• Title/Summary/Keyword: EAE

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Virulence Factor Profiles of Escherichia coli O157:H7 Bacteriophage Isolates from Sewage and Livestock Stools (하수와 가축분변에서 분리된 대장균 O157:H7 박테리오파지의 병원성인자 프로파일)

  • Seo, Jina;Seo, Dong Joo;Lee, Min Hwa;Jeon, Su Been;Oh, Hyejin;Oh, Mi Hwa;Choi, Changsun
    • Journal of Food Hygiene and Safety
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    • v.29 no.4
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    • pp.316-321
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    • 2014
  • The aim of study was to investigate the virulence profile of Escherichia coli O157:H7 bacteriophages isolated from sewage and livestock stools. Among 23 E. coli O157:H7 bacteriophages, 14 strains were isolated from sewage and 9 were from animal stools collected from 10 livestock farms in Korea. For each bacteriophage DNA sample, the presence of stx1, stx2, eae, aafII, ial, elt, estI, estII, astA, afa, and cnf was examined by polymerase chain reaction. The detection rate of eae, stx2, estI, astA, and ial was 100%, 69.6%, 13.0%, 13.0%, 8.7%, respectively. While all E. coli O157:H7 bacteriophages isolated from stools carried eae+stx2, stx2+eae, eae+astA, eae, stx2+eae+estI, eae+estI, stx2+eae+ial, and eae+ial were observed in bacteriophages isolated from sewage. As several plasmid-carrying virulence factors (estI, astA, and ial) were found in E. coli O157:H7 bacteriophages obtained from sewage and stools, the microbial safety of bacteriophages should be investigated in further study.

Recombination and Expression of eaeA Gene in Enterohemorrhagic Escherichia coli O157:H7

  • Kim, Hong;Kim, Jong-Bae
    • Biomedical Science Letters
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    • v.8 no.3
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    • pp.107-113
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    • 2002
  • Enterohemorrhagic Escherichia coli (EHEC) strains of serotype O157:H7 have been shown to colonize the intestinal epithelial cell by the attaching and effacing (AE) mechanism. The AE lesion is mediated by an intimin, of which production and expression are controlled by a 3-Kb eaeA gene located EHEC chromosomal DNA. If the eaeA gene is mutated, EHEC O157:H7 strains lose capacity of adhesion to intestinal epithelial cells. In this study, a 891 bp of the 3'-end region of a gamma intimin was amplified by polymerase chain reaction (PCR). The PCR product was inserted into pSTBlue-1 cloning vector and transformed into DE3 (BL21) competent cell. After plasmid mini-preparation and restriction enzyme digestion of eaeA/891-pSTBlue-1 vector, target eaeA gene was re-inserted into pET-28a expression vector and was transformed. Then the expression of recombinant eaeA/891 (891 bp) gene was induced by isopropyl-$\beta$-D-thiogalactopyranoside (IPTG). The expression of the 40-KDa recombinant protein was identified in SDS-PAGE and confirmed by immunoblotting using the His.Tag$^{\circledR}$ and T$_{7}$.Tag$^{\circledR}$ monoclonal antibody. This recombinant protein expressed by eaeA gene could be applied in further studies on the mechanisms of E. coli O157:H7 infection and the development of recombinant vaccine.

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Expression and localization of phospho-IkB in the central nervous system during experimental autoimmune encephalomyelitis in Lewis rats (자가면역성 뇌척수염 랫드의 중추신경계에서의 인산화된 IkB의 발현양상)

  • Hwang, In Sun;Jee, Young Heun
    • Korean Journal of Veterinary Research
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    • v.45 no.1
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    • pp.7-15
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    • 2005
  • To elucidate the roles of phospho-IkB expression in the development and progression of EAE, we investigated the expression of phospho-IkB in the central nervous system (CNS) of rats during experimental autoimmune encephalomyelitis (EAE) using immunohistochemistry and Western blot analysis. In Western blot analysis, the increased expression of phospho-IkB went parallel to severity of EAE. The expression of phospho-IkB increased significantly at the peak stage of EAE followed by gradual decrease. Immunohistochemical studies showed that the phospho-IkB immunoreactivity was mainly expressed in inflammatory cells (macrophages, T cells) and glial cells (astrocytes, microglial cells) at the peak stage of EAE and disappeared at the recovery stage. These findings suggest that the phosphorylation of IkB is closely associated with autoimmune inflammation in the CNS and plays an important role in the initiation and progression of EAE.

