• Title/Summary/Keyword: T-2 Toxin

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Helicobacter pylori Vacuolating Toxin Exhibits Polar Activity of $Cl^-$ Secretion and Secretory Response to Carbachol in T84 Cells

  • Jin, Nan-Ge;Jin, Yong-Ri;So, In-Suk;Kim, Ki-Whan
    • The Korean Journal of Physiology and Pharmacology
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    • v.8 no.5
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    • pp.289-293
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    • 2004
  • To investigate whether VacA (vacuolating toxin) produced by Helicobacter pylori Korean stain 99 induces intestinal secretion, purified VacA was added to T84 cell monolayers mounted in Ussing chambers, and electrical parameters were monitored. Mucosal addition of low pH-pretreated VacA increased short circuit current (Isc). The effect was time- and dose-dependent and saturable. The time-to-peak Isc was concentration-dependent. Chloride channel inhibitors, niflumic acid or 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB), inhibited VacA-stimulated Isc. Carbachol (CCh)-induced increase of Isc was prolonged by the addition of VacA to the mucosal side only. The effect was unaltered by the addition of niflumic acid. VacA did not show cytopathic effects. These studies indicate that VacA is a nonlethal toxin that acts in a polar manner on T84 monolayers to potentiate $Cl^-$ secretion and the response to CCh secretion without decrease in monolayer resistance. VacA may contribute to diarrhea diseases in human intestinal epithelial cells.

Analysis of the Apoptotic Mechanisms of Snake Venom Toxin on Inflammation-induced HaCaT Cell-line

  • Chun, Youl Woong;Song, Ho Sueb
    • Journal of Acupuncture Research
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    • v.34 no.1
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    • pp.23-30
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    • 2017
  • Objectives : In this study, the roles of Interleukin (IL)-4 and Signal transducer and activator of transcription 6 (STAT6), which have been reported to play a role in the pathogenesis of inflammation and cancer, were evaluated in snake venom toxin (SVT)-induced apoptosis. Methods : Inflammation was induced in human HaCaT kerationocytes, by lipopolysaccharide (LPS; $1{\mu}g/mL$) or tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), followed by treatment with SVT (0, 1, or $2{\mu}g/mL$). Cell viability was assessed by MTT assays after 24 h, and the expression of levels of IL-4, STAT6, and the apoptosis-related proteins p53, Bax, and Bcl-2 were evaluated by western blotting. Electro mobility shift assays (EMSAs) were performed to evaluate the DNA binding capacity of STAT6. Results : MTT assays showed that inflammation-induced growth of HaCaT cells following LPS or TNF-${\alpha}$ stimulation was inhibited by SVT. Western blot analysis showed that p53 and Bax, which promote apoptosis, were increased, whereas that of Bcl-2, an anti-apoptotic protein, was decreased in a concentration-dependent manner in LPS- or TNF-${\alpha}$-induced HaCaT cells following treatment with SVT. Moreover, following treatment of HaCaT cells with LPS, IL-4 concentrations were increased, and treatment with SVT further increased IL-4 expression in a concentration-dependent manner. Western blotting and EMSAs showed that the phosphorylated form of STAT6 was increased in HaCaT cells in the context of LPS- or TNF-${\alpha}$-induced inflammation in a concentration-dependent manner, concomitant with an increase in the DNA binding activity of STAT6. Conclusion : SVT can effectively promote apoptosis in HaCaT cells in the presence of inflammation through a pathway involving IL-4 and STAT6.

