• Title/Summary/Keyword: 1-2 toxin

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Characterization and isolation of shiga toxin-producing Escherichia coli from Bovine feces and Carcass (소의 분변과 도체에서 shiga toxin-producing Escherichia coli의 분리와 특성)

  • Chae, Hee-Sun;Kim, Neung-Hee;Han, Hye-Jin;Son, Hong-Rak;Kim, Chang-Ki;Kim, Sun-Heung;Lee, Jung-Hark;Kim, Jong-Taek
    • Korean Journal of Veterinary Service
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    • v.32 no.3
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    • pp.241-249
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    • 2009
  • Shiga toxin (Stx)-producing Escherichia coli (STEC) strains can cause broad spectrum of human disease, including diarrhea, hemorrhagic colitis, and the life-threatening hemolytic uremic colitis (HUS). We examined 868 samples was taken from bovine feces and carcass from January to December 2008 in Seoul. Twenty two (9.5%) shiga toxin -producing Escherichia coli were isolated from the 230 of bovine feces, and two (0.31%) were isolated from the 638 of carcasses. Serotype of E. coli isolates were O157 (10, 41.6%), O26 (10, 41.6%), O111 (1, 4.2%) and UT (3, 12.6%). In PCR, the isolates displayed three different stx gene combination (stx1 [2, 8.4%]), stx2 [3, 12.6%] and stx1 and stx2 [19,87.5%]). The eaeA and hlyA gene were found in 11 (45.8%) of the 24 strains. Saa gene was present only one strains (4.2%). Toxin typing using reverse passive latex agglutination test showed the same result in VT 1. But it showed different result in VT 2. In antimicrobial susceptibility test, all isolates were sensitive to amikacin, amoxicillin/clavulanic acid, ciprofloxacin and colistin. Eighteen strains (75.0%) of 24 isolates showed the multi-resistant patterns with over 3 drugs. PFGE was performed after the genomic DNA of twenty four isolates was digested with Xba I. the 24 isolates showed 7 (A~G) PFGE type.

Insecticidal Toxin from Xenorhabdus nematopilus, Sysbiotic Bacterium Associated with Entomopathogenic Nematode Sreinernema glaseri

  • Ryu, Keun-Garp;Bae, Jun-Sang;Yu, Yeon-Su;Park, Sun-Ho
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.2
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    • pp.141-145
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    • 2000
  • Entomopathogenic nematodes are being used for insect control. We purified a toxin secreted by the insect-pathogenic bacterium, Xenorhadbus nematophilus, which lives in the gut of entomopathogenic nematodes. Culture broth of X. nematophilus was separated by centrifugation and concentrated by ultration. The concentrated culture broth was applied to a DEAE Sephadex A-50 column, and proteins were eluted stepwise with increasing concentrations of KCI. Fractions column. The molecty weight of purified toxin was39 kDa on SDS-PAGE, and Fourier tranformed infrared (FTIR) spectroscopy indicated that this toxin could be a new protein exhiting the charactristics of C=O stretching peak near 1650cm-1.

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Increase of Spacer Sequence Yields Higher Dimer $(Fab-Spacer-Toxin)_{2}$ Formation

  • Yoo Mee-Hyeon;Won Jae-Seon;Lee Yong-Chan;Choe Mu-Hyeon
    • Journal of Microbiology and Biotechnology
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    • v.16 no.7
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    • pp.1097-1103
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    • 2006
  • The divalent antibody-toxins are expected to have increased binding avidities to target cells because of the two cell-binding domains. However, previous studies showed that the refolding yield of divalent antibody-toxin is very low, and it is assumed that homodimer formation of antibody-toxin is strongly interfered by the repulsion between the two large toxin domains that come close to each other during dimer formation. In this study, B3 antibody was used as a model antibody, and its Fab domain was used to construct three different kinds of Fab divalent molecules, $[B3(Fab)-toxin]_{2}$. The monomer Fab-toxin molecules were made by fusing the Fab domain of monoclonal antibody B3 to PE38, a truncated mutant form of Pseudomonas exotoxin (PE), and a connecting sequence that contained spacer amino acid sequence (G4S)n (n=l, 2, 3) was inserted between Fab and PE38. The prepared divalent molecules were $[Fab-S\;1,\;2,\;3-PE38]_{2}\;(=[Fab-SKPCIST-KAS(G_{4}S)nGGPE-PE38]_{2}\;(n=1,\;2,\;3))$, and they are derivatives of previously studied $[Fab-H2cys-PE38]_{2}\;(=[Fab-SKPCIST-KASGGPE-PE38]_{2})$. In $[Fab-Sl,\;2,\;3-PE38]_{2}$, two Fab-S1, 2, 3-PE38 monomers were covalently linked by the disulfide bond bridge made from cysteine in the -SKPCIST- sequence. The insertion of spacer amino acids after the disulfide bridge resulted in a 12-18 fold higher yield of dimer formation than previously constructed $[Fab-Hlcys-PZ38]_{2}[7]$, 3-4-fold higher than $[Fab-ext-PZ38]_{2}[25]$. These two molecules have less amino acid spacer sequence between the disulfide bridge and PE38 domain. The design of $[Fab-PE38]_{2}$ in this study gave molecules with a higher refolding yield. The results of cytotoxicity assay showed a higher cytotoxic effect of these divalent molecules than that of the monovalent scFv-PE38 molecule.

