• Title/Summary/Keyword: toxin production

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Crotamine stimulates phagocytic activity by inducing nitric oxide and TNF-α via p38 and NFκ-B signaling in RAW 264.7 macrophages

  • Lee, Kyung Jin;Kim, Yun Kyu;Krupa, Martin;Nguyen, Anh Ngoc;Do, Bich Hang;Chung, Boram;Vu, Thi Thu Trang;Kim, Song Cheol;Choe, Han
    • BMB Reports
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    • v.49 no.3
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    • pp.185-190
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    • 2016
  • Crotamine is a peptide toxin found in the venom of the rattlesnake Crotalus durissus terrificus and has antiproliferative, antimicrobial, and antifungal activities. Herein, we show that crotamine dose-dependently induced macrophage phagocytic and cytostatic activity by the induction of nitric oxide (NO) and tumor necrosis factor-alpha (TNF-α). Moreover, the crotamineinduced expression of iNOS and TNF-α is mediated through the phosphorylation of p38 and the NF-κB signaling cascade in macrophages. Notably, pretreatment with SB203580 (a p38-specific inhibitor) or BAY 11-7082 (an NF-κB inhibitor) inhibited crotamine-induced NO production and macrophage phagocytic and cytotoxic activity. Our results show for the first time that crotamine stimulates macrophage phagocytic and cytostatic activity by induction of NO and TNF-α via the p38 and NF-κB signaling pathways and suggest that crotamine may be a useful therapeutic agent for the treatment of inflammatory disease.

Application of Functional Microbial Strains Isolated from Traditional Rice Wine in Korea (막걸리 유래 미생물의 활용을 위한 연구 동향)

  • Lee, Youngsuk;Seol, Jeongman;Jeong, Deokyeol;Kim, Soo Rin
    • Microbiology and Biotechnology Letters
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    • v.44 no.3
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    • pp.229-235
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    • 2016
  • Korea has a long history of brewing traditional rice wine using a authentic starter culture called nuruk, which contains natural microbial flora. Because rice wine is consumed fresh without filtration, its viable cells contribute to the biological activities of the wine. In numerous studies, microbial strains isolated from rice wine have been screened for their functionalities, which were mainly probiotic properties and antimicrobial activities. Indeed, some lactic acid bacteria (LAB) were confirmed to have strong probiotic activities as well as other health-promoting effects. Moreover, some of the isolated probiotic strains produced functional compounds, such as exopolysaccharides and γ-aminobutyric acid. For antimicrobial activities, some LAB and yeast strains were identified to produce bacteriocins and killer toxins, respectively, with significantly broad spectrum of antimicrobial activity. These functional strains originating from traditional rice wine and their metabolites can be used directly for the production of value-added food products.

Depression of $Ca^{2+}$ Influx in Complement C5a-stimulated Neutrophils by Calmodulin Inhibitors

  • Ham, Dong-Suk;Kim, Hyun-Ho;Han, Eun-Sook;Lee, Chung-Soo
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.1
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    • pp.109-117
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    • 1998
  • Role of $Ca^{2+}$/calmodulin complex in intracellular $Ca^{2+}$ mobilization in neutrophils has not been clearly elucidated. In this study, effects of chlorpromazine, trifluoperazine and imipramine on the intracellular $Ca^{2+}$ mobilization, including $Ca^{2+}$ influx, in C5a-activated neutrophils were investigated. Complement C5a- stimulated superoxide production and myeloperoxidase release in neutrophils were inhibited by chlorpromazine, trifluoperazine and imipramine, except no effect of imipramine on myeloperoxidase release. A C5a-elicited elevation of [$Ca^{2+}$]i in neutrophils was inhibited by chlopromazine, trifluoperazine, imipramine, staurosporine, genistein, EGTA, and verapamil but not affected by pertussis toxin. The intracellular $Ca^{2+}$ release in C5a-activated neutrophils was not affected by chlorpromazine and imipramine. Chlorpromazine and imipramine inhibited $Mn^{2+}$ influx by C5a-activated neutrophils. Thapsigargin-evoked $Ca^{2+}$ entry was inhibited by chlorpromazine, trifluoperazine, imipramine, genistein, EGTA and verapamil, while the effect of staurosporine was not detected. The results suggest that $Ca^{2+}$/calmodulin complex is involved in the activation process of neutrophils. The depressive action of calmodulin inhibitors on the elevation of cytosolic $Ca^{2+}$ level in C5a-activated neutrophils appears to be accomplished by inhibition of $Ca^{2+}$ influx from the extracellular medium.

