Three-dimensional characterization of human ventricular myofiber architecture by ultrasonic backscatter.

Normal human left ventricular architecture comprises a highly aligned array of cardiac myofibers whose orientation depends on transmural location. This study was designed to determine whether measurement of integrated backscatter could be used detect the progressive transmural shift of myofiber alignment that occurs from epicardium to endocardium in human ventricular wall segments. Integrated backscatter was measured at 32 transmural levels in seven cylindrical biopsy specimens (1.4 cm diam) sampled from normal regions of six explanted fixed human hearts by insonification of samples at 180 independent angles in 2 degrees steps around their entire circumference with a 5-MHz broadband piezoelectric transducer. Histologic analysis was performed to determine fiber orientation. Integrated backscatter varied approximately as a sinusoidal function of the angle of insonification at each transmural level. Greater integrated backscatter was observed for insonification perpendicular as compared with parallel to fibers (difference = 14.5 +/- 0.6 dB). Ultrasonic analysis revealed a progressive transmural shift in fiber orientation of approximately 9.2 +/- 0.7 degrees/mm of tissue. Histologic analysis revealed a concordant shift in fiber orientation of 7.9 +/- 0.8 degrees/mm of tissue. Thus, human myocardium manifests anisotropy of ultrasonic scattering that may be useful for characterization of the intramural fiber alignment and overall three-dimensional organization of cardiac myofibers.     

Three-dimensional visualization and thickness estimation of aortic valve cusps using high-frequency ultrasound

High-frequency ultrasound techniques are introduced for three-dimensional imaging and thickness estimation of fresh heart valve cusps. Images of porcine aortic valve specimens were acquired within a 12 x 8 x 8 mm3 volume using a VisualSonics VS40 micro-imaging system operating at a 40 MHz centre frequency. Two image volumes were obtained from each of six left coronary cusps. One volume was acquired with the specimen submerged in distilled water and the second volume was acquired through either Hanks physiologic solution or coronary perfusion solution (CPS). The fibrosa, spongiosa and ventricularis were most readily distinguished when the specimen was imaged in distilled water. Colour thickness maps were computed from B-mode image data, and the mean and standard deviations of the thickness were determined for each cusp. In 11 of 12 trials, the image analysis algorithm yielded valid thickness estimates over greater than 98% of the region examined. Mean thickness estimates obtained with specimens submerged in Hanks solution or CPS ranged from 0.66 to 1.03 mm, and submersion in distilled water increased the mean thickness by 20-40%. This observation suggests that the cusps osmotically absorbed water. Information provided by high-frequency ultrasound is expected be valuable for characterizing the morphological properties of heart valves.     

Quantitative assessment of aortic sclerosis using ultrasonic backscatter.

BACKGROUND: The development of therapeutic interventions to prevent progressive valve damage is more likely to limit the progression of structural damage to the aortic valve with normal function (aortic sclerosis [ASC]) than clinically apparent aortic stenosis. Currently, the ability to appreciate the progression of ASC is compromised by the subjective and qualitative evaluation of sclerosis severity. METHODS: We sought to reveal whether the intensity of ultrasonic backscatter could be used to quantify sclerosis severity in 26 patients with ASC and 23 healthy young adults. Images of the aortic valve were obtained in the parasternal long-axis view and saved in raw data format. Six square-shaped 11 x 11 pixel regions of interest were placed on the anterior and posterior leaflets, and calibrated backscatter values were obtained by subtracting the regions of interest in the blood pool from the averaged backscatter values obtained from the leaflets. RESULTS: Mean ultrasonic backscatter values for sclerotic valves exceeded the results in normal valve tissue (16.3 +/- 4.4 dB vs 9.8 +/- 3.3 dB, P < .0001). Backscatter values were greater (22.0 +/- 3.5 dB) in a group of 6 patients with aortic stenosis. Within the sclerosis group, the magnitude of backscatter was directly correlated (P < .05) with a subjective sclerosis score, and with transvalvular pressure gradient. Mean reproducibility was 2.4 +/- 1.8 dB (SD) between observers, and 2.3 +/- 1.7 dB (SD) between examinations. CONCLUSION: Measurement of backscatter from the valve leaflets of patients with ASC may be a feasible means of following the progression and treatment response of aortic sclerosis.     

