Systematic Evaluation of Benchmark G4 Probes and G4 Clinical Drugs using three Biophysical Methods: A Guideline to Evaluate Rapidly G4-Binding Affinity.

G-quadruplex DNA structures (G4) are proven to interfere with most genetic and epigenetic processes. Small molecules binding these structures (G4 ligands) are invaluable tools to probe G4-biology and address G4-druggability in various diseases (cancer, viral infections). However, the large number of reported G4 ligands (>1000) could lead to confusion while selecting one for a given application. Herein we conducted a systematic affinity ranking of 11 popular G4 ligands vs 5 classical G4 sequences using FRET-melting, G4-FID assays and SPR. Interestingly SPR data globally align with the rankings obtained from the two semi-quantitative assays despite discrepancies due to limits and characteristics of each assay. In the whole, PhenDC3 emerges as the most potent binder irrespective of the G4 sequence. Immediately below PDS, PDC-360A, BRACO19, TMPyP4 and RHPS4 feature strong to medium binding again with poor G4 topology discrimination. More strikingly, the G4 drugs Quarfloxin, CX5461 and c-PDS exhibit weak affinity with all G4s studied. Finally, NMM and Cu-ttpy showed heterogeneous behaviors due, in part, to their physicochemical particularities poorly compatible with screening conditions. The remarkable properties of PhenDC3 led us to propose its use for benchmarking FRET-melting and G4-FID assays for rapid G4-affinity evaluation of newly developed ligands.

Surface plasmon resonance study of the interaction of N-methyl mesoporphyrin IX with G-quadruplex DNA.

Surface plasmon resonance (SPR) was used to investigate the interaction between N-methyl mesoporphyrin IX (NMM) and different G-quadruplex (G4) topologies. The study was associated with circular dichroism analysis (CD) to assess the topology of the G4s when they interacted with NMM. We demonstrate the high selectivity of NMM for the parallel G4 structure with a dissociation constant at least ten times lower than those of other G4 topologies. We also confirm the ability of NMM to shift the G4 conformation from both the hybrid and antiparallel topologies toward the parallel structure.

Influence of the SPR Experimental Conditions on the G-Quadruplex DNA Recognition by Porphyrin Derivatives.

Surface plasmon resonance (SPR) is a powerful technique to study the interactions of ligands with analytes and therefore a number of biosensor surfaces and injection methods have been developed so far. However, many experimental parameters can affect the interactions and consequently the affinity measurements. In particular, the interactions of positively charged analytes (often used for anionic nucleic acids targets) can be influenced by the sensing surfaces (e.g., negatively charged), leading to significant nonspecific interactions as well as regeneration problems. The aim of the present work is to investigate the effect of different parameters, including ionic strength, SPR biosensor (i.e., nature of the surfaces), and the injection method on the recognition of porphyrin G-quadruplex ligands. We demonstrate that the injection method does not influence the affinity whereas the ionic strength and the nature of the surface impact the recognition properties of the porphyrin for the G-quadruplex DNA. We also found that self-assembled monolayer coating surface presents many advantages in comparison with carboxymethylated dextran surface for SPR studies of G-quadruplex DNA/ligand interactions: (i) the electrostatic interaction with charged analytes is less important, (ii) its structure/composition is less sensitive to the ionic concentration and less prone to unspecific adsorption, (iii) it is easily homemade, and (iv) the cost is approximately 10 times cheaper.

[Sinus node dysfunction associated with cardiac sarcoidosis: A case report]. / Dysfonction sinusale au cours de la sarcoïdose cardiaque: à propos d'un cas.

