Guidelines and good clinical practice recommendations for contrast enhanced ultrasound (CEUS) in the liver--update 2012: a WFUMB-EFSUMB initiative in cooperation with representatives of AFSUMB, AIUM, ASUM, FLAUS and ICUS.

Initially, a set of guidelines for the use of ultrasound contrast agents was published in 2004 dealing only with liver applications. A second edition of the guidelines in 2008 reflected changes in the available contrast agents and updated the guidelines for the liver, as well as implementing some non-liver applications. Time has moved on, and the need for international guidelines on the use of CEUS in the liver has become apparent. The present document describes the third iteration of recommendations for the hepatic use of contrast enhanced ultrasound (CEUS) using contrast specific imaging techniques. This joint WFUMB-EFSUMB initiative has implicated experts from major leading ultrasound societies worldwide. These liver CEUS guidelines are simultaneously published in the official journals of both organizing federations (i.e., Ultrasound in Medicine and Biology for WFUMB and Ultraschall in der Medizin/European Journal of Ultrasound for EFSUMB). These guidelines and recommendations provide general advice on the use of all currently clinically available ultrasound contrast agents (UCA). They are intended to create standard protocols for the use and administration of UCA in liver applications on an international basis and improve the management of patients worldwide.

Initial investigation of acoustic droplet vaporization for occlusion in canine kidney.

Acoustic droplet vaporization (ADV) shows promise for spatially and temporally targeted tissue occlusion. In this study, substantial tissue occlusion was achieved in operatively exposed and transcutaneous canine kidneys by generating ADV gas bubbles in the renal arteries or segmental arteries. Fifteen canines were anesthetized, among which 10 underwent laparotomy to externalize the left kidney and five were undisturbed for transcutaneous ADV. The microbubbles were generated by phase conversion of perfluoropentane droplets encapsulated in albumin or lipid shells in the blood. A 3.5-MHz single-element therapy transducer was aligned with an imaging array in a water tank with direct access to the renal artery or a segmental artery. In vivo color flow and spectral Doppler imaging were used to identify the target arteries. Tone bursts of 1 kHz pulse repetition frequency with 0.25% duty cycle vaporized the droplets during bolus passage. Both intracardiac (IC) and intravenous (IV) injections repeatedly produced ADV in chosen arteries in externalized kidneys, as seen by B-mode imaging. Concurrent with this in two cases was the detection by pulse-wave Doppler of blood flow reversal, along with a narrowing of the waveform. Localized cortex occlusion was achieved with 87% regional flow reduction in one case using IC injections. Vaporization from IV injections resulted in a substantial echogenicity increase with an average half-life of 8 min per droplet dose. Gas bubbles sufficient to produce some shadowing were generated by transcutaneous vaporization of intrarenal artery or IV-administered droplets, with a tissue path up to 5.5 cm.

Rapid 3-D imaging of contrast flow: application in a perfused kidney phantom.

Previous studies indicate imaging of ultrasound contrast in 3-D is potentially superior to 2-D imaging for vascular characterization. A dual-beam, dynamic refill technique, which relies on volumetric contrast clearance and sequential imaging, was used to image a preserved porcine kidney perfused with contrast. A model was developed for the contrast profile across the renal artery to estimate fractional blood volume. This model was used along with refill curve information to measure absolute perfusion within renal cortex for a 100-cm(3) volume. Perfusion measurements from a slice within the volume were also made using a modified interval imaging technique. The measured perfusion using the dual-beam technique was consistent with the perfusion measured using the interval imaging technique (dual-beam values were 1.06 +/- 0.04 x corresponding interval imaging values). These experiments suggest that ultrasound contrast perfusion measurements are independent of the volume of contrast eliminated before refill.

Evolution of bubble clouds induced by pulsed cavitational ultrasound therapy - histotripsy.

