• Title/Summary/Keyword: tail length

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Changes of DNA Fragmentation by Irradiation Doses and Storage in Gamma-Irradiated Fruits (감마선 조사 과일류에서 조사선량과 저장기간에 따른 DNA Fragmentation의 변화)

  • Kim, Sang-Mi;Park, Eun-Ju;Yang, Jae-Seung;Kang, Myung-Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.4
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    • pp.594-598
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    • 2002
  • The changes in DNA damage were investigated during storage after irradiation. Kiwi, orange and pear were irradiated at 0.1, 0.3, 0.5, 0.7 and 1.0 kGy and stored for 3 months at 4$^{\circ}C$. The comet assay was applied to the sample seeds alt the beginning of irradiation and at the end of storage. Seeds were isolated and crushed, and the suspended cells were embedded in an agarose layer. After lysis of the cells, they were electrophoresed for 2 min and then stained. DNA fragmentation in seeds caused by irradiation was quantified as tail length and tail moment (tail length $\times$ % DNA in tail) by comet image analyzing system. Immediately after irradiation, the differences in tail length between unirradiated and irradiated fruit seeds were significant (p<0.05) in kiwi, orange and pear seeds. With in-creasing the irradiation doses, statistically significant longer extension of the DNA from the nucleus toward anode was observed. The results represented as tail moment showed similar tendency to those of tail length, but tile latter parameter was more sensitive than the former. Similarly even 3 months after irradiation, all the irradiated fruit seeds significantly showed longer tail length than the unirradiated controls. These results indicate that the comet assay could be one of the simple methods of detecting irradiated fruit seeds. Moreover, the method could detect DNA damage even after 3 months after irradiation.

Changes of DNA Fragmentation by Irradiation Doses and Storage in Gamma-irradiated Potato, Garlic and Ginger (감마선 조사된 감자, 마늘, 생강에서 조사선량과 저장기간에 따른 DNA fragmentation의 변화)

  • Lee, Hye-Jin;Park, Yoo-Kyoung;Yang, Jae-Seung;Kang, Myung-Hee
    • Journal of the Korean Society of Food Culture
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    • v.19 no.3
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    • pp.251-258
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    • 2004
  • The changes in DNA damage were investigated during storage after irradiation. Potato, garlic were irradiated at 0.05, 0.07, 0.1 and 0.15 kGy and stored for 3 months. Ginger was irradiated at 0.01, 0.02, 0.03, 0.04 and 0.05 kGy and stored for 1 month. The comet assay was applied to the sample immediately after irradiation and at the end of storage. Samples were isolated, grounded and the suspended cells were embedded in an agarose layer. After lysis of the cells, they were electrophoresed for 1 min. and then stained. DNA fragmentation in seeds caused by irradiation was quantified as tail length and tail moment (tail length ${\times}%$ DNA in tail) by comet image analyzing system. Right after irradiation, the differences in tail length between unirradiated and irradiated samples were significant(p<0.05) in potato, garlic and ginger. With increasing the irradiation doses, statistically significant longer extension of the DNA from the nucleus toward anode was observed. The results represented as tail moment showed similar tendency to those of tail length. Similarly in the stored samples, even 1 or 3 months after irradiation, all the irradiated samples significantly showed longer tail length than the unirradiated controls. These results indicate that the comet assay could be one of the simple methods of detecting irradiated samples. Moreover, the method could detect DNA damage even after 1 or 3 months after irradiation.

Changes of DNA fragmentation by Irradiation Doses and Storage in Gamma-irradiated Meats and Poultry (감마선 조사 육류, 가금류에서 저장전과 후의 조사선량에 따른 DNA fragmentation의 변화)

  • Lee, Hye-Jin;Kim, Sang-Mi;Park, Yoo-Kyoung;Yang, Jae-Seung;Kang, Myung-Hee
    • Journal of the Korean Society of Food Culture
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    • v.19 no.2
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    • pp.129-138
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    • 2004
  • The changes in DNA damage were investigated during storage after irradiation. Beef, pork and chicken were irradiated at 1.0, 3.0 and 5.0 kGy and stored for 6 months at $-20^{\circ}C$. The comet assay was applied to the sample muscles at the beginning of irradiation and at the end of storage. Muscles were isolated, sliced, and the suspended cells were embedded in an agarose layer. After lysis of the cells, they were electrophoresed for 2 min. and then stained. DNA fragmentation in tissues caused by irradiation was quantified as tail length and tail moment (tail length ${\times}$ % DNA in tail) by comet image analyzing system. Right after irradiation, the differences in tail length between unirradiated and irradiated muscles were significant(p<0.05) in beef, pork and chicken. With increasing the increasing doses, statistically significant longer extension of the DNA from the nucleus toward anode was observed. Similarly even 6 months after irradiation, all the irradiated muscles significantly showed longer tail length than the unirradiated controls. The results represented as tail moment showed similar tendency to those of tail length, but the latter parameter was more sensitive than the former. These results indicate that the comet assay could be one of the simple methods of detecting irradiated muscles. Moreover, this method suggest that using comet assay, we were able to detect DNA damage differences even after 6 months after irradiation.

