Effects of plasma membrane Ca2+-ATPase tyrosine phosphorylation on human platelet function

BACKGROUND: The plasma membrane Ca(2+)-ATPase (PMCA) plays an essential role in maintaining low intracellular Ca(2+) ([Ca(2+)](i)) in resting platelets. Earlier studies demonstrated that platelet activation by thrombin results in tyrosine phosphorylation of PMCA, which inhibits pump activity. OBJECTIVES: The objective was to determine the functional consequences of PMCA tyrosine phosphorylation. METHODS: A decapeptide including the tyrosine phosphorylation site of PMCA and a scrambled version were synthesized and introduced into human platelets using saponin. Fura-2 calcium monitoring and aggregometry were used to characterize the effects of inhibition of tyrosine phosphorylation. RESULTS: Western blot analysis of immunoprecipitates showed that introduction of the inhibitory peptide decreased tyrosine phosphorylation of PMCA by nearly 60% in saponin-permeabilized, thrombin-treated platelets as compared with the scrambled control peptide. Concomitant with inhibition of PMCA tyrosine phosphorylation was a significant decrease in [Ca(2+)](i) during thrombin-mediated platelet activation. The functional consequence of reduced PMCA tyrosine phosphorylation and decreased [Ca(2+)](i) was a significant delay in the onset of thrombin-mediated platelet aggregation. CONCLUSIONS: The results demonstrate that PMCA tyrosine phosphorylation regulates [Ca(2+)](i) during platelet activation, which affects downstream events in the activation process. Moreover, PMCA tyrosine phosphorylation and resultant inhibition of PMCA activity produces a positive feedback loop mechanism by enhancing the increase in [Ca(2+)](i) accompanying platelet activation.     

Spectral Turbulence Versus Time-Domain Analysis of Signal-Averaged ECG Used for the Prediction of Different Arrhythmic Events in Survivors of Acute Myocardial Infarction

Spectral Turbulence SAECG After MI. Introduction : Spectral turbulence analysis of the signal-averaged ECG (SAECG) combines spectral analysis with statistical evaluation of spectrograms of individual parts of the QRS complex. It has been suggested that it may be superior to conventional time-domain analysis of the SAECG.
Methods and Results : This study compared the power of conventional time-domain (40 to 250Hz) and spectral turbulence analyses of SAECG for the prediction of cardiac death, ventricular tachycardia, sudden arrhythmic death, and arrhythmic events (ventricular tachycardia or fibrillation, and/or sudden arrhythmic death) after acute myocardial infarction in 603 patients. The population excluded patients with bundle branch block and other conduction abnormalities. During the first 2 years of follow-up, there were 40 cardiac deaths, 21 cases of ventricular tachycardia, 11 sudden arrhythmic deaths, and 29 arrhythmic events. The positive predictive accuracy of spectral turbulence analysis was significantly higher than time-domain analysis for cardiac death at most levels of sensitivity (e.g., 26% vs 20% at 40% sensitivity, P < 0.05). The positive predictive accuracies of the two techniques were not statistically different for the prediction of ventricular tachycardia. For the prediction of sudden arrhythmic death and arrhythmic events, the positive predictive accuracy of spectral turbulence was better than that of time-domain analysis only at the higher levels of sensitivity (9% vs 2%, P < 0.001 for sudden arrhythmic death at 60% sensitivity, and 14% vs 11%, P < 0.05 for arrhythmic events at 60% sensitivity).
Conclusions : Spectral turbulence analysis is essentially equivalent to time-domain analysis for the prediction of arrhythmic events after myocardial infarction. However, it performed significantly better than time-domain analysis for the prediction of cardiac death.     

Statistical Distribution of Ventricular Ectopic Beats Assessed by Computer Analysis of Long-Term Electrocardiograms: Problems Related to the Definition of Arrhythmogenesis

The severity and clinical importance of arrhythmogenic effects of antiarrhythmic drugs have been well recognized, but only empirical and naive approaches have been used to define antiarrhythmic and proarrhythmic effects when comparing baseline and on-therapy Holter recordings. This article suggests a new method for the assessment of these effects, based upon comparisons of statistical distributions of ectopic beats recorded on long-term electrocardiograms. Since these distributions are very individual, the Smirov test was used for their comparison. This enables the antiarrhythmic and arrhythmogenic effects to be defined on a precise statistical basis. To demonstrate this method, an analysis is reported of Holter recordings made on six patients suffering from different types of ventricular arrhythmia. In each of these patients, one baseline Holter recording and one recording on each of three different drugs (flecainide, sotalol, and verapamil) were made. The records were digitized using a commercially available system for the analysis of long-term electrocardiograms and random sampling was used to obtain independent samples of arrhythmic episodes that were subsequently statistically analyzed. The results show that the problem of the definition of arrhythmogenesis can be partly addressed in a precise mathematical way and that treatment with an antiarrhythmic preparation may not only have general antiarrhythmic or arrhythmogenic effects, but may also cause a significant change in the character of arrhythmia that can neither be classified as antiarrhythmic nor as arrhythmogenic influence. The clinical implications of this approach are discussed.     

