Additional file 1: Figure S1. of Influence of weaning methods on the diaphragm after mechanical ventilation in a rat model

Arterial blood gas data in the CMV, PSV and SBT groups. Values are expressed as means ± SD. (JPG 717 kb

FF measurements / CSA data

force frequency measurements <div>cross-sectional areas</div

4 HNE / oxyblot / alpha-spectrin

Western blot

Examples of diaphragmatic leukocyte activity

<p>Diaphragmatic leukocyte activity measured as neutrophils (CD43), ED1+ macrophages (CD68), and ED2+ macrophages (CD163). </p

Comparison of diaphragm contractile force (A) and diaphragm fiber size (B) across the experimental groups.

<p>A) Force-frequency curves for diaphragm B) Fiber cross-sectional area analysis (CSA) of diaphragm sections stained for dystrophin, myosin heavy chain (MHC) I and MHC type IIa proteins. Key: CMV: Mechanical ventilation for 12 hours (n = 7), light grey bar; Ext 12: 12 h of spontaneous breathing after 12 hours of MV (n = 8), dark grey bar; Ext 24: 24 h of spontaneous breathing after 12 hours of MV (n = 8), black bar; CON: unventilated controls (n = 8), white bar. Values are mean ± SD. * significant CMV vs control p<0.05; # significant Ext 12 vs control p<0.05, $ significant CMV vs Ext 24.</p

Western blot results of biomarkers of protein synthesis, proteolysis and oxidative stress in the diaphragm.

<p>A) pAKT/AKT ratio was analyzed as quotient of pAKT versus AKT expression. Representative blots for both AKT and pAKT are displayed above; Con n = 8, CMV n = 6, Ext 12 n = 8, Ext24 n = 8 B) The levels of 4-hydroxynonenal (4- <i>HNE</i>) were analyzed as an indicator of lipid peroxidation via Western blotting. A representative blot for 4-HNE protein conjugates is shown above the graph (Con n = 8, CMV n = 7, Ext 12 n = 8, Ext24 n = 8). C/D) Calpain (C) (Con n = 6, CMV n = 7, Ext 12 n = 7, Ext24 n = 8) and caspase-3 (D) (Con n = 7, CMV n = 7, Ext 12 n = 7, Ext24 n = 8) activations were determined via Western blotting as a ratio for the spectrin breakdown product (120 and 145 kDA band) compared to the 245 kDa band. Representative Western blots are shown above the graphs. CMV 12: Mechanical ventilation for 12 hours, Ext 12: 12 hours of spontaneous breathing after 12 hours of MV, Ext 24: 24 hours of spontaneous breathing after 12 hours of MV, CON: unventilated controls. Values are arbitrary units ± SD. * significant versus Control, # significant versus CMV; IDV integrated density value.</p

Arterial blood gases, blood pressure, Temperature of the animals investigated during the experiment.

<p>CMV 12: Mechanical ventilation for 12 hours, Ext 12: 12 hours of spontaneous breathing after 12 hours of MV, Ext 24: 24 hours of spontaneous breathing after 12 hours of MV, CON: unventilated controls. n.a. not applicable. BP: blood pressure. Body weight determined before sacrifice. + p<0.05 CVM vs Ext 12. * p<0.05 Con vs CMV. Data are mean ± SD.</p

Timeline of the experiments.

<p>CMV: Mechanical ventilation for 12 hours, Ext 12: 12 h of spontaneous breathing after 12 hours of MV, Ext 24: 24 h of spontaneous breathing after 12 hours of MV, CON: unventilated controls. The dotted line indicates the end of the mechanical ventilation period.</p

Pearson correlations (R) between diaphragm specific contractile force and further parameters influenced during mechanical ventilation.

<p>Pearson correlations (R) between diaphragm specific contractile force production (all stimulation frequencies) and 4-HNE concentration, pAkt/Akt ratio, and the cross sectional area of diaphragm type IIb fibers. 4HNE: 4-Hydroxy-non-enal, AKT = Proteinkinase B, CSA: Cross-sectional area.</p

Arterial blood gases, blood pressure and temperature of the animals investigated at the end of the experiment.

<p>CMV 12: Mechanical ventilation for 12 hours, Ext 12: 12 hours of spontaneous breathing after 12 hours of MV, Ext 24: 24 hours of spontaneous breathing after 12 hours of MV, CON: unventilated controls. n.a. not applicable. BP: blood pressure. Body weight determined before sacrifice. + indicates p<0.05 CMV vs Ext 12. Data are mean ± SD.</p