POWER TAIL ASYMPTOTIC RESULTS OF A DISCRETE TIME QUEUE WITH LONG RANGE DEPENDENT INPUT

  • Hwang, Gang-Uk;Sohraby, Khosrow
    • Journal of the Korean Mathematical Society
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    • v.40 no.1
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    • pp.87-107
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    • 2003
  • In this paper, we consider a discrete time queueing system fed by a superposition of an ON and OFF source with heavy tail ON periods and geometric OFF periods and a D-BMAP (Discrete Batch Markovian Arrival Process). We study the tail behavior of the queue length distribution and both infinite and finite buffer systems are considered. In the infinite buffer case, we show that the asymptotic tail behavior of the queue length of the system is equivalent to that of the same queueing system with the D-BMAP being replaced by a batch renewal process. In the finite buffer case (of buffer size K), we derive upper and lower bounds of the asymptotic behavior of the loss probability as $K\;\longrightarrow\;\infty$.

Detection of Irradiated Grains Using the DNA ‘Comet Assay’ (DNA ‘Comet Assay’를 이용한 곡류의 방사선 조사 여부 확인)

  • Kim, Choong-Ki;Yang, Jae-Seung;Lee, Hae-Jung
    • Korean Journal of Food Science and Technology
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    • v.31 no.4
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    • pp.906-911
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    • 1999
  • This study was to determine whether DNA 'Comet Assay' can be applied to the detection of grains irradiated with low doses of Co-60 gamma radiation. Sesame, perilla, wheat, barley and rice were exposed to different doses of 0.1, 0.3, 0.5, 0.7 and 1.0 kGy. The cells isolated from the samples were embedded in a agarose gel on a microscope slide, lysed in lysis solution, and subjected to electrophoresis. DNA and its fragments migrated in the gel produced the characteristic pattern of DNA comet, of which the tail length was measured in a microscope. All the samples irradiated at 0.3 kGy and higher were applicable to detect post-irradiation by the tail length of their comets. Irradiated samples showed comets with long tails and their tail length increased with the dose, while unirradiated samples showed no or very short tails. Especially, sesame, perilla and wheat irradiated at 0.1 kGy could be distinguished from unirradiated samples by visual inspection of the slide in a microscope. Thus, DNA 'Comet Assay' might be applied to the detection of irradiated grains as a simple, inexpensive and rapid screening test.

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Exploitation of the Dose/Time-Response Relationship for a New Measure of DNA Repari in the Single-Cell Gel Electrophoresis (Comet) Assay

  • Kim, Byung-Soo;Edler, Lutz;Park, Jin-Joo;Fournier, Dietrich Von;Haase, Wulf;Sautter-Bihl, Mare-Luise;Hagmuller, Egbert;Gotzes, Florian;Thielmann, Heinz Walter
    • Toxicological Research
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    • v.20 no.2
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    • pp.89-100
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    • 2004
  • The comet assay (also called the single-cell gel electrophoresis assay) has been widely used for detecting DNA damage and repair in individual cells. Since the conventional methods of evaluating comet assay data using frequency statistics are unsatisfactory we developed a new quantitative measure of DNA damage/repair that is based on all information residing in the dose/time-response curves of a comet experiment. Blood samples were taken from 25 breast cancer patients before undergoing radiotherapy. The comet assay was performed under alkaline conditions using isolated lymphocytes. Tail DNA, tail length, tail moment and tail inertia of the comet were measured for each patient at four doses of $\gamma$-rays (0, 2, 4 and 8 Gy) and at four time points after irradiation (0, 10, 20 and 30 min) using 100 cells each. The resulting three-dimensional dose-time response surface was modeled by multiple regression, and the second derivative, termed 2D, on dose and time was determined. A software module was programmed in SAS/AF to compute 2D values. We applied the new method successfully to data obtained from cancer patients to be assessed for their radiation sensitivity. We computed the 2D values for the four damage measures, i.e., tail moment, tail length, tail DNA and tail inertia, and examined the pairwise correlation coefficients of 2D both on the log scale and the unlogged scale. 2D values based on tail moment and tail DNA showed a high correlation and, therefore, these two damage measures can be used interchangeably as far as DNA repair is concerned. 2D values based on tail inertia have a correlation profile different from the other 2D values which may reflect different facets of DNA damage/repair. Using the dose-time response surface, other statistical models, e.g., the proportional hazards model, become applicable for data analysis. The 2D approach can be applied to all DNA repair measures, Le., tail moment, tail length, tail DNA and tail inertia, and appears to be superior to conventional evaluation methods as it integrates all data of the dose/time-response curves of a comet assay.

