Cardioplegic strategies to protect the hypertrophic heart during cardiac surgery.

Cardioplegic arrest and cardiopulmonary bypass are key triggers of myocardial injury during aortic valve surgery. Cardioplegic ischaemic arrest is associated with disruption to metabolic and ionic homeostasis in cardiomyocytes. These changes predispose the heart to reperfusion injury caused by elevated intracellular reactive oxygen species and calcium. Cardiopulmonary bypass is associated with an inflammatory response that can generate systemic oxidative stress which, in turn, provokes further damage to the heart. Techniques of myocardial protection are routinely applied to all hearts, irrespective of their pathology, although different cardiomypathies respond differently to ischaemia and reperfusion injury. In particular, the efficacy of cardioprotective interventions used to protect the hypertrophic heart in patients with aortic valve disease remains controversial. This review will describe key cellular changes in hypertrophy, response to ischaemia and reperfusion and cardioplegic arrest and highlight the importance of optimising cardioprotective strategies to suit hypertrophic hearts.

Acute regional changes in myocardial strain may predict ventricular remodelling after myocardial infarction in a large animal model.

To identify predictors of left ventricular remodelling (LVR) post-myocardial infarction (MI) and related molecular signatures, a porcine model of closed-chest balloon MI was used along with serial cardiac magnetic resonance imaging (CMRI) up to 5-6 weeks post-MI. Changes in myocardial strain and strain rates were derived from CMRI data. Tissue proteomics was compared between infarcted and non-infarcted territories. Peak values of left ventricular (LV) apical circumferential strain (ACS) changed over time together with peak global circumferential strain (GCS) while peak GLS epicardial strains or strain rates did not change over time. Early LVR post-MI enhanced abundance of 39 proteins in infarcted LV territories, 21 of which correlated with LV equatorial circumferential strain rate. The strongest associations were observed for D-3-phosphoglycerate dehydrogenase (D-3PGDH), cysteine and glycine-rich protein-2, and secreted frizzled-related protein 1 (sFRP1). This study shows that early changes in regional peak ACS persist at 5-6 weeks post-MI, when early LVR is observed along with increased tissue levels of D-3PGDH and sFRP1. More studies are needed to ascertain if the observed increase in tissue levels of D-3PGDH and sFRP1 might be casually involved in the pathogenesis of adverse LV remodelling.

The importance of myocardial amino acids during ischemia and reperfusion in dilated left ventricle of patients with degenerative mitral valve disease.

Taurine, glutamine, glutamate, aspartate, and alanine are the most abundant intracellular free amino acids in human heart. The myocardial concentration of these amino acids changes during ischemia and reperfusion due to alterations in metabolic and ionic homeostasis. We hypothesized that dilated left ventricle secondary to mitral valve disease has different levels of amino acids compared to the right ventricle and that such differences determine the extent of amino acids' changes during ischemia and reperfusion. Myocardial concentration of amino acids was measured in biopsies collected from left and right ventricles before cardioplegic arrest (Custodiol HTK) and 10 min after reperfusion in patients undergoing mitral valve surgery. The dilated left ventricle had markedly higher (P < 0.05) concentrations (nmol/mg wet weight) of taurine (17.0 +/- 1.5 vs. 10.9 +/- 1.5), glutamine (20.5 +/- 2.4 vs. 12.1 +/- 1.2), and glutamate (18.3 +/- 2.2 vs. 11.4 +/- 1.5) when compared to right ventricle. There were no differences in the basal levels of alanine or aspartate. Upon reperfusion, a significant (P < 0.05) fall in taurine and glutamine was seen only in the left ventricle. These changes are likely to be due to transport (taurine) and/or metabolism (glutamine). There was a marked increase in the alanine to glutamate ratio in both ventricles indicative of ischemic stress which was confirmed by global release of lactate during reperfusion. This study shows that in contrast to the right ventricle, the dilated left ventricle had remodeled to accumulate amino acids which are used during ischemia and reperfusion. Whether these changes reflect differences in degree of cardioplegic protection between the two ventricles remain to be investigated.

