The acidosis-induced right shift of the HbO2 dissociation curve is maintained during erythrocyte storage.

BACKGROUND AND OBJECTIVES: In fresh blood, tissue hypoxia increases microcirculatory acidosis, which enhances erythrocyte O(2) unloading and increases the amount of available O(2). Storage of erythrocytes increases the HbO(2) affinity and reduces O(2) unloading. We examined the development of the affinity change during a period of 5 weeks of storage by present blood bank standards, and investigated to what extent acidosis offsets the affinity change. MATERIALS AND METHODS: Blood from volunteer donors was processed and stored as erythrocyte concentrates (EC). At 2-5 day intervals, EC were drawn from the bags and suspended in plasma and crystalloids to an Hb ≈ 10 g/dL. The suspensions were adjusted to give a pH of 7.40, 7.10, 6.80 or 6.30 and equilibrated with different gas mixtures to SO(2) 0, 25, 50, 75 and 100%. Measurements of the PO(2)/SO(2) pairs at each pH were used to calculate the position of the HbO(2) curve and its P(50) value. RESULTS: A significant leftward shift in the HbO(2) curve was established after 1 week of storage; after 2.5 weeks only minor further changes were observed. Acidification right-shifted the HbO(2) curve, after 2.5 weeks of storage the curve at pH 7.10 was similar to that for fresh blood at pH 7.40. Calculations of extractable O(2) showed that the left-shifted HbO(2) curve of stored EC could be advantageous at a low arterial PO(2). CONCLUSIONS: The rightward shift of the HbO(2) curve due to acidosis is well maintained in stored erythrocytes, a moderate pH decrease offsets the storage-induced increased HbO(2) affinity.

Mechanical chest compressions with trapezoidal waveform improve haemodynamics during cardiac arrest.

BACKGROUND: During manual chest compressions for cardiac arrest the waveforms of chest compressions are generally sinusoidal, whereas mechanical chest compression devices can have different waveforms, including trapezoidal. We studied the haemodynamic differences of such waveforms in a porcine model of cardiac arrest. METHODS: Eight domestic pigs (weight 31±3kg) were anaesthetised and instrumented to continuously monitor aortic (AP) and right atrial pressure (RAP), carotid (CF) and cerebral cortical microcirculation blood flow (CCF). Coronary perfusion pressure (CPP) was calculated as the maximal difference between AP and RAP during diastole or decompression phase. After 4 min of electrically induced ventricular fibrillation, mechanical chest compressions were performed with four different waveforms in a factorial design, and in randomized sequence for 3 min each. Resulting differences are presented as mean with 95% confidence intervals. RESULTS: Mean AP and RAP were higher with trapezoid than sinusoid chest compressions, difference 5.7 (0.7, 11) and 6.3 (2.1, 11)mmHg, respectively. Flow measured as CF and CCF was also improved with trapezoidal waveform, difference 14 (2.8, 26)ml/min and 11 (5.6, 17)% of baseline, respectively, with a parallel, non-significant (P=0.08) trend for CPP. Active vs. passive decompression to zero level improved CF, but without even a trend for CPP. CONCLUSION: Trapezoid chest compressions and active decompression to zero level improved blood flow to the brain. The compression waveform is an additional factor to consider when comparing mechanical and manual chest compressions and when comparing different compression devices.

Prearrest administration of low-molecular-weight heparin in porcine cardiac arrest: hemodynamic effects and resuscitability.

OBJECTIVE: Both animal and human studies demonstrate activation of coagulation during cardiac arrest. Prearrest anticoagulation is used routinely in many experimental studies. We studied the hemodynamic effects of prearrest anticoagulation with a low-molecular-weight heparin suitable for clinical use during cardiopulmonary resuscitation in pigs. DESIGN: Randomized and blinded experimental animal study. SETTING: University hospital-affiliated research laboratory. SUBJECTS: Sixteen female domestic pigs. INTERVENTIONS: Three minutes before electrically induced ventricular fibrillation, enoxaparin 1 mg/kg or physiologic saline was blinded and administered intravenously. After 10 mins of untreated ventricular fibrillation, advanced cardiac life support was initiated with continuous mechanical chest compressions and interposed manual ventilation with 100% oxygen. Epinephrine was administered after 2 mins of advanced cardiac life support followed by attempted defibrillation 1 min thereafter. Advanced cardiac life support was continued for 10 mins following international guidelines. Electrocardiogram was recorded continuously and ventricular fibrillation waveform was analyzed (median slope). Animals with return of spontaneous circulation were observed for ten more minutes. Blood specimens were drawn for analysis of coagulation activation (thrombin-antithrombin complex) and drug effect (anti-factor Xa activity). MEASUREMENTS AND MAIN RESULTS: Six of eight (75%) pigs in each group achieved return of spontaneous circulation. Thrombin-antithrombin complex levels were significantly lower in pigs that received enoxaparin. There was no significant difference either in measured hemodynamics between the groups during advanced cardiac life support and after return of spontaneous circulation or in median slope values during ventricular fibrillation. Epinephrine caused a significant decrease in femoral and increase in cerebral cortical blood flow with no difference between the groups. CONCLUSIONS: Prearrest anticoagulation with enoxaparin did not influence either hemodynamics during advanced cardiac life support and after return of spontaneous circulation or the frequency of return of spontaneous circulation in porcine cardiac arrest.

Haemodynamic effects of adrenaline (epinephrine) depend on chest compression quality during cardiopulmonary resuscitation in pigs.

