• Title/Summary/Keyword: target life-time

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Glyceraldehyde-3-Phosphate Dehydrogenase, an Immunogenic Streptococcus equi ssp. zooepidemicus Adhesion Protein and Protective Antigen

  • Fu, Qiang;Wei, Zigong;Liu, Xiaohong;Xiao, Pingping;Lu, Zhaohui;Chen, Yaosheng
    • Journal of Microbiology and Biotechnology
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    • v.23 no.4
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    • pp.579-585
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    • 2013
  • Streptococcus equi ssp. zooepidemicus (Streptococcus zooepidemicus, SEZ) is an important pathogen associated with opportunistic infections of a wide range of species, including pigs and humans. The absence of a suitable vaccine makes it difficult to control SEZ infection. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) has been previously identified as an immunogenic protein using immunoproteomic techniques. In the present study, we confirmed that the sequence of GAPDH was highly conserved with other Streptococcus spp. The purified recombinant GAPDH could elicit a significant humoral antibody response in mice and confer significant protection against challenge with a lethal dose of SEZ. GAPDH could adhere to the Hep-2 cells, confirmed by flow cytometry, and inhibit adherence of SEZ to Hep-2 cells in an adherence inhibition assay. In addition, real-time PCR demonstrated that GAPDH was induced in vivo following infection of mice with SEZ. These suggest that GAPDH could play an important role in the pathogenesis of SEZ infection and could be a target for vaccination against SEZ.

Identification of Novel Universal Housekeeping Genes by Statistical Analysis of Microarray Data

  • Lee, Se-Ram;Jo, Min-Joung;Lee, Jung-Eun;Koh, Sang-Seok;Kim, So-Youn
    • BMB Reports
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    • v.40 no.2
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    • pp.226-231
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    • 2007
  • Housekeeping genes are widely used as internal controls in a variety of study types, including real time RT-PCR, microarrays, Northern analysis and RNase protection assays. However, even commonly used housekeeping genes may vary in stability depending on the cell type or disease being studied. Thus, it is necessary to identify additional housekeeping-type genes that show sample-independent stability. Here, we used statistical analysis to examine a large human microarray database, seeking genes that were stably expressed in various tissues, disease states and cell lines. We further selected genes that were expressed at different levels, because reference and target genes should be present in similar copy numbers to achieve reliable quantitative results. Real time RT-PCR amplification of three newly identified reference genes, CGI-119, CTBP1 and GOLGAl, alongside three well-known housekeeping genes, B2M, GAPD, and TUBB, confirmed that the newly identified genes were more stably expressed in individual samples with similar ranges. These results collectively suggest that statistical analysis of microarray data can be used to identify new candidate housekeeping genes showing consistent expression across tissues and diseases. Our analysis identified three novel candidate housekeeping genes (CGI-119, GOLGA1, and CTBP1) that could prove useful for normalization across a variety of RNA-based techniques.

Algorithm for Computational Age Dating of Nuclear Material for Nuclear Forensic Purposes

  • Park, Jaechan;Song, Jungho;Ju, Minsu;Chung, Jinyoung;Jeon, Taehoon;Kang, Changwoo;Woo, Seung Min
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.20 no.2
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    • pp.171-183
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    • 2022
  • The parent and daughter nuclides in a radioactive decay chain arrive at secular equilibrium once they have a large half-life difference. The characteristics of this equilibrium state can be used to estimate the production time of nuclear materials. In this study, a mathematical model and algorithm that can be applied to radio-chronometry using the radioactive equilibrium relationship were investigated, reviewed, and implemented. A Bateman equation that can analyze the decay of radioactive materials over time was used for the mathematical model. To obtain a differential-based solution of the Bateman equation, an algebraic numerical solution approach and two different matrix exponential functions (Moral and Levy) were implemented. The obtained result was compared with those of commonly used algorithms, such as the Chebyshev rational approximation method and WISE Uranium. The experimental analysis confirmed the similarity of the results. However, the Moral method led to an increasing calculation uncertainty once there was a branching decay, so this aspect must be improved. The time period corresponding to the production of nuclear materials or nuclear activity can be estimated using the proposed algorithm when uranium or its daughter nuclides are included in the target materials for nuclear forensics.