Participation of opioid receptors in the cytoprotective effect of chronic normobaric hypoxia.

We studied the role of the delta, micro, and kappa opioid receptor (OR) subtypes in the cardioprotective effect of chronic continuous normobaric hypoxia (CNH) in the model of acute anoxia-reoxygenation of isolated cardiomyocytes. Adaptation of rats to CNH was performed by their exposure to atmosphere containing 12 % of O(2) for 21 days. Anoxia-reoxygenation of cardiomyocytes isolated from normoxic control rats caused the death of 51 % of cells and lactate dehydrogenase (LDH) release. Adaptation of rats to CNH resulted in the anoxia/reoxygenation-induced cardiomyocyte death of only 38 %, and reduced the LDH release by 25 %. Pre-incubation of the cells with either the non-selective OR (opioid receptor) blocker naloxone (300 nM/l), the delta OR antagonist TIPP(psi) (30 nM/l), the selective delta(2) OR antagonist naltriben (1 nM/l) or the micro OR antagonist CTAP (100 nM/l) for 25 minutes before anoxia abolished the reduction of cell death and LDH release afforded by CNH. The antagonist of delta(1) OR BNTX (1 nM/l) or the kappa OR antagonist nor-binaltorphimine (3 nM/l) did not influence the cytoprotective effects of CNH. Taken together, the cytoprotective effect of CNH is associated with the activation of the delta(2) and micro OR localized on cardiomyocytes.

The cardiac proteome in patients with congenital ventricular septal defect: A comparative study between right atria and right ventricles.

Right ventricle (RV) remodelling occurs in neonatal patients born with ventricular septal defect (VSD). The presence of a defect between the two ventricles allows for shunting of blood from the left to right side. The resulting RV hypertrophy leads to molecular remodelling which has thus far been largely investigated using right atrial (RA) tissue. In this study we used proteomic and phosphoproteomic analysis in order to determine any difference between the proteomes for RA and RV. Samples were therefore taken from the RA and RV of five infants (0.34 ±â€¯0.05 years, mean ±â€¯SEM) with VSD who were undergoing cardiac surgery to repair the defect. Significant differences in protein expression between RV and RA were seen. 150 protein accession numbers were identified which were significantly lower in the atria, whereas none were significantly higher in the atria compared to the ventricle. 19 phosphorylation sites (representing 19 phosphoproteins) were also lower in RA. This work has identified differences in the proteome between RA and RV which reflect differences in contractile activity and metabolism. As such, caution should be used when drawing conclusions based on analysis of the RA and extrapolating to the hypertrophied RV. SIGNIFICANCE: RV hypertrophy occurs in neonatal patients born with VSD. Very little is known about how the atria responds to RV hypertrophy, especially at the protein level. Access to tissue from age-matched groups of patients is very rare, and we are in the unique position of being able to get tissue from both the atria and ventricle during reparative surgery of these infants. Our findings will be beneficial to future research into heart chamber malformations in congenital heart defects.

Effect of Chronic Continuous Normobaric Hypoxia on Functional State of Cardiac Mitochondria and Tolerance of Isolated Rat Heart to Ischemia and Reperfusion: Role of µ and delta2 Opioid Receptors.

