• Title/Summary/Keyword: Single strand

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Identification of Six Single-Strand Initiation (ssi) Signals for Priming of DNA Replication in Various Plasmids

  • Jeong, Jin-Yong;Seo, Hak-Soo;Kim, Ho-Yeon;Cho, Moo-Je;Bahk, Jeong-Dong
    • BMB Reports
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    • v.28 no.4
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    • pp.336-341
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    • 1995
  • Using a mutant M13 phage derivative lacking a great part of the complementary strand synthesis origin, we identified six single-strand initiation (ssi) signals for DNA replication in pACYC184, pLG214, pGKV21, and pDPT270 plasmids, and named them $ssiA_{YC}$, $ssiA_{LG}$, $ssiB_{LG}$, $ssiA_{KV}$, $ssiA_{PT}$, and $ssiB_{PT}$, respectively. Two of them were from pDPT270, one from downstream the on of pACYC184, two from pLG214, one from upstream the plus origin of pGKV21. Introduction of these ssi signals into the deleted $ori_c$ site of a mutant filamentous M13 phage ($M13{\Delta}lac182$) resulted in the restoration of growth activity of this phage. These ssi signals were classified into a number of groups on the basis of sequence similarity. $ssiA_{YC}$ and $ssiA_{LG}$ show extensive sequence homology to the n'-site (primosome assembly sites) of ColE1, whereas $ssiB_{PT}$ is homologous to the n'-site of ${\Phi}X174$. $ssiA_{PT}$ belongs to G4-type ssi signals which require only dnaG primase and SSB protein for the priming of replication. In addition, possible biological roles of these ssi signals are discussed.

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Numerical study of mono-strand anchorage mechanism under service load

  • Marceau, D.;Fafard, M.;Bastien, J.
    • Structural Engineering and Mechanics
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    • v.18 no.4
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    • pp.475-491
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    • 2004
  • Anchorage devices play an important role in post-tensioned bridge structures since they must sustain heavy loads in order to permit the transfer of the prestressing force to the structure. In external prestressing, the situation is even more critical since the anchorage mechanisms, with the deviators, are the only links between the structure and the tendons throughout the service life of the structure. The behaviour of anchorage devise may be studied by using the finite element method. To do so, each component of the anchorage must be adequately represented in order to approximate the anchor mechanism as accurately as possible. In particular, the modelling of the jaw/tendon device may be carried out using the real geometry of these two components with an appropriate constitutive contact law or by replacing these components by a single equivalent. This paper presents the numerical study of a mono-strand anchorage device. The results of a comparison between two different representations of the jaw/tendon device, either as two distinct components or as a single equivalent, will be examined. In the double-component setup, the influence of the wedge configuration composing the jaw, and the influence of lubrication of the anchor, will be assessed.

Induction of Apoptosis and Single Strand Breaks by Extract of Pulsatilla Koreana (SB-31).

  • Kim, Sam-Yong;Kim, Hyun-Soo;Park, Sang-Jun;Kim, Jong-Suk;Park, Jee-Young;Yoon, Whan-Joong;Yoon, So-Hyun;Jo, Deog-Yeon
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.174-174
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    • 1996
  • Extract of Pulsatilla Koreana (SB-31) showed promising antitumor activity in vitro (J. Kor Cancer Asso 26:959-963, 1994). We studied the mechanism of cytotoxicity of SB-31. HL-60 cells were cocultivated with various concentrations of SB-31 for 5 hours. The DNAs from HL-60 cells exposed to SB-31 showed the ladder pattern typical of apoptosis. Effect of SB-31 on topoisomerase I activity was determined by slight modification of the method by E. Aflalo(1994). The pBR322 DNA showed dose-dependent increase of R-Form DNA upon incubation with SB-31. The topoisomerase Ⅰ-like activity (Increase of R-Form DNA) was accentuated with higher dose of SB-31. It is postulated that SB-31, which is a fermentation product of Pulsatilla koreana and which loses its activity when kept in ambient temperature for more than 96 hours, may contain topoisomerase Ⅰ-like activity and the enhanced excessive single strand breaks induced by 55-31 may result in apoptosis.

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Distinct Oxidative Damage of Biomolecules by Arrays of Metals Mobilized from Different Types of Airborne Particulate Matters: SRM1648, Fine (PM2.5), and Coarse (PM10) Fractions

  • Park, Yong Jin;Lim, Leejin;Song, Heesang
    • Environmental Engineering Research
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    • v.18 no.3
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    • pp.139-143
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    • 2013
  • This study was performed to examine the in vitro toxicities which are incurred due to the mobilization metals from standard reference material (SRM) 1648, fine ($PM_{2.5}$), and coarse ($PM_{10}$) particulate matter collected in Seoul metropolitan area. DNA single strand breaks of approximately 74% and 62% for $PM_{2.5}$ and for $PM_{10}$, respectively, were observed in the presence of chelator (EDTA or citrate)/reductant (ascorbate), as compared to the control by 2% without chelator or reductant. $PM_{2.5}$ induced about 40% more carbonyl formation with proteins in the presence of EDTA/ascorbate than $PM_{10}$. Therefore, more damage to biomolecules was incurred upon exposure to $PM_{2.5}$ than to $PM_{10}$. The treatment of a specific chelator, desferrioxamine, to the reaction mixture containing chelator plus reductant decreased the extent of damage to DNA to the level of the control, but did not substantially decrease the extent of damage to proteins. This suggests that different arrays of metals were involved in the oxidation of DNA and proteins.

