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Omics for mechanisms of Edwardsiella tarda in resistance to plasma
S216
Abstracts / Toxicology Letters 205S (2011) S180–S300
haemolyticus as an antigen, VP1061 and VP2850 showed broadly cross-protective ability against V. alginolyticus, A. hydrophila and P. fluorescens in fish and mouse models. In V. alginolyticus as an antigen, the carps immunized with VA0764 and VA1061 had abilities to fight against infections not only caused by V. alginolyticus but also by A. hydrophila and P. fluorescens. These vaccine candidates are importance in against virulent bacterial infections. doi:10.1016/j.toxlet.2011.05.740
P2117 Responses of pulmonary proteins to paraquat in rats Q.X. Li 1,∗ , I.K. Cho 1 , M. Jeong 2 , J.K. Kim 2 , S.S. Hong 2 , K.H. Park 2 1
Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, Honolulu, USA, 2 Crop Life Safety, National Academy of Agricultural Science, Suwon-si, Republic of Korea
sub-proteome level to understand tolerance mechanism of bacteria to disinfectant. Methods: Two dimensional electrophoresis (2-DE) and mass spectrometry (MS) technologies were used to identify differential expression of OMPs of E. coli K-12 BW25113 in response to isopropanol stress. Gene knockout, complementation and overexpression were utilized to investigate the role of altered OM proteins. Results of the study: Up-regulation of LamB, FadL and OmpC, and down-regulation of OmpT, Tsx, OmpA, OmpF were detected. Furthermore, we investigated roles of these altered proteins in resistance to the disinfectant through gene knockout, complementation and overexpression. Our results showed that LamB, OmpA and OmpC played more important role in the resistance than the others. Finally, we characterized the ability of EnvZ/OmpR twocomponent signal transduction system in exposure to disinfectant. These results indicated the importance of OMPs and EnvZ/OmpR two-component signal transduction system in response to isopropanol. doi:10.1016/j.toxlet.2011.05.742
Purpose: Paraquat is among the most widely used herbicides in the world. Paraquat, when ingested, is very toxic to mammals including humans and may cause acute respiratory distress syndrome. The purpose of this study was to investigate the protein profiles in response to paraquat exposure. Methods: Sprague–Dawley male rats were orally administered with paraquat at 5, 10, 25 and 40 mg/kg bw (LD50 = 126 mg/kg) for five times a week for four weeks. Soluble proteins in pulmonary tissues were extracted and analyzed with a shotgun proteomic approach which combined one-dimensional gel electrophoresis, ion trap mass spectrometry and amino acid sequence alignment. Results: A number of 304 and 147 proteins were over- and under-represented, respectively, in the rat lung tissues after exposure to 25 mg/kg or higher of paraquat. The over-represented proteins include C-jun-amino-terminal kinase interacting protein 2, CDKN2A-intercting protein, mitogen-activated protein kinases, macrophage receptor with collagenous structure, complement decay-accelerating factor, stress-70 protein and thioredoxin reductase 3. All the over-and under-represented proteins were subjected to Ingenuity Pathway Analysis to create 25 biological networks and 38 pathways of interacting protein clusters according to the identifiers’ HomoloGene to the ortholog information in the Ingenuity Knowledge Base (Human/Mouse/Rat data). The responded pathways related to pulmonary stress include cardiovascular-cancerrespiratory pathway (e.g., nitric oxide synthase 3), regulation of clathrin-mediated endocytosis (e.g., AP2-associated protein kinase 1), and non-small cell lung cancer signaling. Activation of antioxidant defense system and immune responses in lungs after paraquat exposure signifies potential induction of pulmonary stresses and diseases. doi:10.1016/j.toxlet.2011.05.741
P2118 LamB, OmpA and OmpC are key altered outer membrane proteins in E. coli response to isopropanol
P2119 Omics for mechanisms of Edwardsiella tarda in resistance to plasma Xuan-Xian Peng ∗ , Xian-Jie Liu, Chang Guo, Xian-Liang Zhao, Hui Li School of Life Sciences, Sun Yat-sen University, Guangzhou 510006, China Purpose: Edwardsiella tarda is known for causing diseases in both humans and fish. Animal plasma is toxic to almost of bacteria, but E. tarda can resist the role. However, information regarding to the mechanisms is not available. In the present study, OMICs approach is used to investigate how the bacterium is free of the plasma toxicity. Methods: 2-DE based proteomics and GC/MS based metabonomics were used to investigate functional proteome and metabolom in resistance to human and fish plasma, respectively. Then, computation biology method was utilized for analysis of pathways and networks. Results of the study: Our results indicate that E. tarda not only resisted to the antimicrobial role of animal plasma, but also survived better with than without plasma. Very interestingly, differently altered proteins and metabolites were detected between human and fish plasma treatments although bacterial OD and live bacterium amounts were similar to the two treatments. WGCNA pathway analysis and pathway analysis indicated that elevation of amino sugar and nucleotide sugar metabolism was found in both plasma, but the resulting pyruvate contributed to pantothenic acid and coenzyme A metabolism alone in the treatment by fish plasma, whereas it entered to one more pathway of peptidoglycan biosynthetic process in the treatment by human plasma. Our finding discloses the molecular mechanisms that E. tarda is free of plasma toxicity. doi:10.1016/j.toxlet.2011.05.743
S. Wang 1,∗ , D. Zhang 1 , X. Lin 2 , H. Li 2 , X. Peng 2 School of Life Sciences, Xiamen University, Xiamen, China, 2 School of Life Sciences, Sun Yat-sen University, Guangzhou, China
1
Purpose: To study the relationship between outer membrane (OM) proteins and isopropanol tolerance, we characterized E. coli K-12 BW25113 OM protein function related to isopropanol at the
