• Title/Summary/Keyword: Surface antigen

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High Expression of Water-Soluble Recombinant Antigenic Domains of Toxoplasma gondii Secretory Organelles

  • Yang, Zhaoshou;Ahn, Hye-Jin;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.52 no.4
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    • pp.367-376
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    • 2014
  • Recombinant antigenic proteins of Toxoplasma gondii are alternative source of antigens which are easily obtainable for serodiagnosis of toxoplasmosis. In this study, highly antigenic secretory organellar proteins, dense granular GRA2 and GRA3, rhoptrial ROP2, and micronemal MIC2, were analyzed by bioinformatics approach to express as water-soluble forms of antigenic domains. The transmembrane region and disorder tendency of 4 secretory proteins were predicted to clone the genes into pGEX-4T-1 vector. Recombinant plasmids were transformed into BL21 (DE3) pLysS E. coli, and GST fusion proteins were expressed with IPTG. As a result, GST fusion proteins with $GRA2_{25-105}$, $GRA3_{39-138}$, $ROP2_{324-561}$, and $MIC2_{1-284}$ domains had respectively higher value of IgG avidity. The $rGST-GRA2_{25-105}$ and $rGST-GRA3_{39-138}$ were soluble, while $rGST-ROP2_{324-561}$ and $rGST-MIC2_{1-284}$ were not. $GRA2_{31-71}$, intrinsically unstructured domain (IUD) of GRA2, was used as a linker to enhance the solubility. The $rGST-GRA2_{31-71}-ROP2_{324-561}$, a chimeric protein, appeared to be soluble. Moreover, $rGST-GRA2_{31-71}-MIC2_{1-284}$ was also soluble and had higher IgG avidity comparing to $rGST-MIC2_{1-284}$. These 4 highly expressed and water-soluble recombinant antigenic proteins may be promising candidates to improve the serodiagnosis of toxoplasmosis in addition to the major surface antigen of SAG1.

Elimination of Mother-to-Infant Transmission of Hepatitis B Virus: 35 Years of Experience

  • Lu, Fang-Ting;Ni, Yen-Hsuan
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.23 no.4
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    • pp.311-318
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    • 2020
  • Chronic hepatitis B viral (HBV) infection remains a major health threat, especially in high-prevalence areas. Most infants infected by mother-to-infant HBV transmission become chronic carriers. In Taiwan, many important preventive interventions have been implemented to block the perinatal transmission of HBV in the past 35 years. The first nationwide universal HBV vaccination program was launched in Taiwan in July 1984. The three-dose HBV vaccine completion rate reached 98.1% in 2018. The prevalence of Hepatitis B surface antigen (HBsAg) decreased from 9.8% in pre-vaccinated period in 1984 to 0.5% in the vaccinated cohort in 2014. The incidence of hepatocellular carcinoma in children aged 6-9 years significantly declined from 0.52 to 0.13 per 100,000 children born before and after 1984, respectively. Furthermore, we have performed a maternal HBV screening program during pregnancy since 1984, with the screening rate peaked at 93% in 2012. The HBsAg- and HBeAg-seropositive rate in pregnant women declined from 13.4% and 6.4% in 1984-1985 to 5.9% and 1.0% in 2016, respectively. To closely control perinatal HBV infection, we have administered hepatitis B immunoglobulin immediately after birth and checked the serum level of HBsAg and anti-HBs in high-risk babies born to HBsAg-seropositive mothers, irrespective of their HBeAg status, since July 2019. We have also adopted short-term antiviral treatments such as tenofovir 300 mg daily in the third trimester for highly viremic mothers and reduced the perinatal infection rates from 10.7 to 1.5%. Through all these efforts, we expect to meet the global goal of eliminating HBV infection by 2030.

