• 대한수의학회지
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• 제36권2호
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• pp.379-387
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• 1996

The objects of the present study were to establish the method of purification, subunit dissociation of verotoxin-2 (VT2) produced by Escherichia coli O157:H7, and to investigate the characteristics of purified verotoxin-2 such as molecular weight and composition of amino acid. The results were summerized as follows; Verotoxin-2 was extracted by addition of polymyxin B sulfate into bacterial cell lysate prepared from Escherichia coli O157:H7(KSC109). As an initial step, the bacterial cell lysate was precipitated with 30% saturated ammonium sulfate. The precipitated crude toxin was then subjected to anion-exchange, chromatofocusing and cation-exchange chromatography. Using this scheme, we obtained highly purified toxin with a specific activity of $1.1{\times}10^9$ $CD_{50}/mg$. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) for purified VT2 showed two protein bands. The upper band, approximately 32 Kd, was supposed as A subunit and the lower band, approximately 7.7 Kd, was supposed as B subunit. When the toxin was separated in the subunit-dissociating solution, two peaks emerged with retention times of 15 and 28 min by HPLC. These peaks represented A subunit and B subunit, respectively. The amino acid composition of purified VT2 were made up in order of glutamic acid, histamine, asparaginic acid, histidine, lysine, alanine and leucine etc. The largest amount among the amino acid composing VT2 was methionine.

• The Plant Pathology Journal
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• 제35권5호
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• pp.445-458
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• 2019

The lipopolysaccharide (LPS) composed of lipid A, core, and O-antigen is the fundamental constituent of the outer membrane in gram-negative bacteria. This study was conducted to investigate the roles of LPS in Burkholderia glumae, the phytopathogen causing bacterial panicle blight and seedling rot in rice. To study the roles of the core oligosaccharide (OS) and the O-antigen region, mutant strains targeting the waaC and the wbiFGHI genes were generated. The LPS profile was greatly affected by disruption of the waaC gene and slight reductions were observed in the O-antigen region following wbiFGHI deletions. The results indicated that disruption in the core OS biosynthesis-related gene, waaC, was associated with increased sensitivity to environmental stress conditions including acidic, osmotic, saline, and detergent stress, and to polymyxin B. Moreover, significant impairment in the swimming and swarming motility and attenuation of bacterial virulence to rice were also observed in the waaC-defective mutant. The motility and virulence of O-antigen mutants defective in any gene of the wbiFGHI operon, were not significantly different from the wild-type except in slight decrease in swimming and swarming motility with wbiH deletion. Altogether, the results of present study indicated that the LPS, particularly the core OS region, is required for tolerance to environmental stress and full virulence in B. glumae. To our knowledge, this is the first functional study of LPS in a plant pathogenic Burkholderia sp. and presents a step forward toward full understanding of B. glumae pathogenesis.

• 대한수의학회지
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• 제41권4호
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• pp.511-521
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• 2001

Microorganisms were isolated and identified from bovine 296 quarters which showed positive reaction by California Mastitis Test (CMT) in 40 farms of Jeju from September 1999 to June 2000. The organisms associated with the mastitis of bovine were 11 different bacterial species in this study. Which of them, Staphylococcus aureus was the most predominant species as 152 (51.4%) isolates. Other identified species included 49 (16.5%) coliform, 47 (15.8%) Streptococcus dysgalactiae, 15 (5.1%) Bacillus spp., 8 (2.7%) Staphylococcus epidermidis, 6 (2.1%) Streptococcus agalactiae, 5 (1.7%) Enterococcus faecalis, 5 (1.7%) Corynebacterium spp., 3 (1.0%) Streptococcus uberis, 1 (0.3%) Pseudomonas aeruginosa and 1 (0.3%) Pasteurella haemolytica. Almost of all the islolated beacterial species showed high sensitivity against kanamycin (98.6%), cephalothin (98.0%), streptomycin (94.9%), gentamicin (94.6%), ampicillin (92.2%) and polymyxin B (90.2%). On the contrary, they showed resistance against penicillin (47.0%), tetracycline (37.2%), cefazolin (26.0%), bacitracin (22.6%) and erythromycin (19.9%). Eighty-one isolates were not resistant to any antibiotics and 215 drug resistant isolates showed 89 different drug resistance patterns from single to nine multiple antibiotics resistance patterns.