INTRODUCTION: Sarcoidosis is a multisystemic granulomatous disease of unknown cause occurring in young adults. Cardiac sarcoidosis patients are at increased risk for atrioventricular blocks and ventricular arrhythmias. Sinus node dysfunction is scarcely reported. OBSERVATION: We report a case of cardiac sarcoidosis revealed by a sinus node dysfunction and focus on cardiac and thoracic imaging to guide diagnosis. CONCLUSION: Sinus node dysfunction may be the first manifestation of cardiac sarcoidosis. In unexplained sinus node dysfunction in young patients, advanced cardiac imaging is a key to cardiac sarcoidosis diagnostic. Early recognition of cardiac sarcoidosis enables to start immunosuppressive treatment and discuss implantable cardioverter defibrillator implantation.

Constrained G4 structures unveil topology specificity of known and new G4 binding proteins.

G-quadruplexes (G4) are non-canonical secondary structures consisting in stacked tetrads of hydrogen-bonded guanines bases. An essential feature of G4 is their intrinsic polymorphic nature, which is characterized by the equilibrium between several conformations (also called topologies) and the presence of different types of loops with variable lengths. In cells, G4 functions rely on protein or enzymatic factors that recognize and promote or resolve these structures. In order to characterize new G4-dependent mechanisms, extensive researches aimed at identifying new G4 binding proteins. Using G-rich single-stranded oligonucleotides that adopt non-controlled G4 conformations, a large number of G4-binding proteins have been identified in vitro, but their specificity towards G4 topology remained unknown. Constrained G4 structures are biomolecular objects based on the use of a rigid cyclic peptide scaffold as a template for directing the intramolecular assembly of the anchored oligonucleotides into a single and stabilized G4 topology. Here, using various constrained RNA or DNA G4 as baits in human cell extracts, we establish the topology preference of several well-known G4-interacting factors. Moreover, we identify new G4-interacting proteins such as the NELF complex involved in the RNA-Pol II pausing mechanism, and we show that it impacts the clastogenic effect of the G4-ligand pyridostatin.

[Fighting against unexplained sudden death]. / Mort subite, en quête d'une explication.

Sudden cardiac death, mostly related to ventricular arrhythmia, is a major public health issue, with still very poor survival at hospital discharge. Although coronary artery disease remains the leading cause, other etiologies should be systematically investigated. Exhaustive and standardized exploration is required to eventually offer specific therapeutics and management to the patient as well as his/her family members in case of inherited cardiac disease. Identification and establishing direct causality of the detected cardiac anomaly may remain challenging, underlying the need for a multidisciplinary and experimented team.

Administration of dietary prebiotics improves growth performance and reduces pathogen colonization in broiler chickens.

Dietary prebiotics are thought to be potentially important alternatives to antibiotic growth promoters in poultry production because of their beneficial performance and health effects. The administration of dietary prebiotics has been demonstrated to improve animal health, growth performance, and microbial food safety in poultry production. In this study, we evaluated the effects of Saccharomyces-derived prebiotic refined functional carbohydrates (RFC) with yeast culture on growth performance and gastrointestinal and environmental microbiota when administered in-feed and through drinking water to broiler chickens. Broilers were administered 2 doses of prebiotic in-feed through 42 d of production and prebiotic-treated water in the final 72 h. Administration of prebiotic RFC improved ADG and decreased cecal Campylobacter counts, while the high dose also increased final BW. Additionally, significant main effects of prebiotic RFC dose were observed with the high dose improving ADG and ADFI over the finisher phase and final BW. Although the effects were not significant, the prevalence of Campylobacter in the cecum after feed withdrawal was 17% lower when broilers were administered the high prebiotic dose, and recovery of Campylobacter from litter was up to 50% lower when broilers were administered prebiotic RFC. Our results suggest that co-administration of RFC with yeast culture as a prebiotic can be used to improve growth performance and reduce human foodborne pathogens in poultry.

[Management of patients with pacemakers or implantable cardioverter defibrillator undergoing radiotherapy]. / Prise en charge des patients porteurs de stimulateurs cardiaques ou défibrillateurs automatiques implantables traités par irradiation.

