• Title/Summary/Keyword: O-antigen

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Preparation of α-Linked 6-Deoxy-D-altro-heptopyranosidic Residues

  • 신영숙;천근호;Shin, E. Nam;Gerald O. Aspinall
    • Bulletin of the Korean Chemical Society
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    • v.16 no.7
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    • pp.625-630
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    • 1995
  • α-Linked D-altropyranosidic derivatives were obtained by configurational change at C-3 of α-D-mannopyranosides as the key step in preparation of allyl and methyl α-D-glycopyranosides of 6-deoxy-D-altro-heptose. The manno-altro conversion was effected by sequential reactions of Swern oxidation and stereoselective borohydride reduction. Allyl 4,6-O-benzylidene-2-O-p-methoxybenzyl-α-D-mannopyranoside was transformed to the corresponding altropyranoside via 3-oxo-arabino-hexopyranoside. Allyl 7-O-benzyl-6-deoxy-3,4-O-isopropylidene-α-D-altro-heptopyranoside has been prepared as a glycosyl acceptor to be coupled with β-D-GlcpNAc-(1→3)-α-D-Galp glycosyl donor for the synthesis of an O-antigen repeating unit of Campylobacter jejuni serotypes O:23 and O:36. Stereoselective borohydride reduction also succeeded in yielding methyl 2,4,7-tri-O-benzyl-6-deoxy-α-D-altro-heptopyranoside from the corresponding 3-oxo-α-D-arabino-heptopyranoside. C-6 Homologation was achieved by sequential reactions of cyanide displacement of 6-sulphonates, reduction of the resulting heptopyranosidurononitrile with diisobutylaluminum hydride, hydrolysis of the imine, and further reduction with sodium borohydride.

Endogenous Proteinaceous Inhibitor for Protein Methylation Reactions

  • Paik, Woon-Ki;Lee, Hyang-Woo;Kim, Sangduk
    • Archives of Pharmacal Research
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    • v.10 no.3
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    • pp.193-196
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    • 1987
  • Protein methylation occurs ubiquitously in nature and involves N-methylation of lysine, arginine, histidine, alanine, proline and glutamine, O-methylesterfication o dicarboxylic acids, and S-methylation of cysteine and methionine. In nature, methylated amino acids accur in highly specialized proteins such as histones, flagella proteins, myosin, actin, ribosomal proteins. hn RNA-bound protein, HMG-1 and HMG-2 protein, opsin, EF-Tu, EF-$1\alpha$, porcine heart citrate synthase, calmodulin, ferredoxin, $1\alpha$-amylase, heat shock protein, scleroderma antigen, nucleolar protein C23 and IF-3l.

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Studies on the Applicability of Various Antigen Preparations in the Immunoblot Diagnosis of Cysticercosis (Immunoblot법을 이용한 낭미충증(囊尾蟲症)진단에 있어서 각종 항원(抗原)의 적용가능성(適用可能性) 검토(檢討)에 관한 연구(硏究))

  • Koh, Young-Tae;Joo, Kyoung-Hwan;Chung, Myung-Sook;Rim, Han-Jong
    • Journal of agricultural medicine and community health
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    • v.16 no.1
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    • pp.79-89
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    • 1991
  • A systematic study was conducted to identify and isolate a serologically pertinent antigen with high specific activity and low cross reactivity from Cysticercus parenchymal antigen. Differential centrifugation of the homogenate yield three particulate and one soluble fractions ; the $480{\times}G$ pellets($CyL_2$), the $7650{\times}G$ pellet($CyL_3$), the $100000{\times}G$ pellet($CyL_4$), and $100000{\times}G$ supernatant($CyL_6$). We compared antigenicity of these antigens to that or cystic fluid antigens($CyF_1$), saline extract of cystic wall($CyL_1$), and n-butanol treated $GyL_4$ antigen ($CyL_6$) based on SDS-PAGE and immunoblot techniques. The data obtained were as follows : 1) The ratio of O.D. value of ELISA against cysticercosis positive pool sera to that of negative pool sera was highest when using $CyF_1$ as antigen. However the ratio was relatively low in case of $CyL_{3.4}$ and $CyL_5$. 2) We have noted in previous paper that most strong antigenic activities are present in 63Kd band with low cross reactivities. An effective serologic reagent must contain components that are recognized by most infected sera. 63Kd band met this criteria and could be considered as a reliable band for the diagnosis of cysticercosis. As far as 63Kd band concern, $CyL_5$ showed most strong activities without disturbance of cross reaction by EITB in spite of low applicability to microplate ELISA. 3) $CyL_5$ could detect the serum antibody of cysticercosis even in very low titers, around cut-off values of microplate ELISA, by immunoblot. It also could detect the cross reactivities of Echinococcus species, which showed high absorbance value in micro plate ELISA and some sparganosis cases. Further purification of this antigen will be able to represents a antigen that can be used in the diagnosis of cysticercosis.

