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10.1111/cei.12766 http://scihub22266oqcxt.onion/10.1111/cei.12766 C4872380!4872380!26749379     free     free     free Deprecated: Implicit conversion from float 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Clin+Exp+Immunol 2016 ; 184 (3): 358-67 Nephropedia Template TP {{tp|p=26749379|t=2016. Pneumolysin activates neutrophil extracellular trap formation |pdf=|usr=}}{{26749379}} gab.com Text Pneumolysin activates neutrophil extracellular trap formation JO: Clin Exp Immunol http://www.kidney.de/pget.php?&tw=1&pmid=26749379 #FOAvip The primary objective of the current study was to investigate the potential of the pneumococcal toxin, pneumolysin (Ply), to activate neutrophil extracellular trap (NET) formation in vitro. Isolated human blood neutrophils were exposed to recombinant Ply (5?20 ng ml?1) for 30?90 min at 37°C and NET formation measured using the following procedures to detect extracellular DNA: (i) flow cytometry using Vybrant® DyeCycle? Ruby; (ii) spectrofluorimetry using the fluorophore, Sytox® Orange (5 ?M); and (iii) NanoDrop® technology. These procedures were complemented by fluorescence microscopy using 4?, 6?diamino?2?phenylindole (DAPI) (nuclear stain) in combination with anti?citrullinated histone monoclonal antibodies to visualize nets. Exposure of neutrophils to Ply resulted in relatively rapid (detected within 30?60 min), statistically significant (P < 0·05) dose? and time?related increases in the release of cellular DNA impregnated with both citrullinated histone and myeloperoxidase. Microscopy revealed that NETosis appeared to be restricted to a subpopulation of neutrophils, the numbers of NET?forming cells in the control and Ply?treated systems (10 and 20 ng ml?1) were 4·3 (4·2), 14.3 (9·9) and 16·5 (7·5), respectively (n = 4, P < 0·0001 for comparison of the control with both Ply?treated systems). Ply?induced NETosis occurred in the setting of retention of cell viability, and apparent lack of involvement of reactive oxygen species and Toll?like receptor 4. In conclusion, Ply induces vital NETosis in human neutrophils, a process which may either contribute to host defence or worsen disease severity, depending on the intensity of the inflammatory response during pneumococcal infection. Twit Text FOAVip Pneumolysin activates neutrophil extracellular trap formation ????Clin Exp Immunol http://www.kidney.de/pget.php?&tw=1&pmid=26749379 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=26749379 #FOAvip English Wikipedia <ref>{{Cite pmid|26749379}}</ref> Pneumolysin activates neutrophil extracellular trap formation #MMPMID26749379 G. Nel J; Theron AJ; Durandt C; Tintinger GR; Pool R; Mitchell TJ; Feldman C; Anderson R Clin Exp Immunol 2016[Jun]; 184 (3): 358-67 PMID26749379show ga The primary objective of the current study was to investigate the potential of the pneumococcal toxin, pneumolysin (Ply), to activate neutrophil extracellular trap (NET) formation in vitro. Isolated human blood neutrophils were exposed to recombinant Ply (5?20 ng ml?1) for 30?90 min at 37°C and NET formation measured using the following procedures to detect extracellular DNA: (i) flow cytometry using Vybrant® DyeCycle? Ruby; (ii) spectrofluorimetry using the fluorophore, Sytox® Orange (5 ?M); and (iii) NanoDrop® technology. These procedures were complemented by fluorescence microscopy using 4?, 6?diamino?2?phenylindole (DAPI) (nuclear stain) in combination with anti?citrullinated histone monoclonal antibodies to visualize nets. Exposure of neutrophils to Ply resulted in relatively rapid (detected within 30?60 min), statistically significant (P < 0·05) dose? and time?related increases in the release of cellular DNA impregnated with both citrullinated histone and myeloperoxidase. Microscopy revealed that NETosis appeared to be restricted to a subpopulation of neutrophils, the numbers of NET?forming cells in the control and Ply?treated systems (10 and 20 ng ml?1) were 4·3 (4·2), 14.3 (9·9) and 16·5 (7·5), respectively (n = 4, P < 0·0001 for comparison of the control with both Ply?treated systems). Ply?induced NETosis occurred in the setting of retention of cell viability, and apparent lack of involvement of reactive oxygen species and Toll?like receptor 4. In conclusion, Ply induces vital NETosis in human neutrophils, a process which may either contribute to host defence or worsen disease severity, depending on the intensity of the inflammatory response during pneumococcal infection. äPneumolysin activates neutrophil extracellular trap formation (epmc).pdf DeepDyve Pubget Overpricing