• Journal of Microbiology and Biotechnology
• /
• v.18 no.7
• /
• pp.1326-1334
• /
• 2008

The prime-boost vaccination with DNA vaccine and recombinant viral vector has emerged as an effective prophylactic strategy to control infectious diseases. Here, we compared the protective immunities induced by multiple alternating immunizations with DNA vaccine (pCIgB) and replication-incompetent adenovirus (Ad-gB) expressing glycoprotein gB of pseudorabies virus (PrV). The platform of pCIgB-prime and Ad-gB-boost induced the most effective immune responses and provided protection against virulent PrV infection. However, priming with pCIgB prior to vaccinating animals by the DNA vaccine-prime and Ad-boost protocol provided neither effective immune responses nor protection against PrV. Similarly, boosting with Ad-gB following immunization with DNA vaccine-prime and Ad-boost showed no significant responses. Moreover, whereas the administration of Ad-gB for primary immunization induced Th2-type-biased immunity, priming with pCIgB induced Th1-type-biased immunity, as judged by the production of PrV-specific IgG isotypes and cytokine IFN-$\gamma$. These results indicate that the order and injection frequency of vaccine vehicles used for heterologous prime-boost vaccination affect the magnitude and nature of the immunity. Therefore, our demonstration implies that the prime-boost protocol should be carefully considered and selected to induce the desired immune responses.

• Parasites, Hosts and Diseases
• /
• v.25 no.2
• /
• pp.141-148
• /
• 1987

Circulating antigens in rats experimentally infected with Paragonimus westermani were examined by ELISA. From a total of 22 albino rats, each fed with 25 metacercariae, blood samples were collected until 12 weeks after infection. The specific antibodies against P. westermani in the serum of an infected cat were purified by ammonium sulfate precipitation, DEAE anion-exchange chromatography and affinity chromatography serially. So-called double antibody sandwich ELISA method was used for the detection of circulating antigens. The results were as follows: Mean value of O.D. in control sera was O. 04 (S.D.=0. 04). After infection, mean O.D.(S.D.) values were changed serially: 0.03(0.01) at 0.5 week(3 days), 0.55(0.50) at 1 week, 0.69(0.45) at 1.5 week, O.20 (0.19) at 2 weeks and O.13(0.10) at 2.5 weeks of infection. They returned, thereafter, to the level before infection. When O. 16 (mean+3 S.D.) were considered as cut-off value, those higher than O. 16 were observed only in the sera collected between 1 and 2.5 weeks after infection. Average 8. 4 immature worms (2.2 from the lungs and pleural cavities; 6.2 from muscles) were recovered in a rat at 12 weeks after infection. The fact that circulating antigens were not detected after 3 weeks of infection was considered to the caused by the formation of antigen-antibady complexs.

• Journal of fish pathology
• /
• v.23 no.2
• /
• pp.211-219
• /
• 2010

Specimens were collected from the coastal region near industrial complex of Ulsan-Onsan, Sihwa-Ansan and Yeosu-Gwangyang in 2008 and 2009. The total number of individuals used in analysis was 1,289 of Acanthogobius flavimanus, Chelon haematocheilus, Hemibarbus labeo, Leiognathus nuchalis, Mugil cephalus and Synechogobius hasta. The sex ratio in the total individual was 1:0.73 (female:male). Specific sex ratio of fishes in the areas, namely the Ulsan-Onsan, Sihwa-Ansan and Yeosu-Gwangyang were 1:0.79, 1:0.81, and 1:0.25, respectively. Especially, female in Yeosu-Gwangyang was higher than male. The intersexuality in the total individual was 11.7%. Intersexuality of fishes in the areas were 4.98, 14.39 and 25.0% in the Ulsan-Onsan, Sihwa-Ansan and Yeosu-Gwangyang, respectively. It was indicated female higher than male in Ulsan-Onsan and male higher than female in Sihwa-Ansan and Yeosu-Gwangyang.

