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The sonication of human serum albumin produces air-filled microspheres that are used in echocardiographic studies of myocardial perfusion. Recent studies suggest that the microspheres disappear when high pressures are applied, altering the relationship between the administered microsphere dose and the echocardiographic response. Because an ultrasound pulse generates a pressure wave in insonified medium, we hypothesized that with increasing acoustic pulse pressure, the microsphere concentration decreases, hence ultrasonic backscatter decreases. We measured relative integrated backscatter from albumin microspheres diluted in normal saline solution (6152 microspheres/ml) and 5% human plasma protein fraction (24,608 microspheres/ml), with increasing acoustic pulse pressures at the transducer's focus. Backscatter was also measured in normal saline solution with increasing concentrations (up to 15,380 microspheres/ml) of albumin microspheres at an acoustic pulse pressure of 0.11 MPa (1.1 atm). Backscatter and microsphere concentration were related logarithmically: y = 3.38 x 0.32; r = 0.93. Backscatter was unchanged over time at acoustic pulse (peak compression) pressures less than 0.15 MPa (1.5 atm). However, backscatter decreased readily at acoustic pulse pressures greater than 0.33 MPa (3.3 atm), which included any mixing effects. Thus, albumin microspheres are acoustically labile.

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BACKGROUND: Although transesophageal echocardiography (TEE) produces real-time images depicting left ventricular (LV) filling and ejection, the quantitative analysis of these images has been too time consuming to be of practical value in the operating room. Therefore, the authors investigated whether a new automated border detection system (ABD) could track the endocardial border continuously and compute the cross-sectional area of the LV cavity. METHODS: Using data from 25 patients who were monitored with TEE as part of their routine clinical care, the authors compared ABD estimates of LV end-diastolic area (EDA in square centimeters), end-systolic area (ESA in square centimeters), and fractional area change (FAC) with the laboratory measurements made independently by an expert. RESULTS: ABD slightly underestimated EDA (10.7 +/- 1.0 vs. 11.2 +/- 1.0 cm2) and slightly overestimated ESA (5.6 +/- 0.7 vs. 4.8 +/- 0.6 cm2, mean +/- standard error). However, when ABD tracking of the endocardial border was judged as "good" or "excellent" (84% of the patients at end diastole and 72% at end systole), the limits of agreement between ABD and the expert's findings were within the limits expected for two experts. By contrast, ABD significantly underestimated FAC (0.44 +/- 0.03 vs. 0.56 +/- 0.03) and the limits of agreement between ABD and the expert were more than twice as great as expected for experts, even when ABD performance was judged as "excellent." CONCLUSION: The authors conclude that, when ABD appears to be performing adequately, it underestimates LV FAC, but provides valid real-time estimates of LV EDA and ESA. Thus, it warrants further evaluation as a potentially powerful clinical and research tool.

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BACKGROUND: Automated edge detection of endocardial borders in echocardiograms provides objective, reproducible estimation of cavity area; however, most methods have required off-line analysis. A recently developed prototype echocardiographic imaging system permits real-time automated edge detection during imaging and thus, the potential for measurement of cyclic changes in cavity area and the assessment of left ventricular function on-line. Our purpose was to compare measurements of endocardial area manually traced from conventional echocardiograms with those obtained with the real-time automated edge detection system in normal subjects. METHODS AND RESULTS: Two training sets of images were used to establish optimal methods of gain setting; the settings were then evaluated in a test set of images. In the high-gain training group (n = 8 subjects, 119 images), gain settings were adjusted sufficiently high to display at least 90% of the endocardial border. Manually drawn and real-time area measurements correlated at r = 0.92, but manually drawn areas were underestimated by computer. In the low-gain training group (n = 7 subjects, 104 images), gain settings were adjusted sufficiently low to avoid cavity clutter despite the presence of dropout of endocardial edges. Manually drawn and real-time areas again correlated (r = 0.79), but manually drawn areas were overestimated by computer. In the intermediate-gain test group (n = 7 subjects, 105 images), gain settings were balanced between maximal endocardial definition (greater than or equal to 90%) and minimal cavity clutter (less than or equal to 1 cm2). Manually drawn and real-time areas correlated at r = 0.91 for the group, and r ranged from 0.94 to 0.99 in individual subjects. Interobserver variability was 9.5% for manually traced areas and 10.6% for real-time area measurements. CONCLUSIONS: Real-time on-line automated edge detection provides accurate estimation of manually drawn cavity areas. Although the method is gain dependent, measurements are reproducible. The system should have clinical application in settings in which measurements of left ventricular function are important.

