• Title/Summary/Keyword: Rad54

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Cellular Dynamics of Rad51 and Rad54 in Response to Postreplicative Stress and DNA Damage in HeLa Cells

  • Choi, Eui-Hwan;Yoon, Seobin;Hahn, Yoonsoo;Kim, Keun P.
    • Molecules and Cells
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    • v.40 no.2
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    • pp.143-150
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    • 2017
  • Homologous recombination (HR) is necessary for maintenance of genomic integrity and prevention of various mutations in tumor suppressor genes and proto-oncogenes. Rad51 and Rad54 are key HR factors that cope with replication stress and DNA breaks in eukaryotes. Rad51 binds to single-stranded DNA (ssDNA) to form the presynaptic filament that promotes a homology search and DNA strand exchange, and Rad54 stimulates the strand-pairing function of Rad51. Here, we studied the molecular dynamics of Rad51 and Rad54 during the cell cycle of HeLa cells. These cells constitutively express Rad51 and Rad54 throughout the entire cell cycle, and the formation of foci immediately increased in response to various types of DNA damage and replication stress, except for caffeine, which suppressed the Rad51-dependent HR pathway. Depletion of Rad51 caused severe defects in response to postreplicative stress. Accordingly, HeLa cells were arrested at the G2-M transition although a small amount of Rad51 was steadily maintained in HeLa cells. Our results suggest that cell cycle progression and proliferation of HeLa cells can be tightly controlled by the abundance of HR proteins, which are essential for the rapid response to postreplicative stress and DNA damage stress.

Characterization of UV-damaged repair genes in cells

  • Choi, In-Soon
    • Journal of Life Science
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    • v.10 no.2
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    • pp.50-54
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    • 2000
  • The RAD4 gene of Saccharomyces cerevisiae is essential for the incision step of UV-induced excision repair. A yeast RAD4 gene has been previously isolated by functional complementation. In order to identify the RAD4 homologous gene from fungus Coprinus cinereus, we have constructed cosmid libraries from electrophoretically separated chromosomes of the C. cinereus. The 13 C. cinereus chromosomes were resolved by pulse-field gel electrophoresis, hybridized with S. cerevisiae RAD4 DNA, and then isolated homologous C. cinereus chromosome. Here, we report the cloning and characterization of fungus C. cinereus homolog of yeast RAD4 gene. Southern blot analysis confirmed that C. cinereus contains the sequence homologous DNA to RAD4 gene and this gene exists as a single copy in C. cinereus genome. When total RNA isolated from C. cinereus cells was hybridized with the 3.4 kb BglII DNA fragment of the S. cerevisiae RAD4 gene, a 2.5 kb of transcript was detected. The isolated gene encodes a protein of 810 amino acids.

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Stability of Human Centromeric Alphoid DNA Repeat during Propagation in Recombination-Deficient Yeast Strains (효모의 재조합 변이주를 이용한 인간 Centromeric Alphoid DNA Repeat의 안정성에 관한 연구)

  • Kim, Kwang-Sup;Shin, Young-Sun;Lee, Sang-Yeop;Ahn, Eun-Kyung;Do, Eun-Ju;Park, In-Ho;Leem, Sun-Hee;SunWoo, Yang-Il
    • Korean Journal of Microbiology
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    • v.43 no.4
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    • pp.243-249
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    • 2007
  • The centromere is a highly differentiated structure of the chromosome that fulfills a multitude of essential mitotic and meiotic functions. Alphoid DNA (${\alpha}$-satellite) is the most abundant family of repeated DNA found at the centromere of all human chromosomes, and chromosomes of primates in general. The most important parts in the development of Human Artificial Chromosomes (HACs), are the isolation and maintenance of stability of centromeric region. For isolation of this region, we could use the targeting hook with alphoid DNA repeat and cloned by Transformation-Associated Recombination (TAR) cloning technique in yeast Saccharomyces cerevisiae. The method includes rolling-circle amplification (RCA) of repeats in vitro to 5 kb-length and elongation of the RCA products by homologous recombination in yeast. Four types of $35\;kb{\sim}50\;kb$ of centromeric DNA repeat arrays (2, 4, 5, 6 mer) are used to examine the stability of repeats in homologous recombination mutant strains (rad51, rad52, and rad54). Following the transformation into wild type, rad51 and rad54 mutant strains, there were frequent changes in inserted size. A rad52 mutant strain showed extremely low transformation frequency, but increased stability of centromeric DNA repeat arrays at least 3 times higher than other strains. Based on these results, the incidence of large mutations could be reduced using a rad52 mutant strain in maintenance of centromeric DNA repeat arrays. This genetic method may use more general application in the maintenance of tandem repeats in construction of HAC.

