• Title/Summary/Keyword: M-toxin

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Acid Resistance of Non-O157 Shiga Toxin-Producing Escherichia coli Adapted in Fruit Juices in Simulated Gastric Fluid (위합성용액에서 과일주스에 노출한 Non-O157 Shiga Toxin-Producing Escherichia coli의 산 저항성 평가)

  • Kim, Gwang-Hee;Oh, Deog-Hwan
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.4
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    • pp.577-584
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    • 2016
  • The objectives of this study were I) to compare the acid resistance (AR) of seven non-O157 Shiga toxin-producing Escherichia coli (STEC) serogroups, including O26, O45, O103, O111, O121, O145, and O157:H7 STEC isolated from various sources, in 400 mM acetic acid solution (AAS) at pH 3.2 and $30^{\circ}C$ for 25 min with or without glutamic acid and II) to determine strain survival upon exposure to simulated gastric fluid (SGF, pH 1.5) at $37^{\circ}C$ for 2 h after acid adaptation in apple, pineapple, orange, and strawberry juices at pH 3.8, $4^{\circ}C$ and $20^{\circ}C$. Results show that the O111 serogroup strains had the strongest AR (0.12 log reduction CFU/mL) which was very similar to that of O157:H7 STEC (P>0.05), compared to other serogroups in AAS without glutamic acid, whereas O26 serogroup strains showed the most sensitive AR. However, there was no significant (P>0.05) difference of AR among seven serogroups in AAS with glutamic acid. In the SGF study, 05-6545 (O45:H2), 08023 (O121:H19), and 03-4669 (O145:NM) strains adapted in fruit juices at $4^{\circ}C$ and $20^{\circ}C$ displayed enhanced survival with exposure to SGF for 60 min compared to 06E0218 (O157:H7) strains (P<0.05). In addition, 4 STEC strains adapted in pineapple juice at $4^{\circ}C$ showed enhanced survival with exposure to SGF for 60 min compared to those strains acid-adapted in the other fruit juices. Generally, adaptation at $4^{\circ}C$ in fruit juices resulted in significantly enhanced survival levels compared to acid-adapted at $20^{\circ}C$ and non-adapted conditions. The AR caused by adaptation in fruit juices at low temperature may thus increase survival of non-O157 STEC strain in acidic environments such as the gastrointestinal tract. These results suggest that more careful strategies should be provided to protect against risk of foodborne illness by non-O157 STEC.

A yeast Chromosomal Gene that Induces Defective Interfering Particles of L-A dsRNA Virus in $ski^-$ Host Cells ($ski^-$ 기주 세포에서 L-A dsRNA 바이러스의 defective interfering particle을 유도하는 효모 유전자)

  • 이현숙
    • Korean Journal of Microbiology
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    • v.29 no.2
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    • pp.75-79
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    • 1991
  • The yeast L-A virus (4.6 kb dsRNA genome) encodes the major coat protein and a "gag-pol" fusion minor coat protein that separately encapsidate itself and $M_{1}$, a 1.8 kb dsRNA satellite virus encoding a secreted protein toxin (the killer toxin). The teast chromosomal SKI genes prevent viral cytopathology by lowering the virus copy number. Thus, $ski^{-}$ mutants are ts and cs for growth. We transformed a ski2-2 virus-infested mutant with a yeast bank in a high copy cloning vector and selected the rare healthy transformants for analysis. One type of transformant segregated M-O L-A-O cells with high frequency. Elimination of the DNA clone from the ski2-2 strain eliminated this phinotype and introduction of the DNA clone recovered from such transformants into the parent ski2-2 strain, or into ski3 or ski6 mutants gave the same phenotype. This killer-curing phenotype was due to the curing of the helper L-A dsRNA virus. The 6.5 kb insert only had this activity when carried on a high copy vector and in $ski^{-}$ cells (not in $SKI^{+}$ cells). This 6.5 kb insert acts as a mutagen on L-A dsRNA producing a high rate of deletion mutations.mutations.

