• Title/Summary/Keyword: H1-2 toxin

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Shiga toxin-associated hemolytic uremic syndrome complicated by intestinal perforation in a child with typical hemolytic uremic syndrome

  • Chang, Hye Jin;Kim, Hwa Young;Choi, Jae Hong;Choi, Hyun Jin;Ko, Jae Sung;Ha, Il Soo;Cheong, Hae Il;Choi, Yong;Kang, Hee Gyung
    • Clinical and Experimental Pediatrics
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    • v.57 no.2
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    • pp.96-99
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    • 2014
  • Hemolytic uremic syndrome (HUS) is one of the most common causes of acute renal failure in childhood and is primarily diagnosed in up to 4.5% of children who undergo chronic renal replacement therapy. Escherichia coli serotype O157:H7 is the predominant bacterial strain identified in patients with HUS; more than 100 types of Shiga toxin-producing enterohemorrhagic E. coli (EHEC) subtypes have also been isolated. The typical HUS manifestations are microangiopathic hemolytic anemia, thrombocytopenia, and renal insufficiency. In typical HUS cases, more serious EHEC manifestations include severe hemorrhagic colitis, bowel necrosis and perforation, rectal prolapse, peritonitis, and intussusceptions. Colonic perforation, which has an incidence of 1%-2%, can be a fatal complication. In this study, we report a typical Shiga toxin-associated HUS case complicated by small intestinal perforation with refractory peritonitis that was possibly because of ischemic enteritis. Although the degree of renal damage is the main concern in HUS, extrarenal complications should also be considered in severe cases, as presented in our case.

Antimicrobials Effective for Inhibition of Enterohemorrhagic Escherichia coli Strains O26, O111, and O157 and Their Effects on Shiga Toxin Releases

  • Lee, John-Hwa;Stein, Barry D.
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1238-1243
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    • 2009
  • The susceptibilities of major enterohemorrhagic Escherichia coli (EHEC) strains to antimicrobial agents and the cytotoxicity of these agents were examined using a total of 38 strains of E. coli O26, O111, and O157, which are the major serogroups of EHEC. Among the 38 strains, 35, 36, and 36 were susceptible to amikacin, imipenem, and norfloxacin, respectively. These antimicrobial agents were further examined to determine their cytotoxicity on Vero cells as well as their effect on the release of Shiga toxins along with trimethoprim/sulfamethoxazole. Each of the E. coli O26, O111, and O157 strains containing both the stx1 and stx2 genes were grown in the absence or presence of these agents at 1/4 minimal inhibitory concentration for 6 h, 12 h, and 18 h. At the concentrations used in this study, none of the agents significantly altered cell count compared with the control group. The level of cytotoxicity in the imipenem group was lower at 12 hand 18 h than their respective controls. In contrast, the level of cytotoxicity in cultures treated with trimethoprim/sulfamethoxazole, norfloxacin, and amikacin was increased. The strains were also examined for the release of Shiga toxins 1 and 2 following treatment with the agents, which were measured by the reversed passive latex agglutination (RPLA) method. The RPLA assay showed a suppression of release of Shiga toxin 2 in the strain cultures containing imipenem. These results indicate that imipenem may be a safe and effective agent for inhibition of these bacteria, which has clinical implications for the treatment of EHEC infections.

Killer Characteristics of Candida dattila KI09 and Kl12 Strains (Candida dattila K109와 K112 균주의 Killer 특성)

  • 정원철;장해춘;최언호
    • Microbiology and Biotechnology Letters
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    • v.18 no.1
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    • pp.26-30
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    • 1990
  • Candida dattila K109 and K112 isolated from grapes in Korea showed killer activity toward Saccharomyces cereuisiae 5 x 47, S. cereuisiae 1368, Hamenula, Torulopsis, Kluyueromyces, Debaryomyces, and Brettanomyces, and showed the most effective killer activity at 22-26$^{\circ}C$ and at pH 3.9-4.1. The killer actvity of both toxins were remarkably decreased at higher temperature than $25^{\circ}C$ and higher pH than pH 4.0. And the toxins were suggested to be glycoproteins inactivated by pronase E and pepsin. The killer activity was not cured by incubation at elevated temperature of 30-37"C, but cured by treatment with 0.0105-0.3 ppm cycloheximie.imie.

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Molecular Dissection of the Interaction between hBLT2 and the G Protein Alpha Subunits

  • Vukoti, Krishna Moorthy;Lee, Won-Kyu;Kim, Ho-Jun;Kim, Ick-Young;Yang, Eun-Gyeong;Lee, Cheol-Ju;Yu, Yeon-Gyu
    • Bulletin of the Korean Chemical Society
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    • v.28 no.6
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    • pp.1005-1009
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    • 2007
  • Leukotriene B4 (LTB4) is a potent chemoattractant for leukocytes and considered to be an inflammatory mediator. Human BLT2 (hBLT2) is a low-affinity G-protein coupled receptor for LTB4 and mediates pertussis toxin-sensitive chemotactic cell movement. Here, we dissected the interaction between hBLT2 and G-protein alpha subunits using GST fusion proteins containing intracellular regions of hBLT2 and various Gα protein including Gα i1, Gα i2, Gα i3, Gα s1, Gα o1, and Gα z. Among the tested Gα subunits, Gα i3 showed the highest binding to the third intracellular loop region of hBLT2 with a dissociation constant (KD) of 5.0 × 10?6 M. These results suggest that Gα i3 has the highest affinity to hBLT2, and the third intracellular loop region of hBLT2 is the major component for the interaction with Gα i3.

