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Deterimination of an Optimal Time Point for Analyzing Transcriptional Activity and Analysis of Transcripts of Avian Influenza Virus H9N2 in Cultured Cell (배양세포에서 Semi-quantitative RT-PCR에 의한 조류인플루엔자 H9N2의 전사활성 분석 최적 시기 결정 및 전사체 분석)

  • Na, Gi-Youn;Lee, Young-Min;Byun, Sung-June;Jeon, Ik-Soo;Park, Jong-Hyeon;Cho, In-Soo;Joo, Yi-Seok;Lee, Yun-Jung;Kwon, Jun-Hun;Koo, Yong-Bum
    • Korean Journal of Microbiology
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    • v.45 no.3
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    • pp.286-290
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    • 2009
  • The transcription of mRNA of avian influenza virus is regulated temporally during infection. Therefore, the measurement of transcript level in host cells should be performed before viral release from host cells because errors can occur in the analysis of the transcript levels if the viruses released from the infected cells re-infect cells. In this study, the timing of viral release was determined by measuring the level of viral RNA from viruses released from H9N2-infected chicken fibroblast cell line UMNSAH/DF-1 by semi-quantitative RT-PCR. The viral genomic RNA was isolated together with mouse total RNA which was added to the collected medium as carrier to monitor the viral RNA recovery and to use its GAPDH as an internal control for normalizing reverse transcription reaction as well as PCR reaction. It was found that viral release of H9N2 in the chicken fibroblast cell line UMNSAH/DF-1 took between 16 and 20 h after infection. We measured all 8 viral mRNA levels. Of the 8 transcripts, 7 species of viral mRNAs (each encoding HA, NA, PB1, PB2, NP, M, NS, respectively) except PA mRNA showed robust amplification, indicating these mRNA can be used as targets for amplification to measure transcript levels. These results altogether suggest that the method in this study can be used for screening antiviral materials against viral RNA polymerase as a therapeutic target.

Antioxidative Activity and Component Analysis of Fermented Melissa officinalis Extracts (레몬밤 발효추출물의 항산화 활성과 성분 분석)

  • Yang, Hee-Jung;Kim, Eun-Hee;Park, Jung-Ok;Kim, Jung-Eun;Park, Soo-Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.35 no.1
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    • pp.47-55
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    • 2009
  • In this study, the antioxidative effects, inhibitory effects on tyrosinase, and component analysis of fermented Melissa officinalis extracts were investigated. The ethyl acetate fraction of fermented extract ($8.38{\mu}g/mL$) showed the most prominent the free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities ($FSC_{50}$) of extract/fractions of M. officinalis. Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of some M. officinalis extracts on ROS generated in $Fe^{3+}$-EDTA/$H_{2}O_{2}$ system were investigated using the luminol-dependent chemiluminescence assay. The ethyl acetate fraction of fermented extract ($0.63{\mu}g/mL$) showed the most prominent ROS scavenging activity. The protective effects of extract/fractions of M. officinalis on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The M. officinalis extracts suppressed photohemolysis in a concentration dependent manner ($5\;{\sim}\;75{\mu}g/mL$). The inhibitory effect of M. officinalis extracts on tyrosinase was investigated to assess their whitening efficacy. Inhibitory effects ($IC_{50}$) on tyrosinase of some M. officinalis extracts was 50 % ethanol extract ($365{\mu}g/mL$) < ethyl acetate fraction of fermented extract ($122.43{\mu}g/mL$) < ethylacetate fraction ($94.8{\mu}g/mL$). Fractions of ethyl acetate both from ordinary and fermented M. officinalis extracts showed 2 band in TLC and 2 peak in HPLC (330 nm). In HPLC chromatogram of ethyl acetate fraction, peak 1 (51.64 %) and peak 2 (48.36 %) were identified as caffeic acid and rosmarinic acid in the order of elution time. Also, in HPLC chromatogram of ethyl acetate fraction of fermented extract, peak 1 (4.13 %) and peak 2 (95.87 %) were identified as caffeic acid and rosmarinic acid in the order of elution time. These results indicate that the component and content of ordinary and fermented extracts of M. officinalis are different. And the extract of M. officinalis can be used as an antioxidant.