Mechanical tissue fractionation can be achieved using successive, high-intensity ultrasound pulses in a process termed histotripsy. Histotripsy has many potential clinical applications where noninvasive tissue removal is desired. The primary mechanism for histotripsy is believed to be cavitation. Using fast-gated imaging, this paper studies the evolution of a cavitating bubble cloud induced by a histotripsy pulse (10 and 14 cycles) at peak negative pressures exceeding 21MPa. Bubble clouds are generated inside a gelatin phantom and at a tissue-water interface, representing two situations encountered clinically. In both environments, the imaging results show that the bubble clouds share the same evolutionary trend. The bubble cloud and individual bubbles in the cloud were generated by the first cycle of the pulse, grew with each cycle during the pulse, and continued to grow and collapsed several hundred microseconds after the pulse. For example, the bubbles started under 10 microm, grew to 50 microm during the pulse, and continued to grow 100 microm after the pulse. The results also suggest that the bubble clouds generated in the two environments differ in growth and collapse duration, void fraction, shape, and size. This study furthers our understanding of the dynamics of bubble clouds induced by histotripsy.

Standardization of power Doppler quantification of blood flow in the human fetus using the aorta and inferior vena cava.

OBJECTIVE: This study aimed to test the applicability to the fetus of mathematical techniques developed to standardize power Doppler indices of adult regional perfusion. METHOD: Fetal power Doppler imaging was performed on 14 fetuses (25-37 weeks' gestation) using a standardized parasagittal plane, examining renal and aortic blood flow, and additionally inferior vena caval (IVC) flow in one fetus. Images were stored and transferred for off-line computer analysis using purpose-designed software. We first tested the need for techniques to remove the effects of red blood cell clumping on power Doppler amplitude, then performed further analyses to: (1) investigate cardiac cycle effects on aortic amplitude; (2) determine the spatial consistency of, and influence of angle of insonation on, maximal pixel value; (3) quantify temporal consistency; and (4) compare peak pixel values in the fetal aorta and IVC. RESULTS: No rouleaux effect on the vascular profile was detectable, in contrast to that identified in the adult. Within each fetus a consistent value was seen in the center of the aorta corresponding to 100% vascular amplitude, which was unaltered by the phase of the cardiac cycle, with a coefficient of variation of 28.9% at 89 degrees and 6.5% at 73 degrees . This value was constant in the aorta and IVC. CONCLUSION: Fetal blood does not appear to form rouleaux to any significant degree, so there is no artificially elevated maximal value for power Doppler amplitude as seen in the adult. We propose that the value representing 100% amplitude may be consistently measured in the center of large fetal vessels such as the aorta, allowing the direct measurement of fractional moving blood volumes in the human fetus.

Analysis of refill curve shape in ultrasound contrast agent studies.

Contrast destruction and replenishment by Flash Echo Imaging (FEI) (also referred to as interval or intermittent imaging) has been qualitatively and quantitatively used for tissue blood refill measurements. Many features and capabilities of contrast refill in tissue blood flow and perfusion remain to be elucidated. To aid the development and full reliable utilization of the technique in medical practice, in this paper we undertake physical and mathematical modeling to evaluate different measures derivable from FEI and to provide a basis for the further study of sensitivity and stability of such measures for the detection and measurement of various flow properties and abnormalities. A phantom was developed and used to conduct a dynamic contrast study. Refill curves were investigated as a means of calculating the mean transit time (MTT) and investigating other information that can be determined from their shape. Exponential and error function fits and the area above these curves were used to estimate MTT. The bubble disruption zone was visually measured and theoretically modeled. Computer simulated refill curves based on the flow phantom for different velocity ranges were then computed and compared to the experimental refill curves. The simulated refill curves closely matched the experimental curves in both shape and MTT. The simulated refill curves matched the shape of the experimental results for different velocity ranges. Another simulation examined how a real circulatory system might influence refill. Different refill curve shapes were obtained for different vascular models. Models including the large arteries and veins showed a much faster initial slope than models where the large vessels were not included. Likewise, simulated "shunting" displayed a different slope than models without "shunting" and specific portions of the refill curve that could maximally distinguish shunting. This computer simulation could lead to some experimental hypotheses about differences between normal and cancerous blood flow.

Decrease in fibrin content of venous thrombi in selectin-deficient mice.

