• Title/Summary/Keyword: recombinant virus

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Feline Herpes Virus-1 Associated Facial and Perianal Dermatitis in a Cat (고양이 헤르페스 바이러스-1 감염과 관련된 고양이의 안면 및 회음부 피부염)

  • Yoon, Ji-Seon;Yabuzoe, Astushi;Sekiguchi, Maiko;Park, Jinho;Iwasaki, Toshiroh;Nishifuji, Koji
    • Journal of Veterinary Clinics
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    • v.30 no.3
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    • pp.210-213
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    • 2013
  • We herein describe a feline case of facial dermatitis whose histopathological features resembled to those of FHV-associated ulcerative dermatitis. A 3-year-old, intact male domestic short-haired cat was presented with 2-years history of pruritic dermatitis that initially appeared on periocular area and extended toward the entire face. The cat had ocular discharge and conjunctivitis from 2-month of age. Clinically, skin lesions were characterized as erythema, erosions and ulcers covered with crusts on the facial and perianal area. Histopathologically, the facial lesion was characterized as interface dermatitis with hydropic degeneration at the basal layer, and single cell necrosis of keratinocytes. In addition, the epidermal and dermal necrosis infiltrated with eosinophils, and intranuclear inclusion bodies in keratinocytes were also recognized. Moreover, feline herpesvirus-1 gene was detected by a PCR analysis using a swab obtained from the crusted lesions. Based upon these findings, the present case was considered as having FHV-associated ulcerative dermatitis. Therapy including oral acyclovir and topical recombinant feline interferon omega resulted in marked improvement of the skin and mucosal lesions.

A Study of the Retrovirus-Mediated Transgenic Chicken Production on Chicken Embryos (닭 수정란에서 Retrovirus를 이용한 형질전환 닭 생산 연구)

  • Byun S. J.;Park C.;Kim S. W.;Park J. K.;Chang W. K.;Yang B. S.;Kim T. Y.;Sohn S. H.;Kim S. H.;Jeon I. S.
    • Korean Journal of Poultry Science
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    • v.32 no.4
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    • pp.225-229
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    • 2005
  • Microinjection of recombinant retrovirus beneath the blastoderm of non-incubated chicken embryo is now the most widespread method for generating transgenic chickens, but transgenesis rates are very low. So to improve this problem, we first introduced retrovirus vector carrying RSV-GFP gene to an one-cell embryo culture system. To investigate whether retrovirus could work on an one-cell chicken embryo, we microinjected the concentrated retrovirus stocks into the germinal disc of one cell or stage-X chicken embryos. Analysis of reporter gene expression on day 4 embryos showed that GFP expression was observed in the only stage-X chicken embryo but was not in the one-cell embryo group. These results suggest that retrovirus system is the most efficient method to generate transgenic chickens in the stage-X embryo.

Expression of an artificial gene encoding a repeated tripeptide lysyl-g1utamyl-tryptophan in Tobacco Plant (담배식물체에서 필수아미노산인 lysyl-glutamyl-tryptophan을 암호화하는 인공유전자의 발현)

  • Lee, Soo-Young;Ra, Kyung-Soo;Baik, Hyung-Suk;Park, Hee-Sung;Cho, Hoon-Sik;Lee, Young-Se;Choi, Jang-Won
    • Journal of Life Science
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    • v.12 no.1
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    • pp.96-105
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    • 2002
  • To investigate expression of the artificial gene encoding a repeated tripeptide lysyl-glutamyl-tryptophan in tobacco plant, the plant binary vector, pART404 has been constructed, which contains the duplicated CaMV 35S promoter, an artificial gene coding for repetitive polymer (Lys-Glu-Trp)$_{64}$, and nopaline synthase (nos) terminator. The recombinant expression vector was introduced in Nicotiana tabacum (var. Xanthi) via Agrobacterium tumefaciens-mediated trans-formation. The transgenic calli selected by kanamycin containing medium were then regenerated to whole plants. Southern blot analysis indicated that five transgenic plants (No. 1, 7, 9, 43, 45) showed the hybridizing signals at 1.1 kb of the expected size on EcoRI digestion and each of the transgenic plants contained 1 or 3 copies of the artificial gene inserted into its genome. By northern blot analysis, the size of the hybridized total RNA was estimated to be approximately 1.2 kb and the RNA appeared generally to have the integrity. Western blot indicated that the protein was detected at the position of 33 kDa and the expression level of the polypeptide in the transgenic plant (No. 45) was measured to approximately 0.1% of the total protein.

