Contrast-Enhanced Ultrasound to Assess Carotid Intraplaque Neovascularization.

Contrast-enhanced ultrasound (CEUS) is increasingly being used to identify patients with carotid plaques that are vulnerable to rupture, so-called vulnerable atherosclerotic plaques, by assessment of intraplaque neovascularization. A complete overview of the strengths and limitations of carotid CEUS is currently not available. The aim of this systematic review was to provide a complete overview of existing publications on the role of CEUS in assessment of carotid intraplaque neovascularization. The systematic review of the literature yielded 52 studies including a total of 4660 patients (mean age: 66 y, 71% male) who underwent CEUS for the assessment of intraplaque neovascularization. The majority of the patients (76%) were asymptomatic and had no history of transient ischemic attack (TIA) or stroke. The assessment of intraplaque neovascularization was mostly performed using a visual scoring system; several studies used time-intensity curves or dedicated quantification software to optimize analysis. In 17 studies CEUS was performed in patients before carotid surgery (endarterectomy), allowing a comparison of pre-operative CEUS findings with histologic analysis of the tissue sample that is removed from the carotid artery. In a total of 576 patients, the CEUS findings were compared with histopathological analysis of the plaque after surgery. In 16 of the 17 studies, contrast enhancement was found to correlate with the presence and degree of intraplaque neovascularization on histology. Plaques with a larger amount of contrast enhancement had significantly increased density of microvessels in the corresponding region on histology. In conclusion, CEUS is a readily available imaging modality for the assessment of patients with carotid atherosclerosis, providing information on atherosclerotic plaques, such as ulceration and intraplaque neovascularization, which may be clinically relevant. The ultimate clinical goal is the early identification of carotid atherosclerosis to start early preventive therapy and prevent clinical complications such as TIA and stroke.

SUSHI: Sparsity-Based Ultrasound Super-Resolution Hemodynamic Imaging.

Identifying and visualizing vasculature within organs and tumors has major implications in managing cardiovascular diseases and cancer. Contrast-enhanced ultrasound scans detect slow-flowing blood, facilitating noninvasive perfusion measurements. However, their limited spatial resolution prevents the depiction of microvascular structures. Recently, super-localization ultrasonography techniques have surpassed this limit. However, they require long acquisition times of several minutes, preventing the detection of hemodynamic changes. We present a fast super-resolution method that exploits sparsity in the underlying vasculature and statistical independence within the measured signals. Similar to super-localization techniques, this approach improves the spatial resolution by up to an order of magnitude compared to standard scans. Unlike super-localization methods, it requires acquisition times of only tens of milliseconds. We demonstrate a temporal resolution of ~25 Hz, which may enable functional super-resolution imaging deep within the tissue, surpassing the temporal resolution limitations of current super-resolution methods, e.g., in neural imaging. The subsecond acquisitions make our approach robust to motion artifacts, simplifying in vivo use of super-resolution ultrasound.

Fast Vascular Ultrasound Imaging With Enhanced Spatial Resolution and Background Rejection.

Ultrasound super-localization microscopy techniques presented in the last few years enable non-invasive imaging of vascular structures at the capillary level by tracking the flow of ultrasound contrast agents (gas microbubbles). However, these techniques are currently limited by low temporal resolution and long acquisition times. Super-resolution optical fluctuation imaging (SOFI) is a fluorescence microscopy technique enabling sub-diffraction limit imaging with high temporal resolution by calculating high order statistics of the fluctuating optical signal. The aim of this work is to achieve fast acoustic imaging with enhanced resolution by applying the tools used in SOFI to contrast-enhance ultrasound (CEUS) plane-wave scans. The proposed method was tested using numerical simulations and evaluated using two in-vivo rabbit models: scans of healthy kidneys and VX-2 tumor xenografts. Improved spatial resolution was observed with a reduction of up to 50% in the full width half max of the point spread function. In addition, substantial reduction in the background level was achieved compared to standard mean amplitude persistence images, revealing small vascular structures within tumors. The scan duration of the proposed method is less than a second while current super-localization techniques require acquisition duration of several minutes. As a result, the proposed technique may be used to obtain scans with enhanced spatial resolution and high temporal resolution, facilitating flow-dynamics monitoring. Our method can also be applied during a breath-hold, reducing the sensitivity to motion artifacts.

Functional Flow Patterns and Static Blood Pooling in Tumors Revealed by Combined Contrast-Enhanced Ultrasound and Photoacoustic Imaging.