The purpose of this study was to quantify the fibrin content of thrombi produced in a mouse model of venous thrombosis and correlate this to thrombus mass. The role of P-selectin, E-selectin, and IL-10 on thrombus fibrin content was analyzed using knockout (KO) mice. Five groups of mice were evaluated: control (N = 10), P-selectin KO (N = 7), E-selectin KO (N = 5), combined E-/P-selectin KO (N = 12), and IL-10 KO (N = 10). Venous thrombosis was induced by ligation of the infrarenal IVC. Mice were sacrificed on postoperative days (POD) 2 and 6. Thrombus mass was calculated. Sections of IVC were stained with an antibody that cross reacts with mouse fibrin. The distribution of RGB color pixels was generated from digitized micrographs of the thrombus of each animal. The mean pixel value for each group was compiled and analyzed using 2-way ANOVA. Mean pixel value per group was correlated with the mean thrombus mass per group. Color analysis demonstrated significant decreases in the analyzed fibrin content on POD-2 between the control vs E-/P-selectin KO (P < 0.05) and control vs IL-10 KO (P < 0.05) groups. In addition, significantly less fibrin staining was noted on POD-6 between the control vs E-selectin KO (P = 0.03), control vs P-selectin KO (P = 0.01), and control vs E-/P-selectin KO (P < 0.01). There was a strong overall correlation between the mean pixel value for each group and the thrombus mass (R = 0.964; P < 0.01). This study demonstrates a difference in fibrin content of thrombi produced in animals deficient in E-selectin, P-selectin, and IL-10, supporting their importance in thrombus amplification, fibrin formation, and the mass of thrombus formed.

Doppler quantitative measures by region to discriminate prostate cancer.

This study was conducted to assess if sonographic discrimination between healthy and cancerous prostate tissue might be improved using regional analysis of ultrasound (US) Doppler measures. A total of 39 subjects underwent 3-D Doppler sonography before radical prostatectomy. Cancer locations were identified from hematoxylin and eosin (H&E)-stained slides. Three prostate data volumes consisting of a frequency shift and power-mode Doppler US and whole mount histology images were spatially registered for each prostate, then divided into entirely 1 mL-sized regions of cancerous or noncancerous tissue. Each prostate was visually divided into a peripheral and a periurethral region within which US Doppler measures were calculated. Receiver operating characteristic (ROC) and simulated biopsy analyses within each prostate were performed. Mean speed in colored pixels (V), and speed-weighted pixel density (SWD) are good discriminators for prostate cancer in the periurethral and the peripheral regions, respectively. Using SWD in a simulated biopsy yields increased cancer detection in the peripheral region.

Analysis of three-dimensional Doppler ultrasonographic quantitative measures for the discrimination of prostate cancer.

OBJECTIVE: The purpose of this study was to determine whether several quantitative ultrasonographic measures have potential to discriminate prostate cancer from normal prostate and to determine the best combination of these measures. The true spatial distributions of cancer within the prostates studied were obtained histologically after radical prostatectomy. The relationship between Doppler ultrasonography and microvessel count was also investigated. METHODS: Three-dimensional Doppler ultrasonographic data were acquired from 39 patients before radical prostatectomy. The removed prostate was sectioned, and whole-mount hematoxylineosin-stained slides were used to identify all regions of cancer within each prostate. These histologic and ultrasonographic data were spatially registered. Doppler ultrasonographic measures were calculated within uniformly sized three-dimensional regions that were either entirely cancerous or noncancerous, and receiver operating characteristic analysis was performed on the results. Microvessel counts were made within each contiguous cancerous region and correlated with ultrasonographic measures. RESULTS: Color pixel density was the best simple measure for discriminating prostate cancer (accuracy, 80%). The mean power mode value (normalized mean power in color pixels) was inversely related to cancer with an accuracy of 1--normalized mean power in color pixels = 65% (low mean power is more cancerous). When color pixel density was combined with the normalized mean power in color pixels, its accuracy improved slightly to 84%. The peak microvessel count had a negative correlation with color pixel density as well as with cancer stage. CONCLUSION: Doppler ultrasonography does provide discriminatory information for prostate cancer, with color pixel density being the most promising measure.

