• Microbiology and Biotechnology Letters
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• v.46 no.3
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• pp.261-268
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• 2018

Differences in host ethnicities and geographical distributions may influence the genetic variation and pathogenesis of Helicobacter pylori strains, particularly with respect to those with a high risk of gastric cancer and in Asian Enigma regions. We simultaneously identified H. pylori virulence-associated genes involved in inflammation and cell damage in Thai and Korean dyspeptic patients. The virulence-associated gene cagA, cagA genotypes (East Asian and Western type cagA), vacA genotypes (s- and m-), oipA, and sabA were detected in Thai and Korean dyspeptic patients by polymerase chain reaction (PCR), real-time PCR, and DNA sequence analysis. Comparisons between the two regions showed that cagA, East Asian type cagA, and vacA s1/m1 in Korean dyspeptic patients occurred at rates of 100%, 86.67%, and 88.89%, respectively (p < 0.05). The oipA- and sabA-positive samples were significantly more predominant in the Korean population (95.56%, 91.11%) than in the Thai population (32%, 34%). DNA sequence analysis revealed differences in the patterns of cytosine-thymine dinucleotide repeats of oipA and sabA among the two populations of dyspeptic patients. Our results indicate that the H. pylori strains detected in the two regions were divergent, and strains colonizing the Korean dyspeptic patients may be more virulent than those in the Thai population. Our data may help explain H. pylori pathogenesis in Asian Enigma areas with a low gastric cancer incidence. However, other factors involving H. pylori infection in these two regions should be further analyzed.

• IMMUNE NETWORK
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• v.3 no.4
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• pp.302-309
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• 2003

Background: CTLA4 (CD152), which is expressed on the surface of T cells following activation, has a much higher affinity for B7 molecules comparing to CD28, and is a negative regulator of T cell activation. In contrast to stimulating and agonistic capabilities of monoclonal antibodies specific to CTLA-4, CTLA4Ig fusion protein appears to act as CD28 antagonist and inhibits in vitro and in vivo T cell priming in variety of immunological conditions. We've set out to confirm whether inhibition of the CD28-B7 costimulatory response using a soluble form of human CTLA4Ig fusion protein would lead to persistent inhibition of alloreactive T cell activation. Methods: We have used CHO-$dhfr^-$ cell-line to produce CTLA4Ig fusion protein. After serum free culture of transfected cell line we purified this recombinant molecule by using protein A column. To confirm characterization of fusion protein, we carried out a series of Western blot, SDS-PAGE and silver staining analyses. We have also investigated the efficacy of CTLA4Ig in vitro such as mixed lymphocyte reaction (MLR) & cytotoxic T lymphocyte (CTL) response and in vivo such as experimental autoimmune encephalomyelitis (EAE), graft versus host disease (GVHD) and skin-graft whether this fusion protein could inhibit alloreactive T cell activation and lead to immunosuppression of activated T cell. Results: In vitro assay, CTLA4Ig fusion protein inhibited immune response in T cell-specific manner: 1) Human CTLA4Ig inhibited allogeneic stimulation in murine MLR; 2) CTLA4Ig prevented the specific killing activity of CTL. In vivo assay, human CTLA4Ig revealed the capacities to induce alloantigen-specific hyporesponsiveness in mouse model: 1) GVHD was efficiently blocked by dose-dependent manner; 2) Clinical score of EAE was significantly decreased compared to nomal control; 3) The time of skin-graft rejection was not different between CTLA4Ig treated and control group. Conclusion: Human CTLA4Ig suppress the T cell-mediated immune response and efficiently inhibit the EAE, GVHD in mouse model. The mechanism of T cell suppression by human CTLA4Ig fusion protein may be originated from the suppression of activity of cytotoxic T cell. Human CTLA4Ig could not suppress the rejection in mouse skin-graft, this finding suggests that other mechanism except the suppression of cytotoxic T cell may exist on the suppression of graft rejection.

• Journal of Periodontal and Implant Science
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• v.33 no.2
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• pp.289-299
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• 2003