Anisotropy of apparent backscatter in the short-axis view of mouse hearts

The goals of this investigation were to measure the anisotropy of backscattered ultrasound observed in the short-axis view of mouse hearts in systole and diastole and to compare these measurements with predictions from a computer simulation. Measurements of midmyocardial apparent backscatter were obtained from analyses of the hearts of seven wild-type mice using a clinical imaging system utilizing a linear array with a nominal center frequency of 13 MHz. A computer model simulating the short-axis view was implemented based on previous measurements of the angle of insonification dependence of myocardial backscatter and attenuation. Results demonstrate that the measured backscatter was largest for those myocardial regions corresponding to approximately perpendicular insonification relative to the myofibers and the smallest for regions of approximately parallel insonification, with the minimum to maximum values of apparent backscatter differing by approximately 10 dB. The measured anisotropy of backscatter was similar for end-systole and end-diastole and was in good agreement with the predicted anisotropy obtained from the computer simulations. (E-mail: mrh@wuphys.wustl.edu)     

High-frequency ultrasound m-mode imaging for identifying lesion and bubble activity during high-intensity focused ultrasound ablation

Effective real-time monitoring of high-intensity focused ultrasound (HIFU) ablation is important for application of HIFU technology in interventional electrophysiology. This study investigated rapid, high-frequency M-mode ultrasound imaging for monitoring spatiotemporal changes during HIFU application. HIFU (4.33 MHz, 1 kHz PRF, 50% duty cycle, 1 s, 2600?6100 W/cm²) was applied to ex vivo porcine cardiac tissue specimens with a confocally and perpendicularly aligned high-frequency imaging system (Visualsonics Vevo 770, 55 MHz center frequency). Radio-frequency (RF) data from M-mode imaging (1 kHz PRF, 2 s × 7 mm) was acquired before, during and after HIFU treatment (n = 12). Among several strategies, the temporal maximum integrated backscatter with a threshold of +12 dB change showed the best results for identifying final lesion width (receiver-operating characteristic curve area 0.91 ± 0.04, accuracy 85 ± 8%, compared with macroscopic images of lesions). A criterion based on a line-to-line decorrelation coefficient is proposed for identification of transient gas bodies.     

Developmental changes in integrated ultrasound backscatter from embryonic blood in vivo in mice at high US frequency

Mouse blood imaged using high-frequency ultrasound (US) is more echogenic in embryos than in adults. Studying changes in blood echogenicity in embryos may be of fundamental interest in studies on the genetic regulation of normal and abnormal blood development in mutant mice. Embryonic red blood cells (RBCs) are large and nucleated in midgestation but decrease in size and become enucleated as they mature. We therefore hypothesised that these structural alterations are responsible for variations in echogenicity of embryonic blood with gestational age and development. The objective of the current study was to quantify these structural changes in echogenicity (echo brightness) and apparent integrated backscatter (AIB) from embryonic blood at high US frequencies in vivo in mice. Results from anaesthetised pregnant mice studied using transcutaneous US showed that echogenicity of embryonic blood in the heart, aorta and umbilical cord and AIB within the heart chambers peaked at embryonic day (ED) 13.5 and then decreased progressively toward term. Between EDs 13.5 and 17.5 (near term), RBC mean cell volume decreased from 133 to 109 fL, haematocrit increased from 12 to 34%, and the percentage of nucleated RBCs decreased from 59 to 2%. Relative to younger ages, RBC nuclei at ED 13.5 were small and dense (pyknotic) which may have contributed to the peak in echogenicity and AIB at this age. To calculate the AIB, radiofrequency (RF) signals with centre frequencies of 28 MHz and 35 MHz were integrated over the 16- to 35-MHz and 21- to 42-MHz frequency range, respectively. At 28 MHz, mean apparent integrated backscatter of blood in the embryonic heart increased significantly from 0.0023 +/- 0.0004 Sr.cm(-1) (mean +/- SEM) at ED 12.5 to peak at 0.0037 +/- 0.0005 Sr.cm(-1) at ED 13.5. The mean AIB then decreased progressively with advancing gestation to 0.0002 +/- 0.0001 Sr.cm(-1) at ED 17.5. At 35 MHz, the mean AIB changed similarly with gestational age, except that values were lower than at 28 MHz at all ages. Higher attenuation of US at 35 MHz than at 28 MHz in tissue likely accounted for the lower AIB of blood insonified at 35 MHz. We speculate that developmental changes in red cell morphology are responsible for the observed changes in echogenicity and AIB of embryonic blood with gestational age in mice.     

