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Optimization of acoustic scattering from dual-frequency driven microbubbles at the difference frequency.

The second harmonic radiation of acoustically driven bubbles is a useful discriminant for their presence in clinical ultrasound applications. It is useful because the scatter from a bubble at a frequency different from the driving can have a contrast-to-tissue ratio better than at the drive frequency. In this work a technique is developed to optimize the scattering from a microbubble at a frequency different from the driving. This is accomplished by adjusting the relative phase and amplitudes of the components of a dual-frequency incident ultrasound wave form. The investigation is focused primarily on the example of dual-mode driving at frequencies of 1 MHz and 3 MHz, with the scattering optimized at 2 MHz. Bubble radii of primary interest are 0.5 to 2 microm and driving amplitudes to 0.5 atm. Bubbles in this size range are sensitive to modulation of driving. It is shown that an optimal forcing scheme can increase the target response eightfold or more. This suggests new applications in imaging and in bubble detection.

Animals↗

Acoustic pressure amplitude thresholds for rectified diffusion in gaseous microbubbles in biological tissue.

One of the mechanisms often suggested for the biological action of ultrasonic beams irradiating human tissues is concerned with the presence in the tissues of minute gaseous bubbles which may, under the influence of the ultrasonic field be stimulated to grow to a size a which resonance or collapse occurs with severe associated shear stresses. The evidence for the existence of microbubbles in tissues is reviewed. The results of calculations, using two existing theoretical models, of the peak pressure threshold as a function of frequency are presented. The frequency is normalized with the resonant frequency of the bubble, and results are presented for three bubble radii (1, 2, and 3.5 micrometer) and for different values of the gas concentration in the tissue between 0.1 and 1. The results from two models differ suggesting that an improved model and better experimental data for the threshold calculations would be appropriate for further calculations. The thresholds calculated range below the peak pressure amplitudes used in continuous wave diagnostic instruments, indicating the need for a more careful investigation both of this damage mechanism and of the exposures used in routine diagnosis. The results of calculations for typical (transient) exposure conditions from pulse-echo equipment are presented, indicating that rectified diffusion and stable cavitation are improbable phenomena in these circumstances.

Animals↗

Experimental frequency spectra of ultrasound transmission through two-dimensional ensembles of trapped microbubbles.

The frequency spectra of transmission coefficients for ultrasound passing through a sheet of gas-filled micropores have been measured using incident waves with amplitudes up to 2.4 x 10(4) Pa. It is found that as the amplitude of the incident wave is increased, the peak frequency of transmission loss through the two-dimensional trapped-bubble ensemble shifts to lower values by as much as 47%. The experiments indicate that the shifting is caused by the net displacements of air-water-membrane triple-phase lines and the air loss of the trapped bubble, which are due to the radiation force and microbubble production produced by the incident wave. Results of the experiments and possible theoretical explanations are discussed.

Ultrasonics↗

Sound scattering and localized heat deposition of pulse-driven microbubbles

The sound scattering of free microbubbles released from strongly driven ultrasound contrast agents with brittle shell (e.g., Sonovist) is studied numerically. At high peak pressure of the driving pulses, the bubbles respond nonlinearly with cross sections pronouncedly larger than in the linear case; a large portion of the energy is radiated into high frequency ultrasound. Subsequent absorption of these high frequencies in the surrounding liquid (blood) diminishes the effective scattering cross section drastically. The absorption results in highly localized heating, with a substantial temperature rise within the first few microm from the bubble surface. The maximum heating in 1 microm distance is strongly dependent on driving pressure. Temperature elevations of more than 100 K can be achieved for amplitudes of Pa approximately 30 atm, which coincides with the highest pressures used in ultrasound diagnostics. The perfectly spherical collapses assumed here occur rarely, and the heating is highly localized and transient (approximately 10 micros). Therefore, a thermal hazard would only be expected at driving pressures beyond the diagnostic range.

Journal Article↗

Bacterial stress responses to 1-megahertz pulsed ultrasound in the presence of microbubbles.