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Cyclic backscatter variation is useful in differentiating normal from ischemic and myopathic myocardium; however, there are few data on the reproducibility of clinical cyclic variation measurements. Therefore, a study using 2-dimensional and M-mode backscatter imaging was performed in 20 normal male subjects by 2 observers at an initial session and by 1 of the observers after 1 week. Cyclic variation on M-mode was calculated as the difference between the end-diastolic backscatter and the backscatter at the nadir. Two-dimensional determinations of backscatter were made using a single frame at end-diastole and one at end-systole. The cyclic change was the difference between backscatter measured in the end-diastolic and end-systolic frames. There were no statistically significant differences in analysis of variance among the grouped repeated measurements in either the interventricular septum or the posterior left ventricular wall. At the initial session, cyclic backscatter variation in the posterior wall using M-mode techniques was 5.9 +/- 1.8 dB (SD). The cyclic change in backscatter in the septal wall, using the 2-dimensional technique, was 4.3 +/- 2.4 dB. In the posterior wall, the cyclic change in backscatter was 5.7 +/- 1.7 dB. Pairwise observer correlations between repeated measurements ranged from -0.48 to 0.45. Thus, although there were no significant differences in group means on repeat measurements, repeated measurements in individual subjects were not reliably reproduced because of limited independent sampling of backscatter measurements at only 2 points in the heart cycle. Increased independent sampling and measurement from a backscatter waveform throughout the cardiac cycle may improve reproducibility of measurements.

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To provide an approach suitable for on-line analysis of ventricular function, a conventional two-dimensional ultrasound imaging system was modified to detect and track blood-tissue interfaces in real time based on their quantitative acoustic properties. This modification permitted on-line display of the left ventricular cavity area, fractional area change, volumes and ejection fraction on a beat by beat basis. Images were obtained from 54 patients and 12 normal subjects with broad ranges of ventricular dimensions and systolic function. On-line measurements of cavity areas were compared with off-line measurements of cavity areas (analysis of videotaped conventional images). Left ventricular cavity areas measured on-line from short-axis views correlated closely with off-line views as did areas from apical views. On-line fractional area change correlated well with ejection fraction calculated off-line. More than 70% of patients could be studied adequately with the approach developed. Thus, automatic boundary detection based on quantitative assessment of tissue acoustic properties permits on-line quantitation of ventricular cavity areas and indexes of function.

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Acute myocardial ischemia and chronic myocardial infarction may be recognized with ultrasound tissue characterization techniques because of myocardial acoustic changes caused by reduced perfusion and/or collagen deposition. Our purpose was to study the acoustic properties of recent myocardial infarction when the predominating pathologic finding was myocardial edema and leukocytic infiltration. We used a new quantitative backscatter imaging system to study 18 patients 9 +/- 5 days after myocardial infarction (eight patients with anteroseptal myocardial infarction and 10 with inferior myocardial infarction) and 20 normal subjects. The cyclic variation of relative integrated backscatter (end-diastolic minus end-systolic) was calculated from on-line measurements. Standard parasternal long- and short-axis and apical four- and two-chamber views were obtained. In the anteroseptal myocardial infarction group, the cyclic variation of relative integrated backscatter was lower in the septum (1.5 +/- 1.6 dB) than in the posteroinferior wall (3.2 +/- 1.2 dB); however, the sample size of only three patients (of eight patients imaged) in the latter group prevented statistical comparison. The cyclic variation of relative integrated backscatter in the infarcted septum was less than the measurement obtained in the septum of the control group (4.3 +/- 2.4 dB, p less than 0.05). In the inferior infarction group, the cyclic variation of integrated backscatter in the posteroinferior wall (1.8 +/- 1.7 dB) was not significantly different from the measurement obtained in the septum (3.7 +/- 3.6 dB); however, the cyclic variation in the posteroinferior wall was significantly less than that obtained in the control group posteroinferior wall (5.7 +/- 1.7 dB, p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

