• Title/Summary/Keyword: Damaged DNA

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Observation of Mitotic Chromosome behavior according to Different Treatment Methods of DNA Methylation Inhibitor

  • Seong-Wook Kang;Ji-Yoon Han;Seong-Woo Cho
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.221-221
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    • 2022
  • Chromosome breakage occurred by DNA methylation inhibitor. Zebularine is known as DNA methylation inhibitor and suitable for water solubility among different DNA methylation inhibitors as 5-Azacytidine and 5-aza-2'-deoxycytidine. We used zebularine as mutagen according to different methods by roots absorption and seed imbibition. After zebularine treatment, DNA methylation inhibitor, we observed mitotic chromosome behavior what is different according to two different treatment methods. First, seed imbibition treatment in 1,000 μM of zebularine solution for 72 hours in dark conditions. The second treatment to seedlings of Keumkang was also treated in 1,000 μM of zebularine solution for 72 hours after germination. Root and shoot showed different elongations in each treatment. Root absorption treatment(3.01±0.48, 2.00±0.26) showed the shortest elongation in root and shoot than control(8.16±0.61, 4.03±0.48) and seed imbibition treatment(4.33±0.80, 2.48±0.36). It can be explained root tip meristematic cell activity was damaged by DNA methylation inhibitor. Primary root tips were collected in DW for 24 hours at low temperature(0℃) and fixed in fixation solution for 3 days to chromosome observation in mitosis. Mitotic index, chromosome structure and chromosome aberration were observed by phase-contrast microscope. Mitotic index of the control(0.29) showed twice mitotic cells as the treated groups(imbibition 0.15, absorption 0.14). Observation of chromosomes showed some short chromosomes and loosen chromosomes affected by zebularine. It is considered because of zebularine damage DNA in mitosis. We observed "gap by chromosome breakage" in chromosomes that have loose parts between centromere and telomere. It seems demethylation of zebularine occurs chromosome breakage.

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Effect of forensic short wavelength UV on DNA (법과학 단파자외선이 DNA에 미치는 영향)

  • Kim, A-Ram;Kim, Woo-Joong;Park, Hee-Yeon;Lim, Si-Keun
    • Analytical Science and Technology
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    • v.27 no.3
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    • pp.147-152
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    • 2014
  • RUVIS(Reflective Ultraviolet Imaging System) is an effective equipment that detects the location of latent fingerprint at crime scene using short wavelength ultraviolet of 254 nm. In this study, the degree of DNA damage in biological samples was compared depending on the distance and time of processing using four commonly used RUVIS. 50% of DNA was damaged by treating 10 seconds at 10 cm distance in 3 types of RUVIS such as Police RUVIS, SIRCHIE mini light and SIRCHIE RUVIS. In addition, the degree of DNA damage was increased as the distance was closer and the treatment time was longer. It showed that short wavelength UV could cause DNA damage when used close to the samples at crime scene. Therefore, it was suggested to use RUVIS at a distance of at least 1 m. The degree of DNA damage was not significant by Polilight which used long wavelength ultraviolet of 350 nm. As a result, the choice and usage of which UV light and RUVIS were critical for detection of fingerprint and successful DNA typing.

The Role of Active Oxygen on DNA Damage by Linoleic Acid Peroxidation Products (Linoleic acid 산화생성물(酸化生成物)의 DNA손상작용에 있어서의 활성산소종(活性酸素種)의 역할)

  • Kim, Seon-Bong;Kang, Jin-Hoon;Lee, Yong-Woo;Kim, In-Soo;Park, Yeung-Ho
    • Korean Journal of Food Science and Technology
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    • v.19 no.4
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    • pp.311-316
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    • 1987
  • The present paper was carried out to investigate the effects of active oxygen radicals on the DNA damage by linoleic acid peroxidation by using active oxygen scavengers in a linoleic acid-DNA system. DNA was greatly damaged by linoleic acid peroxidation, and the DNA damage was inhibited by the addition of active oxygen scavengers. Among active oxygen scavengers tested, ${\alpha}-tocopherol$ and superoxide dismutase greatly inhibited the DNA damage, but catalase and tris (hydroxymethyl) aminomethane didn't show such effects. Accordingly, singlet oxygen and superoxide anion greatly affected to the DNA damage occurring during linoleic acid peroxidation, and hydrogen peroxide was shown to participate in DNA damage in the early stage of peroxidation. And, the DNA damage by active oxygen radicals was mainly induced in the early stage of linoleic acid peroxidation.

