Search

Inosine, a break-down product of adenosine, has been recently shown to exert inodilatory and anti-inflammatory properties. We investigated the effects of inosine on ischemia/reperfusion injury in a rat heart transplantation model.Intraabdominal heterotopic transplantation was performed in Lewis rats. After 1h of ischemic preservation, reperfusion was started after application of either saline vehicle (control, n=12) or inosine (100 mg/kg, n=12). Coronary blood flow, left ventricular function, endothelium-dependent vasodilatation to acetylcholine and endothelium-independent vasodilatation to sodium nitroprusside, and high energy phosphate content were measured after 1 and 24h of reperfusion. In addition, the activation of the poly(ADP-ribose) polymerase was detected by immunhistology.After 1h, coronary blood flow (4.1+/-0.3 ml/(ming) vs 2.9+/-0.3 ml/(ming), p0.05), left ventricular systolic pressure (102+/-9 mmHg vs 83+/-4 mmHg, p0.05) and dP/dt (2765+/-609 mmHg/s vs 1740+/-116 mmHg/s, p0.05) were significantly higher in the inosine group in comparison to control. Vasodilatatory response to sodium nitroprusside was similar in both groups. Acetylcholine resulted in a significantly higher increase in coronary blood flow in the inosine group (76+/-5% vs 48+/-9%, p0.05). Energy charge potential was significantly higher in the inosine group (1.69+/-0.10 micromol/g vs 0.74+/-0.27 micromol/g, p0.05). After 24h, there was no difference between the groups in basal coronary blood flow, left ventricular systolic pressure, dP/dt, and the response to sodium nitroprusside. However, acetylcholine led to a still significantly higher response in the inosine group (112+/-13% vs 88+/-7%, p0.05). Immunhistologic stainings revealed activation of poly(ADP-ribose) polymerase in control animals which was abolished by inosine.Thus, inosine improves myocardial and endothelial function during early reperfusion after heart transplantation with a persisting beneficial effect against reperfusion induced graft coronary endothelial dysfunction. The effects of inosine are mediated at least partly by modulation of the peroxynitrite-poly(ADP-ribose) polymerase pathway. more...

Hadron therapy has already proven to be successful in cancer therapy, and might be a noninvasive alternative for the ablation of cardiac arrhythmias in humans. We present a pilot experiment investigating acute effects of a 12C irradiation on the AV nodes of porcine hearts in a Langendorff setup. This setup was adapted to the requirements of charged particle therapy. Treatment plans were computed on calibrated CTs of the hearts. Irradiation was applied in units of 5 and 10 Gy over a period of about 3 h until a total dose of up to 160 Gy was reached. Repeated application of the same irradiation field helped to mitigate motion artifacts in the resulting dose distribution. After irradiation, PET scans were performed to verify accurate dose application. Acute AV blocks were identified. No other acute effects were observed. Hearts were kept in sinus rhythm for up to 6 h in the Langendorff setup. We demonstrated that 12C ions can be used to select a small target in the heart and, thereby, influence the electrical conduction system. Second, our pilot study seems to suggest that no adverse effects have to be expected immediately during heavy ion irradiation in performing subsequent experiments with doses of 30-60 Gy and intact pigs. more...

The aim of the present study was to investigate the effects of the novel poly(ADP-ribose) polymerase (PARP) inhibitor PJ34 ( N -(6-oxo-5,6-dihydro-phenanthridin-2-yl)- N , N -dimethylacetamide) on myocardial and endothelial function after hypothermic ischemia and reperfusion in a heterotopic rat heart transplantation model. After a 1-hour ischemic preservation, reperfusion was started either after application of placebo or PJ34 (3 mg/kg). The assessment of left ventricular pressure–volume relations, total coronary blood flow, endothelial function, myocardial high energy phosphates, and histological analysis were performed at 1 and 24 hours of reperfusion. After 1 hour, myocardial contractility and relaxation, coronary blood flow, and endothelial function were significantly improved and myocardial high energy phosphate content was preserved in the PJ34-treated animals. Improved transplant function was also seen with treatment with another, structurally different PARP inhibitor, 5-aminoisoquinoline. The PARP inhibitors did not affect baseline cardiac function. Immunohistological staining confirmed that PJ34 prevented the activation of PARP in the transplanted hearts. The activation of P-selectin and ICAM-1 was significantly elevated in the vehicle-treated heart transplantation group. Thus, pharmacological PARP inhibition reduces reperfusion injury after heart transplantation due to prevention of energy depletion and downregulation of adhesion molecules and exerts a beneficial effect against reperfusion-induced graft coronary endothelial dysfunction. more...

Background— Particle therapy, with heavy ions such as carbon-12 ( 12 C), delivered to arrhythmogenic locations of the heart could be a promising new means for catheter-free ablation. As a first investigation, we tested the feasibility of in vivo atrioventricular node ablation, in Langendorff-perfused porcine hearts, using a scanned 12C beam. Methods and Results— Intact hearts were explanted from 4 (30–40 kg) pigs and were perfused in a Langendorff organ bath. Computed tomgraphic scans (1 mm voxel and slice spacing) were acquired and 12 C ion beam treatment planning (optimal accelerator energies, beam positions, and particle numbers) for atrioventricular node ablation was conducted. Orthogonal x-rays with matching of 4 implanted clips were used for positioning. Ten Gray treatment plans were repeatedly administered, using pencil beam scanning. After delivery, positron emission tomography-computed tomgraphic scans for detection of β + ( 11 C) activity were obtained. A 12 C beam with a full width at half maximum of 10 mm was delivered to the atrioventricular node. Delivery of 130 Gy caused disturbance of atrioventricular conduction with transition into complete heart block after 160 Gy. Positron emission computed tomgraphy demonstrated dose delivery into the intended area. Application did not induce arrhythmias. Macroscopic inspection did not reveal damage to myocardium. Immunostaining revealed strong γH2AX signals in the target region, whereas no γH2AX signals were detected in the unirradiated control heart. Conclusions— This is the first report of the application of a 12 C beam for ablation of cardiac tissue to treat arrhythmias. Catheter-free ablation using 12C beams is feasible and merits exploration in intact animal studies as an energy source for arrhythmia elimination. more...

