Kristine G. Baker

Contrast-enhanced B-mode US angiography in the assessment of experimental in vivo and in vitro atherosclerotic disease

RATIONALE AND OBJECTIVES This study was performed to (a) test the hypothesis that filling the arterial lumen with echoes at B-mode ultrasound (US) enables the assessment of wall and luminal abnormalities and (b) compare contrast material-enhanced B-mode US with color and power Doppler US angiography. MATERIALS AND METHODS Atherosclerotic lesions were created in 14 rabbit aortas and imaged with color Doppler and B-mode US before and after the intravenous administration of 0.3 mL of AF0150, a US contrast agent. In addition, four replicas of diseased human carotid arteries were immersed in a tissue-mimicking phantom and imaged with B-mode and color and power Doppler US before and after the administration of 1 mL of AF0150 per liter of porcine blood. Radiopaque plastic casts of the rabbit aortas and contact radiographs of the plastic replicas served as standards. RESULTS Although color and power Doppler US allowed immediate localization of the lumen, precise estimation of stenoses and reliable visualization of surface irregularities were not possible. After AF0150 administration, angiogram-like images of the lumen were created with B-mode US, allowing rapid assessment of the entire vessel lumen and wall. Consequently, luminal stenoses were more accurately measured than with unenhanced B-mode US (r2 = 0.94, P < .0001 vs r2 = 0.21, P = .25) or Doppler (r2 = 0.42, P < .03). In addition, plaques and ulcerations were visible only with contrast-enhanced B-mode US. CONCLUSION Microbubbles fill the arterial lumen with echoes at B-mode US, creating an angiogram-like image. The ability to visualize the inner and outer surfaces of the vascular wall improved the evaluation of luminal and wall abnormalities.

Comparison of standard and second harmonic B-mode sonography in the detection of segmental renal infarction with sonographic contrast in a rabbit model.

This study compares contrast‐enhanced fundamental and second harmonic B‐mode sonography using a rabbit renal infarct model. Segmental renal infarctions were produced in 13 rabbits by embolizing a 0.7 mm bead into the renal artery 1 day prior to imaging. An ultrasonographic unit equipped with an L10‐5 transducer and second harmonic imaging capability was used. Real‐time recordings were made during the injection of 0.5 ml of an experimental formulation of a perfluorohexane vapor‐stabilized microbubble (AF0145) given into the ear vein, and the imaging technique alternated between standard and harmonic imaging every 20 s. Each rabbit received two injections 1 h apart. To control for the effect of peak bolus enhancement, the initial imaging technique used for the first injection was randomized, and the other technique was used initially for the second injection. The videointensity difference between the infarcted and the normal cortex was then calculated and evaluated as a function of time. The infarcted segment could not be seen before administration of contrast agent with either technique. Although the infarction could be seen after injection of contrast agent with either technique, image contrast and contrast duration were nearly 75% greater for the harmonic technique than for the standard technique. AF0145 allows the visualization of segmental renal infarction on standard B‐mode imaging. The second harmonic B‐mode technique significantly increases image contrast and contrast duration.

Effect of acquisition rate on liver and portal vein enhancement with microbubble contrast

We showed that tissue enhancement with microbubbles is dependent upon transmit power. Because intermittent imaging decreases bubble exposure to ultrasound, and also decreases the ability of the sonographer to maintain anatomic orientation, we aimed to determine the optimum frame rate that maximizes enhancement and allows for continued anatomic orientation. Seven rabbits with an avascular liver lesion created by percutaneous injection of 1 mL ethyl alcohol 7 days earlier were imaged with an Acuson 128XP/10 using a 7-MHz sector transducer at fixed transmit power. Each rabbit was imaged 5 times in random order, at 1 frame/30 s, 1frame/5 s, 1frame/s, 4 frames/s, and 28 frames/s. The same plane was imaged at all frame rates from before to 15 min after the bolus injection of 0.3-mL (0.1-0.12 mL/kg) of AF0150 (Imagent, Alliance Pharmaceutical Corp., San Diego, CA). Liver and portal vein videointensity relative to the lesion were evaluated over time. In this study, liver enhancement progressively increased as the frame rate was reduced (p<0.001). Peak, duration, and area under the time-intensity curve were all greater at the lower frame rates (1 fr/30 s, 1 fr/5 s, and 1 fr/s) than at 28 fr/s (p<0.05). Anatomic orientation was maintained at 1 frame/s rate at which peak enhancement was 44% greater and duration was 100% longer than at 28 frames/s (p<.05). Portal vein enhancement was not affected by frame rate. In conclusion, with intermittent imaging, enhancement was dependent upon frame rate and the ability of the region being imaged to replenish its bubbles between consecutive acquisitions. The 1 frame/s allowed for anatomic orientation and adequate tissue contrast.

Renal resistive index in experimental partial and complete ureteral obstruction.

