• Title/Summary/Keyword: comet assay-chromosomal

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Genotoxicity Studies on Carrageenan: Short-term In Vitro Assays

  • Chung, Young-Shin;Eum, Ki-Hwan;Choi, Seon-A;Oh, Se-Wook;Park, Sue-Nie;Yum, Young-Na;Kim, Joo-Hwan;Seo, Young-Rok;Lee, Michael
    • Toxicological Research
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    • v.25 no.1
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    • pp.51-58
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    • 2009
  • Carrageenan is a naturally-occurring sulfated polygalactan which has been widely used in the dairy industry and a gelling agent in non-dairy products. In this study, four short-term in vitro genotoxicity assays were investigated to evaluate the potential genotoxic effects of carrageenan. The mutagenicity of carrageenan was evaluated up to a maximum dose of 5 mg/plate in Ames test. There was no increase in the number of revertant colonies compared to its negative control at any dose in all of strains tested. To assess clastogenic effect, the in vitro chromosomal aberration assay was performed using Chinese hamster lung cells. Carrageenan was not considered to be clastogenic in this assay at up to the highest feasible concentration which could be evaluated. The in vitro comet assay and micronucleus test results obtained on L5178Y cells also revealed that carrageenan has no genotoxicity potential, although there was a marginal increase in micronuclei frequencies and DNA damage in the respective micronucleus and comet assays. Taken together, our results indicate that carrageenan was not genotoxic based on four in vitro genotoxicity results.

Antioxidant Activity of Vegetables or Fruits Extract in Mice (야채 및 과일추출물의 항산화작용)

  • Heo Chan;Kim Nam Yee;Kim Hyun Pya;Heo Moon Young
    • YAKHAK HOEJI
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    • v.49 no.3
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    • pp.249-254
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    • 2005
  • The ethanol extracts of the mixed vegetables (Bioactive Vegetables, BV) and the mixed fruits (Bioactive Fruits, BF) were evaluated for their in vivo antioxidant activities. Four weeks treatment of oral administration was performed to mice. A $KBrO_3$ as a potent oxidant was used to induce the oxidative stress for in vivo experiment. BV and BF were shown to possess the significant inhibitory effect of lipid peroxidation as measured by the level of malondialdehyde (MDA) formation although the potencies were not higher than those of well-known antioxidants such as vitamin C, trolox and quercetin. Furthermore, BV and BF inhibited DNA damage assessed by single cell gel electrophoresis (comet assay) and reduced the micronucleated reticulocyte (MNRET) formation of peripheral blood. Antioxidants tested also revealed potent inhibitory activities higher than BV and BF. These antigenotoxic activity profiles were similar to that of abovementioned inhibition of lipid peroxidation. Therefore, BV and BF having mild antioxidant activity as functional food candidates may be useful natural antioxidants by the inhibiting of lipid peroxidation and the protecting oxidative DNA and chromosomal damage.

Genotoxicity Study on Khal, a Halocidin Derivative, in Bacterial and Mammalian Cells

  • Kim, Youn-Jung;Kim, Mi-Soon;Jeon, Hee-Kyoung;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.2 no.3
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    • pp.151-158
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    • 2006
  • Khal was a synthetic congener of halocidin, a heterodimeric peptide consisting of 19 and 15 amino acid residues detected in Halocynthia aurantium. This compound was considered a candidate for the development of a novel peptide antibiotic. The genotoxicity of Khal was subjected to high throughput toxicity screening (HTTS) because they revealed strong antibacterial effects. Mouse lymphoma thymidine kinase ($tk^{+/-}$) gene assay (MOLY), single cell gel electrophoresis (Comet) assay and chromosomal aberration assay in mammalian cells and Ames reverse mutation assay in bacterial system were used as simplified, inexpensive, short-term in vitro screening tests in our laboratory. These compounds are not mutagenic in S. typhimurium TA98 and TA100 strains both in the presence and absence of metabolic activation. Before performing the comet assay, $IC_{20}$ of Khal was determined the concentration of $25.51\;{\mu}/mL\;and\;21.99\;{\mu}g/mL$ with and without S-9, respectively. In the comet assay, Khal was not induced DNA damage in mouse lymphoma cell line. Also, the mutation frequencies in the Khal-treated cultures were similar to the vehicle controls. It is suggests that Khal is non-mutagenic in MOLY assay. And no clastogenicity was observed in Khal-treated Chinese hamster lung cells. The results of this battery of assays indicate that Khal has no genotoxic potential in bacterial or mammalian cell systems. Therefore, we suggest that Khal, as the optimal candidates with both no genotoxic potential and antibacterial effects must be chosen.