Chronic continuous normobaric hypoxia (CNH) increases cardiac tolerance to ischemia/reperfusion injury in vivo and this effect is mediated via µ and delta2 opioid receptors (ORs) activation. CNH has also been shown to be cardioprotective in isolated rat heart. In this study, we hypothesize that this cardioprotective effect of CNH is mediated by activation of µ and delta2 ORs and preservation of mitochondrial function. Hearts from rats adapted to CNH (12 % oxygen) for 3 weeks were extracted, perfused in the Langendorff mode and subjected to 45 min of global ischemia and 30 min of reperfusion. Intervention groups were pretreated for 10 min with antagonists for different OR types: naloxone (300 nmol/l), the selective delta OR antagonist TIPP(psi) (30 nmol/l), the selective delta1 OR antagonist BNTX (1 nmol/l), the selective delta2 OR antagonist naltriben (1 nmol/l), the selective peptide µ OR antagonist CTAP (100 nmol/l) and the selective delta OR antagonist nor-binaltorphimine (3 nmol/l). Creatine kinase activity in coronary effluent and cardiac contractile function were monitored to assess cardiac injury and functional impairment. Additionally, cardiac tissue was collected to measure ATP and to isolate mitochondria to measure respiration rate and calcium retention capacity. Adaptation to CNH decreased myocardial creatine kinase release during reperfusion and improved the postischemic recovery of contractile function. Additionally, CNH improved mitochondrial state 3 and uncoupled respiration rates, ADP/O, mitochondrial transmembrane potential and calcium retention capacity and myocardial ATP level during reperfusion compared to the normoxic group. These protective effects were completely abolished by naloxone, TIPP(psi), naltriben, CTAP but not BNTX or nor-binaltorphimine. These results suggest that cardioprotection associated with adaptation to CNH is mediated by µ and delta2 opioid receptors activation and preservation of mitochondrial function.

Inflammatory response and cardioprotection during open-heart surgery: the importance of anaesthetics.

Open-heart surgery triggers an inflammatory response that is largely the result of surgical trauma, cardiopulmonary bypass, and organ reperfusion injury (e.g. heart). The heart sustains injury triggered by ischaemia and reperfusion and also as a result of the effects of systemic inflammatory mediators. In addition, the heart itself is a source of inflammatory mediators and reactive oxygen species that are likely to contribute to the impairment of cardiac pump function. Formulating strategies to protect the heart during open heart surgery by attenuating reperfusion injury and systemic inflammatory response is essential to reduce morbidity. Although many anaesthetic drugs have cardioprotective actions, the diversity of the proposed mechanisms for protection (e.g. attenuating Ca(2+) overload, anti-inflammatory and antioxidant effects, pre- and post-conditioning-like protection) may have contributed to the slow adoption of anaesthetics as cardioprotective agents during open heart surgery. Clinical trials have suggested at least some cardioprotective effects of volatile anaesthetics. Whether these benefits are relevant in terms of morbidity and mortality is unclear and needs further investigation. This review describes the main mediators of myocardial injury during open heart surgery, explores available evidence of anaesthetics induced cardioprotection and addresses the efforts made to translate bench work into clinical practice.

Hypoxic Pulmonary Vasoconstriction in Humans: Tale or Myth.

Hypoxic Pulmonary vasoconstriction (HPV) describes the physiological adaptive process of lungs to preserves systemic oxygenation. It has clinical implications in the development of pulmonary hypertension which impacts on outcomes of patients undergoing cardiothoracic surgery. This review examines both acute and chronic hypoxic vasoconstriction focusing on the distinct clinical implications and highlights the role of calcium and mitochondria in acute versus the role of reactive oxygen species and Rho GTPases in chronic HPV. Furthermore it identifies gaps of knowledge and need for further research in humans to clearly define this phenomenon and the underlying mechanism.

Cellular and molecular basis of RV hypertrophy in congenital heart disease.

RV hypertrophy (RVH) is one of the triggers of RV failure in congenital heart disease (CHD). Therefore, improving our understanding of the cellular and molecular basis of this pathology will help in developing strategic therapeutic interventions to enhance patient benefit in the future. This review describes the potential mechanisms that underlie the transition from RVH to RV failure. In particular, it addresses structural and functional remodelling that encompass contractile dysfunction, metabolic changes, shifts in gene expression and extracellular matrix remodelling. Both ischaemic stress and reactive oxygen species production are implicated in triggering these changes and will be discussed. Finally, RV remodelling in response to various CHDs as well as the potential role of biomarkers will be addressed.

