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Inhibition of DNA-dependent Protein Kinase by Blocking Interaction between Ku Complex and Catalytic Subunit of DNA-dependent Protein Kinase

  • Kim, Chung-Hui;Cuong, Dang-Van;Kim, Jong-Su;Kim, Na-Ri;Kim, Eui-Yong;Han, Jin
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.1
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    • pp.9-14
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    • 2003
  • Recent studies indicated that cancer cells become resistant to ionizing radiation (IR) and chemotherapy drugs by enhanced DNA repair of the lesions. Therefore, it is expected to increase the killing of cancer cells and reduce drug resistance by inhibiting DNA repair pathways that tumor cells rely on to escape chemotherapy. There are a number of key human DNA repair pathways which depend on multimeric polypeptide activities. For example, Ku heterodimer regulatory DNA binding subunits (Ku70/Ku80) on binding to double strand DNA breaks (DSBs) are able to interact with 470-kDa DNA-dependent protein kinase catalytic subunit (DNA-PKcs), and are essential for DNA-dependent protein kinase (DNA-PK) activity. It has been known that DNA-PK is an important factor for DNA repair and also is a sensor-transmitting damage signal to downstream targets, leading to cell cycles arrest. Our ultimate goal is to develop a treatment of breast tumors by targeting proteins involved in damage-signaling pathway and/or DNA repair. This would greatly facilitate tumor cell cytotoxic activity and programmed cell death through DNA damaging drug treatment. Therefore, we designed a domain of Ku80 mutants that binds to Ku70 but not DNA end binding activity and used the peptide in co-therapy strategy to see whether the targeted inhibition of DNA-PK activity sensitized breast cancer cells to irradiation or chemotherapy drug. We observed that the synthesized peptide (HNI-38) prevented DNA-PKcs from binding to Ku70/Ku80, thus resulting in inactivation of DNA-PK activity. Consequently, the peptide treated cells exhibited poor to no DNA repair, and became highly sensitive to IR or chemotherapy drugs, and the growth of breast cancer cells was inhibited. Additionally, the results obtained in the present study also support the physiological role of resistance of cancer cells to IR or chemotherapy.

Adaptive Response Induced by Low Dose Ionizing Raditation in Human Cervical Carcinoma Cells

  • Kim, Jeong -Hee;Lee, Kyung -Jong;Cho, Chul -Koo;Yoo, Seong -Yul;Kim, Tae -Hwan;Ji, Young -Hoon;Kim, Sung -Ho
    • Archives of Pharmacal Research
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    • v.18 no.6
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    • pp.410-414
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    • 1995
  • Adaptive response induced by low dese .gamma.-ray irradiation in human cervical carcinoma cells was examined. Cells were exposured to low dose of .gamma.-ray irradiation in human cervical carcinoma cells was examined. Cells were exposured to low dose of .gamma.-ray (1-cGy) followed by high doses of r-ray irradiation (0,1,2,3,5,7 and 9Gy for chlnogenic assay or 1.5Gy for micronucleus assay) with various time intervals. Survival fractions of cells in both low dose-irradiated and unirrated groups were analyzed by clonogenic assay. Surviva fractions of low dose-irradiated in cell survival was maximum when low and high dose irradiation time interval was 4 hr. Frequencies of micronuclei which is an indicative of chromosome aberration were also enutained from survival fractions analyzed by clonogenic assay, maximum when low and high dose irradiation time interval was 4hr. Frequencies of micronuclei which is an indicative of chromosome aberration were also enumerated in both low dose-irradiated and unirradiated groups. In consiststent with the result obtained from survival fractions analyzed by clonogenic assay, maximum reduction in frquencies of micronuclei was observed when low dose radiation was given 4 hr prior to high response to subsequent high dose .gamma.-ray irradiation in human cervical carcinomal cells. Our data suggest that one of the possible mechanisms of adaptive response induced by low dose rediation is the increase in repair of DNA double strand breaks in low dose radiation-adapted cells.

