• BMB Reports
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• 제28권4호
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• pp.353-358
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• 1995

Epitope tagging is the process of fusing a set of amino acid residues that are recognized as an antigenic determinant to a protein of interest. Tagging a protein with an epitope facilitates various immunochemical analyses of the tagged protein with a specific monoclonal antibody. The monoclonal antibody H8 has subtype specificity for an epitope derived from the preS2 region of hepatitis B virus surface antigen. Previous studies on serial deletions of the preS2 region indicated that the preS2 epitope was located in amino acid residues 130~142. To test whether the amino acid sequence in this interval is sufficient to confer on proteins the antigenicity recognizable by the antibody H8, the set of amino acid residues in the interval was tagged to the amino terminal of ${\beta}$-galactosidase and to the carboxyl terminal of the truncated $p56^{lck}$ fragment. The tagged ${\beta}$-galactosidase, expressed in Escherichia coli, maintained the enzymatic activity and was immunoprecipitated efficiently with H8. The tagged $p56^{lck}$ fragment, synthesized in an in vitro translation system, was also immunoprecipitated specifically with H8. These results demonstrate that the amino acid sequence of the preS2 region can be used efficiently for the epitope tagging approach.

• Journal of Ginseng Research
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• 제19권2호
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• pp.117-121
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• 1995

We studied the effects of ginsenoside-$Rg_1$ (G-$Rg_1$) extracted from Panax ginseng C.A. Meyer on $p56^{kk}$ kinase and cell proliferation in Jurkat T cells. $p56^{kk}$ was maximally activated within 5 min after the treatment of 16.7 $\mu\textrm{g}$/ml of G-$Rg_1$ increasing the activity by 1.2-2 times relative to untreated control, thereafter its activity was gradually decreased to the level of untreated control. The action of EGTA on the kinase was altered by the addition of G-$Rg_1$, accompanying the band shift of $p56^{kk}$ to $p60^{kk}$. In addition, G-$Rg_1$promoted cell proliferation in a concentration-dependent manner. These results suggest that G-$Rg_1$ may be involved in T cell receptor-CD3 (TCR) signaling via the activation of $p56^{kk}$ and the chance of cellular calcium concentration.

• BMB Reports
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• 제31권4호
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• pp.355-363
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• 1998

MHC unrestricted, antigen nonspecific killing by $CD4^+$ T-cells against virally-infected target cells was induced following cross-linking of CD4 molecules. The cytotoxicity of antibody-activated $CD4^+$ T-cells was abolished by genistein (4',5,7-trihydroxyisoflavone), a protein tyrosine kinase (PTK) inhibitor, but not by H-7, a protein kinase C (PKC) inhibitor. Genisteintreated human or bovine peripheral blood $CD4^+$ T-cells lacked PTK activity and failed to kill virally-infected target cells even after cross-linking of CD4 molecules. The cross-linking of CD4 molecules did not induce effector cell proliferation or the transcription of TNF ${\beta}$. TNF ${\beta}$ synthesis was up-regulated by incubating antibody activated effector cells with bovine herpesvirus type 1 (BHV-1) infected D17 target cells. Anti-TNF ${\beta}$ antibody partially abrogated direct effector cell-mediated antiviral cytotoxicity. On the other hand, this antibody effectively neutralized antiviral activity of effector and target cell culture supernatants against BHV-1 infected D17 cells. The inhibition level of the antiviral activity by the antibody was dependent on effector and target cell ratio. These findings have importance to define the mechanisms of how CD4 cytotoxic cells control viral infection.

• 한국동물학회:학술대회논문집
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• 한국동물학회 1996년도 한국생물과학협회 국제학술대회
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• pp.260.2-260
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• 1996