• Parasites, Hosts and Diseases
• /
• v.26 no.2
• /
• pp.73-86
• /
• 1988

The sensitivity and specificity of crude and affinity-purified antigens of Clcnorchis sinensis obtained from the infected rabbits were studied. Stage-specific antigenic proteins from the eggs, metacercariae and adult worms were characterized by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and enzyme-linked immunosorbent astray (ELISA). The results were as follows: 1. The antibody.binding antigen (ABA) purified from whole worm crude antigen (IVWA) by CNBr-activated Sepharose 4B affinity chromatography made :l specific bands against rabbit antisera on Ouchterlony gel diffusion plate, while WWA made 7 bands. Major WWA protein bands by SDS-PAGE were found at 16, 300~18, 500 and 28, 000~29, 000 daltons, while major ABA protein bands were at 18, 000~21, 000 and 29, 000~31, 000 daltons. The reactivity of ABA with rabbit anti-sera in ELISA was remarkably less sensitive than that of WWA. 2. Molecular weights of egg antigen (EGA), metacercarial antigen (MEA) and adult worm antigen (WWA) of C. sinensis ranged from 15, 000-200, 000 daltons, 15, 000-100, 000 daltons and 11, 000~80, 000 daltons, respectively. Major WWA proteins consisted mainly of polypeptide bands of low molecular weight, less than 31, 000 daltons, while those of EGA and MEA consisted of higher molecular T.eights than 30, 000 daltons. 3. The ELISA reactivities of WWA to rabbit anti.sera were remarkably greater than those of MEA. EGA showed negative reaction throughout the experiments. WWA showed higher optical density (O.D.) than 1.0, when reacted with rabbit anti-sera obtained at 4~6 weeks after the infection. In the rabbit anti-sera later than 12 weeks after the infection, the O.D. reacting witll WWA showed a plateau without variation. MEA shoT.ed relatively low O.D. values (<0.6), when reacted with anti-sera from lightly in(ected groups throughout the experiments, althougll there were some wealth positive cases (O.D.>0.6) ill heavily infected groups. MEA reacted with rabbit anti-sera showed negative results on Ouchterlony gel diffusion plates. Summarizing the above results, it is suggested that the whole worm antigen prepared from the adult worms of C. sinensis is most highly antigenic. However, this antigen might reveal cross reactions with other trematodes such as Paragonimus westermani, therefore, purification of antigenic proteins from the crude antigen is essential 18 increase the sensitivity and specificity for the immuncdiagnosis of clonorchiasis.

• Clinical and Experimental Pediatrics
• /
• v.50 no.6
• /
• pp.556-564
• /
• 2007

Purpose : We hypothesized that the persisting bronchial hyperresponsiveness (BHR) of adolescents with asthma remission may be controlled mainly by genetic factors, and the BHR of symptomatic asthma by airway inflammation. ${\beta}_2$-adrenoceptor gene is considered to be a candidate gene in the development of BHR. Thus, ${\beta}_2$-adrenoceptor gene polymorphism may be associated with the BHR of adolescents with asthma remission, but not with the BHR of symptomatic asthma. To evaluate this hypothesis, ${\beta}_2$-adrenoceptor gene polymorphism at 2 sites (Arg16-Gly, Gln27-Glu) were examined. Methods : Two hundred two adolescents with BHR ($PC_{20}<18\;mg/mL$) and long term remission (neither asthma-related symptoms nor medication during the previous 2 years) of their asthma (remission group), 182 adolescents with symptomatic asthma (symptomatic group), and 200 healthy adolescents (control group) were studied. Asthma phenotypes were determined using methacholine bronchial provocation test and skin prick test. Genotypes of ${\beta}_2$-adrenoceptor polymorphism were evaluated by PCR-based methods. Results : Gly/Gly allele and Gly16-Gln27 haplotype were more prevalent in the remission group than in the control group (P=0.01, P=0.02), although there was no difference between the symptomatic group and the control group. In the remission group, there was significant difference in geometric mean of $PC_{20}$ among the 3 groups subdivided by the number of Gly16-Gln27 haplotype, showing that the Gly16-Gln27 haplotype was positively associated with BHR. However, no association was found between Gly16-Gln27 haplotype and BHR in the symptomatic group. Conclusion : This study demonstrates that ${\beta}_2$-adrenoceptor polymorphism at amino acid 16 and 27 was associated with BHR persisting in adolescents with asthma remission.

