• KSBB Journal
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• v.21 no.2
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• pp.133-139
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• 2006

In 'solid-phase' PEGylation, the conjugation reaction occurs as the proteins are attached to a solid matrix, and thus it can have distinct advantages over the conventional, solution-phase process. We report a case study: rhIFN-${\alpha}$-2a was first adsorbed to cation exchange resin and then N-terminally PEGylated by aldehyde mPEG of 5, 10, and 20 kD through reductive alkylation. After the PEGylation, salt gradient elution efficiently recovered the mono-PEGylate in a purified form from the unwanted species such as unmodified IFN, unreacted PEG, and others. The mono-PEGylation and its purification were integrated in a single chromatographic step. Depending on the molecular weight of the mPEG aldehyde used, the mono-PEGylation yield ranged 50-64%. We could overcome the major problems of random, or uncontrollable, multi-PEGylation and the post-PEGylation purification difficulties associated with the solution-phase process. N-terminal sequencing and MALDI-TOF MS confirmed that a PEG molecule was conjugated only to the N-terminus. Compared with the unmodified IFN, the mono-PEGylate showed the reduced anti-viral activity as measured by the cell proliferation assay. The bioactivity was reduced more as the higher molecular weight PEG was conjugated. Immunoreactivity, evaluated indirectly by antibody binding activity using a surface plasmon resonance biosensor, also decreased. Nevertheless, trypsin resistance as well as thermal stability was considerably improved.

• Journal of Society of Preventive Korean Medicine
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• v.6 no.2
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• pp.156-167
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• 2002

This experimental study was carried out to evaluate the effects of Kimchi intake of Cordyceps sinensis extract (CDSE) supplementation on cytokine-induction and immune response in mice. To study in experiments using male Balb/c mice fed Kimchi and Kimchi of CDSE supplementation (addition of 2% of total Kimchi weight) containing fed experimental diet during 2 weeks. Experimental mice were fed control diet or diet containing freeze-dried Kimchi at the level of 5%(w/w) or 5% freeze-dried Kimchi with 2% CDSE supplementation. The main ingredient of Kimchi was Korean cabbage and fermentation was carried out at $4^{\circ}C$ for three weeks. Freeze-dried 2% CDSE supplementation was added to Kimchi at the beginning of fennentation. In order to investigate the effect of Kimchi intake of CDSE supplementation (5%Kimchi-2%CDSE), the following was performed; body weight, food intake, hematological parameter, serum level of mouse interleukin-4 (mIL-4) and mouse $interferon-{\gamma}$ $(mIFN-{\gamma})$, and, the percentage of CD3+/CD4+, CD3+/CD8+, B220+ in splenic cells. The results of final body weight, and food diet intake of two Kimchi groups were lower than those of the control group (not supplemented experimental diet). The hematology change obtained from the level of WBC (white blood cell) and platelet were not affected by feeding different dietary regiments, but the level of RBC (red blood cells) HB (hemoglobin), and spleen weight of two Kimchi groups were increased significantly than those of the control group. The serum level of IL-4 and $IFN-{\gamma}$ of two Kimchi groups were increased significantly than those of the control group, also enhanced the percentages of the CD3+/CD4+ and CD3+/CD8+ by 5% freeze-dried Kimchi, and 5%Kimchi-2%CDSE group were 43.9 and 60.1%, and 96.0 and 174% than those of the control group, respectively. From these results, it can be concluded that Kimchi itself has an immuno-stimulatory effect and Kimchi contaning 2% CDSE supplementation has the more pronounced effect in vivo system.

• The Korean Journal of Physiology and Pharmacology
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• v.5 no.1
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• pp.55-63
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• 2001

In macrophages, lipopolysaccharide (LPS) alone or in combination with $interferon-{\gamma}\;(IFN-{\gamma})$ has been shown to release a nitric oxide (NO) through the increase of the transcription of the inducible nitric oxide synthase (iNOS) gene. To investigate the exact intracellular signaling pathway of the regulation of iNOS gene transcription by LPS plus $IFN-{\gamma},$ the effects of protein tyrosine kinase (PTK) inhibitor and protein kinase C (PKC) inhibitors on NO production, iNOS mRNA expression, nuclear $factor-_{\kappa}B\;(NF-_{\kappa}B)$ binding activity and the promoter activity of iNOS gene containing two $NF-_{\kappa}B$ sites have been examined in a mouse macrophage RAW 264.7 cells. LPS or $IFN-{\gamma}$ stimulated NO production, and their effect was enhanced synergistically by mixture of LPS and $IFN-{\gamma}.$ The PTK inhibitor such as tyrphostin reduced LPS plus $IFN-{\gamma}-induced$ NO production, iNOS mRNA expression and $NF-_{\kappa}B$ binding activity. In contrast, PKC inhibitors such as H-7, Ro-318220 and staurosporine did not show any effect on them. In addition, transfection of RAW 264.7 cells with iNOS promoter linked to a CAT reporter gene revealed that tyrphostin inhibited the iNOS promoter activity through the $NF-_{\kappa}B$ binding site, whereas PKC inhibitors did not. Taken together, these suggest that PTK, but not PKC pathway, is involved in the regulation of the iNOS gene transcription through the $NF-_{\kappa}B$ sites of iNOS promoter in RAW 264.7 macrophages by LPS plus $IFN-{\gamma}$.

