• Korean Journal of Veterinary Research
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• v.32 no.3
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• pp.451-456
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• 1992

Seventeen antiserum identified by stormont method were used for investigation of appearance type of red blood cell in Cheju horse. Phenotype $A_1$, A', Z, $ZZ_2$, C, K, $P_1$, Q, R, S, $U_2$, X and $N_1$ were classified by Hemolytic reaction and agglutination in $E_2$, H, J and $T_1$, Hokkaido horse and Thoroughbred horse were compared with the type of red blood cell. The results obtained are as follows ; 1. According to the appearance frequency of red blood cell the phenotypes could be classified by 6 groups among horses tested; I group : C, $A_1$, $ZZ_2$, $E_2$, S, Q, II : $P_1$, $U_2$, X, III : A', R, IV : $T_1$, K, V : H, J, VI, $N_1$, Z. 2. In group I the type of C, $A_1$ and $ZZ_2$ showed the Similar frequency among horses but the type of $E_2$, S and Q was lower in Cheju horse than Hokkaido and Thoroughbred horse. 3. In group II there was no differences in the type of $P_1$ and among three species, but Thoroughbred horse was lowest as 11.2 percent than Cheju and Hokkaido horse. 4. The type of $U_2$ and X were highest in Cheju and Hokkaido horse but lowest in Thoroughbred horse. 5. The type of A' and R observed higher appearance frequency in Hoklcaido horse but in the Thoroughbred and Cheju horse appearance ratio were Strikingly lowered. 6. The type of $T_1$ and K was shown higher appearance frequency in Cheju and Thoroughbred horse similarily but lowered in Hokkaido horse. 7. In group V the type of H and J showed similarily as 30 to 40 percent in Cheju and Thoroughbred but no observation in Hokkaido horse. 8. The type of $N_1$ and Z which are group VI was great difference between Cheju and Hokkaido horse but higher in Hokkaido than Cheju horse while there was no observation in thoroughbred horse.

• Journal of fish pathology
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• v.14 no.2
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• pp.97-102
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• 2001

Olive flounder, Paralichthys olivaceus known as an one of the major aquacultured species in Korea were exposed to cadmium(Cd) with different protocols and analyzed the effects of exposure on the immune response. Antibody levels in sera of the group exposed to Cd(20ppb) by immersion method from 2 weeks before immuniztion with formalinised Edwardsiella tarda(E. tarda) KFE entigen to the end of experiment reached to peak level faster than that of the non-exposed group. After this peaking time the levels decreased much at a faster rate compared to the non-exposed group. This tendency was also appeared in the numbers of specific antibody secreting cells(SASC) analyzed with the enzyme-linked immunospot (ELISPOT)-assay technique in the splenocytes of the experimental groups exposed to Cd with different ways. Interestingly, the group exposed to Cd for 2 weeks before immunization also showed increased numbers of SASC unlikely the antibody production and suggested a more critical influence of cadmium exposure in early stage of immune reaction. Artificial infection with live E. tarda KFE induces 100% mortality in the flounder exposed to cadmium throughout the experimental period from two weeks before the immunization. It may imply that some other factors related to specific immunity are involving in the defence system of flounder exposed to Cd. Taked together. Cd exposure may induce temporaily stimulatory or indhibitory effects on the immune reaction, but suppress the physiological systems for the resistant against the infective agents with other toxic effects.

• Korean Journal of Veterinary Research
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• v.48 no.1
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• pp.53-60
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• 2008

We have investigated efficiency of a recombinant subunit Pasteurella multocida toxin (PMT) that was mixed with a vaccine consisted of inactivated whole cells of Bordetella bronchiseptica, P. multocida (types A and D). For verification of the efficacy of the vaccine, all experimental pigs (suckling piglets, sow and gilts) in the three farms were vaccinated. Antibody titers against B. bronchiseptica and P. multocida type A of the vaccinated pigs by microplate agglutination were significantly higher than those of the control pigs (p < 0.05). Similar patterns were observed in the analysis of anti- PMT neutralizing antibody by serum neutralizing method using Vero cell (p < 0.05). Anti- P. multocida type D antibody titer of the vaccinated sows and gilts by ELISA showed significant differences with those of the non-vaccinated pigs (p < 0.05). Although antibody titers increased, it was unable to find out the difference in the clinical signs between the vaccinated and non-vaccinated pigs. However, the increase in body weight of the vaccinated piglets was observed in comparison with the non-vaccinated piglets on a farm. At slaughtering of the pigs, pathological lesions in the turbinate bones of the vaccinated pigs were significantly lower than those of the non-vaccinated pigs (p < 0.001). These results suggested that efficacy of the vaccine in pigs demonstrated to protect against atrophic rhinitis in Korea.