Effect of ischaemia and reperfusion on the intracellular concentration of taurine and glutamine in the hearts of patients undergoing coronary artery surgery.

Taurine and glutamine are the most abundant intracellular free amino acids in mammalian hearts where changes in their intracellular concentrations are likely to influence a number of cellular activities. In this study we investigated the effects of ischaemia and reperfusion on the intracellular concentrations of taurine and glutamine in the hearts of patients undergoing coronary artery bypass surgery using cold crystalloid or cold blood cardioplegic solutions. Ischaemic arrest (30 min), using cold crystalloid cardioplegic solution (n = 19), decreased the intracellular concentrations (micromol/g wet weight) of taurine (from 9.8 +/- 0.8 to 7.7 +/- 0.7, P < 0.05) and glutamine (8.7 +/- 0.5 to 7.2 +/- 0.6). After 20 min of normothermic reperfusion the fall in taurine and glutamine was maintained (7.5 +/- 0.5 and 7.4 +/- 0.7 for taurine and glutamine respectively). Myocardial ischaemic arrest with cold blood cardioplegic solution (n = 16) did not cause a significant fall in tissue taurine or glutamine. However, on reperfusion there was a marked fall in the intracellular concentrations of taurine (9.4 +/- 0.5 to 6.5 +/- 0.7) and glutamine (8.0 +/- 0.7 to 5.8 +/- 0.4). The fall in amino acids was associated with a fall in ATP and a rise in tissue lactate. This work demonstrates that irrespective of the cardioplegic solution used to arrest the heart, there is a marked fall in tissue taurine and glutamine which may influence the extent of recovery following surgery. The fall in taurine is largely due to efflux whereas changes in glutamine are due to both transport and metabolism. Ischaemia, hypothermia and changes in the transmembrane concentration gradients are the likely factors responsible for the changes in tissue amino acids.

Age-dependent and hypoxia-related differences in myocardial protection during pediatric open heart surgery.

BACKGROUND: Current cardioplegic protection techniques used in pediatric cardiac surgery do not take into consideration age and cyanotic differences. The aim of the present work was to address this question by monitoring clinical outcome, myocardial metabolism, and reperfusion injury in pediatric patients protected by cold-crystalloid cardioplegia. METHODS AND RESULTS: Fifty-eight patients (31 children and 27 infants) with or without hypoxic stress (cyanosis) undergoing open heart surgery with cold-crystalloid cardioplegia were included in the study. Clinical outcome measures assessed included inotropic and ventilatory support, intensive care, and hospital stay. Ischemia-induced changes in metabolism (adenine nucleotides, purines, lactate, and amino acids) were determined in ventricular biopsies collected at the beginning and end of ischemic time (cross-clamp time). Reperfusion injury was assessed by measuring postoperative serial release of troponin I. Evidence was observed of ischemic stress during cardioplegic arrest in children and infants as shown by significant changes in cellular metabolites. Compared with infants, children had significantly less reperfusion injury and better clinical outcome, and these factors were related to duration of ischemic time. Cyanosis did not influence outcome in infants, but cyanotic children showed worse reperfusion injury and clinical outcome than acyanotic children. CONCLUSIONS: Extent of myocardial protection with cold-crystalloid cardioplegia in pediatric open heart surgery is dependent on age and degree of cyanosis.

Mitochondria: a target for myocardial protection.

