• Title/Summary/Keyword: Viral vector

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Pathological Lesions and Inducible Nitric Oxide Synthase Expressions in the Liver of Mice Experimentally Infected with Clonorchis sinensis

  • Yang, Qing-Li;Shen, Ji-Qing;Xue, Yan;Cheng, Xiao-Bing;Jiang, Zhi-Hua;Yang, Yi-Chao;Chen, Ying-Dan;Zhou, Xiao-Nong
    • Parasites, Hosts and Diseases
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    • v.53 no.6
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    • pp.777-783
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    • 2015
  • The nitric oxide (NO) formation and intrinsic nitrosation may be involved in the possible mechanisms of liver fluke-associated carcinogenesis. We still do not know much about the responses of inducible NO synthase (iNOS) induced by Clonorchis sinensis infection. This study was conducted to explore the pathological lesions and iNOS expressions in the liver of mice with different infection intensity levels of C. sinensis. Extensive periductal inflammatory cell infiltration, bile duct hyperplasia, and fibrosis were commonly observed during the infection. The different pathological responses in liver tissues strongly correlated with the infection intensity of C. sinensis. Massive acute spotty necrosis occurred in the liver parenchyma after a severe infection. The iNOS activity in liver tissues increased, and iNOS-expressing cells with morphological differences were observed after a moderate or severe infection. The iNOS-expressing cells in liver tissues had multiple origins.

Current trends of stem cell-mediated gene therapy (줄기 세포 분야의 유전자 치료 연구 동향)

  • Oh, Yu-Kyoung;Chung, Hyung-Min
    • Journal of Pharmaceutical Investigation
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    • v.32 no.2
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    • pp.65-72
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    • 2002
  • Recently, stem cell-mediated gene therapy is emerging as a novel therapeutic approach. For the successful gene modification of stem cells, the development of a suitable gene transfer technique needs to be preceded. This review focuses on the various gene transfer techniques based on nonviral and viral vectors, and physical methods. The advantages and disadvantages of each gene transfer method are compared, and the general properties of these vectors are discussed in relation to the gene transfer in stem cell research. This review also highlights the therapeutic application of stem cell-mediated gene therapy. The choice of gene transfer vectors may vary depending on the type of the stem cells and the target of stem cell therapy. Of various gene transfer methods, viral vector-based gene therapy has been emphasized due to the higher transfection efficiency. The current status and up-to-date findings of stem cell-mediated gene therapy are discussed in the viewpoint of the various targets of stem cell therapy such as the modification of stem cell potency, the acceleration of regeneration process and the formation of expressional organization.

Antibody-secreting macrophages generated using CpG-free plasmid eliminate tumor cells through antibody-dependent cellular phagocytosis

  • Cha, Eun Bi;Shin, Keun Koo;Seo, Jinho;Oh, Doo-Byoung
    • BMB Reports
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    • v.53 no.8
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    • pp.442-447
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    • 2020
  • The non-viral delivery of genes into macrophages, known as hard-to-transfect cells, is a challenge. In this study, the microporation of a CpG-free and small plasmid (pCGfd-GFP) showed high transfection efficiency, sustainable transgene expression, and good cell viability in the transfections of Raw 264.7 and primary bone marrow-derived macrophages. The non-viral method using the pCGfd vector encoding anti-EGFR single-chain Fv fused with Fc (scFv-Fc) generated the macrophages secreting anti-EGFR scFv-Fc. These macrophages effectively phagocytized tumor cells expressing EGFR through the antibody-dependent mechanism, as was proved by experiments using EGFR-knockout tumor cells. Finally, peri-tumoral injections of anti-EGFR scFv-Fc-secreting macrophages were shown to inhibit tumor growth in the xenograft mouse model.

