• Title/Summary/Keyword: DNA Damage

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Nuclear DNA Damage and Repair in Normal Ovarian Cells Caused by Epothilone B

  • Rogalska, Aneta;Marczak, Agnieszka
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.15
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    • pp.6535-6539
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    • 2015
  • This study was designed to assess, whether a new chemotherapeutic microtubule inhibitor, Epothilone B (EpoB, Patupilone), can induce DNA damage in normal ovarian cells (MM14.Ov), and to evaluate if such damage could be repaired. The changes were compared with the effect of paclitaxel (PTX) commonly employed in the clinic. The alkaline comet assay technique and TUNEL assay were used. The kinetics of DNA damage formation and the level of apoptotic cells were determined after treatment with IC50 concentrations of EpoB and PTX. It was observed that PTX generated significantly higher apoptotic and genotoxic changes than EpoB. The peak was observed after 48 h of treatment when the DNA damage had a maximal level. The DNA damage induced by both tested drugs was almost completely repaired. As EpoB in normal cells causes less damage to DNA it might be a promising anticancer drug with potential for the treatment of ovarian tumors.

Protection of Radiation-Induced DNA Damage by Functional Cosmeceutical Poly-Gamma-Glutamate

  • Oh, Yu-Jin;Kwak, Mi-Sun;Sung, Moon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.28 no.4
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    • pp.527-533
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    • 2018
  • This study compared the radioprotective effects of high-molecular-weight poly-gamma-glutamate (${\gamma}-PGA$, average molecular mass 3,000 kDa) and a reduced form of glutathione (GSH, a known radioprotector) on calf thymus DNA damage. The radiation-induced DNA damage was measured on the basis of the decreased fluorescence intensity after binding the DNA with ethidium bromide. All the experiments used $^{60}Co$ gamma radiation at 1,252 Gy, representing 50% DNA damage. When increasing the concentration of ${\gamma}-PGA$ from 0.33 to $1.65{\mu}M$, the DNA protection from radiation-induced damage also increased, with a maximum of 87% protection. Meanwhile, the maximal DNA protection when increasing the concentration of GSH was only 70%. Therefore, ${\gamma}-PGA$ exhibited significant radioprotective effects against gamma irradiation.

Radiation Protective Effect of vitamin C and Cysteine on DNA Damage in Mice Splenic Lymphocytes by Single Cell Gel Electrophoresis Assay (단세포 겔 전기영동법을 이용한 생쥐 비장 림프구 DNA 손상에 대한 비타민 C 및 시스테인의 방사선 방어효과)

  • 천기정;김진규;김봉희
    • Environmental Analysis Health and Toxicology
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    • v.16 no.1
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    • pp.17-20
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    • 2001
  • The alkaline comet assay, employing a single-cell gel electrophoresis(SCGE), is a rapid, simple and sensitive technique for visualizing and measuring DNA damage leading to strand breakage in individual mammalian cells. The protecting effect of pretreatment with vitamin C and cysteine on the DNA damage of gamma ray was investigated in mice splenic lymphocytes. Vitamin C and cysteine were administered orally for five consecutive days before irradiation. Four week old ICR male mice were irradiated wish 3.5Gy of γ-radiation and were sacrificed 3 days later. Spleens were taken for DNA damage examination by Comet assay and the tail moments of DNA single -strand breaks in tole splenic lymphocytes were evaluated. The results show that pretreatment with vitamin C and cysteine were effective in protecting against DNA damage by gamma ray. Administration of antioxidants like vitamin C and cysteine to mice before irradiation was effective in reducing the tail moment of splenic lymphocytes DNA.

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Application of the Alkaline Comet Assay for Detecting Oxidative DNA Damage in Human Biomonitoring (인체 산화적 DNA손상에 대한 Human Biomonitoring도구로서 Alkaline Comet Assay의 활용 가능성 연구)