Interleukin-8 administration enhances venous thrombosis resolution in a rat model.

BACKGROUND: Therapy for deep vein thrombosis (DVT) resolution in those patients in whom a complication or contraindication to anticoagulation occurs is limited. As prior work suggests that thrombus maturation involves early influx of neutrophils (PMN) and neovascularization, we hypothesized that administering the proinflammatory/proangiogenic chemokine interleukin (IL)-8 might accelerate thrombus resolution. MATERIALS AND METHODS: An established rodent model of DVT (inferior vena cava [IVC] ligation) was used whereby daily intravenous recombinant human IL-8 (1 microg) or vehicle control was administered, with sacrifice at 4 and 8 days. Prior to sacrifice and at harvest, duplex ultrasound of the DVT and femoral venous pressure measurements were performed. Thrombi were analyzed by immunohistochemical techniques for PMN, monocytes, and neovascularization; for chemokines, by enzyme-linked immunoassay; and fibrosis, by hydroxyproline assay and trichrome staining. RESULTS: IL-8 accelerated thrombus dissolution 4 days after IVC ligation, with 6-fold increased thrombus blood flow by duplex ultrasound and a 23% increased absolute femoral venous pressure compared with controls (both P < 0.05). These findings may be partially explained by the fact that animals receiving IL-8, as compared with controls, had 2.5-fold greater thrombus neovascularization (with a trend continuing to 8 days) and increased PMN at 4 days. Thrombus vascular endothelial growth factor was significantly reduced at 8 days postligation, while monocyte chemotactic protein-1 and macrophage inflammatory protein-1alpha were not altered by IL-8 administration. At 8 days post-IVC-ligation, fibrosis was 12-fold greater with IL-8 treatment compared with controls. CONCLUSIONS: A proinflammatory/proangiogenic thrombus milieu, as conferred by IL-8, enhances thrombus resolution and underscores the important relationship between neovascularity and inflammation.

Analysis of three-dimensional ultrasound Doppler for the detection of prostate cancer.

OBJECTIVES: To investigate the relative effectiveness of Doppler ultrasound quantitative measures in discriminating prostate cancer from normal prostate tissue. The true locations of prostate cancer within these prostates were determined by histologic examination after radical prostatectomy. METHODS: Three-dimensional Doppler ultrasound data were acquired from 39 men before radical prostatectomy. The removed prostates were sectioned and all cancerous regions in each prostate were identified on whole-mount hematoxylin-eosin-stained slides. The ultrasound and histologic data were then spatially registered. Biopsy results were simulated on a grid of potential sites within each prostate. Along each simulated biopsy site, the amount of cancer was computed from the hematoxylin-eosin-identified cancerous regions and the peak speed-weighted pixel density (SWD) was compared. RESULTS: By selecting the biopsy sites with higher associated SWDs within each sextant, the probability of having at least one positive biopsy within a prostate increased from 75% if the SWD was ignored to 85% if only the top 15% of potential biopsy sites in each sextant were selected. This trend was seen within each sextant individually as well. CONCLUSIONS: Doppler ultrasound provides discriminatory information for prostate cancer using the SWD. Translating this into a practical strategy that might improve the yield of prostate biopsy remains under development. The results of our study indicate that biopsying regions of high Doppler color could potentially increase the cancer yield to a small degree and improve the accuracy of the biopsy results. These results also objectively verify previous visual studies suggesting a modest improvement with the use of color Doppler.

Discrimination of sonographically detected breast masses using frequency shift color Doppler imaging in combination with age and gray scale criteria.