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Genetic Diversity of Mulberry Dwarf Phytoplasma(MD) by SSCP Technique (SSCP기법에 의한 뽕나무오갈병 파이토플라스의 유전적 다형성 분석)

  • Han, Sangsub
    • Journal of Korean Society of Forest Science
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    • v.102 no.2
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    • pp.223-228
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    • 2013
  • Phytoplasmas were detected consistently in 42 mulberry cultivars showing dwarf disease using DNA analysis by amplification with phytoplasma universal primer pairs P1/P7 (about 1.8 kb and R16F2n/R2 (about 1.2 kb). The point mutation from 42 cultivars of mulberry tree was detected by single-strand conformation polymorphism (SSCP) analysis. The SSCP profiles were clearly observed from all of cultivars in 8% polyacrylamide gel, electrophoresizing for and running 8-15 hrs. at 150V, $10^{\circ}C$. The MD and JWB phytoplasma PCR products was mixed and electrophoresis was performed to detect their polymorphism. In this results, the SSCP profiles of all bands of MD and JWB were analyzed on single lane and were distinct in their each of band patterns. The SSCP analysis was possible to detect of 1.8 kb and 1.2 kb nucleotide size and near close band patterns were distinct by mix of two samples. Previously, it was only possible to detect of point mutation under 600 bp nucleotide sequence by SSCP analysis but this modification of SSCP technique was possible to detect clearly SSCP band patterns of about 1.8 kb and 1.2 kb nucleotides.

Antioxidant Activity and Protective Effects of Extracts from Chrysanthemum boreale on t-BHP Induced Oxidative Stress in Chang Cells (산국대 추출물의 항산화 활성 및 간세포 보호 효과)

  • Kim, Yon-Suk;Hwang, Jin-Woo;Park, Pyo-Jam;Jeong, Jae-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.1
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    • pp.60-66
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    • 2014
  • The aim of this study was to evaluate the antioxidant activity and protective effect of extracts from the stems and leaves of Chrysanthemum boreale (CBSL) on t-BHP induced oxidative stress in human liver cells (Chang cells). Antioxidant activities in the extracts were determined for various radical scavenging activities including ferric reducing antioxidant power, 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity, and oxygen radical absorbance capacity (ORAC). CBSL showed a very good scavenging effect of DPPH radical ($IC_{50}$ $0.009{\pm}0.002$ mg/mL), alkyl radical ($IC_{50}$ $0.004{\pm}0.001$ mg/mL), and hydroxyl radical ($IC_{50}$ $6.742{\pm}0.152$ mg/mL). CBSL also showed a strong antioxidant effect in the ORAC assay. In the MTT assay on human liver cells (Chang cells), the extracts showed protective effects by increasing cell viability, decreasing ROS, and restoring mitochondria membrane potential upon t-BHP induced oxidative stress. Our findings suggest that CBSL extracts are a potential therapeutic with protective antioxidant effects upon oxidative stress.

Protective Effects of Paeonia japonica against Radiation-induced Damage (방사선 장해에 대한 백작약의 방호효과)

  • Oh, Heon;Park, Hae-Ran;Jeong, Ill-Yun;Kim, Sung-Ho;Jo, Sung-Kee
    • Journal of Radiation Protection and Research
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    • v.27 no.3
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    • pp.181-188
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    • 2002
  • We investigated the effect of Paeonia japonica (PJ) on radiation-induced oxidative damage to macromolecules in vitro and in vivo. The PJ reduced the tail moment (TM) which was a marker of DNA strand break in single-cell gel electrophoresis (SCGE; comet assay) in the human peripheral blood lymphocytes. Lipid peroxidation in the liver of the ICR mouse, measured as malondialdehyde (MDA), was also reduced by PJ administration. Ethanol fraction of PJ was more effective than polysaccharide fraction of that on reduction of TM in SCGE and lipid peroxidation. Also, Their activities to scavenge DPPH radicals and hydroxyl radicals were observed in vitro, and the activities were due to its ethanol fraction. It is plausible that scavenging of flee radicals by PJ extract may have played an important role in providing the protection against the radiation-induced damage. These results indicated that Paeonia japonica might be a useful radioprotector, especially since it is a relatively nontoxic natural product.