Previous studies have demonstrated hemodynamic instability and cardiac dysfunction in the brain-dead organ donor. It remains unclear if primary cardiac dysfunction is responsible for hemodynamic deterioration or decreased cardiac function is secondary to brain death- associated altered loading conditions. Therefore in the present study the effects of brain death on hemodynamics and cardiac function were analyzed in vivo in an open chest model and ex vivo in a cross-circulated heart preparation. In a second protocol, the impact of brain death- associated hemodynamic changes on postischemic graft function was investigated. Brain death was induced injecting saline in a subdural Foley catheter. Induction of brain death led to a hyperdynamic reaction followed by hemodynamic deterioration with a decrease of systemic vascular resistance and myocardial contractility. If the hearts were explanted and assessed ex vivo, no differences were found between control and brain-dead hearts. Furthermore, both control and brain-dead hearts showed full functional recovery after 4 hours of hypothermic ischemic storage. Despite hemodynamic deterioration in situ after brain death, there were no differences between the postischemic function of control and brain-dead hearts. These results indicate that myocardial dysfunction is not irreversible and may be secondary to altered loading conditions, and that the recovery of cardiac function after long-term hypothermic storage is not impaired by the hemodynamic changes observed in situ after brain death induction. These data may also indicate that potential donor hearts might not be excluded from transplantation on the basis of impaired hemodynamic characteristics, especially if they are evaluated by load- dependent parameters. Perioperative donor heart performance is an important issue of cardiac transplantation. About 20% of potential donor hearts must be rejected on the basis of poor primary cardiac function or hemodynamic instability. On the other hand, the major cause of death after transplantation during the early postoperative period is graft failure, mostly in association with pulmonary hypertension (1). Recent studies suggest that not pulmonary hypertension per se but preexistent myocardial damage (2) as a result of brain death or ischemia-reperfusion injury in association with pulmonary hypertension may cause graft dysfunction (3, 4). A number of experimental and clinical studies report hemodynamic instability in the donor organism following brain death (3, 5-7), but it remains unclear which mechanisms lead to hemodynamic collapse in brain-dead organ donors. Catecholamine injury and hormone depletion are suggested to play a major role in cardiac dysfunction (3, 5). In contrast, some studies demonstrated that neurohumoral factors may have only minor importance in brain death-associated cardiac dysfunction (8, 9). Moreover, there is no evidence in the literature that the heart is the primary target of the detrimental effects of brain death resulting in hemodynamic instability or that myocardial dysfunction is secondary to changed loading condi- tions (6, 10, 11). It remains also unclear if changes of cardiac function are reversible or irreversible. A further issue of contro- versial discussion is the impact of brain death-associated changes on the ischemic tolerance of the donor hearts and posttransplant graft function. In the present study the effects of brain death on hemodynamics and cardiac function were analyzed in vivo in an open chest model and ex vivo in a cross-circulated heart preparation. The latter was used to exclude any load-dependent effect that may have an influence on cardiac function. In a second protocol, the impact of brain death-associated hemodynamic changes on postischemic graft function was investigated. more...

BACKGROUND We studied the effect of the calmodulin antagonist trifluoperazine (TFP) on isolated intact rat tracheal and pulmonary artery smooth muscle contractile behaviour. METHODS Experimental series: 1) TFP-dose-response curves for TFP's effect on force generation were constructed using rat tracheal smooth muscles and rat pulmonary artery preparations (n = 8). A concentration of 1 micromol/l TFP was chosen for the subsequent experimental series. 2) Tracheas and pulmonary arteries (n = 14) were dissected in three segments. One of them was used immediately for experiments ("native"), the other two were treated for 12 h in 4 degrees C Tyrode solution without ("12 h cold storage") or with 1 micromol/l TFP ("12 h cold storage + TFP"). These preparations contracted after supramaximal effective electrical field stimulation. The force-clamping technique was used to analyse kinetic and mechanical parameters of smooth-muscle contraction in both types of preparation (measurement conditions: resting tension 2 mN, 37 degrees C, modified Krebs-Henseleit solution). RESULTS 1) TFP decreased developed force dose-dependently in pulmonary artery and tracheal smooth muscle. 2) During sustained tonic activation, the contraction kinetics become slower both with and without TFP treatment (p < 0.0001). 3) TFP caused a dramatic retardation of the velocity of force generation in both types of preparation for any given time interval during the course of a tonic activation (p < 0.0005). 4) The dramatic effects of TFP on the contraction kinetics were not associated with effects on the extent of force generation. CONCLUSIONS These results support the assumption that tracheal and pulmonary artery smooth muscle cross-bridge rates are controlled by a calcium-calmodulin-dependent myosin light chain kinase. This finding suggests the involvement of a calmodulin-independent regulator process responsible for the changes observed in the cross-bridge cycling rates during sustained tonic activation. A direct intervention on the contractile apparatus level is a measure for reduction of smooth-muscle tone without negative inotropic side effects. more...