• Title/Summary/Keyword: DNA single-strand breaks

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Antigenotoxic Effect of Dominant Bacterial Isolates from Kimchi in vitro (김치로부터 분리된 우세 균주들의 in vitro 항유전 독성효과)

  • Choi, Jun-Won;Park, Jong-Heum;Ji, Seung-Taek;Choi, Ok-Byung;Shin, Heuyn-Kil
    • Korean Journal of Food Science and Technology
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    • v.31 no.4
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    • pp.1071-1076
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    • 1999
  • This study was carried out to isolate the colonies of dominant fermented bacteria from Kimchi (Korean native fermented foodstuffs) and investigate their inhibitory potentials on mutagenesis and carcinogenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) as direct carcinogen. For this purpose, single cell gel electrophoresis technique (SCGE assay, or comet assay) which is a sensitive and rapid technique for detecting the presence of DNA strand breaks in individual cells was used. DNA damages of Kimchi isolates were compared with that of the positive control, MNNG. Among 3 isolates from Tongbaechu Kimchi, two isolates B-1 and B-2 showed antigenotoxicities (p<0.01). All 4 isolates from Yulmu Kimchi had antigenotoxicities (p<0.05). Also, 3 of 5 isolates from Chonggak Kimchi and 2 of 9 isolates from Kaktugi were antigenotoxic (p<0.05 and p<0.01, respectively).

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Genotoxicity and Identification of Differentially Expressed Genes of Formaldehyde in human Jurkat Cells

  • Kim, Youn-Jung;Kim, Mi-Soon;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.1 no.4
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    • pp.230-236
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    • 2005
  • Formaldehyde is a common environmental contaminant found in tobacco smoke, paint, garments, diesel and exhaust, and medical and industrial products. Formaldehyde has been considered to be potentially carcinogenic, making it a subject of major environmental concern. However, only a little information on the mechanism of immunological sensitization and asthma by this compound has been known. So, we performed with Jurkat cell line, a human T lymphocyte, to assess the induction of DNA damage and to identify the DEGs related to immune response or toxicity by formaldehyde. In this study, we investigated the induction of DNA single strand breaks by formaldehyde using single cell gel electrophoresis assay (comet assay). And we compared gene expression between control and formaldehyde treatment to identify genes that are specifically or predominantly expressed by employing annealing control primer (ACP)-based $GeneFishing^{TM}$ method. The cytotoxicity ($IC_{30}$) of formaldehyde was determined above the 0.65 mM in Jurkat cell in 48 h treatment. Based on the $IC_{30}$ value from cytotoxicity test, we performed the comet assay in this concentration. From these results, 0.65 mM of formaldehyde was not revealed significant DNA damages in the absence of S-9 metabolic activation system. And the one differentially expressed gene (DEG) of formaldehyde was identified to zinc finger protein 292 using $GeneFishing^{TM}$ method. Through further investigation, we will identify more meaningful and useful DEGs on formaldehyde, and then can get the information on the associated mechanism and pathway with immune response or other toxicity by formaldehyde exposure.

Evaluation of Oxidative DNA Damage Using an Alkaline Single Cell Gel Electrophoresis (SCGE) Comet Assay, and the Protective Effects of N-Acetylcysteine Amide on Zearalenone-induced Cytotoxicity in Chang Liver Cells