• Biomolecules & Therapeutics
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• 제19권3호
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• pp.302-307
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• 2011

Peptidoglycan (PGN) is detected in inflammatory cell-rich regions of human atheromatous plaques. The present study investigated the effects of PGN on CC chemokine ligand 3 (CCL3) expression, which is elevated in the atherosclerotic arteries, and determined cellular factors involved in PGN-mediated CCL3 up-regulation in mononuclear cells, with the goal of understanding the molecular mechanisms of inflammatory responses to bacterial pathogen-associated molecular patterns in diseased arteries. Exposure of human monocytic leukemia THP-1 cells to PGN resulted in enhanced secretion of CCL3 and profound induction of the CCL3 gene transcript. Both events were abrogated by oxidized 1-palmitoyl-2-arachidonosyl-sn-phosphatidylcholine, an inhibitor of Toll-like receptors 2/4. Pharmacological inhibitors such as U0126, SP6001250, Akt inhibitor IV, rapamycin, RO318220, diphenyleneiodonium chloride, and N-acetylcysteine also significantly attenuated PGN-mediated CCL3 up-regulation. However, polymyxin B, LY294002, and SB202190 did not influence CCL3 expression. We propose that PGN contributes to enhanced CCL3 expression in atherosclerotic plaques and that Toll-like receptors (TLR2), Akt, mTOR, mitogen-activated protein kinase, and reactive oxygen species are involved in that process.

• 한국임상수의학회지
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• 제19권2호
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• pp.159-164
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• 2002

The present study was conducted to investigate the antibiotic resistance of 24 Salmonella gallinarum isolated from 48 chicken samples of diagnosed fowl typhoid cases during the period from November 1998 to November 1999. And the isolates of S gallinarum were also tested for their invasion abilities to experimental infection as one of virulence tests, and the presence of virulence-related plasmid in S gallinarum isolates. The results obtained through this study were summarized as follows; 1. All of isolates from 48 cases of 24 farms were identified S gallinarum by biochemical and serological tests.2. Antimicrobial drug resistance test against 24 isolates showed that the isolates were resistant to Colistin(95.8%), and Penicillin(79.2%), Polymyxin B(75.0%), Streptomycin (65.2%), Gentamycin(54.2%), and Tetracycline(33.3%). 3. Mortality in chicken following peroral inoculation of four isolates of S gallinarum during 14-days inoculation pecked at 5 days(40%) after inoculation and all of experimental chickens died within 13 days after inoculation.4. Based on the pattern and number of isolated plasmid from each isolate, plasmid profiles were divided into five groups; group I with 3 plasmids, group II to group IV with 4 plasmids and group V with 5 plasmids.

• 한국동물위생학회지
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• 제22권2호
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• pp.121-128
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• 1999

The present study was conducted to investigate the incidence of pneumonic lesions with special regard to enzootic pneumonia and the microbiology of pneumoic lungs from 544 slaughter pigs during the period from October 1995 to September 1996. The incidence of enzootic pneumonic lesion was 76.3% (41s/s44) and pleurisy was detected from 7.9% of slaughter pigs. Seasonal prevalence of pneumonic lesions in slaughter pigs were in order of prevalence of 82.9% in spring, 76.8% in winter, 74.8% in autumn and 69.0% in summer, respectively. Pasteurella multocida, Streptococcus sp, Str suis, Corynebacterium sp, Actinobacillus pleuropneumoniae, Hemophilus parasuis, and Klebsiella pneumoniae were detected in order of prevalence from 16.9%, 15.9%, 7.5%, 6.0%, 1.4%, 1.0% and 0.5% of 415 pneumonic lungs, respectively. P multocida were susceptible to oxytetracycline, polymyxin-B, streptomycin, and vancomycin, while the majority of them were resistant to amoxicillin, ampicillin, cephalothin, kanamycin, and penicillin-G. Str suis were susceptible to amoxicillin, ampicillin, cephalothin, penicillin-G, although the majority of them were resistant to erythromycin, oxytetracycline, streptomycin, vancomycin. A pleuropneumoniae were susceptible to ampicillin, and cephalothin, but the majority of them were resistant to oxytetracycline.

• International Journal of Oral Biology
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• 제38권2호
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• pp.73-80
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• 2013

Leptin is one of the adipocytokines produced from adipose tissue but its functions in periodontal tissue have not previously been investigated. In our current study, we examined the effects of leptin on the expression of interleukin (IL)-6 and IL-8 in periodontal ligament (PDL) cells and gingival fibroblasts. Leptin receptor expression was evaluated by RT-PCR and the production of cytokines was measured by ELISA. The phosphorylation of Akt and Erk1/2 was assessed by western blotting. mRNA of long and short form leptin receptors were detected in both PDL cells and gingival fibroblasts. Leptin was found to increase the production of IL-6 and IL-8 in both of these cell types, an effect which was not blocked by polymyxin B, an inhibitor of lipopolysaccharide (LPS). Leptin did not alter the production of IL-6 and IL-8 induced by LPS in PDL cells but increased Akt and Erk1/2 phosphorylation in these cells. These results suggest that leptin acts as an inducer of IL-6 and IL-8 in PDL cells and gingival fibroblasts.