Seroprevalence of antibody to the hepatitis C virus in methamphetamine abusers (Methamphetamin 남용입원환자들의 C형간염 항체 양성률에 관한 조사)

  • Kim, Jin-Kyoo;Lee, Ji-Ho;Cho, Byung-Mann;Lee, Soo-Il
    • Journal of Preventive Medicine and Public Health
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    • v.24 no.4 s.36
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    • pp.465-472
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    • 1991
  • This study was designed to estimate the prevalence of hepatitis C virus(HCV) infection in drug abusers. The subjects were 141 inpatients who had been admitted to a general hospital with the symptoms and signs of methamphetamine intoxication. Seroprevalence of antibody to the hepatitis C virus(anti-HCV) was 60.3%(85/141) and it was higher in the group with increased frequency and duration of drug abuse, but such a relationship was not found in the seroprevalence of hepatitis B surface antigen(HBsAg). These findings suggested the possibility of high prevalence of HCV infection in methamphetamine abusers, and the importance of repetitive percutaneous injection in the transmission of HCV infection.

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Bacterial strains isolated from Jeotgal (salted seafood) induce maturation and cytokine production in mouse bone marrow-derived dendritic cells (마우스 골수 유래 수지상세포의 성숙과 사이토카인 생산에 대한 젓갈 분리균의 효과 연구)

  • Moon, Sun-Young;Park, Eun-Jin;Joo, Hong-Gu
    • Korean Journal of Veterinary Research
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    • v.54 no.3
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    • pp.139-146
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    • 2014
  • Jeotgal (salted seafood) has been one of major fermented foods in Korea for long time. Although there are many studies about Jeotgal in various aspects of food, its immunological importance on hosts has not been elucidated yet. In this study, we investigated if several bacteria isolated from Jeotgal may modulate the function of dendritic cells (DCs), powerful antigen-presenting cells equipped with special immunological capabilities. 4 Jeotgal bacteria were selected as representatives and used for experiments. To treat viable DCs, those bacteria were killed at $60^{\circ}C$ for 30 min. The viability of DCs treated with Jeotgal bacteria was verified and two isolates significantly induced high production of interleukin-12, a representative cell-mediated cytokine of DCs. Surface activation and maturation markers (MHC class II, CD40, CD86) of DCs were analyzed by flow cytometer. In addition, the treated DCs showed significantly high lymphocyte stimulatory capability compared to control DCs based on allogeneic mixed lymphocyte reactions. These observations suggest that Jeotgal isolates can function as immunostimulating bacteria in hosts, like Lactobacillus. Taken together, these experimental evidences may broaden the use of Jeotgal isolates in immunological fields in addition to as a fermented food.

Hepatocellular Carcinoma with Right Atrial Invasion Detected by PET/CT (우심방에 침범한 간세포암을 PET/CT로 진단한 1예)

  • Kim, Ji-Hoon;Kim, Eun-Sil;Yu, Ji-Won;Ahn, Seok-Jin;Jung, Jun-Oh;Kim, So-Yon;Kim, Young-Jung
    • Nuclear Medicine and Molecular Imaging
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    • v.42 no.5
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    • pp.414-418
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    • 2008
  • The role of positron emission tomography (PET) with F-18 fluorodeoxyglucose (F-18 FDG) in the diagnosis of hepatocellulcar carcinoma (HCC) has been limited because of a variable FDG uptake in HCC. However, the usefulness of PET/CT for detecting extrahepatic metastasis and monitoring of the treatment response in HCC has been reported. A 55-year-old man with a hepatitis B surface antigen-positive, was admitted to our hospital due to dyspnea, general weakness and body weight loss for one month. Chest X-ray showed multiple reticulo-nodular densities on both lower lung fields, which implies metastatic lesions. F-18 FDG PET/CT revealed consecutively intense hypermetabolic mass in right hepatic lobe, inferior vena cava and right atrium. We report a case of HCC with IVC and right atrium invasion identified by F-18 FDG PET/CT.