Theoretical Research for Unmanned Aircraft Electromagnetic Survey: Electromagnetic Field Calculation and Analysis by Arbitrary Shaped Transmitter-Loop (무인 항공 전자탐사 이론 연구: 임의 모양의 송신루프에 의한 전자기장 반응 계산 및 분석)

  • Bang, Minkyu;Oh, Seokmin;Seol, Soon Jee;Lee, Ki Ha;Cho, Seong-Jun
    • Geophysics and Geophysical Exploration
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    • v.21 no.3
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    • pp.150-161
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    • 2018
  • Recently, unmanned aircraft EM (electromagnetic) survey based on ICT (Information and Communication Technology) has been widely utilized because of the efficiency in regional survey. We performed the theoretical study on the unmanned airship EM system developed by KIGAM (Korea Institute of Geoscience and Mineral resources) as part of the practical application of unmanned aircraft EM survey. Since this system has different configurations of transmitting and receiving loops compared to the conventional aircraft EM systems, a new technique is required for the appropriate interpretation of measured responses. Therefore, we proposed a method to calculate the EM field for the arbitrary shaped transmitter and verified its validity through the comparison with analytic solution for circular loop. In addition, to simulate the magnetic responses by three-dimensionally (3D) distributed anomalies, we have adapted our algorithm to 3D frequency-domain EM modeling algorithm based on the edge-FEM (finite element method). Though the analysis on magnetic field responses from a subsurface anomaly, it was found that the response decreases as the depth of the anomaly increases or the flight altitude increases. Also, it was confirmed that the response became smaller as the resistivity of the anomaly increases. However, a nonlinear trend of the out-of-phase component is shown depending on the depth of the anomaly and the used frequency, that makes it difficult to apply simple analysis based on the mapping of the magnitude of the responses and can cause the non-uniqueness problem in calculating the apparent resistivity. Thus, it is a prerequisite to analyze the appropriate frequency band and flight altitude considering the purpose of the survey and the site conditions when conducting a survey using the unmanned aircraft EM system.

Antioxidant Activity and Component Analysis of Fermented Lavandula angustifolia Extracts (라벤더 발효추출물의 항산화 활성과 성분 분석)

  • Park, Soo-Nam;Ahn, You-Jin;Won, Bo-Ryoung;Kang, Myung-Kyu;Kim, Jai-Hyun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.35 no.2
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    • pp.125-134
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    • 2009
  • In this study, the antioxidative effects, inhibitory effects on tyrosinase, and component of non-fermented and fermented Lavandula angustifolia extracts were investigated. The ethyl acetate fraction of fermented extract (5.95 ${\mu}g/mL$) showed the most prominent the free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of L. angustifolia extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The ethyl acetate fraction of fermented extract (1.45 ${\mu}g/mL$) showed the most prominent ROS scavenging activity. The protective effects of extract/fractions of L. angustifolia on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The L. angustifolia extracts suppressed photohemolysis in a concentration dependent manner (1 ${\sim}$ 50 ${\mu}g/mL$). The inhibitory effect of L. angustifolia extracts on tyrosinase was investigated to assess their whitening efficacy. Inhibitory effects ($IC_{50}$) on tyrosinase were determined with ethyl acetate fraction of L. angustifolia extract (144.80 ${\mu}g/mL$) and ethyl acetate fraction of fermented extract (122.40 ${\mu}g/mL$). Fractions of ethyl acetate and fermented extracts showed both 3 band in TLC and 3 peaks, 2 peaks in HPLC (340 nm), respectively. In each chromatography, fractions of ethyl acetate both from non-fermented and fermented L. angusfifolia have rosmarinic acid in common. These results indicate that the component and content of non-fermented and fermented extracts of L. angustifolia are different. Both of the extract of L. angustifolia can be used as an antioxidant.