3-Methylthiopropionic Acid of Rhizoctonia solani AG-3 and Its Role in the Pathogenicity of the Fungus

  • Kankam, Frederick;Long, Hai-Tao;He, Jing;Zhang, Chun-hong;Zhang, Hui-Xiu;Pu, Lumei;Qiu, Huizhen
    • The Plant Pathology Journal
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    • v.32 no.2
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    • pp.85-94
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    • 2016
  • Studies were conducted to determine the role of 3-methylthioproprionic acid (MTPA) in the pathogenicity of potato stem canker, Rhizoctonia solani, and the concentrations required to inhibit growth of R. solani under laboratory and plant house-based conditions. The experiments were laid out in a completely randomized design with five treatments and five replications. The treatments were 0, 1, 2, 4, and 8 mM concentrations of MTPA. The purified toxin exhibited maximal activity at pH 2.5 and $30^{\circ}C$. MTPA at 1, 2, 4, and 8 mM levels reduced plant height, chlorophyll content, haulm fresh weight, number of stolons, canopy development, and tuber weight of potato plants, as compared to the control. MTPA significantly affected mycelial growth with 8 mM causing the highest infection. The potato seedlings treated with MTPA concentrations of 1.0-8.0 mM induced necrosis of up to 80% of root system area. Cankers were resulted from the injection of potato seedling stems with 8.0 mM MTPA. The results showed the disappearance of cell membrane, rough mitochondrial and cell walls, change of the shape of chloroplasts, and swollen endoplasmic reticulum. Seventy-six (76) hours after toxin treatment, cell contents were completely broken, cytoplasm dissolved, and more chromatin were seen in the nucleus. The results suggested that high levels of the toxin concentration caused cell membrane and cytoplasm fracture. The integrity of cellular structure was destroyed by the phytotoxin. The concentrations of the phytotoxin were significantly correlated with pathogenicity and caused damage to the cell membrane of potato stem base tissue.

Effect of pertussis toxin pretreated centrally on blood glucose level induced by stress

  • Suh, Hong-Won;Sim, Yun-Beom;Park, Soo-Hyun;Sharma, Naveen;Im, Hyun-Ju;Hong, Jae-Seung
    • The Korean Journal of Physiology and Pharmacology
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    • v.20 no.5
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    • pp.467-476
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    • 2016
  • In the present study, we examined the effect of pertussis toxin (PTX) administered centrally in a variety of stress-induced blood glucose level. Mice were exposed to stress after the pretreatment of PTX (0.05 or 0.1 mg) i.c.v. or i.t. once for 6 days. Blood glucose level was measured at 0, 30, 60 and 120 min after stress stimulation. The blood glucose level was increased in all stress groups. The blood glucose level reached at maximum level after 30 min of stress stimulation and returned to a normal level after 2 h of stress stimulation in restraint stress, physical, and emotional stress groups. The blood glucose level induced by cold-water swimming stress was gradually increased up to 1 h and returned to the normal level. The intracerebroventricular (i.c.v.) or intrathecal (i.t.) pretreatment with PTX, a $G_i$ inhibitor, alone produced a hypoglycemia and almost abolished the elevation of the blood level induced by stress stimulation. The central pretreatment with PTX caused a reduction of plasma insulin level, whereas plasma corticosterone level was further up-regulated in all stress models. Our results suggest that the hyperglycemia produced by physical stress, emotional stress, restraint stress, and the cold-water swimming stress appear to be mediated by activation of centrally located PTX-sensitive G proteins. The reduction of blood glucose level by PTX appears to due to the reduction of plasma insulin level. The reduction of blood glucose level by PTX was accompanied by the reduction of plasma insulin level. Plasma corticosterone level up-regulation by PTX in stress models may be due to a blood glucose homeostatic mechanism.

Optimal Cultur Conditions for the Production of Insecticidal Toxin by Xenorhabdus nematophilus Isolated from Steinernema carpocapsae (Steinernema carpocapsae로부터 분리된 Xenorhabdus nematophilus에 의한 살충물질 생산을 위한 최적 배양조건)