The autogenous free gingival graft is the most predictable procedure currently used to increase the width of　the attached gingiva in periodontics. But the major disadvantage of the procedure is to create the multiple surgical wounds at both a donor site and a recipient site. The other problem is the limited amount of available　graft material in oral cavity. Therefore, recent researches have been focused to develop the biomaterial to substitute the autogenous gingival tissue. The purpose of this study was to evaluate the histologic healing after　grafting of bilayer artificial dermis, compared to the free gingival graft. Four non-smoking subjects (mean age, 32.5 years) in systemically healthy state and good oral hygiene were selected according to their particular needs for correction of mucogingival problems as suggested by Nabers(1966). The recipient sites were prepared through the procedure for the free gingival graft and were grouped according to the graft materials: Experimental group(n=5) - bilayer artifcia1 dermis ($Terudermis^{(R)}$; Terumo Co. Japan) and Control group(n=6) - free gingival graft with autogenous palatal mucosa. Biopsies were harvested at 1,2,3 and 6 weeks postsurgery to evaluate histologically. At the third week in the experimental group and at the second week of in the control group, the grafts has been clinically stabilized on the recipient bed and the graft border has been blended into the surrounding tissue. In the experimental group after 1 week of grafting, the epithelial migration from the adjacent tissue to graft material was seen and after 3 weeks of grafting, the : nflmmation decreased, collagen layer of the artificial dermis was lost and the basement membrane of epithelium was formed. After 6 weeks of grafting, both groups demonstrated orthokeratinized epithelium and increased thickness of epithelial tissue and the rete peg formation, similar to the adjacent tissue, Histologic evaluation revealed a biologic acceptance and incorporation of the collagen layers of the graft tissue to the host tissue, without foreign body reaction. In conclusion, a bilayer artificial dermis is essentially similar to autogeneous free gingival graft in the correction of mucogingival problems, and has the advantages of decreased patient morbidity (no donor site) and availability of abundant amounts of graft material when needed.

• Korean Journal Plant Pathology
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• v.3 no.3
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• pp.223-235
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• 1987

Samples showing yellow mosaic symptom of Lilium spp. with necrotic fleck, stunting, malformation, and colour breaking were collected from lily-growing areas in the southern part of Korea. Two viruses were distinguished under a electron microscope and their host range, serological reaction, stability in sap, type of aphid transmission, and relations with cells and tissues were examined. Broad bean wilt virus (BBWV) was transmitted by sap-inoculation to 23 plant species in 8 families and by the aphid, Myzus persicae. This virus was inactivated after 10 min at 70C, at dilution of $10^{-3}$, and after 6 days at about 20C. Electron microscopic examination of purified preparation showed that the virus is spherical particle of 28nm in diameter. The virus reacted positively with BBWV-antiserum in agar gel diffusion test. In ultrathin sections of BBWV infected tissues, large aggregates or crystalline array of virus particles and vesicular body were found in the cytoplasm, vacuole, and nucleus of mesophyll cells. Cucumber mosaic virus (CMV) was transmitted by sap-inoculation. Electron microscopic examination of its purified preparation showed spherical particles of 30nm in diameter. The virus reacted positively with CMV-Y strain-antiserum in agar gel diffusion test. In ultrathin sections of CMV infected tissues, crystalline array of virus particles were found in the vacuole and a large number 0f virus particles were found in the cytoplasm and the plasmodesmata of mesophyll cells. When each of these viruses was retransmitted to Lilium tigrinum. L. concolor, and L. auratum, BBWV induced slight symtoms and colour breaking, but CMV induced yellowing mosaic or necrotic fleck.

• Parasites, Hosts and Diseases
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• v.22 no.2
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• pp.229-237
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• 1984

The migration and distribution pattern of spargana in mouse body was observed after experimental infection through mouth. The spargana were obtained from the snake, Natris tigrina lateralis, caught in Hoengseong-gun, Kangwon-do. A total of 28 male mice (ICR strain), 21∼259 in body weight, were fed each with 5 scolices (and necks) of spargana and killed after 10 minutes to 14 days. Systemic autopsy was performed on each mouse to recover the spargana. The results are as follows: 1. The spargana were found to penetrate into the stomach or duodenal wall of mice as early as 10 minutes after infection. They completed the penetration within 30 minutes and appeared in abdominal cavity. It was observed that spargana did not migrate tangentially along the gut wall but directly perforated the wall. 2. After 1 hour to 1 day the majority of spargana distributed in abdominal cavity of mice except a few which migrated to muscles or subcutaneous tissues. 3. It was within 7 days that nearly all of the spargana migrated to subcutaneous tissues. Out of total 28 in number found from subcutaneous tissues, 13 distributed around neck region, 12 around trunk and other 3 on head of mice and the most common sites were submandibular and subscapular areas. There was nearly no host tissue reaction to migrating spargana. 4. The initial length of spargana given was 4 mm in average but it increased to 12 mm after 7 days and to 35 mm after 14 days. The results suggest that spargana orally given to mice penetrate the gut wall within 30 minutes followed by escaping into abdominal cavity, and after passing through thoracic cavity or abdominal wall they anally Localize in subcutaneous tissues chieay around neck region within 7 days.

