• Journal of radiological science and technology
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• v.34 no.4
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• pp.333-339
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• 2011

The purpose of this study is to compare DNA repair characteristics of normal fibroblast cell (MRC-5) and neuroblastoma cell (SK-N-SH) induced by proton beam. Cells were irradiated with 2Gy, 5Gy and 8Gy proton beam. The rate of DNA rejoining was measured by alkaline version of the comet assay. After a repair time, tail moment was measured again. The tail moment of MRC-5 was lower than SK-N-SH. However, after 8Gy of exposure, the tail moment of MRC-5 was measured as 50.320223.17155 which represents dangerous level of DNA damage. The cells were repaired practically within 25 hours after 2 and 5Gy of exposure while they were not fully recovered after 8Gy of exposure. Especially, tail moment of MRC-5 after 25 hours was 18.15364.42849. In the distal declining edge of SOBP, the RBE value is increased by high LET. The RBE differences of SOBP in high-dose were greater than low-dose. After the high-dose exposure, MRC-5 of normal fibroblast cell could lead to lasting DNA damage as shown in this study. In conclusion, we has to pay special attention when the region of the treatment volume is close to sensitive tissues.

• 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.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.6
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• pp.1025-1029
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• 2000

This study was conducted to investigate whether a DNA comet assay could be applied for identifying irradiated pork and beef. Pork and beef were irradiated with Co-60 gamma rays at 0.1, 0.3, 0.5, 0.7 and 1.0 kGy, and stored in a freezer Cells separated from the samples were embedded in agarose gel on a slide, dissolved in a lysis solution, and electrophoresed at 2 V/cm for 2.0 min by horizontal electrophoesis. The cells were then stained with a silver staining in order to visualize the DNA using a micro-scope. The DNA fragments of the irradiated cells stretched or migrated out of the cells and formed tails towards the anode, giving the appearance of comets, while unirradiated cells formed very short or no tails. The distance of DNA migration increased with irradiation dose. Since the statistical analysis showed a significant correlation between tail length and irradiation dose, a DNA comet assay could provide not only identification but also estimation of the irradiation dose for irradiated beef and pork.

• Journal of Nutrition and Health
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• v.43 no.6
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• pp.570-577
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• 2010

Water gets magnetically charged when it is contacted with a magnet. Although magnetic water products have been promoted since the 1930's, they have received very little recognition due to questionable effectiveness. Diethylnitrosamine (DEN) is a widely occurring nitrosamine that is one of the most important environmental carcinogens primarily inducing tumors of liver. In this study, the effect of magnetized water supplementation on lymphocyte DNA damage in ICR mice treated with DEN was evaluated using the Comet assay. Mice were divided into 3 groups: control, DEN, and DEN + magnetized water group. Fifteen mice were maintained in each group for the entire experimental period of 6, 12 and 18 weeks. Five mice in each group were sacrificed at 6, 12, and 18th weeks, followed by the Comet assay using the blood obtained from heart puncture of the mice. The level of lymphocyte DNA damage reflected by tail moment and other DNA damage indices of tail DNA (%) or tail length of the magnetized water group were significantly decreased after the 6th, 12th and 18th weeks of supplementation compared with the positive control, the DEN group. The relative DNA damage of the magnetized water groups compared to the DEN control group after 6th, 12th, and 18th weeks of supplementation were 42.2%, 40.8%, and 32.9% for DNA in tail, 31.2%, 32.6%, and 21.3% for tail length, and 33.8%, 33.8%, and 24.6% for tail moment, respectively. This is the first report demonstrating that magnetized water may be involved in the lowering effect of the DNA damage in DEN-treated ICR mice. This result suggests that the magnetized water might have minimized the DNA damage by improving the antioxidant status of the mice. However, further studies are needed to characterize the condition of the magnetization and examine the long-term effect of the water product.

• Korean Journal of Food Science and Technology
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• v.32 no.4
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• pp.747-754
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• 2000

DNA damages in post-mortem bovine muscle samples caused by gamma irradiation at doses of 1 to 10 kGy were determined by Comet assay. When the cell extract was prepared in a physical method and followed by neutral lysis and neutral electrophoresis, the optimal comet images could be obtained. DNA damages were evaluated from the mean tail length, the distributions of comet images in 4 groups divided by tail length and the relative damage index (RDI) values calculated from the distribution pattern. The mean tail length and RDI value were increased by increasing the irradiation dose, and the RDI value was found to be useful as an index for discriminating of irradiation and measuring the irradiated dose. Blind tests using Korean domestic (Hanwoo) and imported beef samples showed a higher RDI value for the latter. However, the value was lower than those of irradiated samples indicating that the cause of DNA damages in the imported beef samples might be not irradiation but low-temperature treatments. It was concluded from the results of this study that the irradiated beef and its irradiated dose could be detected and predicted by Comet assay.