Members of a panel of stress-responsive biosensors have been used to study the effect of megahertz frequency ultrasound on Escherichia coli. Insonification causes acoustic cavitation, the collapse of oscillating microbubbles in solution, which can damage bacterial cells. A focused 1-MHz ultrasound transducer, capable of generating a spatial peak pulse average intensity of 500 W/cm2, was used to treat liquid bacterial cultures. Stress-responsive promoters fused to luxCDABE allowed the continuous measurement of light produced as a result of protein damage, DNA damage, oxidative stress, and membrane perturbation. A promoter responsive to ammonia limitation was not transcriptionally activated under test conditions. In contrast to bacteria in exponentially growing cultures, those in stationary-phase cultures were more resistant to the effects of ultrasound treatment. Quantification of the degree of acoustic cavitation due to symmetric bubble collapse was measured by a 20-MHz passive transducer, the output of which appears to be only partially correlated with cellular damage and survival. The methods and results summarized here provide the basis for further investigation into applications, including the purification of water samples.

Ammonia↗

Interaction with leukocytes: phospholipid-stabilized versus albumin-shell microbubbles.

PURPOSE: To confirm that BR14 microbubbles (MBs) can be phagocytosed by activated leukocytes, to determine their stability after phagocytosis, and to evaluate how such characteristics influence the fate of neutrophils containing MBs after insonation. MATERIALS AND METHODS: BR14 and human albumin MBs (2 x 10(7)/mL) were incubated with activated human neutrophils (2 x 10(6)/mL) to allow phagocytosis. Deflation rate of the phagocytosed MBs after pulsed insonation (one burst per second for 5 seconds) at 1.8 MHz with peak negative pressure of -540 kPa or -1,340 kPa, lactate dehydrogenase (LDH) leakage, and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling stain-positive cell count after insonation were compared between the two agents. RESULTS: At -540 kPa, phagocytosed MBs remained nearly unchanged for both agents after insonation. At -1,340 kPa, although human albumin MBs were disrupted on the first or second burst, BR14 MBs remained undisrupted. After -540-kPa insonation, a similar number of apoptotic cells appeared in neutrophils containing human albumin and BR14 MBs. At -540 kPa, LDH leakage was limited in human albumin MBs and BR14 MBs. At -1,340 kPa, LDH leakage was significantly increased in human albumin MBs and BR14 MBs (P <.01, both vs -540 kPa). Apoptotic cells were significantly decreased in human albumin MBs and BR14 MBs (P <.01, both vs -540 kPa). LDH leakage was lower and apoptotic cell count was greater in BR14 MB-containing neutrophils than in human albumin MB-containing neutrophils (both P <.01). CONCLUSION: Compared with human albumin MBs, BR14 MBs were more stable after phagocytosis with insonation. This stability is associated with less disruption and greater induction of apoptosis in leukocytes after relatively high-pressure insonation in the range for diagnostic use.

Adult↗

Ultrasonic contrast enhancement of tissue by encapsulated microbubbles.

Pilot studies have shown that gelatin encapsulated microbubbles (GEM) are readily demonstrable ultrasonic contrast agents. GEM injected into hepatic and renal blood vessels resulted in increased echogenicity of the parenchyma in normal rabbits. VS carcinoma implanted in the rabbit liver and kidney showed significantly different echogenicity from that of normal parenchyma following GEM injection. Echogenic bubbles were delivered to different regions by varying the size of GEM.

Animals↗

Hepatic tumors: US contrast enhancement with CO2 microbubbles.

Enhancement of hepatic tumors on sonograms by injection of carbon dioxide microbubbles into the hepatic artery as a contrast material (enhanced ultrasonography) was performed in 43 patients with various histologically confirmed hepatic tumors. Enhanced sonograms were classified into five patterns according to the relative changes of the echo levels between the tumor and the nontumorous parenchyma of the liver as a result of enhancement: hyperechoic change, isoechoic change, hypoechoic change with hyperechoic rim (rim sign), marginal spotty hyperechoic change, and internal spotty hyperechoic change. Eighty-eight percent of hepatocellular carcinomas showed hyperechoic change, 70% of metastatic tumors exhibited hypoechoic change with the rim sign. The marginal spotty hyperechoic change and the internal spotty hyperechoic change were specific for cavernous hemangioma and fibrous granuloma, respectively. This method of enhancement is useful in assessing the nature of liver tumors and in the detection of small nodules in the liver.

