• Title/Summary/Keyword: mDNA

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The Genetic Analysis Study of Ancient Human Bones Excavated at Janggi-dong site, Gimpo (김포 장기동 유적 출토 인골의 유전자 분석 연구)

  • Seo, Min Seok;Cho, Eun Min;Kim, Yun Ji;Kim, Sue Hoon;Kang, So Yeong
    • Journal of Conservation Science
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    • v.30 no.4
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    • pp.409-416
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    • 2014
  • Most human bones of Joseon Dynasty period are so good condition that we can do research in physical anthropology, genetics and chemistry with them. In this study, we analyzed DNA typing using 6 human bones of Joseon dynasty period excavated at Janggi-dong, Gimpo. The DNA typing was mitochondrial DNA haplotype, Y-chromosome haplotype and sex determination. Prior to DNA analysis, we distinguished histological index of 6 human bones. As the result of mitochondrial DNA analysis, most of bones were confirmed as haplogroup G, R11, M7, A5, etc. As the result of sex determination, 4 human bones were female and 2 human bones were male. The male haplogroup was confirmed as haplogroup O by the single nucleotide polymorphism analysis of Y chromosome. For extensive ancient human bone analysis, researchers need to apply a histological index to select ancient human bones and explain a relationship among ancient human bones with various analyses of mitochondrial and nuclear DNA.

Lymphocyte DNA Damage and Anti-Oxidative Parameters are Affected by the Glutathione S-Transferase (GST) M1 and T1 Polymorphism and Smoking Status in Korean Young Adults (흡연 여부에 따른 Glutathione S-transferase (GST) M1 및 T1 유전자 다형성이 우리나라 젊은 성인의 임파구 DNA 손상과 항산화 영양상태 지표들 간의 관련성에 미치는 영향)

  • Han, Jeong-Hwa;Lee, Hye-Jin;Kang, Myung-Hee
    • Journal of Nutrition and Health
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    • v.44 no.5
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    • pp.366-377
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    • 2011
  • Glutathione S-transferase (GST) is a multigene family of phase II detoxifying enzymes that metabolize a wide range of exogenous and endogenous electrophilic compounds. GSTM1 and GSTT1 gene polymorphisms may account for inter-individual variability in coping with oxidative stress. We investigated the relationships between the level of lymphocyte DNA and antioxidative parameters and the effect on GST genotypes. GSTM1 and GSTT1 were characterized in 301 young healthy Korean adults and compared with oxidative stress parameters such as the level of lymphocyte DNA, plasma antioxidant vitamins, and erythrocyte antioxidant enzymes in smokers and non smokers. GST genotype, degree of DNA damage in lymphocytes, erythrocyte activities of superoxide dismutase, catalase, and glutathione peroxidase (GSH-Px), and plasma concentrations of total radical-trapping antioxidant potential (TRAP), vitamin C, ${\alpha}$- and ${\gamma}$-tocopherol, ${\alpha}$- and ${\beta}$-carotene, and cryptoxanthin were analyzed. Lymphocyte DNA damage assessed by the comet assay was higher in smokers than that in non-smokers, but the levels of plasma vitamin C, ${\beta}$-carotene, TRAP, erythrocyte catalase, and GSH-Px were lower than those of non-smokers (p < 0.05). Lymphocyte DNA damage was higher in subjects with the GSTM1- or GSTT1-present genotype than those with the GSTM1-present or GSTT1- genotype. No difference in erythrocyte antioxidant enzyme activities, plasma TRAP, or vitamin levels was observed in subjects with the GSTM1 or GSTT1 genotypes, except ${\beta}$-carotene. Significant negative correlations were observed between lymphocyte DNA damage and plasma levels of TRAP and erythrocyte activities of catalase and GSH-Px after adjusting for smoking pack-years. Negative correlations were observed between plasma vitamin C and lymphocyte DNA damage only in individuals with the GSTM1-present or GSTT1- genotype. The interesting finding was the significant positive correlations between lymphocyte DNA damage and plasma levels of ${\alpha}$-carotene, ${\beta}$-carotene, and cryptoxanthin. In conclusion, the GSTM1- and GSTT1-present genotypes as well as smoking aggravated antioxidant status through lymphocyte DNA damage. This finding confirms that GST polymorphisms could be important determinants of antioxidant status in young smoking and non-smoking adults. Consequently, the protective effect of supplemental antioxidants on DNA damage in individuals carrying the GSTM1- or GSTT1-present genotypes might show significantly higher values than expected.