Systematic Comparisons of Electrocardiographic Morphology Increase the Precision of QT Interval Measurement

Decreased intrasubject variability of QTc values is needed to increase the power and reduce the size of the so-called thorough QT studies. One source of QTc variability is the lack of systematic measurements when electrocardiograms (ECG) with closely matching morphologies are not measured in an exactly corresponding way. The inaccuracy can be eliminated by postprocessing of QT measurements by ECG pattern matching. This study tested the effects of pattern matching in ECG measurements in two populations of healthy subjects (n = 48 + 56) and in a population of patients with advanced Parkinson's disease (n = 130) in whom both day-time and night-time data were available. Intrasubject QTc variability was measured by intrasubject standard deviations (SD) of QTc values obtained with manual measurements before and after pattern-matching measurement alignments. In each subject, QT values (n = 230–320) in one drug-free long-term ECG recording were evaluated. The pattern-matching adjustment of the QT measurement decreased the intrasubject QTc variability from 5.2 ± 1.0 to 4.5 ± 1.0 ms (P < 10⁻¹⁴) from 6.4 ± 1.7 to 5.5 ± 1.6 ms (P < 10⁻¹⁰) from 5.6 ± 1.5 to 4.6 ± 1.4 ms (P < 10⁻³⁴) and from 6.1 ± 1.9 to 5.0 ± 1.7 ms (P < 10⁻³³), in the two populations of healthy subjects and in the day-time and night-time recordings of Parkinson's disease patients, respectively. Hence, morphological pattern adjustment of QT interval measurements improves the quality of the QT data with substantial practical implications. Reductions in intrasubject QTc variability were reproducibly found in different populations and thus the technology might be recommended for every thorough QT/QTc study. Noticeable reductions of necessary study size are likely achievable in this way.     

APOPTOSIS OF HUMAN MONOCYTES/MACROPHAGES IN MYCOBACTERIUM TUBERCULOSIS INFECTION

Tuberculosis (TB) is still a major health problem, both as a single disease entity and as a cofactor in AIDS. The interaction between macrophage and Mycobacterium tuberculosis (MTB) is a critical step in the establishment of an early chronic infection. This study analyses the capacity of MTB to induce apoptosis in cells obtained by broncho-alveolar lavage (BAL) from patients with reactive pulmonary tuberculosis and from AIDS patients with disseminated pulmonary tuberculosis. Apoptosis was increased three-fold in BAL cells obtained from patients with pulmonary tuberculosis and even more markedly in alveolar macrophages of MTB-infected AIDS patients, compared with controls. Apoptosis was analysed and characterized by propidium iodide (PI) incorporation, terminal deoxy transferase (TDT)-mediated dUTP-biotin nick end labelling (TUNEL), and tissue transglutaminase (tTG) expression. The MTB–macrophage interaction was also investigated in vitro by infecting monocyte-derived macrophages (MDM) with MTB (virulent strain H37Rv). The induction of apoptosis by MTB required viable bacteria, was dose-dependent, and was restricted to H37Rv. Infection with either Mycobacterium avium complex (MAC) or HIV-1 and treatment with heat-killed MTB failed to induce apoptosis. © 1997 by John Wiley & Sons, Ltd.     

Comparison of Formulae for Heart Rate Correction of QT Interval in Exercise Electrocardiograms

The study investigated the differences in five different formulae for heart rate correction of the QT interval in serial electrocardiograms recorded in healthy subjects subjected to graded exercise. Twenty-one healthy subjects (aged 37+/-10 years, 15 male) were subjected to graded physical exercise on a braked bicycle ergometer until the heart rate reached 120 beats/min. Digital electrocardiograms (ECG) were recorded on baseline and every 30 seconds during the exercise. In each ECG, heart rate and QT interval were measured automatically (QT Guard package, Marquette Medical Systems, Milwaukee, WI, USA). Bazett, Fridericia, Hodges, Framingham, and nomogram formulae were used to obtain QTc interval values for each ECG. For each formula, the slope of the regression line between RR and QTc values was obtained in each subject. The mean values of the slopes were tested by a one-sample t-test and the comparison of the baseline and peak exercise QTc values was performed using paired t-test. Bazett, Hodges, and nomogram formulae led to significant prolongation of QTc intervals with exercise, while the Framingham formula led to significant shortening of QTc intervals with exercise. The differences obtained with the Fridericia formula were not statistically significant. The study shows that the practical meaning of QT, interval measurements depends on the correction formula used. In studies investigating repolarization changes (e.g., due to a new drug), the use of an ad-hoc selected heart rate correction formula is highly inappropriate because it may bias the results in either direction.     

Changes in Heart Rate Variability with Age

Depressed heart rate variability (HRV) after a myocardial infarction is associated with increased mortality. This is thought to be due to reduced parasympathetic activity and heightened sympathetic activity. Aging is associated with depressed HRV, but little is known of the affect of aging on parasympathetic activity. This study examined 56 healthy subjects (age range 40–102 years; 39 women). None had a history of heart disease or were on medication that would affect cardiac function. All had normal resting ECGs, normal heart size on chest X ray, and normal electrolytes. In all subjects, 24-hour Holter recordings were performed and used to measure HRV. In particular, the study examined the affect of age on HRV triangular index, which gives an estimate of overall HRV, and on RMSSD (square root of the mean squared differences of successive normal-to-normal RR intervals), which gives an estimate of short-term components of HRV and is thought to reflect the overall extent of vagal modulations of heart rates. Both these parameters were compared in patients younger and older than 70 years. Each recording lasted at least 17 hours; the majority of recordings were longer than 20 hours. There was a significant decrease in HRV triangular index with age (r =?0.4, P < 0.05) and no significant change in RMSSD with age(r =?0.08, P = NS). There was a significant difference in HRV index in those > 70 years compared with those < 70 years (38.0 ± 9.3 vs 31.0 ± 11, respectively, P <0.02). There was no significant difference in RMSSD between the two age groups (26.7 ± 8.2 ms vs 28.4 ± 11.3 ms, respectively, P = NS). Thus, the study concludes that aging reduces the global measure of HRV and may reflect reduced responsiveness of autonomic activity to external environmental stimuli with age. However, the time-domain short-term components of HRV are not affected by age and, therefore, the fast and presumably vagal modulations of heart rate appear to be maintained.