The Mechanisms of Resistance to TNF in TNF-Sensitive Cancer Cells Transfected with TNF-$\alpha$ Gene Using Retroviral Vector (암세포에서 Retroviral Vector를 이용한 종양괴사인자 유전자 이입후 획득된 종양괴사인자 내성의 기전)

  • Lee, Hyuk-Pyo;Oh, Yeon-Mok;Yoo, Chul-Gyu;Kim, Young-Whan;Shim, Young-Soo;Han, Sung-Koo
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.3
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    • pp.547-558
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    • 1997
  • Background : Tumor necrosis factor(TNF) has been considered as an important candidate for cancer gene therapy based on its potent anti-tumor activity. However, since the efficiency of current techniques of gene transfer is not satisfactory, the majorities of current protocols is aiming the in vitro gene transfer to cancer cells and re-introducing genetically modified cancer cells to hoot. In previous study, it was shown that TNF-sensitive cancer cells transfected with TNF-$\alpha$ cDNA would become highly resistant to TNF. Understanding the mechanisms of TNF-resistance in TNF-$\alpha$ gene transfected cancer cells would be an important step for improving the efficacy of cancer gene therapy as well as for better understandings of tumor biology. This study was designed to evaluate the role of new protective protein synthesis in the acquired resistance to TNF of TNF-$\alpha$ gene transfected cancer cells. Method : We transfected TNF-$\alpha$ c-DNA to WEHI164, a murine fibrosarcoma cell line, using retroviral vector(pLT12SN(TNF)) and confirm the expression of TNF with PCR, ELISA, MIT assay. Then we determined the TNF resistance of TNF gene transfected cells(WEHI164-TNF) and the changes of TNF sensitivities after treatments with actinomycin D(transcription inhibitor) and cycloheximide ( translation inhibitor). Results : WEHI164 which was sensitive to TNF became resistant to TNF after being transfected with TNF-$\alpha$ gene and the resistance to TNF was partially reversed after treatment with actinomycin D, but not with cycloheximide. Conclusion : The acquired resistance to TNF after TNF-$\alpha$ gene transfection may be associated with synthesis of some protective proteins.

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Studies on Seed Production of Saddleback Clownfish, Amphiprion polymnus 1) Spawning, Egg Development and Larvae Culture (Saddleback clownfish, Amphiprion polymnus의 종묘생산에 관한 연구 1) 산란과 난 발생 및 자치어 사육)

  • Yoon, Young-Seock;Rho, Sum;Choi, Young-Ung;Kim, Jong-Su;Lee, Young-Don
    • Journal of Aquaculture
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    • v.18 no.2
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    • pp.107-114
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    • 2005
  • Clownfish are important and very popular fish in the ornamental aquarium industry. Demand for the fish is increasing dramatically. The present study was conducted to verify methods of broodstock management, patterns of spawning, rates of egg hatching and estimates of larval growth fur the saddleback clownfish, Amphiprion polymnus. Spawning occurred 8 times between August 2002 to June 2004 with 2 females and 1 male participating. Fertilized eggs were separated by an adhesive matrix and were oval in shape. The eggs were $2.46{\pm}0.13mm$ in size as measured along the longest axis. The percentage of fertilized eggs was 96.7%. Hatching was observed seven days post-spawning and hatching rate was 85.5%. The sizes of the newly-hatched larvae were $4.58{\pm}0.21mm$ TL (total length). Larvae had an open mouth and anus, and an oval yolk sac. At the 1 st day after hatching, the sizes of the larvae were $4.90{\pm}0.35mm$ TL. The larvae began to eat rotifers after complete yolk absorption. On the 5th day post-hatch, larvae were $5.88{\pm}0.31mm$ TL with complete fins and the survival rate was 48.6%. At 8 days after hatching, a band began to appear on head and back of the larvae indicating the beginning of metamorphosis. Metamorphosis was completed at an average TL of $15.00{\pm}2.12mm$ on the 23rd day after hatching. By the 45th day after hatching, juveniles averaged $22.76{\pm}3.22mm$ TL and survival rate was 28.4%.

Synthesis and Characterization of Layered Copper Hydroxides in Highly Concentrated Solution (고농도 용액에서 Layered Copper Hydroxides의 합성 및 특성)