Characterization of anisotropic myocardial backscatter using spectral slope, intercept and midband fit parameters

The specific myocardial structural components that contribute to the observed level of backscatter from the heart and its dependence on the angle of insonification have not been completely identified: The objectives of this study were to measure the anisotropy of backscatter from myocardium using the approach first introduced by Lizzi et al. [J Acoust Soc Am 73, 1366-1373 (1983)] and to use the extracted spectral parameters (spectral slope, intercept and midband fit) to characterize changes in the apparent scatterer size, spatial concentration and acoustic impedance properties as functions of the angle of insonification. Backscatter measurements were performed in vitro on eight cylindrical formalin-fixed lamb myocardial specimens using a 5 MHz focused transducer. The backscattered spectral data as a function of angle of insonification relative to the myocardial fiber direction were analyzed over the frequency range of 4 to 6 MHz. The spectral parameters describing features of backscatter were determined by applying a linear fit to attenuation-compensated normalized spectra. Results show that values of the spectral slope do not exhibit a significant dependence on the angle of insonification within uncertainties; however, the zero-frequency intercept showed clear anisotropy and was found to be a maximum for insonification perpendicular to the predominant myofiber orientation and a minimum for parallel insonification. A comparison of midband fit values at 5 MHz with attenuation-compensated integrated backscatter values showed excellent agreement for all angles of insonification. These data suggest that measurements of spectral slope, intercept, and midband fit can provide insights regarding aspects of tissue microstructure underlying the observed anisotropy of myocardial scattering properties. Measurements of the slope parameter suggest a very modest change in effective scatterer size with angle of insonification. However, the observed anisotropy in intercept and midband fit and apparent absence of anisotropy in the spectral slope suggests an angle of insonification dependence of acoustic concentration, the combination of effective spatial scatterer concentration and acoustic impedance properties, without a significant contribution from changes in effective scatterer size.     

Cyclic variation of ultrasound backscatter in normal myocardium is view dependent: clinical studies with a real-time backscatter imaging system

Real-time ultrasound backscatter imaging is a new method of evaluating relative integrated backscatter in a clinically applicable manner. The potential clinical utility of real-time backscatter imaging of diseased tissue depends on recognition of normal variations in cyclic backscatter when measured from different echocardiographic image orientations. The view dependence of cyclic backscatter variation was studied in normal human volunteers. In twenty normal male subjects (mean age 28 +/- 5 years) cyclic variation in integrated backscatter (diastolic minus systolic backscatter) was assessed in multiple left ventricular regions with four standard two-dimensional echocardiographic views (parasternal long-axis and short-axis views, and apical two-chamber and four-chamber views). M-mode backscatter imaging was performed from the standard parasternal long-axis view. Cyclic variation in backscatter was present in the septum only when imaged from the parasternal long-axis view (2.7 +/- 3.1 [standard deviation] decibels [dB], p less than 0.01 for diastole versus systole). The posterior wall of the left ventricle demonstrated cyclic variation of integrated backscatter when imaged from both the parasternal long-axis (4.6 +/- 1.6 dB, p less than 0.01) and short-axis views (2.8 +/- 2.2 dB, p less than 0.01). Cyclic variation in integrated backscatter was not demonstrated in inferoseptal, septal, or lateral wall regions when imaged from the parasternal short-axis view. The apical views did not demonstrate cyclic variation in integrated backscatter in any of the segments studied.(ABSTRACT TRUNCATED AT 250 WORDS)     

Measurements of ultrasonic attenuation properties of midgestational fetal pig hearts