The ischemic heart requires reperfusion using clinical interventions, such as coronary artery bypass graft surgery, in order to recover. Despite recent developments in myocardial protection techniques, reperfusion damage still occurs, and significant morbidity remains a problem. Therefore, the search continues for techniques that will limit myocardial damage and that will enhance recovery upon reperfusion. Mitochondria are known to be intimately involved in the processes that lead to cell death following reperfusion, in both necrotic and apoptotic forms of cell death, and so are potential targets for protective intervention. In this review, we consider several aspects of mitochondrial function that we believe to be possible targets for myocardial protection; namely, mitochondrial Ca(2+) transport, the permeability transition pore, and improved mitochondrial substrate supply. We discuss work by ourselves and others in these areas, and also consider the recently proposed role of mitochondrial ATP-dependent K(+) channels in mediating myocardial protection by ischemic preconditioning. Finally, we describe use of cardioplegic solutions in the clinical setting, and discuss how improved understanding of the aspects of mitochondrial function summarised above may lead to better protective strategies in the future.

Changes in the principal free intracellular amino acids in the Langendorff perfused guinea pig heart during arrest with calcium-free or high potassium media.

OBJECTIVE: Many amino acids have been shown to be cotransported with Na+ in a variety of tissues but evidence for an Na+ dependent efflux in the heart is restricted to taurine. The aim of this study was to determine the intracellular levels of the principal amino acids, alanine, glutamine, glutamate, and aspartate, when the heart was beating at normal or low temperature, during exposure to strophanthidin, and when the heart was arrested with either Ca2+,Mg(2+)-free Tyrode solution (a condition known to raise intracellular Na+) or with a high K+ Tyrode solution. METHODS: Guinea pig hearts, mounted on a Langendorff apparatus, were perfused with a variety of media and the level of amino acids in the ventricles determined using high performance liquid chromatography. RESULTS: During perfusion with normal Tyrode solution a time dependent fall in tissue glutamine, glutamate, and aspartate, but not alanine, was observed. Exposure of the beating heart to strophanthidin (0.2 mM) or cooling to 20 degrees C, which should induce a similar increase in [Na+]i but have opposite effects on metabolism, had different effects on tissue amino acids. Cooling did not affect the level of amino acids whereas strophanthidin produced an effect consistent with an activation of a glutamate-alanine aminotransferase. Arresting the heart with Ca2+,Mg(2+)-free Tyrode solution provoked a marked fall in all amino acids which was associated with their appearance in the effluent. This loss of the amino acids was antagonised by conditions known to reduce the rise in intracellular Na+ concentration and increased by inhibition of the Na+ pump. Comparison of the beating heart with the heart arrested with high K+ Tyrode solution showed no difference in the levels of glutamate or aspartate, with a small fall in alanine and taurine and raised glutamine. CONCLUSIONS: The data from beating or arrested hearts are consistent with the presence of amino acid/Na+ cotransport systems but the levels of amino acids free in the sarcoplasm depend on an interaction between amino acid transport driven by the changes in the Na+ gradient and the effects of metabolism.

Changes in intracellular concentration of amino acids in human saphenous vein during preparation for coronary artery bypass grafting.

