• Journal of Plant Biotechnology
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• v.29 no.4
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• pp.287-293
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• 2002

We are trying to develop a transgenic carrot with aims of production and delivery of oral vaccine against microbial enteropathogen using a K88ac pilin gene. A K88ac antigen (pilin) gene was isolated by PCR from the K88ac genomic DNA. The pilin gene was constructed in pGA748 and introduced via Agrobacterium tumefaciens to the explants of carrot hypocotyl and then 494 transgenic lines were established. The amounts of the K88ac antigen produced in each of the cell lines were determined by western and two elite cell lines (M1-17, Y14-1) were selected based on higher levels of expression of the antigens as well as rate of cell growth and efficiency of embryogenesis. In order to test an immunization induced by oral administration of the transgenic carrot, serum of the mice fed with the carrot vaccine were tested in ELISA. It tumed out that the mice fed with 3 g of transgenic carrot showed a similar level of antibody compared to those applied with 10 $\mu\textrm{g}$ of the purified recombinant pilin protein. Besides, various clinical responses were measured after challenging with ETEC K88ac strain to the piglets experiencing an oral immunization with the transgenic carrot. The piglets fed with carrot vaccine showed a lower level of diarrhea in fecal score compared to those fed with non-transgenic carrot. A higher level of increase in weight of the piglets fed with the transgenic carrot vaccine was observed comparing to those fed with non-transgenic carrot as control.

• Korean Journal of Veterinary Research
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• v.14 no.1
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• pp.59-71
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• 1974

The studies were conducted on the allergic skin reaction of bovine eurytremiasis. In order to confirm the appearance of allergic skin reaction in this disease, a protein fraction of Eurytrema SPP(EPF) was extracted, and it's antigenicity was studied by means of intradermal reaction on the rabbits, goats and bovine. Rabbits used in this studies were divided into three groups: the first group was sensitized with EPF, the second group was sensitized with EPF, and the third group was served as control which is consisted with 5 non-sensitized and 2 saline injected rabbits. Three healthy native goats and bovine also used. Among these, 86 cases were eurytremiasis, 31 cases were fascioliasis and 71 cases non-infected. A protein fraction was also made from adult worm of Fasciola SPP(FPF) and tested the cross reaction between these two different fluke infestations. The results obtained were as follow: 1. Intradermal reaction was appeared in the infected cattle(bovine eurytremiasis) as well as in the EPF sensitized rabbits and goats. 2. In the EPF sensitized rabbits, the wheal of the intradermal reaction reacted the maximum size (increased wheal size of 5.0 to 9.0 mm) in 40 to 60 minutes and erythema in 60 minutes after intradermal injection of EPF antigen. In the EPF sensitized goats, the size of wheal reacted the maximum(increased wheal size of 5.5 to 8.0 mm) in 30 to 40 minutes, and then disappeared in 120 to 180 minutes after injection of EPF antgen. On the contrary, in the control groups of rabbits and goats, the wheal (7.0 mm) produced following antigen injection reached only to 7.5 to 8.5 mm. in 10 minutes without developing erythema, and then perished in 40 to 60 minutes after injection of EPF antigen. In the infected cattle, the wheal of the intradermal reaction reached the maximal sizes(increased wheal size of 4.0 to 11.0 mm) in 15 to 20 minutes and perished in 60 to 90 minutes after injection of EPF antigen. 3. It would be standardized that the positive reaction is above 4.0 mm, the negative below 2.0 mm and the suspective is 3.0 mm of the infected cattle. In the results of intradermal reaction, 87.2%(75 of 86 cases) was positive, 4.7%(4 of 86 cases) negative and 8.1% (7 of 86 cases) was suspective. However, in the non-infected control cattle, the wheal produced following injection disappeared in 30 to 40 minutes without increasing the wheal size in the most subjects or with incresing only to 1.0 to 2.0 mm in a few cattle. 4. In the results of intradermal reaction which were reciprocally tested in EPF sensitized rabbits by FPF antigen and in FPF sensitized. rabbits by EPF antigen, there were cross reacted. In the 31 cases if fascioliasis tested by EPF antigen, 11 cases were not specifically reacted with EPF antigen. Judging from the results of cross reaction in experimental rabbits and natural bovine fascioliasis, it would be expected when both EPF and EPF antigens are given intracutaneously on the same bovine patients of eurytremiasis or fascioliasis, the former antigen will produce the larger wheal in eurytremiasis, while the latter antigen will produce also the larger wheal in fascioliasis.

