• Korean Journal of Environmental Agriculture
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• v.34 no.4
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• pp.350-354
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• 2015

BACKGROUND: Botanical insecticides, especially Azadirachta Indica extract (AIE) and Sophorae radix extract (SRE) are widely used in Agriculture field. In our previous studies on genotoxicity test of AIE and SRE samples, a suspicious clastogenic properties was shown. Herein, we investigated the DNA damage effect of these botanical insecticide samples through the in vitro comet assay. METHODS AND RESULTS: Chinese hamster lung (CHL) fibroblast cell line was used, and methyl methanesulphonate was as positive control. Respective two samples of AIE and SRE were evaluated using Single Cell Gel Electrophoresis (Comet) assay and measured as the Olive tail moment (OTM). Results from this study indicated that all tested AIE and SRE samples did not show DNA damage in comet assay using CHL cells, compared with control. CONCLUSION: AIE and SRE samples used in this study were not cause genetic toxicity and are suitable for use as organic materials.

• Environmental Mutagens and Carcinogens
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• v.21 no.2
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• pp.89-94
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• 2001

Recently, single cell gel electrophoresis, also known as comet assay, is widely used for the detection and measurement of DNA strand breaks in vitro and in vivo in many toxicological fields such as radiation exposure, human monitoring and toxicity evaluation. As well defined, comet assay is a sensitive, rapid and visual method for the detection of DNA strand breaks in individual cells. Briefly, a small number of damaged cells suspended in a thin agarose gel on a microscope slide were lysed, unwinded, electrophoresed, and stained with a fluorescent DNA binding dye. The electric current pulled the charged DNA from the nucleus such that relaxed and broken DNA fragments migrated further. The resulting images which were subsequently named for their appearance as comets, were measured to determine the extent of DNA damages. However, some variations could be occurred in procedures, laboratories's conditions and kind of cells used. Hence, to overcome and to harmonize these matters in comet assay, International Workshop on Genotoxicity Test Procedure (IWGTP) was held with several topics including comet assay at Washington D.C. on March, 1999. In spite of some consensus in procedures and conditions in IWGTP, there are some problems still remained to be solved. In this respect, we attempted to set the practical optimal conditions in the experimental procedures such as lysis, unwinding, electrophoresis and neutralization conditions and so on. First of all, we determined optimal lysis and unwinding time by using 150 $\mu$M methyl methanesulfonate (MMS) which is usually used concentration. And then, we determined optimal positive control concentrations of benzo(a)pyrene (BaP) and MMS in the presence and absence of S9 metabolic activation system, respectively.

• Journal of Nutrition and Health
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• v.39 no.1
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• pp.18-27
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• 2006

Smoking is well known to be associated with increased indices of tree radical-mediated damage of DNA, indicating that smoking may exacerbate the initiation and propagation of oxidative stresses, which are potential underlying processes in the pathogenesis of many diseases. The purpose of this study was to evaluate whether a daily regimen of green vegetable drink supplementation to smokers can be protective against endogenous lymphocytic DNA damage and whether it could enhance other antioxidant status. Twenty nonsmokers and nineteen smokers aged 23-60 were given 240 ml of green vegetable drink every day for 8 weeks in addition to their normal diet, and blood samples were drawn before and after the intervention. The 8 weeks of green vegetable drink consumption resulted in a significant decrease (p = 0.000, by paired t-test) in lymphocyte DNA damage expressed by TL (before: $63.13{\pm}1.05$ vs after: $37.86{\pm}10.83$, before: $66.73{\pm}1.24$ vs after: $36.51{\pm}1.13$), TM (before: $14.55{\pm}0.61$ vs after: $6.61{\pm}0.25$, before: $15.36{\pm}0.45$ vs after: $6.65{\pm}0.38$) and $\%$ DNA in tail (before: $19.7{\pm}0.41$ vs after: $16.6{\pm}0.37$, before: $20.6{\pm}0.31$ vs after: $17.1{\pm}0.5$) in both nonsmokers and smokers respectively. Vitamin C and TRAP level was not significantly changed after the supplementation. In conclusion, these results support the hypothesis that green vegetable drink exert a cancer-protective effect partially via a decrease in oxidative damage to DNA.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2005.11a
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• pp.161-161
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• 2005

