D. Eugene Lovelace

Secondary structure of detergent-solubilized phospholamban, a phosphorylatable, oligomeric protein of cardiac sarcoplasmic reticulum

The structure of phospholamban, a 30-kDa oligomeric protein integral to cardiac sarcoplasmic reticulum, was probed using ultraviolet absorbance and circular dichroism spectroscopy. Purified phospholamban was examined in three detergents: octyl glucoside, n-dodecyloctaethylene glycol monoether (C12E8) and sodium dodecyl sulfate (SDS). Ultraviolet absorption spectra of phospholamban reflected its aromatic amino acid content: absorption peaks at 275-277 nm and 253, 259, 265 and 268 nm were attributed to phospholambans one tyrosine and two phenylalanines, respectively. Phospholamban phosphorylated at serine 16 by the catalytic subunit of cAMP-dependent protein kinase exhibited no absorbance changes when examined in C12E8 or SDS. Circular dichroism spectroscopy at 250-190 nm demonstrated that phospholamban possesses a very high content of alpha-helix in all three detergents and is unusually resistant to denaturation. Dissociation of phospholamban subunits by boiling in SDS increased the helical content, suggesting that the highly ordered structure is not dependent upon oligomeric interactions. The purified COOH-terminal tryptic fragment of phospholamban, containing residues 26-52 and comprising the hydrophobic, putative membrane-spanning domain, also exhibited a circular dichroism spectrum characteristic of alpha-helix. Circular dichroism spectra of phosphorylated and dephosphorylated phospholamban were very similar, indicating that phosphorylation does not alter phospholamban secondary structure significantly. The results are consistent with a two-domain model of phospholamban in which each domain contains a helix and phosphorylation may act to rotate one domain relative to the other.

Echocardiographic detection of ischemic and infarcted myocardium

The purpose of this study was to determine the potential of a clinically adaptable two-dimensional echocardiographic system using computer enhancement and a mathematically defined integrated backscatter ratio for the early detection of ischemic and infarcted myocardium. Fifteen dogs had two-dimensional echocardiograms recorded during either open chest coronary occlusion (n = 5), closed chest occlusion (n = 5), occlusion followed by reperfusion (n = 3) or sham coronary occlusion (n = 2). A serial increase in integrated backscatter ratio, representing differences in returned ultrasound intensities between a reference point and specific myocardial regions, was detected between 7 and 12 minutes of complete occlusion in 9 of 12 animals (p less than 0.05), and at minutes 18, 43 and 67 in the remaining 3 animals. Reperfusion after 20 minutes of occlusion in two studies resulted in normalization of the backscatter ratio. An increase in backscatter ratio was not detected when 5 minute occlusion periods were used or during the 5 hour sham occlusion studies. The computer enhancement techniques utilized in this study provided increased visual detail of intracardiac structures over that provided by routine two-dimensional echocardiograms; myocardial tissue was identifiable in what appeared to be echo-free segments; and boundaries that appeared as noncontiguous horizontal lines on the routine echocardiograms were identifiable as trabeculae. The results indicate that: 1) significant increases in backscatter from nonperfused myocardium are detectable echocardiographically within 12 minutes of coronary occlusion and temporal changes can be assessed in the canine model, and 2) the echocardiographic data acquisition and computer analysis system utilized provide a clinically adaptable approach to identify and map myocardial characteristics in human beings.

Computer quantitation of Q-T and terminal T wave (aT-eT) intervals during exercise: Methodology and results in normal men

Computer-quantitated measurements of the Q-T intervals, the Q-T/Q-Tc ratio (Q-T/corrected Q-T) and the terminal T wave (apex to end of T [aT-eT] interval) were evaluated in resting and exercise electrocardiograms of 130 normal men with a mean age of 40 years. Pseudo-orthogonal, bipolar X, Y and Z axis leads were recorded during treadmill exercise testing, and 25 consecutive QRS-T complexes from standing rest and three exercise stages were computer-averaged. The Q-T intervals, Q-T/Q-Tc ratio and aT-eT interval measurements were then computed in the X and Z axis leads only, because the Y lead proved to be too noisy for accurate interpretation. A correlation coefficient of 0.9830 resulted between measurements made manually from the plotted, composite QRS-T complexes and those made by computer. No significant differences , in the paired sense, were found between any of the measurements. Measurements made on the Z axis lead; however, the differences in the measurements remained constant across all stages of exercise. A Q-T/Q-Tc ratio of greater than 1.08, previously reported to be a reliable indicator of coronary disease, was observed in the majority of our normal subjects during exercise. Although the Q-T interval is substantially influenced by many factors, the aT-eT interval proved not to be age- or heart rate-dependent. It appears that the aT-eT interval can be measured with a high degree of reliability during exercise and it may prove to be a relatively specific indicator of repolarization alterations that occur with myocardial ischemia.