Chemical Stability and Whitening Activity of Mixture of Functional Ingredients (미백 기능성 성분의 혼합물에 대한 미백 활성 및 안정성에 관한 연구)

  • Lee, Youn-Hee;Lim, Nam-Hoon;Joung, Min-Seok;Kim, Joong-Hoi;Sim, Sang-Soo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.4 s.54
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    • pp.295-304
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    • 2005
  • Recently, arbutin, oil soluble licorice extact (GLY), ascorbyl glucoside (AA2G), and ethyl ascorbyl ether (EAE) have been widely used as functional whitening ingredients. To Investigate which combination between the above agents could be more effective for whitening effect, tyrosinase activity and MSH-induced melanin production in B-16 melanoma cells were investigated. Both GLY and arbutin dose-dependently inhibited purified tyrosinase activity. The inhibitory effects of GLY with AA2A or EAE on Drosinase activity were more potent than those of GLY alone, whereas that of arbutin with other ingreadients did not show those effects. In MSH-induced melanin production in B-16 melanoma cells, the mixture of Gly and EAE more significantly reduced melanin formation than Gly alone. Stability of mixture of GLY, arbutin, AA2A and EAE exposed at the temperature of $25^{\circ}C\;or\;45^{\circ}C$ for 30 days were also investigated. All of the combinations of whitening agents did not show any critical changes in their composition stability. These data suggest that the combination of GLY and vitamin C derivatives such as AA2G and EAE may be useful for the promotion of whitening effect.

Expression of protein kinase C in the spinal cords of rats with autoimmune encephalomyelitis (뇌염모델에서 Protein Kinase C의 발현에 관한 연구)

  • Shin, Tae-Kyun;Kim, Hyung-Min;Tanuma, Naoyuki;Matsumoto, Yoh
    • Korean Journal of Veterinary Pathology
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    • v.1 no.1
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    • pp.26-32
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    • 1997
  • Protein kinase C an enzyme of signal transduction has been known to regulate cell proliferation activation as well as apoptosis in some cancer cell lines. To explore the role of PKC in the course of cell mediated autoimmune disease such as experimental autoimmune encephalomyelitis (EAE) EAE was induced in Lewis rats(6-8 weeks old) with immunization of myelin basic protein supplemented with complete Freund's adjuvants and affected spinal cords were sampled at days 13 postimmunization(PI) as peak stage of EAE and at days 21 PI as recovery stage. The spinal cords with EAE were subjected to Northern blot analysis and insitu hybridization of PKC delta which is one of prominant isotypes of PKC in the haematopoietic cells. Northern blot analysis showed that levels of PKS delta mRNA in the spinal cords of rats withEAE was significantly increased at days 13 PI in which inflammatory cells including T cells and macrophages in the EAE lesions appeared. however the stage. By in situ hybridization signals of PKC delta in EAE lesions was intensely expressed on the delta is also expressed on some brain cells in normal rat central nervous system This finding suggests that PKC plays an important role on either activation of inflammatory cells including encephalitogenic T cells and macrophages or apoptotic elimination of some inflammatory cells depending on the stge of EAE.

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Histopathological evaluation of the lungs in experimental autoimmune encephalomyelitis

  • Sungmoo Hong;Jeongtae Kim;Kyungsook Jung;Meejung Ahn;Changjong Moon;Yoshihiro Nomura;Hiroshi Matsuda;Akane Tanaka;Hyohoon Jeong;Taekyun Shin
    • Journal of Veterinary Science
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    • v.25 no.3
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    • pp.35.1-35.13
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    • 2024
  • Importance: Experimental autoimmune encephalomyelitis (EAE) is an animal model of multiple sclerosis characterized by inflammation within the central nervous system. However, inflammation in non-neuronal tissues, including the lungs, has not been fully evaluated. Objective: This study evaluated the inflammatory response in lungs of EAE mice by immunohistochemistry and histochemistry. Methods: Eight adult C57BL/6 mice were injected with myelin oligodendrocyte glycoprotein35-55 to induce the EAE. Lungs and spinal cords were sampled from the experimental mice at the time of sacrifice and used for the western blotting, histochemistry, and immunohistochemistry. Results: Histopathological examination revealed inflammatory lesions in the lungs of EAE mice, characterized by infiltration of myeloperoxidase (MPO)- and galectin-3-positive cells, as determined by immunohistochemistry. Increased numbers of collagen fibers in the lungs of EAE mice were confirmed by histopathological analysis. Western blotting revealed significantly elevated level of osteopontin (OPN), cluster of differentiation 44 (CD44), MPO and galectin-3 in the lungs of EAE mice compared with normal controls (p < 0.05). Immunohistochemical analysis revealed both OPN and CD44 in ionized calcium-binding adapter molecule 1-positive macrophages within the lungs of EAE mice. Conclusions and Relevance: Taken together, these findings suggest that the increased OPN level in lungs of EAE mice led to inflammation; concurrent increases in proinflammatory factors (OPN and galectin-3) caused pulmonary impairment.