• Biomedical Science Letters
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• v.11 no.3
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• pp.259-266
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• 2005

Mast cells and goblet cells have been known to protect the host against parasites. In this study, we examined the response of the mast cells and goblet cells over a period of 6 weeks in the duodenum, jejunum and ileum of C3H/HeN and C57BL/6 mice infected with Echinostoma hortense (E. hortense). In addition, we investigated whether the worm recovery rate of uninfected mice (the control group) or E. hortense-infected mice (the experimental group) was associated with the number of mast cells and goblet cells. The worm recovery rate was higher in the C3H/HeN mice than in the C57BL/6 mice. The number of goblet cells significantly increased in the experimental group of the C3H/HeN and C57BL/6 mice compared with the control group of both strains (P<0.005). Worm recovery peaked 3 weeks after the infection of the C57BL/6 mice and at 2 weeks after the infection of the C3H/HeN mice, and it was higher in the duodenum than in the jejunum and ileum. However, the infected site in the intestine had no relation with worm expulsion. In the C3H/HeN and C57BL/6 mice, the number of goblet cells in the experimental group was significantly higher than that in the control group (P<0.005). The number reached a peak 2 weeks after the infection and it even increased in duodenum, jejunum and ileum. The increased number of goblet cells was retained 6 weeks after infection. The number of goblet cells was higher in the C3H/HeN mice than in the C57BL/6 mice (P<0.01). These results indicate that goblet cells are related with the worm expulsion. Furthermore, immunohistostaining of the antral intestinal walls for lectin showed the significant increase of the number of goblet cells in the experimental group (P<0.001). The high infection rate in the duodenum was found during the early infection. An increased infection rate in the jejunum and ileum was found 3 weeks after infection and the infection rate was higher in the C3H/HeN mice than in the C57BL/6 mice. Taken together, the present study indicates that goblet cells, rather than mast cells, may play critical roles in parasite expulsion.

• The Plant Pathology Journal
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• v.18 no.5
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• pp.259-265
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• 2002

Apple mosaic virus (ApMV), a member of the genus Ilarvirus, was detected and isolated from diseased 'Fuji' apple (Malus domestica) in Korea. The coat protein (CP) genes of two ApMV strains, denoted as ApMV-Kl and ApMV-K2, were amplified by using the reverse transcription and polymerase chain reaction (RT-PCR) and were analyzed thereafter. The objectives were to define the molecular variability of genomic information of ApMV found in Korea and to develop virus-derived resistant gene source for making virus-resistant trans-genic apple. RT-PCR amplicons for the APMVS were cloned and their nucleotide sequences were determined. The CPs of ApMV-Kl and ApMV-K2 consisted of 222 and 232 amino acid residues, respectively. The identities of the CPs of the two Korean APMVS were 93.1% and 85.6% at the nucleotide and amino acid sequences, respectively. The CP of ApMV-Kl showed 46.1-100% and 43.2-100% identities to eight different ApMV strains at the nucleotide and amino acid levels, respectively. When ApMV-PV32 strain was not included in the analysis, ApMV strains shared over 83.0% and 78.6% homologies at the nucleotide and amino acid levels, respectively. ApMV strains showed heterogeneity in CP size and sequence variability. Most of the amino acid residue differences were located at the N-termini of the strains of ApMV, whereas, the middle regions and C-termini were remarkably conserved. The APMVS were 17.(1-54.5% identical with three other species of the genus Ilarviyus. ApMV strains can be classified into three subgroups (subgroups I, II, and III) based on the phylogenetic analysis of CP gene in both nucleotide and amino acid levels. Interestingly, all the strains of subgroup I were isolated from apple plants, while the strains of subgroups II and III were originated from peach, hop, or pear, The results suggest that ApMV strains co-evolved with their host plants, which may have resulted in the CP heterogeneity.

• Parasites, Hosts and Diseases
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• v.35 no.3
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• pp.189-196
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• 1997

The purpose of this study is to determine whether nitric oxide is involved in the extracellular killing of Trichomoncs uasinalis by mouse (BALB/c) peritoneal macrophages and RAW264.7 cells activated with LPS or rIFN-γ and also to observe the effects of various chemicals which affect the production of reactive nitrogen intermediates (RNl) in the cytotoxicity against T. vnginnlis. The cytotoxicity was measured by counting the release of (3H)-thymidine from labelled protozoa and NOa was assayed by Griess reaction. Nemonomethyl-L-arginine (L-NMHA), Nenitro-L-arginine methyl ester (NAME) and arginase inhibited cytotoxicity to T. vaginnlis and nitrite production by activated mouse perioneal macrophagrs and RAW 264.7 cells. The addition of excess L-arginine competitively restored trichomonacidal activity of macrophages. Exogenous addition of FeSO4 inhibited cytotoxicity to T. vaginaLis and nitric products of macrophages. From above results, it is assumed that nitric oxide plays an important role in the host defense mechanism of macrophages against T ucfinalis.