• Journal of Radiation Industry
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• v.8 no.2
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• pp.89-95
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• 2014

This study was conducted to determine the optimal dose of gamma-ray on the growth and nucleus DNA damage for mutation breeding in licorice. Gamma-rays irradiated to dry seeds with various doses (0 to 1000 Gy). Significant decreases in germination rate (%), survival rate (%) and growth characteristics (plant height, number of leaves, root length and fresh weight) were observed by dose of increased. $LD_{50}$ (lethal dose) was approximately 400 Gy to 500 Gy. Also, reduction doses ($RD_{50}$) of plant height, number of leaves, root length and flash weight were 428 Gy, 760 Gy, 363 Gy and 334 Gy, respectively. It is supplest that the optimal dose of gamma irradiation for licorice mutation induction might be about 400 Gy in this study. We also conducted comet assay to observe nucleus DNA damage due to gamma irradiation. In comet assay, a clear difference was identified over 300 Gy treatments. With increasing doses of gamma-ray in the range of 100 to 1000 Gy, the rate of head DNA was decreased significantly from 92.88% to 73.09%. Tail length(${\mu}m$) was increased as the dose of increased over 300 Gy. Growth characteristics (Germination rate, Survival rate, plant height, number of leaves, root length and fresh weight) were highly negatively ($P{\leq}0.01$) correlated with dose. While the tail length was highly positively ($P{\leq}0.01$) correlated with dose.

• Journal of Nutrition and Health
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• v.36 no.3
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• pp.255-261
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• 2003

Food irradiation has been steadily increasing in many countries in line with increasing international trade and concerns about naturally occurring harmful contaminants in food. Although irradiation provides an excellent safeguard for the consumer by destroying almost 100% of harmful bacteria, it is necessary to ensure the safety of irradiated foods. This study was performed to investigate the effect of an irradiated diet on lipid peroxidation in the plasma, liver, small intestinal mucosa, and lymphocyte DNA damage in mice. Eight-week old ICR mice were assigned to two groups to receive either non-irradiated or irradiated (10 kGy) diets containing 20.38% fish powder and 6.06% sesame seeds for 4 weeks. The resulting changes in the degrees of lipid peroxidation were evaluated based on the level of plasma and liver thiobarbituric acid reactive substance (TBARS), transmission electron micrograph of jejunal mucosa, and free radical-induced oxidative DNA damage in lymphocytes, as measured by alkaline comet assay (single cell gel electrophoresis). The peroxide values of the gamma irradiated diet were measured every week, and the sample for comet assay was taken at the end of the four week experimental period. There was no significant difference in food efficiency ratio between the two groups. The peroxide values of the diet were immediately increased to 35.5% after gamma irradiation and kept on increasing during storage. After 4 weeks, no differences in tissue or plasma TBARS value were observed between the two groups, but epithelial cells of jejumum showed osmiophillic laminated membranous structures, considered as myelin figures,. The oxidative DNA damage expressed as tail moment (TM) increased 30% in the blood lymphocytes of the mice fed the irradiated diet. In conclusion, the comet assay sensitively detected differences in lymphocyte DNA damage after feeding with the irradiated diet for 4 weeks. However, in order to ensure the safety of irradiated foods, it would be more useful to conduct a long-term feeding regimen using an irradiated diet and examine the level of lipid peroxidation and the state of oxidative stress in a greater range of organs.

• Parasites, Hosts and Diseases
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• v.51 no.2
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• pp.155-163
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• 2013