Rationale and objectives Recent clinical work suggests that the Doppler resistive index (RI) may be useful in distinguishing obstructive from nonobstructive hydronephrosis. We evaluated the usefulness of the RI in a rabbit model of hydronephrosis. Methods Unilateral partial ureteral obstruction was produced in nine rabbits and complete obstruction in another nine. Three sham operations were performed, and these animals served as control subjects. The RI was measured in all kidneys before and 6 hr after surgery and on days 1, 4, and 7 postoperatively. The RI and the difference in RI (delta RI) between the obstructed and normal kidney were evaluated over time using a two-way analysis of variance. The intravenous urography and Whitaker tests served as gold standards. Results Hydronephrosis was observed on sonograms in all obstructed kidneys. Comparing groups, there was no significant difference in mean RI or delta RI between the three groups at any time point. Looking at individual groups over time, there was no significant change in mean delta RI, whereas the change in mean RI was significantly elevated above baseline only in the complete obstruction group at 6 hr (p = .002) and on days 4 (p = .008) and 7 (p = .006). In evaluating varying thresholds of RI and delta RI, we could not consistently discriminate between normal and obstructed kidneys. Conclusion Although complete obstruction caused a significant increase in RI, partial obstruction failed to do so. RI and delta RI values proved to be insensitive predictors of obstruction in this rabbit model.

B-Mode Enhancement of the Liver with Microbubble Contrast Agent

RATIONALE AND OBJECTIVES The purpose of this study was to evaluate the accuracy of contrast material-enhanced sonography in the detection of liver lesions by using an animal model. MATERIALS AND METHODS A total of 36 rabbits, 12 normal and 24 with one, two, or more VX2 tumors implanted percutaneously, were imaged on an Acuson 128XP/10 with a 7-MHz sector transducer by a sonographer blinded to the study assignments. The sonographer assigned rabbits to four groups (no, one, two, more than two tumors) based on the number of lesions detected before and then after the intravenous bolus injection of 0.5 mL of AF0150. S-VHS video segments or pre- and postcontrast images were separated, randomized, and evaluated by a blinded reader. Necropsy served as the gold standard. RESULTS Classification of rabbits as normal or tumor bearing on the precontrast images produced three false-positive results and three false-negative results for the blinded sonographer and six false-positive results and two false-negative results for the blinded reader. On postcontrast images, all rabbits were correctly classified by both observers. The correlation of the classification of whether rabbits had no, one, two, or more tumors relative to the pathologic classification on precontrast images was poor to fair (K = 0.349 +/- 0.099 for the sonographer and 0.274 +/- 0.111 for the reader), whereas the postcontrast correlation was good to excellent (K = 0.924 +/- 0.099 for the sonographer and 0.809 +/- 0.076 for the reader). CONCLUSION AF0150 markedly increased the ability of the sonographer and the blinded reader to distinguish normal from tumor-bearing animals and improved the classification of rabbits with more than one liver tumor.

Assessment of liver and kidney enhancement with a perfluorocarbon vapor-stabilized US contrast agent

RATIONALE AND OBJECTIVES The authors evaluated the time-echogenicity response of liver, kidney, and implanted VX2 tumor after injection of a microbubble contrast medium and assessed use of an avascular lesion as an internal standard. MATERIALS AND METHODS Twenty-one New Zealand White rabbits were studied. To evaluate use of an internal standard and the dose-response relationship, nine rabbits with 7-day-old avascular liver lesions created by alcohol ablation received 0.1, 0.25, 0.5, and 1.0 mL of AF0145, a microbubble contrast agent. To evaluate tumor echogenicity, 12 rabbits implanted with VX2 tumor in the liver (six also underwent alcohol ablation) received 0.5 mL of AF0145. Videodensitometry was used to analyze echogenicity changes over 10 minutes. RESULTS Echogenicity of the alcohol-ablated liver was not affected by contrast material administration. Liver and kidney echogenicity relative to ablation increased linearly with dose, peaking 1 minute after injection and decaying to baseline over 9 minutes. Contrast material administration defined the size and margins of VX2 lesions more clearly. In the arterial phase, the tumor rim was hyperechoic relative to surrounding liver, becoming isoechoic during the portal venous phase then hypoechoic during the late phase parenchymal phase. CONCLUSIONS Lesions created by alcohol ablation can be used as an internal standard for quantitative analysis of adjacent tissues. AF0145 enhances perfused tissues, including vascular tumors, at gray-scale, real-time ultrasonography and enhances the liver.