• The Korean Journal of Zoology
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• v.31 no.1
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• pp.49-55
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• 1988

A few SV4O-transformed human cells such as SV8O are potentially tumorigenic but rejected by athymic hosts. However, one cell line in this group (W118IVA-2) is known to be fully tumorigenic. Two clones were obtained after the injection of W118IVA-2, of which NW1SC1-1 was tumorigenic but NW18C1-2 was not in nude mice. As examined by Southern blot analysis, NW18C1-1 appears to contain more copy number of SV40 sequences than NW18C1-2 does. However, it was unable to demonstrate that this difference elicits the tumorigenicity in NW18C1-1 but not in NW18C1-2. Therefore, the latter clone was tested if it expresses SV40 early genes to produce large T as well as small t antigens using indirect immunofluorescent assay and immunoprecipitation. In addition, mouse NIH3T3 cells were transfected with the cellular DNA of NW1SC1-2 as well as that of NW18C1-1 to examine if the viral genomes in the clones can make the nontransformed cells to acquire malignant growth potential in vivo. The transformed cells expressed large T antigen and became tumorigenic. Thus, the transforming functions of NW1SC1-2 cell appers to be intact. These results clearly suggest that the inability of NW18C1-2 cell to form tumor in nude mice is not because they are inherently nontumorigenic. However, the possibility that the interaction of SV40 with its host differs in these clones can not he ruled out.

• Journal of Life Science
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• v.24 no.9
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• pp.995-1000
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• 2014

The studies of marine viruses in terms of viral isolation and detection have been limited due to the high mutation rate and genetic diversity of marine viruses. Of the modern methods currently used to detect marine viruses, serological methods based on enzyme-linked immunosorbent assay (ELISA) are the most common. They depend largely on the quality of the antibodies and on highly purified suitable antigens. Recently, a new experimental system for using viral capsid protein as an antigen has been developed using the yeast surface display (YSD) technique. In the present study, the capsid protein gene of the red-spotted grouper nervous necrosis virus (RGNNV) was expressed and purified via YSD and HA-tagging systems, respectively. Two regions of the RGNNV capsid protein gene, RGNNV1 and RGNNV2, were individually synthesized and subcloned into a yeast expression vector, pCTCON. The expressions of each RGNNV capsid protein in the Saccharomyces cerevisiae strain EBY100 were indirectly detected by flow cytometry with fluorescently labeled antibodies, while recognizing the C-terminal c-myc tags encoded by the display vector. The expressed RGNNV capsid proteins were isolated from the yeast surface through the cleavage of the disulfide bond between the Aga1 and Aga2 proteins after ${\beta}$-mercaptoethanol treatment, and they were directly detected by Western blot using anti-HA antibody. These results indicated that YSD and HA-tagging systems could be applicable to the expressions and purification of recombinant RGNNV capsid proteins.

• Journal of Veterinary Clinics
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• v.32 no.6
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• pp.544-547
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• 2015

An 11-year-old, intact male Pekingese was brought to the Veterinary Teaching Hospital of Kangwon National University with a 10-day history of seizures. Fifteen days before coming to Kangwon National University, the dog had visited a local animal hospital for lameness, and non-steroidal anti-inflammatory drugs were prescribed to treat this symptom. However, 10 days before coming to our hospital, the dog experienced generalized seizures. Two days before his arrival, generalized ataxia and mental dullness also occurred. Our examinations revealed no remarkable findings on a routine blood test or X-ray. However, the neurological examinations confirmed mental dullness, generalized ataxia, and a lack of menace response and pupillary light reflexes. Nine hours later, dyspnea occurred, and 12 hours after that, the patient was euthanized per the client's request. A necropsy of transverse sections confirmed the presence of a prominent midline shift due to extended tumor growth. On histopathological analyses, pseudopalisading necrosis of the glial cells and microvascular proliferation were observed. In immunohistochemical analysis, glial fibrillary acidic protein, proliferating cell nuclear antigens, and ionized calcium binding adaptor molecule 1 immunoreactive cells were observed in the tumor area. Based on the results, the tumor was confirmed to be a glioblastoma. Primary intracranial tumors are rare in the veterinary field. This case report describes the clinical and histopathological findings of glioblastoma in a Pekingese.