  • Nam, Dae-Hyean;Choi, Choong-Lyeal;Kim, Kwang-Seop;Seo, Young-Jin;Park, Man
    • Korean Journal of Soil Science and Fertilizer
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    • v.43 no.6
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    • pp.872-879
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    • 2010
  • Layered copper hydroxides [LCHs, $Cu_2(OH)_3{\cdot}NO_3$] has the agricultural potentials as a fungicide because of its high crystallinity, excellent anion exchange capacity, and its regular layered particle size. The study, for the first time, has synthesized LCHs in highly concentrated solution and evaluated its physicochemical properties including the crystallinity and suspension stability. Optimal synthetic condition of LCHs was determined by crystallinity and stability of suspension as follow; 1) concentrations of $Cu(NO_3)_2$ and NaOH solutions were 3.0 M respectively, 2) reaction temperature and solution pH were $25^{\circ}C$ and 6.0, respectively, and 3) aging time after reaction was 2hr. Crystallinity of LCHs enhanced with increase in pH up to 9.0. Whereas, stability of suspension was decrease by increase in crystal size. Especially, increase in reaction temperature decreased stability of suspension. XRD patterns and SEM images exhibited that LCHs had regular layered particle size with 0.2~0.8 ${\mu}m$ and high crystallinity in optimal synthetic condition. The particle size was increased with increase in reaction temperature and pH. These results showed that LCHs synthesized in highly concentrated solution exhibited high stability of suspension as well as high crystallinity suitable to their potential as a fungicide.

Studies on the Electrochemical Behavior of Heavy Lanthanide Ions and the Synthesis, Characterization of Heavy Metal Chelate Complexes(II). Synthesis and Characterization of Eight Coordinate Tungsten(IV) and Cerium(IV) Chelate Complex (무거운 란탄이온의 전기화학적 거동 및 중금속이온의 킬레이트형 착물의 합성 및 특성에 관한 연구(제2보). 8배위 텅스텐(IV)과 세륨(IV)의 킬레이트형 착물의 합성 및 특성)

  • Kang, Sam Woo;Chang, Choo Wan;Suh, Moo Yul;Lee, Doo Youn;Choi, Won Jong
    • Analytical Science and Technology
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    • v.5 no.1
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    • pp.41-49
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    • 1992
  • An attempt was made to prepare two series of tetrakis eight-coordinate tungsten(IV) and cerium(IV) complexes containing the 5,7-dichloro-8-quinolinol(N:${\pi}$-acceptor atom, O:${\pi}$-donor atom) ligand. Tetrakis eight-coordinate tungsten(IV) complex of 2-mercaptopyrimidine(N:${\pi}$-acceptor atom, S:${\pi}$-donor atom) ligand have also been prepared. And the new series of mixed-ligand eight-coordinate tungsten(IV) complexes containing bidentate ligands 5,7-dichloro-8-quinolinol and 2-mercaptopyrimidine have been prepared, isolated by TLC and characterized. $W(dcq)_4$, $W(dcq)_3(mpd)_1$, $W(dcq)_2(mpd)_2$, $W(dcq)_1W(dcq)_3$ and $W(mpd)_4$ complexes of MLCT absorption band appeared to 710nm, 680nm, 625nm, 581nm, and 571nm(${\varepsilon}\;max={\sim}>{\times}10^4$) on low-energy respectively. The specific absorption wave length of $Ce(dcq)_4$ is appeared 520nm(${\varepsilon}\;max={\sim}>{\times}10^4$). The Chemical shift values by proton of coordinated position appeared to $W(dcq)_4$ [$H_2:8.9ppm$]; $W(dcq)_3(mpd)_1$ [$H_2:9.3$,$H_6:9.2ppm$]; $W(dcq)_2(mpd)_2$ [$H_2:9.7$,$H_6:8.95ppm$]; $W(dcq)_1(mpd)_3$ [$H_2:9.8$,$H_6:9.4ppm$]; $W(mpd)_4$ [$H_6:8.8ppm$]; $Ce(dcq)_4$ [$H_2:9.3ppm$] with $^1H$-NMR. The inertness of mixed-ligand eight coordinate tungsten(IV) complexes have been investigated by UV-Vis. spectroscopic method in dimethylsulfoxide at $90^{\circ}C$. The inertness of $W(dcq)_n(mpd)_{4-n}$ complexes showed the following order, $W(dcq)_3(mpd)_1;k_{obs.}=3.8{\times}10^{-6}$ > $W(mpd)_4;k_{obs.}=6.0{\times}10^{-6}$ > $W(dcq)_4;k_{obs.}=6.4{\times}10^{-6}$ > $W(dcq)_2(mpd)_2;k_{obs.}=7.0{\times}10^{-6}$ > $W(dcq)_1(mpd)_3;k_{obs.}=1.7{\times}10^{-5}$, which showed the inertness until 16days, 10days, 9days, 8days, and 4days. The $W(mpd)_4$ is very inert as $k_{obs.}=3.6{\times}10^{-6}$(16days) in xylene at $90^{\circ}C$ and $k_{obs.}=6.0{\times}10^{-6}$(10days) in DMSO at $90^{\circ}C$.

