• Molecules and Cells
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• v.22 no.1
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• pp.104-112
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• 2006

We previously reported that DA-9601, ethanol herbal extract of Artemisia asiatica, inhibited histamine and leukotriene releases in guinea pig lung mast cells activated with specific antigen/antibody reaction. This study aimed to evaluate the inhibitory effect of DA-9601 on the OVA-induced airway inflammation in allergic asthma mouse model. BALB/c mice were sensitized and challenged with OVA. DA-9601 was administered orally 1 h before every local OVA-challenge. OVA-specific serum IgE was measured by ELISA, recruitment of inflammatory cells in BAL fluids and lung tissues by Diff-Quik and H&E staining, respectively, the expressions of CD40, CD40L and VCAM-1 by immunohistochemistry, goblet cell hyperplasia by PAS staining, activities of MMPs by gelatin zymography, expressions of mRNA and proteins of cytokines by RT-PCR and ELISA, activities of MAP kinases by western blot, and activity of NF-${\kappa}B$ by EMSA. DA-9601 reduced IgE level, recruitment of inflammatory cells into the BAL fluid and lung tissues, expressions of CD40, CD40L and VCAM-1 molecules, goblet cell hyperplasia, MMPs activity, expressions of mRNA and productions of various cytokines, activities of MAP kinases and NK-${\kappa}B$ increased from OVA-challenged mice. These data suggest that DA-9601 may be developed as a clinical therapeutic agent in allergic diseases due to suppressing the airway allergic inflammation via regulation of various cellular molecules expressed by MAP kinases/NF-${\kappa}B$ pathway.

• Korean Journal of Veterinary Service
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• v.31 no.1
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• pp.57-70
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• 2008

The purpose of this study was to investigate the protective effects of the modified live vaccines against canine Bordetella bronchiseptica (Bb) and canine parainfluenza virus (CPIV) in puppies by nasal administration. A total of 24 puppies were classified as 3 groups consisting of 8, and each one was divided into two subgroups; vaccinated (n=4) and unvaccinated (n=4). Group I, group II and group III were challenged with Bb, CPIV, and Bb+CPIV, respectively. In group I vaccinated puppies (n=4) were experimentally challenged with Bb 2 weeks after vaccination and unvaccinated puppies (n=4) were experimentally challenged with Bb alone. The same methods of the above were applied for group II and group III. We carried out several studies including serum tests, isolation, and histopathological examination. The vaccinated puppies showed higher antibody titers than unvaccinated puppies and the titer sustained during the experimental period. In the isolation test, recovery period was shorter in the vaccinated subgroup than in the other. In clinical signs, the unvaccinated puppies showed the typical signs of tracheobronchitis (coughing, nasal and occular discharge), but another subgroup showed delayed incidence and mild clinical signs. The typical gross lesions and histopathological findings were observed in the unvaccinated puppies. In immunohistochemical findings, the vaccinated puppies showed little intensive in reaction for CPIV antigen than the other. It could be concluded that intranasal vaccination of modified live Bb and CPIV vaccine to puppies is effective to prevent canine infectious tracheobronchitis.

• Journal of Sensor Science and Technology
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• v.6 no.5
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• pp.356-361
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• 1997

Fiber-optic evanescent wave sensor was designed and fabricated to detect mouse immunoglobulin G(IgG) with decladed optical fiber on which anti-mouse IgG was immobilized. A sensitivity obtained by any direct or competitive method was lower than $1\;{\mu}g/m{\ell}$. Anti-mouse IgG was immobilized on 93.9% of core surface of optical fiber by simple adsorption method. The effect of postcoating using bovine serum albumin to remove non-specific binding was not observed. As the ratio of fluorescein to mouse IgG increased, the fluorescence signal increased, but that increase showed no linear relationship. Our fiber-optic sensor system could be used as immunosensor by measuring evanescent fluorescence in antigen-antibody reaction with good sensitivity below $1{\mu}g/m{\ell}$ level.

