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Definition of the peptide mimotope of cellular receptor for hepatitis C virus E2 protein using random peptide library (Random peptide library를 이용한 C형 간염바이러스 E2 단백질 세포막 수용체의 peptide mimotope 규명)

  • Lee, In-Hee;Paik, Jae-Eun;Seol, Sang-Yong;Seog, Dae-Hyun;Park, Sae-Gwang;Choi, In-Hak
    • IMMUNE NETWORK
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    • v.1 no.1
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    • pp.77-86
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    • 2001
  • Background: Hepatitis C virus(HCV), a family of Flaviviridae, has a host cell-derived envelope containing a positive-stranded RNA genome, and has been known as the maj or etiological agent for chronic hepatitis, hepatic cirrhosis, and hepatocellular carcinoma. There remains a need to dissect a molecular mechanism of pathogenesis for the development of therapeutic and effective preventive measure for HCV. Identification of cellular receptor is of central importance not only to understand the viral pathogenesis, but also to exploit strategies for prevention of HCV. This study was aimed at identifying peptide mimotopes inhibiting the binding of E2 protein of HCV to MOLT-4 cell. Methods: In this study, phage peptide library displaying a random peptides consisting of 7 or 12 random peptides was employed in order to pan against E2 protein. Free HCV particles were separated from the immune complex forms by immunoprecipitation using anti-human IgG antibody, and used for HCV-capture ELISA. To identify the peptides inhibiting E2-binding to MOLT-4 cells, E2 protein was subj ect to bind to MOLT-4 cells under the competition with phage peptides. Results: Several phage peptides were selected for their specific binding to E2 protein, which showed the conserved sequence of SHFWRAP from 3 different peptide sequences. They were also able to recognize the HCV particles in the sera of HCV patients captured by monoclonal antibody against E2 protein. Two of them, showing peptide sequence of HLGPWMSHWFQR and WAPPLERSSLFY respectively, were revealed to inhibit the binding of E2 protein to MOLT-4 cell efficiently in dose dependent mode. However, few membrane-associated receptor candidates were seen using Fasta3 programe for homology search with these peptides. Conclusion: Phage peptides containing HLGPWMSHWFQR and WAPPLERSSLFY respectively, showed the inhibition of E2-binding to MOLT-4 cells. However, they did not reveal any homologues to cellular receptors from GenBank database. In further study, cellular receptor could be identified through the screening of cDNA library from MOLT-4 or hepatocytes using antibodies against these peptide mimotopes.

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Inhibitory Effects of Chicken Egg Yolk Antibody on Infection of Escherichia coli in Macrophage

  • Lee, Jin-Ju;Kim, Dong-Hyeok;Lim, Jeong-Ju;Kim, Dae-Geun;Kim, Gon-Sup;Min, Won-Gi;Lee, Hu-Jang;Rhee, Man-Hee;Chang, Hong-Hee;Kim, Suk
    • Journal of agriculture & life science
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    • v.46 no.2
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    • pp.107-114
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    • 2012
  • The present study evaluated the potential use of immunoglobulin prepared from egg yolk of chickens immunized with Escherichia coli K88 (IgY-Ec) in the control of E. coli K88 infection in RAW 264.7 murine macrophage. The binding activity of IgY-Ec against E. coli K88 surface protein was more specific and increased than control IgY. In infection assay of E. coli in macrophage, the specific IgY-Ec to E. coli K88 remarkably inhibited the phagocytic activity comparing to nonspecific IgY (p<0.001). In adherence assay, bacterial adhesion on macrophage cells was definitely reduced by preincubation of IgY-Ec compared with nonspecific IgY (p<0.05). These findings suggested that IgY-Ec have the protective effects against pathogens and IgY-based diets may have potential benefits for preventing or treating various infections in domestic animals.