• 대한피부과학회지
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• 제56권10호
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• pp.624-627
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• 2018

Allergic contact dermatitis is an inflammatory condition associated with periorbital erythema, edema, and pruritus. The periorbital skin is relatively thin compared with the skin over other facial areas; therefore, it is vulnerable to allergen penetration and may show a variety of cutaneous manifestations. Recently, vision enhancement surgery is a widely performed procedure, and the prevalence of senile cataract and glaucoma is increasing. The prevalence of periocular allergic contact dermatitis is increasing secondary to the growing use of topical ophthalmic medications. Several studies in Korea have reported periocular allergic contact dermatitis secondary to the use of topical ophthalmic medications including latanoprost ($Latano^{(R)}$), fluorometholone ($Tolon^{(R)}$), polymyxin B ($Terramycin^{(R)}$), atropine sulfate ($Atropine^{(R)}$), neomycin sulfate ($Cambison^{(R)}$), and befunolol hydrochloride ($Bentos^{(R)}$), among others. However, ofloxacin ($Effexin^{(R)})$-induced allergic contact dermatitis has not been reported in the domestic and/or foreign literature. We report a case of periocular allergic contact dermatitis secondary to the use of ofloxacin ophthalmic ointment.

• Journal of the Korean Wood Science and Technology
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• 제20권4호
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• pp.73-84
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• 1992

It was observed that the hot-water extract of the bark of Kalopanax pictus N. had the highest anti-complementary activity among the 11 kinds of forest materials. Methanol-and ethanol-soluble portions had low anti-complementary activities, but crude polysaccharide. HKP-0 had a high activity of 80%. HKP-0 contained 54.8% of total sugar and 27.9% of protein. The neutral sugars of HKP-0 consisted of mainly arabinose, galactose and glucose. HKP-4 fraction obtained by cetavlon treatment of HKP-0 showed the highest anti-complementary activity of 90%. The activity was not changed by pronase digestion bu decreased greatly by periodate oxidation. HKP-4 consisted of mainly arabinose and glucose with molar ratio of 1.0 : 22.4, HKP-4-I, an unabsorbed fraction from HKP-4 on DEAE Sepharose CL-6B column showed higher yield and activity than those of absorbed fractions. HKP-4-I was homogeneous, and its molecular weight was about 25,000. HKP-4-I contained 84.0% of neutral sugar and consisted of arabinose and glucose with molar ratio of 1.0 : 11.2. The anti-complementary activity of HKP-4-I was not decreased by the treatment of polymyxin B, and the polysaccharide activated both classical and alternative pathway in complement system. Void volume fraction obtained from HKP-4-I hydrolyzed with ${\alpha}$-amylase on Sephadex G-25 column only had a high anti-complementary activity.

• 생명과학회지
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• 제16권1호
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• pp.131-135
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• 2006

미생물이 이용할 수 있는 nitrogen source는 di-, tri-, oli- go-peptide 또는 amino acid의 형태로 세포내로 uptake되어 대사과정에 사용되고 있다. 이와같은 peptide는 특이한 transport system에 의해서 이동되고 있는데 oligo peptide(Opp) transport system에는 binding protein, permease protein, energy 생성을 위한 ATP 분해에 관여하는 protein 이 관여하고 있으며 염색체 상에서 이들 단백질들은 operon 형태의 유전자로부터 발현되고 있다. 본 연구는 gram 음성 세균이며 수해양 서식 세균인 V, fluvialis로부터 얻어진 Opp operon 유전자 가운데 oligopeptide binding protein을 coding하고 있는 oppA 유전자가 deletion된 mutant를 사용하여 여러 환경변화에 따른 생육을 wild type과 비교한 연구 결과 이다. 생육을 위한 완전배지인 brain heart infusion (BHI) 배지와 최소배지인 M9 minimal 배지를 사용한 결과 OppA protein의 생성 결핍에 따라 초기 및 대수증식기 과정 중에는 mutant의 생육이 늦어지고 있으나 Opp system이 아닌 다른 peptide전달 경로로 추정되는 system을 이용하여 대수 증식기 후반에서는 wild type과 거의 같은 생육 형태를 보여 주고 있었다. pH의 변화에 따른 생육은 pH 7에서는 생육정도가 비슷하였으나 약알칼리 부근에서는 oppA mutant의 생육이 wild type에 비하여 낮아지고 있었다. 또한 5 mM $H_2O_2$를 사용하여 $OD_{600}=1.2$농도의 세포들에 대한 영향을 검토한 결과 두 균 모두 높은 생존율을 보여 주었으며 이는 대수증식기 세포들을 사용한 결과와는 매우 다른 형태를 보여 주고 있었다. 항생제 내성에 대한 연구에서는 mutant가 streptomycin과 tetracycline 에 대해서는 wild type과는 다르게 매우 낮은 농도에서도 생육이 되고 있지 않으나 polymyxin B에 대해서는 wild type과 같이 $10{\mu}g/ml$의 농도에서도 잘 자라고 있었다.