• Title/Summary/Keyword: stx2

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Identification and characterization of Shiga toxin-producing Escherichia coli isolated from diarrhea in calves (송아지 설사분변으로부터 Shiga toxin-producing Escherichia coli 의 분리 및 특성규명)

  • Lim, Keum-Gi;Kang, Mun-Il;Kim, Snag-Ki;Nam, Kyung-Woo;Park, Hyun-Joo;Park, Jin- Ryang;Cho, Kyoung-Oh;Lee, Bong-Joo
    • Korean Journal of Veterinary Research
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    • v.46 no.2
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    • pp.135-142
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    • 2006
  • Shiga toxin (stx) producing Escherichia coli (STEC) causes various clinical signs in animal and human. In this study, 255 fecal samples from calves showing diarrhea were collected from cattle farms in Chonnam province during the period from January 2005 to July 2005. Twenty six STEC (10%) were isolated from 255 fecal samples by PCR. The isolates displayed three different stx combinations (stx1 [69%], stx1 and stx2 [15%], and stx2 [38%]). The isolates were further studied for virulence associated genes and antimicrobial resistance to define the virulence properties. Intimin (eaeA), enterohemolysin (hlyA), and lipopolysaccharide (rfbE) virulence genes were detected in 6 (23%), 7 (26%), and 1 (3.8%) of the isolates, respectively, by PCR. One isolate possessing rfbE gene was typed as E. coli 0157 : H7 by agglutination test with O and H antisera. All 26 isolates showed susceptibility to amikacin (100%) and the majority of isolates showed high susceptibility to gentamicin (88.5%) and chloramphenicol (73.1%). But all isolates were resistant to penicillin. These results may provide the basic knowledge to establish strategies for the treatment and prevention of enteric disease in calves.

Induction of Deletion Mutation for the Enzymatic Domain in the Shigatoxin2e A Subunit Gene of Esherichila coli O139 Isolates and Expression of Mutated Protein (분리 대장균 O139의 Shigatoxin2e A 유전자의 효소 활성부에 대한 결손변이 유발 및 변이 단백질의 발현)

  • Cho Eun-jung;Kim Do-kyong;Kim Sang-hyun;Kim Yeong-il;Lee Chul-hyun;Lee Woo-won;Son Won-geun;Shin Jong-Uk;Kim Yong-hwan
    • Journal of Veterinary Clinics
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    • v.22 no.4
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    • pp.386-391
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    • 2005
  • This study was done to produce a mutated protein inactivated cytotoxicity of Shigatoxin 2e (Stx2e) of E.coli O139 isolates by deletional mutagenesis of Stx2e A subunit gene encoding active-site cleft of enzymatic domain in ST2e holotoxin. Cytotoxicity of the toxoid expressed from the mutant Stx2e gene was compared with wild type Stx2e for development of vaccine candidate. A recombinant plasmid pED18 containing Stx2e gene ot E.coli O139 isolates was used to generate mutation plasmid. Deletion mutagenesis was conducted for Stx2e A subunit gene encoding enzymatically active domain by polymerase chain reaction (PCR) using ot designed primer to induce deletional mutation. DNA sequence analysis was confirmed that the pentamer (Typ 202- Ser 206) that lies within the proposed active-site cleft in the second region was completely deleted. A DNA fragment of 1.1 kb that encode the new mutant Stx2eA gene was inserted into plasmid pRSET vector digested with EcoRV-Hind III and named pEDSET The PEDSET was transformed in E. coli for expression of mutant protein and the protein was confirmed by SDS-PACE and Western-blotting. The protein expressed by the mutant was tested to confirm the reduction of cytotoxic activities on Vero cell using microcytotoxicity assay compared with wild type Stx2e, the cytotoxicity of deletional mutant protein was at least reduced by 3,000-fold on Vero cell.

Consideration of Structural Damage of Fresh Water Tank for 2,600 TEU Container Vessel (2,600 TEU Container Vessel 의 Fresh Water Tank 구조손상 사례 고찰)

  • Shin Sung-Kwang;Ahn Hyung-Joon;Choi Eui-keol;Koh Myeong-Seob;Leem Hyo-Kwan
    • Special Issue of the Society of Naval Architects of Korea
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    • 2005.06a
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    • pp.216-221
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    • 2005
  • Many tanks such as a fresh water tank, an aft peak tank and oil tanks are arranged in the engine room and aft part areas of the ship. By added mass effect of the fluid inside the tanks, the natural frequency will be changed according to filling height of the tank. For this reason, there is possibility of occurrence of excessive vibration by resonance between natural frequencies of local structure and excitation frequencies of the propeller or main engine. Therefore, calculation of natural frequencies is required for structure for many types of tank which are contacting with water or oil to consider added mass effect for anti-resonance design at design stage. In this study, a case of structure damage on the fresh water tank for 2600 TEU container vessel is introduced. In addition, natural frequency analysis and vibration measurement have been performed to investigate vibration characteristics for excessive vibration control.

