• Proceedings of the Korean Society of Tobacco Science Conference
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• 1997.10a
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• pp.134-152
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• 1997

Plant viruses of tobacco including tobacco mosaic virus (TMV) and potato virus Y (PVY) cause severe economic losses in leaf-tobacco production. Cultural practices do not provide sufficient control against the viruses. Use of valuable resistant cultivars is most recommendable for the control of the viruses. However, conventional breeding programs are not always proper for the development of virus-resistant plants mostly owing to the frequent lack of genetic sources and introduction of their unwanted properties. Therefore, we tried to develop virus-resistant tobacco plants by transforming commercial tobacco cultivars, NC 82 and Burley 21, with coat protein (CP) or replicase (Nlb) genes of TMV and PVY necrosis strain (PVY-VN) with or without untranslated region (UTR) and with or without mutation. Each cDNA was cloned and inserted in plant expression vectors with 1 or 2 CaMV 35S promotors, and introduced into tobacco leaf tissues by Agrobacterium tumefaciens LBA 4404. Plants were regenerated in kanamycin-containing MS media. Regenerated plants were tested for resistance to TMV and PVY In these studies, we could obtain a TMV-resistant transgenic line transformed with TMV CP and 6 genetic lines with PVY-VN cDNAs out of 8 CP and replicase genes. In this presentation, resistance rates, verification of gene introduction in resistant plants, stability of resistance through generations, characteristics of viral multiplication and translocation in resistant plants, and resistance responses relative to inoculum potential and to various PVY strains will be shown. Yield and quality of leaf tobacco of a promising resistant tobacco line will be presented.

• Korean Journal of Food Science and Technology
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• v.26 no.4
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• pp.436-441
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• 1994

To investigate the lowering effects of in vitro enzymatic hydrolysis by the treatment of chymotrypsin, trypsin, pancreatin, or protease from Aspergillus oryzae on the antigenicity of whey protein isolate (WPI) against rabbit anti ${\alpha}-LA$ antiserum, competitive inhibition ELISA (cELISA) and passive cutaneous anaphylaxis (PCA) test using guinea pig were performed. The results of cELISA showed that the monovalent antigenicity of the whey protein hydrolysates (WPH) to the antiserum was decreased to $10^{-2.5}-10^{-5.5}$ and less by the hydrolysis. The monovalent antigenicity of the WPH hydrolyzed by trypsin, or protease from Asp. nryzae was much lowered by the pretreatment of heat denaturation. The antigenicity of the WPH hydrolyzed by chymotrypsin, trypsin, or pancreatin was much lowered by the pretreatment of pepsin. Especially, the antigenicity of TDP (trypic hydrolysate with pretreatment of heat and pepsin) was found almost to be removed. However, there was not consistency between degree of hydrolysis(DH) and the monovalent antigenicity of the WPH. By the heterologous PCA it was found that all of the PGPH lost the polyvalent antigenicity regardless of the pretreatments although WPI and ${\alpha}-LA$ had the positive high antigenicity. The results suggested that the peptides derived from ${\alpha}-LA$ in WPH could bind specific antibodies but they could not induce allergy. Therefore, it was elucidated that the allergenicity of ${\alpha}-LA$ in whey protein could be destroyed easily by the enzymatic hydrolysis.

• Journal of fish pathology
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• v.6 no.1
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• pp.11-20
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• 1993

To study the immune responses of the japanese eel. Anguilla japonica, fish were injected intraperitoneally with several types of Edwardsiella tarda antigen, i. e., FKC(formalin killed cells), HKC(heat killed cells) or LPS(lipopolysaccharide), and the changes of immunocytes numbers, phagocytosis and agglutination titre in the peripheral blood of the fish were investigated. The number of lymphocytes in the peripheral blood of eels were decreased until 6 hours after injection, and then were turn to normal levels after 24 hours of injection. However, the level were slightly increased and were remained after 24 hours. The number of neutrophils of FKC, HKC or LPS injected fish were the highest at 12 hours after injection and were decreased slowly after that. Three weeks after the injections, the agglutination of antibody titre of all immunized groups were reached at 128 and were remained this level thereafter. However 6 weeks after the injections, that in HKC injected fish were dropped the level up to 4. Fish were injected with LPS and the blood from the fish were bled after 12 hours. Then the blood were incubated with E. tarda. Six hours after incubation, the phagocytic index was reached the highest level, 28.3. One week after the LPS injection, the blood were again bled and incubated with E. tarda. The phagocytic index at this time was 3.9. The phagocytic indexes of the fish injected with FKC and HKC, treted as same LPS injected fish as above, were 18.8 and 10.7, respectively. The phagocytic index of the control fish was 1.2. The antibacterial activities of normal antiserum against E. tarda were shown for both FKC and LPS injected fish, but not for HKC injected fish. The RPS(relative percentage of survival) of HKC, FKC and LPS injected fish in the challenge test were 10%, 20% and 30%, respectively. These results suggest that the effect of protection of the eel which were injected with antigen were varied with the method of preparation of the antigen.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.6 no.2
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• pp.103-111
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• 2003