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Effects of Dietary Supplementation of Fermented Chitin-chitosan (FERMKIT) on Toxicity of Mycotoxin in Ducks

  • Khajarern, J.M.;Khajarern, S.;Moon, T.H.;Lee, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.5
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    • pp.706-713
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    • 2003
  • Two experiments were conducted to evaluate the efficacy of dietary FERMKIT, a commercial toxin binder consisting of probiotic-fermented natural product containing chitin, chitosan and chitosan oligosaccharides ($FERMKITO^{(R)}$, EASY-BIO SYSTEM, Inc., Korea), in binding aflatoxin (AF) and zearalenone (ZEN) and ameliorating their mycotoxicity in meat type ducks. FERMKIT was supplemented to AF contaminated diets (at 120 ppb) at either 0.3 or 0.6% in experiment 1 and to ZEN contaminated diets (at 150 ppb) at 0.6% in experiment 2. In experiment 1 body weight gains were reduced by 37% and mortality was increased by 18% in ducks fed diet contaminated with AF at 120 ppb compared to ducks fed control diet (<10 ppb AF) for the 4-wk experimental period. However, dietary FERMKIT supplementation effectively alleviated overall toxicity induced by AF. The significant treatment-related changes in feather growth, web-toe hemorrhage, leg deformity, liver paleness, organ weights, hematological values and serum biochemical values, as compared to the control, were observed. The FERMKIT supplementation significantly diminished the adverse effects of AF and restored all the parameters measured back (<0.05) toward the control values. These findings indicated that FERMKIT, when added at the levels of 0.3 or 0.6% in the 120 ppb AF diets, could modulate the toxicity of AF with percentage sorption capacity of 52.70% at the level 0.3% and 79.85% at the level 0.6% of the diets (experiment 1). In experiment 2, FERMKIT, when added at 0.6% to the 150 ppb ZEN diets for the 4-wk experimental period, diminished the toxicity as shown by body weight gain, weights of testicles, oviducts, Bursa of Fabricius and cloaca eversion score as compared with the controls (<10 ppb ZEN) and 150 ppb ZEN diet with no added FERMKIT. The findings indicated that FERMKIT could be protective against the effects of ZEN in young growing ducks with percentage sorption capacity of 67.11% as evaluated from toxicity index parameter measured when added at 0.6% of the diets containing 150 ppb ZEN.

Effect of Oral Probiotics (Bifidobacterium lactis AD011 and Lactobacillus acidophilus AD031) Administration on Ovalbumin-Induced Food Allergy Mouse Model

  • Kim, Ji-Yeun;Choi, Young-Ok;Ji, Geun-Eog
    • Journal of Microbiology and Biotechnology
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    • v.18 no.8
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    • pp.1393-1400
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    • 2008
  • Recent study has demonstrated an increasing prevalence of food allergy in Korean children. Specific probiotic bacteria may promote potentially anti-allergenic processes through induction of Th1-type immunity and enhance the regulatory lymphocyte. This study investigated whether orally administrated probiotics could suppress allergic responses in an ovalbumin (OVA)-induced allergy mouse model. Thus, female C3H/HeJ mice were orally sensitized with OVA and cholera toxin for 4 weeks. Lactobacillus acidophilus AD031, Bifidobacterium lactis AD011, and L. acidophilus AD031 plus B. lactis AD011 were fed to mice from 2 weeks before the sensitization. The OVA-induced mice that were not treated with probiotics had significantly increased serum levels of OVA-specific IgE and IgG1, and OVA-specific IgA in feces. However, the mice treated with probiotics suppressed production of the OVA-specific IgE, IgG1, and IgA. The level of IL-4 was significantly lower, and the levels of INF-$\gamma$ and IL-10 were significantly higher in the mice treated with probiotics than that in the non-treated mice. The groups treated with probiotics had decreased levels of degranulated mast cells, eosinophil granules, and tail scabs. These results indicate that L. acidophilus AD031 and B. lactis AD011 might be useful for the prevention of allergy.

