Fusogenic membrane glycoproteins induce syncytia formation and death in vitro and in vivo: a potential therapy agent for lung cancer.

Fusogenic membrane glycoproteins (FMGs) are viral envelope proteins, which bind surface receptors and induce fusion of the cell membrane. An FMG-transfected cell will fuse with neighbor cells, thus forming syncytia that die within 5 days. In this report, plasmids encoding for FMGs from Human Endogenous Retrovirus-W (HERV-W) was compared with Gibbon Ape Leukemia Virus (GALV) and feline endogenous virus RD-114 (RD). These plasmids were transfected in human non-small-cell lung cancer (NSCLC) cells in vitro or directly injected into tumors in mice. All FMGs induced the formation of syncytia containing around 50 cells. HERV-W or GALV FMGs decreased up to 80% of cell viability in vitro and inhibited tumor growth in vivo (60-70% reduction). In contrast, RD FMG was not efficient. Apoptosis played a role in the death of the syncytia, but addition of the caspase inhibitor Z-VAD-fmk had no effect, suggesting that apoptosis is not the only mechanism responsible for FMG-induced cell death. Altogether, our results demonstrate that even at very low transfection efficiency, the antitumor activity of HERV-W FMG is as effective as that of GALV in vitro and in vivo for the treatment of human lung tumors.

18F-FDG PET studies in patients with extratemporal and temporal epilepsy: evaluation of an observer-independent analysis.

UNLABELLED: The aim of this study was to evaluate an observer-independent analysis of 18F-fluorodeoxyglucose (FDG) PET studies in patients with temporal or extratemporal epilepsy. METHODS: Twenty-seven patients with temporal epilepsy and 22 patients with extratemporal epilepsy were included in the study. All patients with temporal epilepsy and 7 patients with extratemporal epilepsy underwent surgical treatment. In patients who showed significant postoperative improvement (temporal, n = 23; extratemporal, n = 6), the epileptogenic focus was assumed to be located in the area of surgical resection. In extratemporal epilepsy patients who did not undergo surgery, the focus localization was determined using a combination of semiology, ictal and interictal electroencephalography, [99mTc]ethyl cysteinate dimer SPECT, MRI and [11C]flumazenil PET. Visual analysis was performed by two experienced and two less experienced blinded observers using sagittal, axial and coronal images. In the automated analysis after anatomic standardization and generation of three-dimensional stereotactic surface projections (SSPs), a pixelwise comparison of 18F-FDG uptake with an age-matched reference database (n = 20) was performed, resulting in z score images. Pixels with the maximum deviation were detected, summarized and attached to one of 20 predefined surface regions of interest. For comparison with 18F-FDG PET and MR images, three-dimensional overlay images were generated. RESULTS: In patients with temporal epilepsy, the sensitivity was comparable for visual and observer-independent analysis (three-dimensional SSP 86%, experienced observers 86%-90%, less experienced observers 77%-86%). In patients with extratemporal epilepsy, three-dimensional SSP showed a significantly higher sensitivity in detecting the epileptogenic focus (67%) than did visual analysis (experienced 33%-38%, each less experienced 19%). In temporal lobe epilepsy, there was moderate to good agreement between the localization found with three-dimensional SSP and the different observers. In patients with extratemporal epilepsy, there was a high interobserver variability and only a weak agreement between the localization found with three-dimensional SSP and the different observers. Although three-dimensional SSP detected multiple lesions more often than visual analysis, the determination of the highest deviation from the reference database allowed the identification of the epileptogenic focus with a higher accuracy than subjective criteria, especially in extratemporal epilepsy. CONCLUSION: Three-dimensional SSP increases sensitivity and reduces observer variability of the analysis of 18F-FDG PET images in patients with extratemporal epilepsy and is, therefore, a useful tool in the evaluation of this patient group. The benefit of this analytical approach in patients with temporal epilepsy is less apparent.

Delayed rectifier potassium current in undiseased human ventricular myocytes.