A mutational anlaysis of the N-terminal protease of bovine viral diarrhea virus

  • Chon, Seung-ki
    • Korean Journal of Veterinary Research
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    • v.39 no.4
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    • pp.772-777
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    • 1999
  • The uncaped genomic RNA of bovine viral diarrhea virus (BVDV) initiates translation by recruitment of eukaryotic translation initiation factors at the internal ribosome entry site (IRES). N-terminal protease ($N^{pro}$) is the first translation product of the open reading frame (ORF). By using the vaccinia virus SP6 RNA polymerase transient expression system, we showed previously that deletion of $N^{pro}$ region reduced translation by 21%. To better understand the biological significance of $N^{pro}$ for translation, we carried out a mutational analysis of the $N^{pro}$ region of BVDV cloned in the intercistronic region of a bicistronic reporter plasmid. We constructed a bicistronic expression vector in which the entire 5 UTR and the mutated $N^{pro}$ region (${\Delta}386-901$, ${\Delta}415-901$ and ${\Delta}657-901$) was cloned between two reporter genes, chloramphenicol acetyltransferase (CAT) and luciferase (LUC). In vivo translation analyses showed that $N^{pro}$ region was dispensible for efficient translation. The results indicate that the $N^{pro}$ region is not essential for BVDV RNA translation and the 3' boundary of BVDV IRES is expanded into $N^{pro}$ region, suggesting that $N^{pro}$ may not play a major role in BVDV replication.

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Functional Implications in Apoptosis by Interferon Inducible Gene Product 1-8D, the Binding Protein to Adenovirus Preterminal Protein

  • Joung, In-Sil;Angeletti, Peter C.;Engler, Jeffrey A.
    • Journal of Microbiology
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    • v.41 no.4
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    • pp.295-299
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    • 2003
  • Adenovirus (Ad) precursor to the terminal protein (pTP) plays an essential roles in the viral DNA replication. Ad pTP serves as a primer for the synthesis of a new DNA strand during the initiation step of replication. In addition, Ad pTP forms organized spherical replication foci on the nuclear matrix (NM) and anchors the viral genome to the NM. Here we identified the interferon inducible gene product 1-8D (Inid) as a pTP binding protein by using a two-hybrid screen of a HeLa cDNA library. Of the clones obtained in this assay, nine were identical to the Inid, a 13-kDa polypeptide that shares homology with genes 1-8U and Leu-13/9-27, most of which have little known functions. The entire open reading frame (ORF) of Inid was cloned into the tetracycline inducible expression vector in order to determine the biological functions related with adenoviral infection. When Inid was introduced to the cells along with adenoviruses, fifty to sixty percent of Ad-infected cells expressing Inid had rounded morphology, which was suggestive of apoptosis. Results from the terminal deoxynucleotidyl transferase (TdT) and DNA fragmentation assays confirmed that Inid induces apoptosis in Ad-infected or in uninfected cells. The Inid binding to pTP may target the cell for apoptotic destruction as a host defense mechanism against the viral infection.

Identify the Risk Factors in Dengue Haemorrhagic Fever (DHF) using GIS

  • Nakhapakornc, Kanchana;Tripathi, Nitin;Nualchawee, Kaew;Kusanagi, Michiro;Pakpien, Preeda
    • Proceedings of the KSRS Conference
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    • 2003.11a
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    • pp.93-95
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    • 2003
  • Vector-borne diseases have been the most important worldwide health problem for many years and still represent a constant and serious risk to a large part of the world’s population. GIS and RS is used to evaluate and model the relationships between environmental factors/indicators and the incidences of viral diseases. The aim of the study is to identify the risk factors in Dengue Haemorrhagic Fever DHF) from the highest prevalence area and lowest prevalence area in Sukhothai province, Thailand using statistical, spatial and GIS Modeling. Results obtained in the study of the Dengue show that it is now possible to identify and localize precisely environmental indicators and factors of viral diseases.