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Construction of cDNA Library for Using Virus-induced Gene Silencing (VIGS) Vector with the Sweetpotato Whitefly, Bemisia tabaci(Hemiptera: Aleyrodidae) (담배가루이(Bemisia tabaci, Aleyrodidae, Hemiptera)에서 Virus-induced Gene Silencing (VIGS) Vector를 이용하기 위한 cDNA Library 제작)

  • Ko, Na Yeon;Lim, Hyoun Sub;Yu, Yong Man;Youn, Young Nam
    • Korean journal of applied entomology
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    • v.54 no.2
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    • pp.91-97
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    • 2015
  • The sweetpotato whitefly, Bemisia tabaci, is the major insect pest that transmitted over 100 plant viruses including tomato yellow leaf curl virus (TYLCV) of tomato plant as virus vector in the world. In this study, cDNA library of whitefly was constructed using Gateway system for selecting target gene in order to control of B. tabaci using virus-induced gene silencing (VIGS) vector with RNAi. First of all, when using oligo d(T) rimer, the calculated titer of cDNA library was confirmed with $1.4{\times}10^4$ clones and average insert sizes was confirmed with 1 kb. However, insert size was very big for construction of cDNA. Otherwise, when using attB-N25 random primer and sonication for 6 sec, the calculated titer of cDNA library was confirmed with $1.04{\times}10^5$ clones. But mostly insert band wasn't identified on the electrophoresis, because it seemed that insert size is too small (${\leq}100bp$), also the size of identified insert was somewhat big. Finally, when using oligo d(T) primer and sonication for 1 sec, cDNA insert of whitefly was appropriated for VIGS with 300-600 bp. However, cDNA sequence included a poly A and titer was very low to $5.2{\times}10^2$ clones. It was supposed that heat shock transformation was used instead of electro-transformation. It is considered that when constructing cDNA library for using VIGS vector, (1) random primer should be used for First strand cDNA synthesis in order to remove poly A and (2) sonication for 1 sec should be performed in order to get appropriated insert size and (3) electro-transformation should be performed in order to improve transformation efficiency.

Change Analysis of Aboveground Forest Carbon Stocks According to the Land Cover Change Using Multi-Temporal Landsat TM Images and Machine Learning Algorithms (다시기 Landsat TM 영상과 기계학습을 이용한 토지피복변화에 따른 산림탄소저장량 변화 분석)

  • LEE, Jung-Hee;IM, Jung-Ho;KIM, Kyoung-Min;HEO, Joon
    • Journal of the Korean Association of Geographic Information Studies
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    • v.18 no.4
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    • pp.81-99
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    • 2015
  • The acceleration of global warming has required better understanding of carbon cycles over local and regional areas such as the Korean peninsula. Since forests serve as a carbon sink, which stores a large amount of terrestrial carbon, there has been a demand to accurately estimate such forest carbon sequestration. In Korea, the National Forest Inventory(NFI) has been used to estimate the forest carbon stocks based on the amount of growing stocks per hectare measured at sampled location. However, as such data are based on point(i.e., plot) measurements, it is difficult to identify spatial distribution of forest carbon stocks. This study focuses on urban areas, which have limited number of NFI samples and have shown rapid land cover change, to estimate grid-based forest carbon stocks based on UNFCCC Approach 3 and Tier 3. Land cover change and forest carbon stocks were estimated using Landsat 5 TM data acquired in 1991, 1992, 2010, and 2011, high resolution airborne images, and the 3rd, 5th~6th NFI data. Machine learning techniques(i.e., random forest and support vector machines/regression) were used for land cover change classification and forest carbon stock estimation. Forest carbon stocks were estimated using reflectance, band ratios, vegetation indices, and topographical indices. Results showed that 33.23tonC/ha of carbon was sequestrated on the unchanged forest areas between 1991 and 2010, while 36.83 tonC/ha of carbon was sequestrated on the areas changed from other land-use types to forests. A total of 7.35 tonC/ha of carbon was released on the areas changed from forests to other land-use types. This study was a good chance to understand the quantitative forest carbon stock change according to the land cover change. Moreover the result of this study can contribute to the effective forest management.