Rapid diagnosis of Pseudotuberclosis in yellowtail (Seriola quinqueradiata) by immunofluorescent antibody technique (면역형광항체법(免疫螢光抗體法)에 의(依)한 방어의 유결절증(類結節症) 신속(迅速) 진단(診斷))

  • Bang, Jong-Deuk;Jung, Sung-Hee;Chun, Seh-Kyu
    • Journal of fish pathology
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    • v.3 no.1
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    • pp.11-19
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    • 1990
  • Yellow tail (Seriola quinqueradiata) infected by Pasteurella piscicida have been occurred to mass mortality without showing apparent surface lesions in cage culture farms. In this case, it is necessary to consider countermeasure by rapid diagnosis of infected fish. The purpose of the present study was to investigate usefulness of the direct fluorescent antibody technique(FAT) for rapid diagnosis of pseudotuberclosis of cultured yellowtail caused by P. piscicida. Antibody produced by inoculating rabbit with formalin killed pseudotuberclosis bacteria antigen(strain KNP-2). Immunoglobulin-G(IgG) was purified from antisera by DEAE-cellulose column chromatography and conjugate with fluorescein isothiocyanate. Fluorescein-labeled antisera was purified by sephadex G-25 gel column chromatography. The fluorescein/protein molar ratio of labeled antisera was determined as 8.8-9.5. Diagnosis of cultured yellowtail was examined in cage culture farms which located in Tongyung, kyungnam from July to October 1990. The causative bacteria of pseudotuberclosis could be detected within two hours after the specimens were transferred to the laboratory for FAT, and it showed that FAT could be adapted as a rapid and accurate diagnostic method of pseudotuberclosis in yellowtail.

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Fiber-optic fluoroimmunosensor for foodborn pathogens using an optical evanescent field (광섬유 소산장을 이용한 식중독균 신속검출용 형광면역센서)

  • Yeom, Se-Hyuk;Park, Chang-Sub;Kim, Do-Eok;Kim, Kyu-Jin;Kang, Byoung-Ho;Kang, Shin-Won
    • Journal of Sensor Science and Technology
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    • v.16 no.6
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    • pp.441-448
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    • 2007
  • In this study, the fiber-optic fluoro-immunosensor was designed to detect foodborne pathogens. The fabricated system is composed of the multimode optical fiber on which antibodies are immobilized. Then, a sandwich immunoassay is applied to the fabricated the fiber-optic fluoro-immunosensor. In the "sandwich" binding format, a primary or "capture" antibody is immobilized on the core surface of the multimode optical fiber and a secondary or named as "tracer" antibody is added to the bulk solution. A tracer is labeled FITC (fluorescein isothiocyanate; ${\lambda}ex$=492 nm, ${\lambda}em$= 520 nm). Different concentrations of antigens are tested in different fibers. The detection limit of the fabricated system is 5.08×103 cfu/ml for Vibrio antigen and $0.1{\mu}g/ml$, $0.05{\mu}g/ml$ in non-labeled monolayer phosphate buffered saline (NMP), non-labeled monolayer carbonate bicarbonate buffer (NMC), respectively.

Analysis of the Molecular Event of ICAM-1 Interaction with LFA-1 During Leukocyte Adhesion Using a Reconstituted Mammalian Cell Expression Model