A proteomic approach reveals the differential protein expression in Drosophila melanogaster treated with red ginseng extract (Panax ginseng)

  • Liu, Qing-Xiu;Zhang, Wei;Wang, Jia;Hou, Wei;Wang, Ying-Ping
    • Journal of Ginseng Research
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    • v.42 no.3
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    • pp.343-351
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    • 2018
  • Background: Red ginseng is a popularly used traditional medicine with antiaging effects in Asian countries. The present study aimed to explore the changes in protein expression underlying the mechanisms of life span extension and antiaging caused by red ginseng extract (RGE) in Drosophila melanogaster. Methods: A proteomic approach of two-dimensional polyacrylamide gel electrophoresis (2-DE) was used to identify the differential abundance of possible target proteins of RGE in D. melanogaster. The reliability of the 2-DE results was confirmed via Western blotting to measure the expression levels of selected proteins. Proteins altered at the expression level after RGE treatment (1 mg/mL) were identified by matrix-assisted laser desorption/ionization-time of flight tandem mass spectrometry and by searching against the National Center for Biotechnology nonredundant and Uniprot protein databases. The differentially expressed proteins were analyzed using bioinformatics methods. Results: The average survival life span of D. melanogaster was significantly extended by 12.60% with RGE treatment (1 mg/mL) compared to untreated flies. This followed increased superoxide dismutase level and decreased methane dicarboxylic aldehyde content. Based on the searching strategy, 23 differentially expressed proteins were identified (16 up-regulated and 7 down-regulated) in the RGE-treated D. melanogaster. Transduction pathways were identified using the Kyoto Encyclopedia of Genes and Genomes database, and included the hippo and oxidative phosphorylation pathways that play important roles in life span extension and antiaging process of D. melanogaster. Conclusion: Treatment with RGE in D. melanogaster demonstrated that mechanisms of life span extension and antiaging are regulated by multiple factors and complicated signal pathways.

Ginsenoside compound K inhibits nuclear factor-kappa B by targeting Annexin A2

  • Wang, Yu-Shi;Zhu, Hongyan;Li, He;Li, Yang;Zhao, Bing;Jin, Ying-Hua
    • Journal of Ginseng Research
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    • v.43 no.3
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    • pp.452-459
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    • 2019
  • Background: Ginsenoside compound K(C-K), a major metabolite of ginsenoside, exhibits anticancer activity in various cancer cells and animal models. A cell signaling study has shown that C-K inhibited nuclear factor-kappa B ($NF-{\kappa}B$) pathway in human astroglial cells and liver cancer cells. However, the molecular targets of C-K and the initiating events were not elucidated. Methods: Interaction between C-K and Annexin A2 was determined by molecular docking and thermal shift assay. HepG2 cells were treated with C-K, followed by a luciferase reporter assay for $NF-{\kappa}B$, immunofluorescence imaging for the subcellular localization of Annexin A2 and $NF-{\kappa}B$ p50 subunit, coimmunoprecipitation of Annexin A2 and $NF-{\kappa}B$ p50 subunit, and both cell viability assay and plate clone formation assay to determine the cell viability. Results: Both molecular docking and thermal shift assay positively confirmed the interaction between Annexin A2 and C-K. This interaction prevented the interaction between Annexin A2 and $NF-{\kappa}B$ p50 subunit and their nuclear colocalization, which attenuated the activation of $NF-{\kappa}B$ and the expression of its downstream genes, followed by the activation of caspase 9 and 3. In addition, the overexpression of Annexin A2-K320A, a C-K binding-deficient mutant of Annexin A2, rendered cells to resist C-K treatment, indicating that C-K exerts its cytotoxic activity mainly by targeting Annexin A2. Conclusion: This study for the first time revealed a cellular target of C-K and the molecular mechanism for its anticancer activity.