Purpose: This study was undertaken to evaluate the gastroduodenal pathology and Helicobacter pylori infection in children with upper gastrointestinal symptoms. Methods: One hundred and seven pediatric patients with upper gastrointestinal symptoms were undergone endoscopy at the Gyeongsang National University Hospital from June 1990 to April 1991. Histopathologic examination was done by H & E staining of gastric antral biopsy specimen and gastritis was defined according to the Sydney System. Tissue H. pylori status was evaluated with the urease test using Christensen's urea broth and H & E or Warthin-Starry silver staining of gastric antral biopsy specimen. IgG Immunoblotting were also performed to detect specific anti-H. pylori antibody in these patients. Results: The reasons for endoscopy were recurrent abdominal pain, acute abdominal pain, sallow face, hunger pain, and frequent nausea. Variable degrees of gastric mucosal hyperemia were found in most of the patients. Gastric hemorrhagic spots, gastric ulcer, duodenal ulcer, duodenal erosion, and hemorrhagic duodenitis were rare endoscopic findings. Histologic chronic gastritis was found in 88% of 107 patients. Histologic chronic duodenitis was observed in all 99 patients whose tissue were available. Gastric tissue H. pylori was positive in 57% of 107 patients by one of the ureasetest, H & E staining and Warthin-Starry silver staining. However, gastric tissue H. pylori detection rate was lower in the younger age groups. Anti-H. pylori IgG antibodies were detectable in 96% of 107 patients. Conclusion: Chronic gastroduodenitis and anti-H. pylori IgG antibody were ubiquitous in children with upper gastrointestinal symptoms.

• The Korean Journal of Nuclear Medicine
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• v.32 no.4
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• pp.354-364
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• 1998

Purpose: Simple X-ray study and bone scan have limitations for early diagnosis of bone or bone marrow lesions in multiple myeloma. The purpose of this study was to evaluate the diagnostic usefulness of bone marrow immunoscintigraphy using anti-granulocyte monoclonal antibody for the evaluation of bone involvement in multiple myeloma. Materials and Methods: In 22 patients (Male: 15, Female: 7) with multiple myeloma, we performed whole-body immunoscintigraphy using $^{99m}Tc$-labelled antigranulocyte antibody (BW 250/183, Scintimum $Granulozyt^{(R)}$ CIS, France) and compared the findings with those of simple bone radiography and $^{99m}Tc$-MDP bone scan. Abnormal findings in bone marrow scintigraphy were, considered to be present in case of expansion of peripheral bone marrow or focal photon defect in axial bones. Results: Marrow expansion was noted in 15 of 22 patients (68%). Focal photon defects were found in 18 patients (82%). While one (33%) of 3 patients with Stage II disease showed focal defects in bone marrow scan, abnormal focal defects were observed in 17 of 19 (90%) patients with Stage III. Among 124 focal abnormal sites which were observed in bone marrow scan, bone scan or simple bone radiography, bone marrow scan detected 92 sites (74%), whereas 82 sites (66%) were observed in simple bone radiography(58 sites, 47%) or bone scan(40 sites, 32%). Fifty-one (41%) out of 124 bone lesions were detected by bone marrow scan only, and located mostly in thoracolumbar spine. Conclusion: Bone marrow scan using $^{99m}Tc$-labelled antigranulocyte antibody seems to be a more sensitive procedure for the detection of pathologic bone lesions than simple bone X-ray or bone scan in patients with multiple myeloma.

• Tuberculosis and Respiratory Diseases
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• v.47 no.3
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• pp.311-320
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• 1999