• Korean Journal of Ecology and Environment
• /
• v.44 no.3
• /
• pp.272-279
• /
• 2011

Copper cyanide is a chemical produced in large quantities with 2,500 tonnes being produced in 2006. It is mainly used for electroplating copper, particularly alkali-Cu plate and brass plating. The purpose of this study is to reassess the physicochemical properties and environmental fate of copper cyanide based on reliable data and and to conduct an ecotoxicity test according to the OECD test guidelines as an initial environmental risk assessment (need to state where this was done). Metal containing inorganic substances are not subject to degradation, biodegradation or hydrolysis. Aquatic toxicity tests of copper cyanide were conducted according to OECD test guideline 201, 202 and 203 for green algae, daphnia, and fish, respectively. The following acute toxicity test results were obtained for aquatic species: 0.089 mg $L^{-1}$ (Algae, 72 Hr-$EC_{50}$); 0.21 mg $L^{-1}$ (flea, 48 Hr-$LC_{50}$); 0.62 mg $L^{-1}$ (Fish, 96 Hr-$ErC_{50}$). The chemical possesses properties indicating a hazard for the aquatic environment (acute toxicity in fish, daphnia and algae below 1.0 mg $L^{-1}$). As a result of this study, copper cyanide has become a candidate for detailed risk assessment. Countries that produce this chemical in significant quantities are recommended to perform specific assessments.

• Food Science of Animal Resources
• /
• v.29 no.4
• /
• pp.445-456
• /
• 2009

Colostrum samples were collected from 36 dairy farms in Gyeonggi-do and one dairy farm in the National Institute of Animal Science (NIAS) for testing. Colostrum samples were analyzed for phisycochemicals (specific gravity, pH, titratable acidity), general components (fat, protein, lactose, total solid, solid non-fat (SNF)), fatty acids, amino acids, minerals, microflora, somatic cells, and Ig (Immunoglobulin). The first colostrum revealed the following data: fat contents were $6.16{\pm}2.39%$, proteins were $14.78{\pm}4.30%$, lactose $2.57{\pm}0.77%$, total solid $24.28{\pm}4.36%$, and SNF $18.12{\pm}4.08%$, whereas the 2nd (or $12^{th}$) colostrum revealed $5.56{\pm}1.76%$ fat, $3.46{\pm}0.41%$ proteins, $4.19{\pm}0.43%$ lactose, $13.90{\pm}1.76%$ total solid, and $8.34{\pm}0.81%$ SNF. Also, the first colostrum revealed the contents of major amino acids as 0.89% aspartic acid, 0.71% threonine, 0.86% serine, 1.75% glutamic acid, 0.64% valine, 0.95% leucine, 0.83% lysine, and 0.95% proline, and those in the 10th colostrum were 0.25% aspartic acid, 0.15% threonine, 0.19% serine, 0.59% glutamic acid, 0.19% valine, 0.35% leucine, 0.31% lysine, and 0.34% proine. Major amino acid contents rapidly decreased as milking times increased. In the first colostrum, the following mineral contents were observed: there were 2,168 ppm in Ca, 1,959 ppm in P, 914 ppm in K, 761 ppm in Na, 287 ppm in Mg, 1.7 ppm in Fe, 14.3 ppm in Zn, and 1.0 ppm in Cu; while in the 10th colostrum, the following ppm contents were 1,389 in Ca, 1,323 in P, 838 in K, 427 in Na, 131 in Mg, 1.0 in Fe, 4.7 in Zn, and 1.3 in Cu. The mineral contents in a colostrum rapidly decreased as milking times increased.