Effect of Tongqiao-tang on OVA Induced Allergic Rhinitis Mouse Model

  • Lee, Kyu-Jin;Nam, Hae-Jeong;Kim, Yoon-Bum
    • The Journal of Korean Medicine
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    • v.29 no.5
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    • pp.96-103
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    • 2008
  • Objectives : Tongqiao-tang(TQT) has been commonly used for the treatment of common cold, rhinitis etc. Nowadays, TQT becomes one of the most frequently used medicines for allergic rhinitis, but the mechanism of TQT in vivo isn't investigated yet. This study was performed to investigate the effect of TQT on OVA-induced allergic rhinitis mouse model by calculating serum cytokines and IgE. Methods : 8 weeks aged male BALB/c mice were divided into three groups: the normal group, the control group and the medicated group (the TQT group). Each group was consisted of 15 mice. The TQT group was administered TQT extract orally one time a day (1g/kg) from the $1^{st}$ day of experiment till the $26^{th}$ day. The control group and the normal group were administered normal saline by the same method of the TQT group. To induce the allergic rhinitis in the control group and the TQT group, mice of each group were sensitized intraperitoneally with ovalbumin (OVA) solution at the $1^{st}$, the $7^{th}$ and the $14^{th}$ day. After then, intranasal sensitization was performed by dropping 0.1% OVA solution in nasal cavity at the $22^{th}$, the $24^{th}$ and the $26^{th}$ day. At the $27^{th}$ day, the mice were killed and the changes of interferon-${\gamma}$, interleukin-4, interleukin-5, total IgE and OVA-specific IgE were checked. Results : IFN-${\gamma}$ was increased 36% more in the TQT group than that in the control group. IL-4, IL-5, the total IgE and OVA-specific IgE were decreased in the TQT group as compared with the control group and these results were statistically significant. Conclusions : Considering the above experimental results, this study showed that TQT could reduce the allergic reaction in allergic rhinitis. Advanced studies are required to investigate the further mechanisms of TQT.

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Development of multiplex polymerase chain reaction for the detection of vancomycin resistant genotypes and Enterococcus Sp.-specific genes (장구균의 vancomycin 내성 유전자와 종 특이유전자의 검출을 위한 Multiplex polymerase chain reaction 개발)

  • Cho, Yun-Sang;Lee, Hee-Soo;Kim, Jong-Man;Ahn, Jong-Sam;Ryu, Pan-Dong;Park, Yong-Ho;Yoo, Han-Sang;Lee, Mun-Han
    • Korean Journal of Veterinary Research
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    • v.43 no.1
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    • pp.103-112
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    • 2003
  • A multiplex PCR assay, which allows simultaneous detection of vancomycin resistant genotypes and Enterococcus species-specific genes, was developed. Vancomycin resistant enterococci (VRE) from chickens and humans could be detected for vanA, vanB, vanC-1, vanC-2, $ddl_{E.faecium}$ and $ddl_{E.faecalis}$ by multiplex PCR. Eight isolates of VRE from humans (n=11) had $ddl_{E.faecium}$ and vanA, and 3 isolates of the VRE had $ddl_{E.faecium}$ and vanB. One isolate of VRE from chickens (n=6) had $ddl_{E.faecium}$ and vanA, and 5 isolates of the VRE had only vanA. E. faecium, E. faecalis, E. gallinarum and E. casseliflavus were also confirmed for the species-specific gene by multiplex PCR. This multiplex PCR could detect E. faecium, E. faecalis, E. gallinarum, E. casseliflavus, vanA, vanB, vanC-1 and vanC-2, simultaneously. The PCR assay established in the present study can be an alternative to time-consuming biochemical tests and antibiotic susceptibility tests of Enterococcus spp.

Neurogenesis and neuronal migration of dopaminergic neurons during mesencephalon development in mice

  • Kim, Mun-ki;Lee, Si-Joon;Vasudevan, Anju;Won, Chungkil
    • Journal of Biomedical and Translational Research
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    • v.19 no.4
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    • pp.125-129
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    • 2018
  • Dopaminergic neurons are one of the major neuronal components in the brain. Mesencephalon dopamine (DA) neurogenesis takes place in the ventricular zone of the floor plate, when DA progenitors divide to generate postmitotic cells. These cells migrate through the intermediate zone while they differentiate and become DA neurons on reaching the mantle zone. However, neurogenesis and neuronal migration on dopaminergic neurons remain largely unexplored in the mesencephalon development. This study presents neurogenesis and neuronal migration patterns of dopaminergic neurons during mesencephalic development of the mouse. Neurons from embryonic day (E) 10-14 were labelled by a single injection of 5-bromodeoxyuridine and immunohistochemistry was performed. The neurogenesis occurred mainly at the E10 and E11, which was uniformly distributed in the mesencephalic region, but neurons after E13 were observed only in the dorsal mesencephalon. At the postnatal day 0 (P0), E10 generated neurons were spread out uniformly in the whole mesencephalon whereas E11-originated neurons were clearly depleted in the red nucleus region. DA neurons mainly originated in the ventromedial mesencephalon at the early embryonic stage especially E10 to E11. DA neurons after E12 were only observed in the ventral mesencephalon. At E17, E10 labelled neurons were only observed in the substantia nigra (SN) region. Our study demonstrated that major neurogenesis occurred at E10 and E11. However, neuronal migration continued until neonatal period during mesencephalic development.