Functional characterization and expression analysis of c-type and g-like-type lysozymes in yellowtail clownfish (Amphiprion clarkii)

  • Gaeun Kim;Hanchang Sohn;WKM Omeka;Chaehyeon Lim;Don Anushka Sandaruwan Elvitigala;Jehee Lee
    • Fisheries and Aquatic Sciences
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    • v.26 no.3
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    • pp.188-203
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    • 2023
  • Lysozymes are well-known antibacterial enzymes that mainly target the peptidoglycan layer of the bacterial cell wall. Animal lysozymes are mainly categorized as g-type, c-type, and i-type based on protein sequence and structural differences. In this study, c-type (AcLysC) and g-like-type (AcLysG-like) lysozymes from Amphiprion clarkii were characterized in silico via expressional and functional approaches. According to in silico analysis, open reading frames of AcLysC and AcLysG-like were 429 bp and 570 bp, respectively, encoding the corresponding polypeptide chains with 142 and 189 amino acids. Elevated expression levels of AcLysC and AcLysG-like were observed in the liver and the heart tissues, respectively, as evidenced by quantitative real-time polymerase chain reaction assays. AcLysC and AcLysG-like transcript levels were upregulated in gills, head kidney, and blood cells following experimental immune stimulation. Recombinant AcLysC exhibited potent lytic activity against Vibrio anguillarum, whereas recombinant AcLysG-like showed remarkable antibacterial activity against Vibrio harveyi and Streptococcus parauberis, which was further evidenced by scanning electron microscopic imaging of destructed bacterial cell walls. The findings of this study collectively suggest the potential roles of AcLysC and AcLysG-like in host immune defense.

Direction of Development and Alternatives in the Smart Citizens Party PR Strategy (스마트시민정당 PR전략의 발전방향과 대안)

  • Kim, Man-Ki
    • Journal of Digital Convergence
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    • v.9 no.1
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    • pp.189-200
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    • 2011
  • Social media environment has had a significant effect on 'power shift' over the world. Such change was due to the choice of smart people. Accordingly, the Smart Citizens Party PR strategies of $S^6M^2AR^3T^4$ should be formulated in accordance with times. Now, politicians or parties can gain a Specific, Simplicity, Sharing and Strong Synergy effect through Social Network Service (SNS). In addition, in order to embody Mighty, Measurable, and Achievable policies, policy-makers should formulate policies based on a Realistic basis, which can gain people's Reliability in every field Relevant to people's way of a happy life by giving them Time of equal opportunity and making them share the same vision and Target with Target segmentation for all sorts of people, Together. In this sense, twelve kinds of alternative words in The Smart Citizens Party PH strategies are suggested. The study suggests desirable direction of development and alternatives in regard to the trends of The Smart Citizens Party PR strategies. Alternatives that are given in this research paper will receive the people's choice because they are a dynamic force to execute 'mighty and powerful Korea'.

Development of EvaGreen Based Real-time PCR Assay for Detection and Quantification Toxic Dinoflagellate Pfiesteria Piscicida and Field Applications (유독 와편모조류 Pfiesteria Piscicida 탐지 및 정량 분석을 위한 EvaGreen 기반 Real-time PCR기법 개발과 현장 적용)