Alterations in Mitochondrial DNA Copy Numbers and Mitochondrial Oxidative Phosphorylation (OXPHOS) Protein Levels in Gastric Cancer Tissues and Cell Lines (위암 조직과 세포주에서 mDNA와 OXPHOS 단백질 분석)

  • Siregar, Adrian;Hah, Young-Sool;Moon, Dong Kyu;Woo, Dong Kyun
    • Journal of Life Science
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    • v.31 no.12
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    • pp.1057-1065
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    • 2021
  • Alterations in mitochondrial DNA (mtDNA) copy numbers have been reported in patients with stomach cancer and suggested to play a role in gastric carcinogenesis or gastric cancer progression. However, changes in the levels of mitochondrial proteins or mtDNA-encoded oxidative phosphorylation (OXPHOS) proteins in gastric cancer remain unclear. In this study, we investigated mtDNA contents, mitochondrial protein levels, and mtDNA-encoded OXPHOS protein levels in gastric cancer tissues and cell lines. We correlated mtDNA copy numbers with clinicopathologic features of the gastric cancer samples used in this study and used quantitative PCR to analyze the mtDNA copy numbers of the gastric cancer tissues and cell lines. Western blot analysis was used for assessing the amounts of mitochondrial proteins and mtDNA-encoded OXPHOS proteins. Among the 27 gastric cancer samples, 22 showed a reduction in mtDNA copy numbers. The mtDNA content was increased in the other five samples relative to that in normal matched gastric tissues. Mitochondrial protein and OXPHOS protein levels were reduced in some gastric cancer tissues. However, mitochondrial protein and OXPHOS protein levels in gastric cancer cell lines were not always in line with their mtDNA contents. The mtDNA copy numbers were reduced in five gastric cancer cell lines tested in this study. In summary, this study reports a common reduction in mtDNA contents in gastric carcinoma tissues and cell lines, pointing to the possible involvement of mtDNA content alterations in tumorigenesis of the stomach.

Characterization of Albino Tobaccos (Nicotiana tabacum L.) Derived from Leaf Blade-Segments Cultured in vitro

  • Bae, Chang-Hyu;Tomoko Abe;Lee, Hyo-Yeon;Kim, Dong-Cheol;Min, Kyung-Soo;Park, Kwan-Sam;Tomoki Matsuyama;Takeshi Nakano;Shigeo Yoshida
    • Journal of Plant Biotechnology
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    • v.1 no.2
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    • pp.101-107
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    • 1999
  • The leaf blade-segments of albino tobacco (Nicotiana tabacum L.) were cultured on MS media containing different concentrations of BAP (0, 0.4, 2.2, 4.4, 22.2 ${\mu}{\textrm}{m}$) with or without NAA (0, 0.5, 2.7 ${\mu}{\textrm}{m}$). Multiple shoots were induced on the media containing 0.4 to 2.2 ${\mu}{\textrm}{m}$ BAP. The best condition for multiple shoot induction with root formation was MS media containing 4.4 ${\mu}{\textrm}{m}$ BAP and 0.5 ${\mu}{\textrm}{m}$ NAA. The regenerated albino plants showed a significant reduction in accumulation of chlorophylls and carotenoids. The drastic reduction of the pigments content was associated with the distinct alterations in gene expression in the albino plants. firstly, the expression of plastid genes, such as rbcL, psbA, 165 rDNA and 235 rDNA, was reduced at the level of transcripts in the regenerated albino plants. Secondly, the alteration of structure of the plastid genes was not detected in the albino plants. However, the copy number of the plastid genes whose transcription level was reduced greatly was increased approximately two-fold, although the transcriptions of nuclear gene (255 rDNA) showed the wild-type level.

