Nski Critical Care 2014, PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/27321907 18:R88 http://ccforum.com/content/18/3/RPage 15 ofcontractile proteins and are independent of diaphragm fiber type. Administration of PEG-SOD, a specific scavenger of superoxide, prevents hyperglycemia-induced diaphragm weakness, restoring both intact diaphragm force generation as well as single fiber diaphragm force generation without lowering glucose levels. Our data identify new mechanisms for hyperglycemia induced diaphragm weakness, and potentially explain how poor glucose control might potentiate the development of diaphragm weakness and prolong the duration of mechanical ventilation during critical illness.Additional filesAdditional file 1: Figure S1. Final diaphragm weight to final animal weight ratios. This is a graph demonstrating the final diaphragm weight to final animal weight ratios in the experimental groups. Additional file 2: Table S1. Fiber type specific data in single permeabilized diaphragm fibers from all experimental groups. This is a table showing the detailed analyses of single fiber experiments based on fiber type as determined by the myosin heavy chain content in individual fibers; included are fiber type specific parameters of maximal force generation per cross sectional area (kPa), cross sectional area, N values (Hill coefficient) and the pCa50. Additional file 3: Figure S2. Effect of insulin treatment on diaphragm specific force generation. This figure demonstrates the effects of insulin treatment in hyperglycemic animals on the diaphragm specific force generation. Abbreviations ANOVA: analysis of variance; Ca50: EPZ-5676 biological activity calcium concentration required for half-maximal activation; COPD: chronic obstructive pulmonary disease; CSA: cross sectional area; dnPEG-SOD: denatured polyethylene glycol conjugated superoxide dismutase; DTT: dithiothreitol; EGTA: ethylene glycol tetraacetic acid; HEPES: 4-(2-hydroxyethyl)-1-piperazine-ethanesulfonic acid; HG: hyperglycemia; HRP: horseradish peroxidase; ICUAW: intensive care unit acquired weakness; JNK: c-Jun N-terminal kinase; kPa: kilopascal; NAC: N-acetyl cysteine; NADH: reduced form of nicotinamide adenine dinucleotide; NADPH oxidase: nicotinamide adenine dinucleotide phosphate-oxidase; pCa: -[log] calcium concentration; PDVF: polyvinylidene fluoride; PEG-SOD: polyethylene glycol conjugated superoxide dismutase; PKR: double-stranded RNA-dependent protein kinase; PMSF: phenylmethanesulfonylfluoride; ROS: reactive oxygen species; SDS: sodium dodecyl sulfate; SEM: standard error of the mean; STZ: streptozotocin. Competing interests Both authors declare that they have no competing interests. Authors’ contributions LAC and GSS conceived and designed the study, supervised the research process, analyzed and interpreted the data, drafted the manuscript and approved the final manuscript. Both authors read and approved the final manuscript. Acknowledgements This work was supported by funds provided by the National Institutes of Health from grants awarded to LAC (R01HL112085, R01HL080609) and to GSS (R01HL113494, R01HL080429, R01HL081525). In addition, the authors would like to thank Drs. Wenyi Wang and Shanshan Zhan for their technical assistance in carrying out some of the studies. Received: 12 December 2013 Accepted: 24 April 2014 Published: 3 MayReferences 1. Latronico N, Bolton CF: Critical illness polyneuropathy and myopathy: a major cause of muscle weakness and paralysis. Lancet Neurol 2011, 10:931?41. 2. Supinski GS, Callahan LA: Diaphragm weakness in m.