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Acute Epstein-Barr Virus Hepatitis in a 32 Month Old Female Manifesting as Henoch-Sch$\ddot{o}$nlein Purpura

  • Kim, Hee Jin;Jung, Su Jin;Lee, Jun Ho
    • Childhood Kidney Diseases
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    • v.19 no.1
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    • pp.39-42
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    • 2015
  • Henoch-Sch$\ddot{o}$nlein purpura can result from exposure to an antigen after infection with several types of organisms. However, Henoch-Sch$\ddot{o}$nlein purpura caused by a primary Epstein-Barr virus infection has been rarely reported. Here, we report the case of a 32-month-old female patient who presented with Henoch-Sch$\ddot{o}$nlein purpura. Based on abnormal liver function test results and positive results for Epstein-Barr virus infection markers, a diagnosis of Epstein-Barr virus hepatitis manifesting as Henoch-Sch$\ddot{o}$nlein purpura was made. Treatment with methyl-prednisolone and hydration improved the symptoms, and a switch to oral steroids was effective in completely alleviating the purpura. No recurrence was noted and no liver function abnormalities were detected during the follow up period.

Gastrointestinal Carriage of Escherichia coli with Hemagglutination Activity and Colonization Factor Antigen I and its Relation to Diarrhea (Mannose-resistant Hemagglutination(MRHA) 및 Colonization Factor Antigen I(CFA I)을 표현하는 대장균의 장관내 존재와 설사증 발현과의 관계)

  • Ro, Sung-Hyun;Kim, Kyung-Hee;Cho, Yaug-Ja;Suh, Inn-Soo
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.2
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    • pp.147-153
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    • 1987
  • Colonization factor antigen I(CFA I) has been shown to be one of several virulence factors that promote attachment of enterotoxigenic E. coli(ETEC) to small intestinal epithelial cells of humans. The ability of ETEC to produce mannose-resistant hemagglutination(MRHA) of human blood group A has been used to detect CFA I. To determine gastrointestinal carriage in Korean children of E. coli with MRHA and CFA I, 116 strains of E. coli from diarrheal children admitted to Hanyang University Hospital were examined for MRHA of human erythrocytes and the presence of CFA I. Of 45 ETEC strains, 18(40%) gave a positive MRHA($MRHA^+$) and eight(18%) were positive for CFA I(CFA $I^+$). ETEC with CFA I were all heat-stable enterotoxin(ST) producers and two of these strains were of serogroups $O_{25}$. Of 17 classic enteropathogenic E. coli(EPEC), 7(41%) were $MRHA^+$ but all were negative for CFA I(CFA $I^-$). Of 30 enteroadherent E. coli(EAEC) strains, 11(37%) were $MRHA^+$ and one was CFA $I^+$. Of 24 nonpathogenic E. coli, 4(17%) were $MRHA^+$ but all were CFA $I^-$. It was shown that MRHA was common in all strains of E. coli, CFA I was limited only to ST producing ETEC and EAEC; although MRHA is a useful screening procedure, serologic tests seem to be necessary to comfirm CFA I production. CFA I was associated with a lower proportion of ETEC isolates in Korea than has been reported for other locations.