Frequency shift color Doppler imaging was assessed in conjunction with patient age and gray scale (GS) features for discriminating benign from malignant breast masses. Thirty-eight women with sonographically detected masses scheduled for biopsy were evaluated using a 6- to 13-MHz scan head, and the masses were delineated in ultrasonographic image volumes. Vascularity in and around each mass was quantified using speed-weighted pixel density (SWD). Gray scale features were ranked visually on a linear scale. Combinations of indices were compared with histologic findings (18 benign and 20 malignant). Receiver operating characteristic analysis ranked performance in decreasing order from the SWD-Age-GS index, to SWD-GS, SWD-Age, Age-GS, GS criteria, SWD, and Age. At 100% sensitivity, SWD-Age-GS, SWD-GS, and SWD-Age discriminated benign from malignant masses with specificities of 94%, 89%, and 72%, respectively. These results indicate significant improvement in ultrasonographic discrimination of sonographically detected breast masses by combining the vascularity measure SWD with age and GS criteria.

Volume flow measurement using Doppler and grey-scale decorrelation.

A technique for volumetric blood flow measurement was developed by combining standard Doppler measurements with grey-scale decorrelation. Steered Doppler is used to determine the in-plane velocities, which are then used to extract the out-of-plane velocities from the temporal A-line decorrelation. As a result, a three-dimensional (3-D) vector flow field can be computed over the imaging plane using a single clinical transducer without knowledge of the vessel orientation. Volume flow is computed by integrating the out-of-plane flow over the vessel cross-section. The algorithm was tested using a scattering-enhanced fluid in a 6.4-mm diameter dialysis tubing. For a wide range of transducer angles, the volume flow was accurately measured to within 28% in these preliminary tests.

Doppler US gating of cardiac MR imaging.

RATIONALE AND OBJECTIVES: Electrocardiographic (ECG) gating of cardiac magnetic resonance (MR) imaging has been problematic for many reasons. The purpose of this study was to demonstrate the feasibility of using Doppler ultrasound (US) gating, either directly off the moving cardiac wall or the systolic upstroke of the arterial signal from the great vessels in neck, in alternative gating modes. MATERIALS AND METHODS: A 2.5-MHz, range-gated Doppler US device was used with A-mode guidance for gating directly off left ventricular wall motion. A 4- or 8.1-MHz, continuous-wave (CW) Doppler US device was used for gating off the systolic upstroke from the great vessels in the neck. The subject undergoing imaging held the transducer against his chest for range-gated Doppler US and against his neck for 8.1-MHz CW Doppler US. The 4-MHz transducer was strapped to the subject's neck. Modified Doppler signals were fed back into the gating circuitry of the MR imager to achieve cardiac synchrony. RESULTS: Cardiac gating was achieved by using both the range-gated technique directly off the cardiac wall and the CW method off blood flow from the great vessels. Problems occurred with radiofrequency shielding during the range-gated method; however, these problems were almost completely removed by use of the CW Doppler probes. CONCLUSION: Doppler US gating of MR images is possible and potentially could overcome many shortcomings of ECG gating. Subsequent embodiments of the technique will require improved radiofrequency shielding in the range-gated technique.

Acoustic droplet vaporization for therapeutic and diagnostic applications.

A phase shift droplet emulsion is introduced as an aid to unusual ultrasound (US) applications. The transpulmonary droplet emulsion (90% < 6 microm diameter) is made by mixing saline, bovine albumin and dodecafluoropentane. It has been observed that an acoustic pressure threshold exists, above which the droplets vaporize into bubbles approximately 25 times the original diameter. For frequencies between 1.5 and 8 MHz, the threshold decreases from 4.5 to 0.75 MPa peak rarefactional pressure. This paper presents preliminary results for droplet preparation and their evaporation as a function of applied acoustic pressure and frequency, as well as simulations of the lifetime of these gas bubbles based on gas diffusion. In vivo experiments were simulated by the evaporation of droplets in blood flowing under attenuating material. We propose that this agent might be useful for tissue occlusion in cancer treatment, as well as for phase aberration corrections in acoustic imaging.

Cavitation nucleation agents for nonthermal ultrasound therapy.