• Parasites, Hosts and Diseases
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• v.34 no.4
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• pp.247-254
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• 1996

Differentiation of invasive strains of Entamoebn histolytica according to their pathogenicity has been a topic of long debate, but now the pathogenic species only is regarded as E. histolytica while the non-pathogenic species is E. dispar. The present study applied immunoblot to differentiale infections of the two species among microscopically- detected cyst-passers in Korea. The crude extract of 5. histolyticn separated in 5-20% gradient gels, revealed many fractions of 94. 81. 71, 50. 44, 38.5. 37.5, 29, 19. and 18 kDa when the cysteine proteinase inhibitor. E64, was supplemented. The serum IgG antibody of 3 proven E. histolytirc cases reacted loth the antigenic fractions of 117. 110. 99.68,66,60.54.52, 46. and 45 kDa. Sera of PCR confirmed 3 cases of E. disper reacted only to the 117 kDa fraction or the E. histolytica crude extract which was regarded as non specific. To the antitigen of monoxenic E. dispar. sera or E. dispar and E. histolytica cases showed the same immunoblot reactions. The serum IgG antibody reacted with several antigenic fractions of both E. histolytica and E. dispar. but IgM and IgE antibodies showed no reaction to either antigen. Sera of 24 symptomless amebic cyst-passers were screened with the E. histolytica alltigen; two were found to be infected by E. histolytica and 22 were by E. dispar. The present findings suggest that in Korea most of asymptomatic cyst passers of E. histolytica are carriers of E. dispar. Immunoblot using E. histolytica antigen is a good technique for the differentiation of E. histolytica and E. dispar infections.

• Parasites, Hosts and Diseases
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• v.28 no.4
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• pp.207-212
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• 1990

Enzyme-linked immunosorbent assay(ELISA) of paragonimiasis iloktsuenensis rat sera was performed using crude antigens of Paragonimus iloktsuenensis(PIA), P. westermani (PWA) and Clonorchis sinensis(CSA). Three crude antigens(PIA, PWA, CSA) were prepared to saline homogenated supernatants of whole adult worms. Infected rat sera were obtained biweekly from the albino rats fed 50∼.80 metacercariae of P. iloktsuenensis through gastric catheter. Experimental groups were divided into 4 groups: GI(controls), GII, GIII and GIV according to 1∼7 worms as GII, 10∼19 worms as GIII and 22∼40 worms as GIV, respectively, In ELISA, the mean OD values of each group for the homologous antigen(PIA) were increased significantly compared to the control sera at the 4th week of infection. With the progress of duration of infection, the mean OD values of infected sera of GII & GIV continuously increased up to the 12th week(last week), but in GIII the mean OD value increased until the loth week. No significance was noted among the infection dose groups (GII, GIII and GIV), after the 6th week of infection. Also, the OD values of all infected rats did not show any Proportional relytionships to the number of worms recovered. In brief, the antibody productivity of individual rats were strongly different. The rat sera infected with p. iloktsuenensis cross-reacted with those infected with P. westermani or C. sinensis, as identified by OD values.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2001.11a
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• pp.102-102
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• 2001

Taurine (2-ethaneaminosulfonic acid) is one of the major intracellular ${\beta}$ -amino acids in mammals and is required for a number of biological processes including membrane stabilization, osmoregulation, antioxidation, detoxification, modulation of calcium flux and neurornodulation. The taurine transporter (TAUT) which contains 12 hydrophobic membrane-spanning domains has been cloned from dog kidney, rat brain, mouse brain, human thyroid, placenta and retina. In this study, The TAUT cDNA from the human intestinal epithelial cell, HT-29 was cloned and sequenced. Reverse-transcription polymerase chain reaction (RT-PCR) was performed to amplify partial cDNA encoding human intestinal TAUT. The coding region of the PCR product was 732 bp long. The primers were designed to encode highly conserved amino acid sequences near the transmembrane domains III (IPYFIFLF) and Ⅵ (KYKYNSYR) both in human and mouse. The TAUT cDNA amplified was ligated into the pGEX 4T-1 expression vector. The resulting sequence of human intestinal TAUT cDNA (Accession number of NCBI Genebank is AF346763) was identical to the sequences of the TAUTs previously determined in the human placenta and retina except 3 base pairs from that of the reported human thyroid. TAUT specific antibodies were generated to use them as biological tools in the studies of the biological role of TAUT. Peptides of 149-162 amino acid residue (14 amino acids) of the TAUT were synthesized. The synthetic peptide used in this study was LFQSFQKELPWAHC. This region was chosen not only to avoid putative glycosylation sites but also to exclude regions of known homology with GABA transporters in the extracellular hydrophilic domains. The synthetic peptide, TAUT-1 was conjugated with carrier protein, kehole lympet hemocyanin (KLH) to use as an antigen. When used for immunization on a rabbit to produce polyclonal antiserum, the conjugates elicited high -titered specific anti-TAUT-1 antibodies, which reacted well with the ovalbumin (OVA) conjugated peptides in ELISA. The KLH-conjugated peptide was also used as immunizing antigen in BALB/c mice to produce TAUT specific monoclonal antibodies. From the culture supernatant of the hybridoma, the specificity of anti-TAUT-1 monoclonal antibodies was confirmed by ELISA. Further applications of more tools in TAUT expression analysis will be performed such as western blotting and flow cytometry.

