• Korean Journal of Veterinary Research
• /
• v.39 no.4
• /
• pp.797-805
• /
• 1999

T sergenti cDNA library were constructed to get a more broad information about the structural, functional or antigenic properties of the proteins, and analyzes for their partial cDNA sequences and expression sequences tags(ESTg). The mRNA were purified from T sergenti isolates to identify the information of antigen gene, then first and second strand cDNA was synthesized. EcoR I adaptor ligation and Xho I enzyme restriction were used to the synthesized cDNA, and ligated into a Uni-ZAP XR vector. T sergenti cDNA library was constructed with packaging and amplification in vitro. Antibody screening was performed with constructed T sergenti cDNA library using antisera against T sergenti. Among those clones, eight phagemids were rescued from the recombinant in vivo excision with f1 helper phage. Using the analysis of endonuclease restriction and PCR, the recombinant cDNA were proved having a 0.5-3.0kb of inserts. The eight of partial cDNA clones' sequences were obtained and examined for their homology using BLASTN and BLASTX. The eight of sequenced clones were classified into three groups according to the basis of database searches. A total 3,045bp of partial cDNA sequence were determined from six clones. The putatively identified clones contain a cytochrome c gene, a heat shock protein gene, a cyclophilin gene, and a ribosomal protein gene. The unidentified clones have a homology to ATP-binding protein(mtrA) gene of S argillaceus, DNA-binding protein(DBP) gene of Pseudorabies virus 85kDa merozoite protein gene of B bovis, mRNA spm1 protein of T annulata and glycine-rich RNA-binding protein mRNA of O sativa etc.

• Journal of Preventive Medicine and Public Health
• /
• v.17 no.1
• /
• pp.223-229
• /
• 1984

An attempt to confirm the associations of some selected risk factors of HBV infection and measure their risks, a cross-sectional study with 1,209 urban office workers was carried out. For the study, a simple questionnaire which contained several questions on personal experience and behaviors on several known selected risk factors of HBV infection was applied to each subject, and the Hepatitis B virus surface antigen and its antibody were checked by RPHA and PHA method, respectively. Risk factors chosen for this study were experience of blood transfusion and personal contact variables, such as frequencies of eating-out, drinking after office hours, going to tea room, sharing cigarettes, etc. The results obtained were as follows: 1. The proportion of HBsAg positive was 10.6%, and total HVB infected including the Anti-HBs positive cases without vaccination was 44.2%. Both were higher in male than in female. 2. Frequent personal contact through glasses and dishes in eating-outs and drinkings turned out not to be a significant risk factor of Hepatitis B surface antigenecity. 3. Frequent visits to tea room was a significant risk factor of HBV infection which combined HBsAg positive cases and Anti-HBs cases who had not received HBV vaccination. The odds ratio was 1.56 4. Blood transfusion was not a significant risk factor of both HBsAg positive and total HBV infection. In summary, indirect oral contacts through eating-outs and drinkings was not significant risk factor in Korea at least between adults. Blood transfusion is no more major source of HBV infection in Korea probably because the adquate screening test of HBsAg for the blood donors is being made.

• Journal of environmental and Sanitary engineering
• /
• v.4 no.1 s.6
• /
• pp.17-28
• /
• 1989

Having studied the production of monoclonal antibodies for developing a diagnosis medicine which shall be detected by a high-sensitivity test by using Rhodotorula rubra as a fungi-host which had been extracted through biochemical tests and follow-up examinations on Yeast-like fungi obtained from pulmonary tissues of pulmonary tuberculosis patients who had been in Kong ju National Tuberculosis Hospital from Jun. to Dec. in 1987, I. have gained such results as follows: 1. The fusion rate was influenced by feeder cell layers, cell density and time required to the cell fusion with cells in myelona subculture. 2. The fusion rate did not show any significant difference when the cell was applyed with two molecular weights, i.e., 1500 and 4000, of polyethylene glycol. 3. Fused cells after the addition of HAT selection media were bright and round, whereas unfused myelona cells and spleen cells were shrunk and granulated. 4. The cell fusion rate turned out to be about $57.2\%$(150 wells / 264 wells). 5. $10\%$(15 wells / 150 wells) of the positive reaction was detected in monoclonal antibody screening. 6. The titer which had reacted positively to Rhodotorula rubra fungal-host was 800 times in density after the gradual dilution of the produced monoclonal antibodies with Indirect ELISA method. 7. The Strongest specific reaction came out after the peroxidase labelled anti-human Immunogobulin had been applyed to Rhodotorula rubra for activating its nature after making drift with Carbonate-bicarbonate buffer (pH 9.6) and drying completely.