An increasing number of patients with cardiac implantable electronic devices benefit from radiotherapy, warranting specific collaborative management between both radiation oncologists and cardiologists. Interactions between electromagnetic fields, secondary particles and cardiac implantable electronic devices may result in transient and reversible malfunctions with significant consequences depending on the underlying cardiac pathology and the level of patient's cardiac implantable electronic devices dependency. Numerous international guidelines on patients' management have been proposed and all agree on a total cumulated dose limit at the battery of 5Gy and on the need for an initial as well as repeated evaluation over time, up to 6months after the last radiation. The analysis of the published data revealed relatively rare incidence of significant adverse events. The most recent international guidelines underline the feasibility and safety of radiotherapy for cardiac implantable electronic devices holders, with the need for systematic local protocol in all radiotherapy centers.

[Survey into cardiac pacing in the Ile-de-France area for the year 2001: evaluation of indications]. / Enquête sur la stimulation cardiaque en Ile-de-France pour l'année 2001: evaluation des indications.

UNLABELLED: A survey into the implantation of cardiac pacemakers during 2001 in the Ile-de-France area was carried out by the French National Insurance Medical Service in order to evaluate performance in all centres performing more than 10 primary implantations per year. METHODS AND RESULTS: In 2001, 12 centres out of a total of 49 performed less than 50 primary implantations per year, representing 5% of the total regional activity, which was estimated to be 6414 procedures. The remaining 95% of procedures were spread evenly among 8 high-throughput centres (> 200 primary implantations per year) and 29 medium-throughput centres (50 to 200 primary implantations per year). Indications for pacing were analysed retrospectively by a team of regulatory doctors on a sample of 2176 patients with reference to the ACC/AHA/NASPE guidelines. After examination of the medical records, the indication was ranked as being class I, II or Ill (absence of indication). A valid indication was lacking in 8.2% of cases. Sinus node dysfunction represented 74.6% of the non-indications, and this classification had the predictive factors of asymptomatic dysfunction, and treatment with anti-arrhythmic or bradycardic medication. The proportion of class III interventions was significantly lower in the high-throughput centres (5.8 vs 9.9%, p < 0.05). CONCLUSION: 8.2% of primary pacing procedures were not indicated and resulted principally from asymptomatic sinus node dysfunction.

Prognostic value of radionuclide angiography in patients with right ventricular arrhythmias.

BACKGROUND: The prognosis of patients with right ventricular (RV) arrhythmias remains uncertain. This study prospectively evaluated the prognostic value of RV and left ventricular (LV) involvement assessed by radionuclide angiography (RNA) as predictors for sudden death. METHODS AND RESULTS: Patients (n=188) with severe arrhythmias originating from the RV were followed up for a mean of 45+/-34 months. Data on clinical presentation, resting and stress ECG, signal-averaged ECG, 24-hour Holter monitoring, and programmed stimulation were collected along with RNA. Patients were classified as group I (n=82) with normal RNA or group II (n=106) with an abnormal RV suggestive of arrhythmogenic RV cardiomyopathy, classified as diffuse or localized disease, with or without associated LV abnormalities. During follow-up, 14 patients died suddenly, all in group II. None of the clinical and electrical data were predictive of death. An abnormal RNA study was a highly predictive factor for death (P<0.005), as well as the presence of LV abnormalities (P<0.01). CONCLUSIONS: The present study confirms that arrhythmogenic RV cardiomyopathy is a severe disease with a high risk for cardiac death. Evidence of RV abnormalities in patients presenting with RV arrhythmias is highly predictive for sudden death, as is its association with LV involvement.

Electrophysiological end point for catheter ablation of atrial fibrillation initiated from multiple pulmonary venous foci.