  • 박은주;강명희
    • Journal of Nutrition and Health
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    • v.35 no.2
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    • pp.213-222
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    • 2002
  • The alkaline comet assay has been used with increasing popularity to investigate the level of DNA damage in biomonitoring studies within the last decade in Western countries. The purpose of this study was to evaluate the usefulness of the alkaline comet assay as a biomarker of oxidative DNA damage for monitoring in the Korean population, and also to evaluate the effect of nutritional status and lifestyle factors on H2O2 induced oxidative DNA damage measured by the alkaline comet assay in human lymphocytes. The study population consisted of 61 healthy Korean male volunteers, aged 20-28. Epidemiological background data including dietary habits, smoking habits and anthropometrical measurements were collected through personal interviews. After blood collection, the comet assay in peripheral lymphocytes and plasma lipids analysis was carried out and the results analyzed. Tail moment (TM) and tail length (TL) of the comet assay were use\ulcorner to measure DNA damage in the lymphocytes of the subjects. Statistically significant (p < 0.05) positive correlations were observed between DNA damage (TM or TL) and smoking habits expressed as cigarettes smoked per day and pack years (r = 0.311 and 0.382 for TM, r = 0.294 and 0.350 for TL, respectively). There were also significant positive correlations between DNA damage parameter and waist-hip ratio. Higher plasma triglyceride levels were associated with increased damage to DNA. There were no correlations between the consumption frequencies of vegetables and DNA damage to the subjects. However, consumption frequencies of fruit and fruit juice intake were inversely associated with the TM and TL. The results indicate that die comet assay is a simple, rapid and sensitive method for detecting lymphocyte DNA damage induced by cigarette smoking. Consumption of fruit or fruit juices could potentiall modify the damaged DNA in the human peripheral lymphocytes of young Korean men.

Increased DNA Damage of Lymphocytes in Korean Male Smokers (일부 한국 성인 남성 흡연자들의 림프구 DNA 손상의 증가)

  • Lee, Joo-Hyun;Oh, Eun-Ha;Lee, June-Young;Sul, Dong-Geun;Kim, Joo-Ja;Lee, Eun-Il
    • Journal of Preventive Medicine and Public Health
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    • v.40 no.1
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    • pp.16-22
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    • 2007
  • Objective : The purpose of this study was to evaluate the levels of DNA damage in human lymphocytes caused by smoking and other lifestyle factors. Methods : The study population consisted of 173 normal healthy male adults from 21 to 59 years old. The demographic and lifestyle variables were obtained from administered questionnaires. The level of lymphocytic DNA damage in the peripheral blood was evaluated by the Comet assay. Statistical analyses were done by general linear model analysis and Dunnett's multiple comparison. Results : The difference in DNA damage between smokers and non-smokers was statistically significant. The means for the Tail%DNA were found to be 10.48 in the current smokers and 9.60 in the non-smokers (p < 0.05). The tail moment means were 1.58 and 1.45 (p < 0.05) for the current smokers and non-smokers, respectively. The number of cigarettes smoked per day did not result in a significant difference in the level of DNA damage among the smokers. Other lifestyle factors such as age, and drinking and exercise habits were not related to DNA damage. Conclusions : The DNA damage in the lymphocytes of smokers was found to be significantly higher than that for non-smokers. However, the number of cigarettes smoked per day was not related to DNA damage. Further study is needed to evaluate the relationship between the amount of smoking and level of damage to DNA. In addition, the status of DNA repair activities should be assessed.

The DNA Damage by Linoleic Acid Hydroperoxide (Linoleic acid과산화물의 DNA 손상작용)

  • KIM Seon-Bong;KANG Jin-Hoon;BYUN Han-Seok;KIM In-Soo;PARK Yeung-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.20 no.6
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    • pp.569-572
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    • 1987
  • The DNA damage by linoleic acid hydroperoxide (LHPO) was investigated in a DNA-LHPO system at $37^{\circ}C$ to elucidate the DNA damage mechanism by lipid peroxidation products. LHPO shelved a great DNA damage with the increase of its concentrations. DNA was completely damaged in a LHPO-DNA(weight ratio, 2:3) system after incubation for 2 days. The degree of DNA ,damage by LHPO was greated than that of linoleic acid. In the quantitative analysis of DNA damage, the decreasing ratio of DNA content was $60\%$ in $84{\mu}g$ LHPO system incubated for 1 day compared to the control solution marked $30\%$. There were no participation of active oxygens on the DNA damage by LHPO.

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Suppressive Effects of Various Antioxidants on Melamine-induced Oxidative DNA Damage in Human Lymphocytes

  • Park, Seul-Ki;Lee, Mi-Young
    • Molecular & Cellular Toxicology
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    • v.5 no.3
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    • pp.243-249
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    • 2009
  • Melamine, which is used to produce melamine resin for various industrial applications, has a high nitrogen content by mass. For this reason, it has been illegally added to foods to increase their apparent protein content. In the present investigation, melamine-induced oxidative damage of human lymphocyte DNA was evaluated by Comet assay. The in vitro oxidative DNA damage caused by melamine increased in a dose-dependent manner. This DNA damage was significantly inhibited by treatment with ascorbate. Moreover, the traditional Korean medicinal herb, named Acanthopanax, red ginseng and green tea markedly reduced the DNA damage. Various edible plant extracts also inhibited melamine-induced oxidative DNA damage in vitro. Melamine enhanced intracellular ROS generation, and this effect was suppressed by treatment with various antioxidants.