• Parasites, Hosts and Diseases
• /
• v.46 no.4
• /
• pp.209-216
• /
• 2008

A monoclonal antibody against Toxoplasma gondii of Tg556 clone (Tg556) blotted a 29 kDa protein, which was localized in the dense granules of tachyzoites and secreted into the parasitophorous vacuolar membrane (PVM) after infection to host cells. A cDNA fragment encoding the protein was obtained by screening a T. gondii cDNA expression library with Tg556, and the full-length was completed by 5'-RACE of 2,086 bp containing an open reading frame (ORF) of 669 bp. The ORF encoded a polypeptide of 222 amino acids homologous to the revised GRA3 but not to the first reported one. The polypeptide has 3 hydrophobic moieties of an N-terminal stop transfer sequence and 2 transmembrane domains (TMD) in posterior half of the sequence, a cytoplasmic localization motif after the second TMD and an endoplasmic reticulum (ER) retrival motif in the C-terminal end, which suggests GRA3 as a type III transmembrane protein. With the ORF of GRA3, yeast two-hybrid assay was performed in HeLa cDNA expression library, which resulted in the interaction of GRA3 with calcium modulating ligand (CAMLG), a type II transmembrane protein of ER. The specific binding of GRA3 and CAMLG was confirmed by glutathione S-transferase (GST) pull-down and immunoprecipitation assays. The localities of fluorescence transfectionally expressed from GRA3 and CAMLG plasmids were overlapped completely in HeLa cell cytoplasm. In immunofluorescence assay, GRA3 and CAMLG were shown to be co-localized in the PVM of host cells. Structural binding of PVM-inserted GRA3 to CAMLG of ER suggested the receptor-ligand of ER recruitment to PVM during the parasitism of T. gondii.

• Pediatric Gastroenterology, Hepatology & Nutrition
• /
• v.26 no.4
• /
• pp.201-212
• /
• 2023

Purpose: The gastrointestinal system is the most commonly affected organ, followed by the lungs, in patients with primary immunodeficiency disease (PID). Hence, it is common for children with PIDs to present with gastrointestinal symptoms. We aimed to analyze the clinical and histopathological findings of patients who were initially admitted to pediatric gastroenterology/hepatology clinics and subsequently diagnosed with PIDs to identify the clinical clues for PIDs. Methods: The demographic, laboratory, and histopathological findings, treatment modality, and outcomes of patients initially admitted to the pediatric gastroenterology/hepatology unit and subsequently diagnosed with PIDs were recorded. Results: The study included 24 patients (58.3% male; median age [range]: 29 [0.5-204] months). Common clinical presentations included chronic diarrhea (n=8), colitis (n=6), acute hepatitis (n=4), and acute liver failure (n=2). The association of autoimmune diseases, development of malignant diseases, and severe progression of viral diseases was observed in 20.8%, 8.3%, and 16.6% of the patients, respectively. Antibody deficiency was predominantly diagnosed in 29.2% of patients, combined immunodeficiency in 20.8%, immune dysregulation in 12.5%, defects in intrinsic and innate immunity in 4.2%, autoinflammatory disorders in 8.3%, and congenital defects of phagocytes in 4.2%. Five patients remained unclassified (20.8%). Conclusion: Patients with PIDs may initially experience gastrointestinal or liver problems. It is recommended that the association of autoimmune or malignant diseases or severe progression of viral diseases provide pediatric gastroenterologists some suspicion of PIDs. After screening using basic laboratory tests, genetic analysis is mandatory for a definitive diagnosis.