OBJECTIVES: Earlier research on human skeletal and cardiac muscle has shown a fall in the intracellular free amino acid pool, particularly the non-essential amino acids, during surgery. Surgical preparation of the human saphenous vein for coronary artery by pass grafting has been shown to provoke metabolic damage and may therefore induce changes in cellular amino acids which may have long-term implications for the performance of the grafted vein. This work investigates the hypothesis that surgical preparation of saphenous vein induces changes in amino acids. METHODS: Changes in the intracellular free amino acid pool were monitored, using high-performance liquid chromatography (HPLC), in both freshly isolated and surgically prepared human saphenous veins during coronary artery bypass grafting. In order to asses the extent of metabolic damage incurred by surgical preparation, adenosine triphosphate (ATP) levels were also measured. RESULTS: A substantial fall in intracellular free amino acid was associated with surgical preparation of human saphenous vein for coronary artery bypass grafting (190.8 +/- 25 to 106 +/- 18 mumol.g-1 protein, n = 10, P < 0.05). Although the fall was seen in most amino acids detected, it was marked and statistically significant (P < 0.05) for taurine (30.6 +/- 3.1 to 9.0 +/- 2.2 mumol.g-1 protein), glutamate (43.6 +/- 5.6 to 18.2 +/- 4.5 mumol.g-1 protein), glutamine (12.8 +/- 1.6 to 3.3 +/- 1.2 mumol.g-1 protein) and asparagine (9.6 +/- 1.6 to 4.1 +/- 1.0 mumol.g-1 protein). The fall in tissue taurine which would be largely due to transport showed a strong positive correlation with the fall seen in glutamate, glutamine and asparagine. ATP levels significantly decreased as a result of surgical preparation (279 +/- 44 to 96 +/- 21 nmol.g-1 wet weight, P < 0.05). The fall in ATP was accompanied by an increase in alanine/glutamate ratio which may reflect increased glutamate-alanine transaminase activity and therefore metabolic changes. CONCLUSIONS: The metabolic damage associated with surgical preparation of human saphenous vein for coronary artery bypass grafting is accompanied by a fall in the intracellular free amino acid pool which is marked and statistically significant for glutamate, taurine, glutamine and asparagine. The fall is likely to be due to transport metabolism may also contribute. This may have important implications for the functional recovery of the grated vein.

A phospholipase A2 inhibitor (Ro 31-4493) prevents protein loss associated with the calcium paradox in isolated guinea pig hearts without effect on contracture, calcium overload, or the currents through L-type calcium channels.

OBJECTIVE: Although it is widely accepted that the hypercontraction provoked by the influx of Ca2+ on return to normal Tyrode solution after a period of Ca2+ depletion (the calcium paradox) contributes to the damage to the cell membrane, it remains possible that the Ca2+ overload activates intracellular enzymes that are important in the initial degradation of the cell membrane. This possibility was examined in this work. EXPERIMENTAL DESIGN: The effect of Ro 31-4493, a phospholipase A2 inhibitor, upon protein loss, hypercontracture, and ultrastructural changes in Langendorff perfused guinea pig hearts during the calcium paradox was studied. Because the degree of Ca2+ overload might also be affected, the action of the drug on the resting and action potentials and the whole cell currents through the L-type Ca2+ channels and the changes in [Ca2+]i in isolated guinea pig ventricular myocytes were also studied. RESULTS: Ro 31-4493, at 10-50 microM, inhibited the release of proteins from guinea pig Langendorff perfused hearts during the calcium paradox in a dose dependent way. This protective effect required preincubation with the agent before the Ca2+ depletion. No significant effect upon the contractile behaviour, as recognised from the sustained increase in intraventricular pressure on Ca2+ repletion, was produced. However, structural changes suggest that the extent of hypercontraction, the disruption of cell membrane and the release of mitochondria was less in treated hearts. Ro 31-4493 produced no change in the resting and action potentials or the behaviour of the L-type Ca2+ channels with either Ca2+ or Na+ as the charge carrier, while measurements of [Ca2+]i indicated a similar Ca2+ overload in both treated and untreated hearts. CONCLUSION: The effects of Ro 31-4493 are inconsistent with mechanical consequences of hypercontraction upon activation of a Ca2+ dependent phospholipases may be an important step.

Characteristics of L-aspartate transport and expression of EAAC-1 in sarcolemmal vesicles and isolated cells from rat heart.