BACKGROUND: The end point for catheter ablation of pulmonary vein (PV) foci initiating atrial fibrillation (AF) has not been determined. METHODS AND RESULTS: Ninety patients underwent mapping during spontaneous or induced ectopy and/or AF initiation. Ostial PV ablation was performed by use of angiograms to precisely define targeted sites. Success defined by elimination of AF without drugs was correlated with the procedural end point of the abolition of distal PV potentials. A total of 197 arrhythmogenic PV foci (97%)-single in 31% and multiple in 69%-and 6 atrial foci were identified. A discrete radiofrequency (RF) application eliminated the PV potentials in 9 PV foci, whereas 2 foci from the same PV required RF applications at separate sites in 19 cases. In others, a wider region was targeted with progressive elimination of ectopy. In 49 patients, multiple sessions were necessary owing to recurrent or new ectopy. The clinical success rates were 93%, 73%, and 55% in patients with 1, 2, and > or =3 arrhythmogenic PV foci. Recovery of local PV potential and the inability to abolish it were significantly associated with AF recurrences (90% success rate with versus 55% without PV potential abolition). PV stenosis was noted acutely in 5 of 6 cases, remained unchanged at restudy, and was associated with RF power >45 W. CONCLUSIONS: Multiple PV foci are involved in initiation of AF, and elimination of PV muscle conduction is associated with clinical success.

Effect of inorganic versus organic selenium on hen production and egg selenium concentration.

A 28-d experiment using 288 Hy-Line W-36 laying hens was conducted to compare sodium selenite (SS) with Se-enriched yeast (SY). The Se from SS or SY was supplemented into a corn-soybean meal basal diet at 0, 0.15, 0.30, 0.60, or 3.00 ppm, and the basal diet was formulated to provide 0.82% lysine and 2,950 kcal/kg of ME. Each treatment was replicated 4 times with 2 cages of 4 hens per cage in each replicate. Hen production was assessed daily, and 2 eggs per replicate were collected every 4 d for whole-egg Se analysis. Albumen quality was assessed at 2 egg storage temperatures (7.2 vs. 22.2 degrees C) with the eggs collected on d 24 and 28, respectively. The percentage of dirty and cracked eggs was greater (P < 0.04) in hens fed SY than in those fed SS. Percentage hen-day production was not affected (P > 0.05) by diet. Albumen quality of eggs stored at 22.2 degrees C was improved (P < 0.04) in eggs from hens fed SS, but there was no difference (P > 0.05) in albumen quality of eggs stored at 7.2 degrees C. Egg weight was linearly increased (P < 0.01) by SY. Whole-egg Se levels were linearly increased (P < 0.01) as dietary Se level increased for both sources of Se, but eggs from hens fed SY had higher (P < 0.01) Se concentrations than those fed SS. The results from this experiment indicate that percentage hen-day production is not affected by Se source, and that SY increases egg Se concentrations more than SS.

A comparison of two sources of phytase in liquid and dry forms in broilers.

Research with corn-soybean meal diets was conducted to compare phytase sources in commercial broilers. A Ca to nonphytate P (nPP) ratio of 2.5:1 was maintained in all diets. Experiments 1 and 2 were conducted from d 4 to 13 (experiment 1) or d 9 to 23 post-hatching (experiment 2) in batteries. The 10 treatments used in both experiment were: Diets 1 to 4 = 0.20, 0.25, 0.30, or 0.35% nPP; Diets 5 to 7 = diet 1 plus 100, 200, or 300 phytase units/kg of diet from Natuphos (NAT); and Diets 8 to 10 = diet 1 plus 100, 200, or 300 phytase units/ kg of diet from Ronozyme (RON). As nPP levels increased, daily gain (ADG), feed intake (ADFI), gain:feed, and toe and tibia ash percentage were linearly increased (P < 0.06) in experiments 1 and 2. Incremental addition of phytase, regardless of source, linearly increased (P < 0.07) ADG and ADFI in experiment 1. Broilers fed NAT had higher (P < 0.07) toe ash percentage in experiment 1 and 2, and higher (P < 0.02) ADG and ADFI in experiment 2 than those fed RON. In experiment 3, 3,360 broilers were used to test 2 dry phytase products from 0 to 41 d posthatching in a 3-phase feeding program. The treatments were: Diet 1) adequate Ca and nPP; Diet 2) nPP reduced by 0.20%; Diets 3 to 5 = diet 2 plus 300, 500, or 750 phytase units/kg of diet from NAT; and Diets 6 to 8 = Diet 2 plus 300, 500, or 750 phytase units/kg of diet from RON. Broilers fed the adequate Ca and nPP diet had improved ADG and ADFI overall, and tibia ash weight and percentage in all growth phases (P < 0.03) compared with those fed the reduced Ca and nPP diets. Overall, ADG and ADFI were linearly increased (P < 0.05) by incremental phytase addition in experiment 3. Both NAT and RON produce similar growth and bone ash traits in commercial broilers.