• Mycobiology
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• v.45 no.4
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• pp.297-311
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• 2017

Saprolegniasis is one of the most devastating oomycete diseases in freshwater fish which is caused by species in the genus Saprolegnia including Saprolegnia parasitica. In this study, we isolated the strain of S. parasitica from diseased rainbow trout in Korea. Morphological and molecular based identification confirmed that isolated oomycete belongs to the member of S. parasitica, supported by its typical features including cotton-like mycelium, zoospores and phylogenetic analysis with internal transcribed spacer region. Pathogenicity of isolated S. parasitica was developed in embryo, juvenile, and adult zebrafish as a disease model. Host-pathogen interaction in adult zebrafish was investigated at transcriptional level. Upon infection with S. parasitica, pathogen/antigen recognition and signaling (TLR2, TLR4b, TLR5b, NOD1, and major histocompatibility complex class I), pro/anti-inflammatory cytokines (interleukin $[IL]-1{\beta}$, tumor necrosis factor ${\alpha}$, IL-6, IL-8, interferon ${\gamma}$, IL-12, and IL-10), matrix metalloproteinase (MMP9 and MMP13), cell surface molecules ($CD8^+$ and $CD4^+$) and antioxidant enzymes (superoxide dismutase, catalase) related genes were differentially modulated at 3- and 12-hr post infection. As an anti-Saprolegnia agent, plant based lawsone was applied to investigate on the susceptibility of S. parasitica showing the minimum inhibitory concentration and percentage inhibition of radial growth as $200{\mu}g/mL$ and 31.8%, respectively. Moreover, natural lawsone changed the membrane permeability of S. parasitica mycelium and caused irreversible damage and disintegration to the cellular membranes of S. parasitica. Transcriptional responses of the genes of S. parasitica mycelium exposed to lawsone were altered, indicating that lawsone could be a potential anti-S. parasitica agent for controlling S. parasitica infection.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.3
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• pp.700-704
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• 2007

In the recent, increased concern has been focused on the pharmacology and clinical utility of herbal extracts and derivatives as a drug or adjunct to chemotherapy and immunotherapy. Here we investigated the role of the extract of Anethi Fructus in the expression of inflammatory mediators, surface molecule, and related receptors in vitro. In murine macrophage RAW 264.7 cells and peritoneal macrophages of C57BL/6N mice, water extract of Anethi Fructus increased the production of secretary tumor necrosis factor (TNF)-a and Nitric oxide (NO), and the expression level of CD14, LPS co-receptor and CD86, co-stimulatory molecule compared to negative natural extract ex vivo. The water extract of Anethi Fructus increased the production of interferon (IFN)-g from splenocytes. Also, water extract of Anethi Fructus increased ConA-induced cell proliferation. These results suggest that water extract of Anethi Fructus may enhance the immune response through immune modulation of macrophage and lymphocytes.

• Molecules and Cells
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• v.37 no.1
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• pp.66-73
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• 2014

Myeloid-derived suppressor cells (MDSCs) play an important role in impairing the function of T cells. We characterized MDSCs in two chronic hepatitis C (CHC) cohorts: a cross-sectional group that included 61 treatment-naive patients with CHC, 14 rapid virologic response (RVR) cases and 22 early virologic response (EVR) cases; and a longitudinal group of 13 cases of RVR and 10 cases of EVR after pegylated-interferon-${\alpha}$/ribavirin treatment for genotype 1b HCV infection. Liver samples from 32 CHC patients and six healthy controls were subjected to immunohistochemical analysis. MDSCs frequency in treatment-naive CHC was significantly higher than in RVR, EVR, or healthy subjects and was positively correlated with HCV RNA. Patients infected with HCV genotype 2a had a significantly higher frequency of MDSCs than those infected with genotype 1b. Decreased T cell receptor (TCR) ${\zeta}$ expression on $CD8^+$ T cells was significantly associated with an increased frequency of MDSCs in treatment-naive CHC patients and was restored by L-arginine treatment in vitro. Increased numbers of liver arginase-$1^+$ cells were closely associated with the histological activity index in CHC. The TCR ${\zeta}$ chain was significantly downregulated on hepatic $CD8^+$ T cells in CHC. During antiviral follow up, MDSCs frequency in peripheral blood mononuclear cells was directly correlated with the HCV RNA load in the plasma and inversely correlated with TCR ${\zeta}$ chain expression in $CD8^+$ T cells in both RVR and EVR cases. Notably, the RVR group had a higher frequency of MDSCs at baseline than the EVR group. Collectively, this study provides evidence that MDSCs might be associated with HCV persistence and downregulation of CD8 ${\zeta}$ chain expression.