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Two-dimensional echocardiography is an excellent technique for detecting left ventricular thrombi, however, acute clot is sometimes difficult to differentiate from adjacent myocardium and intracavitary signals. We hypothesized that quantitative assessment of the acoustic properties of acute left ventricular thrombi using a quantitative backscatter imaging system would permit the differentiation of thrombus from adjacent myocardium and intracavitary echoes. Acute, experimental left ventricular thrombi in seven dogs were evaluated with a quantitative backscatter imaging system that allowed the measurement of relative integrated backscatter and cyclic (i.e., diastolic minus systolic) variation in integrated backscatter. Coronary ligation abolished the cyclic variation in relative backscatter that occurred in normal myocardium. The end-diastolic relative backscatter in the thrombus (16.9 +/- 1.3 dB) was significantly higher than in apical myocardium (13.2 +/- 0.6 dB, p less than 0.05). There was no significant difference in the cyclic variation in relative backscatter among thrombus, ischemic myocardium, or intracavitary blood. Thus, the quantitative assessment of the acoustic properties of left ventricular thrombi can be useful in their detection and in the differentiation from myocardium and intracavitary signals.

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We have shown previously that the physical properties of myocardium in dogs can be characterized with quantitative ultrasonic integrated backscatter and that interrogation of the tissue with ultrasound can delineate cardiac cycle-dependent changes in ultrasonic backscatter in normal tissue that disappear with ischemia and reappear with reperfusion if functional integrity is restorable. To determine whether this approach can be applied to man, we implemented an automatic gain compensation and continuous data acquisition system to characterize myocardium with quantitative ultrasonic backscatter and to detect cardiac cycle-dependent changes in real time. We developed a two-dimensional echocardiographic system with quantitative integrated backscatter imaging capabilities for use in human subjects that can automatically differentiate ultrasonic signals from blood as opposed to those obtained from tissue and adjust the slope of the gain compensation appropriately. Real-time images were formed from a continuous signal proportional to the logarithm of the integrated backscatter along each A-line. In our initial investigation, 15 normal volunteers (ages 17 to 40 years, heart rates 44 to 88 beats/min) and five patients with dilated cardiomyopathy (ages 22 to 52, heart rates 82 to 120 beats/min) were studied with conventional parasternal long-axis echocardiographic views. Diastolic-to-systolic variation of integrated backscatter in the interventricular septum and left ventricular posterior wall was seen in each of the normal subjects averaging 4.6 +/- 1.4 dB (SD) and 5.3 +/- 1.5 dB (n = 127 sites), respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

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A simple model for predicting the degree of speckle reduction in A-mode images via the correlation coefficient was tested experimentally. The model appeared to account correctly for the influence of image system bandwidth on frequency separations required for nearly independent sampling in compound frequency imaging. Moreover, the notion of compound bandwidth imaging was tested; the correlation between samples at different bandwidths decreased as the ratio of bandwidths decreased.

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Integrated ultrasonic backscatter has been related to collagen deposition in fibrotic myocardium. The purpose of our study was to measure the integrated ultrasonic backscatter in the right and left ventricles of 10 normal freshly excised canine hearts and five normal formalin-fixed human hearts. A 2.25 MHz, 50% fractional bandwidth transducer was positioned at the transducer focal distance from the epicardium. The radio frequency backscatter signal, excluding specular reflections, was digitized, squared, and integrated to yield the integrated ultrasonic backscatter (in decibels down from a 100% reflector). The segment of myocardium corresponding to the integrated ultrasonic backscatter sample volume was excised and assayed for hydroxyproline, a marker for collagen. A second purpose of our study was to evaluate the influence of fixation with formalin on the backscatter. Regional integrated ultrasonic backscatter was therefore measured in 10 freshly excised canine left ventricles, which were fixed in 10% formalin for 2 weeks. Integrated ultrasonic backscatter measurements were then repeated. In freshly excised canine hearts, the integrated ultrasonic backscatter from right ventricle was higher than that from left ventricle (-60.4 +/- 1.6 [SEM] vs -66.9 +/- 1.0 dB; p less than .001). The collagen content of right ventricle was also higher than that of left ventricle (4.40 +/- 0.26 [SEM] vs 3.58 +/- 0.13 micrograms/mg dry weight; p less than .005). Similar results were obtained in human hearts. There were no correlations between integrated ultrasonic backscatter and collagen content (r = .28 and .32 for dogs and humans, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

