• Journal of Welding and Joining
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• v.26 no.1
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• pp.56-62
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• 2008

In this paper, elastic and plastic adhesion index was very important in deciding adhesive characteristics and varying elastic and plastic index, dimensionless load and pull-off force were analyzed and simulated. Finally, using AFM, experimental surface roughness parameters of substrates and pull-off force between tip and substrates were produced. Using these values, pull-off forces were calculated and were compared with experimental pull-off forces. Through simulation and experiment, it was found that interaction of asperity also had very important influence on adhesive contact.

• International Journal of Oral Biology
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• v.33 no.2
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• pp.45-50
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• 2008

Treponema denticola is the best studied oral spirochete and numerous studies have shown that it is strongly associated with periodontitis and expresses several putative virulence factors. In this study, we report on a surface protein of T. denticola, Td92, which is homologous to Tp92 of Treponema pallidum, an agent of syphilis. Immunofluorescence assay and immunogold labeling with anti-Td92 Ab revealed that Td92 had surface-exposed epitopes. And Td92 was capable of binding to fibronectin and KB cells, an oral epithelial cell line. In addition, Td92 could enter the KB cells. These results indicate that Td92 is a fibronectin-binding protein which can bind to and internalize into the host cells, facilitating the virulence of T. denticola.

• Proceedings of the Korean Society of Veterinary Pathology Conference
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• 2003.10a
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• pp.30-30
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• 2003

Escherichia coli strains associated with diarrhea have been divided into the following six major categories on the basis of pathogenic mechanisms: enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC), enteropathogenic E. coli (EPEC), enterohemorrhagic E. coli (EHEC), enteroaggregative E. coli (EAggEC) and diffusely adherent E. coli (DAEC).$\^$15,18/ EPEC, EAggEC, and DAEC strains were classified by their ability to produce distinct patterns of adherence to cultured epithelial cells in virto: localized (LA), aggregative (AA), and diffuse (DA) adherence. The objective of this study was to investigate the relationship of the adherence patterns with AIDA-positive E. coli isolated from diarrheic pigs. (omitted)

• Journal of Microbiology
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• v.44 no.5
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• pp.548-555
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• 2006

FaeG is the key factor in the infection process of K88ad enterotoxigenic Escherichia coli (ETEC) fimbrial adhesin. In an attempt to determine the possibility of expressing recombinant FaeG with immunogenicity for a new safe and high-production vaccine in E. coli, we constructed the recombinant strain, BL21 (DE3+K88), which harbors an expression vector with a DNA fragment of faeG, without a signal peptide. Results of 15% SDS-polyacrylamide slab gel analysis showed that FaeG can be stably over-expressed in BL21 (DE3+K88) as inclusion bodies without FaeE. Immunoglobulin G (IgG) and M (IgM) responses in pregnant pigs, with boost injections of the purified recombinant FaeG, were detected 4 weeks later in the sera and colostrum. An in vitro villius-adhesion assay verified that the elicited antibodies in the sera of vaccinated pigs were capable of preventing the adhesion of K88ad ETEC to porcine intestinal receptors. The protective effect on the mortality rates of suckling piglets born to vaccinated mothers was also observed one week after oral challenge with the virulent ETEC strain, $C_{83907}$ (K88ad, $CT^+,\;ST^+$). The results of this study proved that the adhesin of proteinaceous bacterial fimbriae or pili could be overexpressed in engineered E. coli strains, with protective immune responses to the pathogen.

• Korean Journal of Veterinary Research
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• v.38 no.4
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• pp.837-842
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• 1998

The present study describes the effectiveness of egg yolk antibodies (IgY) against enteric colibacillosis and edema disease in piglets. The antibodies were gained from the egg yolk of hens immunized with k88, k99, 987p fimbrial adhesin and heat-labile toxin antigens of enterotoxigenic Escherichia coli (ETEC). Orally-administered egg yolk antibodies solution protected against experimental challenge with ETEC $K88^+$ and $k99^+$ strains in neonatal piglets and mice. In field trial, a total of 598 diarrheal piglets were orally treated with 3ml of antibody once a day to determine for the therapeutic effect. Of them, 582 (97.3%) piglets were recovered from diarrhea in 3 days. We also experimentally treated with the egg yolk antibodies twice a day for 5 consecutive days for 94 weaning piglets with edema disease for the determination of therapeutic effects. Seventy four piglets (78.7%) were recovered from clinical edema signs. Theses findings indicate that egg yolk antibodies against k88, k99, 987p and LT of ETEC are useful source of passive immunity for enteric colibacillosis and edema disease of piglets.