OBJECTIVE: The purpose of the study was to investigate the properties of the delayed rectifier potassium current (IK) in myocytes isolated from undiseased human left ventricles. METHODS: The whole-cell configuration of the patch-clamp technique was applied in 28 left ventricular myocytes from 13 hearts at 35 degrees C. RESULTS: An E-4031 sensitive tail current identified the rapid component of IK (IKr) in the myocytes, but there was no evidence for an E-4031 insensitive slow component of IK (IKs). When nifedipine (5 microM) was used to block the inward calcium current (ICa), IKr activation was fast (tau = 31.0 +/- 7.4 ms, at +30 mV, n = 5) and deactivation kinetics were biexponential and relatively slow (tau 1 = 600.0 +/- 53.9 ms and tau 2 = 6792.2 +/- 875.7 ms, at -40 mV, n = 7). Application of CdCl2 (250 microM) to block ICa altered the voltage dependence of the IKr considerably, slowing its activation (tau = 657.1 +/- 109.1 ms, at +30 mV, n = 5) and accelerating its deactivation (tau = 104.0 +/- 18.5 ms, at -40 mV, n = 8). CONCLUSIONS: In undiseased human ventricle at 35 degrees C IKr exists having fast activation and slow deactivation kinetics; however, there was no evidence found for an expressed IKs. IKr probably plays an important role in the frequency dependent modulation of repolarization in undiseased human ventricle, and is a target for many Class III antiarrhythmic drugs.

Determinants of early and late results of combined valve operations and coronary artery bypass grafting.

BACKGROUND: Factors determining the outcome of operative correction of valvular abnormalities combined with coronary artery bypass grafting are still incompletely defined. METHODS: Determinants of early and late (more than 90 days) deaths and event-free survival were studied for combined valve operations and coronary artery bypass grafting in 741 patients using multivariate analysis. RESULTS: Ninety-day survival probability was 89% (95% confidence interval, 87% to 92%). Preoperative risk factors for early death were age, female sex, renal failure, New York Heart Association class IV or V, and mitral insufficiency. The operative risk factor was the duration of aortic cross-clamping. Five- and 10-year survival probabilities were 74% (95% confidence interval, 71% to 78%) and 43% (95% confidence interval, 36% to 50%), respectively. Preoperative risk factors for late death were age, preoperative renal failure, New York Heart Association class IV or V, vessel disease, and nonsinus rhythm. Five- and 10-year event-free survival probabilities were 57% (95% confidence interval, 53% to 61%) and 23% (95% confidence interval, 17% to 28%), respectively. Preoperative risk factors for non-event-free survival were age, female sex, reduced left ventricular function, mitral regurgitation, and pacemaker rhythm. CONCLUSION: The demographic factors of age and female sex; the comorbid condition of renal failure; the cardiac conditions of advanced New York Heart Association class, left ventricular function, mitral regurgitation, vessel disease, and cardiac rhythm; and the operative condition of ischemia time are the most important predictors of clinical outcome after combined valve operations and coronary artery bypass grafting.

Mitral valve surgery combined with coronary bypass grafting: multivariate analysis of factors predicting early and late results.

Total of 123 patients (mean age: 63.8 +/- 7.3 years, (male/female 65/35 percent) underwent mitral valve surgery combined with coronary artery bypass grafting during a seven year period. Preoperatively 12% of them belonged to NYHA functional class II, 54% to class III, 29% to class IV and 3% was operated under emergency conditions. The mitral valve lesion was most frequently either ischaemic (45%) or rheumatic (33%) in origin. Left ventricle function was moderately decreased in 18% and severely damaged in 3% of the patients as documented by preoperative ventriculography. Coronary surgery was performed in all cases with an average number of distal anastomosis of 2.2 +/- 1.1 per patient. The hospital mortality was 13%. Risk factors for early and late mortality were determined by univariate and multivariate analysis. Advanced preoperative functional class and decreased left ventricular function or ischaemic etiology were identified as significant risk factors for early mortality. The patients were followed for an average of 33 +/- 25 months. The majority of them experienced significant functional improvement postoperatively with 69% belonging to NYHA class I or II. The late survival for the 107 hospital survivors was 94.7% at one year, and 84.7% at five years, respectively. Late survival was independently determined by preoperative functional class or previous myocardial infarction. Freedom from ischemic and valve related complications at five years was 95% and 71.2% respectively. 58.2% of the hospital survivors were in functional class I or II and free of any valve related or ischemic complications at the end of the fifth follow up year.(ABSTRACT TRUNCATED AT 250 WORDS)

Aortic and mitral valve replacement with the Carpentier-Edwards pericardial bioprosthesis: mid-term clinical results.