OBJECTIVE: L-Aspartate is an important intermediary metabolite in the heart and has also been implicated in myocardial protection, but little is known about its transport across the cardiac sarcolemma. In this study we have tested the hypothesis that the high affinity sodium-dependent aspartate transporter, EAAC-1 is expressed in heart and have also characterised aspartate transport into the myocardium. METHODS: Characteristics of L-[14C]aspartate uptake into rat heart were investigated using sarcolemmal vesicles and isolated myocytes. The expression of EAAC-1 in the two preparations was also investigated by western blotting. RESULTS: The K(m) and V(max) of L-aspartate uptake was 9.78+/-0.7 microM and 1.17+/-0.27 pmol/mg/s in vesicles compared to 6.53+/-1.24 microM and 13.65+/-1.0 pmol/microl/s in cells. In vesicles, L-aspartate uptake was dependent on external sodium and internal potassium, and was rheogenic. In cells, L-aspartate uptake was also dependent on external sodium. Addition of unlabelled L- and D-aspartate and L-glutamate significantly inhibited L-[14C]aspartate uptake in both preparations but D-glutamate had no effect. An antibody to the aspartate transporter, EAAC-1 recognised a protein of appropriate size in both vesicles and cells. CONCLUSIONS: L-aspartate uptake in heart is mediated by a high affinity sodium-dependent transporter. This is accompanied by the expression in heart of EAAC-1. The physiological significance of this transporter with respect to aspartate utilisation in the heart is discussed.

Interdependence of intracellular taurine and sodium in guinea pig heart.

OBJECTIVE: The aim was to investigate the effects of raising intracellular taurine on the intracellular sodium activity (aNa1) in isolated guinea pig ventricular myocytes, and the effect of procedures that raise intracellular sodium on taurine concentration in the perfused guinea pig ventricular tissue. METHODS: Taurine was introduced into the sarcoplasm of isolated ventricular myocytes, either during cell isolation or by diffusion from a penetrating micropipette, and the effect on aNai was measured using an ion sensitive microelectrode. Guinea pig hearts, mounted on a Langendorff apparatus, were perfused with a variety of physiological media and the level of taurine in the ventricles determined using high pressure liquid chromatography. RESULTS: An increase in intracellular taurine caused by its presence during cell isolation or by diffusion from a micropipette significantly reduced the aNai of isolated myocytes at rest, during perfusion with Ca depleted solutions, or on inhibition of the Na pump. In the guinea pig ventricles, taurine at 13.0(SEM 0.6) mmol.kg-1 wet weight comprised up to 45% of the free amino acids; since plasma taurine was 64(13) mumol.litre-1, this means that in vivo a large outwardly directed gradient for taurine exists (equivalent to a free energy of 13.7 KJ.mol-1). Upon perfusion with Ca,Mg free Tyrode solution (which raises intracellular sodium markedly), a time dependent loss of taurine occurred. Both the rate of loss and the total amount lost were increased when the Na pump was also inhibited. This loss of tissue taurine was not due to release from dead or lysed cells, as it was antagonised by procedures known to reduce the rise of aNai during Ca depletion, was inhibited by beta alanine (an inhibitor of taurine transport), and the fall in tissue taurine was not correlated with the appearance of lactate dehydrogenase in the effluent. CONCLUSIONS: The data from isolated myocytes and perfused guinea pig hearts were consistent with the presence of a Na/taurine symport which is activated to cause efflux of Na and taurine when either rise above their physiological level.

Protection of hearts from reperfusion injury by propofol is associated with inhibition of the mitochondrial permeability transition.