ON WEAKLY 2-ABSORBING PRIMARY SUBMODULES OF MODULES OVER COMMUTATIVE RINGS

  • Darani, Ahmad Yousefian;Soheilnia, Fatemeh;Tekir, Unsal;Ulucak, Gulsen
    • Journal of the Korean Mathematical Society
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    • v.54 no.5
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    • pp.1505-1519
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    • 2017
  • Assume that M is an R-module where R is a commutative ring. A proper submodule N of M is called a weakly 2-absorbing primary submodule of M if $0{\neq}abm{\in}N$ for any $a,b{\in}R$ and $m{\in}M$, then $ab{\in}(N:M)$ or $am{\in}M-rad(N)$ or $bm{\in}M-rad(N)$. In this paper, we extended the concept of weakly 2-absorbing primary ideals of commutative rings to weakly 2-absorbing primary submodules of modules. Among many results, we show that if N is a weakly 2-absorbing primary submodule of M and it satisfies certain condition $0{\neq}I_1I_2K{\subseteq}N$ for some ideals $I_1$, $I_2$ of R and submodule K of M, then $I_1I_2{\subseteq}(N:M)$ or $I_1K{\subseteq}M-rad(N)$ or $I_2K{\subseteq}M-rad(N)$.

Reduction of Exposure dose in Cheat Roentgenography (흉부X선검사(胸部X線檢査)에 있어서 피폭선량(被曝線量)의 경감(輕減)에 대한 검토(檢討))

  • Huh, Joon;Kim, Chang-Kyun;Kang, Hong-Seok
    • Journal of radiological science and technology
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    • v.2 no.1
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    • pp.15-22
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    • 1979
  • Author made a experiment on the exposure dose with various intensifying screens in taking chest roentgenogram and obtained the results as follows; 1. Special speed type was the most sensitive intensifying screen, the r(gamma) value of this screen was distributed from 2.6 to 2.9. 2. The resolution activity of intensifying screen was inversely proportional to its sensitivity. If, the sensitivity and detail of the fine detail speed type intensifying screen at 100 KV were 100, those of the special speed type were 549 and 54.44 respectively. 3. If the exposure dose of the fine detail type intensifying screen was 1.0 at 60 KV, that of the special speed type intensifying screen was 0.1 at 80KV, and the skin dose of patient was as follows; it was 64.8 mRad at 60KV in mid speed type, 8.1 mRad at 80KV in super high speed type, and 7.2 mRad at 80KV in special speed type intensifying screen respectively.

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Comparison of Human Sodium/Iodide Symporter (hNIS) Gene Expressions between Lentiviral and Adenoviral Vectors in Rat Mesenchymal Stem Cells (렌티바이러스와 아데노바이러스를 통하여 쥐의 중간엽줄기세포에 사람 나트륨/옥소 공동수송체 유전자를 전달하였을 때의 발현성능 비교)

