• Title/Summary/Keyword: DNA in tail

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Measurement of DNA Damage with Fpg/Endo III FLARE Assay and Real Time RT-PCR in SD Rats Exposed to Cumene

  • Kim, Soo-Jin;Rim, Kyung-Taek;Lee, Seong-Bae;Kim, Hyeon-Yeong
    • Molecular & Cellular Toxicology
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    • v.4 no.3
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    • pp.211-217
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    • 2008
  • To clarify the DNA damage from reactive oxygen species, we measured the DNA damage through Fpg/Endo III FLARE (Fragment Length Analysis with Repair Enzyme) assay and real time RT-PCR. The 80 SD rats assigned to 4 dose groups exposed to cumene vapor for 90 days. With Fpg/Endo III FLARE assay in hepatocytes, we found the OTM (Olive Tail Moment) and TL (Tail Length) significantly increased in no-enzyme treated and Fpg-treated control and 8 ppm groups with 28 days exposure. In Endo III-treated 8 ppm group, significantly increased the values with 90 days exposure. With lymphocytes, it was founded the values significantly increased in no-enzyme treated 800 ppm group in 28 and 90 days. It was significantly increased in Endo III-treated 80 ppm for 28 days and 800 ppm for 90 days. From the above findings, FLARE assay was suggested as being available as a biological marker for DNA damage induced by cumene exposure in SD rats. And we used real time RT-PCR for the OGG1 mRNA expression, it had dose-dependent biologic effects in 1 day exposure, but decrease the levels of rOGG1 mRNA. Our findings provide evidence that cumene exposure may cause suppression of rOGG1 in the rat hepatocytes or lymphocytes.

DNA Repair Capacity in Peripheral Blood Lymphocytes Predicts Efficacy of Platinum-based Chemotherapy in Patients with Gastric Cancer

  • Zhang, Yi-Yin;Gu, Kang-Sheng
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.9
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    • pp.5507-5512
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    • 2013
  • Objective: To investigate the correlation between ERCC1 expression levels in tumor tissue and peripheral blood lymphocytes (PBL) from patients with gastric cancer and assess the relationship between PBL DNA repair rate (DRR) and the efficacy of platinum chemotherapy. Methods: A total of 53 patients with gastric cancer receiving surgery and 20 controls were studied. ERCC1 protein expression in tumour tissue and PBL were determined by immunohistochemical staining. The PBL DRRs of 47 advanced patients and 20 controls were estimated by comet assay. Results: The positive expression rates of ERCC1 were 67. 9%, 56. 6% and 10.0% in tumour tissues, PBLs of gastric cancer patients, and PBLs of the control group. PBL ERCC1 expression correlated with that in tissue (${\chi}^2$=15. 463, p=0.000). Pearson contingency coefficient=0.475). DRRs of cancer patients by tail length (TL) (Z=4. 662, p=0.000) and tail moment (TM) (Z=3. 827, p=0.000) were significantly lower than that of control group. When TL was applied as an indicator, the correlation between DRR and chemotherapy efficacy was significant (Spearman rank correlation r=0.327, p=0.032). Patients with low levels of DRR in PBL presented better short-term efficacy of chemotherapy than those with high levels of DRR. Conclusions: The ERCC1 expression in PBLs may indirectly reflect ERCC1 expression in gastric cancer tissues. Compared with non-cancer populations, patients with gastric cancer may have lower DNA repair capacity. DRR in PBL may predict the short-term efficacy of platinum-based chemotherapy for patients with advanced gastric cancer.

Effect of Dendranthema indicum Extracts on Cell and DNA Damage Induced by Oxidative Stress (국화추출물이 산화적 스트레스에 의해 유발되는 세포와 DNA 손상에 미치는 영향)

  • Park, Young-Mi;Kim, Jee-In;Lee, Chang-Ho;Lim, Jae-Hwan;Seo, Eul-Won
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1698-1704
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    • 2011
  • In this study, we evaluated the protective effects of ethanol extracts from Dendranthema indicum on cell and DNA damages induced by oxidative stress. Antioxidant activities of D. indicum extracts are higher than scavenging activities of DPPH free radical and hydroxyl radical by 92.8% and 73.8%, respectively, and higher than ferrous iron chelating effects by 59.4%. D. indicum extracts showed a protective effect on oxidative cell damage by inhibiting lipid peroxidation by 90.3% in the control group, and inhibiting expression level of p21 protein by 79.6% for the control group. This means D. indicum extracts have a great protective effect against oxidative stress. DNA fragmentation inhibition in D. indicum extracts were 89.6% for the control group, which makes the movement of DNA tail reduced, and phosphorylation of H2AX was 20.2% of the radical experiment group. This means that D. indicum extracts effectively inhibit DNA fragmentation and H2AX phosphorylation. Taken together, we suggest that ethanol extract from D. indicum has a role as a useful chemopreventor against oxidative damage.