The use of a nucleation-promoting agent can greatly enhance therapeutically useful nonthermal bioeffects. A blank agent (saline), Optison ultrasound contrast agent, a stabilized perfluoropentane droplet suspension (SDS), and retained air space were compared as nucleation agents in whole blood. Fresh canine whole blood with added agent was exposed in 1.3-ml disposable pipette bulbs to lithotripter shock waves (2-Hz rate; +24.4, -5.2 MPa peak pressure amplitudes). Cavitation activity was assessed by measuring hemolysis. The droplet suspension performed nearly as well as retained air when added at a concentration sufficient to provide a roughly equal volume of gas after vaporization. Optison also yielded nucleation, but a concentration of 10%-20% was needed for large enhancement of hemolysis comparable to 5% SDS. Exposure at room temperature, which was less than the 29 degrees C boiling point of perfluoropentane, eliminated the enhancement of the hemolysis effect relative to the blank. Application of 100-kPa excess pressure during exposure reduced but did not eliminate the nucleation ability of Optison, SDS, or retained air. However, this small pressure (relative to the peak positive pressure of the shock waves) eliminated the hemolysis induced with the blank agent. The stabilized perfluoropentane droplet suspension appears to be a good nucleation agent for nonthermal ultrasound therapy applications.

3D spatial compounding of ultrasound images using image-based nonrigid registration.

Medical ultrasound images are often distorted enough to significantly limit resolution during compounding (i.e., summation of images from multiple views). A new, volumetric image registration technique has been used successfully to enable high spatial resolution in three-dimensional (3D) spatial compounding of ultrasound images. Volumetric ultrasound data were acquired by scanning a linear matrix array probe in the elevational direction in a focal lesion phantom and in a breast in vivo. To obtain partly uncorrelated views, the volume of interest was scanned at five different transducer tilt angles separated by 4 degrees to 6 degrees. Pairs of separate views were registered by an automatic procedure based on a mutual information metric, using global full affine and thin-plate spline warping transformations. Registration accuracy was analyzed automatically in the phantom data, and manually in vivo, yielding average registration errors of 0.31 mm and 0.65 mm, respectively. In the vicinity of the warping control points, registrations obtained with warping transformations were significantly more accurate than full affine registrations. Compounded images displayed the expected reduction in speckle noise and increase in contrast-to-noise ratio (CNR), as well as better delineation of connective tissues and reduced shadowing. Compounding also revealed some apparent low contrast lobulations that were not visible in the single-sweep images. Given expected algorithmic and hardware enhancements, nonrigid, image-based registration shows great promise for reducing tissue motion and refraction artifacts in 3D spatial compounding.

Speckle decorrelation flow measurement with B-mode US of contrast agent flow in a phantom and in rabbit kidney.

PURPOSE: To use speckle decorrelation in the presence of ultrasonographic (US) contrast agent as an alternative flow measurement technique to Doppler US. MATERIALS AND METHODS: In vivo and in vitro studies were performed. A tube with flowing saline solution containing contrast agent was positioned horizontally across a US image. The amount of decorrelation between a series of images was recorded. The flow profile across the tube was generated by averaging the decorrelation values and was compared with a Doppler frequency shift image. In addition, B-mode images of six rabbit kidneys were obtained during and after intravenous injection of contrast agent. Images were analyzed to compute the correlation between successive points in time. RESULTS: The velocity profiles across the tube were parabolic, with the fastest flow rates measured in the center of the tube. In the rabbit kidneys, measurements indicated the largest decorrelation rates occurred in the larger vessels. The cortical decorrelation rates were significantly slower than those for the hilar vessels (P < .05) and were relatively angle independent. CONCLUSIONS: Decorrelation flow measurements can be used to estimate flow in vitro and in vivo similar to measurements obtained with Doppler US but with less angle dependence. These measurements could lead to a US perfusion technique.

Interlaboratory comparison of ultrasonic backscatter, attenuation, and speed measurements.

In a study involving 10 different sites, independent results of measurements of ultrasonic properties on equivalent tissue-mimicking samples are reported and compared. The properties measured were propagation speed, attenuation coefficients, and backscatter coefficients. Reasonably good agreement exists for attenuation coefficients, but less satisfactory results were found for propagation speeds. As anticipated, agreement was not impressive in the case of backscatter coefficients. Results for four sites agreed rather well in both absolute values and frequency dependence, and results from other sites were lower by as much as an order of magnitude. The study is valuable for laboratories doing quantitative studies.