  • Park, So-Yeon;Kim, Sung-Jin;Lee, Won-Woo;Lee, Heui-Ran;Kim, Hyun-Joo;Chung, June-Key;Kim, Sang-Eun
    • Nuclear Medicine and Molecular Imaging
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    • v.42 no.5
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    • pp.394-400
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    • 2008
  • Purpose: Quantitative comparison of transgene expression within stem cells between lentivirus and adenovirus-mediated delivery systems has not been reported. Here, we evaluated the human sodium iodide symporter (hNIS) gene expression in rat mesenchymal stem cell (rMSC) transduced by lentivirus or adenovirus, and compared the hNIS expression quantitatively between the two delivery systems. Materials and Methods: Lentiviral-mediated hNIS expressing rMSC (lenti-hNIS-rMSC) was constructed by cloning hNIS gene into pLenti6/UbC/V5-DEST (Invitrogen) to obtain pLenti-hNIS, transducing rMSC with the pLenti-hNIS, and selecting with blasticidin for 3 weeks. Recombinant adenovirus expressing hNIS gene (Rad-hNIS) was produced by homologous recombination and transduction efficiency of Rad-hNIS into rMSC evaluated by Rad-GFP was $19.1{\pm}4.7%$, $54.0{\pm}6.4%$, $85.7{\pm}8.7%$, and $98.4{\pm}1.3%$ at MOI 1, 5, 20, and 100, respectively. The hNIS expressions in lenti-hNIS-rMSC or adeno-hNIS-rMSC were assessed by immunocytochemistry, western blot, and 1-125 uptake. Results: Immunocytochemistry and western blot analyses revealed that hNIS expressions in lenti-hNIS-rMSC were greater than those in adeno-hNIS-rMSC at MOI 20 but lower than at MOI 50. However in vitro 1-125 uptake test demonstrated that iodide uptake in lenti-hNIS-rMSC ($29,704{\pm}6,659\; picomole/10^6\;cells$) was greater than that in adeno-hNIS-rMSC at MOI 100 ($6,168{\pm}2,134\;picomole/10^6\;cells$). Conclusion: Despite lower amount of expressed protein, hNIS function in rMSC was greater by lentivirus than by adenovirus mediated expression. Stem cell tracking using hNIS as a reporter gene should be conducted in consideration of relative vector efficiency for transgene expression.

Physical Dosimetry in Radioactive Iodine Treatment in the Patients with Thyroid Cancer (갑상선암 환자에 대한 방사성옥소 치료시 물리적 선량 측정)

  • Kim, Myung-Seon;Jeong, Nae-In;Lee, Jai-Yong;Kim, Chong-Soon;Kim, Chong-Ho;Lee, Myung-Chul;Koh, Channg-Soon;Kim, Hee-Geun;Kang, Duck-Won;Song, Myung-Jae
    • The Korean Journal of Nuclear Medicine
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    • v.28 no.1
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    • pp.124-132
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    • 1994
  • Radioactive iodine has been widely used in patients with thyroid cancer combined with surgical treatment. However, due to individual variations in absorption and excretion and uptake by tumor tissue of radioactive iodine, there are differences in therapeutic effect and adverse effects even if the same doses are administrated. So this study compared the therapeutic effect and radiation hazard by measuring internal radiation dose. Of total 27 patients with well differentiated thyroid cancer who had been thyroidectomized, we administered radioactive iodine 100 mCi, 150 mCi, 200 mCi. According to BEL DOSIMETRY PROTO-COL, beta and gamma ray dose were estimated from a pelt of the logarithm of the percent of dose per liter of whole blood versus day, and percent dose retained versus day using somilogarithmic paper, respectively. 1) Physical dose to whole blood averaged $56.54{\pm}13.02$ rad in 100 mCi administered group, $76.83{\pm}19.97$ rad in 150 mCi administered group, $95.08{\pm}25.51$ rad in 200 mCi administered group and there has been a significant correlation among the groups. 2) Mean percent dose retained 48 hours later was 26.34%. 3) There was no significant correlation of physical dose between absence and presence of metastasis. 4) 17 of 19 patients who has been followed up with TSH and serum throglobulin, Thallium scan were successfully ablated by radioactive iodine. 5) Leukocyte, lymphocyte, neutrophil, platelet counts all deelined in 4.6 weeks and most of all were restored 3 months later. 6) There was no significant correlation between physical dosimetry and biologic dosimetry. Generally administered doses of radioactive iodine (100-200 mCi) to patients with thyroid cancer postoperatively had developed transient bone marrow suppression and minimal chromosomal aberration, but they were within safety dose to blood (200 rad). And there has been no significant differences in residual dose 48 hours later between Korean and western people.