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High $T_c$ SQUID system for biological immunoassays

  • Enpuku, K.
    • 한국초전도학회:학술대회논문집
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    • v.10
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    • pp.7-7
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    • 2000
  • A high $T_c$ SQUID system is developed for the application to biological immunoassay. In this application, magnetic nanoparticles are used as magnetic markers to perform immunoassay, i.e., to detect binding reaction between an antigen and its antibody. The antibody is labeled with ${\gamma}-Fe_2O_3\;(or\;Fe_3O_4)$ nanoparticles, and the binding reaction can be magnetically detected by measuring the magnetic field from the nanoparticles. Design and set up of the system is described. The system consists of (1) SQUID magnetometer or gradiometer made of 30-deg. bicrystal junctions, (2) field and compensation coils to apply the magnetic field of about 1 mT, (3) special Dewar to realize a 2 mm-distance between the SQUID and the sample, (4) two layers of cylindrical shielding to reduce the extemal magnetic noise to about 1/100, and (5) an electric slider to move the sample with a speed of 10 mm/sec. The sensitivity of the system is studied in terms of detectable magnetic flux. For the measurement bandwidth from 0.2 Hz to 10 Hz, minimum-detectable amplitude of the magnetic flux is $0.8\;m\;{\Phi}_o$ and $0.25\;m{\Phi}_o$ for the magnetometer and the gradiometer, respectively, when the magnetic field of 1 mT is applied. The difference between them is due to the residual environmental noise, and the applied magnetic field does not increase the system noise. The corresponding weight of the magnetic markers is 1 ng and 310 pg, respectively. An experiment is also conducted to measure antigen-antibody reaction with the present system. It is shown that the sensitivity of the present system is 10 times better than that of the conventional method using an optical marker. A one order of magnitude improvement of sensitivity will be realized by the sophistication of the present system.

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Characterization of Monoclonal Antibodies against Human Leukocyte Common Antigen (CD45)

  • Shin, Hyang-Mi;Cho, Woon-Dong;Lee, Geon-Kook;Lee, Seon-Hwa;Lee, Kyung-Mee;Ji, Gil-Yong;Yoon, Sang-Soon;Koo, Ji-Hae;Lee, Ho-Chang;Lee, Ki-Hyeong;Song, Hyung-Geun
    • IMMUNE NETWORK
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    • v.11 no.2
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    • pp.114-122
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    • 2011
  • Background: The leukocyte common antigen (CD45) is a transmembrane-type protein tyrosine phosphatase that has five isoforms. Methods: We generated seven murine mAbs against human CD45 by injecting cells from different origins, such as human thymocytes, PBMCs, and leukemic cell lines. By using various immunological methods including flow cytometry, immunohistochemistry, and immunoprecipitation, we evaluated the reactivity of those mAbs to CD45 of thymus as well as tonsil lysates. Furthermore, we transiently transfected COS-7 cells with each of gene constructs that express five human CD45 isoforms respectively, and examined the specificities of the mAbs against the transfected isoforms. Results: In case of thymocytes, lymphocytes, and monocytes, all the seven mAbs demonstrated positive reactivities whereas none was reactive to erythrocytes and platelets. The majority of immune cells in formalin-fixed paraffin-embedded thymus and tonsil tissues displayed strong membranous immunoreactivity, and the main antigen was detected near 220 kDa in all cases. Among the mAbs, four mAbs (AP4, DN11, SHL-1, and P6) recognized a region commonly present in all the five isoforms. One mAb, YG27, recognized four isoforms (ABC, AB, BC, and O). Two mAbs, P1 and P14, recognized the isoforms that contain exon A encoded regions (ABC and AB). Conclusion: In this study, we confirmed that AP4, DN11, SHL-1, YG27 and P6, are mAbs reactive with the CD45 antigen whereas P1 and P14 are reactive with the CD45RA antigen.