Hair Strengthening Effect of Silane Coupling and Carbodiimide Chemistry (카르보디이미드 반응과 실란 커플링을 이용한 모발강화 효과)

  • Son, Seong Kil;Choi, Wonkyung;Lim, Byung Tack;Song, Sang-hun;Kang, Nae Kyu
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.44 no.2
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    • pp.133-139
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    • 2018
  • Chemically damaged hair is vulnerable to external stimuli in daily life due to the weakened physical properties of the hair strand itself. The purpose of this work was to determine whether chemical conjugation between hair keratin proteins restores tensile strength and thus results inpreventing further deterioration under repeated combing. A model damaged hair tress was produced by a typical perm-process. Then, it was internally crosslinked by the bifunctional crosslinker (3-aminopropyl)triethoxysilane (APTES), via both silane coupling and carbodiimide chemistry. Physical properties, including tensile strength, Young's modulus, and plateau stress, were measured to verify the effect of internal crosslinking, and the existence of crosslinking was verified by Fourier transform infrared (FT-IR) spectroscopy. The degrees of hair breakage and split ends were evaluated by repeated combing-drying tests. Physical properties of chemically damaged hair were restored by internal crosslinking. Successful crosslinking of APTES via both silane coupling and carbodiimide chemistry was verified by FT-IR spectra. Prevention of breakage and split ends after repeated combing with heat was observed. Human hair can be weakened by chemical damage including perm-processing, so restoring such properties is a major issue in the hair care industry. This work shows that internal crosslinking of damaged hair via chemical conjugation would be a potent method to restore the healthy hair.

Comparative Study on Human Risk by Ionizing Radiation and Pesticide as Biological Information about Environmental Disaster (환경재해에 관한 생물정보로서의 이온화 방사선과 살충제의 인체 위해성 비교 연구)

  • Kim, Jin-Kyu;Hyun, Soung-Hee
    • Journal of Radiation Protection and Research
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    • v.26 no.4
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    • pp.385-392
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    • 2001
  • Environmental risk factors such as ionizing radiations, heavy metals, and pesticides can cause environmental disasters when they exist in excess. The increases in use of ionizing radiation and agricultural pesticide are somewhat related to the possibility of the disaster. The risk of radiation and pesticide was evaluated by means of the single cell gel electrophoresis (SCGE) assay on the human blood lymphocytes. The lymphocytes were irradiated with $0{\sim}2.0Gy$ of $^{60}Co$ gamma ray. Another groups of lymphocytes were exposed to various concentrations of parathion. Significantly increased tail moment, which was a marker of DNA strand breaks in SCGE assay, showed a clear dose- or concentration-response relationship. Parathion of a recommended concentration for agricultural use ($1mg {\ell}^{-1}$ ) has a strong cytotoxic effect on lymphocytes, which is equivalent to damage induced by 0.1 Gy of ${\gamma}$-ray. Furthermore, $2mg{\ell}^{-1}$ of parathion can give rise to DNA damage equivalent to that induced by 0.25 Gy at which the radiation-induced damage can start to develop into clinical symptoms. The comparative results of this study can provide an experimental basis and biological information for the prevention of environmental disaster.