The objectives of this study were to measure the relative attenuation properties of the left and right ventricles in fetal pig hearts and to compare the spatial variation in attenuation measurements with those observed in previously published backscatter measurements. Approximately 1.0-mm-thick, short-axis slices of excised, formalin-fixed heart were examined from 15 midgestational fetal pigs using a 50-MHz single-element transducer. Measurements of the attenuation properties demonstrate regional differences in the left and right ventricular myocardium that appear consistent with the previously reported regional differences in apparent integrated backscatter measurements of the same fetal pig hearts. For regions of perpendicular insonification relative to the myofiber orientation, the right ventricular free wall showed larger values for the slope of the attenuation coefficient from 30-60 MHz (1.48 +/- 0.22 dB/(cm x MHz) (mean +/- SD) and attenuation coefficient at 45 MHz (46.3 +/- 7.3 dB/cm [mean +/- SD]) than the left ventricular free wall (1.18 +/- 0.24 dB/(cm x MHz) and 37.0 +/- 7.9 dB/cm (mean +/- SD) for slope of attenuation coefficient and attenuation coefficient at 45 MHz, respectively). This attenuation study supports the hypothesis that intrinsic differences in the myocardium of the left and right ventricles exist in fetal pig hearts at midgestation.     

Diagnosis of recent myocardial infarction with quantitative backscatter imaging: preliminary studies

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)     

Age-related alterations of cardiac tissue microstructure and material properties in Fischer 344 rats

The cardiac aging process is accompanied by global mechanical dysfunction that reflects increased myocardial stiffness. Accordingly, age-related changes in microscopic material properties of myocardium were delineated with high-frequency ultrasound (US) (30 to 44 MHz) tissue characterization methods for aging Fischer 344 rats at 6 (adult), 18 (aged), and 24 (senescent) months of age. The excised lateral wall of the left ventricle of rats (n = 10 per group) was insonified with a 50-MHz acoustic microscope for determination of integrated backscatter, backscatter coefficient and attenuation coefficient. Histological and biochemical analyses for collagen content and cardiac myocyte diameter were performed. Collagen concentration increased progressively with age, with the greatest increments occurring from 6 to 18 months (38.0 +/- 6.3 to 53.0 +/- 7.1 mg/g dry wt), and leveling off at 24 months (60.0 +/- 7.4 mg/g dry wt). Tissue microscopic material properties also changed progressively from 6 to 24 months of age, as determined by US methods: integrated backscatter increased (-44.7 +/- 1.8 vs. -40.8 +/- 1.9 dB, p < 0.05), attenuation increased (47.1 +/- 5.9 to 65.3 +/- 7.8 dB/cm, p < 0.05), and the backscatter coefficient increased (0.73 +/- 0.16 x 10(-5) to 3.76 +/- 1.6 x 10(-5) cm(-1), p < 0.05), from 6 to 24 months of age in each case. Age-related alterations in indices of cardiac microscopic material properties were closely correlated with the changes in cardiac microstructure. Ultrasonic tissue characterization may prove to be a sensitive tool to monitor changes in the cardiac microstructure, such as increased collagen deposition, that occur within age-related diastolic dysfunction.     

Left ventricular remodeling, mechanics, and tissue characterization in congenital aortic stenosis.

BACKGROUND: As the response of the myocardium to pressure overload is age-dependent, this study was designed to examine left ventricular (LV) remodeling, mechanics, and tissue characterization in children with moderate congenital aortic stenosis. METHODS: We studied by echocardiography Doppler 22 patients (mean age 12.4 +/- 5.6 years) with peak and mean transvalvular gradient of 63 +/- 6 and 32 +/- 4 mm Hg, respectively. In addition, 30 age- and body surface area-matched participants with structurally normal hearts were used as a control group. Sex- and age-specific cut-off levels for LV mass/height(2.7) and relative wall thickness were defined to assess LV geometry. As a load-independent index of myocardial contractility, the relation between the rate-corrected velocity of circumferential fiber shortening both at endocardium and midwall, and meridional end-systolic stress was assessed. In addition, LV diastolic function was also evaluated by the mitral flow indexes. Finally, ultrasonic tissue characterization of the LV myocardium was performed by calculating the magnitude of cyclic variation, which reflects the intramural contractile function, and the averaged myocardial intensity of integrated backscatter, which is directly related to the myocardium collagen content. RESULTS: The endocardial velocity of circumferential fiber shortening endocardium and meridional end-systolic stress relationship was within the normal range (mean +/- 2SD) in 18 of 22 patients (81.8%), and midwall velocity of circumferential fiber shortening at endocardium and meridional end-systolic stress was normal in all 22 patients. No mitral flow index of LV diastolic function was significantly different between aortic stenosis group and normal participants. In our study population, 16 of 22 patients (72.7%) showed normal LV geometry, 3 (13.6%) had a pattern of concentric remodeling, and 3 (13.6%) concentric hypertrophy. LV hypertrophy was not marked (left ventricular mass index [LVMI] < 51 g/m(2.7)) in any patient. Finally, compared with control participants our study population showed, both at interventricular septum and posterior wall, comparable values of cyclic variation integrated backscatter, but significantly higher values of averaged myocardial integrated backscatter intensity (P <.01). CONCLUSIONS: In children with moderate congenital aortic stenosis, the total amount of myocardial collagen was increased despite normal LV myocardial contractility and diastolic function. Furthermore, LV remodeling was abnormal in only about a quarter of our patients and none had more than mild hypertrophy. Although the majority of these patients do not have markers now recognized to predict higher risk of cardiovascular events, the long-term significance of myocardial fibrosis and its response to treatment remain to be investigated.     