• Korean Journal of Veterinary Service
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• v.38 no.4
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• pp.233-239
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• 2015

Both Q-fever and Toxoplasmosis are zoonosis. Q-fever occurs due to intracellular bacteria, while Toxoplasmosis is created by protozoan. Both of them have a wide range of host including livestock and wild animals and occur sporadically all over in the world. In this study, seroprevalence of Q-fever and Toxoplasmosis was investigated on cows bred in the area of Seoul where there was a fairly high possibility to occur, while vaccine was not used in Korea. As for experiment materials, cattle blood collected from 190 cows from February to September in 2014 was used ELISA. According to the result, there was a positive reaction on Q-fever from 18 cows out of total 190 cows (9.5%) and on Toxoplasmosis from 32 cows (16.8%). Seroprevalence of both diseases per age was turned out to be negative for those aged less than 2. In addition, it was shown to be positive on 4 cows out of 87 (4.6%) cows aged from 3 to 5, on 7 cows out of 30 cows (23.3%) aged from 6 to 7. Finally, it was shown to be positive on 7 cows out of 17 cows (41.2%) aged 8 or above. Toxoplasmosis was turned out to be positive on 1 cow out of 56 cows (1.8%) aged less than 2, on 6 cows out of 87 cows (6.9%) aged from 3 to 5, on 17 cows out of 30 cows (56.7%) aged from 6 to 7. In addition, it was turned out to be 8 cows out of 17 cows (47.1%) aged 8 or above. Seroprevalence of both diseases was turned out to be higher as age increased. Therefore, it seems that a wide range of investigation on the entire disease spreaders as well as livestock is required since infection of Q-fever and Toxoplasmosis, types of zoonosis, has continuously occurred, and the number of insects, wild animals, and stray animals serving as a role of spreading diseases by changes in seasons and environments has been gradually increasing in Korea.

• Korean Journal of Veterinary Research
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• v.39 no.2
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• pp.247-256
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• 1999

A disease of young Holstein calves characterized by recurrent pneumonia, ulcerative and granulomatous stomatitis, enteritis with bacterial overgrowth, periodontitis, delayed wound healing, persistent neutrophilia and death at an early age had been originally described in 1983 and again in 1987. Most of these calves had stunted growth and a persistent, progressive neutrophilia (often exceeding 100,000/ml). By investigation of pedigrees, all of the affected calves have now been traced to a common sire and confirmed by polymerase chain reaction (PCR) diagnostic DNA testing to be homozygous carriers of a defective allele for bovine CD18. Neutrophils from these calves have several functional deficits and, most importantly, fail to adhere in a ${\beta}_2$-integrin dependent manner. The ${\beta}_2$-integrins represent a family of glycoproteins which participate in various leukocyte adhesion reactions during host defense. The presence or absence of ${\beta}_2$-integrin molecules can be demonstrated on the surface of neutrophils, monocytes and lymphocytes from normal or affected calves using specific monoclonal antibodies and flow cytometry, or by colloidal gold immunolabeling and scanning electron microscopy in backscatter mode. Deficiency of the ${\beta}_2$-integrins on all leukocyte types in Holstein calves is analogous to leukocyte adhesion deficiency (LAD) seen in humans. Neutrophils in bovine (BLAD) and human LAD patients are unable to adhere to the endothelial lining of the cardiovascular system thus interrupting egression of neutrophils into infected tissues. Other leukocytes, while still deficient in expression of the ${\beta}_2$-integrins, are still able to efficiently egress from the blood stream due to interactions of other adhesion molecules that are not as highly expressed on neutrophils. Both BLAD cattle and LAD children (who do not receive bone marrow transplants) often die at an early age as a result of the failure of neutrophils to extravasate into infected tissues. In 1991, Shuster, et $al^{27}$, identified two point mutations within the alleles encoding bovine CD18 in a Holstein calf afflicted with leukocyte adhesion deficiency. One mutation causes an aspartic acid to glycine substitution at amino acid 128 (D128G) in an extracellular region of this adhesion glycoprotein that is highly conserved (> 95% identity) between humans, cattle and mice. The other mutation is silent. Numerous calves with clinical symptoms of leukocyte adhesion deficiency have since been tested and all have been found homozygous for the D128G allele. In addition, calves homozygous far the D128G allele have been identified during widespread DNA testing in the United States. All cattle with the mutant allele are related to one bull, who through artificial insemination (A.I.), sired many calves in the 1950's and 1960's. The carrier frequency of the D128G CD18 allele among U.S. Holstein cattle had reached approximately 15% among active A.I. bulls and 8% among cows. By 1993, the organization of the dairy industry and the diagnostic test developed to genotype cattle, enabled virtually complete eradication of bovine leukocyte adhesion deficiency among current and future A.I. bulls.