• Journal of Microbiology and Biotechnology
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• v.27 no.10
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• pp.1844-1854
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• 2017

Trichomonas vaginalis is a pathogen that triggers severe immune responses in hosts. T. vaginalis ${\alpha}$-actinin 2 ($Tv{\alpha}$-actinin 2) has been used to diagnose trichomoniasis. $Tv{\alpha}$-actinin 2 was dissected into three parts; the N-terminal, central, and C-terminal portions of the protein (#1, #2, and #3, respectively). Western blot of these $Tv{\alpha}$-actinin 2 proteins with pooled patients' sera indicated that #2 and #3, but not #1, reacted with those sera. Immunofluorescence assays of two different forms of T. vaginalis (trophozoites and amoeboid forms), using anti-$Tv{\alpha}$- actinin 2 antibodies, showed localization of $Tv{\alpha}$-actinin 2 close to the plasma membranes of the amoeboid form. Fractionation experiments indicated the presence of $Tv{\alpha}$-actinin 2 in cytoplasmic, membrane, and secreted proteins of T. vaginalis. Binding of fluorescence-labeled Trichomonas to vaginal epithelial cells and prostate cells was decreased in the antibody blocking experiment using anti-$Tv{\alpha}$-actinin 2 antibodies. Pretreatment of T. vaginalis with anti-$rTv{\alpha}$-actinin 2 antibodies also resulted in reduction in its cytotoxicity. Flow cytometry, ligand-binding immunoblotting assay, and observation by fluorescence microscopy were used to detect the binding of recombinant $Tv{\alpha}$-actinin 2 to human epithelial cell lines. Specifically, the truncated N-terminal portion of $Tv{\alpha}$-actinin 2, $Tv{\alpha}$-actinin 2 #1, was shown to bind directly to vaginal epithelial cells. These data suggest that ${\alpha}$-actinin 2 is one of the virulence factors responsible for the pathogenesis of T. vaginalis by serving as an adhesin to the host cells.

• Journal of Microbiology and Biotechnology
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• v.20 no.4
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• pp.789-797
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• 2010

The limited information on differential gene expression in the different serotypes of Actinobacillus pleuropneumoniae has significantly hampered the research on the pathogenic mechanisms of this organism and the development of multivalent vaccines against A. pleuropneumoniae infection. To compare the gene expressions in the A. pleuropneumoniae strains CVCC259 (serotype 1) and CVCC261 (serotype 3), we screened the differentially expressed genes in the two strains by performing representational difference analysis (RDA). Northern blot analyses were used to confirm the results of RDA. We identified 22 differentially expressed genes in the CVCC259 strain and 20 differentially expressed genes in the CVCC261 strain, and these genes were classified into 11 groups: (1) genes encoding APX toxins; (2) genes encoding transferrin-binding protein; (3) genes involved in lipopolysaccharide (LPS) biosynthesis; (4) genes encoding autotransporter adhesin; (5) genes involved in metabolism; (6) genes involved in the ATP-binding cassette (ABC) transporter system; (7) genes encoding molecular chaperones; (8) genes involved in bacterial transcription and nucleic acid metabolism; (9) a gene encoding protease; (10) genes encoding lipoprotein/membrane protein; and (11) genes encoding various hypothetical proteins. This is the first report on the systematic application of RDA for the analysis of differential gene expression in A. pleuropneumoniae serotypes 1 and 3. The determination of these differentially expressed genes will serve as an indicator for future research on the pathogenic mechanisms of A. pleuropneumoniae and the development of a multivalent vaccine against A. pleuropneumoniae infection.

