• Title, Summary, Keyword: E-Science

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Expression of Human Papillomavirus Type 16, Prototype and Natural Variant E7 Proteins using Baculovirus Expression System

  • Han, Hee-Sung;Kee, Sun-Ho;Hwang, Soon-Bong;Kim, Hyung-Jun;Cho, Kyung-A;Kim, Yoon-Won;Cho, Min-Kee;Chang, Woo-Hyun
    • The Journal of Korean Society of Virology
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    • v.28 no.1
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    • pp.53-62
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    • 1998
  • Human papillomavirus (HPV) 16, E7 proteins derived from the prototype (Bac73) and natural variant (Bac101) E7 open reading frame were produced in Sf9 insect cells. The variant E7 gene occurred naturally by substitution mutation at the position of 88 nucleotide, resulting serine instead of asparagine. Using E7 specific monoclonal antibody (VD6), both E7 proteins were identified in recombinant baculovirus infected SF9 cells. Radiolabelling and immunoprecipitation analysis revealed that both E7 proteins were phosphoproteins. Immunostaining result showed that E7 proteins were mainly localized in the cytoplasm. Nuclear form of E7 proteins was also detected after a sequential fractionation procedure for removing chromatin structure. Considering that the VD6 recognition site in E7 protein is located within 10 amino acid at the N-terminus, this region appears to be blocked by the nuclear component. Western blot analysis revealed that nuclear form was more abundant than cytoplasmic E7 proteins. Time course immunostaining showed that the primary location of E7 protein was the nucleus and exported to the cytoplasm as proteins were accumulated. These events occurred similarly in both Bac73 and Bac101 infected Sf9 cells, suggesting that these two proteins may have similar biological functions.

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A Novel Mutant of Human Papillomavirus Type 18 E6E7 Fusion Gene and its Transforming Activity

  • Zhou, Zhi-Xiang;Zhao, Chen;Li, Qian-Qian;Zeng, Yi
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.17
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    • pp.7395-7399
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    • 2014
  • Background: Persistent human papillomavirus (HPV) infection, especially with high-risk types such as HPV16 and HPV18, has been identified as the primary cause of cervical cancer. E6 and E7 are the major onco-proteins of high-risk HPVs, which are consistently expressed in HPV infected tissues but absent in normal tissues and represent ideal therapeutic targets for immunotherapy of cervical cancer. Materials and Methods: In this study, the optimized fusion gene HPV18 E6E7 (HPV18 ofE6E7) was constructed according to genetic codon usage for human genes. At the same time, for safety future clinical application, a mutant of HPV18 ofE6E7 fusion gene was generated by site-directed mutagenesis at L52G for the E6 protein and C98G for the E7 protein. Results: HPV18-E6E7 mutant (HPV18 ofmE6E7) constructed in this work not only lost the transformation capability for NIH 3T3 cells and tumorigenicity in BALB/c nude mice, but also maintained very good stability and antigenicity. Conclusion: These results suggest that the mutant should undergo further study for application as a safe antigenspecific therapeutic vaccine for HPV18-associated tumors.

e-AIRS: Construction of an Aerodynamic Integrated Research System on the e-Science Infrastructure (e-AITS: e-Science 인프라 기반의 항공우주 공력통합연구 환경구축)

  • Kim, Jin-Ho;Yi, Jun-Sok;Ko, Soon-Heum;Ahn, Jae-Wan;Kim, Chong-Am;Kim, Yoon-Hee;Cho, Kum-Won
    • Journal of the Korean Society for Aeronautical & Space Sciences
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    • v.36 no.5
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    • pp.428-437
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    • 2008
  • e-AIRS, an abbreviation of ‘e-Aerospace Integrated Research System’, is a virtual organization designed to support the aerospace engineering processes in the e-Science environment. As the first step toward a virtual aerospace engineering organization, the e-AIRS intends to give a full support to aerodynamic research processes. Currently, the e-AIRS can handle both the computational and experimental aerodynamic researches on the e-Science infrastructure. In detail, users can conduct the full CFD(Computational Fluid Dynamics) research processes, request wind tunnel experiments, perform the comparative analysis between computational and experimental resultants and finally collaborate with other researchers using the web portal. The current paper will describe those functions and the internal architecture of the e-AIRS system.