The Effects of Purple Grape Juice Supplementation on Improvement of Antioxidant Status and Lymphocyte DNA Damage in Korean Smokers (포도주스의 보충섭취가 흡연성인의 혈장 항산화 영양상태 및 DNA 손상 개선에 미치는 영향)

  • 박은주;김정신;전은재;김혜영;박유경;강명희
    • Journal of Nutrition and Health
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    • v.37 no.4
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    • pp.281-290
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    • 2004
  • The purpose of this project was to evaluate whether daily fruit juice consumption could reduce the DNA damage in healthy subjects. The study was performed using 67 healthy volunteers (29 smokers, 38 nonsmokers) who were supple-mented with 480 m1 of grape juice for 8 weeks. Eight weeks of grape juice consumption did not change any anthropometric parameters. Lymphocyte DNA damage before the study was significantly greater (p<0.05) in smoker than nonsmoker, but, grape juice consumption significantly reduced DNA damage in both smoker (26%) and nonsmoker (I7%) to the level where there was no difference remained between the two groups after the intervention trial. This preventive effect of grape juice against DNA damage was not affected by sex of the subjects in non-smokers. Plasma $\alpha$-carotene, Iyco-pene and ${\gamma}$-totopherol was significantly increased after the trial in smokers, while erythrocyte catalase was significan-tly increased in both smokers and nonsmokers. Total radical-trapping antioxidant potential (TRAP) level in all subjects was significantly reduced after the intervention, while GSH-Px activity was increased only in nonsmokers. These results suggests that daily consumption of grape juice may protect DNA damage in peripheral lymphocytes, and supports the hypothesis that grape juice might exert their effect partially via a decrease in oxidative damage to DNA in humans partly by improving their antioxidative defense system.

Inactivation of Mad2B Enhances Apoptosis in Human Cervical Cancer Cell Line upon Cisplatin-Induced DNA Damage

  • Ju Hwan Kim;Hak Rim Kim;Rajnikant Patel
    • Biomolecules & Therapeutics
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    • v.31 no.3
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    • pp.340-349
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    • 2023
  • Mad2B (Mad2L2), the human homolog of the yeast Rev7 protein, is a regulatory subunit of DNA polymerase ζ that shares sequence similarity with the mitotic checkpoint protein Mad2A. Previous studies on Mad2B have concluded that it is a mitotic checkpoint protein that functions by inhibiting the anaphase-promoting complex/cyclosome (APC/C). Here, we demonstrate that Mad2B is activated in response to cisplatin-induced DNA damage. Mad2B co-localizes at nuclear foci with DNA damage markers, such as proliferating cell nuclear antigen and gamma histone H2AX (γ-H2AX), following cisplatin-induced DNA damage. However, unlike Mad2A, the binding of Mad2B to Cdc20 does not inhibit the activity of APC/C in vitro. In contrast to Mad2A, Mad2B does not localize to kinetochores or binds to Cdc20 in spindle assembly checkpoint-activated cells. Loss of the Mad2B protein leads to damaged nuclei following cisplatin-induced DNA damage. Mad2B/Rev7 depletion causes the accumulation of damaged nuclei, thereby accelerating apoptosis in human cancer cells in response to cisplatin-induced DNA damage. Therefore, our results suggest that Mad2B may be a critical modulator of DNA damage response.

Inhibition Effects of Persicaria amphibia (L.) Delarbre on Oxidative DNA Damage via ATM/Chk2/p53 pathway

  • So-Yeon Han;Hye-Jeong Park;Jeong-Yong Park;Seo-Hyun Yun;Mi-Ji Noh;Soo-Yeon Kim;Tae-Won Jang;Jae-Ho Park
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2021.04a
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    • pp.52-52
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    • 2021
  • Persicaria amphibia as an England native plant, is a rhizomatous perennial, one of the rather amphibious plants. Its aquatic form contains water-soluble sugars, starch, and protein. P. amphibia have up to 18% tannins in stems and rhizomes. Previous studies have confirmed the anti-inflammatory activity of live bacteria roots, but no studies on bioactivity are known. DNA damage responses (DDRs) pathways are considered a crucial factor affecting the alleviation of cellular damage. The ataxia-telangiectasia mutated and Rad3 related (ATM) and checkpoint kinase 2 (Chk2) pathways are the main pathways of DNA damage response. Also, p53 is a key integrator of cellular response to oxidative DNA damage, contributing repair, or leading transcription including apoptosis. In the present study, we conducted an investigation into the inhibitory effects of P. amphibia on oxidative DNA damage for confirming potential to complementary medicine and therapies. In conclusion, P. amphibia can provide protective effects against double-stranded DNA break (DSB) caused by oxidative DNA damage.

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