• Title, Summary, Keyword: DNA Damage

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Use of comet assay as a bioassay in marine organisms exposed to genotoxicants (유전독성물질로 오염된 해양생물의 생물검정법으로서 comet assay 이용)

  • Kim Gi-Beum;An Joon-Gun;Kim Jae-Won
    • Journal of Environmental Science International
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    • v.14 no.11
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    • pp.1071-1079
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    • 2005
  • Using single cell gel electrophoresis, DNA single strand breaks were determined in various marine organisms. DNA damage on fish blood cells was detected to know whether there was a difference between Incheon, Pohang, Masan, and Tongyeong as a control site. Tongyeong showed the lowest DNA damage among the study areas. Mussels, transplanted to Masan Bay for one month, revealed high DNA damage at sites with high economical activity. In two weeks exposure of polychaete to Incheon sediments, higher DNA damage was detected in the sediment adjacent to Incheon harbor than open sea. These results suggested that the marine organism from the polluted area revealed a relatively high DNA damage. In addition, these areas might be contaminated with genotoxic compounds and comet assay was useful as a bioassay to detect DNA damage in marine organisms.

The DNA Damage by Fish Oil Perokidation Products 1. DNA Damage by the Peroxidation Products of Total Lipid Fraction Extracted from Mackerel (어유산화생성물의 DNA 손상작용 1. 총지질산화생성물의 DNA 손상작용)

  • KANG Jin-Hoon;BYUN Han-Seok;LEE Yong-Woo;KIM Seon-Bong;PARK Young-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.20 no.3
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    • pp.213-218
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    • 1987
  • The DNA damage mechanism by fish oil peroxidation was investigated through the model system of a DNA-mackerel lipid at $37^{\circ}C$. Mackerel lipid peroxidation products induced a great DNA damage with the increment of its concentration, and such DNA damage in all systems examined occurred below $100millieq{\cdot}/kg$ in POV (peroxide value) Singlet oxygen $(^1O_2)$ and superoxide anion${\cdot}O_2^-$ greatly participated in the DNA damage during peroxidation of mackerel lipid, while hydrogen peroxide$(H_2O_2)$ and hydroxyl radical $({\cdot}OH)$ did little show the DNA damage. From the results of the addition of several active oxygen scavengers to the DNA-lipid systems, singlet oxygen ana superoxide anion greatly affected to the increase of POV ana to the DNA damage by mackerel lipid peroxidation, respectively. It indicates that there was a close relationship between the effects of active oxygens in the mackerel lipid peroxidation and its DNA damage mechanism.

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Role of Active Oxygens on DNA Damage by Low Molecular Carbonyl Compounds Derived from Maillard Reaction (Maillard 반응에서 유래되는 저분자 Carbonyl 화합물의 DNA손상작용에 대한 활성산소종의 역할)

  • 김선봉;박성준;강진훈;변한석;박영호
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.19 no.6
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    • pp.565-570
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    • 1990
  • The role of the active oxygens on plasmid DNA damage by carbonyl compounds derived from Maillard reaction was investigated. Plasmid DNA extracted from E. coli Hb1O1 was reacted with carbonyl compounds, such as glyoxal, methyl glyoxal, dihydroxyacetone, diacetyl, glyceraldehyde, glycolaldehyde and furfural with and without the active oxygen scavengers at 37$^{\circ}C$ for 6 hours, and then the degree of damage was determined by using 1 % agarose gel electro-phoresis. All of the carbonyl compounds except furfural caused to damage of DNA. Among these, glyoxal, methyl glyoxal and dihydroxyacetone markedly induced the damage of DNA. On the other hand, the DNA damage by the carbonyl compounds was greatly inhibited by catalase, superoxide dismutase and $\alpha$-tocopherol it is considered that the damage of DNA is due to active oxygens, such as singlet oxygen, hydrogen peroxide and superoxide anion generated during the autoxidation of carbonyl compounds.

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Ser1778 of 53BP1 Plays a Role in DNA Double-strand Break Repairs