• Proceedings of the Korea Multimedia Society Conference
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• 2002.05c
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• pp.501-506
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• 2002

사람은 누구나 잭이나 문서를 읽을 때 중요한 부분에 강조, 해설, 설명을 하기 위해서 표시를 하거나 글을 입력한다. 이와 같이 원본문서에 추가되는 부가 정보를 Annotation이라고 한다[6][7]. Annotation을 이용하면 차후에 원본문서를 재창조하거나 다른 사람이 원본문서를 참조할 경우 과중한 정보의 양을 극복할 수 있으므로[4], 원본문서의 이해도를 향상시킬 수 있다. 따라서, Annotation은 한번 사용하고 그치는 정보가 아닌 재사용할 수 있는 점보임을 의미한다[1,2,3]. 이러한 Annotation 기능을 웹 문서에 적용하게 되면 종이문서에서 얻을 수 있는 장점뿐만 아니라 웹 환경의 특징인 공유[5], 검색[4], 재편집 등의 기능이 가능하다. 이와 관련한 많은 연구가 진행중에 있다. 그러나, 기존의 Annotation 연구는 Anchor 입력된 다수의 Annotation이 무의미한 출력 순서로 제공되고 있으며, 또한 Anchor에 입력된 Annotation의 출력으로 인해 문서 구조가 변경되거나, 가려지는 등의 문제점으로 사용자들이 쉽게 사용 및 이해할 수 있는 Annotation 출력 인터페이스에 대한 연구가 부족한 실정이다. 따라서, 본 논문에서는 Anchor에 입력된 다수의 Annotation들 간의 의미적 순서를 부여하여 보다 적절한 Annotation에 대한 우선 접근이 가능하도록 계층적인 Annotation 우선처리 기법을 제안하고, Annotation 출력으로 인한 문서 변경 문제를 해결하기 위한 유동적인 Annotation 표현 기법을 제안한다. 또한 Annotation이 문서에 부가된 부가정보의 역할을 뿐만 아니라, 다양한 활용이 가능하도록 XML 표준에 기반한 저장 구조를 지원하며, 원본문서와 분리하여 저장한다.속도를 개선시켰고, 국소적인 변형이 있는 패턴과 특징의 수가 다른 패턴의 경우에도 좋은 인식률을 얻었다.r interferon alfa concentrated solution can be established according to the monograph of EP suggesting the revision of Minimum requirements for biological productss of e-procurement, e-placement, e-payment are also investigated.. monocytogenes, E. coli 및 S. enteritidis에 대한 키토산의 최소저해농도는 각각 0.1461 mg/mL, 0.2419 mg/mL, 0.0980 mg/mL 및 0.0490 mg/mL로 측정되었다. 또한 2%(v/v) 초산 자체의 최소저해농도를 측정한 결과, B. cereus, L. mosocytogenes, E. eoli에 대해서는 control과 비교시 유의적인 항균효과는 나타나지 않았다. 반면에 S. enteritidis의 경우는 배양시간 4시간까지는 항균활성을 나타내었지만, 8시간 이후부터는 S. enteritidis의 성장이 control 보다 높아져 배양시간 20시간에서는 control 보다 약 2배 이상 균주의 성장을 촉진시켰다.차에 따른 개별화 학습을 가능하게 할 뿐만 아니라 능동적인 참여를 유도하여 학습효율을 높일 수 있을 것으로 기대된다.향은 패션마케팅의 정의와 적용범위를 축소시킬 수 있는 위험을 내재한 것으로 보여진다. 그런가 하면, 많이 다루어진 주제라 할지라도 개념이나 용어가 통일되지 않고 사용되며 검증되어 통용되는 측정도구의 부재로 인하여 연구결과의 축적이 미비한 상태이다. 따라서, 이에 대한 재고와 새로운 방향

• Journal of Korean Neurosurgical Society
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• v.29 no.4
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• pp.477-484
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• 2000