Lipoteichoic Acid from Lactobacillus plantarum Inhibits the Expression of Platelet-Activating Factor Receptor Induced by Staphylococcus aureus Lipoteichoic Acid or Escherichia coli Lipopolysaccharide in Human Monocyte-Like Cells

  • Kim, Hangeun;Jung, Bong Jun;Jeong, Jihye;Chun, Honam;Chung, Dae Kyun
    • Journal of Microbiology and Biotechnology
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    • v.24 no.8
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    • pp.1051-1058
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    • 2014
  • Platelet-activating factor receptor (PAFR) plays an important role in bacterial infection and inflammation. We examined the effect of the bacterial cell wall components lipopolysaccharide (LPS) and lipoteichoic acid (LTA) from Lactobacillus plantarum (pLTA) and Staphylococcus aureus (aLTA) on PAFR expression in THP-1, a monocyte-like cell line. LPS and aLTA, but not pLTA, significantly increased PAFR expression, whereas priming with pLTA inhibited LPS-mediated or aLTA-mediated PAFR expression. Expression of Toll-like receptor (TLR) 2 and 4, and CD14 increased with LPS and aLTA treatments, but was inhibited by pLTA pretreatment. Neutralizing antibodies against TLR2, TLR4, and CD14 showed that these receptors were important in LPS-mediated or aLTA-mediated PAFR expression. PAFR expression is mainly regulated by the nuclear factor kappa B signaling pathway. Blocking PAF binding to PAFR using a PAFR inhibitor indicated that LPS-mediated or aLTA-mediated PAF expression affected TNF-${\alpha}$ production. In the mouse small intestine, pLTA inhibited PAFR, TLR2, and TLR4 expression that was induced by heat-labile toxin. Our data suggested that pLTA has an anti-inflammatory effect by inhibiting the expression of PAFR that was induced by pathogenic ligands.

Isolation, Screening and Identification of Swine Gut Microbiota with Ochratoxin A Biodegradation Ability

  • Upadhaya, Santi Devi;Song, Jae-Yong;Park, Min-Ah;Seo, Ja-Kyeom;Yang, Liu;Lee, Chan-Ho;Cho, Kyung-J.;Ha, Jong-K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.1
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    • pp.114-121
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    • 2012
  • The potential for ochratoxin A (OTA) degradation by swine intestinal microbiota was assessed in the current study. Intestinal content that was collected aseptically from swine was spiked with 100 ppb OTA and incubated for 6 and 12 h at $39^{\circ}C$. An OTA assay was conducted using the incubated samples, and it was found that 20% of the OTA toxin was detoxified, indicating the presence of microbes capable of OTA degradation. Twenty-eight bacterial species were isolated anaerobically in M 98-5 media and 45 bacterial species were isolated using nutrient broth aerobically. Screening results showed that one anaerobic bacterial isolate, named MM11, detoxified more than 75% of OTA in liquid media. Furthermore, 1.0 ppm OTA was degraded completely after 24 h incubation on a solid 'corn' substrate. The bacterium was identified by 16S rDNA sequencing as having 97% sequence similarity with Eubacterium biforme. The isolation of an OTA-degrading bacterium from the swine natural flora is of great importance for OTA biodegradation and may be a valuable potential source for OTA-degradation enzymes in industrial applications.

The Phylogenetic Relationship of Several Oscillatorian Cyanobacteria, Forming Blooms at Daecheong Reservoirs, Based on Partial 16S rRNA Gene Sequences

  • Lee, Wook-Jae;Bae, Kyung-Sook
    • Journal of Microbiology and Biotechnology
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    • v.11 no.3
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    • pp.504-507
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    • 2001
  • The partial 16S gene sequences of six filamentous cyanobacterial strains, Oscillatoria lmosa KCTC AG10168, Oscillatoria princeps KCTC AG10153, Oscillatoria sp. KCTC AG 10184, Phormidium tenue KCTC AG10158, Phormidium parchydematicum KCTC AG10164, and Lyngbya hieronymusii KCTC AG10199, which were isolated in the late summer at Daecheong Reservoirs, were determined and assigned their phylogenetic and taxonomic position among taxa of order Ocillatoriales whose partial 16S rRNA gene sequences aligned in this suty, were very heterogeneously clustered with other taxa. The two strains, Oscillatoria limosa KCTC AG10168 and O. princeps KCTC AG10153, formed a cluster with O. sancta PCC7515, which supported 64% of the bootstrap trees with high similarity (19-96.15%). Strain Oscillatoria sp. KCTC AG10184, that was known to produce a nasty substance, was closely related to the toxic Oscillatoria group. The study on morphological variation in various environments and toxin production will confirm the taxonomic status of these species. Phormidium tenus KCTC AG10158 and Phormidium parchydematicum KCTC AG10164 made a cluster with other oscillatorian species of Phormidium, Oscillatoria, and Leptolynbya, which supproted 100% of the bootstrap trees with a very high sequence smilarity (96.8-99.8%) in thsi study. The sequence analysis in this study also supported that taxa of oscillatoriales are not monophyletic. Some of the fractures, such as the presence or absence of sheath and cell shape, which were used to define them, would be inadequate and should be reconfirmed. We suggest that sequences of partial 16S rRNA gene fragments aligned in this study should be more useful than morphological features in the identification and reconfirmation of the taxonomic status of these oscillactorian cyanobacteria.