Carbon Dioxide↗

Hepatic focal nodular hyperplasia: specific findings at dynamic contrast-enhanced US with carbon dioxide microbubbles.

Dynamic contrast material-enhanced ultrasonography (US) with intraarterial infusion of carbon dioxide microbubbles was performed for four cases of histologically proved focal nodular hyperplasia (FNH) in four patients and for 167 cases of various hepatic nodules in 144 patients. No complications due to dynamic US were observed in any of the 148 patients. All FNH nodules were less than 3 cm in diameter. Consistent specific findings of FNH were not obtained with US, computed tomography, magnetic resonance imaging, radiocolloid scanning, or angiography in the four cases of FNH. In contrast, the characteristic vascular pattern (ie, early central hypervascular supply with centrifugal filling to the periphery at the arterial phase and a uniform or lobulated dense stain at the capillary phase) was observed in all four cases of FNH with dynamic US. This vascular pattern demonstrated in FNH with dynamic US was not seen in any of the 167 hepatic nodules, including 44 small hepatocellular carcinomas less than 3 cm in diameter. Therefore, the newly developed, dynamic contrast-enhanced US technique seems to be extremely sensitive and specific for diagnosing FNH and is useful in the differentiation of FNH from other hepatic tumors, especially hepatocellular carcinoma.

Adult↗

Small hepatocellular carcinoma: diagnosis with US angiography with intraarterial CO2 microbubbles.

Ultrasonographic (US) angiography enhanced with intraarterial CO2 microbubbles, a contrast material used in US imaging, was performed of 103 histologically proved hepatocellular carcinomas (HCCs) smaller than 3 cm in diameter in 95 patients. The detection rate for hypervascular HCC with US angiography was compared with the rate of detection with conventional angiography, digital subtraction angiography (DSA), and computed tomography (CT) after intraarterial injection of iodized oil. Sensitivity in detection of hypervascular HCCs with US angiography was 86% (89 of 103 HCCs), compared with 63% (44 of 70 HCCs) detected with conventional angiography, 70% (23 of 33 HCCs) with DSA, and 82% (75 of 91 HCCs) with CT with iodized oil. US angiography depicted small hypervascular HCCs, especially those less than 1 cm in diameter, and helped clarify vascularity as isovascular or hypovascular in angiographically undetectable HCCs. Findings at US angiography assisted the choice of a therapeutic strategy for treatment of HCC, such as transarterial therapy, percutaneous ethanol injection therapy, or resection.

Angiography, Digital Subtraction↗

Hepatocellular carcinoma in adenomatous hyperplasia: detection with contrast-enhanced US with carbon dioxide microbubbles.

Adenomatous hyperplasia (AH) of the liver is considered a precancerous lesion because hepatocellular carcinoma (HCC) has been found in nodules of AH at histologic examination. Contrast material-enhanced ultrasound (US) with intraarterial infusion of carbon dioxide microbubbles was performed in four patients with HCC in AH lesions. In all four patients, a "hyperechoic focus in an area of hypoechoic change" in relation to the adjacent liver was demonstrated, compatible with the minimal vascular change associated with neoplastic transformation and carcinoma development. Because early detection of the minute cancer foci that may develop in an AH nodule is clinically important, this contrast-enhanced US technique may be a useful tool for the detection of these associated lesions.

Adenoma↗

Analysis of regional cerebral blood flow in dogs, with an experimental microbubble-based US contrast agent.