Antimutagenic Effects of Ginsenoside Rb$_1$, Rg$_1$ in the CHO-K1 Cells by Benzo[a]pyrene with Chromosomal Aberration Test and Comet Assay

  • Kim, Jong-Kyu;Kim, Soo-Jin;Rim, Kyung-Taek;Cho, Hae-Won;Kim, Hyeon-Yeong;Yang, Jeong-Sun
    • Molecular & Cellular Toxicology
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    • v.5 no.2
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    • pp.126-132
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    • 2009
  • The usage and types of chemicals are advancing, specializing, large-scaled increasing, and new chemical exposed workers are concerning to occupational disease. The generation of reactive oxygen in the body from carcinogen, mutation and DNA damage in cancer is protected by natural antioxidants (phytochemicals) with antimutagenic effect. There were many reports of ginsenoside Rb$_1$, Rg$_1$ grievances of the genetic mutation to suppress the effect confirm the genetic toxicity test with chromosomal aberration test and the Comet (SCGE) assay confirmed the suppression effect occurring chromosomal DNA damage. We had wanted to evaluate the compatibility and sensitivity between the chromosomal aberration (CA) test and the Comet assay. We used the CA test and Comet assay to evaluate the anti-genotoxicity of ginsenoside Rb$_1$ and Rg$_1$, in CHO-K1 (Chinese hamster ovary fibroblast) cell in vitro, composed negative control (solvent), positive control (benzo[a]pyrene), test group (carcinogen+variety concentration of ginsenoside) group. The positive control was benzo[a]pyrene (50 $\mu$M), well-known carcinogen, and the negative control was the 1 % DMSO solvent. The test group was a variety concentration of ginsenoside Rb$_1$, Rg$_1$ with 10$^{-8}$%, 10$^{-6}$%, 10$^{-4}$%, 10$^{-2}$%, 1%, 10%. In chromo-somal aberration test, we measured the number of cells with abnormally structured chromosome. In Comet assay, the Olive tail moment (OTM) and Tail length (TL) values were measured. The ratio of cell proliferation was increased 8.3% in 10$^{-8}$%, 10$^{-6}$%, 10$^{-4}$%, 10$^{-2}$%, 1%, 10% Rb$_1$ treated groups, and increased 10.4% in 10$^{-10}$%, 10$^{-8}$%, 10$^{-6}$%, 10$^{-4}$%, 10$^{-2}$%, 1% Rg$_1$ treated groups. In the CA test, the number of chromosomal aberration was decreased all the Rb$_1$ and Rg$_1$ treated groups. In the Comet assay, the OTM values were decreased in all the Rb$_1$ and Rg$_1$ treated groups. To evaluate the compatibility between CA and Comet assay, we compared the reducing ratio of chromosomal abnormalities with its OTM values, it was identified the antimutagenicity of ginsenoside, but it was more sensitive the CA test than the Comet assay. Ginsenoside Rb$_1$ and Rg$_1$ significantly decrease the number of cells with chromosomal aberration, and decrease the extent of DNA migration. Therefore, ginsenoside Rb$_1$, Rg$_1$ are thought as an antioxidant phytochemicals to protect mutagenicity. The in vitro Comet assay seems to be less sensitive than the in vitro chromosomal aberration test.

Mammary Gland Indices at the End of Lactation in the Superovulated Javanese Thin-Tail Ewes

  • Manalu, W.;Sumaryadi, M.Y.;Sudjatmogo, Sudjatmogo;Satyaningtijas, A.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.4
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    • pp.440-445
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    • 2000
  • Thirty lactating Javanese thin-tail ewes (12 ewes had been injected, prior to mating, with 700 IU pregnant mare serum gonadotropin, and 18 ewes with saline as a control) were used to evaluate the effect of superovulation on milk production during lactation and mammary chemical indices at the end of lactation. Thirteen ewes (9 control and 4 superovulated ewes) were fed at low and the other 17 ewes (9 control and 8 superovulated ewes) were fed at high quality ration. Superovulated ewes, either fed at low or high quality ration, had dramatically higher milk yields (57%). At the end of lactation, superovulated ewes had higher mammary dry fat-free tissue, mammary DNA concentration, total mammary DNA and RNA contents than nonsuperovulated ewes. Superovulation did not affect mammary RNA and collagen concentrations, and total collagen content. Ration quality did not significantly increase milk production during lactation and mammary chemical indices at the end of lactation. The observed increase in milk production in the superovulated ewes was probably due to the increased mammary secretory cell number and their synthetic activities during lactation as a result of the increased endogenous hormonal stimulation of mammary growth and development during pregnancy.