Antibiotic Resistance and Virulence Potentials of Shiga Toxin-Producing Escherichia coli Isolates from Raw Meats of Slaughterhouses and Retail Markets in Korea

  • Park, Hyun-jung;Yoon, Jang Won;Heo, Eun-Jeong;Ko, Eun-Kyoung;Kim, Ki-Yeon;Kim, Young-Jo;Yoon, Hyang-Jin;Wee, Sung-Hwan;Park, Yong Ho;Moon, Jin San
    • Journal of Microbiology and Biotechnology
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    • v.25 no.9
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    • pp.1460-1466
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    • 2015
  • In this study, the prevalence of Shiga toxin-producing Escherichia coli (STEC) was investigated among raw meat or meat products from slaughterhouses and retail markets in South Korea, and their potential for antibiotic resistance and virulence was further analyzed. A total of 912 raw meats, including beef, pork, and chicken, were collected from 2008 to 2009. E. coli strains were frequently isolated in chicken meats (176/233, 75.9%), beef (102/217, 42.3%), and pork (109/235, 39.2%). Putative STEC isolates were further categorized, based on the presence or absence of the Shiga toxin (stx) genes, followed by standard O-serotyping. Polymerase chain reaction assays were used to detect the previously defined virulence genes in STEC, including Shiga toxins 1 and Shiga toxin 2 (stx1 and 2), enterohemolysin (ehxA), intimin (eaeA), STEC autoagglutination adhesion (saa), and subtilase cytotoxin (subAB). All carried both stx1 and eae genes, but none of them had the stx2, saa, or subAB genes. Six (50.0%) STEC isolates possessed the ehxA gene, which is known to be encoded by the 60-megadalton virulence plasmid. Our antibiogram profiling demonstrated that some STEC strains, particularly pork and chicken isolates, displayed a multiple drug-resistance phenotype. RPLA analysis revealed that all the stx1-positive STEC isolates produced Stx1 only at the undetectable level. Altogether, these results imply that the locus of enterocyte and effacement (LEE)-positive strains STEC are predominant among raw meats or meat products from slaughterhouses or retail markets in Korea.

Effectiveness of Botulinum Toxin A in Treatment of Frey's Syndrome (Frey 증후군의 치료에 있어서 보툴리눔 독소 A형의 유용성)

  • Kim, Kook Hyun;Park, Eun Soo;Youn, Chang Won;Lee, Young Mann;Kim, Yong Bae
    • Archives of Craniofacial Surgery
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    • v.10 no.2
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    • pp.114-119
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    • 2009
  • Purpose: Several therapeutic approaches have been introduced and tried to treat Frey syndrome following parotidectomy. However they were not proved as an effective treatment. A new therapeutic modality using botulinum toxin injections was presented previously by several study groups. But, the duration of the demonstrated positive effect was essentially unknown so far. The purpose of this clinical investigation is to demonstrate the effectiveness of Botulinum toxin type A(BTXA) in patients with Frey syndrome. Methods: For this study, 12 patients were treated, They were assessed with the Minor's iodine-starch test and interviewed before and after treatment. Botulinum toxin is injected with $1.0cm^2$ apart into the skin where symptoms of Frey's syndrome has manifestated. The patients were classified according to the concentration of botulinum toxin and dosage of botulinum toxin. Results: The outcome measures were the time of reappearance of gustatory sweating(subjective study), and the results of an Minor's iodine-starch test(objective study) of 3 weeks, 3 months, 6 months, 9 months, 12 months after treatment. This treatment was effective in all groups of patients. The concentrations and the amount of dosages did not affect the treatment. But high concentration produced faster effectiveness in subjective. Conclusion: Botulinum toxin type A(BTXA) for Frey syndrome is easy, convenient and effective withouts severe complication. And the patients has no difficulty and limitation in their life.