[New energy sources for ablative methods]. / Nouvelles sources d'énergie dans les méthodes ablatives.

Radiofrequency current is the reference energy source for endocavitary ablation of arrhythmias. It is particularly well adapted for the ablation of focal arrhythmogenic substrates such as accessory pathways or foyers of automatism. Technological advances have made the lesions larger but the extension of the indications of percutaneous ablation to more complex substrates such as atrial fibrillation have justified the evaluation of alternative energies. The production of linear transmural lesions or deeper lesions which respect the parietal myocardial architecture and endocardial structure are a challenge for these energies. The capacity of functional mapping specific to cryogenics has provided this energy source with a clinical application for ablation of high risk structures whereas other energies, despite the chronicity of their experimental evaluation, are still at the stage of preliminary clinical trials with the sophisticated catheters in special indications.

[Long-term evaluation of endocavitary cryoablation of nodal reentry]. / Evaluation à long terme de la cryoablation endocavitaire dans le traitement des réentrées nodales.

Radiofrequency ablation is the reference treatment of refractory nodal reentry. Cryoablation has the advantage of having more modulable effects and minimises the risk of permanent atrioventricular block (AVB). Its immediate efficacy seems comparable to that of radiofrequency ablation but the long-term results are not well known. Endocavitary cryoablation of the slow pathway was undertaken in 26 patients (18 women) with an average age of 47.7 +/- 72.8 years with re-entrant nodal tachycardia refractory to medical therapy. The primary success rate was 92% (24 out of 26). On average, 2.6 +/- 2.2 (1 to 10) cryoablations at - 70 degrees C were delivered and were preceded by 6.4 +/- 4.5 (1 to 16) cryomappings to locate the site of the slow pathway. During cryomapping, 8 episodes of AVB were observed in 6 patients (6 second or third degree), all of which were revertible on rewarming. No cases of permanent AVB were observed. An oesophageal stimulation test of inducibility was performed on the 4th day in 21 patients, 16 of which were not reinducible. During follow-up of 355 +/- 194 days, 22 of the 26 patients (85%) had no recurrence of the arrhythmia. Two of the 24 primary successes had a recurrence, in addition to the two primary failures. Two of the four recurrences occurred in a non-sustained form which was less disabilitating for the patient and the recurrences were controlled in the 4 patients by antiarrhythmic therapy. These results suggest that cryoablation may be a reliable and effective long-term treatment of re-entrant nodal tachycardias. If confirmed in larger series in terms of efficacy and safety, cryoablation could become the treatment of choice of re-entrant nodal tachycardia.

Cellular electrophysiological effects of flecainide on human atrial fibres.