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Assessment of Fatigue Crack Propagation Considering the Redistribution of Residual Stress due to Overload

  • Jang, Chang-Doo;Leem, Hyo-Kwan;Choi, Yeoung-Dal;Bang, Jun-Kee;So, Ha-Young
    • Journal of Ship and Ocean Technology
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    • v.11 no.2
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    • pp.26-33
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    • 2007
  • For the assessment of the retardation of fatigue crack propagation behavior due to overload, new FE analysis algorithms considering compressive residual stress redistribution near crack tip was proposed in this paper. The size of plastic zone near crack tip was obtained by elasto-plastic analysis and it was compared with Irwin's equation. The amount of residual stress redistribution was assessed by subsequent elasto-plastic analysis, and the difference of residual stress distributions between constant amplitude load and overload was obtained. In the analysis of fatigue crack propagation, the applied SIF range was evaluated by ASTM E647, and the effect of residual stresses on crack propagation was considered using the effective SIF concept. The test results of crack propagations were compared with the predicted data obtained by the analysis.

Consideration of second intact stability criteria (2세대 비손상 복원성 기준에 대한 고찰)

  • Park, Kwang-soo;Park, Han-chae;Lee, Chul-ho
    • Special Issue of the Society of Naval Architects of Korea
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    • 2017.10a
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    • pp.19-27
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    • 2017
  • In spite of adoption of Intact Stability Code Part A as mandatory, the accidents have been occurred frequently caused by dynamic stability. Therefore International Maritime Organization (IMO) is revising the Intact Stability Code to prevent the cause of accidents. The result of 4th meeting of Sub-committee on Ship Design and Construction (SDC), five mode of the cause was defined and establishing each draft regulations. We consider physical background, mathematical description, draft regulations for five failure mode and sample calculations for ships built by STX Offshore & Shipbuilding.

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Comparison of MBA and HPLC Post-column Oxidation Methods for the Quantification of Paralytic Shellfish Poisoning Toxins

  • Yu, Hongsik;Lim, Keun Sik;Song, Ki Cheol;Lee, Ka Jeong;Lee, Mi Ae;Kim, Ji Hoe
    • Fisheries and Aquatic Sciences
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    • v.16 no.3
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    • pp.159-164
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    • 2013
  • The mouse bioassay and high performance liquid chromatography (HPLC) post-column oxidation method are different methods of quantifying paralytic shellfish poisoning toxins. In this study, we compared their ability to accurately quantify the toxicity levels in two types of field sample (oysters and mussels) with different toxin profiles for routine regulatory monitoring. A total of 72 samples were analyzed by both methods, 44 of which gave negative results, with readings under the limit of detection of the mouse bioassay ($40{\mu}g/100g$ saxitoxin [STX] eq). In 14 oysters, the major toxin components were gonyautoxin (GTX) 1, -2, -3, -4, -5, decarbamoylgonyautoxin-2 (dcGTX2), and decarbamoylsaxitoxin (dcSTX), while 14 mussels tested positive for dcSTX, GTX2, -3, -4, -5, dcGTX2, neosaxitoxin (NEO), STX, and dcSTX. When the results obtained by both methods were compared in two matrices, a better correlation ($r^2=0.9478$) was obtained for mussels than for oysters ($r^2=0.8244$). Additional studies are therefore needed in oysters to investigate the differences in the results obtained by both methods. Importantly, some samples with toxin levels around the legal limit gave inconsistent results using HPLC-based techniques, which could have a strong economic impact due to enforced harvest area closure. It should therefore be determined if all paralytic shellfish poisoning toxins can be quantified accurately by HPLC, and if the uncertainties of the method lead to doubts regarding regulatory limits.

Shiga Toxins Trigger the Secretion of Lysyl-tRNA Synthetase to Enhance Proinflammatory Responses

  • Lee, Moo-Seung;Kwon, Haenaem;Nguyen, Loi T.;Lee, Eun-Young;Lee, Chan Yong;Choi, Sang Ho;Kim, Myung Hee
    • Journal of Microbiology and Biotechnology
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    • v.26 no.2
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    • pp.432-439
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    • 2016
  • Shiga toxins (Stxs) produced by Shiga toxin-producing Escherichia coli (STEC) strains are major virulence factors that cause fatal systemic complications, such as hemolytic uremic syndrome and disruption of the central nervous system. Although numerous studies report proinflammatory responses to Stx type 1 (Stx1) or Stx type 2 (Stx2) both in vivo and in vitro, none have examined dynamic immune regulation involving cytokines and/or unknown inflammatory mediators during intoxication. Here, we showed that enzymatically active Stxs trigger the dissociation of lysyl-tRNA synthetase (KRS) from the multi-aminoacyl-tRNA synthetase complex in human macrophage-like differentiated THP-1 cells and its subsequent secretion. The secreted KRS acted to increase the production of proinflammatory cytokines and chemokines. Thus, KRS may be one of the key factors that mediate transduction of inflammatory signals in the STEC-infected host.