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The Repair of MNNG-Induced DNA Damage and Its Relation to Chromosome Aberrations in Mammalian Cells (MNNG에 의한 DNA 회복합성과 염색체 이상과의 연관성에 관한 연구)

  • Kim, Choon-Kwang;Lee, Chun-Bok
    • The Korean Journal of Zoology
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    • v.23 no.3
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    • pp.115-123
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    • 1980
  • The rates of escision repair at various doses and times after MNNG treatment in CHO cells were compared with the frequencies of chromosome aberrations to determine a possible relation between there two types of biological phenomena, and the results obtained were as follows: 1. the MNNG-induced excision repair was dose-dependent in te ranges between $0.5 \\times 10^-5$M. The maximum rate of excision repair was occurred in the cells soon after the treatment. The rates were then gradually decreased and appeared about 66% of 0 hour at 24 hours. 2. The rates of chromosome aberrations induced by MNNG was the highest at 6 hours, in which majority were chromatid deletions. The rates of chromatid deletions decreased, whereas chromatid exchanges increased with time, resulting is about equal rates at 24 hours after treatment. 3. The rates of excision repair at different times after MNNG treatment were roughly related to the total breaks per cell. The rates, however, did not show any relation to either chromatid exchanges or deletions. These results may suggest that excision repair may not be directly related to chromosome aberrations in MNNG treated CHO cells.

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Toxicological Effects of B(a)P on Preimplantation Mouse Embryos in Vitro (in vitro에서 B(a)P이 착상전 마우스 배자에 미치는 독성학적 영향에 관한 연구)

  • 박귀례;이유미;김판기;신재호;강태석;김주일;장성재
    • Journal of Environmental Health Sciences
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    • v.24 no.2
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    • pp.126-133
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    • 1998
  • Effects of B(a)P on preimplantation mouse embryos in vitro were studied. Preimplantation mouse embryos were exposed to a concentration of 0.3, 1, 3 and 10 $\mu$M B(a)P for 72 hrs. The toxicological effects of B(a)P were evaluated by morphological observation of embryos up to the blastocyst stage, and by measuring DNA, RNA and protein synthesis by radioactive precursor incorporation. At 1 $\mu$M B(a)P did not affect preimplantation development but interfered with hatching and ICM formation. Suppressing effect of ICM formation was dose dependent. At the eight cell stage, the developmental rate was decreased at above 3 $\mu$M of B(a)P. At the blastocyst stage, attachment and trophoblast outgrowth were diminished at the 10 $\mu$M of B(a)P and ICM formation was decreased at 1 $\mu$M of B(a)P. Inner cell number of blastocyst was decreased dose dependently. So, number of ICM was one of the most sensitive and toxicological end point. The RNA incorporation rate of 0.1 $\mu ^3$H-uridine was dosedependent and the protein incroporation of 0.5 $\mu Ci ^{35}$S-methionine showed a significant decrease after 48 hrs. But the DNA incorporation rate of methyl-$^3$H thymidine was not affected. Our results suggested that B(a)P did not affect the DNA replication but transcription was inhibited by dose dependent manner. There delay of development during the blastocyst stage was mainly due to the inhibition of RNA synthesis followed by protein synthesis.