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Analysis and Monitoring of Environmental Parameters in a Single-span Greenhouse during Strawberry Cultivation

  • Park, Minjung;Kang, Taegyeong;Yun, Sung-wook;Lim, Ryugap;Son, Jinkwan;Kang, Donghyeon
    • Journal of Environmental Science International
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    • v.30 no.11
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    • pp.907-914
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    • 2021
  • In this study, strawberry cultivation environment in a greenhouse located in Jeonju was monitored and internal environmental parameters were analyzed. Temperature, humidity, RAD, and PPF sensors were installed to monitor environmental conditions in the test greenhouse. Data were collected every 10 minutes during four winter months from sensors placed across the greenhouse to assess its permeability and environmental uniformity. Temperature and humidity inside the greenhouse were relatively uniform with negligible deviations among the center, south, and north; however, it was judged that further analysis of gradients of these parameters from the east to the west of the greenhouse would be needed. Both RAD (Total solar radiation) and PPF (Photosynthetic photon flux) had high values on the south and were low on the north and the reduction rate of these parameters was 54% and 61%, respectively, indicating that a significant amount of light could not be transmitted. This implied a significant decrease in the amount of light entering the greenhouse during winter. Therefore, it is concluded that environmental control devices and auxiliary lighting are needed to achieve uniform greenhouse environment for efficient strawberry cultivation.

An Experimental Research on Uniform Corrosion of Inconel 600 and 690 Tubing Material (Inconel 600 및 690 튜브 재질의 일반 부식에 관한 실험적 연구)

  • Yeom Yu-Sun;Hwang Jung-Lae;Jun In-Sub;Kim Soong-Pyung;Yoon Jang-Hee
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.4 no.2
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    • pp.103-116
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    • 2006
  • By executing corrosion experiment on Inconel 600, 690 used to material of S/G tube in domestic NPP, this paper show estimation of amount of product such as Co-58, Co-60, Cr-51, Mn-54, Fe-59 which are main exposure cause to the workers in NPP. Therefore, Making the 12 samples consisted of Inconel 600, 690, whole corrosion experiment was carried out for 60 days(each pH by 20 days). The conditions of those tests were similar or more harsh than actual conditions of domestic NPP. The Glow Discharge Spectrometer(GDS) was used for quantitative analysis of results. The results of using GDS, the Inconel 600 corrodes more than Inconel 690 at pH 7 and pH 9. However, it is observed that Inconel 690 corrodes more than Inconel 600 at pH 4. Those results is estimated that test sections had the effect of transient. The long terms of experiment is required to minimize and solve the problem.

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Seismic and progressive collapse assessment of SidePlate moment connection system

  • Faridmehr, Iman;Osman, Mohd Hanim;Tahir, Mahmood Bin Md.;Nejad, Ali Farokhi;Hodjati, Reza
    • Structural Engineering and Mechanics
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    • v.54 no.1
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    • pp.35-54
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    • 2015
  • The performance of a newly generated steel connection known as SidePlateTM moment connection for seismic loading and progressive collapse phenomenon has been investigated in this paper. The seismic evaluation portion of the study included a thorough study on of interstory drift angles and flexural strengths based on 2010 AISC Seismic Provisions while the acceptance criteria provided in UFC 4-023-03 guideline to resist progressive collapse must be satisfied by the rotational capacity of the connections. The results showed that the SidePlate moment connection was capable of attaining adequate rotational capacity and developing full inelastic capacity of the connecting beam. Moreover, the proposed connection demonstrated an exceptional performance for keeping away the plastic hinges from the connection and exceeding interstory drift angle of 0.06 rad with no fracture developments in beam flange groove-welded joints. The test results indicated that this type of connection had strength, stiffness and ductility to be categorized as a rigid, full-strength and ductile connection.