• Korean Journal of Microbiology
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• v.41 no.4
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• pp.275-280
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• 2005

Anthrax is an infectious disease caused by the gram-positive bacterium, Bacillus anthracis. Anthrax toxin is a tripartite toxin comprising of protective antigen (PA), lethal factor (LF) and edema factor (EF). PA is the receptor-binding component, which facilitates the entry of LF or EF onto the cytosol. LF is a zinc-dependent metalloprotease, which is a critical virulence factor in cytotoxicity of infected animals. Therefore, it is of interest to develop its potent inhibitors for the neutralization of anthrax toxin. The first step to identify the inhibitors is the development of a rapid, sensitive, and simple assay method with a high-throughput ability. Much efforts have been concentrated on the preparation of powerful assays and on the screening of inhibitors using these system. In the present study, we have tried to construct anthrax lethal factor in yeast expression system to prepare cell-based high-throughput assay system. Here, we have shown the results covering the construction of a new vector system, subcloning of LF gene, and the expression of target gene. Our results are first trial to express LF gene in eukaryote and provide the basic steps in design of cell-based assay system.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.8
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• pp.1065-1071
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• 2013

The swine leukocyte antigen class II molecules are possibly associated with the induction of protective immunity. The study described here was to investigate the relationship between polymorphisms in exon 2 of the swine DQA gene and piglet diarrhea. This study was carried out on 425 suckling piglets from three purebred pig strains (Large White, Landrace and Duroc). The genetic diversity of exon 2 in swine DQA was detected by PCR-SSCP and sequencing analysis, eight unique SSCP patterns (AB, BB, BC, CC, CD, BD, BE and DD) representing five specific allele (A to E) sequences were detected. Sequence analysis revealed 21 nucleotide variable sites and resulting in 12 amino acid substitutions in the populations. A moderate level polymorphism and significant deviations from Hardy-Weinberg equilibrium of the genotypes distribution were observed in the populations (p<0.01). The association analysis indicated that there was a statistically significant difference in the score of piglet diarrhea between different genotypes, individuals with genotype CC showed a lower diarrhea score than genotypes AB ($0.98{\pm}0.09$), BB ($0.85{\pm}0.77$) and BC ($1.25{\pm}0.23$) (p<0.05), and significantly low than genotype BE ($1.19{\pm}0.19$) (p<0.01), CC genotype may be a most resistance genotype for piglet diarrhea.

• Korean Journal of Veterinary Pathology
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• v.6 no.1
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• pp.19-26
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• 2002

The purpose of this study was to investigate to potective effects against porcine epidemic diarrhea virus (PEDV) infection in piglets by administration of the PEDV antiserum orally at 2 hrs, 24hrs and 36hrs after birth. six piglets administered the antiserum were experimentally infected with PEDV at five-day-old. Control group were four piglets infected with PEDV only. Serum antibody titers against PEDV were examined by serum neutralization (SN) test, dectection for PEDV or PEDV antigen from feces and small intestines was tested by reverse transcription-polymerase chain reaction (RT-PCR) and indirect immunoflurescence (IFA). The results obtained were as follows; 1. The piglets administered the PEDV antiserum showed higher antibody titers than those of control group and sustained during the experimental period. 2. The detection rate of PEDV in feces and small intestines by RT-PCR were 26.2% and 16.7% in PEDV antiserum treated group and 48.1 % and 75.0% in control group, respectively. 3. The detection rate of PEDV antigen in the small intestine by IFA were 0% in PEDV antiserum treated group and 50.0% in control group, respectively. It was concluded that oral administration of antiserum against PEDV to piglets was effective in preventing PEDV infection.

• Korean Journal of Veterinary Pathology
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• v.6 no.1
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• pp.11-18
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• 2002

The purpose of this study was to investigate to potective effects against transmissible gastyoenteritis virus (TGEV) infection in piglets by administration of the TGEV antiserum orally at 5 hrs, 24hrs and 36hrs after birth. five piglets administered the antiserum were experimentally infected with TGEV at four-day-old. Control group were four piglets infected with TGEV only. Serum antibody titers against TGEV were examined by serum neutralization(SN) test, dectection for TGEV or TGEV antigen from feces and small intestines was tested by reverse transcrption-polymerase chain reaction (RT-PCR) and indirect immunoflurescence (IFA). The results obtained were as follows; 1. The piglets administered the TGEV antiserum showed higher antibody titers than those of control group and sustained during the experimental period. 2. The detection rate of TGEV in feces and small intestines by RT- PCR were 24.5% and 20.0% in TGEV antiserum treated group and 44.0% and 75.0% in control group, respectively. 3 The detection rate of TGEV antigen in the small intestine by IFA were 26.7% in TGEV antiserum treated group and 75.0% in control group, respectively. It was concluded that oral administration of antiserum against TGEV to piglets was effective in preventing TGEV infection.