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Regional two-dimensional (2D) echocardiographic amplitude patterns, or image texture, may be of diagnostic importance. Echocardiographic image texture is due in part to acoustic speckle, a complex pattern of interference of reflections from many small scatterers in tissue. The regional speckle pattern appears to be altered in several disorders associated with abnormalities in myocardial structure but also may be altered by a variety of characteristics of the scanning instrument. We hypothesized that quantitative measures of regional 2D echocardiographic image texture would vary as a function of position in range and azimuth within the field of view, even when imaging a uniform ensemble of scatterers. We tested this hypothesis by imaging a tissue-equivalent phantom with two phased-array scanners and two different methods of digitization. We analyzed the texture in several regions of interest separated in range and azimuth and found significant differences in quantitative texture measures as a function of position of the region of interest in the sector field of view (p values .006 to .0001 by multivariate analysis of variance). We found significant regional variability in texture with both scanners and both methods of digitization. We conclude that regional quantitative image texture in 2D echocardiograms varies as a function of range and azimuth, even when imaging a uniform ensemble of scatterers. This variability is related to several physical and instrument-related phenomena and precludes interpreting all regional texture alterations as indicating tissue structural abnormalities.

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Previous approaches to automatic border recognition in two-dimensional echocardiography have utilized off-line computer image analysis. In this paper, we describe a hardware technique for real-time endocardial edge detection during the echocardiographic examination. This technique is based on a new method of line-by-line compensation for ultrasound attenuation. The detected edges and composite edge images can be displayed alone or superimposed upon the usual echocardiographic display. To test the accuracy of our real-time technique, we obtained echocardiograms of excised hearts and compared real-time edges with edges manually traced by an experienced observer. Left ventricular cavity areas and endocardial perimeters derived from real-time edges correlated well with those derived from observer-defined edges.

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A new method of compensating for the attenuation of ultrasound by chest wall, myocardium and blood has been devised by using an optimum threshold to distinguish between blood and myocardium. This allows changing receiver gain at rates suitable for either blood or myocardium for each line-of-sight forming a real-time image. Compensation artifacts arising from the effects of speckle or phase cancellation are minimized by using a one-bit digital filter after threshold detection. This method of compensation is a necessary first step toward quantitative two-dimensional echocardiography. ?NHLBI 14388

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We hypothesized that acute myocardial infarction could be detected in standard two-dimensional echocardiograms of closed-chest dogs by evaluating regional echo amplitude distributions using computerized image analysis. We tested this hypothesis by performing standard, 2.4 MHz two-dimensional echoes before and 2 days after circumflex coronary occlusion in seven closed-chest dogs. Control and infarcted regions of interest were studied in digitized stop-frame images. Average gray level was calculated for each region of interest, and the shape of the gray-level distribution was analyzed by calculation of skewness and kurtosis and by qualitative features of shape. Average gray level increased significantly from the pre- to postocclusion images in the infarcted regions (16.7 +/- 4.2 vs. 32.4 +/- 4.4 units, P less than 0.01), but not in the control regions (17.4 +/- 4 vs. 22.3 +/- 5.5., P = NS). Average gray level could not distinguish between infarcted and normal regions within the postocclusion images (36 +/- 5.2 vs. 33.6 +/- 5.8, P = NS). Three independent observers qualitatively evaluated histogram shape and correctly identified 7/7 MI regions (100% sensitivity) and 14/20 normal regions (70% specificity). Quantitatively, infarct regions exhibited a significant decrease in kurtosis from pre- to postocclusion images (7.1 +/- 4.0 vs. 5.2 +/- 2.9, P = NS). Within postocclusion images, infarcted regions displayed a significantly lower kurtosis than did normal regions (0.27 +/- .47 vs. 2.5 +/- 1.0, P less than .01). We conclude that acute myocardial infarction may be detected in closed-chest dogs by analyzing regional echo amplitude data from standard two-dimensional echocardiograms.

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A simple assay based on molecular light scattering was developed to detect soluble immune complexes. In this assay the percentage of relative light scattering (%RLS) caused by complexes in buffer containing polyethylene glycol 6000 (PEG) was measured with a laser nephelometer. In low concentrations of PEG (3%-5%), IgG-anti-IgG complexes formed in antigen excess scattered light more effectively than those formed in antibody excess or equivalence. Since normal serum proteins caused minimal %RLS at 3%-5% PEG, we expanded this assay to screen patient sera. For each test, the serum was used at a 1:20 dilution. The results were expressed by the defined parameter delta %RLS = (sigma %RLS 3%-5% PEG - sigma %RLS 0%-2% PRG). For 58 normal sera delta %RLS = 15.4% +/- 11.3%, while for 12 sera from patients who had systemic lupus erythematosus and hypocomplementemia, delta %RLS = 40.5% +/- 20.0% (P < 0.001). In a comparative study of 18 sera positive for immune complexes in the Clq deviation assay, 12 were positive by %RLS.

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