Enhanced method for predicting left ventricular reverse remodeling after surgical repair of aortic regurgitation: application of ultrasonic tissue characterization.

To predict left ventricular (LV) reverse remodeling after surgical repair of aortic regurgitation, we examined 30 patients with aortic regurgitation accompanying LV dilatation by myocardial tissue characterization with integrated backscatter method. Before and after operation, the magnitude of cyclic variation of integrated backscatter (CVIB) was obtained from anterior septum and posterior wall, and averaged value was calculated in each patient. Before operation, LV end-diastolic dimension, fractional shortening, and LV end-diastolic pressure were not significantly different between the patients with (group GR) and without (group PR) decreased LV end-diastolic dimension after operation. Under these conditions, CVIB, which was 9.6 +/- 1.0 dB from healthy volunteers, was significantly greater in group GR, 5.7 +/- 1.4 dB, than that in group PR, 3.8 +/- 0.8 dB (P =.0003). The patients with CVIB >/= 4 before operation were expected to have reverse remodeling after operation with a sensitivity of 79%, a specificity of 82%. These data indicate that preoperative CVIB from the left ventricle provides pivotal information for predicting reverse remodeling after operation for aortic regurgitation in addition to the conventional echocardiographic parameters.     

In vitro strain measurement in the porcine antrum using ultrasound doppler strain rate imaging

Strain rate imaging (SRI) enables study of deformation in soft tissues. The aim of this study was to evaluate the accuracy of SRI in measuring strain in the porcine antral wall in vitro. An experimental set-up enabled controlled distension of a porcine stomach in a saline reservoir. Radial strain obtained by SRI was compared with radial strain calculated from B-mode ultrasonography. Circumferential strain obtained by SRI was compared with circumferential strain calculated from sonomicrometry. The agreement between radial strain values measured by SRI and B-mode, along and across several ultrasound (US) beams, using US frequency 6.7 MHz and strain length (SL) = 1.9 mm was = -1.0 +/- 12.1% and 0.5 +/- 13.4%, respectively (mean difference +/- 2SD%) and it was better than with SL 1.2 mm. Compared with sonomicrometry, SRI-determined circumferential strain using 6.7 MHz and SL = 1.9 mm was less accurate, whether averaging along or across several US beams (-9.2 +/- 46.7% and 13.8 +/- 51.2%, respectively). In conclusion, SRI gave accurate measurement of radial strain of the antral wall, but seemed to be less accurate for measurement of circumferential strain for this in vitro set-up.     

Regional variation in the measured apparent ultrasonic backscatter of mid-gestational fetal pig hearts

The goal of this study was to characterize and compare regional backscatter properties of fetal hearts through measurements of the apparent integrated backscatter. Sixteen excised, formalin-fixed fetal pig hearts, representing an estimated 53 to 63 days of gestation, were investigated. Spatially localized measurements of integrated backscatter from these specimens were acquired using a 50 MHz single-element transducer. The apparent integrated backscatter measurements demonstrate different patterns of backscatter from the myocardium of the right ventricle compared with that of the left ventricle. These backscatter measurements appear to be consistent with the anisotropy of the fiber orientation observed in histologic assessment of the same specimens. For each of the 16 hearts, the apparent integrated backscatter from the right ventricular myocardium was larger than that from the left ventricular myocardium, exhibiting mean apparent backscatter values of –35.9 ± 2.0 dB and –40.1 ± 1.9 dB (mean ± standard deviation; n = 16; p < 0.001), respectively. This study suggests that the intrinsic ultrasonic properties of the left and right ventricular myocardium are distinct in fetal pig hearts at mid-gestation. (E-mail: mrh@wuphys.wustl.edu)     