Alterations in tumor perfusion and microenvironment have been shown to be associated with aggressive cancer phenotypes, raising the need for noninvasive methods of tracking these changes. Dynamic contrast-enhanced ultrasound (DCEUS) and photoacoustic (PA) imaging serve as promising candidates-one has the ability to measure tissue perfusion, whereas the other can be used to monitor tissue oxygenation and hemoglobin concentration. In this study, we investigated the relationship between the different functional parameters measured with DCEUS and PA imaging, using two morphologically different hind-limb tumor models and drug-induced alterations in an orthotopic breast tumor model. Imaging results showed some correlation between perfusion and oxygen saturation maps and the ability to sensitively monitor antivascular treatment. In addition, DCEUS measurements revealed different vascular densities in the core of specific tumors compared with their rims. Noncorrelated perfusion and hemoglobin concentration measurements facilitated discrimination between blood lakes and necrotic areas. Taken together, our results illustrate the utility of a combined contrast-enhanced ultrasound method with photoacoustic imaging to visualize blood flow patterns in tumors. Cancer Res; 76(15); 4320-31. ©2016 AACR.

Extracorporeal acute cardiac pacing by high intensity focused ultrasound.

Ultrasound has been shown to produce Premature Ventricular Contractions (PVC's). Two clinical applications in which acute cardiac pacing by ultrasound may be valuable are: (1) preoperative patient screening in cardiac resynchronization therapy surgery; (2) Emergency life support, following an event of sudden death, caused by cardiac arrest. Yet, previously the demonstrated mean success rate of extra-systole induction by High Intensity Focused Ultrasound (HIFU) in rats is below 4.5% (Miller et al., 2011). This stands in contrast to previous work in rats using ultrasound (US) and ultrasound contrast agents (UCAs), where success rates of close to 100% were reported (Rota et al., 2006). Herein, bi-stage temporal sequences of accentuated negative pressure (rarefaction) and positive pressure HIFU transmission (insonation) patterns were applied to anaesthetized rats under real-time vital-signs monitoring and US imaging. This pattern of insonation first produces a gradual growth of dissolved gas cavities in tissue (cavitation) and then an ultrasonic impact. Results demonstrate sequences of successive successful HIFU pacing. Triggering insonation at different delays from the preceding ECG R-wave demonstrated successful HIFU pacing induction from mid ECG T-wave till the next ECG complex's PR interval. Spatially focusing the beam at different locations allows cumulative coverage of the whole left ventricle. Analysis of the acoustic wave patterns and temporal characteristics of paced PVCs is suggested to provide new insight into the mechanisms of HIFU cardiac pacing. Specifically, the observed HIFU pacing temporal success rate distribution suggests against sarcomere length modulation current being the dominant cellular level mechanism of HIFU cardiac pacing and may allow postulating that membrane deformation currents are dominant at the applied insonation conditions.

Lysyl oxidase-like-2 promotes tumour angiogenesis and is a potential therapeutic target in angiogenic tumours.

Lysyl oxidase-like 2 (LOXL2), a secreted enzyme that catalyzes the cross-linking of collagen, plays an essential role in developmental angiogenesis. We found that administration of the LOXL2-neutralizing antibody AB0023 inhibited bFGF-induced angiogenesis in Matrigel plug assays and suppressed recruitment of angiogenesis promoting bone marrow cells. Small hairpin RNA-mediated inhibition of LOXL2 expression or inhibition of LOXL2 using AB0023 reduced the migration and network-forming ability of endothelial cells, suggesting that the inhibition of angiogenesis results from a direct effect on endothelial cells. To examine the effects of AB0023 on tumour angiogenesis, AB0023 was administered to mice bearing tumours derived from SKOV-3 ovarian carcinoma or Lewis lung carcinoma (LLC) cells. AB0023 treatment significantly reduced the microvascular density in these tumours but did not inhibit tumour growth. However, treatment of mice bearing SKOV-3-derived tumours with AB0023 also promoted increased coverage of tumour vessels with pericytes and reduced tumour hypoxia, providing evidence that anti-LOXL2 therapy results in the normalization of tumour blood vessels. In agreement with these data, treatment of mice bearing LLC-derived tumours with AB0023 improved the perfusion of the tumour-associated vessels as determined by ultrasonography. Improved perfusion and normalization of tumour vessels after treatment with anti-angiogenic agents were previously found to improve the delivery of chemotherapeutic agents into tumours and to result in an enhancement of chemotherapeutic efficiency. Indeed, treatment with AB0023 significantly enhanced the anti-tumourigenic effects of taxol. Our results suggest that inhibition of LOXL2 may prove beneficial for the treatment of angiogenic tumours.

Monitoring radio-frequency thermal ablation with ultrasound by low frequency acoustic emissions--in vitro and in vivo study.