Discrimination of Irradiated Beef Using Comet Assay (Comet assay를 이용한 방사선 조사육의 판별)

  • Jeong, Seok-Kyu;Park, Jong-Heum;Ji, Seung-Taek;Park, Kum-Ju;Kim, Hai-Hong;Hyun, Chang-Kee;Shin, Heuyn-Kil
    • 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.

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Gender-Specific Changes of Plasma MDA, SOD, and Lymphocyte DNA Damage during High Intensity Exercise (고강도 운동 시 성별에 따른 혈장 MDA, SOD 및 임파구 DNA 손상 변화)

  • Cho, Su-Youn;Chung, Young-Soo;Kwak, Yi-Sub;Roh, Hee-Tae
    • Journal of Life Science
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    • v.21 no.6
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    • pp.838-844
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    • 2011
  • The purpose of this study was to investigate gender-specific changes of plasma MDA, SOD, and lymphocyte DNA damage during high intensity exercise. In this study, 17 healthy male and 18 healthy female college students ran on a treadmill at 85%$VO_{2max}$ until the point of all-out. Blood-collecting was carried out five times (Rest, Ex-Exha, R0.5h, R4h and R24h), and with the collected blood, plasma malondialdehyde (MDA), superoxide dismutase (SOD), and lymphocyte DNA damage were analyzed. Plasma MDA and SOD concentration increased significantly at the Ex-Exha (p<0.05), and there were no significant differences in gender. For the degree of lymphocyte DNA damage, all %DNA in the tail, tail length and tail moment increased significantly at the Ex-Exha (p<0.05), and %DNA in the tail and tail length were significantly higher in the male group than in the female group (p<0.05). These results suggest that acute high intensity exercise not only causes oxidative stress but also brings about lymphocyte DNA damage. In addition, it was found that males showed higher DNA damage than females in terms of oxidative stress subject to high intensity exercise. Nevertheless, further subsequent studies are required in order to better understand the mechanism behind DNA damage varying with gender, in a way that takes into consideration physical fitness, hormonal level, exercise intensity and duration - additional factors which might affect DNA damage.

Effect of Deer Antler Drink Supplementation on Blood Pressure, Blood Glucose and Lymphocyte DNA Damage in Type 2 Diabetic Patients (당뇨환자에 있어서 녹용추출물의 섭취가 혈압, 혈당 및 임파구 DNA 손상에 미치는 영향)

  • Kim Hye-Young;Jeon Eun-Jae;Park Yoo Kyoung;Kang Myung-Hee
    • Journal of Nutrition and Health
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    • v.37 no.9
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    • pp.794-800
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    • 2004
  • Deer Antler has been known for its traditional oriental medicinal properties and has been widely used to promote growth, boost immune function, treat blood loss and chronic joint pain. Recent study showed imported (New Zealand) Deer Antler was beneficial in reducing the side effects of cancer treatments. However, there was no intervention study conducted on the effect of Korean Deer Antler on reducing the oxidative stress to patients with diabetes. One of the sensitive ways to measure endogenous oxidative stress is by measuring cellular DNA damage using single cell gel electrophoresis (COMET assay). This study was conducted to investigate the possible beneficial effect of commercial Deer Antler drink (provided by Chung-yang Deer Farm) on lymphocyte DNA damage and blood glucose of diabetic patients. Ten patients (4 men, 6 women) participated in the study and consumed 2 pouches of Deer Antler drink every day for 20 days. Blood was collected on the morning before and after the intervention for lymphocyte isolation and blood glucose analysis. Both systolic and diastolic blood pressure showed a tendency to decrease but did not reach statistical significance after the trial. Blood glucose level was not affected by the supplementation. After the intervention, over 50% reduction were noted in the cellular DNA damage, expressed as tail length (TL) and tail moment (TM: tail length ${\times}$ percent tail DNA) . Although we did not obtain beneficial effect on lowering blood glucose levels in the patients, this results suggest that Deer Antler may initially act in protecting endogenous DNA damage in short-term experiment.

DNA damages with Fpg/Endo Ⅲ FLARE Assay in cynomolgus monkeys exposed to stainless steel welding fume (용접흄 흡입노출 영장류에서 Fpg/Endo Ⅲ FLARE Assay를 이용한 DNA 손상 및 회복)

  • Rim, Kyung Taek;Kim, Soo Jin;Chung, Yong Hyun;Kim, Hyeon Yeong;Maeng, Seung Hee;Yu, Il Je
    • 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.