Behavior of the terminal T wave during exercise in normal subjects, patients with symptomatic coronary artery disease and apparently healthy subjects with abnormal ST segment depression

The Q-T interval and apex of T wave to end of T wave (aT-eT) interval were measured by computer in four age-matched study groups at rest and during exercise to determine whether: the behavior of the aT-eT interval differs in patients with myocardial ischemia when compared with normal subjects, and the behavior of the aT-eT interval differs in subjects with true positive and false positive ST segment responses. Group I consisted of 57 normal subjects. Group II consisted of 41 symptomatic patients with documented coronary artery disease. A group of apparently healthy subjects with asymptomatic ST segment depression during exercise was divided into two additional groups: Group III, those without coronary artery disease; and Group IV, those with coronary artery disease. Subjects were excluded from the study if they had left ventricular hypertrophy or an intraventricular conduction defect or were taking digitalis or type I antiarrhythmic agents. There were no significant differences in the aT-eT interval and aT-eT/Q-T ratio among the four study groups when compared at rest; however, during exercise at similar heart rates, the aT-eT interval was significantly shorter and the aT-eT/Q-T ratio significantly smaller in Groups II and IV, the subjects with coronary artery disease, than in Group I, the normal subjects. The aT-eT interval and aT-eT/Q-T ratio measurements in Group III did not differ from those in Group I at rest or during exercise. In conclusion, the aT-eT interval and aT-eT/Q-T ratio may reflect changes in myocardial repolarization in exercise-induced ischemia and may have potential for future clinical application.

A computerized cardiovascular data base system: Application in arrhythmia research

Abstract A minicomputer-based data management system for the storage, retrieval, and analysis of cardiovascular clinical and basic research data is described and examples given of its potential utility in clinical arrhythmia research. The system links multiple data bases so that subsetting, statistical analyses and a variety of other data manipulations can be accomplished across data bases. Data additions or refinements can be made without restructuring an entire data base and new questions asked without the need for specific program revisions. The system is user-oriented. All aspects from data formulation through analysis can be accomplished by clinical investigators without computer or mathematical backgrounds.

Nonparoxysmal junctional tachycardia in acute myocardial infarction: Computer-assisted detection

Thirty consecutive patients with acture myocardial infarction had continuous magnetic tape recording of their stay in the coronary care unit. Analysis of the 24 hour tape recordings was implemented on a Honeywell model 316 digital computer. In the first 24 hours after admission to the coronary care unit, 12 of the 30 patients (40 percent) exhibited nonparoxysmal junctional tachycardia; in 5 the arrhythmia was not recognized by conventional monitoring techniques. For the subsequent 3 days, the incidence rate of the arrhythmia was 13 percent for the first 48 hours and 3 percent for 72 hours. Although the mortality rate in patients with nonparoxysmal junctional tachycardia was greater than in patients not demonstrating the arrhythmia (33 versus 6 percent), there was a greater percentage of patients with anterior infarction in the former group; therefore, mortality may have been related to site of infarction rather than being reflective of the arrhythmia or its associated pathophysiologic state. Of possible significance is the association of a greater degree of sinus arrhythmia with nonparoxysmal junctional tachycardia. The incidence of nonparoxysmal junctional tachycardia in this group of patients was greater than previously reported. It is possible that computer tape analysis may have provided more accurate recognition of the arrhythmia and, thus, more realistic incidence data. The association of nonparoxysmal junctional tachycardia with sinus arrhythmia could only have been recognized by computer technique. The computer system is not a diagnostic system but rather a tape review method.

The phase-plane cardiogram: preliminary findings on a healthy population

Summary A method for increasing the diagnostic capability of the clinical electrocardiogram has been further developed. The coordinated display of voltage against the time derivative of voltage (dv/dt), i.e., PPC, was found to be remarkably sensitive to subtle aberrations in QRS contours not easily visualized in the standard electrocardiographic portrayal of voltage against time. A standard twelve-lead electrocardiogram and phase-plane loops were displayed and photographed on an oscillographic recorder, the latter by placing voltage (V) on the vertical axis and the first time derivative of voltage on the horizontal axis. Data storage on magnetic tape and computer analysis of the data were also carried out. Data from 100 “normal” (as determined from clinical ECG, history, and physical examination) subjects demonstrated evident, repeatable notching patterns in the PPC. The repeatability of these patterns was demonstrated from beat-to-beat in each lead of every individual, and was found to be consistent even when the readings were taken over long periods of time. The PPCs of different individuals showed consistent common notching patterns which would indicate that anatomical and physiological bases exist to explain this phenomenon.