No Abstract, See Full Text

• Animal cells and systems
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• 제3권3호
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• pp.331-336
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• 1999

p56$^{Ick}$, a cytoplasmic protein tyrosine kinase of the src family, is non-covalently associated with the cell surface coreceptors CD4 and CD8, which are expressed on thymocytes and mature T cells. The coreceptor protein plays an important role during the differentiation of thymocytes and the activation of T cells. DNA constructs were designed to study the roles of CD4 and CD8 during the differentiation of thymocytes. One is a chimeric cDNA which consists of coding regions for the extracellular domain of CD8a and the transmembrane and cytoplasmic domain of CD4. The other is the same chimeric cDNA but with a point mutation converting Cys to Ala in the Ick-binding site to disrupt the association. We confirmed that the CD8a/CD4 chimeric molecule bound to Ick more efficiently than the wild type CD8a protein. However, the chimeric protein with the Cys$leftrightarro$Ala mutation did not associate with Ick. The results suggest a possibility that the CD8a/CD4 chimeric protein may behave like a CD4 protein in associating with Ick and that it may deliver a signal inside the cell in a similar manner, Analysing effects of the mutant CD8a/CD4 chimeric protein expression in developing thymocytes will elucidate the role of Ick during the determination of CD4/CD8 cell lineages.

• 대한간호학회지
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• 제15권1호
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• pp.30-43
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• 1985

Liver cirrhosis is the common cause of death in Korea today. But, if liver cirrhosis Patients were treated in the early stage of the disease Process, they can have a chance to carry their daily lives with prescribed medical and nursing regimens. Each patient has different health beliefs that there is a different Sick Role Behavior in the process of treatment. In order to increase and control the desired patient's Sick Role Behavior, it is important for nurses to understand the health beliefs influencing Sick Role Behavior. The purpose of this study was to determine factors influencing Sick Role Behavior and provide objective and scientific data to health education, treatment and nursing care. The subjects for this study were 80 Liver Cirrhosis patients selected from in and out patients of the medical department of four University Hospitals in Seoul, Won Joo and Mok Po city. Data was collected from Sep. 18, to Oct. 15, 1984. The measurement tool was the questionaire that developed by the investigator from the literature review based on Health Belief Model. The data Collection was done by interview. Analysis of data was done by use Mean, S.D., ANOVA, Pearson Correlation Coefficient and Stepwise Multiple Regression. The result of study were as follows: 1. The significant influencing variables on the Liver Cirrhosis Patient's Sick Role Behavior in general characteristics were Sex, Marital Status, Educational levels, Family's income and Duration after diagnosis. 2. Between the Sick Role Behavior and Health Belief Model, a) The first hypothesis that the stronger degree of Health Motivation, the higher degree of Liver Cirrhosis Patient's Sick Hole Behavior was supported (r=0.7892, p=0.0000). b) The second hypothesis that the higher degree of perceived susceptibility, the higher degree of Liver Cirrhosis Patients' Sick Role Behavior was supported (r=0.6383, p=0.0000) c) The third hypothesis that the higher degree of perceived severity, the higher degree of Liver Cirrhosis Patients' Sick Role Behavior was supported (r=0.5869, p=0.0000). d) The fourth hypothesis that the higher degree of perceived benefit, the higher degree of Liver Cirrhosis patient's Sick Role Behavior was supported (r=0.7535, p=0.0000). e) The fifth hypothesis that the lower degree of perceived barrier, the higher degree of Liver Cirrhosis Patient's Sick Role Behavior was supported(r=-9.7709, p=0.0000) f) The sixth hypothesis that the higher degree of knowledge in Disease, the higher degree of Liver Cirrhosis patients'lck Role Behavior was supported (r=0.7538, p=0.0000), g) In the correlation among variables, it was found positive correlation except that perceived barrier was negatively correlated. In the Stepwise Multiple Regression and Independent Variables, the factor“Health Motivation”could account for Sick Role Behavior in 62.28% of the Sample (F=128. 786, p<0.01). When the factor“perceived barrier”is added to this, it account for 70.38% of Sick Role Behavior (F=93.479, p <0.01) and the factor“knowledge in disease”is also included, it account for 74.78% of Sick Role Behavior (F=75.131, p <0.01). Finally, when the factor“perceived susceptibility”is included, it account for 75.03% of Sick Role Behavior (F=56.329, p <0.01).