• Journal of Microbiology and Biotechnology
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• v.28 no.10
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• pp.1723-1729
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• 2018

The aim of this work is to investigate the protective efficacy of emodin, an active, naturally-occurring anthraquinone derivative of several traditional Chinese herbs, against Brucella abortus infection in macrophages. Brucella were incubated with different concentrations of emodin and showed that bacterial survival rates were markedly reduced in a dose-dependent manner at increasing incubation time points. Through bacterial infection assay, the highest non-cytotoxic concentration of emodin demonstrated attenuated invasion of Brucella into macrophages, however it did not inhibit the growth of these pathogens within the host cells. On the other hand, emodin effectively decreased the number of bacteria that adhered to host cells, which indicated its potential as an anti-adhesin agent. Furthermore, using immunoblotting and FACS assay for detecting MAPK signaling proteins and F-actin polymerization, respectively, the results showed that the emodin-incubated cells displayed modest reduction in the phosphorylation levels of ERK1/2 and inhibition of F-actin polymerization as compared to control cells. These findings indicate the potential use of emodin as a naturally-occurring alternative method for the prevention of animal brucellosis although this requires confirmation of safe clinical doses.

• Journal of Microbiology and Biotechnology
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• v.27 no.2
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• pp.262-270
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• 2017

Yersinia enterocolitica is a well-known foodborne pathogen causing gastrointestinal infections worldwide. The strain Y. enterocolitica FORC_002 was isolated from the gill of flatfish (plaice) and its genome was sequenced. The genomic DNA consists of 4,837,317 bp with a GC content of 47.1%, and is predicted to contain 4,221 open reading frames, 81 tRNA genes, and 26 rRNA genes. Interestingly, genomic analysis revealed pathogenesis and host immune evasion-associated genes encoding guanylate cyclase (Yst), invasin (Ail and Inv), outer membrane protein (Yops), autotransporter adhesin A (YadA), RTX-like toxins, and a type III secretion system. In particular, guanylate cyclase is a heat-stable enterotoxin causing Yersinia-associated diarrhea, and RTX-like toxins are responsible for attachment to integrin on the target cell for cytotoxic action. This genome can be used to identify virulence factors that can be applied for the development of novel biomarkers for the rapid detection of this pathogen in foods.

• Journal of Microbiology and Biotechnology
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• v.31 no.8
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• pp.1088-1097
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• 2021

Grouper nervous necrosis virus (GNNV) infection causes mass grouper mortality, leading to substantial economic loss in Taiwan. Traditional methods of controlling GNNV infections involve the challenge of controlling disinfectant doses; low doses are ineffective, whereas high doses may cause environmental damage. Identifying potential methods to safely control GNNV infection to prevent viral outbreaks is essential. We engineered a virus-binding bacterium expressing a myxovirus resistance (Mx) protein on its surface for GNNV removal from phosphate-buffered saline (PBS), thus increasing the survival of grouper fin (GF-1) cells. We fused the grouper Mx protein (which recognizes and binds to the coat protein of GNNV) to the C-terminus of outer membrane lipoprotein A (lpp-Mx) and to the N-terminus of a bacterial autotransporter adhesin (Mx-AIDA); these constructs were expressed on the surfaces of Escherichia coli BL21 (BL21/lpp-Mx and BL21/Mx-AIDA). We examined bacterial surface expression capacity and GNNV binding activity through enzyme-linked immunosorbent assay; we also evaluated the GNNV removal efficacy of the bacteria and viral cytotoxicity after bacterial adsorption treatment. Although both constructs were successfully expressed, only BL21/lpp-Mx exhibited GNNV binding activity; BL21/lpp-Mx cells removed GNNV and protected GF-1 cells from GNNV infection more efficiently. Moreover, salinity affected the GNNV removal efficacy of BL21/lpp-Mx. Thus, our GNNV-binding bacterium is an efficient microparticle for removing GNNV from 10‰ brackish water and for preventing GNNV infection in groupers.