알아본 결과 2-AF로 유도된 TA98은 초임계 추출물을 0.1 mL/plate로 처리시 항돌연변이의 효과는 12.2%였으며, 알코올 추출물과 열수 추출물의 경우는 항돌연변이 효과가 나타나지 않았다. Nitrite의 분해능의 경우에 있어서도 에탄올 및 열수 추출물에 비하여 초임계 추출물이 높은 분해율을 나타냈으며 pH 1.2에서 가장 높은 분해율을 보였다. 이와 같이 초임계를 이용해 추출한 curcumin은 항산화효과, nitrite 분해능, 항돌연변이효과 등에 대한 생리활성 기능을 보였으며, 기존의 에탄올이나 열수 추출법에 비하여 유사하거나 더 높은 생리활성 효과가 나타나기도 했다.

• 한국환경성돌연변이발암원학회지
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• 제16권1호
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• pp.24-29
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• 1996

In this study, the micronucleus test with peripheral blood using acridine orange coated slides was evaluated in mice treated with mitomycin C(MMC) at doses of 0.5, 1.0 and 1.5 mg/kg body weight. The peripheral bloods were obtained at 0, 24, 48 and 72h after treatment. The frequencies of micronucleated reficulocytes(MNRET) in the MMC-treated groups increased dose-dependently, and showed a peak time at 48h after treatment. We also performed the sex differences of MNRET frequency in 0.5 mg/kg MMC treated group, and we observed no sex differences in this experiment. And we evaluated the usefulness of a direct acting clastogen, N-methyl-N-nitrosourea and a indirect acting clastogen, benzo(a) pyrene as the positive control in this supravital micronucleus test. They also caused a significant increase in MNRET frequencies. These results suggest that the supravital staining micronucleus test using MNRET can be useful tool to evalulate the quantitative and qualitative assessment of genotoxicity in vivo compared to classical in vivo micronucleus test using bone-marrow cells.