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Screening of Peptides Bound to Anthrax Protective Antigen by Phage Display

  • Kim, Joung-Mok;Park, Hye-Yeon;Choi, Kyoung-Jae;Jung, Hoe-Il;Han, Sung-Hwan;Lee, Jae-Seong;Park, Joon-Shik;Yoon, Moon-Young
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1784-1790
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    • 2006
  • Bacillus anthracis is a causative agent of anthrax. Anthrax toxins are composed of a protective antigen (PA), lethal factor (LF), and edema factor (EF), in which the PA is a central mediator for the delivery of the two enzymatic moieties LF and EF. Therefore, the PA has been an attractive target in the prevention and vaccinization for anthrax toxin. Recently, it has been reported that the molecule consisting of multiple copies of PA-binding peptide, covalently linked to a flexible polymer backbone, blocked intoxification of anthrax toxin in an animal model. In the present study, we have screened novel diverse peptides that bind to PA with a high affinity (picomolar range) from an M13 peptide display library and characterized the binding regions of the peptides. Our works provide a basis to develop novel potent inhibitors or diagnostic probes with a diverse polyvalence.

Optimized Culture Conditions for Production of the chimaeric protein, Uropathogenic Escherichia coli Adhesin - Cholera Toxin A2B Subunits, in Escherichia coli TB1

  • Lee, Yong-Hwa;Kim, Byung-Oh;Rhee, Dong-Kwon;Pyo, Suh-Kneung
    • Biomolecules & Therapeutics
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    • v.12 no.3
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    • pp.179-184
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    • 2004
  • The FimH subunit of type 1-fimbriated Escherichia coli has been determined as a major cause for urinary tract infections. In our previous study, the Adhesin/CTXA2B was expressed as soluble recombinant chimaeric protein derived from the uropathogenic Escherichia coli adhesin genetically coupled to cholera toxin A2B (CTXA2B) subunit in Escherichia coli. Since it is very important to optimize IPTG concentration and culture temperature to maximize cell growth and productivity, These optimal culture factors were determined to increase the productivity of the expressed Adhesin/CTXA2B chimaeric protein in Escherichia coli TB1 carrying pMALfimH/ctxa2b. Our data demonstrate that optimal concentration of IPTG for increased production of chimaeric protein was 0.5 mM. Additionally, culture time was 10 hours and temperature, 37${\circ}C$.

Prevalence of antibody and toxin against edema disease from pig farms in Jeonbuk province (전북지역 양돈장에서의 돼지 부종병 항체 및 톡신 양성률 조사)

  • Sun-Young Cho;Jeong Hee Yu;Yeong Ju Yu;Han-Jun Lee;Jin Hur
    • Korean Journal of Veterinary Service
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    • v.46 no.4
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    • pp.325-334
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    • 2023
  • Edema disease (ED) in pigs is enterotoxemia caused by Shiga toxin type 2e (Stx2e)-producing Escherichia coli (STEC) and frequently occurs in young piglets. Therefore, ED causes enormous economic losses in pig farms. In this study, a modified Stx2e (mStx2e) antigen was expressed and purified using commercial E. coli expression system. Monoclonal antibody was serviced by Ynto Ab Inc., using Phage Display Technique. Anti-Stx2e antibodies in piglets were measured by indirect ELISA using mStx2e antigens. Naive Stx2e in piglets were detected by Sandwich ELISA using Stx2e-monoclonal antibodies and commercial Stx2e-polyclonal antibodies. Among 3,480 piglets, anti-Stx2e antibodies were observed in 2,573 piglets. The 49.4% among 830 piglet serum samples possessed 0.625 ㎍/mL or more of Stx2e proteins. The 18.3% of 830 sera had 0.313 ㎍/mL of Stx2e proteins. The 32.3% of 830 samples held 0.156 ㎍/mL or less of Stx2e proteins. These results show that indirect ELISA using mStx2e antigen and Sandwich ELISA using Stx2e-monoclonal and polyclonal antibodies can be useful to detect ED in piglets.

The Toxin Purification and Isolation Identification of Meloidogyne hapla Toxicity Bacteria (Meloidogyne hapla 독성세균의 분리 동정 및 독성물질의 정제)

  • 이광배
    • Journal of environmental and Sanitary engineering
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    • v.14 no.2
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    • pp.32-39
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    • 1999
  • The following is experimental result of selecting soil bacteria showing toxicity against Root-knot nematode (Meloidogyne hapla). Out of 286 strains isolated from soil, one(NC67) showing toxicity against M.hapla is selected The selected strain(NC67) is identified of B. thuringiensis subsp. indiana. It proved out that the toxic maerial against M. hapla produce by NC67 strain is an exotoxin. The result of examining the existence of the extercellular toxicity product by the toxic strain(NC67) by usign activated carbon column chromatography, Dowex 50W column chromatography and TLC of silical gel etc. proved out that it is a single material.