• Journal of Preventive Medicine and Public Health
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• v.37 no.4
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• pp.373-380
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• 2004

Objectives : There has been gradually increasing concern about the adverse health effects of electromagnetic radiation originating from cell phones which are widely used in modern life. Cell phone radiation may affect human health by increasing free radicals of human blood cells. This study has been designed to identify DNA damage of blood cells by electromagnetic radiation caused by cell phone use. Methods : This study investigated the health effect of acute exposure to commercially available cell phones on certain parameters such as an indicator of DNA damage for 14 healthy adult volunteers. Each volunteer during the experiment talked over the cell phone with the keypad facing the right side of the face for 4 hours. The single cell gel electrophoresis assay (Comet assay), which is very sensitive in detecting the presence of DNA strand-breaks and alkali-labile damage in individual cells, was used to assess peripheral blood cells (T-cells, B-cells, granulocytes) from volunteers before and after exposure to cell phone radiation. The parameters of Comet assay measured were Olive Tail Moment and Tail DNA %. Results : The Olive Tail Moment of B-cells and granulocytes and Tail DNA % of B-cells and granulocytes were increased by a statistically significant extent after 4-hour use of a cell phone compared with controls. Conclusion : It is concluded that cell phone radiation caused the DNA damage during the 4 hours of experimental condition. Nonetheless, this study suggested that cell phone use may increase DNA damage by electromagnetic radiation and other contributing factors.

• Clinical and Experimental Reproductive Medicine
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• v.46 no.4
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• pp.147-151
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• 2019

Objective: The aim of this study was to investigate DNA fragmentation status in human spermatozoa according to specific tail swelling patterns determined via hypo-osmotic swelling test (HOST). Methods: Frozen semen samples from 21 healthy donors were thawed and prepared by the swim-up technique for use in intracytoplasmic sperm injection. The semen samples were treated for 5 minutes as part of the HOST procedure and then underwent the sperm chromatin dispersion test using a Halosperm kit. DNA fragmentation status (large halo, medium halo, small halo, no halo, or degraded) and the specific tail swelling pattern ("a"-"g") were assessed at the level of a single spermatozoon. A total of 42,000 spermatozoa were analyzed, and the percentage of spermatozoa without DNA fragmentation (as evidenced by a large or medium halo) was assessed according to the specific tail swelling patterns observed. Results: The HOST examinations showed that > 93% of spermatozoa across all types displayed no DNA fragmentation. The percentage of spermatozoa without DNA fragmentation was 100% in type "d", 98.67% in type "g", and 98.17% in type "f" spermatozoa. Conclusion: We found that the type "d" spermatozoa displayed no DNA fragmentation, but the other types of spermatozoa also displayed very low rates of DNA fragmentation. This result may be associated with the processing of the spermatozoa by density gradient centrifugation and the swim-up technique.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.17 no.4
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• pp.272-281
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• 2007