Between May 1st 1985 and December 31st 1992, 283 Carpentier-Edwards pericardial valves were implanted in 260 patients in the aortic (n = 196), mitral (n = 41) and both aortic and mitral (n = 23) positions at the Gasthuisberg University Hospital in Leuven, Belgium. Patients undergoing tricuspid valve replacement or mixed replacement with another type of prosthesis were excluded from this study. The mean age was 70 +/- 7 years, there were 121 males and 139 females. The mean follow up was 40.55 months, the total follow up experience 10543 months (878.6 patients years). Hospital mortality was 10.4%, and was not significantly related to the position of the valve: 17.3% +/- 7.88% (n = 23) for double valve replacement, 10.2% +/- 2.16% (n = 196) for aortic valve replacement and 7.3% +/- 4.06% (n = 41) for mitral valve replacement. Hospital mortality was 14.1% +/- 3.27% for those with and 7.48% +/- 2.5% for those without concomitant coronary surgery (p = NS). Survival at 92 months was 63% +/- 6% and was not significantly related to the position of the valve. Not a single patient needed to be reoperated because of primary tissue failure. We conclude that the mid-term durability of this valve is excellent and consider the Carpentier-Edwards pericardial valve as the stented bioprosthesis of choice both in the aortic and mitral positions for the elderly. Because of the older age of our study population and the medium term length of follow up, we were unable to draw any conclusions concerning the incidence of calcific degeneration of this valve.

Combined valve and coronary artery bypass surgery: early and late results.

Determinants of early, late and event-free survival of combined valve and coronary artery bypass graft (CABG) surgery were studied in 420 patients using multivariate analysis. It was found that the risk of hospital death increases 5 times when the preoperative NYHA class was > or = IV, 3 times when left ventricular (LV) function is significantly impaired and is double when mitral regurgitation is present. The survival probability of hospital survivors was 91% (87.3-94.5%) at 5 years. Late mortality was determined by advanced preoperative NYHA class ( > or = IV) and the presence of mitral regurgitation. The event-free survival probability of hospital survivors, i.e. total events including death, valve-related complications, ischemic complications and recurrent NYHA class > or = IV, was 73.0% (66.7-79.5%) at 5 years. Postoperative events were determined by the presence of preoperative NYHA class > or = IV, impaired ventricular function, mitral regurgitation and non-sinus rhythm. It is concluded that these parameters can be considered as the most important predictors of clinical outcome after combined valve and CABG surgery.

Nucleoside transport inhibition mediates lidoflazine-induced cardioprotection during intermittent aortic crossclamping.

The effects of pretreatment with the nucleoside transport inhibitor lidoflazine on repeated ischemia-reperfusion injury induced by normothermic intermittent aortic crossclamping were studied in canine hearts. Eighteen mongrel dogs were allocated to three groups: placebo (n = 6), lidoflazine (1 mg/kg) (n = 6), and lidoflazine (1 mg/kg) plus the adenosine receptor blocker aminophylline (7 mg/kg) (n = 6). Pretreatment was performed intravenously during 15 minutes before extracorporeal circulation. All hearts were subjected to four intervals of 15 minutes of global ischemia each followed by 10 minutes of reperfusion. After weaning from extracorporeal circulation, functional recovery was followed for 1 hour. In the lidoflazine group, myocardial adenosine content (0.25 +/- 0.06 mumol/gm dry weight) was 3.5 times higher than that in the control group (0.07 +/- 0.03 mumol/gm dry weight; p < 0.05) at the end of the last aortic crossclamping. The release of adenosine from the myocardium during each reperfusion period was significantly higher than that in the control group (p < 0.05). Myocardial extraction of lactate was normalized at every reperfusion interval in the lidoflazine group but not in the control group (p < 0.05). In the lidoflazine group functional recovery was significantly better than that in the control group. Positive rate of rise of pressure, negative rate of rise of pressure, and cardiac output recovered to, respectively, 150% +/- 19%, 82% +/- 8%, and 131% +/- 15% in the lidoflazine group versus, respectively, 37% +/- 9%, 23% +/- 7%, and 29% +/- 8% in the control group (p < 0.001) at 1 hour after extracorporeal circulation. When the adenosine receptor blocker aminophylline was administered in association with lidoflazine, protection dropped significantly: positive and negative rate of rise of pressure and cardiac output were, respectively, 58% +/- 8%, 46% +/- 9%, and 67% +/- 16% at 1 hour after extracorporeal circulation (p < 0.05 versus lidoflazine alone). These results suggest that the cardioprotective effects of lidoflazine are at least in part mediated by adenosine receptor stimulation via nucleoside transport inhibition-induced accumulation of endogenous adenosine in the myocardium.