  • Han, Weon-Cheol;Kim, Kwon-Seop;Park, Jae-Seung;Hwang, Sung-Yeoun;Moon, Hyung-Bae;Chung, Hun-Taeg;Jun, Chang-Duk
    • Animal cells and systems
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    • v.5 no.3
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    • pp.253-262
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    • 2001
  • Ligand-receptor clustering event is the most important step in leukocyte adhesion and spreading on endothelial cells. Intercellular adhesion molecule-1 (ICAM-1) has been shown to enhance leukocyte adhesion, but the molecular event during the process of adhesion is unclear. To visualize the dynamics of ICAM-1 movement during adhesion, we have engineered stable Chinese hamster ovary cell lines expressing ICAM-1 fused to a green fluorescent protein (IC1_GFP/CHO) and examined them under the fluorescence microscopy. The transfection of IC1_GFP alone in these cells was sufficient to support the adhesion of K562 cells that express $\alpha$L$\beta$2 (LFA-1) integrin stimulated by CBR LFA-1/2 mAb. This phenomenon was mediated by ICAM-1-LFA-1 interactions, as an mAb that specifically inhibits ICAM-1-LFA-1 interaction (RRl/l) completely abolished the adhesion of LFA-1* cells to IC1_ GFP/CHO cells. We found that the characteristic of adhesion was followed almost immediately (~10 min) by the rapid accumulation of ICAM-1 on CHO cells at a tight interface between the two cells. Interestingly, ICI_GFP/CHO cells projected plasma membrane and encircled approximately half surface of LFA-1+ cells, as defined by confocal microscopy. This unusual phenomenon was also confirmed on HUVEC after adhesion of LFA-1* cells. Neither cytochalasin D nor 2,3-butanedione 2-monoxime an inhibitor of myosin light chain kinase blocked LFA-1-mediated ICAM-1 clustering, indicating that actin cytoskeleton and myosin-dependent contractility are not necessary for ICAM-1 clustering. Taken together, we suggest that leukocyte adhesion to endothelial cells induces specialized form of ICAM-1 clustering that is distinct from immunological synapse mediated by T cell interaction with antigen presenting cells.

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Prostate Volume Measurement by Transrectal Ultrasonography in the Healthy Korean Men in Thirties (건강한 30대 한국인 남성에서 경직장초음파를 이용한 전립선 용적 측정)

  • Kim, Yon-Min;Jeong, Jeong-Yun;Han, Dong-Kyoon
    • The Journal of the Korea Contents Association
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    • v.12 no.6
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    • pp.338-345
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    • 2012
  • Benign prostatic hyperplasia (BPH) is increasing in Korea owing to the westernized eating habits and changes in living environment. However, very few studies have been made to determine the standard value of the prostate volume as a diagnostic reference. Also, most of the previous studies have been focusing on the elder population over 40 years old. This study retrospectively analyzes the results of prostate volume measured by transrectal ultrasonography in the healthy Korean men in their 30's who visited our institution's center for health promotion for routine checkups. The mean age of the total of 157 men enrolled was $34.6{\pm}2.9$ years, and the mean prostate volume was $19.4{\pm}4.0ml$. Prostate volume of these men was correlates proportionally to their age, prostate specific antigen, body mass index, body surface area, and abdominal girth. Among these, the strongest correlation is between the prostate volume and the abdominal girth (r=0.317, p<0.001).

Receptor-Mediated Endocytosis of Hepatitis B Virus PreS1d Protein in EBV-Transformed B-Cell line

  • Park, Jung-Hyun;Cho, Eun-Wie;Lee, Dong-Gun;Park, Jung-Min;Lee, Yun-Jung;Choi, Eun-A;Kim, Kill-Lyong
    • Journal of Microbiology and Biotechnology
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    • v.10 no.6
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    • pp.844-850
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    • 2000
  • The specific binding and internalization of viral particles is an essential step for the successful infection of viral pathogens. In the case of the hepatitis B virus (HBV), virions bind to the host cell via the preS domain of the viral surface antigen and are subsequently internalized by endocytosis. HBV-preS specific receptors are primarily expressed on hepatocytes, however, viral DNA and proteins have also been detected in extrahepatic sites, suggsting that celluar recepators for HBV may also exist on extrahepatic cells. Recently, an EBV-transformed B-cell line was identified onto which the preS region binds in a receptor-ligand specific manner. In this study, this specific interaction was further characterized, and the binding region within the preS protein was locaized. Also the internalization after host cell attachment was visualized and analyzed by fluorescence-labeled HBV-preS1 proteins using confocal microscopy. Energy depletion by sodium azide treatment effectively inhibited the internalization of the membrane-bound preS1 ligands, thereby indicating an energy-dependent receptor-mediated endocytotic pathway. Accordingly, the interaction of HBV-pres! with this specific B-cell line may serve as an effective model for an infection pathway in extrahepatic cells.

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