First Report of Two Plectus Species (Nematoda: Plectida) from Korea

  • Kim, Jiyeon;Yu, Jeong-Nam;Kim, Taeho
    • Animal Systematics, Evolution and Diversity
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    • v.36 no.3
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    • pp.251-257
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    • 2020
  • The genus Plectus Bastian, 1865 represents a group of free-living freshwater nematodes belonging to the family Plectidae Örley, 1880. However, only one species has been reported thus far from Korea. Plectus aquatilis Andrássy, 1985 and Plectus cirratus Bastian, 1865 are reported for the first time from Korea, from sediments collected from the Nakdong River. Plectus aquatilis is distinguished from other Plectus species by its three longitudinal alae in the lateral field, thin and directed cephalic setae, continuous lip region (head not set-off), and rectangular shaped tail. Plectus cirratus is distinguished from other Plectus species by its large body, two longitudinal alae in the lateral field, larger stoma, and longer tail. Morphological characters and measurements of the specimens generally agree with the original descriptions of Plectus species. Here, the morphological characters (detailed morphometrics) of two species in the genus Plectus are fully redescribed and illustrated using optical microscopy. DNA barcode sequence information from the 18S rDNA gene is also provided for molecular species identification and compared with sequences from other Plectus species available on GenBank.

Genetic Variability and Relationships of Native Japanese Chickens Assessed by Microsatellite DNA Profiling - Focusing on the Breeds Established in Kochi Prefecture, Japan -

  • Osman, S.A.-M.;Sekino, M.;Nishibori, M.;Yamamoto, Y.;Tsudzuki, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.6
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    • pp.755-761
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    • 2005
  • Blood samples were collected from eight native Japanese breeds of chickens (Miyadi-dori, Ohiki, Onaga-dori, Shoukoku, Tosa-Jidori, Tosa-Kukin, Toutenkou and Uzurao) and two foreign breeds of chickens (White Leghorn and Rhode Island Red) to examine the genetic variability and relationships among the breeds by using a microsatellite DNA technique. Except for the Shoukoku breed, the other Japanese chicken breeds all originate from Kochi Prefecture. Ohiki, Onaga-dori, Tosa-Jidori, Toutenkou and Uzurao are fancy fowl, and Miyadi-dori and Tosa-Kukin are utility fowl. Among the fancy fowl, Ohiki, Onaga-dori, and Toutenkou males have thick and long feathers in the saddle and tail. Genetic variabilities of the 20 microsatellites examined, varied depending on the breed: the mean number of alleles per locus ranged from 2.05 (Miyadi-dori) to 3.90 (Rhode Island Red); proportion of polymorphic loci ranged from 0.75 (Miyadi-dori) to 1.00 (Rhode Island Red, Shoukoku and Uzurao); and mean expected heterozygosity ranged from 0.330 (Miyadi-dori) to 0.607 (Rhode Island Red). Unique microsatellite alleles were detected in each breed. Using the neighbour-joining method, phylogenetic trees were constructed based on the genetic distances of D$_{A}$ and D$_{ST}$. Among the breeds originating from Kochi Prefecture, fancy and utility breeds belonged to different clusters. Among the fancy breeds, those having thick and long feathers in the tail and saddle showed a close genetic relationship to the Shoukoku breed, which also has thick and long feathers in the tail and saddle.

Molecular Biological Studies on Korean Garlic Viruses

  • Choi, Jin-Nam;Song, Jong-Tae;Shin, Chan-Seok;La, Yong-Joon;Lee, Jong-Seob;Choi, Yang-Do
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 1994.06a
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    • pp.86-102
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    • 1994
  • To understand the molecular structure and pathogenesis mechanism of Korean garlic viruses, we have isolate cDNA clones for garlic viruses. The partial nucleotide sequences of 24 cDNA clones were determined and that of six clones containing poly (A) tail were compared with those of other plant viruses. One of those clones, V9 has 81.8% similarity in nucleotide sequence and 93.0% in deduced amino acid sequence, respectively, to the coat protein gene for garlic mosaic virus (GMV). Northern blot analysis with the clone V9 demonstrated that the genome of GMV is 7.8 kb long and has poly (A) tail. The anti-coat protein antibody for GMV recognizes 35 kDa polypeptide which could be the coat protein of GMV from infected garlic leaf extract or virus preparation. Clone G7 has about 62% of deduced amino acid sequence identity with the members of potyvirus group. Northern blot analysis with the clone G7 demonstrated that the genome of the potyvirus I garlic is 9.0 kb long and has poly (A) tail. The third clone, S81, shows 42% amino acid identity to the potexvirus. The other clones are under the characterization. To test the possibility of producing garlic virus resistant plant, we have designed a hairpin type ribozyme to cleave V9 RNA at the middle of the coat protein gene. From the cleavage reactions in vitro with two different sizes of RNA substrates, V9SUB (144 nucleotides) and V9 RNA (1,361 nucleotides), the ribozyme can cleave V9 sequence effectively at the predicted site. To study the activity of the ribozyme in vivo, plant transformation is in progress. Further possibilities to produce garlic virus resistant plant will be discussed.