  • Kang, Changgeun;Lee, Hyungkyoung;Yoo, Yong-San;Hah, Do-Yun;Kim, Chung Hui;Kim, Euikyung;Kim, Jong Shu
    • Toxicological Research
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    • v.29 no.1
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    • pp.43-52
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    • 2013
  • Zearalenone (ZEN) is a non-steroidal estrogenic mycotoxin produced by several species of Fusarium that are found in cereals and agricultural products. ZEN has been implicated in mycotoxicosis in farm animals and in humans. The toxic effects of ZEN are well known, but the ability of an alkaline Comet assay to assess ZEN-induced oxidative DNA damage in Chang liver cells has not been established. The first aim of this study was to evaluate the Comet assay for the determination of cytotoxicity and extent of DNA damage induced by ZEN toxin, and the second aim was to investigate the ability of N-acetylcysteine amide (NACA) to protect cells from ZEN-induced toxicity. In the Comet assay, DNA damage was assessed by quantifying the tail extent moment (TEM; arbitrary unit) and tail length (TL; arbitrary unit), which are used as indicators of DNA strand breaks in SCGE. The cytotoxic effects of ZEN in Chang liver cells were mediated by inhibition of cell proliferation and induction of oxidative DNA damage. Increasing the concentration of ZEN increased the extent of DNA damage. The extent of DNA migration, and percentage of cells with tails were significantly increased in a concentration-dependent manner following treatment with ZEN toxin (p < 0.05). Treatment with a low concentration of ZEN toxin (25 ${\mu}M$) induced a relatively low level of DNA damage, compared to treatment of cells with a high concentration of ZEN toxin (250 ${\mu}M$). Oxidative DNA damage appeared to be a key determinant of ZEN-induced toxicity in Chang liver cells. Significant reductions in cytolethality and oxidative DNA damage were observed when cells were pretreated with NACA prior to exposure to any concentration of ZEN. Our data suggest that ZEN induces DNA damage in Chang liver cells, and that the antioxidant activity of NACA may contribute to the reduction of ZEN-induced DNA damage and cytotoxicity via elimination of oxidative stress.

Decrease of Genotoxicity by Red Ginseng Root Extract (I) - Decrease of UV -induced Genotoxicity by Red Ginseng Root Extract in Cultured NIH3T3 Cells (홍삼 추출물에 의한 유전독성 감소효과 (I) - 배양 NIH3T3 세포에서 자외선에 의한 유전독성의 감소에 미치는 홍삼추출물 처리효과)

  • 김완주;유병수
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.24 no.1
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    • pp.74-86
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    • 1998
  • We have studied the effects of red ginseng root extract on the decrease of UV-induced genotoxicity in cultured NIH3T3 cells. The increase in survival and the recovery from DNA synthesis inhibition in UV-irradiated cells as a function of normal medium incubation time was potentiated by the presence of the ginseng extract. The extract also increased the UV-induced excision repair as determined by unscheduled DNA synthesis. The amount of UV-induced DNA single strand breaks that are accumulated by polymerase inhibitors was significantly increased by the presence of the extract. These results suggest that the red ginseng extract activates the incision/excision step of UV-induced repair and could be used as a reagent for protecting UV-induced genotoxicity and cytotoxicity.

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Assessment of DNA Damage using an Alkaline Single Cell Gel Electrophoresis (SCGE) Comet Assay and Toxic Effects in Chickens by T-2 Toxin Treatment (T-2 toxin을 투여한 닭에서 Comet assay 방법을 이용한 DNA 손상 평가와 독성)

  • Hah Dae-Sik;Heo Jung-Ho;Lee Kuk-Cheon;Cho Myung-Heui;Kim Kuk-Hun;Kim Chung-Hui;Lue Jae-Du;Lee Seung-Hwan;Kim Gon-Sup;Kim Eui-Gyung;Kim Jong-Shu
    • Toxicological Research
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    • v.22 no.2
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    • pp.75-85
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    • 2006
  • This study was designed to evaluate the possible DNA damaging effects of T-2 toxin using an alkaline single cell gel electrophoresis (SCGE) comet assay and also to investigate toxic effects in chickens. A total of 20 chickens were used in these experiments. Graded concentrations of dietary T-2 toxin (0, 4, 8, and $16{\mu}g/g$ of diet) were given to groups of 5 broiler chickens. In comet assay, The DNA damage was analysed by the tail extent moment (TEM) and tail length (TL), which were used as markers of DNA strand breaks in SCGE. A significant dose-dependent increase in the extent of DNA migration as well as in the percentage of cells with tails was observed after treatment with T-2 toxin (P<0.05). Treatment with the low T-2 toxin ($4{\mu}/g$ of diet) induced a relatively low level of DNA damage in comparison with the high T-2 toxin ($16{\mu}/g$ of diet) group. The growth rate was significantly reduced by concentrations of 8, and $16{\mu}/g$ of diet (P < 0.05). The feed conversion ratio were significantly affected by any concentrations (P < 0.05). The relative weight of the spleen, and lung was decreased by the growth inhibitory concentrations. The bursa of Fabricius, thymus, and kid- ney were decreased in relative weight by concentrations of $16{\mu}/g$ of diet. The relative weight of the liver and heart were unaffected. The hemoglobin (Hb), hematocrit (HCT), and mean corpuscular hemoglobin (MCH) were decreased at concentration of $16{\mu}/g$ of diet. As compared with control chickens, there was no marked change in serum components except uric acid in T-2 treated chickens. All lymphoid tissues retained atrophic and lymphoid cell depletion throughout the three weeks trial.