Change of blood pressure end vasorelaxation on EAE-induced lewis rat

  • Bong su Kang;Park, Young shim;In hoi Huh
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.194-194
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    • 1996
  • Pathogenesis of experimental allergic encephalomyelitis was involved with infection, inflammatory reaction, immune reaction etc. We studied on relation of blood vessel system and EAE. So, we measured blood pressure, heart rate and relaxation of isolated blood vessel in control and EAE-induced lewis rats. Blood pressure was decreased before EAE clinical sign (0-20day), but was increased from 23day. In isolated blood vessel, acetylcholine-treated relaxation was different on control, maximum EAE stage, recovery stage. Acetylcholine-treated relaxation was reduced 30% in recovery stage. but, sodium nitroprusside had similar relaxation reaction.

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ET-1 RIA and Immunocytochemistry on EAE-induced lewis rat

  • Bongsu Kang;Park, Youngshim;Inhoi Huh;Taekyun Shin
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.193-193
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    • 1996
  • Endothelin-1, which is a peptide originally isolated from cultured porcine aortic cell, has been found to play a crucial role in potentiating the vasoconstriction mitogenesis, and chemotaxis. In the present study, we examined the level of endothelin-1 in the brain, spinal cord and blood from rat with experimental allergic encephalomyelitis(EAE). At the peak stage of EAE(grade 3), endothelin-1 level in the spinal cord of rat with EAE increased two folds as compared with that of sham-treated rats, and subsequently decreased to the level of control at the recovery stage. In the endothelin-1 immunocytochemistry, endothelin-1 immunopositive cells and ED-1, macrophage marker immunopositive cells observed in the spinal cord of peak stage(grade 3). These Findings suggest that endothelin-1 play the important role in progression of EAE.

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BQ-123, $ET_{A}$ antagonist, decreases clinical sign and inflammatory region on EAE.

  • Kang, Bong-Su;Park, Young-Shim;In hoi Huh
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.191-191
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    • 1996
  • BQ-123, a selective $ET_{A}$ receptor antagonist, reverses various responses induced by Endothelin-1 and it has been reported that BQ-123 ameliorates the cerebrovascular constriction, hypertension, and decrease of blood flow. Previously, we announced that the level of Endothelin-2 increase in the brain and spinal cord of EAE-induced lewis rat and showed the origin of ET-1 is activated macrophages. Intracisternal injection of $ET_{A}$ receptor antagonist, BQ-123(10nmol) was done for visualizing the role of endothelin-1 on the pathogenesis of EAE. BQ-123 apparently blocked the severity of clinical score of EAE and decreased the histologically observed inflammatory region. The blocking effect on the progression of EAE model following BQ-123, suggests that BQ-123 is a physiological antagonist in terms of development of the sign of multiple sclerosis.

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Antimicrobial Resistance Profiles of eae Positive Escherichia coli (eae+ Escherichia coli의 항생제 감수성 및 내성 패턴)

  • Lee, Min-Hwa;Choi, Chang-Sun
    • Journal of Food Hygiene and Safety
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    • v.22 no.2
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    • pp.116-119
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    • 2007
  • The antimicrobial susceptibility and antibiotic resistance patterns of 67 eae positive Escherichia coli strains isolated from pigs were investigated by disc diffusion method. Sixty-seven E. coli isolated from pigs showed susceptibility to Ceftiofur (98.5%), Lincomycin+Spectinomycin (74.6%), Danofloxacin (73.1%), Enrofloxacin (64.2%), and Neomycin (41.8%). However, the multiple resistance patterns were also seen in eae+E. coli isolates. Neomycin+Tylosin+Penicillin+Tetracycline, Tylosin+Penicillin+Tetracycline, and Neomycin+Tylosin+Danofloxacin+Penicillin+Tetracycline+Enrofloxacinwere the most prevalent patterns of multiple antibiotic resistance.