• Journal of Microbiology and Biotechnology
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• v.19 no.10
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• pp.1238-1243
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• 2009

The susceptibilities of major enterohemorrhagic Escherichia coli (EHEC) strains to antimicrobial agents and the cytotoxicity of these agents were examined using a total of 38 strains of E. coli O26, O111, and O157, which are the major serogroups of EHEC. Among the 38 strains, 35, 36, and 36 were susceptible to amikacin, imipenem, and norfloxacin, respectively. These antimicrobial agents were further examined to determine their cytotoxicity on Vero cells as well as their effect on the release of Shiga toxins along with trimethoprim/sulfamethoxazole. Each of the E. coli O26, O111, and O157 strains containing both the stx1 and stx2 genes were grown in the absence or presence of these agents at 1/4 minimal inhibitory concentration for 6 h, 12 h, and 18 h. At the concentrations used in this study, none of the agents significantly altered cell count compared with the control group. The level of cytotoxicity in the imipenem group was lower at 12 hand 18 h than their respective controls. In contrast, the level of cytotoxicity in cultures treated with trimethoprim/sulfamethoxazole, norfloxacin, and amikacin was increased. The strains were also examined for the release of Shiga toxins 1 and 2 following treatment with the agents, which were measured by the reversed passive latex agglutination (RPLA) method. The RPLA assay showed a suppression of release of Shiga toxin 2 in the strain cultures containing imipenem. These results indicate that imipenem may be a safe and effective agent for inhibition of these bacteria, which has clinical implications for the treatment of EHEC infections.

• Environmental Analysis Health and Toxicology
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• v.21 no.2 s.53
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• pp.165-172
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• 2006

The backs with a hair cut of 6-week-old healthy ICR male mice were once exposed to a dose of $400mJ/cm^2$ UVB. An acute dermal inflammation was observed, and the inflamed skins were almost completely cured after 6 days of the exposure. At 24 hours after exposure, the epidermal keratinocytes showed a cell-membrane damage with the destruction of intercellular junctions, agglutination of tonofilaments within the cytoplasm and nucleus damage. The activity of XO showed a significant increase (p<0.05) in up to 144 hours. The activities of CAT and SOD showed a significant decrease (p<0.05) in up to 96 hours, but they were not significantly different from the normal value at 144 hours. The GST activity was significantly decreased (p<0.01) in up to 96 hours, not so at 24 hours. However, that was not significantly different from the normal value at 144 hours. There was a significant decrease (p<0.01) in the contents of TBARS at 48 and 96 hours, without any significant difference at 144 hours. While the content of GSH was significantly lower (p<0.05) at 24 hours, that was not significantly different thereafter up to 144 hours from the normal value. Therefore, it is assumed that skin damage with a dose of $400mJ/cm^2$ UVB irradiation might be caused by the oxidative stress which was resulted from the unbalance of oxygen fret radical generating and scavenging enzymes.

• Korean Journal of Microbiology
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• v.37 no.4
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• pp.289-293
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• 2001

Incidence of infectious viruses is ensuing throughout the world and threatening the health of children as well as adults. The outbreaks of viral diseases of alimentary tract in Pusan from 1998 to 2000 were detected. Viruses were isolated from stool specimens, cerebrospinal fluid and throat swabs from suspicious patients and confirmed by cell culture, latex agglutination test, indirect immunofluorescent test and electron microscopic observation. The average isolation rate was 12.5% from the suspected specimens. From this work, 2 cases of enteric adenoviruses, 23 cases of echovirus, 31 cases of coxsackivirus 36 cases of rotavirus, 45 cases of SRSV, and 7 cases of poliovirus were detected. The major serotypes of coxsackievirus were B2, B3, B4, B6 and echovirus of serotypes 6, 9, 11, 25, and 30 were examined. Two cases of enteric adenovirus type 41 were also confirmed. The incidence of SRSV was mostly concentrated between December through following March, April through October with echovirus and coxsackievirus, and January through April with rotavirus, respectively. Electron micrograph of negative-stained viruses showed typical appearance with 30-80 nm in diameter.

• Journal of Life Science
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• v.18 no.2
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• pp.241-248
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• 2008

This study has been carried out to secretion antibodies for the purpose of preventing the infection of Helicobacter pylori and using them as a supplement for treatment. This experiments have been separated antigens from H. pylori and observed into antibody production and the agglutination of H. pylori for the separated antigens. As major antigenic proteins separated from H. pylori, the following could be verified: 12 kinds of band for whole cell (WC), seven kinds of band for outer membrane protein (OMP), three kinds of band for crude urease, and one kind of band for lipopolysaccharide (LPS). The IgG anti-H. pylori antibody of separated antigens showed $77.9{\pm}6.4{\mu}g/ml$ for we (L), $84.9{\pm}6.4{\mu}g/ml$ for OMP, and $123.8{\pm}2.9{\mu}g/ml$ for crude urease, at the same antigen concentration of $20{\mu}g/100ull$, which showed the most at the crude urease. And it turned out that the IgA antibodies were generated with $2.5{\pm}0.32{\mu}g/ml$ for WC (L), $2.0{\pm}0.43{\mu}g/ml$ for OMP, and $1.3{\pm}0.25{\mu}g/ml$ for crude urease, which demonstrated the most for WC (L) antigens. As a result of verifying the immunogenecity of antigenic protein through the Western blotting, major antigenic substances could be confirmed as follows: 10 kinds for WC, six kinds for OMP and three kinds for crude urease. The agglutination values on the H. pylori of the antibody were $2^5,\;2^5,\;2^6\;and\;2^7$ at the antigen serums of anti-WC (H), anti-WC (L), anti-OMP and anti-crude urease, respectively, which indicated the highest for the antigen serum of anti-crude urease. The urease activation-inhibiting absorbance of antigen serum created by each antigen was $0.14{\pm}0.01$ for WC (H), $0.16{\pm}0.01$ for WC (L), $0.18{\pm}0.03$ for OMP, and $0.18{\pm}0.04$ for urease, demonstrating a significant inhibiting effect, compared with $0.26{\pm}0.02$ of the control group.