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Mal-differentiation of Stem Cells: Cancer and Ageing (줄기세포의 분화 결손으로 인한 노화와 암화)

  • Lee, Mi-Ok;Cha, Hyuk-Jin
    • KSBB Journal
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    • v.26 no.3
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    • pp.183-188
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    • 2011
  • Adult stem cells, which have characteristic of self-renewal and multipotency, are specialized cell types, responsible for the tissue regeneration of the damaged tissue. Recent studies suggest that stem cells senescence (or stem cells' ageing) is closely associated with the variety of ageing-related phenotypes such as tissue atrophy, degenerative diseases and onset of cancers. During ageing, declining of stem cells function and subsequently occurring mal-differentiation of stem cells would be important to understand the biological process of development of ageing-related phenotypes such as tissue degenerations and cancers. This review focuses on the DNA damage stress as a cause of senescence of stem cells and their mal differentiation, which is closely link to defect of regeneration potentials and neoplastic transformation. Understanding of molecular mechanisms governingsuch events is likely to have important implications for developing novel avenues for balancing tissue homeostasis longer period of time, further leading to 'Healthy ageing'.

Comparison of Endonuclease-Sensitive Sites by T4 Endonuclease V and UvrABC Nuclease Treatments Followed by Formamide or Sodium Hydroxide Denaturation

  • Chang, Yung-Jin
    • BMB Reports
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    • v.31 no.4
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    • pp.405-408
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    • 1998
  • Endonuclease-sensitive sites detected by T4 endonuclease V or UvrABC nuclease treatments were compared in the dihydrofolate reductase gene of UV-irradiated Chinese hamster ovary B-11 cells. The number of endonuclease-sensitive sites detected by T4 endonuclease V treatment followed by NaOH denaturation was twice that of formamide denaturation. Repeated treatment of damaged genomic DNA with T4 endonuclease V resulted in no further increase in the number of endonuclease-sensitive sites detected. The numbers of endonuclease-sensitive sites detected by UvrABC nuclease using each denaturation condition were similar. Sequential treatment with the two endonucleases using formamide denaturation resulted in twice the number of endonuclease-sensitive sites detected by treatment of each nuclease alone. Due to a lack of AP endonuclease activity these results suggest the presence of T4 endonuclease V-sensitive sites which could be complemented by alkaline gel separation or by UvrABC nuclease treatment.

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Backbone Assignment of the N-terminal Domain of Human Replication Protein A 70 kDa

  • Lee, Sungjin;Park, Chin-Ju
    • Journal of the Korean Magnetic Resonance Society
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    • v.20 no.4
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    • pp.138-142
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    • 2016
  • Replication Protein A (RPA) is the eukaryotic single-stranded DNA binding protein. It involves in DNA replication, repair, and damage response. Among three subunits, RPA70 has a protein-protein binding domain (RPA70N) at the N-terminal. It has known that the domain recruits several damage response proteins to the damaged site. Also, it is suggested that there are more candidates that interact with RPA70N. Even though several studies performed on the structural aspects of RPA70N and its ligand binding, the backbone assignments of RPA70N is not available in public. In this study, we present the backbone assignments of RPA70N.

Apoptosis by Direct Current Treatment in Tumor Cells and Tumor Tissues

  • Kim, H. B.;S. B. Sim;S. Ahn
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.48-48
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    • 2003
  • Electric field induces cell fusion, electroporation on biological cells, including apoptosis. Apoptosis is expressed in a series of natural enzymatic reactions for the natural elimination of unhealthy, genetically damaged, or otherwise aberrant cels that are not needed or not advantageous to the well-being of the organism. Its markers involve cell shringkage, activation of intracellular caspase proteases, externalization of phosphatidylserine at the plasma membrane, and fragmentation of DNA.