BACKGROUND: Adrenaline (epinephrine) is used during cardiopulmonary resuscitation (CPR) based on animal experiments without supportive clinical data. Clinically CPR was reported recently to have much poorer quality than expected from international guidelines and what is generally done in laboratory experiments. We have studied the haemodynamic effects of adrenaline during CPR with good laboratory quality and with quality simulating clinical findings and the feasibility of monitoring these effects through VF waveform analysis. METHODS AND RESULTS: After 4 min of cardiac arrest, followed by 4 min of basic life support, 14 pigs were randomised to ClinicalCPR (intermittent manual chest compressions, compression-to-ventilation ratio 15:2, compression depth 30-38 mm) or LabCPR (continuous mechanical chest compressions, 12 ventilations/min, compression depth 45 mm). Adrenaline 0.02 mg/kg was administered 30 s thereafter. Plasma adrenaline concentration peaked earlier with LabCPR than with ClinicalCPR, median (range), 90 (30, 150) versus 150 (90, 270) s (p = 0.007), respectively. Coronary perfusion pressure (CPP) and cortical cerebral blood flow (CCBF) increased and femoral blood flow (FBF) decreased after adrenaline during LabCPR (mean differences (95% CI) CPP 17 (6, 29) mmHg (p = 0.01), FBF -5.0 (-8.8, -1.2) ml min(-1) (p = 0.02) and median difference CCBF 12% of baseline (p = 0.04)). There were no significant effects during ClinicalCPR (mean differences (95% CI) CPP 4.7 (-3.2, 13) mmHg (p = 0.2), FBF -0.2 (-4.6, 4.2) ml min(-1)(p = 0.9) and CCBF 3.6 (-1.8, 9.0)% of baseline (p = 0.15)). Slope VF waveform analysis reflected changes in CPP. CONCLUSION: Adrenaline improved haemodynamics during laboratory quality CPR in pigs, but not with quality simulating clinically reported CPR performance.

Slow diffusion of K+ in the T tubules of rat cardiomyocytes.

Cardiomyocyte contractility is regulated by the extracellular K(+) concentration ([K(+)](o)). Potassium dynamics in the T tubules during the excitation-contraction cycle depends on the diffusion rate of K(+), but this rate is not known. Detubulation of rat cardiomyocytes was induced by osmotic shock using formamide, which separated the surface membrane from the T tubules. Changes in current and membrane potential in voltage-clamped (-80 mV) and current-clamped control and detubulated cardiomyocytes were compared during rapid switches between 5.4 and 8.1 mM [K(+)](o), and the results were simulated in a mathematical model. In the voltage-clamp experiments, the current changed significantly slower in control than in detubulated cardiomyocytes during the switch from 5.4 to 8.1 mM [K(+)](o), as indicated by the times to achieve 25, 50, 90, and 95% of the new steady-state current [control (ms) t(25) = 98 +/- 12, t(50) = 206 +/- 20, t(90) = 570 +/- 72, t(95) = 666 +/- 92; detubulated t(25) = 61 +/- 11, t(50) = 142 +/- 17, t(90) = 352 +/- 52, t(95) = 420 +/- 69]. These time points were not significantly different either during the 8.1 to 5.4 mM [K(+)](o) switch or in current-clamped cardiomyocytes switching from 5.4 to 8.1 mM [K(+)](o). Mathematical simulation of the difference current between control and detubulated cardiomyocytes gave a t-tubular diffusion rate for K(+) of approximately 85 mum(2)/s. We conclude that the diffusion of K(+) in the T tubules is so slow that they constitute a functional compartment. This might play a key role in local regulation of the action potential, and thus in the regulation of cardiomyocyte contractility.

T-tubule disorganization and reduced synchrony of Ca2+ release in murine cardiomyocytes following myocardial infarction.

In cardiac myocytes, initiation of excitation-contraction coupling is highly localized near the T-tubule network. Myocytes with a dense T-tubule network exhibit rapid and homogeneous sarcoplasmic reticulum (SR) Ca(2+) release throughout the cell. We examined whether progressive changes in T-tubule organization and Ca(2+) release synchrony occur in a murine model of congestive heart failure (CHF). Myocardial infarction (MI) was induced by ligation of the left coronary artery, and CHF was diagnosed by echocardiography (left atrial diameter >2.0 mm). CHF mice were killed at 1 or 3 weeks following MI (1-week CHF, 3-week CHF) and cardiomyocytes were isolated from viable regions of the septum, excluding the MI border zone. Septal myocytes from SHAM-operated mice served as controls. T-tubules were visualized by confocal microscopy in cells stained with di-8-ANEPPS. SHAM cells exhibited a regular striated T-tubule pattern. However, 1-week CHF cells showed slightly disorganized T-tubule structure, and more profound disorganization occurred in 3-week CHF with irregular gaps between adjacent T-tubules. Line-scan images of Ca(2+) transients (fluo-4 AM, 1 Hz) showed that regions of delayed Ca(2+) release occurred at these gaps. Three-week CHF cells exhibited an increased number of delayed release regions, and increased overall dyssynchrony of Ca(2+) release. A common pattern of Ca(2+) release in 3-week CHF was maintained between consecutive transients, and was not altered by forskolin application. Thus, progressive T-tubule disorganization during CHF promotes dyssynchrony of SR Ca(2+) release which may contribute to the slowing of SR Ca(2+) release in this condition.