Genotoxicity Study of sophoricoside derivatives in mammalian cells system

  • Yun, Hye-Jung;Kim, Youn-Jung;Kim, Eun-Young;Jung, Sang-Hun;Kim, Youngsoo;Kim, Mi-Kyung;Lee, Seung-Ho;Ryu, Jae-Chun
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.05a
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    • pp.185-185
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    • 2003
  • To develope the novel anti-allergic drug, many sophoricoside derivatives were synthesized. Among these derivatives, JSH-II-3, JSH-Ⅵ-3, JSH-Ⅶ-3, and JSH-Ⅷ-3 were selected and subjected to high throughput toxicity screening (HTTS) because they revealed strong IL-5 inhibitory activity and limitation of quantity. Mouse lymphoma thymidine kinase (tk$\^$+/-/) gene assay (MOLY) and single cell gel electrophoresis (Comet) assay in mammalian cells were used as HTTS tool in our laboratory. In MOLY assay, JSH-Ⅶ-3 at 50 ∼ 6 $\mu\textrm{g}$/ml concentrations was not shown significant mutagenic effect in the absence and presence of S-9 metabolic activation system. However, the concentration of ISH-II-3, 38 $\mu\textrm{g}$/ml, induced increased mutation frequency (MF) in the presence of S-9 metabolic activation system. Also in comet assay, DNA damage was not observed in JSH-Ⅵ-3 and JSH-Ⅶ-3, wherase concentration of 32.8 $\mu\textrm{g}$/ml in JSH-II-3 and 13.9 $\mu\textrm{g}$/ml in JSH-Ⅶ-3 were induced DNA damage in the absence of S-9 metabolic activation system. Therefore, we suggest that JSH-Ⅵ-3 and JSH-Ⅶ-3 have no genotoxic effects but JSH-II-3 and JSH-Ⅷ-3 induce some mutagenicity and DNA strand breaks in mouse lymphoma cell line used this study.

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Selective Effects of Curcumin on CdSe/ZnS Quantum-dot-induced Phototoxicity Using UVA Irradiation in Normal Human Lymphocytes and Leukemia Cells

  • Goo, Soomin;Choi, Young Joo;Lee, Younghyun;Lee, Sunyeong;Chung, Hai Won
    • Toxicological Research
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    • v.29 no.1
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    • pp.35-42
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    • 2013
  • Quantum dots (QDs) have received considerable attention due to their potential role in photosensitization during photodynamic therapy. Although QDS are attractive nanomaterials due to their novel and unique physicochemical properties, concerns about their toxicity remain. We suggest a combination strategy, CdSe/ZnS QDs together with curcumin, a natural yellow pigment from turmeric, to reduce QD-induced cytotoxicity. The aim of this study was to explore a potentially effective cancer treatment: co-exposure of HL-60 cells and human normal lymphocytes to CdSe/ZnS QDs and curcumin. Cell viability, apoptosis, reactive oxygen species (ROS) generation, and DNA damage induced by QDs and/or curcumin with or without ultraviolet A (UVA) irradiation were evaluated in both HL-60 cells and normal lymphocytes. In HL-60 cells, cell death, apoptosis, ROS generation, and single/double DNA strand breaks induced by QDs were enhanced by treatment with curcumin and UVA irradiation. The protective effects of curcumin on cell viability, apoptosis, and ROS generation were observed in normal lymphocytes, but not leukemia cells. These results demonstrated that treatment with QD combined with curcumin increased cell death in HL-60 cells, which was mediated by ROS generation. However, curcumin acted as an antioxidant in cultured human normal lymphocytes.