선박제조업을 비롯한 운송업 및 건축업 등의 다양한 분야에서 용접기술이 이용되어 옴에 따라 용접근로자들에 대한 산업보건학적 관심이 높아지고 있다. 노출정도가 다양하기는 하지만 용접흄은 6가 크롬을 비롯한 금속화합물과 유해가스, 화학물질 등을 복합적으로 포함하고 있는 스테인레스 스틸 용접흄에 대한 유전독성영향을 평가하기 위하여 흡입챔버를 이용, 실험동물인 영장류에 스테인레스 스틸 용접흄을 노출시키고 혈액 내 lymphocytes에 생성된 용접흄 노출농도 및 시간별 DNA 손상정도 및 그 회복효소를 측정함으로써, 유해성이 완전하게 확인되지 않은 용접흄에 노출되어 나타날 수 있는 암을 비롯한 심각한 건강영향을 예방하기 위한 각 지표들을 찾아 그 유용성을 비교하고자 하였다. 영장류를 노출시키기 위해 robotic arm을 장치한 영장류 흡입노출 시스템을 개발하였으며, 이 노출 시스템을 이용하여 수컷 영장류 6마리에 대해 용접흄 노출시험을 실시하였는데 실험군은 대조군 2, 저농도 ($31mg/m^3$) 노출군 2, 고농도 ($63mg/m^3$) 노출군 2마리로 구성하였고, 1일 2시간씩 일주일에 5일 동안 용접흄에 노출시켰다. 노출 농도는 지속적으로 모니터링 하였고, 노출과정 중에 영장류의 혈액을 채취하여 lymphocytes를 분리, 단세포 DNA 손상을 선별하기 위해 DNA 손상회복 효소인 E. coli formamidopyrimidine-DNA glycosylase (Fpg)와 endonuclease Ⅲ (Thymine Glycol-DNA glycosylase) 투여와 Comet asaay (single cell gel electrophoresis, 단세포겔전기영동기법)를 결합시켜 이용하는 Fpg/Endo III FLARE 분석법을 사용하였다. Fpg enzyme에 의한 olive tail moment값의 변화는 16주 노출군부터 노출부검(34주)군 까지 노출농도가 높아짐에 따른 olive tail moment 기하평균 값의 양 반응관계를 보기는 어렵지만, 고농도군의 경우 27주 노출군에서 가장 높은 olive tail moment 값을 보이고 이후 차츰 감소하였다. 한편 16주에서 22주까지의 노출기간에서는 대조군에 비해 노출군에서 DNA손상정도(olive tail moment값)는 모두 유의하게 높았으나, 6, 12, 18, 25, 31, 33, 35주간 노출하였을 때는 다른 결과를 보였다. 각 실험군의 Fpg enzyme에 의한 tail length값의 분포를 살펴볼 때, 저농도군 및 고농도군에서 27주간 노출하였을 때 가장 높은 tail length 값을 보이고 이후 차츰 감소하는 경향을 보였다. 또한 16, 22주간 노출하였을 때 대조군에 비해 노출군에서 tail length 값이 유의하게 높았으나, 20주간에서만 양 반응관계가 관찰되었고, 다른 주간에서는 양 반응 및 기간 반응관계를 나타내지는 않았다. Endo III enzyme에 의한 olive tail moment값의 변화는 기간별 노출군에서 대조군에 비해 높은 DNA손상정도(olive tail moment값)를 나타내는 결과들이 있었지만, 10, 12, 16, 22, 25, 31주간 노출하였을 때 등 상당수 노출기간에서 반응관계를 나타내지는 않았다. 각 실험군의 Endo III enzyme에 의한 tail length값의 분포를 살펴볼 때, 18, 20, 27, 33주간 노출하였을 때 대조군에 비해 노출군에서 tail length 값이 조금 높았지만, 양 반응 및 기간 반응관계를 보이지 않았고 수치의 크기가 불규칙하게 변화하였다. 즉, DNA에 있어 산화된 pyrimidine을 형성하여 손상된 부위의 염기를 제거함으로써 AP site (abasic site)를 만들고 이들이 Comet assay를 통해 break로 전환된 것을 포함한 DNA손상을 측정하기 위하여 endonuclease III (Endo III)를 첨가시킨 Endo III FLARE 분석법을 실시한 결과, 본 연구에서 나타난 결과는 용접흄 노출 영장류에서 Olive tail moment 및 tail length 공히 노출량 및 노출기간 반응관계를 볼 수 없었다. Endo III FLARE 분석법을 통한 산화적 DNA 손상지표는 영장류에 적용하기에는 적응반응현상으로 대조군과 유의한 차이도 관찰할 수 없었고 더욱이 역으로 대조군에서의 자연발생적 수치가 더 높아질 수 있어 용접흄 노출 영장류의 모니터링 지표로 사용하기에는 제한점이 있었다.

• Horticultural Science & Technology
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• v.33 no.2
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• pp.242-250
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• 2015

Insertional mutagenesis induced by T-DNA or transposon tagging offers possibilities for analysis of gene function. However, its potential remains limited unless good methods for detecting the target locus are developed. We describe a PCR technique for efficient identification of DNA sequences adjacent to the inserted T-DNA in a higher plant, Chinese cabbage (Brassica rapa ssp. pekinensis). This strategy, which we named variable argument thermal asymmetric interlaced PCR (VA-TAIL PCR), was designed by modifying a single-step annealing-extension PCR by including a touch-up PCR protocol and using long gene-specific primers. Amplification efficiency of this PCR program was significantly increased by employing an autosegment extension method and linked sequence strategy in nested long gene-specific primers. For this technique, arbitrary degenerate (AD) primers specific to B. rapa were designed by analyzing the Integr8 proteome database. These primers showed higher accuracy and utility in the identification of flanking DNA sequences from individual transgenic Chinese cabbages in a large T-DNA inserted population. The VA-TAIL PCR method described in this study allows the identification of DNA regions flanking known DNA fragments. This method has potential biotechnological applications, being highly suitable for identification of target genomic loci in insertional mutagenesis screens.

• 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.