• The Journal of the Korean Society for Microbiology
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• v.22 no.3
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• pp.309-321
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• 1987

For the investigation of microbiological and immunological specificity of Bacteroides gingivalis, Bacteroides gingivalis were isolated, enumerated and characterized from 13 Korean rapidly progressive periodontitis and 7 healthy control by anaerobic culture technique. The total proportion of black-pigmented Bacteroides from Korean R.P.P. patients and healthy control were 8.78% and 0.92%, respectively, among total isolated black-pigmented Bacteroides. In antibiotic susceptibility test, Bacteroides gingivalis isolated from R.P.P. patients were sensitive to Ampicillin and Tetracycline, and resistant to Gentamicin and Erythromycin in disc diffusion method. In antibiotic broth dilution method, the minimum inhibitory concentration(MIC) to Bacteroides gingivalis was 2 unit/ml of Penicillin and $0.25{\sim}1{\mu}g/ml$ of Tetracycline, respectively. The concentration of serum IgG in rapidly progressive periodontitis patients were sigificantly higher than that of healthy control, and concentration of diluted gingival crevicular IgG has not any significant differences between two groups. Serum and gingival crevicular IgG antibody to Bacteroides gingivalis were significantly higher titer in rapidly progressive periodontitis patients to compare with healthy control. The lipopolysaccharide profiles of 2 Korean B. gingivalis in silver stained sodium dodecyl sulfate-polyacrylamide gel electrophoresis were similar to type strains of B. gingivalis and typical LPS band were appeared around the 24-Kd molecular weight. Immunodiffusion test and immunoelectrophoresis of the L.P.S. extracted from 2 Korean B. gingivalis and 2 kinds of type strains of B. gingivalis showed that B. gingivalis Korean-1 was reacted identically to B. gingivalis ATCC 33277. In trypsin and ${\alpha}$-glucosidase activity test of 2 Korean B. gingivalis, both of them revealed positive trypsin and negative ${\alpha}$-glucosidase activity, respectively. These investigation suggested that B. gingivalis is important pathogenic plaque bacteria for the pathogenesis of periodontitis and further study is needed to purify and characterize of the species-specific antigens of this organisms to develop monoclonal antibody and potential diagnostic reagents.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.12 no.1
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• pp.23-29
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• 2009

Purpose: To provide the primary data for reaching a consensus on the adequate duration of treatment of lamivudine in children with HBeAg negative chronic hepatitis B. Methods: Seven of 83 children/adolescents with chronic hepatitis B were diagnosed with HBeAg-negative and HBV DNA-positive chronic hepatitis B and treated with lamivudine. Six children/adolescents were enrolled among 7 patients, who had been treated with lamivudine over 2 years. The primary goal of treatment was HBV DNA clearance and normalization of the serum ALT level; the final goal of treatment was the durability of the complete response after discontinuation of lamivudine. It was planned to continue lamivudine for more than two additional years after HBV DNA negativity and normalization of ALT. Results: The mean duration of lamivudine treatment was 32.2 months (range, 26~40 months) and the mean duration of follow-up was 59.5 months (range, 26~110 months). HBV DNA levels became undetectable (<0.5 pg/mL) in 6 patients within 3 months of treatment. ALT levels were normalized in 3.5 months (range, 2~7 months) in all 6 patients. Biochemical breakthrough developed in 1 patient 18 months after the initiation of lamivudine treatment. No evidence of relapse could be found in 4 patients with a mean follow-up of 23.8 months (range, 4~75 months) after cessation of lamivudine treatment. Conclusion: Suppression of HBV replication and normalization of serum ALT levels were effectively achieved with long-term lamivudine treatment in children/adolescents with HBeAg-negative chronic hepatitis B. Two additional years of lamivudine may be needed after HBV DNA clearance and ALT normalization in HBeAg-negative chronic hepatitis B in order to decrease the relapse rate.

• Tuberculosis and Respiratory Diseases
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• v.56 no.2
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• pp.159-168
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• 2004

Background : In recent years, numerous human tumor specific antigens such as melanoma antigen gene(MAGE) that is recognized by autologous cytotoxic T lymphocytes have been identified. MAGE is expressed in many human malignancies in various organs, such as lung, breast, stomach, esophagus and leukemia. Therefore MAGE has been studied widely for tumor diagnosis and immunotherapy. But, so far there were no clinical studies evaluating the role of MAGE in pleural effusion. We investigated the expression of MAGE in the patients with exudative pleural effusion for it's diagnostic utility and the results were compared with those of cytologic examinations. Methods : Diagnostic thoracentesis was performed in 44 consecutive patients with exudative pleural effusion during 6 months. We examined the expression of MAGE and cytology with the obtained pleural effusion. Expression of MAGE was interpreted by means of a commercial kit using RT-PCR method. Enrolled patients were divided into two groups such as malignant and benign and we analyzed its' sensitivity and specificity. Results : There were no significant differences between two groups in age, sex, white blood cell counts in pleural fluid, pleural fluid/serum protein ratio and pleural fluid/serum LDH ratio. The sensitivity and specificity of MAGE were 72.2% and 96.2% respectively and the positive predictive value and negative predictive value of MAGE were also 92.9% and 83.3% respectively. The sensitivity and negative predictive value of cytologic examinations were 66.7% and 81.3% respectively. There were no significant differences between sensitivities of MAGE and cytologic examinations but false positive result of MAGE was found in 1 case of tuberculous pleurisy. Conclusion : MAGE is a sensitive and specific marker for the differential diagnosis between benign and malignant effusion in patients with exudative pleural effusion. And MAGE would provide the equal sensitivity compared with that of cytologic examination in patients with malignant pleural effusion if 5mL of the pleural fluid is examined.