Genotoxicity Assessment of Gardenia Yellow using Short-term Assays

  • Chung, Young-Shin;Eum, Ki-Hwan;Ahn, Jun-Ho;Choi, Seon-A;Noh, Hong-June;Seo, Young-R.;Oh, Se-Wook;Lee, Michael
    • Molecular & Cellular Toxicology
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    • v.5 no.3
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    • pp.257-264
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    • 2009
  • Gardenia yellow, extracted from gardenia fruit, has been widely used as a coloring agent for foods, and thus, safety of its usage is of prime importance. In the current study, short-term genotoxicity assays were conducted to evaluate the potential genotoxic effects of gardenia yellow. The gardenia yellow used was found to contain 0.057 mg/g of genipin, a known biologically active compound of the gardenia fruit extract. Ames test did not reveal any positive results. No clastogenicity was detected by a chromosomal aberration test, even on evaluation at the highest feasible concentration of gardenia yellow. Gardenia yellow was also shown to be non-genotoxic using an in vitro comet assay and a micronucleus test with L5178Y cells, although a marginal increase in DNA damage and micronuclei frequency was reported in the respective assays. Additionally, in vivo micronucleus test results clearly demonstrated that oral administration of gardenia yellow did not induce micronuclei formation in the bone marrow cells of male ICR mice. Taken together, our results indicate that gardenia yellow is not mutagenic to bacterial cells, and that it does not cause chromosomal damage in mammalian cells, either in vitro or in vivo.

Genotoxicity Study of Sophoricoside in Bacterial and Mammalian Cell System

  • Yun, Hye-Jung;Kim, Youn-Jung;Kim, Eun-Young;Kim, Youngsoo;Kim, Mi-Kyung;Lee, Seung-Ho;Jung, Sang-Hun;Ryu, Jae-Chun
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.05a
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    • pp.183-184
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    • 2003
  • Sophoricoside was isolated as the inhibitor of IL-5 bioactivity from Sophora japonica (Leguminosae). It has been reported to have an anti-inflammatory effect on rat paw edema model. To develop as an anti-allergic drug, genotoxicity of sophoricoside was investigated in bacterial and mammalian cell system such as Ames bacterial test, chromosomal aberration assay, Comet assay and MOLY assay. In Ames test, sophoricoside of 5000 ∼ 313 $\mu\textrm{g}$/plate concentrations was not shown significant mutagenic effect in Salmonella typhimurium TA98, TA100, TA1535 and TA1537 strains. The cytotoxicity (IC$\_$50/ and IC$\_$20/) of sophoricoside was determined above the concentration of 5000 $\mu\textrm{g}$/ml in Chinese hamster lung (CHL) fibroblast cell and L5178Y mouse lymphoma cell line. At concentrations of 5000, 2500 and 1250 $\mu\textrm{g}$/ml, this compound was not induced chromosomal aberration in CHL fibroblast cell in the absence and presence of S-9 metabolic activation system. Also in comet assay, DNA damage was not observed in L5178Y cell line. Also in MOLY assay, sophoricoside of 5000 ∼ 313 $\mu\textrm{g}$/ml concentrations was not shown significant mutagenic effect in absence of S-9 metabolic activation system. However, the higher concentration of 5000 and 2500 $\mu\textrm{g}$/ml of sophoricoside induced the increased mutation frequency (MF) in the presence of S-9 metabolic activation system. From these results, no genotoxic effects of sophoricoside observed in bacterial systems whereas, genotoxic effects observed in mammalian cell systems in the presence of metabolic activation system. These results suggested that the metabolite(s) of sophoricoside can cause some genotoxic effects in mammalian cells.

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Genotoxic Effect of Air-borne Particulate Matter in Residential Area of Seoul City (서울시내 주거지역 미세먼지의 유전독성 영향)

  • Oh Seung Min;Sung Hye Kyoung;Kim Eun Sil;Kim Jong Geuk;Ryu Byung Taek;Chung Kyu Hyuck
    • Environmental Analysis Health and Toxicology
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    • v.20 no.4 s.51
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    • pp.365-374
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    • 2005
  • Ambient air particulate matters are classified into two distinct modes in sire distribution, namely the coarse and fine particles. Correlation between high particulate concentration and adverse effect on human populations has long been recognized. However, the toxicology of these adverse efforts has not been clarified. We investigated the genotoxic effect of PM 2.5 collected from urban area in Seoul by comet assay (A549 cells), CBMN assay (CHO-K1 cells) and EROD-microbioassay (H4IIE cells). Results from in vitro micronucleus assay and comet assay showed that PM 2.5 samples collected from traffic area, residential area and indoor air induced chromosomal damage and DNA breakage in a non-cytotoxic dose. The complex mixture effect of these PM 2.5 extracts was quantified by EROD-microbioassay in terms of its bio-TEQ (biologiral -TCDD equivalent concentration) which was 70.87$\pm$28.07, 93.55$\pm$21.80 and 14.31 $\pm$ 1.10 ng/g-PM 2.5 in traffic area, residental area and indoor air samples, respectively. Conclusively, we suggested that PM 2.5 collected from traffic area and residential area contains CYPIA inducer and genotoxic materials.