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Intra- and Extra-cellular Mechanisms of Saccharomyces cerevisiae Inactivation by High Voltage Pulsed Electric Fields Treatment (고전압 펄스 전기장에 의한 Saccharomyces cerevisiae의 세포내·외적 사멸 기작 연구)

  • Lee, Sang-Jae;Shin, Jung-Kue
    • Korean Journal of Food Science and Technology
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    • v.47 no.1
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    • pp.87-94
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    • 2015
  • High voltage pulsed electric fields (PEF) treatment is one of the more promising nonthermal technologies to fully or partially replace thermal processing. The objective of this research was to investigate the microbial inactivation mechanisms of PEF treatment in terms of intra- and extracellular changes in the cells. Saccharomyces cerevisae cells treated with PEF showed cellular membrane damage. This resulted in the leakage of UV-absorbing materials and intracelluar ions, which increased with increasing treatment time and electric fields strength. This indicates that PEF treatment causes cell death via membrane damage and physical rupture of cell walls. We further confirmed this by Phloxine B staining, a dye that accumulates in dead cells. Using scanning and transmission electron microscopy, we observed morphological changes as well as disrupted cytoplasmic membranes in PEF treated S. cerevisae cells. In addition, PEF treatment led to damaged chromosomal DNA in S. cerevisiae.

Coiled-Coil Domain-Containing Protein 98 (CCDC98) Regulates Cyclin B1 Expression by Affecting WTAP Protein Stability (WTAP 단백질의 안정성을 통한 CCDC98 단백질의 cyclin B1 발현 조절)

  • Oh, Yun-Jung;Lee, Eun-Hee;Lee, Il-Kyu;Kim, Kyung-Soo;Kim, Hong-Tae
    • Journal of Life Science
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    • v.21 no.8
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    • pp.1067-1075
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    • 2011
  • Coiled-coil domain-containing protein 98 (CCDC98) plays a role in G2/M DNA damage checkpoint pathways by recruiting breast cancer 1 (BRCA1)-A complex to the DNA-damaged sites. However, the molecular mechanism of CCDC98 on the DNA damage-induced G2/M checkpoint pathways is unclear. In this study, we identifed Wilms tumor 1-associating protein (WTAP) as a novel CCDC98-binding protein, using tandem affinity purification. We confirmed the association between CCDC98 and WTAP using in vivo and in vitro binding assays. We demonstrated that CCDC98 regulates cyclin B1 expression by affecting WTAP protein stability. Based on these results, we suggest that CCDC98 may act as a novel cell cycle regulator by regulating the expression level of cyclin B1.

Efficiency of PDNR (Polydeoxyribonucleotide) extraction from various plant species and its in vitro wound healing activity (다양한 식물에서의 PDRN(Polydeoxyribonucleotide) 추출 수율 비교 및 상처치유 효능 분석)

  • Song, Mi-Hee;Choi, Moon-Hyeok;Jeong, Jin-Hyoung;Lee, Sang-Sik;Jeong, Woo-Young
    • The Journal of Korea Institute of Information, Electronics, and Communication Technology
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    • v.15 no.5
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    • pp.387-395
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    • 2022
  • PDRN (Polydeoxyribonucleotide) is a DNA-derived polymer that promotes self-renewal of damaged cells and tissues as a tissue regeneration active material. PDRN is a DNA fragment cut into small sizes by various physical or chemical methods. When administered to the body, PDRN binds and stimulates the adenosine A2A receptor on the surface of tissue cells to promote cell regeneration, accelerate wound healing, and reduce pain. Although PDRN is prepared from testis or semen of fish in most cass, PDRN extraction from various plants species was performed in the present study. Among 7 tested plant species, the highest DNA yield and purity was obtained form mugwort (Chrysanthemum coronarium, C.c), followed by broccoli (Brassica oleracea, B.o). Then, we evaluated the in vitro wound healing capacity of PDRNs prepared from these two selected plants. PDRN from C.c and B.o. significantly stimulated the wound healing process at ㎍/ml range. The present study suggests that PDRN from plant species can be an effective alternative to PDRN from marine organism.