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Modification of Insect Sodium Currents by a Pyrethroid Permethrin and Positive Cooperativity with Scorpion Toxins (피레스로이드계 살충제 퍼메트린이 Heliothis virescens 중추신경세포에 있는 나트륨채널에 작용하는 기작을 전기생리학적으로 연구)

  • Lee, Daewoo;Adams, Michael E.
    • Korean journal of applied entomology
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    • v.61 no.1
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    • pp.117-128
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    • 2022
  • In this study, we have examined pyrethroid actions on sodium channels in the pest insect Heliothis virescens. The synthetic pyrethroid permethrin increased steady-state sodium current in H. virescens central neurons and prolonged tail currents (INa-tail) due to extreme slowing of sodium channel deactivation. Prolongation of INa-tail was evident at permethrin concentrations as low as 60 nM, which modified ~1.7% of sodium channels and 10 μM permethrin modified about 30% of channels. The average time constant (τ1) of tail current decay was ~335 ms for permethrin-modified channels. These modified channels activated at more negative potentials and showed slower activation kinetics, and failed to inactivate. Permethrin modification of sodium channels was dramatically potentiated by the α scorpion toxin LqhαIT, showing positive cooperativity between two binding sites. The amplitude of the tail current induced by 0.3 μM permethrin was enhanced ~8-fold by LqhαIT (200 pM). Positive cooperativity was also observed between permethrin and the insect-specific scorpion toxin AaIT as 10 nM permethrin potentiated the shift of voltage dependence caused by AaIT (~2-fold).

Susceptibility of Anthonomus grandis (Cotton Boll Weevil) and Spodoptera frugiperda (Fall Armyworm) to a Cry1Ia-type Toxin from a Brazilian Bacillus thuringiensis Strain

  • Grossi-De-Sa, Maria Fatima;De Magalhaes, Mariana Quezado;Silva, Marilia Santos;Silva, Shirley Margareth.Buffon;Dias, Simoni Campos;Nakasu, Erich Yukio Tempel;Brunetta, Patricia Sanglard Felipe;Oliveira, Gustavo Ramos;De Oliveira Neto, Osmundo Brilhante;De Oliveira, Raquel Sampaio;Soares, Luis Henrique Barros;Ayub, Marco Antonio Zachia;Siqueira, Herbert Alvaro Abreu;Figueira, Edson L.Z.
    • BMB Reports
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    • v.40 no.5
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    • pp.773-782
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    • 2007
  • Different isolates of the soil bacterium Bacillus thuringiensis produce multiple crystal (Cry) proteins toxic to a variety of insects, nematodes and protozoans. These insecticidal Cry toxins are known to be active against specific insect orders, being harmless to mammals, birds, amphibians, and reptiles. Due to these characteristics, genes encoding several Cry toxins have been engineered in order to be expressed by a variety of crop plants to control insectpests. The cotton boll weevil, Anthonomus grandis, and the fall armyworm, Spodoptera frugiperda, are the major economically devastating pests of cotton crop in Brazil, causing severe losses, mainly due to their endophytic habit, which results in damages to the cotton boll and floral bud structures. A cry1Ia-type gene, designated cry1Ia12, was isolated and cloned from the Bt S811 strain. Nucleotide sequencing of the cry1Ia12 gene revealed an open reading frame of 2160 bp, encoding a protein of 719 amino acid residues in length, with a predicted molecular mass of 81 kDa. The amino acid sequence of Cry1Ia12 is 99% identical to the known Cry1Ia proteins and differs from them only in one or two amino acid residues positioned along the three domains involved in the insecticidal activity of the toxin. The recombinant Cry1Ia12 protein, corresponding to the cry1Ia12 gene expressed in Escherichia coli cells, showed moderate toxicity towards first instar larvae of both cotton boll weevil and fall armyworm. The highest concentration of the recombinant Cry1Ia12 tested to achieve the maximum toxicities against cotton boll weevil larvae and fall armyworm larvae were 230 ${\mu}g/mL$ and 5 ${\mu}g/mL$, respectively. The herein demonstrated insecticidal activity of the recombinant Cry1Ia12 toxin against cotton boll weevil and fall armyworm larvae opens promising perspectives for the genetic engineering of cotton crop resistant to both these devastating pests in Brazil.