• The Journal of the Korean Society for Microbiology
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• v.18 no.1
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• pp.73-85
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• 1983

The advantages of the enzyme-linked immunosorbent assay(ELISA) are its sensitivity and its simplicity in detecting IgM and IgG antibodies. For applying the ELISA to the diagnosis of typhoid fever, first of all, experiments were performed to determine which concentration of killed whole cell antigens and lipopolysaccharide(LPS) antigens of S. typhi(0.901 w) were optimally coated to the wells of the polystyrene and polyvinylchloride microplate, using the hyperimmune sera from rabbits against S. typhi. By using both kinds of antigens of S. typhi adsorbed to the ELISA microplate, the changing patterns of IgG and IgM antibodies in the sera from rabbits responding to the killed whole cell antigens of S. typhi(0901 w) during the prolonged immunization were serially traced by the ELISA. At the same time, the level of antibodies against S. typhi in sera fron patients with typhoid fever and from normal healthy persons were measured by the ELISA employing the killed whole cell antigens and LPS antigens as the coating antigens. The results obtained were summerized as follow: 1. The optimal concentration of the killed whole cell antigens, which were more easily adsorbed to the polystyrene plate than the polyvinylchloride plate, was $10^8cells/ml$ of carbonate buffer(pH. 9.6) on the wells of the polystyrene plate when treated at $37^{\circ}C$ for 4 hours. On the other hand, the optimal concentration of lipopolysaccharide antigens, which were adsorbed only to the polyvinylchloride plate, was $100{\mu}g/ml$ of carbonate buffer(pH. 9.6) on the wells of the polyvinylchloride plate when treated at $37^{\circ}C$ for 4 hours. 2. IgM antibody response were dominating in rabbits responding to the killed whole cell antigens of S. typhi(0.901 w), and were more specific to the LPS antigens than to the killed whole cell antigens in the ELISA. Good correlations were made between the IgM titers by the ELISA and the aggglutinating titers of sera from the immunized rabbits. 3. Both IgG and IgM agglutination titers by the ELISA in sera from most of patients with typhoid fever were above 1:320 but those in sera from most of normal, healthy persons were below 1:80. 4. There were close correlations between the antibody titers by the ELISA and the agglutinating titers to the killed whole cell antigens in the tested human sera, IgM titers being more correlated with the agglutinating titers than IgG titers. But a little correlations were made between the antibody titers by the ELISA and the agglutinating titers to the LPS antigens. 5. IgM titers in the tested human sera were similar to IgG titers detected by the ELISA employing the killd whole cells antigens and the LPS antigens. 6. Good correlations were made between the antibody titers demonstrated by the ELISA performed on the killed whole cell antigens and the LPS antigens as the different, coating antigens on the ELISA microplates.

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

• Journal of the korean academy of Pediatric Dentistry
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• v.30 no.4
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• pp.554-562
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• 2003

Xylitol has been used as sugar substitute to prevent dental caries. It is not fermented by most dental plaque bacteria and interferes with the growth of mutans streptococci. Therefore the production of acidic metabolites and the growth of mutans streptococci are inhibited. S. mutans strains which are inhibited to grow under the presence of xylitol are referred as xylitol-sensitive ($X^S$) strains. However, experimental and clinical studies have shown that there were mutated groups of S. mutans strains that are not affected by xylitol. They are referred as xylitol-resistant($X^R$) strains. The aim of the present study was to investigate that emergence of $X^R$ strain would effect on the anticariogenecity of xylitol by comparing the growth rate, the extracellular pH, hydroxyapatite adhesion and the agglutination of the $X^R/X^S$ strains. Overall we came out with following results : 1. No difference in the growth rate and the extracellular pH was found between the $X^S$ strain and the $X^R$ strain. 2. No difference in adhesion to hydroxyapatite surface was found between the $X^R$ strain and the $X^S$ strain (p>0.05) and adhesion of the $X^S$ strain was greater than that of $X^R$ strain in the sucrose-dependent adhesion to hydroxyapatite (p<0.05). 3. The $X^R$ strain was agglutinated in the lower concentration of saliva than that of $X^S$ strains.