Snake Venom synergized Cytotoxic Effect of Natural Killer Cells on NCI H358 Human Lung Cancer Cell Growth through Induction of Apoptosis

  • Oh, Jae Woo;Song, Ho Sueb
    • Journal of Acupuncture Research
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    • v.33 no.2
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    • pp.1-9
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    • 2016
  • Objectives : I investigated whether snake venom can synergistically strengthen the cytotoxic effects of NK-92 cells, and enhance the inhibition of the growth of lung cancer cells including NCI-H358 through the induction of death receptor dependent extrinsic apoptosis. Methods : Snake venom toxin inhibited cell growth of NCI-H358 Cells and exerted non influence on NK-92 cell viability. Moreover, when they were co-cultured with NK cells and concomitantly treated with $4{\mu}g/m{\ell}$ of snake venom toxin, more influence was exerted on the inhibition of growth of NCI-H358 cells than BV or NK cell co-culture alone. Results : The expression of Fas, TNFR2 and DR3 and in NCI-H358 lung cancer cells was significantly increased by co-culture of NK-92 cells and treatment of $4{\mu}g/m{\ell}$ of snake venom toxin, compared to co-culture of NK-92 cells alone. Coincidentally, Bax, caspase-3 and caspase-8 - expressions of pro-apoptotic proteins in the extrinsic apoptosis pathway, demonstrated significant increase. However, in anti-apoptotic NF-${\kappa}B$ activities, activity of the signal molecule was significantly decreased by co-culture of NK-92 cells and treatment of $4{\mu}g/m{\ell}$ of snake venom toxin, compared to co-culture of NK-92 cells or snake venom toxin treated by NCIH358 alone. Meanwhile, in terms of NO generation, there is a significant increase, in co-culture of NK-92 cells with NCI-H358 cells as well as the co-culture of NK-92 cells and concomitant treatment of $4{\mu}g/m{\ell}$ of snake venom toxin. However, no synergistic increase of NO generation was shown in co-culture of NK-92 cells and treatment of $4{\mu}g/m{\ell}$ of snake venom toxin, compared to co-culture of NK-92 cells with NCI-H358 cells. Conclusion : Consequently, this data provides that snake venom toxin could be useful candidate compounds to suppress lung cancer growth along with the cytotoxic effect of NK-92 cells through extrinsic apoptosis.

A Pilot Study to Evaluate the Efficacy and Safety of Treatment with Botulinum Toxin in Patients with Recalcitrant and Persistent Erythematotelangiectatic Rosacea

  • Park, Kui Young;Kwon, Hyun Jung;Kim, Jae Min;Jeong, Guk Jin;Kim, Beom Joon;Seo, Seong Jun;Kim, Myeong Nam
    • Annals of dermatology
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    • v.30 no.6
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    • pp.688-693
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    • 2018
  • Background: There are few pharmacologic options to reduce erythema and flushing in patients with recalcitrant erythematotelangiectatic rosacea (ETR). We previously reported two cases of refractory flushing and erythema of rosacea that were successfully treated with intradermal botulinum toxin injection, and additional research is needed to prove the efficacy and safety of this treatment. Objective: To report the efficacy and safety of botulinum toxin injection as an aid in persistent erythema of rosacea patients. Methods: A total of 20 Korean patients with recalcitrant ETR were enrolled to receive treatment by injection of botulinum toxin. Patients received one treatment of intradermal botulinum toxin injection and were assessed 1, 2, 4, and 8 weeks after treatment. The severity of erythema and telangiectasia was investigated by a non-treating physician, and the Erythema Index (EI) was assessed by mexameter at each visit. Patient satisfaction and any adverse events were also assessed at each visit. Results: 17 patients completed all follow-up visits and were included in the analysis. Intradermal injection of botulinum toxin significantly reduced erythema severity and EI in ETR patients. Patients reported a satisfaction score of $2.94{\pm}0.56$ at 8 weeks after treatment. Except for three patients who discontinued the study early due to inconvenience of facial muscle paralysis, 17 patients participating in the final analysis did not report side effects except injection pain at the time of the procedure. Conclusion: Intradermal injection of botulinum toxin can be used as an effective and relatively safe adjuvant agent for recalcitrant and persistent erythema of ETR patients.