Evaluation of DNA Damage by Mercury Chloride (II) and Ionizing Radiation in HeLa Cells (이온화 방사선 및 염화수은(II)에 의한 자궁경부암 세포의 DNA 손상 평가)

  • Woo Hyun-Jung;Kim Ji-Hyang;Antonina Cebulska-Wasilewska;Kim Jin-Kyu
    • Korean Journal of Environmental Biology
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    • v.24 no.1 s.61
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    • pp.46-52
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    • 2006
  • The mercury is among the most highly bioconcentrated toxic trace metals. Many national and international agencies and organisations have targeted mercury for the possible emission control. The mercury toxicity depends on its chemical form, among which alkylmercury compounds are the most toxic. A human cervix uterus cancer cell line HeLa cells was employed to investigate the effect of the toxic heavy metal mercury (Hg) and ionizing radiation. In the in vitro comet assays for the genotoxicity in the HeLa cells, the group of Hg treatment after irradiation showed higher DNA breakage than the other groups. The tail extent moment and olive tail moment of the control group were $4.88{\pm}1.00\;and\;3.50{\pm}0.52$ while the values of the only Hg treatment group were $26.90{\pm}2.67\;and\;13.16{\pm}1.82$, respectively. The tail extent moment and olive tail moment of the only 0.001, 0.005, 0.01 Hg group were $12.24{\pm}1.82,\;8.20{\pm}2.15,\;20.30{\pm}1.30,\;12.26{\pm}0.52,\;40.65{\pm}2.94\;and \;20.38{\pm}1.49$, respectively. In the case of Hg treatment after irradiation, the tail extent moment and olive tail moment of the 0.001, 0.005, 0.01 Hg group were $56.50{\pm}3.93,\;32.69{\pm}2.48,\;62.03{\pm}5.14,\;31.56{\pm}1.97,\;72.73{\pm}3.70\;and \;39.44{\pm}3.23$, respectively. The results showed that Hg induced DNA single-strand breaks or alkali labile sites as assessed by the Comet assay. It is in good agreement with the reported results. The mercury inhibits the repair of DNA. The bacterial formamidopyrimidine-DNA glycosylase (Epg protein) recognizes and removes some oxidative DNA base modifications. Enzyme inactivation by Hg (II) may therefore be due either to interactions with rysteine residues outside the metal binding domain or to very high-affinity binding of Hg (II) which readily removes Zn (II) from the zinc finger.

Responses of Various Biomarkers in Common Carp (Cyprinus carpio) Exposed to Benzo[k]fluoranthene

  • Kim, Woo-Keun;Kim, Ja-Hyun;Yeom, Dong-Hyuk;Lee, Sung-Kyu
    • Korean Journal of Ecology and Environment
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    • v.41 no.3
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    • pp.331-337
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    • 2008
  • Polycyclic aromatic hydrocarbons (PAHs) derived from leakage of fossil fuels and incomplete combustion of organic materials have been considered as harmful contaminants in environments. This study evaluated the effect of benzo[k]fluoranthene (BkF), one of the PAHs, using the multiple biomarkers and applied the integration model with those biomarker responses. After 10 days of the exposure at the measured concentrations of BkF (6, 25, and 45 ${\mu}g\;L^{-1}$), the changes of the four biomarkers, that is, 7-ethoxyresorufin-O-deethylase (EROD), DNA single-strand breaks (Comet), acetylcholinesterase (AChE) and vitellogenin (VTG) in the common carp (Cyprinus carpio) were observed. The standardized values of four biomarker responses were computed and integrated as star plots, representing Integrated Biomarker Respnse (IBR) values. DNA damage was induced in a dose-dependent manner, and increased significantly compared with that in the control. EROD and VTG levels were significantly elevated at low concentrations of BkF. On the other hand, AChE activities were not altered by BkF. IBR values increased as the exposure concentrations increased. Thus, the metabolic, endocrine and genetic changes of the biomarker responses in the common carp exposed to BkF should be considered in the case of the ecological risk assessment of the BkF in fish and it can be used as a biomonitoring tool in aquatic ecosystems. In addition, star plots can be used as a useful analysis tool in multibiomarker integration approach.