• International Journal of Stem Cells
• /
• v.15 no.3
• /
• pp.233-246
• /
• 2022

Background and Objectives: Systemic lupus erythematosus (SLE) is a chronic autoimmune disease mainly affecting young women of childbearing age. SLE affects the skin, joints, muscles, kidneys, lungs, and heart. Cardiovascular complications are common causes of death in patients with SLE. However, the complexity of the cardiovascular system and the rarity of SLE make it difficult to investigate these morbidities. Patient-derived induced pluripotent stem cells (iPSCs) serve as a novel tool for drug screening and pathophysiological studies in the absence of patient samples. Methods and Results: We differentiated CMs from HC- and SLE-iPSCs using 2D culture platforms. SLE-CMs showed decreased proliferation and increased levels of fibrosis and hypertrophy marker expression; however, HC-and SLE-monolayer CMs reacted differently to SLE serum treatment. HC-iPSCs were also differentiated into CMs using 3D spheroid culture and anti-Ro autoantibody was treated along with SLE serum. 3D-HC-CMs generated more mature CMs compared to the CMs generated using 2D culture. The treatment of anti-Ro autoantibody rapidly increased the gene expression of fibrosis, hypertrophy, and apoptosis markers, and altered the calcium signaling in the CMs. Conclusions: iPSC derived cardiomyocytes with patient-derived serum, and anti-Ro antibody treatment could serve in effective autoimmune disease modeling including SLE. We believe that the present study might briefly provide possibilities on the application of a combination of patient-derived materials and iPSCs in disease modeling of autoimmune diseases.

• Journal of Preventive Medicine and Public Health
• /
• v.37 no.4
• /
• pp.337-344
• /
• 2004

Objectives : This study was performed to determine the incidence density and the prevalence of sero-positive hepatitis C from 1999 to 2002 among adults aged 20 and over residing in Seoul and the Gyeonggi province. Method : The data for period was obtained from 114,635 adults, residing in Seoul or the Gyeonggi province, who had undertaken comprehensive health screening tests from Jan 1999 to Dec 2002 in a University hospital in Seoul. Among them, subjects with sero-negative status against hepatitis C were selected (21,408 in 1999, 28,830 in 2000) and then followed up until Dec 2002 to determine the incidence of hepatitis C during this period. The serum was tested with the immunoradiometric assay (IRMA) which uses third generation HCV antibody. Age adjusted rates were estimated by direct standardization using a reference population of 2000 aged from 20 to 80 years. Results : The prevalence of anti-HCV from 1999 to 2002 was 2.1 per 1000 persons(95% CI $1.8{\sim}2.4$). Male showed 1.7 per 1000 persons (95% CI $1.4{\sim}2.1$), while female showed 2.7 per 1000 persons(95% CI $2.2{\sim}3.2$). Age?sex adjusted rate showed 2.8 per 1000 persons (95% CI $2.64{\sim}2.96$), which is lower than the results of some previous study. The prevalence showed a significantly increasing pattern with age both in males and females (p<0.05). The incidence density of anti-HCV among the population aged 20 and over was 1.1 per 104 person-years at risk (95% CI $0.6{\sim}2.4$); 1.2 (95% CI $0.6{\sim}2.7$) for males and 0.8 (95% CI $0.6{\sim}4.2$) for females. Age adjusted incidence density was 2.91 per 104 person-years at risk (95% CI $2.43{\sim}3.38$) for those aged 20 and over. It showed an increasing pattern with age (p<0.05), especially for those age over 50 years. Conclusion : The study subjects for this study were supposedly healthier than the general population so the prevalence and incidence for the general population are thought to be higher than the results of the present study.