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Investigation of Quorum Sensing-Dependent Gene Expression in Burkholderia gladioli BSR3 through RNA-seq Analyses

  • Kim, Sunyoung;Park, Jungwook;Choi, Okhee;Kim, Jinwoo;Seo, Young-Su
    • Journal of Microbiology and Biotechnology
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    • v.24 no.12
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    • pp.1609-1621
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    • 2014
  • The plant pathogen Burkholderia gladioli, which has a broad host range that includes rice and onion, causes bacterial panicle blight and sheath rot. Based on the complete genome sequence of B. gladioli BSR3 isolated from infected rice sheaths, the genome of B. gladioli BSR3 contains the luxI/luxR family of genes. Members of this family encode N-acyl-homoserine lactone (AHL) quorum sensing (QS) signal synthase and the LuxR-family AHL signal receptor, which are similar to B. glumae BGR1. In B. glumae, QS has been shown to play pivotal roles in many bacterial behaviors. In this study, we compared the QS-dependent gene expression between B. gladioli BSR3 and a QS-defective B. gladioli BSR3 mutant in two different culture states (10 and 24 h after incubation, corresponding to an exponential phase and a stationary phase) using RNA sequencing (RNA-seq). RNA-seq analyses including gene ontology and pathway enrichment revealed that the B. gladioli BSR3 QS system regulates genes related to motility, toxin production, and oxalogenesis, which were previously reported in B. glumae. Moreover, the uncharacterized polyketide biosynthesis is activated by QS, which was not detected in B. glumae. Thus, we observed not only common QS-dependent genes between B. glumae BGR1 and B. gladioli BSR3, but also unique QS-dependent genes in B. gladioli BSR3.

Evaluation of the Probiotic Potential of Bacillus polyfermenticus CJ6 Isolated from Meju, a Korean Soybean Fermentation Starter

  • Jung, Ji Hye;Lee, Myung Yul;Chang, Hae Choon
    • Journal of Microbiology and Biotechnology
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    • v.22 no.11
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    • pp.1510-1517
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    • 2012
  • To evaluate the probiotic potential of Bacillus polyfermenticus CJ6 isolated from meju, a Korean traditional soybean fermentation starter, its functionality and safety were investigated. B. polyfermenticus CJ6 was sensitive to all antibiotics listed by the European Food Safety Authority. The strain was also non-hemolytic, carried no emetic toxin or enterotoxin genes, and produced no enterotoxins. The resistance of B. polyfermenticus CJ6 vegetative cells and spores to simulated gastrointestinal conditions was high (60-100% survival rate). B. polyfermenticus CJ6 produced high amounts (0.36 g as a purified lyophilized form) of ${\gamma}$-polyglutamic acid (PGA). We speculate that the improved cell viability and the production of ${\gamma}$-PGA have a significant correlation. Adhesion of the strain to Caco-2 and HT-29 cells was weaker than that of the reference strain (Lb. rhamnosus GG), but it was comparable to or stronger than those of reported Bacillus spp. When B. polyfermenticus CJ6 spores were given orally to mice, the number of cells excreted in the feces was 4-fold higher than the original inocula. This suggests the inoculated spores propagated within the intestinal tract of the mice. This idea was confirmed by field emission scanning electron microscopy, which revealed directly that B. polyfermenticus CJ6 cells germinated and adhered within the gastrointestinal tract of mice. Taken together, these findings suggest that B. polyfermenticus CJ6 has probiotic potential for both human consumption and use in animal feeds.