PURPOSE: To evaluate a perfluorochemical stabilized microbubble-based ultrasound (US) contrast agent (Imagent US, formulation AF0145) create regional cerebral blood flow (CBF) maps. MATERIALS AND METHODS: Intravenous injections of the contrast agent (0.16-0.20 mL/kg) were performed in four anesthetized dogs (9-14 kg). Serial coronal color-Doppler-amplitude and gray-scale US scans were obtained through a cranial window during injection of contrast material. Mean pixel intensity was analyzed on digitized images. Regional CBF was varied by modifying arterial partial pressure of carbon dioxide (PaCO2). Color-Doppler-amplitude and gray-scale mean pixel intensity and percentage change in mean pixel intensity after injection of contrast material were analyzed, by means of a bolus injection curve that plotted color mean pixel intensity per unit area versus time. RESULTS: Contrast material injection resulted in a statistically significant difference in color-Doppler-amplitude and gray-scale enhancement of brain. Color mean pixel intensity increased from 1.4 units +/- 1 (standard error) to 122 units +/- 6 in thalamus and from 9.7 units +/- 4.0 to 107 units +/- 9 in cortex (P < .0001). Gray-scale mean pixel intensity increased from 0.55 units +/- 0.20 to 33 units +/- 11 in thalamus and from 0.42 units +/- .06 to 10.5 units +/- 2.0 in cortex (P < .005). Correlation between the area under the color-Doppler-amplitude mean-pixelintensity curve and regional CBF in thalamus and cortex was significant (r = .815, P < .0001). CONCLUSION: Color-Doppler-amplitude US can be used to create regional CBF maps and shows potential for noninvasive evaluation of CBF in the critically ill newborn.

Animals↗

Twin fetuses: intravascular microbubble US contrast agent administration--early experience.

PURPOSE: To explore the feasibility of administering SH U 508A by using a single-needle procedure at ultrasonography (US) in twin pregnancies to confirm interfetal transfusion in monochorionic twins and delineate placental angioarchitecture in pregnancies with twin-twin transfusion syndrome. MATERIALS AND METHODS: Fourteen twin pregnancies were studied over 12 months: seven with monochorionic twins, including six with twin-twin transfusion syndrome; two of unknown chorionicity; and five with known dichorionic twins discordant for fetal karyotype or anomaly and undergoing selective feticide in the third trimester. Bolus injection of 100 microL/kg of estimated fetoplacental weight of 400 mg/mL of SH U 508A was performed in the intrahepatic vein of one twin, and evidence of interfetal transfusion was sought by means of digital analysis of power Doppler signals in the contralateral twin. RESULTS: Contralateral twin echo enhancement was seen in four of the nine ultimately histopathologically proved monochorionic twins. As expected, no evidence of echo enhancement in the contralateral twin was seen in any of the five dichorionic twin pregnancies. There was no evidence of fetal compromise associated with the procedure. CONCLUSION: These pilot results suggest that microbubbles can be used to demonstrate interfetal transfusion but not to delineate placental vascular anatomy.

Adult↗

Liver lesions: intermittent second-harmonic gray-scale US can increase conspicuity with microbubble contrast material-early experience.

The authors investigated the effect of intermittent second-harmonic gray-scale (ISHGS) ultrasonography (US) with SH U 508A microbubbles on the conspicuity of focal liver lesions. Twenty-three patients were included in the study. Images were analyzed subjectively and quantitatively. Objective lesion conspicuity was increased. In 12 of the 15 patients with liver malignancy, gray-scale defects were seen in previously unsuspected areas. ISHGS US may improve the sensitivity of US for liver lesions.

Adult↗

Nonpalpable breast lesions: evaluation with power Doppler US and a microbubble contrast agent-initial experience.