• Journal of Acupuncture Research
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• v.23 no.2
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• pp.91-102
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• 2006

Objectives : Ulmus davidiana Planch (UD) has long been known to have anti-inflammatory and protective effects on damaged tissue, inflammation and bone among other functions. Methods : This study was undertaken to address whether the water extract of the bark of UD could modulate proliferation of mouse osteoclasts in vitro and to investigate its effect on cyclooxygenase-2 (COX-2), which converts arachidonic acid to prostaglandin E2 (PGE2) and is highly expressed in osteoclasts. Mouse osteoclasts were tested in vitro for growth inhibition, proliferation cell nuclear antigen expression, and COX-2 activity and expression after treatment with UD extract. Results : Its effects were compared with those of indomethacin (a nonselective COX inhibitor) and celecoxib (a selective COX-2 inhibitor) by Cell viability assay, Cell cycle analysis, Immunohistochemical analysis of PCNA expression, Western blot analysis and PGE2 Enzyme immunoassay (EIA). UD demonstrated a strong growth inhibitory action in both tested osteoclasts cells. The IC50s were $10\;{\mu}g/ml$ for UD, $6\;{\mu}M$ for celecoxib and $42\;{\mu}M$ for indomethacin. UD, as well as celecoxib and indomethacin, suppressed proliferation cell nuclear antigen expression and PGE2 synthesis in osteoclasts. UD inhibited COX-2 expression, whereas celecoxib inhibited COX-2 activity directly. Conclusion : UD selectively and effectively inhibits osteoclasts cell growth in vitro. Inhibitory action of PGE2 synthesis via suppression of COX-2 expression may be responsible for its anti-inflammatory activity.

• Parasites, Hosts and Diseases
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• v.50 no.1
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• pp.45-51
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• 2012

Fascioliasis is one of the public health problems in the world. Cysteine proteinases (CP) released by $Fasciola$ $gigantica$ play a key role in parasite feeding, migration through host tissues, and in immune evasion. There has been some evidence from several parasite systems that proteinases might have potential as protective antigens against parasitic infections. Cysteine proteinases were purified and tested in vaccine trials of sheep infected with the liver fluke. Multiple doses (2 mg of CP in Freund's adjuvant followed by 3 booster doses 1 mg each at 4 week intervals) were injected intramuscularly into sheep 1 week prior to infect orally with 300 $F.$ $gigantica$ metacercariae. All the sheep were humanely slaughtered 12 weeks after the first immunization. Changes in the worm burden, ova count, and humoral and cellular responses were evaluated. Significant reduction was observed in the worm burden (56.9%), bile egg count (70.7%), and fecel egg count (75.2%). Immunization with CP was also found to be associated with increases of total IgG, $IgG_1$, and $IgG_2$ ($P$<0.05). Data showed that the serum cytokine levels of pro-inflammatory cytokines, IL-12, IFN-${\gamma}$, and TNF-${\alpha}$, revealed significant decreases ($P$<0.05). However, the anti-inflammatory cytokine levels, IL-10, TGF-${\beta}$, and IL-6, showed significant increases ($P$<0.05). In conclusion, it has been found that CP released by $F.$ $gigantica$ are highly important candidates for a vaccine antigen because of their role in the fluke biology and host-parasite relationships.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.4
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• pp.362-366
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• 2005

Actinobacillus pleuropneumoniae is an important pig pathogen, which is responsible for swine pleuropneumonia, a highly contagious respiratory infection. To develop subunit vaccines for A. pleuropneumoniae infection, the Apx toxin genes, apxI and apxII, which are thought to be important for protective immunity, were expressed in Saccharomyces cerevisiae, and the induction of immune responses in mice was examined. The apxI and apxII genes were placed under the control of a yeast hybrid ADH2-GPD promoter (AG), consisting of alcohol dehydrogenase II (ADH2) and the GPD promoter. Western blot analysis confirmed that both toxins were successfully expressed in the yeast. The ApxIA and ApxIIA-specific IgG antibody response assays showed dose dependent increases in the antigen-specific IgG antibody titers. The challenge test revealed that ninety percent of the mice immunized with ApxIIA or a mixture of ApxIA and ApxIIA, and sixty percent of mice immunized with ApxIA survived, while none of those in the control groups survived longer than 36 h. These results suggest that vaccination of the yeast ex­pressing the ApxI and ApxII antigens is effective for the induction of protective immune responses against A. pleuropneumoniae infections in mice.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.3
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• pp.1813-1817
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• 2013

