Bronchoalveolar lavage fluid (BALF) cytokine, chemokine and protein concentrations in mice after sham procedure or double hit (DH) in C5^+/+ (wt) mice (black bars) and C5^-/- mice (grey bars) and MPO activity in lung tissue.

<p>Increased IL-6 BALF concentration in C5^-/- mice after DH compared to sham and compared to DH in wt mice. <b>B</b>. Increased MCP-1 levels after DH in absence of C5 compared to sham and compared to DH in wt mice. <b>C</b>. Increased G-CSF BALF concentration after DH in presence and absence of C5. Further increase of G-CSF concentration after DH in absence of C5 compared to wt. <b>D</b>. Increased KC levels in wt mice after DH. <b>E</b>. Increased protein concentrations in bronchoalveolar lavage fluids (BALF) after DH in both wt (black bars) and in absence of C5 (grey bars). Further increase in BALF protein concentrations in C5^-/- after DH compared to DH in wt mice. <b>F</b>. MPO activity in lung tissue is increased after DH compared to sham in both wt and in absence of C5 and further increased after DH in absence of C5. For each bar, n = 7 separate mice. * p<0.05, n.s. = not significant.</p

Cytokines and chemokines in lung tissue after sham procedure or double hit (DH) in wild type (wt) mice (black bars) or in absence of C5 (C5^-/-) mice (grey bars). Systemic effects of the absence of complement C5 in double hit (DH).

<p><b>A</b>. Increased IL-6 levels in lung tissue of wt and in C5^-/- mice after DH compared to sham. <b>B</b>. Increased MCP-1 levels in lung tissue of wt and in C5^-/- after DH compared to sham. <b>C</b>. Increased G-CSF levels in lung tissue of wt and in C5^-/- mice after DH compared to sham. <b>D</b>. Increased KC levels in lung tissue of wt mice. <b>E</b>. Increased KC plasma concentration in wt mice after DH; decreased KC plasma concentrations after DH in C5^-/- compared wt mice. <b>F</b>. Increased MCP-1 plasma concentrations in wt mice after DH, decrease of MCP-1 after DH in absence of C5. For each bar, n = 7 separate mice. * p<0.05, n.s. = not significant.</p

Table_3_Successful Resuscitation in a Model of Asphyxia and Hemorrhage to Test Different Volume Resuscitation Strategies. A Study in Newborn Piglets After Transition.pdf

<p>Background: Evidence for recommendations on the use of volume expansion during cardiopulmonary resuscitation in newborn infants is limited.</p><p>Objectives: To develop a newborn piglet model with asphyxia, hemorrhage, and cardiac arrest to test different volume resuscitation on return of spontaneous circulation (ROSC). We hypothesized that immediate red cell transfusion reduces time to ROSC as compared to the use of an isotonic crystalloid fluid.</p><p>Methods: Forty-four anaesthetized and intubated newborn piglets [age 32 h (12–44 h), weight 1,220 g (1,060–1,495g), Median (IQR)] were exposed to hypoxia and blood loss until asystole occurred. At this point they were randomized into two groups: (1) Crystalloid group: receiving isotonic sodium chloride (n = 22). (2) Early transfusion group: receiving blood transfusion (n = 22). In all other ways the piglets were resuscitated according to ILCOR 2015 guidelines [including respiratory support, chest compressions (CC) and epinephrine use]. One hour after ROSC piglets from the crystalloid group were randomized in two sub-groups: late blood transfusion and infusion of isotonic sodium chloride to investigate the effects of a late transfusion on hemodynamic parameters.</p><p>Results: All animals achieved ROSC. Comparing the crystalloid to early blood transfusion group blood loss was 30.7 ml/kg (22.3–39.6 ml/kg) vs. 34.6 ml/kg (25.2–44.7 ml/kg), Median (IQR). Eleven subjects did not receive volume expansion as ROSC occurred rapidly. Thirty-three animals received volume expansion (16 vs. 17 in the crystalloid vs. early transfusion group). 14.1% vs. 10.5% of previously extracted blood volume in the crystalloid vs. early transfusion group was infused before ROSC. There was no significant difference in time to ROSC between groups [crystalloid group: 164 s (129–198 s), early transfusion group: 163 s (162–199 s), Median (IQR)] with no difference in epinephrine use.</p><p>Conclusions: Early blood transfusion compared to crystalloid did not reduce time to ROSC, although our model included only a moderate degree of hemorrhage and ROSC occurred early in 11 subjects before any volume resuscitation occurred.</p

Early structural changes of the heart after experimental polytrauma and hemorrhagic shock

<div><p>Evidence is emerging that systemic inflammation after trauma drives structural and functional impairment of cardiomyocytes and leads to cardiac dysfunction, thus worsening the outcome of polytrauma patients. This study investigates the structural and molecular changes in heart tissue 4 h after multiple injuries with additional hemorrhagic shock using a clinically relevant rodent model of polytrauma. We determined mediators of systemic inflammation (keratinocyte chemoattractant, macrophage chemotactic protein 1), activated complement component C3a and cardiac troponin I in plasma and assessed histological specimen of the mouse heart via standard histomorphology and immunohistochemistry for cellular and subcellular damage and ongoing apoptosis. Further we investigated spatial and quantitative changes of connexin 43 by immunohistochemistry and western blotting. Our results show significantly increased plasma levels of both keratinocyte chemoattractant and cardiac troponin I 4 h after polytrauma and 2 h after induction of hypovolemia. Although we could not detect any morphological changes, immunohistochemical evaluation showed increased level of tissue high-mobility group box 1, which is both a damage-associated molecule and actively released as a danger response signal. Additionally, there was marked lateralization of the cardiac gap-junction protein connexin 43 following combined polytrauma and hemorrhagic shock. These results demonstrate a molecular manifestation of remote injury of cardiac muscle cells in the early phase after polytrauma and hemorrhagic shock with marked disruption of the cardiac gap junction. This disruption of an important component of the electrical conduction system of the heart may lead to arrhythmia and consequently to cardiac dysfunction.</p></div