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Enhanced Sensitivity and Long-Term G2/M Arrest in Adriamycin-treated DNA-PK-null Cells are Unrelated to DNA Repair Defects (DNA-PK-null 세포주의 adriamycin 처리에 의한 G2/M 세포주기 변화)

  • Kim, Chung-Hee;Kim, Jong-Soo;Van Cuong, Dang;Kim, Na-Ri;Kim, Eui-Yong;Han, Jin
    • Journal of Life Science
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    • v.13 no.3
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    • pp.241-247
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    • 2003
  • While the DNA-protein kinase (DNA-PK) complex, comprised of DNA-PKcs and Ku80, is primary involved in the repair of DNA double-strand breaks, it is also believed to participate in additional cellular processes. Here, treatment of embryo fibroblasts (MEFs) derived from either wild-type (Wt) or DNA-PKcs-null (DNA-$PKcs^{-/-}$) mice with various stress inducing agents revealed that adriamycin was markedly more cytotoxic for $Ku80^{-/-}MEFs$ and led to their long-term accumulation in the $G_2$/M phase. This differential response was not due to differences in DNA repair, since adrimycin-triggered DNA damage was repaired with comparable efficiency in both Wt and $Ku80^{-/-}MEFs$, but was associated with differences in the expression of important cell cycle regulatory genes. Our results support the notion that Ku80-mediated cytoprotection and $G_2$/M-progression are not only dependent on the cell's DNA repair but also may reflect Ku80's influence on additional cellular processes such as gene expression.

An Analysis of Length and Time in the Elementary Mathematics Textbooks: Focused on the Instructional Components of Measurement and Key Competencies in Mathematics (길이와 시간에 관한 초등학교 수학 교과서 분석: 측정의 교수·학습 요소 및 수학 교과 역량을 중심으로)

  • Pang, JeongSuk;Kwon, MiSun;Kim, MinJeong;Choi, InYoung;SunWoo, Jin
    • School Mathematics
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    • v.18 no.2
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    • pp.301-322
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    • 2016
  • Even though measurement is an important strand of elementary mathematics education, there has been lack of research in this field. This study analyzed topics related to length and time in a series of mathematics textbooks aligned to 2007 or 2009 revised mathematics curriculum. The analysis was focused on three aspects: (a) overall instructional components of measurement, (b) instructional components specific to the topics of measurement, and (c) key competencies in mathematics. The results of this study showed that many topics dealing with length and time were represented with relation to real-life contexts or other subjects. The meanings of measurement terms and the necessity of calculation were well explained but other aspects still had room for improvement when it comes to the necessity of measurement units, appropriate choice of units, and use of students' common misconceptions. Another noticeable result was that problem solving, communication, and reasoning among key competencies in mathematics have been emphasized in the mathematics textbooks. Based on these results, this study provides textbook writers with implications on what to further consider in dealing with length and time.

Static Bending Strength Performances of Hybrid Laminated Woods Composed of Wood-Wood Based Boards (목재와 목질보드 복합적층재의 휨강도성능)

  • Park, Han-Min;Moon, Sung-Jae;Choi, Yoon-Eun;Park, Jung-Hwan;Byeon, Hee-Seop
    • Journal of the Korean Wood Science and Technology
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    • v.37 no.6
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    • pp.546-555
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    • 2009
  • To study an effective use of woods, three-ply hybrid laminated woods instead of crosslaminated woods which are composed of spruce in the face and three kinds of wood-based boards (MDF, PB, OSB) in the core were manufactured, and the effect of constitution elements for the core laminae on bending strength performances was investigated. Bending modulus of elasticity (MOE) of hybrid laminated woods had the highest values for the hybrid laminated wood types arranging OSB laminae in the core, and had the lowest values for those arranging MDF laminae in the core. These values were higher than those of various cross-laminated woods. The estimated bending MOEs of the hybrid laminated woods which were composed of perpendicular-direction lamina of spruce in the faces were similar to their measured values, regardless of wood-based boards in the core. However, those of the hybrid laminated woods which were composed of parallel-direction lamina of spruce in the faces had much higher values than those of their measured values, and it was necessary to revise the measured values. Bending modulus of rupture (MOR) of the hybird laminated woods had the highest value for those arranging OSB laminae in the core, and had the lowest values for those arranging PB laminae in the core unlike the bending MOE. By hybrid laminating, the anisotropy of bending strength performances was markedly decreased, and the differences of strength performances among wood-based boards were also considerably decreased.