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Generation of Reactive Oxygen Species and Subsequent DNA Fragmentation in Bovine Cultured Somatic Cells

  • Hwang, In-Sun;Kim, Ho-Jeong;Park, Chun-Keun;Yang, Boo-Keun;Cheong, Hee-Tae
    • Reproductive and Developmental Biology
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    • v.35 no.4
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    • pp.485-489
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    • 2011
  • The present study was conducted to examine the reactive oxygen species (ROS) generation levels and subsequent DNA damage in the bovine cultured somatic cells. Bovine ear skin cells were classified by serum starvation, confluence and cycling cells. Cells were stained in 10 ${\mu}M$ dichlorohydrofluorescein diacetate ($H_2DCFDA$) or 10 ${\mu}M$ hydroxyphenyl fluorescein (HPF) dye to measure the $H_2O_2$ or $^{\cdot}OH$ radical levels. The samples were examined with a fluorescent microscope, and fluorescence intensity was analyzed in each cell. $H_2O_2$ and $^{\cdot}OH$ radical levels of cultured somatic cells were high in confluence group ($7.1{\pm}0.7$ and $8.4{\pm}0.4$ pixels/cell, respectively) and significantly low in serum starvation group ($4.9{\pm}0.4$ and $7.0{\pm}0.4$ pixels/cell, respectively, p<0.05). Comet tail lengths of serum starvation ($148.3{\pm}5.7$ ${\mu}M$) and confluence ($151.1{\pm}5.0$ ${\mu}M$) groups were found to be significantly (p<0.05) increased in comparison to that of cycling group ($137.1{\pm}7.5$ ${\mu}M$). These results suggest that the culture type of donor cells can affect the ROS generation, which leads the DNA fragmentation of the cells.

The Inhibitory Effects of Ahnjeonbaekho-tang on FRTL-5 Cell Proliferation and Thyroxine Synthesis

  • Kang, Shin-Ik;Lee, Byung-Cheol;Ahn, Young-Min;Doo, Ho-Kyung;Ahn, Se-Young
    • The Journal of Internal Korean Medicine
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    • v.27 no.3
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    • pp.653-663
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    • 2006
  • Objective : Graves' disease, the most common cause of hyperthyroidism, is an autoimmune disorder associated with autoantibodies to the TSH receptor. The clinical features of Graves' disease are goiter and hypermetabolic symptoms induced by excessive hormones. Antithyroid drug therapy is the first-line treatment for Graves' disease in Korea, Japan and European countries. Yet in spite of a long period and high-dose of treatment, it is hard to achieve remission because of adverse effects, frequent recurrence and resistance to antithyroid drugs. Recently, it has been reported that the abnormal thyroid hormone and clinical symptoms of Graves' disease were reduced by Ahnjeonbaekho-tang (AJBHT). Methods : To investigate the effectiveness and action mechanism of AJBHT, we studied the influence of AJBHT on FRTL-5 thyroid cell proliferation, DNA synthesis and expression of T4, TSH, cAMP, Tg and TPO mRNA. Results : AJBHT significantly inhibited the FRTL-5 cell proliferation, DNA synthesis, T4 synthesis, cAMP production and the expression of Tg mRNA in comparison with control and MMI. Conclusions : These results suggest that AJBHT may inhibit the cell proliferation and DNA synthesis by regulating the cAMP, and suppress the T4 synthesis by modulating Tg mRNA expression and cAMP synthesis, and that it may be useful agent for treating the goiter and hormone abnormality of Graves' disease.

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Sequencing of cDNA Clones Expressed in Adipose Tissues of Korean Cattle