• Journal of Life Science
• /
• v.16 no.5
• /
• pp.780-787
• /
• 2006

Asthma is a chronic inflammatory disorder of the airways, which characterized by bronchial hyperresponsiveness, reversible airflow limitation and respiratory symptoms. Internationally, the prevalence of asthma has been increased over last 3 decades. Recently, several studies of asthma have been reported with gradually increasing importance. To tesify the hypothesis that interleukin (IL)-4 and IL-10 may be an important determinant of the severity of airway inflammation, their expression was studied in mouse model of asthma. BALB/c mouse, IL-4 Knockout (KO) mouse and IL-10 KO mouse were sensitized with intraperitoneal injection of ovalbumin adsorbed to aluminum potassium sulfate, followed by challenges with intranasal ovalbumin on 3 consecutive days. The severity of pulmonary inflammation was assessed by eosinophilia in BAL fluid, number of total BAL cells, histopathological changes in lung tissues, and immunohistochemical staining against IL-4 and IL-10. In BAL fluid, the number of total cells was significantly increased in asthma induced mouse compare to the control. In asthma induced mouse, eosinophil was increased to 56% and neutrophil was 0.2%. In H &E stains, eosinophilic infiltration and epithelium hyperplasia were clearly noticed in asthma induced mouse. In immunohistochemical staining for IL-4 and IL-10, there was no positive reaction in control group. However, very strong reactions were appeared in asthma induced group. In this research, IL-4 and IL-10, which seem to play a central role in allergic asthma, KO mouse was utilized to test the causative relationship between airway inflammation and role of specific cytokine. Asthma induced IL-4 and IL-10 KO mice showed much decreased inflammatory reactions in the number of total BAL cells, in eosinophilic infiltration, and in immunohistochemical stains against diverse inflammatory proteins. These results suggest that IL-4 and IL-10 increase the asthmatic reactions in vivo mice model.

• The Journal of the Korean Society for Microbiology
• /
• v.21 no.1
• /
• pp.151-161
• /
• 1986

In order to develop sensitive and sepcific assay methods for E. coli heat labile enterotoxin(LT) hybridoma cell lines secreting LT specific monoclonal antibody were obtained. LT was purified from cell lysate of E. coli O15H11. The steps included disruption of bacteria by French pressure, DEAE Sephacel ion exchange chromatography, Sephadex G200 gel filtration, and second DEAE Sephacel ion exchange chromatography, successively. Spleen cells from Balb/c mice immunized with the purified LT and $HGPRT^{(-)}$ plasmacytomas, $P3{\times}63Ag8.V653$ were mixed and fused by 50% (w/v) PEG. Hybrid cells were grown in 308 wells out of 360 wells, and 13 wells out of them secreted antibodies reacting to LT. Among these hybridoma cell 1G8-1D1 cell line was selected since it had produced high-titered monoclonal antibody continuously. By using culture supernatant and ascites from 1G8-1D1 cells the monoclonal antibody was characterized, and an assay system for detecting enterotoxigenic E. coli was established by double sandwich enzyme-linked immunosorbent assay (ELISA). The following results were obtained. 1. Antibody titers of culture supernatant and ascites from 1G8-1D1 hybridoma cells were 512, and 102, 400, respectively by GM1-ELISA and its immunoglobulin class was IgM. 2. The maximum absorption ratio of 1G8-1D1 cell culture supernatant to LT was 90% at $300\;{\mu}g/ml$ of LT concentration. LT concentration shown at 50% absorption ratio was $103.45{\mu}g$ and the absorption ratio was decreased with tile reduction of LT concentration. This result suggests that monoclonal antibody from 1G8-1D1 hybridoma cell bound with LT specifically. 3. The reactivities of 1G8-1D1 cell culture supernatant to LT and V. cholerae enterotoxin(CT) were 0.886 and 0.142(O.D. at 492nm) measured by the GM1-ELISA, indicating 1G8-1D1 monoclonal antibody reacted specifically with LT but not with CT. 4. The addition of 0.1ml of ascites to 0.6mg and 0.12mg of LT decreased the vascular permeability factor to 41% and 44% respectively, but it did not completely neutralize LT. 5. By double sandwich ELISA using monoclonal antibody, as little as 75ng of the purified LT per ml could be detected. 6. The results by assay of detecting LT in culture supernatants of 14 wild strains E. coli isolated from diarrhea patients by the double sandwich ELISA were almost the same level as those by reverse passive latex agglutination.