Time course of serum Lp(a) in men after coronary artery bypass grafting.

The serum Lp(a) time course was studied in 100 male patients who underwent coronary artery bypass grafting (CABG). The patients were randomized in a placebo (N = 50) and pravastatin treated (N = 50) group. The pravastatin regimen was 10 mg daily from the third postoperative day on and 20 mg daily after 1 week during 11 weeks. Lp(a) levels and serum lipids were analyzed at baseline, at 3 and 10 days, and at 4 and 12 weeks post-CABG. A decrease of serum Lp(a) levels at the third postoperative day was seen which parallels the changes noted with the other serum lipids when using extracorporeal circulation. In contrast with the other serum lipids, a slight but significant Lp(a) overshooting was noticed at day 10 followed by a decrease of the serum Lp(a) levels to preoperative levels 1 month after the acute event. The study clearly depicts that there is a significant time-dependent effect on the serum Lp(a) levels post-CABG and that there is no effect of treatment (pravastatin). The data also reveal that reliable postoperative Lp(a) measurements can be made at earliest 1 month post-CABG.

Time course of serum lipid and lipoprotein levels after coronary bypass surgery: modification by pravastatin.

The time course of the changes in serum lipid and lipoprotein levels have been studied in 100 patients undergoing coronary artery bypass surgery. Marked decreases occur in the levels of cholesterol (-45%), HDL-cholesterol (-35%), LDL-cholesterol (-53%), Apo A1 (-43%) and Apo B (-43%) on the third day after the operation. In the control group the values gradually returned to preoperative levels which are reached after three months. Three months after the intervention, in the pravastatin-treated group (20 mg/day) the values of total cholesterol (-20%), LDL-cholesterol (-28%) and Apo B (-22%), remained significantly lower than in the control group (p 0.0001) and were lower over the whole duration of the study. The postoperative time course of HDL-cholesterol and Apo A1, however, was not significantly influenced by the administration of pravastatin.

[Effect of acute aprotinin (Gordox) therapy on hemostasis in heart surgery patients, with special reference to hyperfibrinolysis]. / Az akut aprotinin (Gordox) terápia hatása a haemostasisra szívsebészeti betegekben, különös tekintettel a hyperfibrinolysisre.

Authors have studied the effect of Gordox-therapy on haemostasis after open heart surgery in a prospective clinical trial. Thirty seven patients (pts) undergoing cardiac surgery due to their valve disease were randomly assigned either to control-group (20 pts) or to Gordox-group (17 pts). The patients in the Gordox group were given Gordox according the following scheme: 2 M IU within 20 min. after induction of anaesthesia followed by 0.5 M IU/hour infusion until the end of the operation. One M IU also was given into the oxygenator before starting the extracorporeal circulation. The postoperative chest tube drainage was less in Gordox-group (534 +/- 260 ml vs. 987 +/- 583 ml, p less than 0.005), and donor blood and fresh frozen plasma requirement was also lower in this group (534 +/- 633 ml vs. 935 +/- 718 ml p less than 0.05; 70 +/- 153 ml vs. 211 +/- 245 ml p less than 0.05, respectively). There was no significant difference between the two groups concerning the postoperative activated partial thromboplastin time, prothrombin time, thrombin time values. The authors could document significantly higher fibrinogen concentration and significantly lower fibrinolytic activity postoperatively in the Gordox-group (p less than 0.05). Gordox therapy has advantageous effect on haemostasis after open heart surgery which can be documented both by clinical and laboratory examination.