  • Bong, J.J.;Tong, K.;Cho, K.K.;Baik, M.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.4
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    • pp.483-489
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    • 2005
  • To understand the molecular mechanisms that regulate intramuscular fat deposition and its release, cDNA clones expressed in adipose tissues of Korean cattle were identified by differential screening from adipose tissue cDNA library. By partial nucleotide sequencing of 486 clones and a search for sequence similarity in NCBI nucleotide databases, 245 clones revealed unique clones. By a functional grouping of the clones, 14% of the clones were categorized to metabolism and enzyme-related group (stearoyl CoA desaturase, lactate dehydrogenase, fatty acid synthase, ATP citrate lyase, lipoprotein lipase, acetyl CoA synthetase, etc), and 6% to signal transduction/cell cycle-related group (C/EBP, cAMP-regulated phosphoprotein, calmodulin, cyclin G1, cyclin H, etc), and 4% to cytoskeleton and extracellular matrix components (vimentin, ankyrin 2, gelosin, syntenin, talin, prefoldin 5). The obtained 245 clones will be useful to study lipid metabolism and signal transduction pathway in adipose tissues and to study obesity in human. Some clones were subjected to full-sequencing containing open reading frame. The cDNA clone of bovine homolog of human prefoldin 5 gene had a total length of 959 nucleotides coding for 139 amino acids. Comparison of the deduced amino acid sequences of bovine prefoldin 5 with those of human and mouse showed over 95% identity. The cDNA clone of bovine homolog of human ubiquitin-like/S30 ribosomal fusion protein gene had a total length of 484 nucleotides coding for 133 amino acids. Comparison of the deduced amino acid sequences of bovine ubiquitin-like/S30 ribosomal fusion protein gene with those of human, rat and mouse showed over 97% identity. The cDNA clone of bovine homolog of human proteolipid protein 2 mRNA had a total length of 928 nucleotides coding for 152 amino acids. Comparison of the deduced amino acid sequences of bovine proteolipid protein 2 with those of human and mouse showed 87.5% similarity. The cDNA clone of bovine homolog of rat thymosin beta 4 had a total length of 602 nucleotides coding for 44 amino acids. Comparison of the deduced amino acid sequences of bovine thymosin beta 4 gene with those of human, mouse and rat showed 93.1% similarity. The cDNA clone of bovine homolog of human myotrophin mRNA had a total length of 790 nucleotides coding for 118 amino acids. Comparison of the deduced amino acid sequences of bovine myotrophin gene with those of human, mouse and rat showed 83.9% similarity. The functional role of these clones in adipose tissues needs to be established.

Study on DNA Content and Ki-67 Antibody Expression by Means of Image Analyzer for the Benign and Malignant Lesions of the Larynx (후두 편평상피의 전암성 및 악성병변에서 화상분석기를 이용한 DNA 배수성검사와 Ki-67 항체 양성세포의 분석에 관한 연구)

  • 주형로;이선희;최종욱;김인선
    • Proceedings of the KOR-BRONCHOESO Conference
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    • 1993.05a
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    • pp.89-89
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    • 1993
  • The laryngeal epithelial cell kinetics of 26 laryngeal lesions(invasive squamous cell carcinoma 14, epithelial hyperplasia 5, laryngeal nodule 7) were studied by immunehistochemical analysis with the monoclonal antibody Ki-67, which reacts with nuclear antigen in proliferating cells using paraffin embedded tissue. For DNA analysis, touch implint with fresh biopsy specimens were stained with feulgen and analyzed by image analyzer in 22 cases. 1) The proportion of Ki-67-positive cells were 32.65$\pm$ 11.59% in invasive squamous cell ca, 20.14$\pm$3.38% in epithelial hyperplasia lesion and 11.66$\pm$3.02% in laryngeal nodule. 2) DNA aneuploidy was found in 7 cases of 10(70%) invasive squamous cell carcinomas, 2 cases of 5(40%) epithelial hyperplasia lesions and all cases of laryngeal 3) Proliferation index(S phase+G2/M phase) show 23.42$\pm$11.33% in squamous cell carcinoma, 13.09$\pm$ 10.90% in epithelial hyperplasia lesion and 4.50$\pm$1.19% in laryngeal nodule. As the results, measuring the DNA content from touch imprint method together positivity of Ki-67 antibody from the microtissue during the laryngeal microscopic surgery, cell kinetics can be assessed as an effort of deciding the prognosis and provide a key to the management of precancerous lesions.

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