Experimental blunt chest trauma-induced myocardial inflammation and alteration of gap-junction protein connexin 43

<div><p>Objective</p><p>Severe blunt chest trauma in humans is associated with high mortality rates. Whereas lung tissue damage and lung inflammation after blunt chest trauma have extensively been investigated, the traumatic and posttraumatic effects on the heart remain poorly understood. Therefore, the purpose of this study was to define cardiac injury patterns in an experimental blunt chest trauma model in rats.</p><p>Methods</p><p>Experimental blunt chest trauma was induced by a blast wave in rats, with subsequent analysis of its effects on the heart. The animals were subjected either to a sham or trauma procedure. Systemic markers for cardiac injury were determined after 24 h and 5 days. Postmortem analysis of heart tissue addressed structural injury and inflammation 24 h and 5 days after trauma.</p><p>Results</p><p>Plasma levels of extracellular histones were elevated 24 h and 5 days after blunt chest trauma compared to sham-treated animals. In the heart, up-regulation of interleukin-1β 24 h after trauma and increased myeloperoxidase activity 24 h and 5 days after trauma were accompanied by reduced complement C5a receptor-1 expression 24 h after trauma. Histological analysis revealed extravasation of erythrocytes and immunohistochemical analysis alteration of the pattern of the gap-junction protein connexin 43. Furthermore, a slight reduction of α-actinin and desmin expression in cardiac tissue was found after trauma together with a minor increase in sarcoplasmatic/endoplasmatic reticlulum calcium-ATPase (SERCA) expression.</p><p>Conclusions</p><p>The clinically highly relevant rat model of blast wave-induced blunt chest trauma is associated with cardiac inflammation and structural alterations in cardiac tissue.</p></div

Histological and humoral markers of cardiac damage.

<p>(A) Plasma levels of cardiac troponin I (cTnI) as a marker of cardiac damage from animals 4 h after the insult of polytrauma and hemorrhagic shock (PTHS; n = 8) and native controls (CTRL, n = 4). Results are presented as amount of protein per total protein in plasma to unmask diluting effects from volume resuscitation. (B) Representative images of immunohistochemical (IHC) assessment of tissue cleaved caspase 3. Magnification: 100x (bar: 50 μm). (C) Representative hematoxylin and eosin stained sections of cardiac tissue, showing no signs of marked histomorphological changes. Magnifications: 100x (upper images, bar: 50 μm) and 200x (lower images, bar: 100 μm) for each group. (D) Representative images and densitometric analysis of IHC preparations of tissue high-mobility group box 1 (HMGB1) showing increased signal in samples from PTHS animals. Magnification: 100x (bar: 50 μm). For histological evaluation: n = 5 (PTHS); n = 5 (CTRL). DSR: density sum red. Experimental means were compared for statistical significance using the unpaired t-test (D) and Mann-Whitney rank sum test (A). *: p<0.05.</p

Plasma levels of inflammatory mediators and hemoglobin.

<p>Levels of (A) haemoglobin (Hb) in blood and (B) macrophage chemotactic protein 1 (MCP-1), (C) keratinocyte chemoattractant (KC) and (D) complement component C3a in plasma of animals 4 h after infliction of polytrauma and hemorrhagic shock (PTHS; n = 7 for Hb, n = 8 for MCP-1, KC and C3a) and native control animals (CTRL; n = 8 for Hb, n = 4 for MCP-1, n = 5 for C3a and KC). Results (B, C, D) are presented as amount of protein per total protein in plasma to unmask diluting effects from volume resuscitation. For statistical comparison of experimental means, unpaired t-tests (A, C, D) and Mann-Whitney rank sum test (B) were performed. *: p<0.05.</p

Local inflammation in cardiac injury after blunt chest trauma.

<p>A. Increased myeloperoxidase (MOP) activity in left ventricular cardiac tissue 24 h and 5 d after blunt chest trauma compared to sham procedure. B. Elevation of proinflammatory cardiodepressive cytokine IL-1β expression in left ventricular homogenates 24 h after blunt chest trauma compared to sham procedure as assessed by qRT-PCR. C. Representative western blot for C5aR1 of left ventricular tissue homogenates. Densitometry revealed diminished C5aR1 protein expression in left ventricular homogenates 24 h after blunt chest trauma compared to sham procedure. D. C5aR expression in left ventricular homogenates 24 h and 5 d after blunt chest trauma and after sham procedure as assessed by qRT-PCR. p<0.05; *differences were significant to sham procedure, For all frames n = 8 for each bar.</p

Systemic and local effects of blunt chest trauma.

<p>Apparence of extracellular histones in plasma 24 h and 5 d after blunt chest trauma or sham, * differences to sham procedure were significant, p<0.05; n = 4 for each bar. B. Representative H.E staining of left ventricles 24 h or 5 d after blunt chest trauma or sham procedure as indicated.</p

Structural alterations in the heart after blunt chest trauma.

<p>Alteration of gap junctional protein connexin 43 (Cx43) after blunt chest trauma in the heart. A. Representative distribution of Cx43 in cardiac tissue after sham procedure or 24 h or 5d after blunt chest trauma as indicated. B. Representative western blot for Cx43 of left ventricular tissue homogenates. Densitometry revealed no significant increase in protein expression in left ventricular homogenates 24 h and 5 d after blunt chest trauma compared to sham procedure. For all frames n = 4 for each bar.</p