Background: In recent years, tuberculosis has re-emerged as a major health problem in both industrialized & developing countries. Recent advances in identifying & purifying antigens secreted in active tuberculosis infection have lead to the development of serological assays based on a number of immunodominant antigens. To date, the most sensitive and specific of these antigens has been the 38-kDa antigen. Method: Two rapid membrane-based serologic assays using antigen(38-kDa) from mycobacterium tuberculosis for the diagnosis of tuberculosis were evaluated in 22 patients with smear-positive pulmonary tuberculosis, 14 patients with inactive pulmonary tuberculosis, and 9 patients with non-tuberculous lung disease. Result: The evaluation of validity(sensitivity, specificity, positive predictive value, negative predictive value, false positivity and false negativity) of STAT-PAK ULTRA FAST$^{(R)}$ were 77.3%, 28.6%, 63.0%, 44.4%, 71.4 %, and 22.7% for differential diagnosis of active pulmonary tuberculosis and inactive pulmonary tuberculosis, respectively. The evaluation of validity of STAT-PAK ULTRA FAST$^{(R)}$ were 77.3%, 33.3%, 73.9%, 37.5%, 66.7%, and 22.7% for differential diagnosis of active pulmonary tuberculosis and non-tuberculosis. The evaluation of validity of ICT Tuberculosis$^{(R)}$ were 54.5%, 57%, 66.7%, 44.4%, 42.9%, and 45.5% for differential diagnosis of active pulmonary tuberculosis and inactive pulmonary tuberculosis. The evaluation of validity of ICT Tuberculosis$^{(R)}$ were 54.5%, 100%, 100%, 47.4%, 0%, and 45.4% for differential diagnosis of active pulmonary tuberculosis and non-tuberculosis. Conclusion: We concluded no effectiveness of STAT-PAK ULTRA FAST$^{(R)}$ & ICT tuberculosis$^{(R)}$on serologic diagnosis of pulmonary tuberculosis. In the future, further large-scale study should be needed for serologic diagnosis of pulmonary tuberculosis.

• Parasites, Hosts and Diseases
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• v.21 no.2
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• pp.270-280
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• 1983

Agar-gel Diffusion test (AGD), Counterimmunoelectrophoresis(CIEP) and Enzyme-Linked Immunosorbent Assay(ELISA) were examined with the sera of skin test positives for paragonimiasis. The crude antigen (Paragcnimus whole worm extracts: protein concentration, 7.56mg/m1) and human sera were used in AGD and CIEP. And in ELISA test, diluted antigen with 1:40, 000 of crude antigen and diluted sera with 1:100, 1:200 were used in the test. The positive identical ratio between AGD and CIEP reactions is 985 and negative identical ratio is 100%. One or three precipitin bands are observed in AGD. One to seven precipitin bands are also revealed in CIEP. Especially, deeply stained bands are observed in CIEP than those of AGD. The positive identical ratios between AGD and ELISA tests are 96% in 1:100 diluted sera, and 94% in 1:200 diluted sera. But the negative identical ratios between AGD and ELISA tests are 97% and 99% respectively in 1:100 and 1:200 diluted sera. The positive identical ratios between CIEP and ELIEA tests are 98% and 96% respectively in 1:100 and 1:200 diluted sera, but also 97% and 99% in 1:100 and 1:200. Control sera, such as clonorchiasis, amoebiasis and toxoplasmcsis, revealed all negatives with Paragonimus antigen in AGD, CIEP and ELISA tests. By above results, ELISA was mcst sensitive, next CIEP and AGD, But AGD test apprars to be more useful when used to crude antigen without cross rfacticn with other parasitic infections. CIEP test is basically equal in terms of precipitin reaction, but CIEP is able to be detected more sensitively and rapidly though less simple in handiwork than AGD. Consequently, three methods for inlmunological tests of paragonimiasis have gccd correlations with one another. Also, each of these has both merits and demerits in iymunolcgical test for paragonimiasis. But the ELISA test was proved to be the most sensitive and convenient tool for mass screening test, especially in cacti of using purified antigen.

• Journal of Life Science
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• v.26 no.12
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• pp.1383-1391
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• 2016

Anti-estrogen drugs such as tamoxifen have been used for treating patients with ER-positive, early breast cancer. However, resistance to anti-estrogen treatment is inevitable in most patients. Breast cancer anti-estrogen resistance-3 (BCAR3) has been identified as the protein responsible for the induction of tamoxifen resistance in estrogen-dependent human breast cancer. We have previously reported that BCAR3 regulates the cell cycle progression and the signaling pathway of EGF and insulin leading to DNA synthesis. In this study, we investigated the functional role of BCAR3 in regulating c-Jun transcription in non-tumorigenic human breast epithelial MCF-12A cells. A transient transfection of BCAR3 increased both the mRNA and protein of c-Jun expression, and stable expression of BCAR3 increased c-Jun protein expression. The overexpression of BCAR3 directly activated the promoter of c-jun, AP-1, and SRE but not that of $NF-{\kappa}B$. Furthermore, single-cell microinjection of BCAR3 expression plasmid in the cell cycle-arrested MCF-12A cells induced c-Jun protein expression, and co-injection of dominant negative mutants of Ras, Rac, and Rho suppressed the transcriptional activity of c-Jun in the presence of BCAR3. Furthermore, stable expression of BCAR3 increased the proliferation of MCF-12A cells. The microinjection of inhibitory materials such as anti-BCAR3 antibody and siRNA BCAR3 inhibited EGF-induced c-Jun expression but did not affect IGF-1 induced upregulation of c-Jun. Taken together, we propose that BCAR3 plays a crucial role in c-Jun protein expression and cell proliferation and that small GTPases (e.g., Ras, Rac, and Rho) are required for the BCAR3-mediated activation of c-Jun expression.