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Assessment of DNA Damage using an Alkaline Single Cell Gel Electrophoresis (SCGE) Comet Assay and Toxic Effects in Chickens by T-2 Toxin Treatment (T-2 toxin을 투여한 닭에서 Comet assay 방법을 이용한 DNA 손상 평가와 독성)

  • Hah Dae-Sik;Heo Jung-Ho;Lee Kuk-Cheon;Cho Myung-Heui;Kim Kuk-Hun;Kim Chung-Hui;Lue Jae-Du;Lee Seung-Hwan;Kim Gon-Sup;Kim Eui-Gyung;Kim Jong-Shu
    • Toxicological Research
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    • v.22 no.2
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    • pp.75-85
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    • 2006
  • This study was designed to evaluate the possible DNA damaging effects of T-2 toxin using an alkaline single cell gel electrophoresis (SCGE) comet assay and also to investigate toxic effects in chickens. A total of 20 chickens were used in these experiments. Graded concentrations of dietary T-2 toxin (0, 4, 8, and $16{\mu}g/g$ of diet) were given to groups of 5 broiler chickens. In comet assay, The DNA damage was analysed by the tail extent moment (TEM) and tail length (TL), which were used as markers of DNA strand breaks in SCGE. A significant dose-dependent increase in the extent of DNA migration as well as in the percentage of cells with tails was observed after treatment with T-2 toxin (P<0.05). Treatment with the low T-2 toxin ($4{\mu}/g$ of diet) induced a relatively low level of DNA damage in comparison with the high T-2 toxin ($16{\mu}/g$ of diet) group. The growth rate was significantly reduced by concentrations of 8, and $16{\mu}/g$ of diet (P < 0.05). The feed conversion ratio were significantly affected by any concentrations (P < 0.05). The relative weight of the spleen, and lung was decreased by the growth inhibitory concentrations. The bursa of Fabricius, thymus, and kid- ney were decreased in relative weight by concentrations of $16{\mu}/g$ of diet. The relative weight of the liver and heart were unaffected. The hemoglobin (Hb), hematocrit (HCT), and mean corpuscular hemoglobin (MCH) were decreased at concentration of $16{\mu}/g$ of diet. As compared with control chickens, there was no marked change in serum components except uric acid in T-2 treated chickens. All lymphoid tissues retained atrophic and lymphoid cell depletion throughout the three weeks trial.

Protective Effect of Kefir Grain Against Dextran Sodium Sulfate-Induced Colitis in Rats (Dextran Sodium Sulfate로 대장염을 유도한 흰쥐에서 캐피어 원말의 장보호 효과)

  • Ko, Young-Eun;Kim, Mi-Kyoung;Cho, Han-Young;Lee, In-Young;Ly, Sun-Yung
    • Journal of Nutrition and Health
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    • v.41 no.5
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    • pp.391-401
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    • 2008
  • Probiotics have emerged as a potential treatment modality for numerous gastrointestinal disorders, including IBD. However, few probiotics have undergone appropriate preclinical screening in vivo. Kefir is considered a probiotic, benefiting the host through its effects in the intestinal tract. Despite numerous studies examining the action of probiotics on the host organism, few have analyzed the effects on intestinal environment. We assessed the protective effect of kefir for three weeks before inducing colitis with 2% dextran sodium sulfate for five days. The DSS loads were similar in all DSS treatment group. The results of the experiment are as follows. Food intake and FER of experimental groups were not significantly different each other, but water consumption tended to be higher in all DSS treatment groups as compared with the normal control. And visual inspection of feces revealed mild diarrhea in rat given 2% DSS. The anti-inflammatory activity of kefir was determined by myeloperoxidase activity during the DSS treatment, and there was no significant difference in any group. The levels of thiobarbituric acid reactive substances (TBARS) as a colonic lipid peroxidation were significantly lower in the kefir intake groups than in rats treated with 2% DSS alone. The DNA % in tail and tail moment values as a DNA damage level of the blood lymphocytes in kefir intake groups tended to be lower than 2% DSS treatment alone, especially tail lengths were significantly diminished. According to the colonic histopathological assay, there were a severe inflammation of lamina propria and submucosa and mild edema in mucosa and sub mucosa in DSS alone treated group. We found a slight regenerative change in kefir treatment groups. In our experiments, this means that ulcerative colitis related to oxidative injury might be prevented by kefir as a probiotic. Further studies of the potential benefits of kefir as a probiotic in inflammatory condition are encouraged.