OBJECTIVE: Diminishing oxidative stress may protect the heart against ischaemia-reperfusion injury by preventing opening of the mitochondrial permeability transition (MPT) pore. The general anaesthetic agent propofol, a free radical scavenger, has been investigated for its effect on the MPT and its cardioprotective action following global and cardioplegic ischaemic arrest. METHOD: Isolated perfused Wistar rat hearts were subjected to either warm global ischaemia (Langendorff) or cold St. Thomas' cardioplegia (working heart mode) in the presence or absence of propofol. MPT pore opening was determined using [3H]-2-deoxyglucose-6-phosphate ([3H]-DOG-6P) entrapment. The respiratory function of isolated mitochondria was also determined for evidence of oxidative stress. RESULTS: Propofol (2 micrograms/ml) significantly improved the functional recovery of Langendorff hearts on reperfusion (left ventricular developed pressure from 28.4 +/- 6.2 to 53.3 +/- 7.3 mmHg and left ventricular end diastolic pressure from 52.9 +/- 4.3 to 37.5 +/- 3.9 mmHg). Recovery was also improved in propofol (4 micrograms/ml) treated working hearts following cold cardioplegic arrest. External cardiac work on reperfusion improved from 0.42 +/- 0.05 to 0.60 +/- 0.03 J/s, representing 45-64% of baseline values, when compared to controls (P < 0.05). Propofol inhibited MPT pore opening during reperfusion, [3H]-DOG-6P entrapment being 16.7 vs. 22.5 ratio units in controls (P < 0.05). Mitochondria isolated from non-ischaemic, propofol-treated hearts exhibited increased respiratory chain activity and were less sensitive to calcium-induced MPT pore opening. CONCLUSION: Propofol confers significant protection against global normothermic ischaemia and during cold cardioplegic arrest. This effect is associated with less opening of mitochondrial MPT pores, probably as a result of diminished oxidative stress. Propofol may be a useful adjunct to cardioplegic solutions in heart surgery.

Inflammation, oxidative stress and postoperative atrial fibrillation in cardiac surgery.

Postoperative atrial fibrillation (POAF) is a common complication of cardiac surgery that occurs in up to 60% of patients. POAF is associated with increased risk of cardiovascular mortality, stroke and other arrhythmias that can impact on early and long term clinical outcomes and health economics. Many factors such as disease-induced cardiac remodelling, operative trauma, changes in atrial pressure and chemical stimulation and reflex sympathetic/parasympathetic activation have been implicated in the development of POAF. There is mounting evidence to support a major role for inflammation and oxidative stress in the pathogenesis of POAF. Both are consequences of using cardiopulmonary bypass and reperfusion following ischaemic cardioplegic arrest. Subsequently, several anti-inflammatory and antioxidant drugs have been tested in an attempt to reduce the incidence of POAF. However, prevention remains suboptimal and thus far none of the tested drugs has provided sufficient efficacy to be widely introduced in clinical practice. A better understanding of the cellular and molecular mechanisms responsible for the onset and persistence of POAF is needed to develop more effective prediction and interventions.

NADH fluorescence in isolated guinea-pig and rat cardiomyocytes exposed to low or high stimulation rates and effect of metabolic inhibition with cyanide.

In this study we investigated whether NADH fluorescence levels changed in response to low or high rates of electrical stimulation in single ventricular myocytes isolated from rat and guinea-pig hearts, either during a single contraction or upon sustained electrical stimulation of cells. NADH levels were determined from cell autofluorescence and cell length monitored using an edge-tracking device. NADH/NAD+ was obtained by addition of cyanide, 100% NADH, and carbonylcyanide-p-trifluoromethoxy phenylhydrazone (FCCP), 100% NAD+. Rat myocytes exhibited slightly higher resting fluorescence levels than guinea-pig cells; however, NADH/NAD+ was higher in rat than guinea-pig cells (P < 0.05), 24.3+/-4.3 (N = 17) vs 14.6+/-1.6 (N = 17), respectively. There was no change in NADH fluorescence during a single contraction when cells were stimulated at either low (0.2 Hz) or high (3 Hz) rates in either species. Furthermore, NADH levels did not change upon sustained stimulation at 3 Hz in either species. Metabolic blockade with cyanide induced a dose dependent rise in NADH fluorescence which was similar for both rat and guinea-pig myocytes and reached a maximum at > or = 1 mM of cyanide. Although a full recovery of NADH fluorescence was seen in both types of cells after brief exposure to cyanide, the rate of recovery was significantly slower in rat myocytes; times to 90% recovery were 110+/-29 sec, N = 6, and 264+/-50 sec, N = 6, for guinea-pig and rat cells, respectively. This work demonstrates that although rat and guinea-pig myocytes have different resting NADH/NAD+, their response to electrical stimulation is the same, whereas in response to metabolic inhibition subtle differences are seen.