Objective : Despite advances in the understanding of tumor biology and the tumor immunology, there has been no effective treatment. The Intercellular adhesion molecule-1(ICAM-1) has been shown to be important in interaction involving cells of the immune system and to be upregulated in a number of cell culture systems by cytokines, including immune interferon($IFN-{\gamma}$) and tumor necrosis $factor-{\alpha}$($TNF-{\alpha}$). ICAM-1 has been identified as one of the ligands for lymphocyte function-associated antigen-1(LFA-1). The effectiveness of various cytokines to ICAM-1 induction on cultured human glioblastoma cell lines and potential efficacy of immunotherapy were studied. Method : Human glioblastoma cell lines, U-251 MG, U-373 MG were trypsinized and suspended at $1{\times}10^5cells/ml$ and grown on 8 well chamber slide, the cells were incubated in 0.3ml medium alone or medium containing $IFN-{\gamma}$(1000U/ml) or $TNF-{\alpha}$(250U/ml) or $IFN-{\gamma}$ plus $TNF-{\alpha}$ for 6, 12, 24, 48 and 72 hours. The coverslip were then removed and stained with a 1/30 dilution of anti-ICAM-1 antibody. Result : Surface antigen expression of ICAM-1 was increased by incubating glioblastoma cell lines with $IFN-{\gamma}$ and $TNF-{\alpha}$. Combined effect of $IFN-{\gamma}$ and $TNF-{\alpha}$ has induced more ICAM-1 expression on glioblastoma cell lines. Upregulation of ICAM-1 expression in an established glioblastoma cell line was of greater magnitude and more rapid following incubation with $IFN-{\gamma}$ plus $TNF-{\alpha}$. Surface antigen expression of ICAM-1 was increased for up to 48 hours after cytokine treatment on both cell lines(p<0.05). There was no difference on both cell lines(p>0.05). Conclusion : The results of the present study indicate that ICAM-1 expression in glioblastoma cell lines, U-251 MG and U-373 MG, are induced and enhanced after treatment with $IFN-{\gamma}$ and $TNF-{\alpha}$. Combined effect of $IFN-{\alpha}$ and $TNF-{\gamma}$ is stronger and more rapid than $IFN-{\gamma}$ or $TNF-{\alpha}$ alone.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.3
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• pp.445-451
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• 2016

This study demonstrated the immunological effects of methanol extracts from Doenjang added with wild plants (Pteridium aquilinum and Aster scaber) on bone-marrow derived macrophages and mouse splenocytes. Doenjang (DJ) and wild plant added Doenjang (WPDJ) extracts were treated to bone-marrow derived macrophages (BMDM) and splenocytes, and cell proliferation and cytokine production were measured. Cell proliferation of BMDM and splenocytes was more highly elevated in the WPDJ-treated group compared to the DJ-treated group. Cytokine [tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-6, IL-$1{\beta}$, IL-10, and IL-12] production in BMDM also significantly increased in the WPDJ-treated group. Similarly, in the case of cytokine production in splenocytes, WPDJ treatment highly increased production of Th 1 type cytokines [interferon (IFN)-${\gamma}$ and IL-2] but did not affect production of Th 2 type cytokines (IL-4). These results suggest that wild plants could improve the immunomodulatory activity of Doenjang and may be effective for the development Doenjang.

• Journal of Korean Medical Science
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• v.33 no.47
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• pp.292.1-292.10
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• 2018

Background: We investigated the incidence of active tuberculosis among patients with inflammatory bowel disease (IBD) treated with tumor necrosis factor (TNF) inhibitors, with or without latent tuberculosis infection (LTBI). Methods: The study was performed at a Korean tertiary referral center between January 2011 and June 2017. In total, 740 patients with IBD who underwent LTBI screening tests and were followed-up for ${\geq}1$ year after TNF inhibitor treatment initiation were enrolled. LTBI was detected on the basis of tuberculin skin test results, interferon-gamma release assay results, chest X-ray findings, and previous tuberculosis treatment history. The patients were classified into LTBI (n = 84) or non-LTBI (n = 656) group. The risk of developing tuberculosis in each group was assessed on the basis of standardized incidence ratio (SIR) and 95% confidence interval (CI) for active tuberculosis. Results: Mean patient age was 33.1 years, and patients with Crohn's disease were predominant (80.7%). Within 1 year after the initiation of TNF inhibitor treatment, 1 patient in the LTBI group (1/84; 1.2%) and 7 patients in the non-LTBI group (7/656; 1.1%) developed active tuberculosis. The overall 1-year incidence of tuberculosis among the patients was significantly higher than that among the general population (SIR, 14.0; 95% CI, 7.0-28.0), and SIR was not affected by LTBI status (LTBI group: 14.5, 95% CI, 2.0-102.6; non-LTBI group: 14.0, 95% CI, 6.7-29.4). Conclusion: Patients with IBD undergoing TNF inhibitor treatment showed a higher 1-year incidence of tuberculosis than the general population irrespective of LTBI status.