• 대한의생명과학회지
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• 제2권2호
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• pp.175-185
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• 1996

모든 진핵세포에 존재하며 세포의 성장 및 분화에 주로 관계되는 신호전달물질의 하나인 Mitogen-activated protein(MAP)kinase의 mitogen에 의한 핵내 활성화와 기질 인산화에 대해 알아보기 위해 본 실험을 수행하였다. P388세포를 10% fetal bovine serum이 첨가된 DMEM배지에 배양한 후, 혈청이 들어있지 않은 배지에서 24시간 더 배양하고 serum 및 PMA를 농도별로 처리하여 세포성 장을 위한 최적 농도를 확인한 결과 serum은 5-20% 농도에서 세포성장을 촉진시켰고 PMA는 실험한 모든 농도에서 세포성장을 거의 촉진시키지 못하는 경향을 확인하였다. 이어 P388 세포를 serum 및 PMA로 10 분간 활성화하여 파쇄한후 세포질분획과 핵분획으로 분리하여 각 분획을 10% gel 상에서 전기영동 하여 nitrocellulose paper에 옳긴 후 anti-ERKI antibody를 이용해 확인해본 결과 serum, PMA로 처리된 세포 모두에서 MAP kinase의 핵내 이동이 관찰되었으며 특히 세포질 내에 주로 존재하는 42, 44 Kd의 MAP kinase isoform중 42 Kd의 isoform이 주로 핵내로 이동되는 것이 관찰되었다. MAP kinase의 기질인산화 실험을 위해 serum으로 활성화시킨 세포를 파쇄하여 SP-sephadex C-50, Phenyl superose, Mono Q column의 순서로 chromatography를 시 행하여 MAP kinase를 부분분리 하였다. 이와 같이 얻은 MAP kinase를 가지고 면역 T세포에 존재하는 tyrosine kinase인 $p56^{lck}$ 의 N-terminal peptide로 구성된 GST-fusion protein에 대한 인산화를 확인하였다. 또한 세포에서 분리한 MAP kinase를 가지고 transcription factor의 하나인 c-Jun protein에 대한 인산화실험을 실시한 결과 MAP kinase에 의해 인산화 됨이 확인되었다. 이상의 결과를 통해 P388세포는 (1)세포 성장시 외부 신호를 G-protein-coupled receptor/protein kinase C/MAP kinase의 경로보다는 주로 tyrosine kinase receptor protein/Ras/MAP kinase의 경로를 이용하여 핵으로 전달하는 것으로 추측되 며 (2) mitogen의 처리로 활성화된 MAP kinase중 주로 42 Kd isoform이 핵내로 이동하고, 분리한 MAP kinase가 GST-fusion protein과 transcription factor인 c-Jun을 모두 인산화 시키는 결과로 보아 MAP kinase의 isoform에 따라 표적 compartment가 다르고 결과적으로 표적 기질에 차이가 있을지 모른다고 간접적으로 추론할 수 있다.

• Clinics in Shoulder and Elbow
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• 제5권2호
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• pp.88-97
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• 2002

The purpose of this study was to compare the outcomes between arthroscopir repair and mini-open repair of medium and large rotator cuff tears in which arthroscopic repair was technically unsuccessful. We evaluated 76 patients of full-thickness rotator cuff tears, among them 42 patients had all-arthroscopic and 34 patients had mini-open salvage repairs. Patients who had acromioclavicular arthritis, subscapularis tear, or instability were excluded. There were 39 males and 37 females with mean age of 56 years (range,42 to 75 years). At a mean follow-up of 39 months (range, 24 to 64 months), the results of both groups were compared with regard to the UCLA and ASES shoulder rating scale s. Shoulder scores improved in all ratings in both groups (p > 0.05). Overall, sixty-six patients showed excellent or gr)of and ten patients showed fair or poor scores by the UCLA scale. Seventy-two patients satisfactorily returned to prior activity. Four showed unsatisfactory return. The range of motion, strength, and patient's satisfaction were improved postoperatively. There were no difference in shoulder scores, pain, and activity return between the arthroscopic and mini-open salvage groups (p > 0.05). However, Patients with larger size tear showed lower shoulder scores and less predictive recovery of the strength and function (p < 0.05). Postoperative pain was not different with respect to the size of the tear (p : 0.251). Arthroscopic repair of medium and large full-thickness rotator cuff tears had iln equal outcome to technically unsuccessful arthroscopic repairs, which were salvaged by conversion to a mini- open repair technique. Surgical outcome depended on the size of the tear, rather than the method of repair.