Ultrasonic backscatter and attenuation (11-27 MHz) variation with collagen fiber distribution in ex vivo human dermis

This ex vivo study explores the relationship of ultrasonic attenuation and backscatter to dermal microarchitecture by comparing ultrasonic measurements of these parameters (11-27 MHz) to a microscopic analysis of three parameters describing the collagen distribution (mean thickness and spacing of collagen bundles along the insonification direction and the percent area occupied by collagen). Skin samples (N= 31) were obtained from patients undergoing breast or abdominal reduction surgery. Radio-frequency (rf) signals were acquired in a B-scan format using an ultrasound system developed for skin imaging (Ultrasons Technologies, Tours, France). Ultrasonic data were analyzed to calculate average integrated backscatter (IBS in dB) and frequency dependence of backscatter (n, dimensionless) of each specimen at depths centered approximately 370,620 and 880 microm beneath the skin surface. Average integrated attenuation coefficient (IA in dB.cm(-1)) and frequency dependence of attenuation coefficient (beta in dB.cm(-1).MHz(-1)) were estimated across the depth between 240 and 1,000 microm. The three collagen distribution parameters were estimated using digitized microcopic fields from matched regions of histological sections stained with hematoxylin-eosin-saffron. No significant correlation was identified between collagen distribution parameters and IA or beta. For the most superficial depth studied in abdominal skin, n was inversely correlated to collagen bundle thickness (r = -0.67,p = 0.002) and percent area (r = -0.65,p = 0.003). At the same depth, IBS was inversely correlated to percent area of collagen (r = -0.51,p = 0.03). The rather high collagen packing (48 to 82% area) measured in histological sections and the inverse relationship observed between IBS and percent area of collagen suggest that a packing factor should be included in models relating skin collagen distribution to ultrasound spectral parameters. A better understanding of the relationship between ultrasound parameters and the microarchitecture of the dermis should help to interpret changes in ultrasonic parameters observed during in vivo ultrasonic skin examinations.     

Cardiac valve bioprosthesis frames and donor aortic valve commissural spacing

Intercommissural distances in 300 porcine and 200 human aortic valves were expressed as percentages of the smallest dimension. The right coronary cusp was generally the largest but whereas in porcine valves the noncoronary cusp was generally the smallest, in human valves this status was shared about equally between the noncoronary and left coronary cusps.The dominant pattern was with all three dimensions unequal (difference > 5%). Seventy-seven percent of the porcine valves and 84% of the human valves were in this category. The asymmetry existed in both clockwise and anticlockwise directions.Accurate concordance between the valve commissures and supporting frame struts is essential in the fabrication of bioprosthetic cardiac valves and the optimum utilisation of donor aortic valves would result from using frames in which the strut angles were 110°, 120°, 130° for porcine valves and 111°, 120°, 129° for human valves, in both clockwise and anticlockwise patterns.     

Estimating myocardial attenuation from M-mode ultrasonic backscatter

The objective of this investigation was to determine whether measurements of myocardial attenuation can be obtained from analyses of M-mode images. We exploited the inherent anisotropy of myocardial properties as a means of systematically varying the attenuation to evaluate this M-mode image-based method for myocardial tissue characterization. A commercially available ultrasonic imaging system was used to acquire M-mode images of 24 excised cylindrical specimens from six formalin-fixed sheep hearts that were analyzed using video signal analysis. Data were compensated for the presence of bright intramural myocardial echoes, a potentially significant contributor to uncertainty in measurements of attenuation from backscattered ultrasound. The estimated attenuation coefficient in dB/cm at an effective center frequency of 2.75 MHz as a function of angle of insonification for measurements obtained from analyses of M-mode images is presented. Given a linear frequency-dependence of attenuation in myocardial tissue over frequencies ranging from 1.5 MHz to 8 MHz, as has been previously reported, M-mode image-based analyses were used to estimate the slope of attenuation. Results showed slopes of attenuation (over a -10 dB transmit bandwidth of 1.875 MHz to 3.75 MHz) ranging from 1.00 +/- 0.07 to 1.81 +/- 0.08 dB/(cm.MHz) for perpendicular and parallel insonification, respectively. These values were in good agreement with contemporaneously measured values (0.99 +/- 0.02 to 1.77 +/- 0.04 dB/(cm.MHz)) obtained over a frequency bandwidth of 4 MHz to 7 MHz using a through-transmission radio-frequency-based approach. These data suggest that robust measurements of myocardial attenuation can be obtained from analyses of M-mode images and that this method may be diagnostically feasible in the clinical setting.     