PURPOSE: To evaluate power Doppler ultrasonography (US) performed with a microbubble US contrast agent in the differentiation of nonpalpable breast lesions. MATERIALS AND METHODS: Fifty nonpalpable breast lesions in 50 patients were prospectively evaluated with power Doppler US before and after injection of the contrast agent SH U 508A. Lesion vascularity and the morphology of vessels on US scans were analyzed and were correlated with histologic results. RESULTS: Surgical excision revealed 22 cancers and 28 benign lesions. At nonenhanced power Doppler US, eight (36%) of 22 cancers and four (14%) of 28 benign lesions were vascular. At contrast agent-enhanced power Doppler US, 21 (95%) cancers and six (21%) benign lesions were vascular (P <.001). Irregular vessels were seen in three cancers and one benign lesion at nonenhanced power Doppler US and in 11 cancers and one benign lesion at contrast-enhanced power Doppler US. By using the presence of vascularity in the mass as the diagnostic criterion for malignancy, the sensitivity, specificity, and positive and negative predictive values of power Doppler US changed from 36%, 86%, 67%, and 63%, respectively, to 95%, 79%, 78%, and 96% after contrast agent injection. CONCLUSION: Contrast-enhanced power Doppler US was superior to nonenhanced power Doppler US in the demonstration and characterization of tumor vascularity in nonpalpable breast lesions. Contrast-enhanced power Doppler US may be useful for the differentiation between nonpalpable breast cancers and benign tumors.

Adult↗

Microbubble elimination during priming improves biocompatibility of membrane oxygenators.

We tested the hypothesis that platelet loss following blood contact with foreign materials is partly related to the presence of microbubbles of gas (gas nuclei) trapped in surface defects on the membrane material. Extracorporeal membrane oxygenator perfusions were conducted in two groups of sheep, with use of standard priming techniques for the oxygenator in one group and a new vacuum priming technique in the other group. The vacuum priming technique was developed to eliminate gas nuclei from the oxygenator surface. With denucleation priming, platelet loss during perfusion was markedly reduced, as was thrombus formation on the membrane surface. The platelet particle-size distribution curve showed a shift consistent with platelet aggregation with the standard priming technique but not with the vacuum priming technique. We conclude that the elimination of trapped gas nuclei from the membrane surface during priming reduces the initial interaction between blood elements and the foreign surface.

Animals↗

Detection of hepatocellular carcinoma by intraarterially enhanced ultrasonography with CO2 microbubbles. Comparison with DSA, dynamic CT, and Lipiodol CT.

PURPOSE: To evaluate the diagnostic value of contrast-enhanced US with CO2 microbubbles (EU) for HCCs. Detectability was compared with DSA, dynamic CT, and Lipiodol CT. MATERIAL AND METHODS: Forty-seven histologically proven HCCs detected with US were evaluated with EU, DSA, and dynamic CT. In 23 patients (35 lesions), Lipiodol CT was also performed. The size of the tumors ranged from 8 to 71 mm (average 28.1 mm); 24 lesions were smaller than 20 mm and 23 lesions were larger than 20 mm. RESULTS: Overall detection was possible in 40 of 47 lesions (85%) by EU, in 32 of 47 (68%) by DSA, in 33 of 47 (74%) by dynamic CT, and in 27 of 35 (77%) by Lipiodol CT. In tumors smaller than 20 mm, detection was possible in 21 of 24 lesions (88%) by EU, 14 of 24 (58%) by DSA, 14 of 24 (58%) by dynamic CT, and 11 of 17 (65%) by Lipiodol CT. CONCLUSION: EU has significant diagnostic value for detection of HCCs, particularly tumors smaller than 20 mm.

Angiography, Digital Subtraction↗

Eliminating high-order scattering effects in optical microbubble sizing.

Measurements of bubble size and velocity in multiphase flows are important in much research and many industrial applications. It has been found that high-order refractions have great impact on microbubble sizing by use of phase-Doppler anemometry (PDA). The problem has been investigated, and a model of phase-size correlation, which also takes high-order refractions into consideration, is introduced to improve the accuracy of bubble sizing. Hence the model relaxes the assumption of a single-scattering mechanism in a conventional PDA system. The results of simulation based on this new model are compared with those based on a single-scattering-mechanism approach or a first-order approach. An optimization method for accurately sizing air bubbles in water has been suggested.

Journal Article↗