  • Lee, Jung-Hee;Cheong, Hyang-Min;Kang, Mi-Young;Kim, Sang-Young;Kang, Yoon-Sung
    • The Korean Journal of Physiology and Pharmacology
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    • v.13 no.5
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    • pp.343-348
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    • 2009
  • 53BP1 is an important genome stability regulator, which protects cells against double-strand breaks. Following DNA damage, 53BP1 is rapidly recruited to sites of DNA breakage, along with other DNA damage response proteins, including ${\gamma}$-H2AX, MDC1, and BRCA1. The recruitment of 53BP1 requires a tandem Tudor fold which associates with methylated histones H3 and H4. It has already been determined that the majority of DNA damage response proteins are phosphorylated by ATM and/or ATR after DNA damage, and then recruited to the break sites. 53BP1 is also phosphorylated at several sites, like other proteins after DNA damage, but this phosphorylation is not critically relevant to recruitment or repair processes. In this study, we evaluated the functions of phosphor-53BP1 and the role of the BRCT domain of 53BP1 in DNA repair. From our data, we were able to detect differences in the phosphorylation patterns in Ser25 and Ser1778 of 53BP1 after neocarzinostatin-induced DNA damage. Furthermore, the foci formation patterns in both phosphorylation sites of 53BP1 also evidenced sizeable differences following DNA damage. From our results, we concluded that each phosphoryaltion site of 53BP1 performs different roles, and Ser1778 is more important than Ser25 in the process of DNA repair.

The DNA Damage of Fish Oil Peroxidation Products 2. DNA Damage by the Peroxidation Products of Polar and Non-polar Lipid Fractionated from Mackerel Lipid (어유산화생성물의 DNA손상작용 2. 극성 및 비극성지질획분산화생성물의 DNA 손상작용)

  • KANG Jin-Hoon;Do Jung-Roung;KIM In-Soo;KIM Seon-Bong;PARK Young-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.20 no.4
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    • pp.300-307
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    • 1987
  • The present study was investigated on the DNA damage by the peroxidation of polar and non-polar lipid fractionated from mackerel lipid to elucidate the DNA damage mechanism by fish oil peroxidation. The degree of DNA damage by polar lipid peroxidation became greater with the increase of its concentration, and such DNA damage was induced below 100 millieq./kg in POV for 4 days incubation. Among the polar lipid peroxidation products, singlet oxygen $^1O_2$ and superoxide anion ${\cdot}O_2^-$ greatly affected to the DNA damage than hydrogen peroxide $H_2O_2$ and hydroxyl radical ${\cdot}OH$. Non-polar lipid peroxidation also induced the DNA damage with the increase of its concentration, but such effect was lower than the case of total lipid and polar lipid. And, the effects of active crygens on the DNA damage by non-polar lipid peroxidation was the same as in the case of total and polar lipid peroxidation.

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Antioxidant Activities and Inhibitory Effect on Oxidative DNA Damage of extracts from Abeliophylli distichi Folium (미선나무 잎 추출물의 항산화 및 산화적 DNA 손상억제 활성)

  • Park, Jae-Ho
    • The Korea Journal of Herbology
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    • v.26 no.4
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    • pp.95-99
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    • 2011
  • Objective : In this study, we demonstrate the protective effect on oxidative DNA damage of leaf extracts from Abeliophylli distichi Folium via its antioxidant activity for the establishment of new value for the herbal medicine. Methods : Abeliophylli distichi Folium leaves were extracted with hot-water and ethylacetate (EtOAC). The 1,1-diphenyl-2- picrylhydrazyl (DPPH) radical and hydroxyl scavenging assay and $Fe^{2+}$ chelating assay were performed for antioxidative effect and ${\varphi}$X-174 RF I DNA cleavage assay and intracellular DNA damage assay were used for inhibitory effect of intracellular DNA damage. Results : In DPPH, Hydroxyl radical scavenging activity and $Fe^{2+}$ chelating activity of EtOAC extracts were 94.72%, 62.88%, 41.13%, and hot-water extracts were 88.86%, 56.7%, 37.4% at 200 ${\mu}g/m{\ell}$, respectively. Also, those extracts showed protective effect of DNA damage against the oxidative stress. Conclusion : These results indicated that the leaf extracts of Abeliophylli distichi Folium can be used as a natural antioxidants, which effectively inhibits the oxidative DNA damage.

XPS STUDY ON THE EFFECT OF LOW-ENERGY ELECTRON IRRADIATION ON DNA DAMAGE BY Fe3+ ION

  • Noh, Hyung-Ah;Park, Yeun-Soo;Cho, Hyuck
    • Journal of Radiation Protection and Research
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    • v.40 no.2
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    • pp.87-91
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    • 2015
  • We have employed X-ray photoelectron spectroscopy (XPS) technique to examine the combined effects of low-energy electron (LEE) irradiation and $Fe^{3+}$ ion on DNA damage. pBR322 plasmid DNA extracted from E. coli ER2420 was used for preparing DNA-$Fe^{3+}$ sample. The C1s XPS spectra were scanned for LEE-irradiated and LEE-unirradiated samples and then curve-fitted. For the samples with LEE irradiation only or with Fe ion only, no significant changes from pure DNA samples were observed - a single effect of either $Fe^{3+}$ ion or LEE irradiation did not cause a significant damage. However, when these two components were combined, the DNA damage was increased quite significantly, compared to the sum of DNA damages caused by $Fe^{3+}$ ion and by LEE irradiation independently. This observation is consistent with our previous results [Radiat. Res. 177, 775 (2012)] which was done using gel-electrophoresis technique. Partial interpretation of the observed spectrum peaks was also attempted.

