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Experimental In Vivo Models of Bacterial Shiga Toxin-Associated Hemolytic Uremic Syndrome

  • Jeong, Yu-Jin;Park, Sung-Kyun;Yoon, Sung-Jin;Park, Young-Jun;Lee, Moo-Seung
    • Journal of Microbiology and Biotechnology
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    • v.28 no.9
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    • pp.1413-1425
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    • 2018
  • Shiga toxins (Stxs) are the main virulence factors expressed by the pathogenic Stx-producing bacteria, namely, Shigella dysenteriae serotype 1 and certain Escherichia coli strains. These bacteria cause widespread outbreaks of bloody diarrhea (hemorrhagic colitis) that in severe cases can progress to life-threatening systemic complications, including hemolytic uremic syndrome (HUS) characterized by the acute onset of microangiopathic hemolytic anemia and kidney dysfunction. Shiga toxicosis has a distinct pathogenesis and animal models of Stx-associated HUS have allowed us to investigate this. Since these models will also be useful for developing effective countermeasures to Stx-associated HUS, it is important to have clinically relevant animal models of this disease. Multiple studies over the last few decades have shown that mice injected with purified Stxs develop some of the pathophysiological features seen in HUS patients infected with the Stx-producing bacteria. These features are also efficiently recapitulated in a non-human primate model (baboons). In addition, rats, calves, chicks, piglets, and rabbits have been used as models to study symptoms of HUS that are characteristic of each animal. These models have been very useful for testing hypotheses about how Stx induces HUS and its neurological sequelae. In this review, we describe in detail the current knowledge about the most well-studied in vivo models of Stx-induced HUS; namely, those in mice, piglets, non-human primates, and rabbits. The aim of this review is to show how each human clinical outcome-mimicking animal model can serve as an experimental tool to promote our understanding of Stx-induced pathogenesis.

Development of Automated Algorithm for Compartment Arrangement of Oil Tanker (유조선의 구획배치 자동화 알고리즘 개발)

  • Song, Ha-Cheol;Na, Seung-Su;Jo, Du-Yeon;Shim, Cheon-Sik;Lee, Gang-Hyeon;Jeong, Sol;Heo, Joo-Ho;Jeong, Tae-Seok;Lee, Chul-Ho;Jo, Young-Chun;Kim, Dong-Chun
    • Journal of the Society of Naval Architects of Korea
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    • v.50 no.1
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    • pp.59-68
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    • 2013
  • Nowadays, optimum structural design techniques based on CSR have been developed and applied to the preliminary design stage focused on minimum weight and minimum construction cost of ship structure. Optimum structural design algorithm developed before could minimize weight and cost on fixed compartment arrangement. However, to develop more efficient design technique, a designer needs to combine optimized compartment arrangement with optimized ship structural design because compartment arrangement has a large effect on structural design according to the change of still water bending moment as a consequence of compartment arrangement change. In this study, automated algorithm for compartment arrangement of an oil tanker is developed to apply preliminary structural design. The usefulness of developed algorithm is verified with Aframax oil tanker constructed by STX shipbuilding Co.Ltd..

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.

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.

Experimental Study on Sloshing Loads in a B-Type Independent Tank (독립형 화물창 내부의 슬로싱 하중에 대한 실험적 연구)

  • Kim, Sang-Yeob;Ahn, Yangjun;Kim, Yonghwan;Heo, Joo-Ho;Jeong, Taeseok;Lee, Chul-Ho;Kim, Do-Hyun
    • Journal of the Society of Naval Architects of Korea
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    • v.50 no.6
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    • pp.390-398
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    • 2013
  • In this paper, an experimental study on sloshing problems in an independent B-type tank of STX Offshore and Shipbuilding Co. is described. Recently STX Offshore and Shipbuilding Co. introduced a new design of an independent B-type tank in order to reduce sloshing impact loads on LNG CCS. This tank has many internal members, so that sloshing flow and the resultant hydrodynamic loads are very different from those in typical membrane tanks. In this study, a series of sloshing experiment have been carried out for 1/50 scale model, and the main characteristics of sloshing load on the independent tank are observed. The properly scaled internal members such as swash bulkhead, center bulkhead and stringers have been installed in the test tank model, but sloshing pressures are measured on the tank walls only. The forced excitation signals have been generated by using the predicted ship motion in irregular sea states. The characteristics of sloshing loads on this tank have been observed in different filling levels with various heading angles, and sea states. In this paper, some key findings from the model tests are discussed.