The object of this study was to evaluate the monitoring of thermal ablation therapy by measuring the nonlinear response to ultrasound insonation at the region being treated. Previous reports have shown that during tissue heating, microbubbles are formed. Under the application of ultrasound, these microbubbles may be driven into nonlinear motion that produces acoustic emissions at sub-harmonic frequencies and a general increase of emissions at low frequencies. These low frequency emissions may be used to monitor ablation surgery. In this study, a modified commercial ultrasound system was used for transmitting ultrasound pulses and for recording raw RF-lines from a scan plane in porcine (in vitro) and rabbit (in vivo) livers during radio-frequency ablation (RFA). The transmission pulse was 15 cycles in length at 4 MHz (in vitro) and 3.6 MHz (in vivo). Thermocouples were used for monitoring temperatures during the RFA treatment.In the in vitro experiments, recorded RF signals (A-lines) were segmented, and the total energy was measured at two different frequency bands: at a low frequency band (LFB) of 1-2.5 MHz and at the transmission frequency band (TFB) of 3.5-4.5 MHz. The mean energy at the LFB and at the TFB increased substantially in areas adjacent to the RF needle. These energies also changed abruptly at higher temperatures, thus, producing great variance in the received energy. Mean energies in areas distant from RF needle showed little change and variation during treatment. It was also shown that a 3 dB increase of energy at the low frequency band was typically obtained in regions in which temperature was above 53.3 ± 5° C. Thus, this may help in evaluating regions undergoing hyperthermia. In the in vivo experiments, an imaging algorithm based on measuring the LFB energy was used. The algorithm performs a moving average of the LFB energies measured at segments within the scan plane.Results show that a colored region is formed on the image and that it is similar in size to a measurement of the lesion from gross pathology, with a correlation coefficient of 0.743.

The cardioprotective efficacy of TVP1022 in a rat model of ischaemia/reperfusion.

BACKGROUND AND PURPOSE: Because myocardial infarction is a major cause of morbidity and mortality worldwide, protecting the heart from the ischaemia and reperfusion (I/R) damage is the focus of intense research. Based on our in vitro findings showing that TVP1022 (the S-enantiomer of rasagiline, an anti-Parkinsonian drug) possesses cardioprotective effects, in the present study we investigated the hypothesis that TVP1022 can attenuate myocardial damage in an I/R model in rats. EXPERIMENTAL APPROACH: The model consisted of 30-min occlusion of the left anterior descending artery followed by 4 or 24 h reperfusion. In addition, we investigated the possible mechanisms of cardioprotection in H9c2 cells and neonatal rat ventricular myocytes (NRVM) exposed to oxidative stress induced by H(2) O(2) . KEY RESULTS: TVP1022 (20 and 40 mg·kg(-1) ) administered 5 min before reperfusion followed by an additional dose 4 h after reperfusion reduced the infarct size and attenuated the decline in ventricular function. TVP1022 also attenuated I/R-induced deterioration in cardiac mitochondrial integrity evaluated by mitochondrial swelling capacity. In vitro, using H9c2 cells and NRVM, TVP1022 attenuated both serum free- and H(2) O(2) -induced damage, preserved mitochondrial membrane potential and Bcl-2 levels, inhibited mitochondrial cytochrome c release and the increase in cleaved caspase 9 and 3 levels, and enhanced the phosphorylation of protein kinase C and glycogen synthase kinase-3ß. CONCLUSIONS AND IMPLICATIONS: TVP1022 provided cardioprotection in a model of myocardial infarction, and therefore should be considered as a novel adjunctive therapy for attenuating myocardial damage resulting from I/R injuries.

The combined effect of spatial compounding and nonlinear filtering on the speckle reduction in ultrasound images.

Recently, a spatial compounding ultrasound imaging method was presented that utilizes a conventional 64-element phased array transducer with two unfocused pistons, each placed at one of the sides of the phased array transducer. This method is augmented here by inclusion of nonlinear filtering of the compounded images. The combined effects of the specific spatial compounding and nonlinear filtering on speckle reduction in the generated ultrasound images are studied and evaluated in two stages: First, the image quality is studied when nonlinear filtering is used as part of the spatial compounding. The study is performed by simulations using the Field II program, by processing several B-mode images of a kidney. The second stage compares the results obtained by the simulations to those obtained by in vitro laboratory experiments. Five different compounding strategies and two nonlinear filters, Gaussian and anisotropic diffusion, are investigated and evaluated in terms of image quality parameters-contrast and signal-to-noise ratio. It is shown that the combination of "averaging+nonlinear Gaussian filtering" produces the greatest improvement of image quality. When compared to a conventional phased array imaging system, the spatial compounding method that includes the conventional 64-element phased array transducer with two unfocused pistons, and employs the "averaging+nonlinear Gaussian filtering" strategy, obtains improvement in SNR that has reached 334%. Thus, though this method necessitates a somewhat wider probe, it produces significantly improved images.