This study aimed to evaluate the efficacy of fructose-1,6-bis phosphate aldolase (SMALDO) DNA vaccination against Schistosoma mansoni infection using different routes of injection. The SMALDO has been cloned into the eukaryotic expression vector pcDNA3.1/V5-His TOPO-TA and was used in injecting Swiss albino mice intramuscularly (IM), subcutaneously (SC), or intraperitoneally (IP) ($50{\mu}g/mouse$). Mice vaccinated with non-recombinant pcDNA3.1 served as controls. Each group was immunized 4 times at weeks 0, 2, 4, and 6. Two weeks after the last booster dose, all mice groups were infected with 80 S. mansoni cercariae via tail immersion. At week 8 post-infection, animals were sacrificed for assessment of parasitological and histopathological parameters. High anti-SMALDO IgG antibody titers were detected in sera of all vaccinated groups (P<0.01) compared to the control group. Both the IP and SC vaccination routes resulted in a significant reduction in worm burden (46.2% and 28.9%, respectively, P<0.01). This was accompanied by a significant reduction in hepatic and intestinal egg counts (41.7% and 40.2%, respectively, P<0.01) in the IP group only. The number of dead eggs was significantly increased in both IP and IM groups (P<0.01). IP vaccination recorded the highest significant reduction in granuloma number and diameter (54.7% and 29.2%, respectively, P<0.01) and significant increase in dead miracidia (P<0.01). In conclusion, changing the injection route of SMALDO DNA vaccination significantly influenced the efficacy of vaccination. SMALDO DNA vaccination via IP route could be a promising protective and antipathology vaccine candidate against S. mansoni infection.

• Proceedings of the Korea Society of Environmental Toocicology Conference
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• 2003.10a
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• pp.159-159
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• 2003

Comet assay is a potential monitoring tool because DNA strand breakage may be produced by a wide range of agents. The comet assay, also called the single-cell gell electrophoresis (SCGE) assay, is rapid and sensitive method for the detection of DNA damage in cells. This study was performed for the identification of DNA damage in the cells from flounders and clams exposed to PAHs. As a control experiments, flounder and clam cells were exposed to $H_2O$$_2$. The cells exposed to $H_2O$$_2$ were displayed a typical nuclei movement DNA damage of cells were significantly increased when the isolated cells from the blood of flounders and the tissue of clams were in vitro exposed to the different concentrations (5, 10, 50, 100 ppb) of five kinds of PAHs (benzo[a]pyrene, pyrene, fluoranthene, anthrancene, and phenanthrene). For the in vivo test, flounders and clams were exposed to the different concentrations of BaP for 4 days. The results showed that DNA strand breakage was effected by the concentration of BaP and the duration of exposure. In high concentration of BaP, the mean tail lengths of nuclei was longer than it In low concentration, while the mean size of head DNA decreased. In this research, both in vitro and in vivo genotoxicity of PAHs could be biomonitored by the comet assay. Especially, clams and flounders seem to be useful as materials for monitoring genotoxic damage by comet assay.

• Animal Systematics, Evolution and Diversity
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• v.8 no.2
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• pp.231-242
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• 1992

Samples of two species of genus Ratts(black rat, Rattus rasttus Linnaeus; common rat, Rattus norvegicus Berkenhaut) in Korea were trapped and their 31 morphometric charcters were analyzed statistically in order to determine the range of geographic variation within each species and the interspecific differences. In addition, chromosomal G-bands and C-bands were compared and the fragment patterns of mtDNA resulted from the digestion with restriction enzymes were also analyzed. Samples of black rats from six localities in Korea were similar with one another in their morphometric characters: in head and body length, length of tail vertebrae, conventional karyotype and C-bands, they are comparable to Rattus rattus tanezumi in Japan. Specimens of common rats from seven localities in Korea were similar with one another in their morphometric characters: in conventional karyotype, they are comparable to Rattus norvegicus caraco in eastern Asia. Common rats differ from black rats in their morphometric characteris, chromosomal karyotypes and mtDNA. It is confirmed that correct species name of black rat in Korea is Rattus rattus tanezumi Tempminck: species name of common rat in Korea is Rattus norvegicus caraco Pallas: the common rat is a species, which is distinct from the black rat.

• Animal Systematics, Evolution and Diversity
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• v.32 no.4
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• pp.253-257
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• 2016

The genus Aphelenchoides (Fischer, 1894) includes a diverse group of species, some of which are of economic importance. A. bicaudatus (Imamura, 1931) Filipjev and Schuurmans Stekhoven, 1941 is reported for the first time from South Korea, with a detailed redescription of the species. Specimens were collected from chrysanthemum (Chrysanthemum morifolium) leaves and shoot tips in South Korea. The species was identified by morphological traits and molecular sequencing. A bifurcated tail distinguishes A. bicaudatus from its congeneric species. To confirm species identification, we determined the partial 18S ribosomal DNA sequence of the specimens and compared with those obtained from other Aphelenchoides species available on GenBank.