Effect of ultrasound transmit power on liver enhancement with Imagent® US, a PFC‐stabilized microbubble contrast agent

The pressure of the ultrasound wave may limit the longevity of microbubble‐based contrast agents. This study evaluated liver enhancement over time as a function of transmit power after the administration of AFO145 (Imagent® US; Alliance Pharmaceutical Corp., San Diego, CA). Eight rabbits with an avascular liver lesion created by percutaneous injection of 1.0 ml of ethyl alcohol 7 days prior to scanning were imaged with an Acuson 128XP/10 at 7 MHz before and after four separate intravenous injections of 0.25 ml of AFO145 spaced at least 1 h apart. The avascular lesion served as an internal standard against which liver enhancement could be compared. After contrast injection, scanning over the same plane was either continuous at (a) maximum or (b) minimum transmit power (9 dB below maximum), or intermittent at (c) minimum power for 5 s every 15 s, or (d) for 5 s every 60 s. Each session was terminated after 15 min or when contrast was no longer visible in the hepatic parenchyma and blood vessels. Videodensitometry was used to assess liver‐to‐lesion intensity difference over time. Both the degree and duration of liver enhancement were dependent on the transmit power. Liver enhancement with imaging at minimum power for 5 s/min was nearly two times greater and persisted nearly eight times longer (P < 0.01) than at maximum power and continuous insonation. Ultrasound transmit power affects both the peak and duration of liver enhancement. A lower power and shorter insonation time after AFO145 administration dramatically lengthens the imaging window for liver lesion detection.

Liver-tumor detection with ultrasound contrast: A blinded prospective study in rabbits

Ultrasound has evolved into a frontline modality in most applications except for the detection of focal liver disease. Despite the wide availability, portability, relatively low cost, and high patient acceptance of ultrasound, CT and MR are currently considered the standards for detection and characterization of focal liver lesions (1). The superiority of MR and particularly CT is in part due to the availability of IV contrast material. Contrast agents promote lesion visualization by increasing image contrast owing to the differences in their wash-in and washout rates between normal and abnormal tissues. Others and we have demonstrated that ultrasound contrast agents can provide gray-scale enhancement of vessels and tissues, including those of the liver (2-5). We have also shown that the contrast agent can be observed filling the hepatic arteries, the portal veins, and then the hepatic parenchyma in real time (5). This triple-phase enhancement pattern increases the conspicuity of vascular tumors that become brighter than liver during the arterial phase, potentially expanding the role of sonography in tumor diagnosis and characterization. Proof that resection of colorectal liver metastases increases survivorship heightens the need not only to recognize their presence, but also to detect all lesions. Since the number and size of metastases determines resectability (1),

Efficacy of Imagent BP at 1.5ml/kg in a rabbit liver tumor model.

We have shown using a Vx2 rabbit model that 3 and 5ml/kg of perflubron emulsion were highly effective in imaging liver tumors. The results from preliminary clinical trials suggested that should the infusion rate be increased, a 1.5ml/kg may be efficacious. This study determined whether 1.5ml/kg given as a bolus IV would be efficacious to image liver tumors using a Vx2 rabbit model. Vx2 tumors were implanted in 5 NZW rabbits, CT of the liver was performed during held expiration at 80kV and 800mAS, before and shortly after 1.5ml/kg Imagent BP (ImBP), again at 30 minutes and 3 days. Regions of interest (ROIs) were drawn over the CT image of the spleen, liver, inferior vena cava, and tumor, CT# obtained and average enhancement of each tissue calculated at each time point. 4 animals had tumors .5cm or greater. Precontrast, tumors were faintly seen on CT. Blood was brighter than liver shortly after infusion and isointense with liver at 30 minutes. Tumors did not enhance following contrast. Except for the liver and spleen, all tissues returned to baseline on the 3rd day. Therefore a clinical trial to determine the efficacy of 1.5ml/kg ImBP to image the liver is warranted.

Renal Resistive Index in Experimental Partial and Complete Ureteral Obstruction

RATIONALE AND OBJECTIVES Recent clinical work suggests that the Doppler resistive index (RI) may be useful in distinguishing obstructive from nonobstructive hydronephrosis. We evaluated the usefulness of the RI in a rabbit model of hydronephrosis. METHODS Unilateral partial ureteral obstruction was produced in nine rabbits and complete obstruction in another nine. Three sham operations were performed, and these animals served as control subjects. The RI was measured in all kidneys before and 6 hr after surgery and on days 1, 4, and 7 postoperatively. The RI and the difference in RI (delta RI) between the obstructed and normal kidney were evaluated over time using a two-way analysis of variance. The intravenous urography and Whitaker tests served as gold standards. RESULTS Hydronephrosis was observed on sonograms in all obstructed kidneys. Comparing groups, there was no significant difference in mean RI or delta RI between the three groups at any time point. Looking at individual groups over time, there was no significant change in mean delta RI, whereas the change in mean RI was significantly elevated above baseline only in the complete obstruction group at 6 hr (p = .002) and on days 4 (p = .008) and 7 (p = .006). In evaluating varying thresholds of RI and delta RI, we could not consistently discriminate between normal and obstructed kidneys. CONCLUSION Although complete obstruction caused a significant increase in RI, partial obstruction failed to do so. RI and delta RI values proved to be insensitive predictors of obstruction in this rabbit model.