• Journal of Korea Association of Health Promotion
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• v.3 no.1
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• pp.72-83
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• 2005

The present pilot project was executed to recommend a strategy of clonorchiasis control in China. The pilot area of this project was Zhaoyuan, Hailin, and Ningan, Heiloagjiang province. A baseline survey subjecting 4,865 residents in Heilongjiang confirmed Zhaoyuan asa high endemic area and Hailin and Ningan as moderate endemic areas. Six different control strategies were implemented in Zhaoyuan, two were in Hailin, and one was in Ningan. Including the baseline survey and project programs from 2000 to 2004, total 63,274subject-times were examined of their feces for Clonorchiseggs, 26,680 were treated, 10,082 were screened by ELISA, and 6,130 subjects were examined of their liver by sonography. The egg Positive rates in 6 villages of Zhaoyuan were as high as 44.8% 70,0%. Following the protocolof each strategy, the subjected residents were examined of their feces and treated with 25 mg/kg praziquantel, 3 times. Except the control group, all of the villages showed 72.8% to 92.0% reduction of their original egg Positive rates at Zhaoyuan. Mass treatments of all subjected residents in 2001 and 2003 reduced the egg rate from 68.8% to18.7% and 4 annual mass treatments reduced the rate from 44.8% in 2001 to 8.7% in 2004.Selective annual treatments of egg positive subjects reduced the egg rates from 50.8% in2001 to 13.8% in 2004 or from 70.0% in 2001 to 11.6% in 2004, and two treatments in a year reduced the rate from 57.6% in 2001 to 4.6% in 2004. According to repeated treatments, EPG counts decreased remarkably. In moderate endemic areas, the original egg rates were 22.6% and 28.3% in 2001 but were 1.7% and 1.1% after 2 or 3 selective treatments. The present findings of the chemotherapeutic control of clonorchiasis prove that repeated medication is important. The reduction is directly correlated with dose of praziquantel but not with mass or selective treatments. Chemotherapeutic control of reservoirhosts has little effect on reinfection of clonorchiasis because the field along the Songhua-jiang is too wide to be impacted. ELISA confirmed many serologically positive cases to Clonorchisantigen but only a few cases were positive to other antigens (Paragonimus, cysticercus, sparganum). The abdominal soaography visualized intrahepatic bile duct dilatation and periductal echo in 2,002 of 6,070 examined subjects. In addition to these examinations and treatment, health education supplemented tㅗe control activities. The present findings prove clonorchiasis is very widely prevalent and heavily endemic along the rivers in Heiloagjiang. The results suggest that group chemotherapy with praziquantel is effective to reduce endemicity of clonorchiasis. Mass treatment without individual fecal examination is recommended in heavy endemic areas where the egg rate is over 40% while one selective treatment is effective enough in moderate endemic areas.

• Laboratory Medicine and Quality Assurance
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• v.40 no.2
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• pp.51-69
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• 2018

As part of the immunoserology program of the Korean Association of External Quality Assessment Service, we organized two trials on the external quality assessment of hepatitis viral markers in 2016 and 2017. The hepatitis viral antigens and antibodies program consisted of 10 test items. We delivered two and three types of pooled sera specimens to 965 and 965 institutions for the first and second trials of external proficiency testing in 2016, respectively. The number of participating laboratories was 915 (94.8%) and 913 (95.0%) in the first and second trials in 2016, respectively. We also delivered three kinds of pooled sera specimens to 936 and 1,015 institutions for the first and second trials of external proficiency testing in 2017, respectively. The number of participating laboratories was 920 (98.3%) and 996 (98.1%) in the first and second trials in 2017, respectively. The most commonly tested items were hepatitis B surface antigen, followed by the antibodies to hepatitis B surface antigen, anti-hepatitis C virus, hepatitis B envelope antigen, antibodies to hepatitis B envelope antigen, anti-hepatitis A virus and antibodies to hepatitis B core antigen. The most frequently used methods for detecting viral markers were the chemiluminescence immunoassay and the electrochemiluminescence immunoassay, but they yielded a few-false positive results due to the matrix effect. The immunochromatographic assay yielded false-negative results for anti-hepatitis A virus due to low sensitivity. Continuous improvement in the quality of viral hepatitis testing through participation in the survey seems necessary.