  • 유연수;박선호
    • KSBB Journal
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    • v.15 no.1
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    • pp.100-105
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    • 2000
  • Optimal medium composition, culture conditions, characteristics of phase variation and activity of insecticidal toxin by Xenorhabdus nematophilus isolated and identified from Korean entomopathogenic nematode Steinernema carpocapsae were examined. Optimal medium composition of this strain was 50-70 g/L yeast extract, 3 g/L $K_{2}HPO_{4}$, 1g/L $NH_{4}H_{2}PO_{4}$, 2g/L ${MgSO}_4$$\cdot$${7H}_{2}O$, 10g/L NaCl and, these, yeast extract was found as a limiting nutrient for cell growth. When Monod equation was applied, maxmum specific growth rate and Monod constant were estimated as 0.13 $hr^{-1}$ and 20g/L, respectively. The pH of culture medium increased up to 8.5-9.5 regardless of initial pH 6-7 as the cells continued to grow. The specific growth rate in a 7 L fermentor was 0.18 $hr^{-1}$, which was enhancement 1.4 fold compared to a flask culture. In case of phase variation, phase I fraction was maintained above 90% at the stationary phase for both flask and fermentor cultures. According to oral toxicity test of Gallena mellonella by Xenorhabdus nematophilus, the addition of cell pellets into feed inhibited normal growth of insect larvae and killed completely then after 20 days cultivation. When culture supernatant of this strain was injected into hemolymph of insect larva, the toxicity was strongest at 24hr cultivation in the early exponential phase and gradually decreased as the culture time proceeded.

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Relationship between Gb3 Expression and Cytotoxicity of Shiga-like Toxin I (Shiga-like Toxin I의 세포독성과 수용체 Gb3 발현과의 관계)

  • Lim, Suk-Hwan;Kim, Gi-Young;Kim, Hyung-Chun;Kim, Young-Hee;Son, Yong-Hae;Oh, Yang-Hyo;Park, Yeong-Min
    • Clinical and Experimental Pediatrics
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    • v.46 no.2
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    • pp.143-153
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    • 2003
  • Purpose : Infection with Shiga-like toxin (SLT)-producing Escherichia coli, an emerging human pathogen found particularly in young children under 5 years of age, causes a spectrum of illnesses with high morbidity and mortality, ranging from diarrhea to hemorrhagic colitis and hemolytic uremic syndrome. Host mediators play an important role in the pathogenesis of SLT-I toxicity. The experiments described here were designed to investigate the effect of SLT-I on TNF-${\alpha}$ production and to understand the effect of TNF-${\alpha}$ on GB3 expression. We also further examine the relationship between the Gb3 level and the differential susceptibility of cells to the cytotoxic action of SLT-I. Methods : The effect of purified SLT-1 from E. coli O157 : H7 (ATCC 43890) on tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) production in Raw264.7 cells was investigated. Many mediators regulate endothelial cell membrane expression of the glycolipid globotriaosyleramide (Gb3), which serves as the toxin receptor, suggesting that the host response to the toxin or other bacterial products may contribute to pathogenesis by regulating target cell sensitivity to the toxins. Therefore, the relationships between Gb3 expression and cytotoxicity against SLT-I on three types of cells were evaluated. Results : Detectable levels of TNF-${\alpha}$ were produced as early as six hours after induction and continued to increase during 48 hours by SLT-I. It was also found that Vero cells and dendritic cells (DC2.4 cells) expressed high levels of Gb3, 83% and 68%, respectively, and that Raw264.7 cells had a low level of Gb3 (29%) and appeared refractory to cytotoxicity against SLT-I. Vero cells and DC2.4 cells expressing high levels of Gb3 were highly susceptible to SLT-I. Furthermore, macrophages showed a resistance to SLT-I cytotoxicity, despite the fact that Gb3 expression was enhanced. Conclusion : These results strongly suggest that the expression of Gb3 is necessary but not sufficient to confer sensitivity of macrophages to SLT-I and further underpin the important role of SLT-I and its Gb3 receptors in the pathogenesis of E. coli O157 infection.

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.

The Cytotoxic Mechanisms of Bacillus thuringiensis $\delta$-endotoxin, a Bioinsecticide : Effect on $K^+$ Channel of Insect Cell Lines.

  • Seo, Young-Rok;Han, Sung-Sik;Yu, Yong-Man;Lee, Jun-Jae;Ryu, Jae-Chun
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 1996.12a
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    • pp.70-70
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    • 1996
  • The cytotoxicological effect of Bt 1-endotoxin, well-known as a bioinsecticide, was investigated on ion channel of insect cell lines. This study attempted to evaluted the specificity by simple experiment to measure the cell swelling using lepidopteran cell lines in isotonic solution containing only one cation. Cell swelling was stimulated in KCI-sucrose isotonic solution as well as TC-100 media containg in solubilized crystal 5-endotoxin. It suggested that the cell swelling by Bt toxin have a relation to K+ channel. The cell swelling may be due to the stimulation K+ influx and simultaneously the penetration of H2O induced by Bt toxin, because the stimulation of swelling was observed with the solubilized toxin in KCI-sucrose isotonic solution, but not in sucrose isotonic solution. Moreover the specific K+ channel blocker, such as 4-arnjnopyrimidine(4-AP) and ouabain, showed the significant effect on the cell swelling induced by Bt toxin. The increasement of the cell swelling induced by 4-AP suggested to be caused by the block of K+ efflux through K+ leak channels. The inhibition of cell swelling by ouabain, which is the well-known inhibitor of Na+, K+-ATPase, suggested to be due to decreasement of K+ influx following diminishment of Na+, K+-ATPase activities.

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