Immunotherapeutic Effects of Dendritic Cells Pulsed with a Coden-optimized HPV 16 E6 and E7 Fusion Gene in Vivo and in Vitro

  • Zhou, Zhi-Xiang;Li, Dan;Guan, Shan-Shan;Zhao, Chen;Li, Ze-Lin;Zeng, Yi
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.9
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    • pp.3843-3847
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    • 2015
  • Background: Cervical cancer is the second most common cause of cancer related death of women. Persistent HPV infection, especially with high-risk types such as HPV16 and HPV18, has been identified to be the primary cause of cervical cancer. E6 and E7 are the major oncoproteins of high-risk HPVs, which are expressed exclusively in HPV infected tissues, and thereby represent ideal therapeutic targets for immunotherapy of cervical cancer. Materials and Methods: In this work, we used recombinant adenovirus expressing coden-optimized HPV16 E6 and E7 fusion protein (Ad-ofE6E7) to prime dendritic cells (DC-ofE6E7), to investigate the ability of primed DC vaccine in eliciting antitumor immunity in vitro and vivo. Results: Our results indicated that DC-ofE6E7 vaccine co-culturing with splenocytes could strongly induce a tumor-specific cytotoxic T lymphocyte (CTL) response and kill the TC-1 cells effectively in vitro. Moreover, DC-ofE6E7 vaccine induced protective immunity against the challenge of TC-1 cancer cells in vivo. Conclusions: The results suggested that the HPV16 ofE6E7 primed DC vaccine has potential application for cervical cancer immunotherapy.

Determinants of E-Government Assimilation in Indonesia: An Empirical Investigation Using a TOE Framework

  • Pudjianto, Boni;Zo, Hangjung;Ciganek, Andrew P.;Rho, Jae-Jeung
    • Asia pacific journal of information systems
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    • v.21 no.1
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    • pp.49-80
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    • 2011
  • E-government needs to be successfully implemented and assimilated into organizations to take advantage of its potential values and benefits for organizations. This study examines factors for e-government assimilation in Indonesia and employs the TOE (Technology-Organization-Environment) framework to develop a theoretical model to explain e-government assimilation. It also investigates how organizational type (central vs. local) plays a role in the assimilation of e-government. One hundred eighteen respondents from the central and local governments in Indonesia participated in the survey and an in-depth analysis based on partial least squares (PLS) was carried out. The results show that ICT infrastructure has the strongest significant relationship with e-government assimilation, Top management support, regulatory environment, ICT expertise, and competitive environment are also significant factors to explain e-government assimilation in Indonesia. Central and local governments Significantly differ in terms of e-government assimilation, so organizational type can be a moderator in the process of e-government assimilation. These findings present the efficacy of the proposed model for analyzing e-government assimilation and contribute additional insights for academia as well as practitioners and policy makers.

A Study of Scientifically Gifted High School Students' Perceptions on the Research and Education Program (R&E 프로그램에 대한 과학영재고등학생들의 인식 연구)

  • Kang, Seong-Joo;Kim, Hyun-Joo;Lee, Gil-Jae;Kwon, Young-Sik;Kim, Myung-Hee;Kim, Yeon-Suk;Kim, Yun-Hwa;Shin, Ho-Sim;Lim, Hee-Young;Ha, Ji-Hee
    • Journal of The Korean Association For Science Education
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    • v.29 no.6
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    • pp.626-638
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    • 2009
  • The purpose of this study was to investigate the perception of the scientifically gifted high school students to the research and education (R&E) program. 270 scientifically gifted high school students participating in R&E program were sampled and surveyed according to the R&E' purpose, the process of topic selection, the advantage, and disadvantage of R&E. 4 students among them were interviewed in depth. Students perceived the purposes and advantages of R&E as fostering the scientist' qualifications, providing the research opportunities, and improving the inquiry ability and the disadvantages of R&E as the time consumption in research, the participation as a sideline, and the difficulty of contents. The R&E program was mainly composed of a pre-determined topic, fixed procedures, and expert's settlements in the problem-emerging situation and was not cognitive but technical apprenticeship. In addition, students considered the participation of R&E as a chance of experience about the high techniques and the valuable instruments instead of the improvement on the creative problem-solving ability. The role of mentors in the R&E program is so important that it affects largely on the productive participation and the perceptions of science and scientists.