STUDY OBJECTIVE: The aim of the study was to examine the electrophysiological characteristics of human atrial specimens collected during heart surgery and to investigate the effects of the class I antiarrhythmic agent flecainide on their electrical activity. DESIGN: Atrial specimens were studied using standard microelectrode techniques, with and without superfused flecainide (5 x 10(-7) M) or the transient outward current inhibitor 4-aminopyridine (0.5 mM). EXPERIMENTAL MATERIAL: Atrial fragments 0.5-1.0 cm2 were obtained at operation from 34 patients, mean age 30 years. There was no history of previous atrial arrhythmia in any patient and drug therapy was stopped 24 h before surgery. MEASUREMENTS AND MAIN RESULTS: Two types of transmembrane action potential were identified: (1) triangular shaped potentials (group A, classically found in animal models); (2) potentials with a large plateau preceded by a notch (group B). The effect of flecainide was compared on the the two types of action potential. In both, flecainide lessened the depolarisation rate. In group B, but not in group A, it increased the action potential duration at 50% and 90% repolarisation (APD50, APD90) and the effective refractory period. The notch in group B action potentials is generated by transient outward currents (Ito). Inhibition of these currents, either by increasing the pacing rate or by adding 4-aminopyridine, limited the increase in APD50, APD90, and effective refractory period generated by the presence of flecainide. CONCLUSIONS: The effects of flecainide on the atrial repolarisation process depend on the shape of the action potential. These effects are more marked in cells with a plateau, where Ito is activated.

Intracellular and extracellular recordings of sinus node activity: comparison with estimated sinoatrial conduction times during pacemaker shifts in rabbit heart.

Sinoatrial conduction times, estimated by premature atrial stimulation, were compared with direct measurement of the sinoatrial conduction time in 15 isolated rabbit sinus node preparations before and after intrasinusal pacemaker shifts induced by cooling. Transmembrane potentials and surface electrograms were recorded from the sinus node and crista terminalis. Extracellular sinus node activity was recorded in five preparations. Mapping was performed at 38 degrees C and 35 degrees C to determine the site of the dominant pacemaker. The sinus cycle was significantly longer at 35 degrees C (319.4 ms vs 258.1 ms). Intracellular measured conduction time was significantly shorter (63.8 ms vs 70.4 ms) because of caudal shift of the dominant pacemaker. Estimated sinoatrial conduction time was significantly longer (110.3 ms vs 85.4 ms) owing to the depression of automaticity by the extrastimulus. Extracellular measured conduction time did not differ significantly from intracellular measured conduction time. These results suggest that intrasinusal pacemaker shift may explain inaccuracies in indirect estimations of sinoatrial conduction time by atrial pacing techniques. Extracellular recordings appear to be a better method of evaluating sinoatrial conduction times.

Evidence of mitochondrial impairment during cardiac allograft rejection.

NADH laser fluorimetry and mitochondrial oxigraphy were used to study myocardial oxidative energy metabolism during cardiac allograft rejection. Heterotopic cardiac transplantation was performed on Lewis rats; allografts (with Fischer rat donors) were compared with isografts (with Lewis rat donors). In vivo and in vitro assays were performed six days after transplantation. Myocardial NADH fluorescence was recorded in vivo from grafted hearts, at baseline; during brief, complete ischemia; and during reperfusion. Oxygen consumption of mitochondria isolated from both native and grafted hearts was determined. Neither baseline levels nor maximum ischemic levels of NADH fluorescence (F0 = k[NADH]) were found to be significantly different between allografts (0.45 +/- 0.05 to 0.87 +/- 0.10) and isografts (0.45 +/- 0.04 to 1.11 +/- 0.05). During recovery, the rate of fluorescence decrease was significantly lower in allografts than in isografts (0.024 +/- 0.001 vs. 0.038 +/- 0.002 delta F0.s-1, P less than 10(-3], indicating a lower rate of NADH reoxidation. In the presence of malate and glutamate substrates, mitochondrial O2 consumption was significantly lower in allografts than in isografts (30 +/- 9 vs. 100 +/- 15 nanoatoms O2. min-1.mg prot-1, P less than 10(-2]. These results indicate that mitochondrial oxidative metabolism was impaired during the rejection process. Such energy production disturbances may contribute to the dysfunction of rejecting hearts.

Efficacy and safety of septal and left-atrial linear ablation for atrial fibrillation.