• Korean Journal of Microbiology
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• v.46 no.2
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• pp.140-147
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• 2010

Japanese encephalitis virus (JEV), a member of the mosquito-borne flaviviruses, causes epidemics of viral encephalitis in the Southeastern Asia. JEV is a small enveloped virus with a positive-sense RNA genome; the infectious virion consists of three structural proteins, namely capsid, membrane (M; a mature form of its prM precursor), and envelope proteins. Here, we investigated a role of the charged residues found at the N-terminus of the JEV M protein in virus production. Using an infectious JEV cDNA, we generated two mutant cDNAs, Mm1 and Mm2, by charged-to-alanine substitution for $E^9$ and $K^{15}K^{16}E^{17}$ residues of the M protein, respectively. By transfection of wild-type or each of the two mutant RNAs transcribed from the corresponding cDNAs, we found that Mm2, but not Mm1, had a ~3-log decrease in virus production, even though a comparable amount of all three structural proteins were produced in transfected cells. Interestingly, the prM protein expressed in Mm2 RNA-transfected cells was not recognized by the polyclonal antiserum raised against the N-terminal 44 amino acids of the wild type M protein, but reacted to the antiserum raised against the corresponding region of the mutant Mm2. Our results indicate that three charged residues ($K^{15}K^{16}E^{17}$) in JEV M protein play a role in virus production. Two polyclonal antisera specifically recognizing the wild-type or Mm2 version of the M protein would provide a useful reagent for the functional study of this protein in the virus life cycle.

• Journal of Chest Surgery
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• v.41 no.1
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• pp.12-24
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• 2008

Background: It is currently thought that tissue valve degeneration is related to an animal's immune response, which is mainly due to cell surface ${\alpha}$-Gal epitopes. Cell surface ${\alpha}$-Gal epitopes are known to be degraded by the enzyme called green coffee bean ${\alpha}$-Galactosidase. It is also well known that ${\alpha}$-Gal epitopes are immunologically stained by Griffonia Simplicifolia isolectin type B4. We know that many commercially available tissue valves are made of aortic valves and pericardial tissue of pig. So, we investigated whether ${\alpha}$-Gal epitopes of the aortic valve and pericardial tissue of a pig can be removed by green coffee bean ${\alpha}$-Galactosidase, and we did so by comparing immunologic staining of the tissues before and after the enzyme treatment. Material and method: After treating fresh porcine aortic valve and pericardial tissue with green coffee bean ${\alpha}$-Galactosidase at concentrations of 0.5 unit/mL, 1.0 unit/mL, 2.0 unit/mL, respectively, under the condition of pH 6.5, temperature. $4^{\circ}C$ and 24 hours of incubation, each sample was stained with Griffonia Simplicifolia isolectin type B4 immunpfluorescent labeling. We then examined whether the ${\alpha}$-Gal epitopes were reduced or abolished in each consecutive. concentration of green coffee bean ${\alpha}$-Galactosidase by comparing the degree of the Griffonia Simplicifolia isolectin B4 staining in each sample. Result: In the pig aortic valve tissue, a 1.0 unit/mL concentration of green coffee bean ${\alpha}$-Galactosidase at pH 6.5, $4^{\circ}C$ and reaction for 24 hours was enough for complete removal of ${\alpha}$-Gal epitopes from the cell sur face on the immunostaining with Griffonia Simplicifolia isolectin B4. On the other hand, more ${\alpha}$-Gal epitopes were present in the pig pericardial tissue on Griffonia Simplicifolia isolectin B4 staining before the enzyme treatment, and 1.0 unit/mL of galactosidase was not sufficient for complete removal of ${\alpha}$-Gal from the tissue. 2.0 units/mL of green coffee bean ${\alpha}$-Galactosidase was needed to completely remove the ${\alpha}$-Gal epitopes from the pericardial tissue on immunostaining. Conclusion: The ${\alpha}$-Gal epitopes of the pig's aortic valve and pericardial tissue were successfully stained with Griffonia Simplicifolia isolectin B4. We could remove nearly all the ${\alpha}$-Gal epitopes using green coffee bean ${\alpha}$-Galactosidase at the concentration of 1.0 unit/mL in the aortic valve. Of pig, and 2.0 unit/mL was need to nearly completely remove all the ${\alpha}$-Gal epitopes in the pericardial tissue of pig under the condition of pH 6.5, $4^{\circ}C$ and 24 hours of reaction time. In the near future, removal of ${\alpha}$-Gal epitapes in the pig's aortic valve and pericardial tissue will become a powerful tool for the improvement of the tissue valve durability. It needs to be determined if ${\alpha}$-galactosidase treated pig tissue is immune to human anti-Gal antibody or anit-Gal mooclonal antibodies.