• IMMUNE NETWORK
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• v.3 no.2
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• pp.126-135
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• 2003

Background: One of the important factors in the prognosis of chronic hepatitis B patient is the degree of replication of hepatitis B virus (HBV). It has been known that HBV DNA polymerase plays the essential role in the replication of HBV. HBV DNA polymerase is composed of four domains, TP (Terminal protein), spacer, RT (Reverse transcriptase) and RNaseH. Among these domains, tyrosine, the $65^{th}$ residue of TP is an important residue in protein-priming reaction that initiates reverse transcription. If monoclonal antibody that recognizes around tyrosine residue were selected, it could be applied to further study of HBV replication. Methods: To produce TP-specific scFv (single-chain Fv) by phage display, mice were immunized using synthetic TP-peptide contains $57{\sim}80^{th}$ amino acid residues of TP domain. After isolation of mRNA of heavy-variable region ($V_H$) and light-chain variable region ($V_L$) from the spleen of the immunized mouse, DNA of $V_H$ and $V_L$ were obtained by RT-PCR and joined by a DNA linker encoding peptide (Gly4Ser)3 as a scFv DNA fragments. ScFv DNA fragments were cloned into a phagemid vector. scFv was expressed in E.coli TG1 as a fusion protein with E tag and phage gIII. To select the scFv that has specific affinity to TP-peptide from the phage-antibody library, we used two cycles of panning and colony lift assay. Results: The TP-peptide-specific scFv was isolated by selection process using TP-peptide as an antigen. Selected scFv had 30 kDa of protein size and its nucleotide sequences were analyzed. Indirect- and competitive-ELISA revealed that the selected scFv specifically recognized both TP-peptide and the HBV DNA polymerase. Conclusion: The scFv that recognizes the TP domain of the HBV DNA polymerase was isolated by phage display.

• The Korean Journal of Blood Transfusion
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• v.23 no.2
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• pp.173-179
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• 2012

We report on two cases of anti-$Jk^a$, whose reactivity disappeared on an antibody identification test using enzyme-treated red cells. One of two patients was a 72-year-old female with cirrhosis of the liver and colon cancer, and the other was a 55-year-old female with known MDS and incomplete Behcet's disease. Results of an antibody identification test using a LISS/Coombs gel card (DiaMed AG) showed negative to one positive with red cells having the $Jk^a$ antigen; however, all reactions using the enzyme-treated cells showed negative results, which was unexpected. The patients' RBC phenotype was Jk(a-b+). We obtained positive results in reactions of enzyme-treated $Jk^a+$ cells and EDTA using a patient's serum and proved that the cause of the negative reaction might be complement-related.