Qi-deficiency in the Patients with Allergic Disorders

  • Yei, Young Chul;Jin, Chul;Choi, Won Woo;Hwang, Jae Woong;Jang, Hyeung Jin;Kim, Bum Joon;Lim, Kook Jin;Kwon, Seungwon;Cho, Seung Yeon;Park, Seong Uk;Park, Jung Mi;Ko, Chang Nam;Moon, Sang Kwan;Cho, Ki Ho;Jung, Woo Sang
    • The Journal of the Society of Stroke on Korean Medicine
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    • v.16 no.1
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    • pp.19-24
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    • 2015
  • ■ Introduction Allergy refers a hypersensitive immune response to the various environmental factors and Qi is considered as vit al energy to defend oneself from environments in Traditional Chinese Medicine. Therefore, this study was performed to assess the relationship between allergy and Qi-deficiency ■ Methods We enrolled the subjects with allergic disorders and the healthy controls without allergic disorders. We got the information on the general characteristics, and the score of Qi-deficiency from all of the subjects, and measured their total Immunoglobulin E(IgE) level. ■ Results There were one hundred forty three subjects with allergic disorders and 211 of the control. The educational level, total IgE, and the scores of the Qi-deficiency were higher in the allergy group than in the control. ■ Conclusion Allergic subjects tend to have more severe Qi-deficiency. This could suggest the clinical effectiveness of some herbal medicines for Qi-deficiency on allergic disorders.

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Transduction of eGFP Gene to Human Embryonic Stem Cells and Their Characterization (인간 배아줄기세포로의 eGFP 유전자 도입 및 특성 분석)

  • Kim, Yoon-Young;Ku, Seung-Yup;Park, Yong-Bin;Oh, Sun-Kyung;Moon, Shin-Yong;Choi, Young-Min
    • Clinical and Experimental Reproductive Medicine
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    • v.36 no.4
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    • pp.283-292
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    • 2009
  • Objective: Human embryonic stem cells (hESCs) can proliferate indefinitely and differentiate into all kinds of cell types in vitro. Therefore, hESCs can be used as a cell source for cell-based therapy. Transduction of foreign genes to hESCs could be useful for tracing differentiation processes of hESCs and elucidation of gene function. Thus, we tried to introduce enhanced green fluorescent protein (eGFP) gene to hESCs, XX and XY cell lines in this study. Methods: Lentivirus containing eGFP was packaged in 293T cells and applied to hESCs to transduce eGFP. Expression of transduced eGFP was evaluated under the fluorescence microscope and eGFP positive population was analyzed by FACS. Expression of undifferentiation state markers such as Oct4, Nanog, SSEA4 and Tra-1-81 was examined by RT-PCR and/or immunofluorescence in eGFP-hESCs after transduction. In addition, the ability of eGFP-hESCs to form embryoid bodies (EBs) was tested. Results: eGFP was successfully transduced to hESCs by lentivirus. eGFP expression was stably maintained up to more than 40 passages. eGFP-hESCs retained expression patterns of undifferentiation state markers after transduction. Interestingly, disappearance of transduced eGFP was notably observed during spontaneous differentiation of eGFP-hESCs. Conclusion: We established eGFP expressing hESC lines using lentivirus and showed the maintenance of undifferentiation characteristics of these eGFP-hESCs. This reporter-containing hESCs could be useful for tracing the processes of differentiation of hESCs and other studies.

Differential expression of the metastasis suppressor KAI1 in decidual cells and trophoblast giant cells at the feto-maternal interface

  • Koo, Tae Bon;Han, Min-Su;Tadashi, Yamashita;Seong, Won Joon;Choi, Je-Yong
    • BMB Reports
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    • v.46 no.10
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    • pp.507-512
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    • 2013
  • Invasion of trophoblasts into maternal uterine tissue is essential for establishing mature feto-maternal circulation. The trophoblast invasion associated with placentation is similar to tumor invasion. In this study, we investigated the role of KAI1, an anti-metastasis factor, at the maternal-fetal interface during placentation. Mouse embryos were obtained from gestational days 5.5 (E5.5) to E13.5. Immunohistochemical analysis revealed that KAI1 was expressed on decidual cells around the track made when a fertilized ovum invaded the endometrium, at days E5.5 and E7.5, and on trophoblast giant cells, along the central maternal artery of the placenta at E9.5. KAI1 in trophoblast giant cells was increased at E11.5, and then decreased at E13.5. Furthermore, KAI1 was upregulated during the forskolin-mediated trophoblastic differentiation of BeWo cells. Collectively, these results indicate that KAI1 is differentially expressed in decidual cells and trophoblasts at the maternal-fetal interface, suggesting that KAI1 prevents trophoblast invasion during placentation.