Occurrence of Mycotoxins in Korean Grains and Their Simultaneous Analysis (한국산 곡류에서의 곰팡이독소 오염현황 및 동시분석)

  • Kim, Dong-Ho;Jang, Han-Sub;Choi, Gyu-Il;Kim, Hyun-Jung;Kim, Ho-Jin;Kim, Hyo-Lin;Cho, Hyun-Jung;Lee, Chan
    • Korean Journal of Food Science and Technology
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    • v.45 no.1
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    • pp.111-119
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    • 2013
  • Eleven mycotoxins, including aflatoxins, ochratoxin A, fumonisins, zearalenone, T-2 toxin, deoxynivalenol, and HT-2 toxin, were analyzed simultaneously in rice, barley, and maize produced in 2011 by liquid chromatography coupled with triple quadrupole mass spectrometry (LC/MS/MS). Limits of detection (LOD) are 0.2 ${\mu}g/kg$ for aflatoxin $B_1$, and $G_1$, 0.3 ${\mu}g/kg$ for aflatoxins $B_2$, and $G_2$, 0.1 ${\mu}g/kg$ for ochratoxin, fumonisins, zearalenone, and T-2 toxin and 3.0 ${\mu}g/kg$ for deoxynivalenol and HT-2 toxin. Limits of quantification (LOQ) were 0.6 ${\mu}g/kg$ for aflatoxins $B_1$, and $G_1$, 0.9 ${\mu}g/kg$ for aflatoxins $B_2$, and $G_2$, 0.3 ${\mu}g/kg$ for ochratoxin, fumonisins, zearalenone, and T-2 toxin and 10.0 ${\mu}g/kg$ for deoxynivalenol and HT-2 toxin. Recoveries for 11 mycotoxins ranged from 70.45 to 111.11%. Fumonisins, deoxynivalenol, and zaeralenone were detected from 0.9 to 334.0 ${\mu}g/kg$ in the polished rice, barley and raw corn cultivated in Korea. Other mycotoxins were not detected. Deoxynivalenol contamination was mainly found in barley (24 out of 43 samples) and the average value in positive samples was 113.30 ${\mu}g/kg$.

Analysis of Transgenic Mouse, for the Production of Immunodeficiency Animals (면역결핍동물의 생산을 위한 형질전환생쥐의 분석)

  • 나루세겐지;양정희;이승현;최화식;이성호;박창식;진동일
    • Korean Journal of Animal Reproduction
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    • v.27 no.2
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    • pp.179-185
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    • 2003
  • To determine whether the diphtheria toxin-A (DT) gene disrupts development of thymocytes in transgenic animal, the DT-A gene was used for the production of transgenic mice directed by proximal Ick promoter sequences. Two transgenic founder mice that contained several copies of transgene were produced by DNA microinjection and integration of transgene in transgenic mice was confirmed by PCR and Southern blotting analysis. Transgenic $F_1$ and $F_2$ mice were produced by outbreeding of founder and $F_1$ mice to investigate expression of transgene and phenotypes in transgneic mice. Expression of the diphtheria toxin gene was confirmed in thymus, spleen and liver of transgenic mice by RT-PCR. In circulating blood of transgenic mice, lower number of circulating white blood cells and platelets were observed compared with that of normal mice. In addition, transgneic mice had reduced number of circulating peripheral T-cells analyzed by FACS with anti-CD3 antibody. The data in these transgenic mice indicate that DT gene can play a disruptive role in developing thymocytes of transgenic mice resulted in lower number of T-cells that can be applicable to a wide range of tissues in other animals.

Synthetic Cannabinoid-Induced Immunosuppression Augments Cerebellar Dysfunction in Tetanus-Toxin Treated Mice