• Korean Journal of Veterinary Research
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• v.15 no.1
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• pp.57-67
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• 1975

The poxvirus group is considered to be a typical cytoplasmic inclusion forming virus. Every poxvirus has been reported to produce only one kind of inclusion in the infected tissues. A vague concept that inclusions of poxviruses are eosinophilic or acidophilic has prevailed. Although many papers and theories about the nature of the inclusion have been presented, most of them are not quite convincing on the point of the relations with virus multiplication, and an analysis of papers published showed that there seem to be many discrepancies in the descriptions of the nature of the poxvirus inclusions. Comparative studies on host-virus interaction with cowpox, orf, swinepox and fowlpox viruses which selected from each Group (I-IV) of poxviruses were performed from the morphological and virological standpoints. At first, in cowpox virus-FL cell system, as a comparative model, cytoplasmic inclusion, nucleic acid metabolism by autoradiography and detection of viral antigen by immunofluorescence were studied and obtained the results as follows: 1. The focus-like cytopathic effect (CPE) at early stage developed to entire culture at terminal stage of infection, and also the developing status of CPE was correlated to viral doses for inoculation. Two kinds of cytoplasmic inclusions which named A and B type were easily observed by Giemsa, hematoxylin-eosin (H & E) and May-Greenwald Giemsa (MGG) stainings in the infected cells. The B type inclusions were formed at early stage of infection and the A type inclusions were produced subsequently the B type formation. The B type which common type inclusion in poxviruses was a small compact or aggregate at early stage and developed to a large diffuse body at terminal stage of infection. On the other hand, the A type inclusion which depend upon the kind of virus was appeared as round and discrete shape, and its size and number was increased gradually during the culture period. It was characteristic to form distinct halos around the both types of inclusions in acid fixed, H & E stained preparations of infected cultures. The B type inclusion was always positive in Feulgen reaction and showed as DNA containing body but the A type inclusion was not. 2. In the relationship between inclusion and DNA metabolism of infected cells by the qualitative autoradiography using 3H-thymidine, the appearance of silver grains was coincided with B type inclusion but not with A type inclusion. This showed that the DNA synthesis was proceeded in all B type inclusions except those in the terminal stage with a diffuse form. This suggested that the B type inclusions are only sites of DNA synthesis and this was proceeded after the cell infection independently. The activity of DNA synthesis of the inclusions was nearly the same as that of the nucleic of normal cells and non-inclusion bearing cells. and non-inclusion bearing cells. Regardless of the size of the degree of DNA synthesis of the B type inclusion, inclusion bearing cells all showed remarkable suppression of nuclear DNA synthesis. 3. By the direct fluorescent antibody technique viral antigen in infected cells was detected. The B type inclusions have been proved to contain a great deal of viral antigen, whereas the basic substance of A type inclusion did not show antigenicity except the round edge. It was suggested that the round edge fluorescence might be caused by the glare of cytoplasmic viral antigen which pushed out and concentrated by the A type inclusion development. 4. Hemorrhagic red pock formations on chorioallantoic membrane of embryonated chicken egg had proved the characteristic of used viral strain. 5. By the above studies on the nature of two types of inclusions and the role they play in virus multiplication, it was concluded that the B type inclusion must be the site of the synthesis of viral DNA and protein as well as the site of the virus.

• Journal of Veterinary Clinics
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• v.30 no.5
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• pp.333-338
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• 2013

We investigated the prevalence of feline panleukopenia virus (FPV) in stray and household cats in different regions of Seoul, Republic of Korea. Blood samples were collected from a total of 200 cats (100 stray cats and 100 household cats) and examined by polymerase chain reaction (PCR). The overall prevalence of FPV was 2%. Among test-positive cats, 3% (3/100) were stray cats and 1% (1/100) was a household cat. The incidence of FPV was higher in juvenile cats (< 1 year, 1.5%) than in adult cats (> 1-year-old, 0.5%). The FPV-positive rates of healthy infected cats and sick cats were 1.9% (3/156) and 2.2% (1/44), respectively. We found the positive rate of vaccinated and unvaccinated cats to be 1.3% (1/77) and 2.4% (3/123), respectively. Unlike antibody tests, FPV antigen tests detected current infections in stray and household cats. Therefore, these tests can help in disease diagnosis and treatment. To our knowledge, our study is the first to survey the prevalence of FPV in different cat populations across Seoul. We found a high prevalence of FPV infection in stray and juvenile cats. Therefore, proper vaccination and surveillance are important to prevent FPV outbreaks.