Detection of blood coagulation and clot formation using quantitative ultrasonic parameters

The ultrasonic parameters of sound velocity, attenuation and integrated backscatter were applied to detect the process of coagulation and clot formation in porcine blood. Fresh porcine blood containing 15% anticoagulant solution was collected. Blood samples with a hematocrit of 45% were obtained by reconstituting the packed erythrocytes with the separated plasma for ultrasound measurements performed with a 10-MHz focused transducer. A 24-mL aliquot of blood was placed in a container and 12 mL of 0.2 mol/L CaCl2 solution was added to induce clot formation. In each measurement, radio-frequency signals of the blood digitized at 100 MHz were collected for 50 min at a temporal resolution of 1 A-line per s. Results showed that all of the parameters increased within the initial 3 min and, then, immediately decreased dramatically as the CaCl2 solution was added. Subsequently, the sound velocity gradually increased with time and the integrated backscatter and attenuation increased in accordance with blood coagulation until approximately 500 and 2600 s, respectively. The integrated backscatter, attenuation and sound velocity can be divided into different stages, including red cell aggregation, reduction in hematocrit, blood coagulation and clot formation, corresponding to variations in the physical and chemical properties of the blood. The integrated backscatter, attenuation and sound velocity increased because of the changes in blood properties during the process of coagulation and clot formation: by 8.2 dB, 0.65 dB/cm, and 0.6%, respectively. These results provide a feasibility for further applying ultrasonic parameters to in vivo monitor the progress of clotting and thrombosis research.     

Molecular imaging of vascular endothelial growth factor receptor 2 expression using targeted contrast-enhanced high-frequency ultrasonography.

OBJECTIVE: The aim of our study was to investigate the use of targeted contrast-enhanced high-frequency ultrasonography for molecular imaging of vascular endothelial growth factor receptor 2 (VEGFR2) expression on tumor vascular endothelium in murine models of breast cancer. METHODS: Highly invasive metastatic (4T1) and nonmetatstatic (67NR) breast cancer cells were implanted in athymic nude mice. Tumors were examined in vivo with targeted contrast-enhanced high-frequency ultrasonography using a scanner with a 40-MHz probe. Randomized boluses of ultrasound contrast agents (UCAs) conjugated with an anti-VEGFR2 monoclonal antibody or an isotype control antibody (immunoglobulin G) were injected into the animals. Sonograms were analyzed by calculation of the normalized video intensity amplitudes caused by backscatter of the bound UCA. After ultrasonography, the tumor samples were harvested for analysis of VEGFR2 expression by immunoblotting and immunocytochemistry. RESULTS: The mean video intensity amplitude caused by backscatter of the retained VEGFR2-targeted UCA was significantly higher than that of the control UCA (mean +/- SD: 4T1 tumors, 15 +/- 3.5 versus 7 +/- 1.6 dB; P < .01; 67NR tumors, 50 +/- 12.3 versus 12 +/- 2.6 dB; P < .01). There was a significant difference in VEGFR2-targeted UCA retention between 4T1 and 67NR tumors (normalized video intensity amplitudes, 15 +/- 3.5 and 50 +/- 12.3 dB, respectively; P < .001), and this correlated well with relative VEGFR2 expression in the two tumor types. CONCLUSIONS: Targeted contrast-enhanced high-frequency ultrasonography may enable in vivo molecular imaging of VEGFR2 expression on the tumor vascular endothelium and may be used for noninvasive longitudinal evaluation of tumor angiogenesis in preclinical studies.