  • PARK, BUM SOO;JOO, JAE-HYOUNG;KIM, MYO-KYUNG;KIM, JOO-HWAN;KIM, JIN HO;BAEK, SEUNG HO;HAN, MYUNG-SOO
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.22 no.1
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    • pp.31-44
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    • 2017
  • Pfiesteria piscicida is one of heterotrophic dinoflagellate having toxic metaboliges, and it is difficult to detect and quantify this dinoflagellate via light microscope due to small size and morphological similarity with Pfiesteria-like dinoflagellate (PLD) species. Alternatively, we developed quantitative real-time PCR assay based on EvaGreen and determined field accessibility throughout the investigation of distribution in the entire Korean coastal waters and population dynamics in Shihwa Lake. The P. piscicida-specific primers based on internal transcribed spacer 1 (ITS 1) were designed and the specificity of primers was confirmed by PCR with other genomic DNAs which have genetic similarity with target species. Through real-time PCR assay, a standard curve which had a significant linear correlation between log cell number and $C_T$ value ($r^2{\geq}0.999$) and one informative melting peak ($88^{\circ}C$) were obtained. These results implies that developed real-time PCR can accurately detect and quantify P. piscicida. Throughout the field applications of real-time PCR assay, P. piscicida was distributed in western (Mokpo and Kimje) and easthern (Gangneng) Korean coastal water even though light microscopy failed to identify P. piscicida. In the investigation of population dynamics in Shihwa Lake, the density of P. piscicida was peaked in June, July and August 2007 at St. 1 where salinity (${\leq}15psu$) was lower than the other 2 sites. In this study, we successed to develop EvaGreen bassed real-time PCR for detection and quantification of P. piscicida in fields, so this developed assay will be useful for various ecological studies in the future.

Toward Generic, Immersive, and Collaborative Solutions to the Data Interoperability Problem which Target End-Users

  • Sanchez-Ruiz, Arturo;Umapathy, Karthikeyan;Hayes, Pat
    • Journal of Computing Science and Engineering
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    • v.3 no.2
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    • pp.127-141
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    • 2009
  • In this paper, we describe our vision of a "Just-in-time" initiative to solve the Data Interoperability Problem (a.k.a. INTEROP.) We provide an architectural overview of our initiative which draws upon existing technologies to develop an immersive and collaborative approach which aims at empowering data stakeholders (e.g., data producers and data consumers) with integrated tools to interact and collaborate with each other while directly manipulating visual representations of their data in an immersive environment (e.g., implemented via Second Life.) The semantics of these visual representations and the operations associated with the data are supported by ontologies defined using the Common Logic Framework (CL). Data operations gestured by the stakeholders, through their avatars, are translated to a variety of generated resources such as multi-language source code, visualizations, web pages, and web services. The generality of the approach is supported by a plug-in architecture which allows expert users to customize tasks such as data admission, data manipulation in the immersive world, and automatic generation of resources. This approach is designed with a mindset aimed at enabling stakeholders from diverse domains to exchange data and generate new knowledge.

Computer-Aided Drug Discovery in Plant Pathology

  • Shanmugam, Gnanendra;Jeon, Junhyun
    • The Plant Pathology Journal
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    • v.33 no.6
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    • pp.529-542
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    • 2017
  • Control of plant diseases is largely dependent on use of agrochemicals. However, there are widening gaps between our knowledge on plant diseases gained from genetic/mechanistic studies and rapid translation of the knowledge into target-oriented development of effective agrochemicals. Here we propose that the time is ripe for computer-aided drug discovery/design (CADD) in molecular plant pathology. CADD has played a pivotal role in development of medically important molecules over the last three decades. Now, explosive increase in information on genome sequences and three dimensional structures of biological molecules, in combination with advances in computational and informational technologies, opens up exciting possibilities for application of CADD in discovery and development of agrochemicals. In this review, we outline two categories of the drug discovery strategies: structure- and ligand-based CADD, and relevant computational approaches that are being employed in modern drug discovery. In order to help readers to dive into CADD, we explain concepts of homology modelling, molecular docking, virtual screening, and de novo ligand design in structure-based CADD, and pharmacophore modelling, ligand-based virtual screening, quantitative structure activity relationship modelling and de novo ligand design for ligand-based CADD. We also provide the important resources available to carry out CADD. Finally, we present a case study showing how CADD approach can be implemented in reality for identification of potent chemical compounds against the important plant pathogens, Pseudomonas syringae and Colletotrichum gloeosporioides.