  • Yun, Jaesuk;Gu, Sun Mi;Lee, Tac-hyung;Song, Yun Jeong;Seong, Seonhwa;Kim, Young-Hoon;Cha, Hye Jin;Han, Kyoung Moon;Shin, Jisoon;Oh, Hokyung;Jung, Kikyung;Ahn, Chiyoung;Park, Hye-Kyung;Kim, Hyung Soo
    • Biomolecules & Therapeutics
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    • v.25 no.3
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    • pp.266-271
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    • 2017
  • Synthetic cannabinoids are one of most abused new psychoactive substances. The recreational use of abused drug has aroused serious concerns about the consequences of these drugs on infection. However, the effects of synthetic cannabinoid on resistance to tetanus toxin are not fully understood yet. In the present study, we aimed to determine if the administration of synthetic cannabinoids increase the susceptibility to tetanus toxin-induced motor behavioral deficit and functional changes in cerebellar neurons in mice. Furthermore, we measured T lymphocytes marker levels, such as CD8 and CD4 which against tetanus toxin. JWH-210 administration decreased expression levels of T cell activators including cluster of differentiation (CD) $3{\varepsilon}$, $CD3{\gamma}$, CD74p31, and CD74p41. In addition, we demonstrated that JWH-210 induced motor impairment and decrement of vesicle-associated membrane proteins 2 levels in the cerebellum of mice treated with tetanus toxin. Furthermore, cerebellar glutamatergic neuronal homeostasis was hampered by JWH-210 administration, as evidenced by increased glutamate concentration levels in the cerebellum. These results suggest that JWH-210 may increase the vulnerability to tetanus toxin via the regulation of immune function.

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.

Toxicity of the Puffer fish, Takifugu pardalis (Jolbok) and Takifugu niphobles (Bokseom) from Coastal Area of Korea (한국 연안산 졸복(Takifugu pardalis)과 복섬(Takifugu niphobles)의 독성)

  • Kim, Ji-Hoe;Son, Kwang-Tae;Mok, Jong-Soo;Oh, Eun-Gyoung;Hwang, Hye-Jin;Yu, Hong-Sik;Lee, Hee-Jung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.40 no.5
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    • pp.269-275
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    • 2007
  • The toxicity of two species of puffer fish, Takifugu pardalis and T. niphobles, collected from the coastal regions of Korea was determined using a mouse bioassay. In T. pardalis collected at Tongyeong, the proportion of toxic specimens containing ${\geq}10MU/g$ exceeded 90% for the skin, fins, liver, intestine, ovary, and gallbladder, 11.1% for the testis, and 6.9% for the muscle. In each of the organs, the highest toxin levels were several tens (14-39) of mouse units (MU) per gram in the muscle, testis, and eyeball, but thousands (1,444-5,755) of MU per gram in the skin, liver, intestine, ovary, and gallbladder. The organs of T. pardalis exhibited remarkable variation in toxicity. In T. niphobles, the proportion of toxic specimens exceeded 90% for the ovary and skin, 60-80% for the fins, liver, intestine, and gallbladder, and 4.5% for the muscle; no toxicity was detected in the testis or eyeball using the mouse bioassay. The highest toxin levels were thousands (2,291-7,777) of MU per gram in the liver, intestine, ovary, and gallbladder, hundreds(146-328) of MU per gram in the skin and fins, and 18 MU/g in the muscle. Takifugu niphobles toxicity also exhibited remarkable regional variation. The toxicity in the edible muscle of T. pardalis and T. niphobles was at acceptable levels for human consumption, while the toxicity of the skin of both species of puffer fish was very high, so that care must be taken when used for human consumption.

Effects of Minor Arginyl tRNA and Isoleucyl tRNA on the Expression of Clostridium botulinum Neurotoxin Light Chain in Escherichia coli

  • Kim, Jin-Sook;Seong, Hye-Young;Kim, Mi-Wha;Ku, Jong-Seo;Choi, Soon-Yong
    • Journal of Microbiology and Biotechnology
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    • v.13 no.2
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    • pp.287-291
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    • 2003
  • Botulinum neurotoxin type A (BONT/A) is an extremely potent toxin, which is produced by Clostridium botulinum. The light chain of this protein (BONT/A LC), which is known as a zinc endopeptidase, cleaves SNAP-25 involved in the exocytosis process. In this work, the expression of recombinant BoNT/A LC in E. coli is described. The BONT/A LC gene of C. botulinum contains a high frequency of the arginine AGA and isoleucine ATA codons that are rarely used in genes of E. coli, hampering the translation of recombinant protein. The argD and ilex tRNA genes were cloned into pACYC184 vector, resulting in pAAD131X plasmid. The translational stress of the toxin gene related to codon bias was reversed by fupplernentation of the AGA arginyl tRNA of T4 phage and AUA isoleucyl tRNA of E. coli. This system may be applicable for the expression of a variety of AT-rich heterologous genes in E. coli.