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Development of Potato Virus Y Resistant Tobacco Plant by Transformation of the Untranslatable Viral Coat Protein Encoding cDNA (감자 바이러스 Y 비전이성 외피단백질 cDNA의 형질전환에 의한 바이러스 저항성 연초품종 개발)

  • 이청호;이영기;강신웅;박성원;김상석;박은경
    • Journal of the Korean Society of Tobacco Science
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    • v.19 no.2
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    • pp.117-123
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    • 1997
  • Viral coat protein (CP) encoding cDNA with artificial start and stop codons was synthesized by reverse-transcriptase polymerase chain reaction (RT-PCR) from the Korean isolate of potato virus Y-vein nectrosis strain (pVY-VN). To make PVY CP cDNA to untranslatable form, three stop codons were inserted near the start codon by "megaprimer-PCR" method. The untranslatable CP cDNA was subcloned to plant expression vector and transferred to N. tabacum cv. NC82 by Agrobacterium-mediated transformation. Highly resistant plants to PVY infection were screened, based on symptom development after mechanical virus inoculation. By genomic PCR and Southern blot analysis, one or more copies of the untranslatable CP gene were found in all transformants. From northern blot analysis, highly resistant transgenic lines had very low level of CP transcript but susceptible lines had high level, suggesting resistance to PVY infection should be related to RNA-mediated mechanism.mechanism.

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PVY Resistant Transgenic Potato Plants (cv Claustar) Expressing the Viral Coat Protein

  • Gargouri-Bouzid Radhia;Jaoua Leila;Mansour Riadh Ben;Hathat Yemna;Ayadi Malika;Ellouz Radhouane
    • Journal of Plant Biotechnology
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    • v.7 no.3
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    • pp.143-148
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    • 2005
  • The coat protein mediated resistance to potato virus Y is assessed here in transgenic potato plants (Solanum tuberosum L., cv Claustar). Therefore, the corresponding cDNA from tunisian isolate of the virus was cloned into Agrobacterium tumefaciens binary vector. The transgenic lines were subsequently analysed for the presence and expression of the transgene. The CP cDNA copy number was determined for kanamycin resistant plants. Three selected transgenic lines and their S1 progeny resulting from tuber germination showed a high protection level against the virus. These data appear to support the hypothesis that the virus resistance is mediated by the translated viral coat protein expressed in transgenic potato lines.

Studies on the Some Aspect of Small Brown Planthopper Transmission of Rice stripe tenuivirus (벼줄무늬잎마름병을 매개하는 애멸구의 전염생태)

  • Park, Jin-Woo;Lee, Min-Ho;Lee, Key-Woon
    • The Korean Journal of Pesticide Science
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    • v.15 no.4
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    • pp.490-494
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    • 2011
  • Rice stripe virus (RSV) has been the main viral disease of rice plant in western coastal region of Korea since 2000. The control of the vector insect, small brown planthopper (Laodelphax striatellus), is the most effective management method of the persistently-transmitted viral disease. Thus, ecological study between RSV and the vector insect was needed and investigated in order to make effective control plan, especially about study on the feeding and transmission of the virus by the vector insect. Each larval stage of vector insect differed in vector competence; larvae over 4th stage were shown as higher transmission after feeding on RSV-infected rice plant. These 4th and 5th larvae had higher transmission rates, 69.2% and 67.9% respectively, than 44.8% of the adult stage. The vector competence, however, was changed according to temperature; the highest transmission rate was 93.3% on $30^{\circ}C$ in comparison to 70.6% on $25^{\circ}C$ and 43.8% on $20^{\circ}C$.

Construction and Expression of an Eukaryotic Expression Vector Containing the IER3 Gene

  • Wang, Zhen;Yu, Hong-Sheng;Yao, Ru-Yong;Qiu, Wen-Sheng;Yue, Lu;Sui, Ai-Hua;Liu, Xiang-Ping;Liu, Shi-Hai
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.1
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    • pp.507-510
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    • 2013
  • Background: More and more research indicate that the immediately early response gene 3 (IER3) is involved inmany biological provesses, such as apoptosis and immunoreaction, as well as viral infection, tumorigenesis and tumour progression. Methods: Here we describe the construction of an eukaryotic expression vector containing IER3 gene and its expression in A549 cells as assessed through fluorescence microscopyand Western-blotting. Results: Fluorescence detection displayed that GFP in cytoplasm was high during 48 and 72 hours post-transfection. In addition, Western blotting showed significant increase in IER3 gene expression in the transfected cells compared with controls. Conclusion: The recombinate plasmid expression vector was constructed successfully, which may provide a basis for further exploration of function of IER3 in lung cancer.