A study of plasma and cardiac tissue concentrations of ceftriaxone (Rocephin) in open heart surgery.

Postoperative infection following cardiac surgery procedures is a serious complication. Cephalosporines are frequently used in prophylaxis. The aim of this study was to determine whether adequate therapeutic concentrations of ceftriaxone were present in cardiac tissues and plasma after 2 g single dose administered intravenously. Twenty-four consecutive patients undergoing open heart surgical procedures enrolled in the study. The antibiotic concentration was measured by agar well diffusion method. Samples of plasma were taken simultaneously of tissue removal in order to determine the antibiotic penetration. Tissue concentrations in the cardiac appendage, in the mitral and aortic valves and in the vessels of the graft exceeded MIC of the anticipated pathogens and especially high concentration was detected in pericardiac tissue during the time of operation. Therapeutic level was achieved against only certain strains of Pseudomonas.

Early clinical experience with a simple method for autotransfusion in cardiac surgery.

A simple method for intraoperative autotransfusion (ATF) in open-heart surgery was tested in a prospective clinical trial. The patients were randomly assigned to a control group (33) or to the ATF group (35). The intraoperative ATF was combined with preoperative collection of blood and postoperative ATF. The postoperative chest-tube drainage was reduced by 24.3%, the donor-blood requirement by 43.3% and the consumption of fresh-frozen plasma by 43.9% in the ATF group as compared with the controls (all differences statistically significant). To investigate possible haematologic side effects of ATF, measurements of haemoglobin, haematocrit, fibrinogen concentration, thrombin, prothrombin and partial thromboplastin time, antithrombin-III and fibrinolytic activity were made in all patients preoperatively and on postoperative days 1 and 2. No statistical differences were then found between the controls and the ATF group. Microbiologic tests of blood sampled from the cardiotomy reservoir gave satisfactory results.

Valve surgery combined with myocardial revascularisation. Report of 62 cases.

Combined valvular and coronary bypass surgery was performed in 1984-1988 in 62 patients. Their age range at operation was 38-75 (mean 59) years and more than 90% were in NYHA class III or IV. Valve replacement was performed in 55 cases--mitral in 13, aortic in 33, tricuspid in one and combined valve procedures in eight cases--and valvular correction in seven cases. The patients also received a total of 92 coronary artery grafts (1-4/patient, mostly as single grafts). The hospital mortality was 3% and there were two late deaths. Follow-up was 100% complete 2-52 (mean 22) months after surgery, and more than 90% of the survivors were improved.

A simple method for intraoperative autotransfusion in cardiac surgery.

Authors present a simple, safe and inexpensive method of intraoperative autotransfusion (ATF) for open-heart surgery. A brief comparison is given with other techniques already in use.

Fatty acid peptide derivatives as model compounds to protect elastin against degradation by elastases.

Peptide sequences which fit the extended binding sites of porcine pancreatic elastase and human leukocyte elastase were covalently coupled to oleic acid. These compounds behave as competitive inhibitors towards both elastases. The coupling of fatty acid moiety to the peptide greatly decreases its inhibitor constant (Ki) vs human leukocyte elastase (Ki for Oleoyl(Ala)2ProValine: 3.0 (10(-6)M). It is less active on porcine pancreatic elastase (Ki for Oleoyl(Ala)2ProAlanine: 3.8 10(-4)M). The modifications of the carboxylic end group of the peptide to an aldehyde further greatly enhanced the inhibition capacity of the compound towards leukocyte elastase (Ki for Oleoyl(Ala)2ProAlaninal: 0.7 microM). Oleoyl peptide derivatives were seen to bind in a saturable fashion to purified insoluble elastin, and decreased the susceptibility of the macromolecule to hydrolysis by both pancreatic and leukocyte elastases. As stoichiometric quantities of elastase (vs inhibitor) could not desorb 3H-oleoyl(Ala)2Pro-Val bound to insoluble elastin, it is postulated that oleoyl peptide derivatives may act as bifunctional agents. This contention was further strengthened by the comparison of the adsorption curves of elastase to untreated insoluble elastin and elastin saturated with oleoyl peptide derivatives respectively. It was shown finally that Oleoyl(Ala)2Pro-Valine was also capable of inhibiting elastases in their adsorbed form to insoluble elastin.