Effects of Monosodium Glutamate on Unscheduled DNA Synthesis and DNA Single-Strand Breaks in Primary Cultures of Rat Hepatocytes (일차배양 간세포에서 Monosodium Glutamate에 의한 돌연변이 유발성의 검증)

  • 김동현;양규환
    • Environmental Mutagens and Carcinogens
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    • v.7 no.2
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    • pp.59-64
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    • 1987
  • Cytotoxic and genotoxic potential of monosodium glutamate (MSG) were evaluated in primary cultures of rat hepatocytes. When exposed to liver cell culture continuously for 24 hr, MSG did not show any cytotoxic effects up to 0.5% (w/v) level as determined by Tryphan Blue exclusion and lactic dehydrogenase release test. MSG also did not induce unscheduled DNA synthesis or DNA single-strand breaks in hepatocyte cultures up to 1% level. No synergistic effects of MSG were observed on aflatoxin B$_1$-induced DNA damage when 1% MSG was treated to liver cell culture along with aflatoxin B$_1$.

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Low-dose of Ultraviolet radiation-, Ethyl methanesulfonateor Bleomycin-lnduced Adaptive Response in Chinese hamster ovary Cells

  • Lee, Dong-Wook;Shin, Eun-Joo;Um, Kyung-Il
    • Environmental Mutagens and Carcinogens
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    • v.15 no.2
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    • pp.94-99
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    • 1995
  • The adaptive response and cross-adaptive response to sister chromatid exchanges (SCEs) and DNA single-strand breaks (SSBs) in Chinese hamster ovary (CHO)-K$_1$ cells treated with ultraviolet radiation (UV), ethyl methanesuffonate (EMS), or bleomycin (BLM) were investigated. Two assays were used in this study; SCEs and alkaline elution. The pretreatment with low conditioning dose of 2 mM EMS or 1 J/m$^2$ UV decreased the yield of SCEs induced by subsequent treatment with 8 mM EMS, 5 J/m$^2$ UV or 5 $\mu$g/ml BLM. And the pretreatment with low conditioning dose of 1 $\mu$g/ml BLM decreased the yield of SCEs induced by subsequent treatment with 5 $\mu$g/ml BLM or 5 J/m$^2$ UV. The rejoining of DNA SSBs in cells subsequently treated with 2 J/m$^2$ UV, 50 mM EMS or 400 $\mu$g/ml BLM is higher than that only treated with 2 J/m$^2$ UV, 50 mM EMS or 400 $\mu$g/ml BLM. These results suggest that there are the adaptive response and cross-adaptive response to SCEs, and is the adaptive response to the rejoining of DNA SSBs in CHO cells.

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Optimal Coil Configuration Design Methodology Using the Concept of Equivalent Magnetizing Current (등가자화전류를 이용한 최적코일형상 설계방법)

  • Kim, Woo-Chul;Kim, Min-Tae;Kim, Yoon-Young
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.31 no.1 s.256
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    • pp.43-49
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    • 2007
  • A new electric coil design methodology using the notion of topology optimization is developed. The specific design problem in consideration is to find optimal coil configuration that maximizes the Lorentz force under given magnetic field. Topology optimization is usually formulated using the finite element method, but the novel feature of this method is that no such partial differential equation solver is employed during the whole optimization process. The proposed methodology allows the determination of not only coil shape but also the number of coil turns which is not possible to determine by any existing topology optimization concept and to perform single coil strand identification algorithm. The specific applications are made in the design of two-dimensional fine-pattern focusing coils of an optical pickup actuator. In this method, the concept of equivalent magnetizing current is utilized to calculate the Lorentz force, and the optimal coil configuration is obtained without any initial layout. The method is capable of generating the location and shape of turns of coil. To confirm the effectiveness of the proposed method in optical pickup applications, design problems involving multipolar permanent magnets are considered.

Use of the Comet Assay to Assess DNA Damage in Hemocytes and Gill of Oyster(Crassostrea gigas) Exposed to Pyrene and Benzo(a)pyrene (Pyrene과 Benzo(a)pyrene에 노출된 굴의 혈구세포과 아가미 세포에서의 DNA손상 측정을 위한 Comet assay의 이용)

  • 김기범;배세진
    • Journal of Aquaculture
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    • v.16 no.3
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    • pp.196-201
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    • 2003
  • Sessile organisms such as the oyster Crassostrea gigas have been given much attention as a potential biomonitoring indicator to assess the impact of toxicants on aquatic organism. In this study, we exposed cells isolated from gill of oyster (Crassostrea gigas) to hydrogen peroxide in vitro. In addition oysters were in vivo exposed to pyrene and benzo(a)pyrene at various concentrations for 2 weeks. Comet assay was used to detect DNA single strand breaks and to investigate the application of this technique as a tool for aquatic biomonitoring. Hydrogen peroxide increased DNA single strand break with increasing concentration after 30 minutes exposure in vitro. Pyrene and benzo(a)pyrene caused DNA damage only at very high concentration (100 $\mu\textrm{g}$/L or 1000 $\mu\textrm{g}$/L) at two week exposure in vivo. DNA damage was relatively higher at hemocyte than at gill. It suggested that metabolized PAHs are transferred to hemolymph from digestive gland which have a relatively high enzyme activity, and attacked the DNA of hemocyte, while gill accumulated PAHs without degrading them to their metabolites due to low enzyme activity at gill. Both in vitro and in vivo exposure experiments showed that the comet assay is an effective tool on screening whether the organism are exposed to genotoxic contaminants.