Study on Genotoxicity of Crocin, a Component of Gardenia Fruit, in Bacterial and Mammalian Cell Systems

  • Choi, Hae-Yeon;Kim, Youn-Jung;Jeon, Hee-Kyung;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.4 no.4
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    • pp.285-292
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    • 2008
  • Crocin is one of the major components of gardenia fruit and saffron which are widely used as natural food colorants and as traditional Chinese medicines. However, the genotoxicity data on crocin are not sufficient for safety evaluation. The purpose of this study was the examination of the genotoxicity on crocin from gardenia yellow in bacterial and mammalian cells, using various genotoxic battery testing assays and the influence of crocin on methyl methanesulfonate (MMS) and ${H_2}{O_2}$-induced DNA damage in vitro, using single cell gel electrophoresis (comet) assay. From results, no considerable mutagenicity and clastogenicity were seen in bacteria and mammalian cells treated with crocin, by Ames test, chromosomal aberration assay, ${tk}^{+/-}$ gene forward mutation assay and comet assay. And, post-treatment with crocin significantly suppressed ${H_2}{O_2}$-induced DNA damage in a dose-dependent manner. In conclusion, the findings of the present study and other previous observations indicate that crocin has no genotoxic potential. And it showed that crocin clearly repressed the genotoxic potency of ${H_2}{O_2}$. These results suggest that anti-oxidative effects of crocin may be involved in the protective effects of DNA damage.

Evaluation of in vivo Genotoxicity of Plant Flavonoids, Quercetin and Isoquercetin (식물유래 플라보노이드 Quercetin과 Isoquercetin의 생체 내 유전독성평가)

  • Pak, Bumsoo;Han, Sehee;Lee, Jiyeon;Chung, Young-Shin
    • Journal of Food Hygiene and Safety
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    • v.31 no.5
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    • pp.356-364
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    • 2016
  • In vivo genotoxic potential of isoquercetin, a plant common flavonoid, in comparison with quercetin was investigated for the DNA breakage and the clastogenicity endpoints. Male ICR mice were administered by oral gavage for 3 days with $3{\times}0.5%$ carboxymethyl cellulose (CMC), 3 ${\times}$ isoquercetin (250, 500 mg/kg/day), 3 ${\times}$ quercetin (250, 500 mg/kg/day) and 2 ${\times}$ ethyl methanesulfonate (EMS, 200 mg/kg/day). Tissues were collected 48 hours after the first treatment and within 3 hours after the last treatment. The DNA damages were evaluated using Comet assay in liver and stomach, while the clastogenicities were determined using micronucleus test in bone marrow of same animals. The treatment of isoquercetin as well as quercetin did not cause the DNA damages in liver and stomach, and not induce the frequencies of micronucleus polychromatic erythrocytes in bone marrow. In conclusion, isoquercetin as well as quercetin did not cause the DNA breakages and the chromosomal damages in vivo system in these study conditions.

GENOTOXICITY STUDY OF SOPHORICOSIDE IN BACTERIAL AND MAMMALIAN CELL SYSTEM

  • Kim, Youn-Jung;Kim, Yun-Hwa;Park, Hyo-Joung;Gil, Ji-Suk;Kim, Young-Soo;Kim, Mi-Kyung;Lee, Seung-Ho;Jung, Sang-Hun;Park, Jong-Bum;Kim, Tae-Hwam;Ryu, Jae-Chun
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2001.10a
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    • pp.182-182
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    • 2001
  • Sophoricoside was isolated as the inhibitor of IL-5 bioactivity from Sophora japonica (Leguminosae). It has been reported to have an anti-inflammatory effect on rat paw edema model. To develope as an anti-allergic drug, genotoxicity of sophoricoside was investigated in bacterial and mammalian cell system such as Ames bacterial test, chromosomal aberration assay and single cell gel electrophoresis (Comet) assay.(omitted)

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Effects of 835-MHz Radiofrequency Radiation on the Chromosomal DNA of Mouse Thymic Lymphoma L5178Y $Tk^{+/-}$ Cells

  • Choi, Jong-Soon;Son, TaeHo;Chang, Sung-Keun;Hong, Sae-Yong
    • Korean Journal of Environmental Biology
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    • v.22 no.4
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    • pp.507-512
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    • 2004
  • This study was focused on the risk assessment of whether radiofrequency electromagnetic fields generated by mobile phone is cytogenetically toxic or not. We conducted the effects of 835-MHz electromagnetic field (EMF) on DNA strand breaks in mouse thymic lymphoma L5178Y $Tk^{+/1-}$ cells using alkaline comet assay. EMF frequency 835-MHz we chosen is one of the most popular communication frequency bands in Korean code-division multiple-access (CDMA) mobile phone system. The cells were exposed to 835-MHz EMF alone or 835-MHz EMF combined with cyclophosamide(CPA) or 4-nitroquinoline-1-oxide (4NQO) at specific absorption rate (SAR) of 4.0 W $kg^{-l}$ for 24 and 48hrs. DNA damage expressed as tail moment was increased more than two-fold after exposure to 835-MHz EMF for 24 and 48hr. In particular, CPA for 48hr and 4NQO for 24 hr enhanced notably the tail moment to 9-fold and 16-fold in the presence of 835-MHz EMF, respectively, compared to each single treatment. From these results, it appears that exposure to CDMA-mobile phone radiation at 835-MHz frequency may potentiate DNA strand breaks of mouse thymic lymphoma L5178Y $Tk^{+/1-}$;cells under the defined conditions of this study.