Evaluation of Oxidative DNA Damage Using an Alkaline Single Cell Gel Electrophoresis (SCGE) Comet Assay, and the Protective Effects of N-Acetylcysteine Amide on Zearalenone-induced Cytotoxicity in Chang Liver Cells

  • Kang, Changgeun;Lee, Hyungkyoung;Yoo, Yong-San;Hah, Do-Yun;Kim, Chung Hui;Kim, Euikyung;Kim, Jong Shu
    • Toxicological Research
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    • v.29 no.1
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    • pp.43-52
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    • 2013
  • Zearalenone (ZEN) is a non-steroidal estrogenic mycotoxin produced by several species of Fusarium that are found in cereals and agricultural products. ZEN has been implicated in mycotoxicosis in farm animals and in humans. The toxic effects of ZEN are well known, but the ability of an alkaline Comet assay to assess ZEN-induced oxidative DNA damage in Chang liver cells has not been established. The first aim of this study was to evaluate the Comet assay for the determination of cytotoxicity and extent of DNA damage induced by ZEN toxin, and the second aim was to investigate the ability of N-acetylcysteine amide (NACA) to protect cells from ZEN-induced toxicity. In the Comet assay, DNA damage was assessed by quantifying the tail extent moment (TEM; arbitrary unit) and tail length (TL; arbitrary unit), which are used as indicators of DNA strand breaks in SCGE. The cytotoxic effects of ZEN in Chang liver cells were mediated by inhibition of cell proliferation and induction of oxidative DNA damage. Increasing the concentration of ZEN increased the extent of DNA damage. The extent of DNA migration, and percentage of cells with tails were significantly increased in a concentration-dependent manner following treatment with ZEN toxin (p < 0.05). Treatment with a low concentration of ZEN toxin (25 ${\mu}M$) induced a relatively low level of DNA damage, compared to treatment of cells with a high concentration of ZEN toxin (250 ${\mu}M$). Oxidative DNA damage appeared to be a key determinant of ZEN-induced toxicity in Chang liver cells. Significant reductions in cytolethality and oxidative DNA damage were observed when cells were pretreated with NACA prior to exposure to any concentration of ZEN. Our data suggest that ZEN induces DNA damage in Chang liver cells, and that the antioxidant activity of NACA may contribute to the reduction of ZEN-induced DNA damage and cytotoxicity via elimination of oxidative stress.

Application of Botulinum Toxin on Masticatory Muscle of Patients with Bruxism

  • Jang, Seok-Min;Jeon, Hye-Mi;Kim, Kyung-Hee;Ok, Soo-Min;Heo, Jun-Young;Jeong, Sung-Hee;Ahn, Yong-Woo
    • Journal of Oral Medicine and Pain
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    • v.39 no.2
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    • pp.55-62
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    • 2014
  • Purpose: This study aims to evaluate the changes in soft tissue thickness of the masseteric region after injection of botulinum toxin type A (BTX-A). Methods: Twenty-four data acquired from medical records were classified into 4 groups: bruxer group that received masseter muscle injection only (M-B), bruxer group that received both masseter and temporalis muscle injections (MT-B), non-bruxer group that received masseter muscle injection only (M-NB) and non-bruxer group that received both masseter and temporalis muscle injections (MT-NB). Injection dose of BTX-A was 30 units for each masseter muscle and 20 units for each temporalis muscle. We measured the reduced thickness of the masseteric region before and after 12 weeks after injection using cone-beam computed tomography. Results: Among the patients that received both masseter and temporalis muscle injections, bruxer group showed a tendency to have more reduction in masseter muscle thickness than non-bruxer group. The difference in reduced thickness between M-B and MT-B tended to show greater than the difference between M-NB and MT-NB. Conclusions: In case of masseter hypertrothy patients with bruxism there was a tendency to show a difference in reduced thickness of soft tissue between the group that received both masseter and temporalis muscles injection and the group that received masseter muscle injection only hence a thorough inspection before the injection of BTX-A is condisered to be needed.