Background: Multiple etiologic factors are suspected to cause gastric cancer, the most important of which is infection with virulent types of Helicobacter pylori. Materials and Methods: We have compared 102 gastric cancer patients with 122 non-ulcer, non-cancer dyspeptic patients. Gastric specimens were evaluated for H. pylori infection by tissue-based detection methods. Patient sera underwent antigen-specific ELISA and western blotting using a Helicoblot 2.1 kit and antibody responses to various H. pylori antigens were assessed. Results: The absolute majority (97-100%) of both groups were H. pylori seropositive. Multivariate regression analysis demonstrated serum antibodies to the low molecular weight 35kDa protein to be protective and reduce the risk of gastric cancer by 60% (OR:0.4; 95%CI:0.1-0.9). Conversely, seroreactivity to the 89kDa (VacA) protein was significantly higher in gastric cancer patients (OR:2.7; 95%CI:1.0-7.1). There was a highly significant association (p<0.001) between seroreactivity to the 116kDa (CagA) and 89kDa (VacA) proteins, and double positive subjects were found at nearly five fold (OR:4.9; 95%CI:1.0-24.4) enhanced risk of gastric cancer as compared to double negative subjects. Conclusions: Seroreactivity to H. pylori low (35kDa) and high (116kDa/89kDa) molecular weight antigens were respectively revealed as protective and risk indicators for gastric cancer.

• The Journal of Korean Medicine
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• v.22 no.2
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• pp.84-93
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• 2001

This research was to find out the protective and healing effects of both the Jiguyangwi-tang and the Gamijiguyangwi-tang on the gastric mucosa injuries by cyclophosphamide. At first, Jiguyangwi-tang and Gamijiguyangwi-tang extract were administered to the mice before one week, and then integral administration of those two drugs(each herbal extract and cyclophosphamide) were followed for another one week, respectively. After finishing those treatments, I have examined the state of the both ulcer and inflammation on the damaged gastric mucosa cell and watched the level of proliferating cell nuclear antigen(PCNA), Bcl-2, and apoptosis. These results were as follows, 1. Gastric mucosa inflammation have more significantly reduced in groups of integral administration of Jiguyangwi-tang plus cyclophosphamide, Gamijiguyangwi-tang plus cyclophosphamide and cyclophosphamide then in group of single cyclophosphamide. 2. Gastric ulcer have been reduced in groups of integral administration of Jiguyangwi-tang plus cyclophosphamide, Gamijiguyangwi-tang plus cyclophosphamide and cyclophosphamide then in group of single cyclophosphamide. But the significance have not shown. 3. PCNA level have more significantly elevated in integral administration of Jiguyangwi-tang plus cyclophosphamide and Gamijiguyangwi-tang plus cyclophosphamide and cyclophosphamide then in group of single cyclophosphamide. 4. The significance of both apoptosis induction and bcl-2level have not noticed among all groups. 5. Between Jiguyangwi-tang and Gamijiguyangwi-tang, the differance of effect was not admitted in statistically From these results, it is suggested that Jiguyangwi-tang and Gamijiguyangwi-tang are useful medicines in protecting gastric inflammation and ulcer, that is gastrointestinal side-effect of cyclophosphamide. The preventing effect of Jiguyangwi-tang and Gamijiguyangwi-tang may be through the 'affecting the period of cell division', but not the inhibition of apoptosis.

• Applied Biological Chemistry
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• v.13 no.1
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• pp.81-85
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• 1970

In this studies, we isolated a kind of protein from Alisma Canaliculatum by the saline extraction. This protein was found to have a strong protective effects on the denaturation of ${\alpha}-chymotrypsin$ in the solution state. The obtained important results during the studies were as follows, 1. This protein was never hydrolyzed by the ${\alpha}-chymotrypsin$. 2. The denaturation of ${\alpha}-chymotrypsin$ was strongly protected by this sample protein. 3. Isoelectric point of this sample was about 4.7. 4. This sample protein was determined as an antigen but very weak antigenicity was indicated on rabbit.