Environmental effects in the stellar populations of Compact Elliptical galaxies

  • Kim, Suk;Jeong, Hyunjin;Lee, Youngdae;Joo, Seok-Joo;Lee, Jaehyun;Sung, Eon-Chang
    • The Bulletin of The Korean Astronomical Society
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    • v.42 no.1
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    • pp.30.2-31
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    • 2017
  • Compact elliptical (cE) galaxies are in a rare class of stellar systems characterized by high stellar densities, small sizes, high velocity dispersion, and high metallicity corresponding to elliptical galaxies. cE galaxies have been observed around massive galaxies, so they could be formed under strong influences of tidal stripping and truncation. However, the recent discovery of isolated cE galaxies requires the need of new formation scenarios. We aim at finding cE galaxies in various environments using SDSS DR12, and studying stellar population of cEs as function of environments. Based on the typical properties of cE galaxies, we selected cE candidates by restricting that low-luminosity Mg > 19.5 mag, small sizes Re < 700 pc, and high velocity dispersions ${\sigma}$ > $60kms^{-1}$. Since effect radii of cE candidates are mostly smaller than the seeing size of SDSS photometry, we calculated the effective radius by fitting a Sersic profile. In addition, we assumed that host galaxies have brightness with Mr < -21 mag, and an environmental parameter is computed as distances between cE galaxies and host-galaxies. We found 112 cE galaxies at z < 0.05, which have high sersic indices (mean value is 5.2) similar to the typical massive elliptical galaxies. Mgb values of cE galaxies increase as the distances from the host galaxies decrease. Especially, for cEs close to the host galaxies (NcE; $D_{host}$ < 300 pc), the Mgb values are similar to those of massive elliptical galaxies, which is consistent with the previous studies. On the other hand, cE galaxies distant from the host galaxies (DcE; Dhost >300 pc) have lower Mgb values than the conventional cE. The Mgb values follow the ${\sigma}$-Mgb relation of elliptical galaxies, and are connected to its faint end. This can be explained as a result of different merger histories for differing environments. For example, NcE galaxies are formed by tidal stripping by massive galaxies as suggested by previous studies, but DcE galaxies could be linked with high-redshift spheroids (e.g. red nuggets) which have not evolved into present-day elliptical galaxies because of the environmental influences.

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Protective Immunity of Pichia pastoris-Expressed Recombinant Envelope Protein of Japanese Encephalitis Virus

  • Kwon, Woo-Taeg;Lee, Woo-Sik;Park, Pyo-Jam;Park, Tae-Kyu;Kang, Hyun
    • Journal of Microbiology and Biotechnology
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    • v.22 no.11
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    • pp.1580-1587
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    • 2012
  • Japanese encephalitis virus (JEV) envelope (E) protein holds great promise for use in the development of a recombinant vaccine. Purified recombinant E (rE) protein may be useful for numerous clinical applications; however, there are limitations in using the Escherichia coli expression system for producing high-quality rE protein. Therefore, in this study, the yeast expression system was used to generate the rE protein. For protein production using the yeast system, the full-length JEV E gene was cloned into Pichia pastoris. SDS-PAGE and immunoblotting analysis demonstrated that the rE protein had a molecular mass of 58 kDa and was glycosylated. The predicted size of the mature unmodified E protein is 53 kDa, suggesting that post-translational modifications resulted in the higher molecular mass. The rE protein was purified to greater than 95% purity using combined ammonium sulfate precipitation and a SP-Sepharose Fast Flow column. This purified rE protein was evaluated for immunogenicity and protective efficacy in mice. The survival rates of mice immunized with the rE protein were significantly increased over that of Hyphantria cunea nuclear polyhedrosis virus E protein (HcE). Our results indicate that the rE protein expressed in the P. pastoris expression system holds great promise for use in the development of a subunit vaccine against JEV.