• Korean Journal of Food Science and Technology
• /
• v.42 no.3
• /
• pp.329-334
• /
• 2010

Escherichia coli O157:H7 causes hemolytic uremic syndrome and hemorrhagic colitis in humans. The objectives of this study were to produce monoclonal antibody(MAb) against E. coli O157:H7 and to develop an enzyme linked immunosorbent assay(ELISA) for the rapid detection of E. coli O157:H7 in agri-stockbreeding. The characterization of MAb produced from hybridoma cell (HKEC 4G8-5) was validated by ELISA and Western blot. The produced MAb was specific to E. coli O157:H7 and showed weak cross-reaction to Staphylococcus aureus. The detection limit of ELISA based on 4G8-5 MAb was $10^5\;cell/mL$. Although the ELISA could not detect E. coli O157:H7 in the meat and sprout samples inoculated with $1{\times}10^1\;cell$/10 g without enrichment, the same samples after enrichment for 6 hr were confirmed to be positive by ELISA. These results indicated that the ELISA combined with short enrichment (6 hr) is useful tool for rapid screening of E. coli O157:H7 in various samples.

• Pediatric Infection and Vaccine
• /
• v.10 no.2
• /
• pp.200-207
• /
• 2003

Purpose : Malaria is known to have been eradicated for a few decades through persistent national health program in South Korea. However, malaria caused by Plasmodium vivax has started to reappear incidiously among military personnel near to DMZ since 1993. After then, the number of malarial cases have been increased abruptly year by year. We analyzed the children of indigenous malaria who were diagnosed by peripheral blood smear and malarial antibody test with regards to epidemiologic and clinical manifestations. Methods : The study 13 cases below 15 years of age, who were confirmed as vivax malaria during from January 2000 to August 2003. We retrospectively analyzed epidemiologic data, clinical manifestations, laboratory findings and therapeutic responses. Results : All of 13 cases were indigenous and tested positive for Plasmodium vivax. Of 13 patients, 9 were male and 4 were female. Mean age of onset was $9.5{\pm}3.6$ years old. Ilsan(n=9) was the most prevalent area, the most patients(n=11) were presented in summer (from June to August). A characteristic feature of periodic 3 day fever in patients with P.vivax infection was reported in only 2 among 13 cases. Thrombocytopenia was most prominent findings, which was accompanied by 12 of 13 patients and pancytopenia was appeared in 3 patients on this study. The therapeutic responses of hydroxycholoquine were very good in all cases, and abnormal laboratory findings were recovered and no relapse during follow-up period. Conclusion : Vivax malaria is indigenous in Korea near to DMZ, but recently endemic area seemed to be extended southward. Plasmodium vivax is the cause of indigenous malaria of children. As for children with high fever accompanying thrombocytopenia in endemic area of Korea, malaria must be included in differential diagnosis whether the type of fewer is periodic 3 day fever or not. Malaria antibody test is helpful as a screening test for malaria.

• Biomedical Science Letters
• /
• v.2 no.1
• /
• pp.41-48
• /
• 1996

The C-reactive protein(CRP) from ascitic and pleural fluid was purified using calcium dependent affinity chromatography of CNBr activated Sepharose-4B covalently coupled to p-diazonium phenylphosphorylcholine(DPPC) and hydroxylapitite chromatography. Polyclonal antibody was prepared from rabbit by immunizing the purified CRP. Specific immunoglobulin G was isolated using affinity chromatography and coupled to microparticles. A sensitive microparticle-based immunoassay was developed to measure CRP within 3 mins. The detection range was between 0.5mg/dl) and 20mg/dl in serum, showing strong response in the range of 0.7~2.9 mg/dl, weak response in 5.0~13.2 mg/dl and zone phenomenon over 28mg/dl. The average value of CRP in 74 samples was 3.8mg/dl and most of the values were lower than 10mg/dl .The CRP values of serum samples were determined by our microparticle-based immunoassay, and were compared with those obtained using the other commercial products(B Co., France and I Co., Japan). Good correlations were shown between the values obtained by our developed microparticle-based immunoassay system and those by other commercial products. All performance characteristics evaluated make our developed microparticles-based immunoassay suitable for a simple, rapid, and reliable screening of CRP in serum.