Toxin Produced by Colletotrichum falcatum Causing Red Rot of Sugarcane

  • Saikia, R.;Azad, P.;Arora, D.K.
    • Mycobiology
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    • v.32 no.4
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    • pp.149-154
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    • 2004
  • Toxin produced by Colletotrichum falcatum Went, the incitant of red rot of sugarcane was isolated, purified and assayed to determine host specificity and identify its chemical nature. The toxin was found to be not host specific as it inhibited germination of various seeds(gram, greengram, blackgram, pea, cowpea, rice and sugarcane) as well as different seedlings viz. tomato, coriander, pea and rice. The toxin consists of two distinct fraction-one fraction having $R_f$, value at 0.36 producing identical red rot lesion when inoculated at leaf midrib of sugarcane, and the other having $R_f$, value at 0.72 not showing any red rot lesion. Chromatogram of high performance liquid chromatography(HPLC) of the red rot lesion causing fraction showed a sharp peak at 1.62 min of retention time(RT), and spectral analysis indicated the presence of following chemical $CH_3$ - groups-C-H, C=O, C-N, $-CH_3,\;-CH_2$ -CH and molecular mass of the compound was 203. - ($M^+,\;C_{11}H_{11}N_2O_2$).

Production of Host-specific Toxin by Alternaria mali and its Biological Activity (사과점무늬낙엽병균(Alternaria mali)이 생성(生成)하는 기주특이적(寄主特異的) 독소(毒素)와 그의 생물활성(生物活性))

  • Yu, Seung-Hun;Shim, Hyeong-Kwon;Park, Jong-Seong
    • Korean journal of applied entomology
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    • v.26 no.3 s.72
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    • pp.171-178
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    • 1987
  • Pathogenic isolates of Alternaria mali produced host-specific toxin(AM-toxin) in liquid culture. The toxin was also released by germinating spores of the fungus. The physiological event of apple leaves induced by germinating spores was an increased loss of electrolytes from susceptible leaves. This reaction was evident soon after spore inoculation, indicating that the leakage was caused by AM-toxin from germinating spores. Typical symptoms were developed only in susceptible leaves of apple within 48hr after inoculation with pathogenic spores. Similar symptoms occurred on susceptible leaves when non-pathogenic isolates plus AM-toxin were used.

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Biotoxic Cyanobacterial Metabolites Exhibiting Pesticidal and Mosquito Larvicidal Activities

  • Kumar, Ashok;Dhananjaya P. , Singh;Tyagi, M.B.
    • Journal of Microbiology and Biotechnology
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    • v.13 no.1
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    • pp.50-56
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    • 2003
  • A freshwater bloom-forming cyanobacterium, Microcystis aeruginosa, and local soil isolate Scytonema sp. strain BT 23 were demonstrated to contain biotoxic secondary metabolites with pesticidal and mosquito larvicidal activities. A purified toxic constituent from M aeruginosa showed an absorption maximum at 230 nm and its toxicity symptoms, Rf value on TLC, and retention time observed ill an HPLC analysis were similar to those of the hepatotoxic heptapeptide microcystin-LR. The bioactive constituent of the Scytonema sp. was less polar in nature and exhibited two peaks at 240 and 285 m. When applied to two cruciffrous pests, Pieris brassicae and Plutella flostella, the crude extracts and toxic principles from the two cyanobacteria showed significant antifeedant activity in a no-choice bioassay, and at higher concenuations exhibited contact toxicity to the insect larvae. The purified toxin from M. aeruginosa was found to be more effective and produced 97.5 and $92.8\%$ larval mortality in the two pests, fo11owing 2 h of toxin treatment at a concentration of $25{\mu}g$ Per leaf disc (2.5 cm dia.). Meanwhile, similar treatment with the purified toxin from Sytonema sp. stain BT 23 only produced 73 and $78\%$ mortality in the two pests. The cyanobacterial constituents also showed significant activity against Culex and Anopheles larvae. The M. aeruginosa toxin ($20{\mu}g\;ml^-1$) caused 98.2 and $88.1\%$ mortality in the Culex and Anopheles larvae, respectively, while the purified toxin from the Sytonema sp. was less toxic and only produced a 96.3 and $91.2\%$ mortality, respectively, at a much higher concentration ($40{\mu}g\;ml^-1$). Accordingly, the current results point to certain hitherto unknown biological properties of cyanobacterial biotoxins.