Abstracts / Toxicology Letters 205S (2011) S180–S300
haemolyticus as an antigen, VP1061 and VP2850 showed broadly cross-protective ability against V. alginolyticus, A. hydrophila and P. fluorescens in fish and mouse models. In V. alginolyticus as an antigen, the carps immunized with VA0764 and VA1061 had abilities to fight against infections not only caused by V. alginolyticus but also by A. hydrophila and P. fluorescens. These vaccine candidates are importance in against virulent bacterial infections. doi:10.1016/j.toxlet.2011.05.740
P2117 Responses of pulmonary proteins to paraquat in rats Q.X. Li 1,∗ , I.K. Cho 1 , M. Jeong 2 , J.K. Kim 2 , S.S. Hong 2 , K.H. Park 2 1
Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, Honolulu, USA, 2 Crop Life Safety, National Academy of Agricultural Science, Suwon-si, Republic of Korea
sub-proteome level to understand tolerance mechanism of bacteria to disinfectant. Methods: Two dimensional electrophoresis (2-DE) and mass spectrometry (MS) technologies were used to identify differential expression of OMPs of E. coli K-12 BW25113 in response to isopropanol stress. Gene knockout, complementation and overexpression were utilized to investigate the role of altered OM proteins. Results of the study: Up-regulation of LamB, FadL and OmpC, and down-regulation of OmpT, Tsx, OmpA, OmpF were detected. Furthermore, we investigated roles of these altered proteins in resistance to the disinfectant through gene knockout, complementation and overexpression. Our results showed that LamB, OmpA and OmpC played more important role in the resistance than the others. Finally, we characterized the ability of EnvZ/OmpR twocomponent signal transduction system in exposure to disinfectant. These results indicated the importance of OMPs and EnvZ/OmpR two-component signal transduction system in response to isopropanol. doi:10.1016/j.toxlet.2011.05.742
Purpose: Paraquat is among the most widely used herbicides in the world. Paraquat, when ingested, is very toxic to mammals including humans and may cause acute respiratory distress syndrome. The purpose of this study was to investigate the protein profiles in response to paraquat exposure. Methods: Sprague–Dawley male rats were orally administered with paraquat at 5, 10, 25 and 40 mg/kg bw (LD50 = 126 mg/kg) for five times a week for four weeks. Soluble proteins in pulmonary tissues were extracted and analyzed with a shotgun proteomic approach which combined one-dimensional gel electrophoresis, ion trap mass spectrometry and amino acid sequence alignment. Results: A number of 304 and 147 proteins were over- and under-represented, respectively, in the rat lung tissues after exposure to 25 mg/kg or higher of paraquat. The over-represented proteins include C-jun-amino-terminal kinase interacting protein 2, CDKN2A-intercting protein, mitogen-activated protein kinases, macrophage receptor with collagenous structure, complement decay-accelerating factor, stress-70 protein and thioredoxin reductase 3. All the over-and under-represented proteins were subjected to Ingenuity Pathway Analysis to create 25 biological networks and 38 pathways of interacting protein clusters according to the identifiers’ HomoloGene to the ortholog information in the Ingenuity Knowledge Base (Human/Mouse/Rat data). The responded pathways related to pulmonary stress include cardiovascular-cancerrespiratory pathway (e.g., nitric oxide synthase 3), regulation of clathrin-mediated endocytosis (e.g., AP2-associated protein kinase 1), and non-small cell lung cancer signaling. Activation of antioxidant defense system and immune responses in lungs after paraquat exposure signifies potential induction of pulmonary stresses and diseases. doi:10.1016/j.toxlet.2011.05.741
P2118 LamB, OmpA and OmpC are key altered outer membrane proteins in E. coli response to isopropanol
P2119 Omics for mechanisms of Edwardsiella tarda in resistance to plasma Xuan-Xian Peng ∗ , Xian-Jie Liu, Chang Guo, Xian-Liang Zhao, Hui Li School of Life Sciences, Sun Yat-sen University, Guangzhou 510006, China Purpose: Edwardsiella tarda is known for causing diseases in both humans and fish. Animal plasma is toxic to almost of bacteria, but E. tarda can resist the role. However, information regarding to the mechanisms is not available. In the present study, OMICs approach is used to investigate how the bacterium is free of the plasma toxicity. Methods: 2-DE based proteomics and GC/MS based metabonomics were used to investigate functional proteome and metabolom in resistance to human and fish plasma, respectively. Then, computation biology method was utilized for analysis of pathways and networks. Results of the study: Our results indicate that E. tarda not only resisted to the antimicrobial role of animal plasma, but also survived better with than without plasma. Very interestingly, differently altered proteins and metabolites were detected between human and fish plasma treatments although bacterial OD and live bacterium amounts were similar to the two treatments. WGCNA pathway analysis and pathway analysis indicated that elevation of amino sugar and nucleotide sugar metabolism was found in both plasma, but the resulting pyruvate contributed to pantothenic acid and coenzyme A metabolism alone in the treatment by fish plasma, whereas it entered to one more pathway of peptidoglycan biosynthetic process in the treatment by human plasma. Our finding discloses the molecular mechanisms that E. tarda is free of plasma toxicity. doi:10.1016/j.toxlet.2011.05.743
S. Wang 1,∗ , D. Zhang 1 , X. Lin 2 , H. Li 2 , X. Peng 2 School of Life Sciences, Xiamen University, Xiamen, China, 2 School of Life Sciences, Sun Yat-sen University, Guangzhou, China
1
Purpose: To study the relationship between outer membrane (OM) proteins and isopropanol tolerance, we characterized E. coli K-12 BW25113 OM protein function related to isopropanol at the