Diversity of Mycotoxigenic Fusarium armeniacum Isolated from Rice Grains at Harvest Time in Korea (수확기 벼 이삭에서 분리된 진균독소 생성 Fusarium armeniacum의 다양성)

  • Hong, Sung Kee;Lee, Soohyung;Lee, Theresa;Ham, Hyeonheui;Mun, Hye Yeon;Choi, Hyo Won;Son, Seung-Wan;Ryu, Jae-Gee
    • The Korean Journal of Mycology
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    • v.43 no.3
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    • pp.158-164
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    • 2015
  • A total of 509 rice panicle samples were collected at harvest time from fields in 8 provinces from 2010 to 2014. One hundred five grains per sample were plated on potato dextrose agar and 6,658 Fusarium isolates were obtained; among them, 67 were identified as Fusarium armeniacum by sequencing the translation elongation factor $1{\alpha}$ ($EF-1{\alpha}$) and confirmed by their morphological and cultural characteristics. Considerable variation in conidial size, colony color and $EF-1{\alpha}$ sequences was observed among the fungal isolates. The ability of 24 F. armeniacum isolates to produce T-2 and HT-2 toxin in potato sucrose agar was determined using liquid chromatography-mass spectrometry. Twenty one isolates produced T-2 and HT-2 toxin, resulting in varying toxin levels among the isolates. The results show that Korean isolates of F. armeniacum have diversity with respect to morphological, cultural, genetic, and toxigenic properties.

Simultaneous Analysis and Survey for Contamination of Nivalenol, Deoxynivalenol, T-2 toxin and Zearalenone in Feed (사료 중 Nivalenol, Deoxynivalenol, T-2 foxin과 Zearalenone의 동시분석과 오염도조사)

  • Kim, Dong-Ho;Kim, Hyun-Jung;Jang, Han-Sub;Kim, Yeong-Min;Choi, Heng-Bo;Ahn, Jong-Sung
    • Journal of Food Hygiene and Safety
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    • v.26 no.1
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    • pp.1-11
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    • 2011
  • Nivalenol (NIV), deoxynivalenol (DON), T-2 toxin (T-2) and zearalenone (ZEN) are mycotoxins produced by some Fusarium species known to be very frequently contaminated in feed. The study for simultaneous analysis and contamination survey in animal feed carried out. All mycotoxins were analysed by using high performance liquid chromatography tandem mass with internal standard. The limits of detection (LOD) were $2.0\;{\mu}g/kg$, $1.0\;{\mu}g/kg$, $1.0\;{\mu}g/kg$ and $0.1\;{\mu}g/kg$ for NIV, DON, T-2 and ZEN, respectively. Two hundred and thirty nine samples of feed were collected. The average concentration of DON was $212.3\;{\mu}g/kg$, $207.8\;{\mu}g/kg$ and $812.1\;{\mu}g/kg$ in chicken, pig and cattle feed, respectively. The average concentration of ZEN was $31.2\;{\mu}g/kg$, $35.6\;{\mu}g/kg$ and $147.2\;{\mu}g/kg$ for them, respectively. Especially, the levels of contamination for DON and ZEN were higher than those of NIV or T-2. And, the levels of contamination for four Fusarium mycotoxins in cattle feed appeared higher than those of pig and chicken feed. It was investigated that the high level of mycotoxin contamination in cattle feed was caused by com gluten feed of ingredients for feed, mainly.