Growth Suppression by Adenovirus-mediated Gene Transfer of p16/INK4a in Glioma Cell Lines (사람의 신경교종 세포주에서 아데노바이러스 벡터를 이용한 p16/INK4a 유전자 전달에 의한 종양성장 억제)

  • Kim, Mi-Suk;Kwon, Hee-Chung;Kang, Hee-Seog;Park, In-Chul;Rhee, Chang-Hun;Kim, Chang-Min;Lee, Choon-Taek;Hong, Seok-Il;Lee, Seung-Hoon
    • Journal of Korean Neurosurgical Society
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    • v.29 no.4
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    • pp.471-476
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    • 2000
  • Objective : p16/INK4a, a kind of tumor suppressor genes, encodes a specific inhibitor of the cyclin D-dependent kinases CDK4 and CDK6. This prevents the association of CDK4 with cyclin D1, and subsequently inhibits phosphorylation of retinoblastoma tumor suppressor protein(pRb), thus preventing exit from the G1 phase. According to previous reports, over 50% of glioma tissue and 80% of glioma cell lines have been demonstrated inactivation of p16/INK4a gene. The purpose of this study was to determine whether recombinant adenovirus-p16 virus is a suitable candidate for gene replacement therapy in cases of glioma. Methods : Three human glioma cell lines(U251MG, U87MG and U373MG) that express mutant p16 protein were used. Replication-deficient adenovirus was utilized as an expression vector to transfer exogenous p16 cDNA into the cells ; control cells were infected with the Ad-${\beta}$-gal expressing ${\beta}$-galactosidase. To monitor gene transfer and the expression of exogenous genes, we used Western Blotting analysis. Flow cytometry studies of cellular DNA content were performed to determine the cell cycle phenotype of the glioma cells before and after treatment. Results : We showed here that restoration of p16/INK4a expression in p16 negative U87MG, U251MG and partially deleted U373MG by Ad-CMV-p16 induced growth suppression in vitro. Flow cytometric study revealed that Ad-CMV-p16 infected U87MG cells were arrested during the G0-G1 phase of the cell cycle. Expression of p16 transferred by Ad-CMV-p16 in glioma cells was highly efficient and maintained for more than seven days. Conclusions : Our results suggest that Ad-CMV-p16 gene therapy strategy is potentially useful and warrants further clinical investigation for the treatment of gliomas.

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rvH1N1 Neuraminidase Inhibitory Activities of Phenolics from Perilla frutescens (L.) and Their Contents in Cultivars and Germplasm

  • Ha, Tae Joung;Lee, Myoung-Hee;Park, Chang-Hwan;Kim, Jung-In;Oh, Eunyoung;Pae, Suk-Bok;Park, Jae Eun;Kim, Sung-Up;Kwak, Do-Yeon
    • Plant Breeding and Biotechnology
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    • v.6 no.4
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    • pp.404-412
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    • 2018
  • The influenza neuraminidase (NA, E.C. 3.2.1.18), an antiviral, has been the target of high pharmaceutical companies due to its essential role in viral replication cycle. Perilla frutescens (P. frutescens) is used in traditional Chinese medicine for various diseases, such as cold due to wind-cold, headache and cough. In this context, four major polyphenolic compounds including rosmarinic acid-3-O-glucoside (1), rosmarinic acid (2), luteolin (3), and apigenin (4) isolated from P. frutescens were evaluated for their inhibitory effect on recombinant virus H1N1 neuraminidase (rvH1N1 NA). Among the test compounds, rosmarinic acid and luteolin inhibited the rvH1N1 NA with an $IC_{50}$ of 46.7 and $8.4{\mu}M$, respectively. The inhibition kinetics analyzed by the Dixon plots indicated that rosmarinic acid and luteolin were noncompetitive inhibitors and that the inhibition constant, $K_I$, was established as 43.9 and $14.3{\mu}M$, respectively. In addition, 578 genetically diverse accessions and 39 cultivars of P. frutescens were analyzed using HPLC to characterize the diversity of polyphenolic composition and concentration. The individual and total compositions exhibited significant difference (P < 0.05), especially rosmarinic acid which was detected as the predominant metabolite in all accessions (58.8%) and cultivars (62.8%). Yeupsil and Sangback cultivars exhibited the highest rosmarinic acid ($3,393.5{\mu}g/g$) and luteolin ($383.3{\mu}g/g$) content respectively. YCPL177-2 with the high concentration ($889.8{\mu}g/g$) of luteolin may be used as a genetic resource for breeding elite cultivars.

