Primers and probes used in real time PCR analyses.

<p>Primers and probes used in real time PCR analyses.</p

Hematoxylin and eosin (H&E) staining and cross-sectional area (CSA).

<p><b>A, B, C, D</b>) H&E staining for the measurement of CSA of 0.25–4, 5–8 and 9–14 day groups, respectively, <b>E</b>) CSA (μm²) of the masseter muscle fibers, and <b>F</b>) the lesser fiber diameter (μm). Asterisks (**p<0.01) denote significant differences compared with controls. Values are means + SEM.</p

MuRF1, atrogin-1 and LC3b mRNA expression.

<p><b>A</b>) MuRF1 (black bars) and atrogin-1 (grey bars), <b>B</b>) LC3b mRNA expression, according to real-time PCR in the masseter muscle. Asterisks (*p<0.05) denote significant differences compared with controls. Values are means of starting quantities + SEM.</p

Animal treatment.

<p>MV (mechanical ventilation), sepsis (endotoxemia was induced by a continuous infusion of Escherichia coli endotoxin), NMBA (neuromuscular blocking agent, i.e., continuous infusion of rocuronium, 25 mg·h⁻¹), CS (corticosteroid given as bolus doses of hydrocortisone 50 mg, ×3 daily) and ALL (MV+CS+sepsis+NMBA). N: number of animals; BE: base excess; PP: peak pressure.</p

Single muscle fibre size and contractile function.

<p>Cross-sectional area (CSA), specific force (maximal force production normalized to CSA) and maximum unloaded shortening velocity (V₀). Values for day 1 (empty colored bars) and day 5 (hatched colored bars) from mechanically ventilated/sedated/immobilized (MV group) piglets, MV together with neuromuscular blocking agents (NMBA group), MV together with corticosteroid administration (CS group), MV together an endotoxin-induced sepsis (sepsis), and MV together with sepsis, CS and NMBA (ALL group). Data are presented as means ± SEMs. Asterisk denotes a statistically significant difference compared with day 1 (p<0.05).</p

Myosin∶actin ratios.

<p>Values for day 1 (empty colored bars) and day 5 (hatched colored bars) from mechanically ventilated/sedated/immobilized (MV group) piglets, MV together with neuromuscular blocking agents (NMBA group), MV together with corticosteroid administration (CS group), MV together an endotoxin-induced sepsis (sepsis), and MV together with sepsis, CS and NMBA (ALL group). Data are presented as means ± SEMs.</p

Protein and gene expressions.

<p>[A] MyHC isoform composition. [B] Contractile protein content. [C] Total protein over fibre volume. [D] mRNA levels of contractile proteins. Values from sham-operated control piglets (CTL group, white bars) and animals that were mechanically ventilated (MV group, red bars), mechanically ventilated with a neuromuscular blocking agent administration (NMBA group, green bars), mechanically ventilated with corticosteroid administration (CS group, yellow bars), mechanically ventilated with an injection of a endotoxin-induced sepsis (sepsis group, blue bars) and mechanically ventilated with a combination of endotoxin-induced sepsis, CS and NMBA (ALL group, black bars) for five days. Data are presented as means ± SEMs. Asterisk denotes a statistically significant difference compared with CTL (p<0.05).</p

Contractile protein reactive carbonyl derivatives.

<p>[A] CTL (white bars), MV (red bars), NMBA (green bars), CS (yellow bars), sepsis (blue bars) and ALL groups (black bars). [B] Typical western blot showing reactive carbonyl derivates for various piglets. Data are presented as means ± SEMs. Asterisk denotes a statistically significant difference compared with CTL (p<0.05).</p

Animal treatment: CTL (control), MV (mechanical ventilation), sepsis (endotoxemia was induced by a continuous infusion of Escherichia coli endotoxin), NMBA (neuromuscular blocking agent, i.e., continuous infusion of rocuronium, 25 mg·h⁻¹), CS (corticosteroid given as bolus doses of hydrocortisone 50 mg, ×3 daily) and ALL (MV + CS + sepsis + NMBA).

<p>N: number of animals; BE: base excess; PP: peak pressure.</p

Fibre structure and function.

<p>[A] Number of fibres tested. [B] Cross-sectional area (CSA). [C] Maximal force production normalized to CSA (P₀/CSA). [D] Stiffness (E₀). [E] Fraction of strongly attached cross-bridges (α_fs). [F] Apparent rate constant of force redevelopment (k_tr). Values from CTL (white), MV (red bars), NMBA (green), CS (yellow), sepsis (blue) and ALL groups (black) appear. Data are presented as means.</p