Plasmid DNA damage by neutron and ${\gamma}$-ray in the presence of BSH (BSH 존재시 중성자 및 ${\gamma}$-ray 조사에 따른 plasmid DNA의 손상)

  • Chun, Ki-Jung;Seo, Won-Sook
    • Journal of Radiation Protection and Research
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    • v.31 no.2
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    • pp.65-68
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    • 2006
  • In this study, the extent of plasmid DNA damage was observed according to concentration of BSH(Boron Sulfhydryl Hydride) and irradiation doses of neutron and ${\gamma}$-ray. The plasmid used was both pBR 322 (2870 bp) and ${\Phi}X174$ RF(5386 bp) DNA. Plasmid DNA damage by irradiation in the presence of BSH was analyzed by agarose gel electrophoresis. In the neutron experiment, DNA damage of both plasmid DNAs was increased according to increasing the concentration of BSH and neutron doses. But in the ${\gamma}$-ray experiment, there appeared no dose dependency as compared to the neutron experiment. The extent of the plasmid DNA damage in the presence of BSH was somewhat different according to irradiation by neutron or ${\gamma}$-ray.

The Effect of Carnosine Extracted from Eels Anguilla japonica on Oxidative DNA Damage Induced by Hydrogen Peroxide and the DNA Repair Capacity of Human Leukocytes (뱀장어(Anguilla japonica) 추출 Carnosine이 과산화수소로 유도된 인체 백혈구의 DNA 손상과 Repair에 미치는 효과)

  • Song, Ho-Su
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.5
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    • pp.520-526
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    • 2017
  • Carnosine was recently reported to protect against the DNA damage induced by oxidative stress. In this study, we investigated the protective effect of eel Anguilla japonica carnosine extracts prepared using different methods (heat treatment extracts, HTEs; ion exchange chromatography, IEC; ultrafiltration permeation, UFP) on leukocyte DNA damage using the comet assay. Human leukocytes were incubated with extracts of eel carnosine at concentrations (of 10, 50, $100{\mu}g/mL$), and then subjected to an oxidative stimulus [$200{\mu}M$ hydrogen peroxide ($H_2O_2$)]. Pretreatment of the cells for 30 min with carnosine significantly reduced the genotoxicity of $H_2O_2$ measured as DNA strand breaks. The protective effects of the three types of extract (HTE, IEC, and UFP) increased with concentration. At the highest concentration (100 g/mL). there were no statistical differences in oxidative damage between each extract treatment and PBS-treated negative controls. When leukocytes were incubated with carnosine for 30 min after exposure to $H_2O_2$. the protective ability of each extract changed. Therefore, eel carnosine inhibits the $H_2O_2$ induced damage to cellular DNA in human leukocytes, supporting the protective effect of this compound against oxidative damage.

Comparison of the Protective Effect of Antioxidant Vitamins and Fruits or Vegetable Juices on DNA Damage in Human Lymphocyte Cells Using the Comet Assay (Comet Assay를 이용한 항산화 비타민과 과일.야채즙의 인체 임파구 세포 DNA 손상 감소 효과 비교)

  • 전은재;박유경;김정신;강명희
    • Journal of Nutrition and Health
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    • v.37 no.6
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    • pp.440-447
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    • 2004
  • In this study the in vitro protective effects of several antioxidant vitamins (vitamin C, $\alpha$-tocopherol, $\beta$-carotene), fruits and vegetables (strawberry, tangerine, orange and 100% orange juice, carrot juice), on the levels of isolated human lymphocyte DNA damage was measured using Comet assay. Comet assay has been used widely to assess the level of the DNA damage in the individual cells. Lymphocytes were pre-treated for 30 minutes with antioxidant vitamins (10, 50, 100, 500 $\mu$M) or fruits$.$vegetables (10, 100, 500, 1000 $\mu$g/ml), an4 then oxidatively challenged with 100 $\mu$M $H_2O$$_2$ for 5 min at 4$^{\circ}C$. The protective effect of antioxidant vitamins against DNA damage at a concentration of 50 $\mu$M were 50% in vitamin C, 32% in $\alpha$-tocopherol, whereas, fJ-carotene showed a 55% protection at a dose as low as 10 $\mu$M. The inhibitory effects of DNA damage by strawberry, tangerine, orange, orange juices, carrot juices were 50 - 60% with wide ranges of doses. The results of the present study indicate that most the antioxidant vitamins and fruits$.$vegetables juices produced a significant reduction in oxidative DNA damage.