Biological Roles of the Glycan in the Investigation of the Novel Disease Diagnosis and Treatment Methods (신개념 질병 진단 및 치료 연구에 있어서의 당사슬의 생물학적 역할)

  • Kim, Dong-Chan
    • Journal of Life Science
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    • v.28 no.11
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    • pp.1379-1385
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    • 2018
  • Glycans are attached to proteins as in glycoproteins and proteoglycans. They are found on the exterior surface of cells. O- and N-linked glycans are very common in eukaryotic cells but may also be found in prokaryotes. The interaction of cell surface glycans with complementary glycan binding proteins located on neighboring cells, other cell types, pathogens like virus, or bacteria is crucial in biologically and biomedically important processes like pathogen recognition, cell migration, cell-cell adhesion, development, and infection. Their implication in pathological condition, suggests an important role for glycans as disease markers. In addition, a great amount of research has been shown that appropriate glycosylation of a recombinant therapeutic protein is critical for product solubility, stability, pharmacokinetics and pharmacodynamics, bioactivity, and safety. Besides, cancer-associated glycosylation changes often involve sialic acid in glycan branch which play important roles in cell-cell interaction, recognition and immunological response. This review aims at giving a comprehensive overview of the glycan's biological function and describing the relevance among the glycosylation, disease diagnosis and treatment methods. Furthermore, the high-throughput analytic methods available to measure the profile changing patterns of glycan in the blood serum as well as possible underlying biochemical mechanisms.

Comparison of the seroconversion rate after primary hepatitis B vaccination and after revaccination of non-responders in full-term infants according to mother's HBsAg seropositivity (산모의 HBsAg 보유 여부에 따른 만삭아의 B형간염 기본접종 및 무반응자에서 재접종 후 항체 양전율에 대한 연구)

  • Kang, Jang Hee;Moon, Jae Won;Kong, Seung Hyun;Hwang, Kwang Su;Mok, Ji Sun;Lee, Hyeon Jung
    • Clinical and Experimental Pediatrics
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    • v.51 no.11
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    • pp.1165-1171
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    • 2008
  • Purpose : This study aimed to identify the true extent of non-responsiveness in full-term infants born from HBsAg-negative or HBsAg-positive mothers and vaccinated against hepatitis B virus (HBV) at 0, 1, and 6 months of age and to evaluate the effect of revaccination among non-responders. Methods : The study included 716 full-term infants born in 2004-2007. Of 716, 662 infants (A group) were born to HBsAg-negative mothers and 54 infants (B group: 50, except HBsAg-positive infants) were born to HBsAg-positive mothers. All infants were administered DNA recombinant vaccines at 0, 1, and 6 months of age. B group infants received hepatitis B immunoglobulin at birth. Anti-HBs titers were tested at 7-12 and 9-15 months in A and B groups, respectively. Three revaccination doses were administered to non-responders whose anti-HBs titers were under 10 mIU/ml; revaccinated infants were retested at 1-3 months after last vaccination. The association between HBeAg seropositivity of mother and the failure of HBV immunoprophylaxis was evaluated. Results : The seroconversion rates after primary hepatitis B vaccination were higher in A group (94.1%) than in B group (78%, P<0.001). The seroconversion rates were high in revaccinated infants (A group non-responders: 96.9%, B group non-responders: 87.5%). The failure of HBV immunoprophylaxis was significantly associated with maternal HBeAg seropositivity (P<0.001). Conclusion : The seroconversion rates after primary hepatitis B vaccination were low in B group infants. Revaccination of non-responders in B group was very effective. Therefore, anti-HBs testing and revaccination of B group is very important. Revaccination of non-responders in A group was also very effective. Thus, testing the immune status of infants born to HBsAg-negative mothers even after primary hepatitis B vaccination should be considered. However, to realize this, further studies on the cost-effectiveness of anti-HBs testing in healthy full-term infants are necessary.