Atrial fibrillation (AF), the most common of all sustained cardiac arrhythmias, is frequently resistant to antiarrhythmic drugs, and physicians have seen limited success with catheter ablation limited to the right atrium. As a result, the safety and efficacy of systematic biatrial linear ablation for drug resistant AF was investigated. Forty-four patients (54 +/- 7 years) underwent catheter ablation of daily drug-resistant AF. Two right-atrial lines (1 septal and 1 cavotricuspid) and 3-4 left-atrial lines were transseptally performed: 2 joining each superior pulmonary vein to the posterior mitral annulus and 1 interconnecting them. An additional left-atrial septal line from the right superior pulmonary vein (RSPV) to the foramen ovalis was performed in 23 patients. Radiofrequency was delivered with a conventional thermocouple-equipped ablation catheter or with an irrigated tip ablation catheter for resistant cases and for sparing the endocardium. Of the 44 patients, 25 (57%) were successfully treated without antiarrhythmic drugs. Twelve patients (27%) improved (<6 hours of AF per trimester under a previously ineffective drug) and 7 (16%) were considered treatment failures. Multiple sessions were required to ablate new left-atrial macro-reentry and initiating foci (2.7 +/- 1.3 procedures per patient). Five patients had a pericardial effusion and 1 each a pulmonary embolism, an inferior myocardial infarction, and a reversible cerebral ischemic event. One patient had thrombosis of the 2 left pulmonary veins. Despite a relatively high success rate, this procedure is too long, and the safely and efficacy need to be improved and applied to a broader range of patients.

Study of muscular and ventricular function in dynamic cardiomyoplasty: a ten-year follow-up.

BACKGROUND: The basic physiologic principle underlying cardiomyoplasty is long-term electrostimulation of a latissimus dorsi muscle (LDM) wrapped around the heart to obtain a phasic activity that could be integrated with ventricular kinetics. The aim of cardiomyoplasty is to prolong survival and to improve the quality of life of patients with severe chronic and irreversible myocardial failure by improving systolic contraction and correcting diastolic dysfunction. METHODS: To evaluate the long-term outcome of cardiomyoplasty, we investigated 82 patients electively undergoing this procedure in-our hospital. All patients had severe chronic heart failure that did not respond to optimal medical treatment. Patients had a mean age of 50 +/- 12 years (84% males). The cause of heart failure was ischemic (55%), idiopathic cardiomyopathy (34%), ventricular tumor (6%), and other (5%). The mean follow-up was 4.3 years. RESULTS: The mean New York Heart Association functional class improved after operation from 3.2 to 1.8. Average radioisotopic left ventricular ejection fraction increased from 17% +/- 6% to 28% +/- 3% (p < 0.05). Stroke volume index increased from 35 +/- 9 to 46 +/- 8 ml/beat/m2 (p < 0.05). The heart size remained stable at long term (evaluated by echo and computed tomography scanning). After cardiomyoplasty the number of successive hospitalizations resulting from congestive heart failure was reduced to 0.4 hospitalizations/patient/year (before operation 2.5, p < 0.05). Computed tomography scans showed at long-term a preserved LDM structure in 82% of patients who underwent operation. Survival probability at 7 years was 54% for the totality of patients, and 66% for patients who underwent operation in New York Heart Association functional class 3. Five patients underwent heart transplantation after cardiomyoplasty (mean delay 29 months), principally as a result of the natural evolution of their underlying heart disease, without major technical difficulties. CONCLUSIONS: Our 10-year clinical experience demonstrates that cardiomyoplasty increases ejection fraction, improves functional class, and ameliorates quality of life. Ventricular volumes and diameters remain stable long term. LDM structure is maintained long term if electrostimulation is performed avoiding excessive myostimulation. Patient selection is the most important determinant for early and late outcome. Late death in patients undergoing cardiomyoplasty is principally due to sudden death. Our future aim is to incorporate a cardioverter-defibrillator in the cardiomyostimulator, thus improving long-term results. Cardiomyoplasty may delay or prevent end-stage heart failure and the need for heart transplantation.