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.

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.

Paralytic Shellfish Poison Profile in Commercial Shellfishes (시판 중인 패류의 마비성 패류독 특성)

  • Jang, Jun-Ho;Yun, So-Mi;Lee, Jong-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.6
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    • pp.924-928
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    • 2005
  • Toxin profiles of the paralytic shellfish poison (PSP) detected from domestic shellfishes collected at the market and imported. shellfishes were invested by fluorometric HPLC. Total 9 components in PSP were analysed from the imported ark shell, such as saxitoxin (STX), decarbamoylsaxitoxin (dcSTX), gonyautoxin (GTX) - 1,2,3,4,5, Cl and C2. Among those toxins, 7 components except for GTX1,4 were detected from domestic shellfishes and showed different toxin contents and toxin compositions by species. Only C group toxin (Cl +2) contained in short necked clam and hard clam $(0.06\~0.56\;nmole/g)$ which living under soil but, in the blue mussels and oysters which cultured in the open sea water, showed more higher toxicity and complicate toxin compositions. Toxin compositions in bloody clam and purplish washington clam were very different in some samples even in same species. GTX4 and GTX5 were higher in imported scallop and STX was higher in imported ark shell than other species.

Paralytic Shellfish Poisoning Toxin Accumulation in Four Mussel Species Fed on Toxic Alexandrium tamarense (독성 Alexandrium tamarense를 섭취한 담치류 4종의 마비성패독 축적)

  • Kim, Young-Soo;Shon, Myung-Back;Kim, Chang-Hoon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.39 no.1
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    • pp.49-54
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    • 2006
  • Cultured cells of the toxic Alexandrium tamarense were fed to four mussel species, Mytilus coruscus, M. edulis, M. galloprovincialis and Septifer vulgatus, to examine the interspecies and interlocality differences in the ability to accumulate paralytic shellfish poisoning (PSP) toxins. Toxin content of A. tamarense cells varied during culture period. In contrast, toxin composition in the cell (C1,2, GTX1-4 and neoSTX) was constantly stable. In feeding experiment, the four mussel species collected from Geoje intoxicated after uptake of A. tamarense. Toxin content ($average{\pm}SD\;{\mu}g$ STXeq/100 g) of M. coruscus, M. edulis, M. galloprovincialis and Septifer vulgatus were $1,660{\pm}79,\;3,914{\pm}2,242,\;5,626{\pm}1,620\;and\;958{\pm}163$, respectively. Toxin profiles included C1,2, GTX1,4 and neoSTX as the major components, and dcGTX2,3, GTX2,3, neoSTX and STX as the minor ones. Toxin accumulation of three mussel species collected from Pohang, Geoje and Anmyon-do showed interspecies and interlocality differences. Toxin content ($average{\pm}SD\;{\mu}g$ STXeq/100 g) were $91{\pm}4,\;151{\pm}14,\;39{\pm}3$ in M coruscus, $189{\pm}1,\;231{\pm}11,\;206{\pm}15$ in M edu/is and $214{\pm}28,\;326{\pm}30,\;291{\pm}26$ in M. galloprovincialis in order of Anmyon-do, Geoje and Pohang.

The Study of Full Penetration Welding between Corrugated BHD and Lower Stool Joint by Application of CSR (CSR 적용에 따른 Corrugated BHD와 Lower Stool Joint의 Full Penetration Welding 적용에 관한 연구)

  • Park, Chan-Kyu;Yang, Jong-Soo;Kim, Ho-Kyung
    • Special Issue of the Society of Naval Architects of Korea
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    • 2007.09a
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    • pp.135-141
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    • 2007
  • CSR(Common Structure Rules) enter into force on $1^{st}$ April 2006. Generally for double hull tankers of less than 150m in length, the Rules of the individual Classification Society are to be applied. Where high tensile stresses act through an intermediate plate, increased fillet welds or penetration welds are to be used longitudinal/transverse bulkhead primary support member end connections to the double bottom. If workers have begun to make used of established procedures between corrugated BHD and lower stool joint, first to welding on groove of face and then it has to gouging to blow on groove of root. So amount of man-hour increased, productivity secreased.

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