Expression of Antigen Presenting Function-Associated Surface Molecules on $Interferon{\gamma}$-Treated Gingival Fibroblasts and Periodontal Ligament Fibroblasts (($Interferon{\gamma}$)로 자극된 치은섬유아세포와 치주인대섬유아세 포에서 항원제시기능과 관련된 세포 표면분자의 발현)

  • Seo, Seok-Ran;Ryu, Sung-HunO;Oh, Gwi-Ok;Kim, Hyung-Seop
    • Journal of Periodontal and Implant Science
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    • v.30 no.4
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    • pp.895-913
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    • 2000
  • It is becoming increasingly clear that human gingival fibroblasts(HGF) may play a role in regulating immune responsiveness in inflammatory periodontal lesions. Stimulation of HGF with locally-secreted T cell cytokine $IFN_{\gamma}$ induces human leukocyte antigen class II(HLA II) expression on HGF, which is one of the characteristic feature of professional antigen presenting cells(pAPC). However, $IFN_{\gamma}$-treated HGF and other nonprofessional antigen presenting cells(npAPC) are known to be ineffective or less effective antigen presenter to resting T cells. This study, therefore, was undertaken in an effort to elucidate the differences in expression of cell surface molecules between npAPC in periodontal tissues, such as HGF and periodontal ligament fibroblasts(PDLF), and pAPC such as monocytes/macrophages. Using flow cytometry, the levels of cell surface expression of HLA-D, ICAM-1, LFA-3, and B7-1, which are involved in antigen presentation, were determined in HGF, PDLF and human myelomonocytic cell line THP-1. $IFN_{\gamma}$ clearly induced HLA-D expression on both of fibroblasts and monocytes dose dependently. However, expression level on monocytes were 4 to 5 times higher than that on fibroblasts, and induction rate was faster in monocytes than in fibroblasts. The levels of ICAM-1 expression on fibroblasts and monocytes were enhanced by $IFN_{\gamma}$ in a dose dependent manner. On the other hand, the expression of LFA-3 molecule, which could be detected in fibroblasts and monocytes without cytokine stimulation, was no more enhanced by addition of $IFN_{\gamma}$. B7-1, important costimulatory molecule in T cell activation and proliferation, was not detected on both of fibroblasts and monocytes even when stimulated with $IFN_{\gamma}$, except on monocytes fully differentiated by pretreatment of PMA and treated by $IFN_{\gamma}$. These results suggest that delayed expression of HLA-D and absence of B7-1 on $IFN_{\gamma}$ - treated fibroblasts may at least in part be involved in the ineffectiveness of fibroblasts as primary APC. And it is postulated that although periodontal fibroblasts may not serve as primary APC in normal periodontium, sustained expression of HLA II on ubiquitous fibroblasts in inflammatory lesions may perpetuate immune responses and produce chronic inflammation and tissue injury.

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Increased Immunogenicity and Protective Efficacy of a P. aeruginosa Vaccine in Mice Using an Alum and De-O-Acylated Lipooligosaccharide Adjuvant System

  • Ryu, Ji In;Wui, Seo Ri;Ko, Ara;Do, Hien Thi Thu;Lee, Yeon Jeong;Kim, Hark Jun;Rhee, Inmoo;Park, Shin Ae;Kim, Kwang Sung;Cho, Yang Je;Lee, Na Gyong
    • Journal of Microbiology and Biotechnology
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    • v.27 no.8
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    • pp.1539-1548
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    • 2017
  • Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic pathogen that commonly causes fatal infections in cystic fibrosis and burn patients as well as in patients who are hospitalized or have impaired immune systems. P. aeruginosa infections are difficult to treat owing to the high resistance of the pathogen to conventional antibiotics. Despite several efforts, no effective prophylactic vaccines against P. aeruginosa are currently available. In this study, we investigated the activity of the CIA06 adjuvant system, which is composed of alum and de-O-acylated lipooligosaccharide, on a P. aeruginosa outer membrane protein (OMP) antigen vaccine in mice. The results indicated that CIA06 significantly increased the antigen-specific IgG titers and opsonophagocytic activity of immune sera against P. aeruginosa. In addition, the antibodies induced by the CIA06-adjuvanted vaccine exhibited higher cross-reactivity with heterologous P. aeruginosa strains. Finally, mice immunized with the CIA06-adjuvanted vaccine were effectively protected from lethal P. aeruginosa challenge. Based on these data, we suggest that the CIA06 adjuvant system might be used to promote the immunogenicity and protective efficacy of the P. aeruginosa OMP vaccine.