Study on Convergence Technique Using the Antimicrobial Resistance and Virulence Genes Analysis in Escherichia coli (대장균의 항균제 내성과 독력 유전자의 분석을 활용한 융합기술연구)

  • Han, Jae-Il;Sung, Hyun-Ho;Park, Chang-Eun
    • Journal of the Korea Convergence Society
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    • v.6 no.5
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    • pp.77-84
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    • 2015
  • This study was conducted to investigate the characteristics of antibiotic resistant E. coli. its antibiotic susceptibility and pathogenicity were analyzed via molecular convergence technique, for the relationship of antibiotic susceptibility and pathogenicity. The 60 isolated strains consisted of ESBL(+)(8) and ESBL(-)(52) strains. The ESBL(+)(8) strains consisted of 2 strains without a pathogenic gene, stb(3), flich7(1), and flich7-eae(2). The ESBL(-)(52) strains consisted of 26 strains without a pathogenic gene, stx1(3), stb(10), flich7(2), eae(2), stx1-flich7(2), stx1-stb(4), flich7-stb(2), and flich7-stb-eae(1). In conclusion, antibiotic resistance is increasingly, Focused on molecular convergence, showed the correlation of pathogenicity with antibiotic resistance was poor. However, It will be able to find the exact pathogenic factor in the future through convergence technique including the analysis of virulence genes.

Efficiency of PetrifilmTM Staph Express Count Plate for the Enumeration of Staphylococcus aureus in Meat, Fishery Product, and Korean Traditional Foods (축·수산·전통식품 중 황색포도상 구균의 정량적 분석을 위한 PetrifilmTM Staph Express Count Plate의 성능 평가)

  • Yoo, Yoonjeong;Choi, Yuna;Choi, Seungho;Bang, Hyunjo;Yoon, Yohan;Ha, Jimyeong;Lee, Soomin
    • Journal of Food Hygiene and Safety
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    • v.34 no.3
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    • pp.309-315
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    • 2019
  • To enumerate Staphylococcus aureus in food, Baird-Parker Agar (BPA) is usually used in the conventional method, However it requires time and space for the preparation and plating, and incubation. Thus, use of the $3M^{TM}$ $Petrifilm^{TM}$ Staph Express Count Plate (STX Petrifilm) might be appropriate to solve these challenging problems. The purpose of this study was to compare the efficiency of STX Petrifilm with BPA for enumeration of S. aureus in various foods. A mixture of S. aureus strains ATCC29213, ATCC25923, and ATCC13565 was inoculated on marinated pork chop, beef (chuck tender), dried filefish, semi-dried squid, rice cake, and Japchae (stir-fried glass noodles) at 2, 3, 5, and 7 Log CFU/g. S. aureus cell counts were enumerated by spread-plating on STX Petrifilm and BPA after 0 and 24 hours at $4^{\circ}C$ (marinated pork chop, beef, semi-dried squid, and stir-fried glass noodles) and $25^{\circ}C$ (dried filefish and rice cake). Recovery of STX Petrifilm for S. aureus from various food samples was compared with BPA, and the results showed that there were no significant differences between two selective media in all cases. The results indicated that STX Petrifilm had enough efficiency to recover S. aureus from various foods as well as saving time and space.

Isolation and Purification of Lipopolysaccharide Derived from Escherichia coli O157:H7 for the Specific Antibody Production (병원성 Escherichia coli O157:H7의 특이 항체 생산을 위한 Lipopolysaccharide분리 및 정제)

  • 최학선;신영민;정숙현;박영민;안원근
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.3
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    • pp.571-575
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    • 2004
  • Escherichia coli O157:H7 cause hemorrhagic colitis and the extraintestinal complication of hemolytic-uremic syndrome, with their higher incidence occurring in children. Lipopolysaccharide (LPS) of E. coli O157:H7 is very important to make IgG anti-LPS with bactericidal activity. To identify the characteristic of E. coli OI57:H7, we isolated 60 MDa plasmid and amplified stx genes of shiga-like toxin (Stx) 1, 2 of E. coli O157:H7 by polymerase chain reaction (PCR) method. Using the simple purification method which contained phenol extract, ethanol precipitation and gel filtration steps, the LPS of E. coli O157:H7 was isolated and purified. Finally, we confirmed the purity of LPS through SDS-PAGE and silver nitrate staining.