Resting coronary flow drives the daily pattern in coronary flow reserve in patients with chest pain without obstructive epicardial stenosis.

Objectives: Ischemia with no obstructive coronary artery disease (INOCA) is a risk factor for major adverse cardiovascular events and is characterized by abnormal coronary microvascular tone. In patients with INOCA, adverse cardiovascular events most commonly occur in the morning compared to other times of the day and night. Materials and methods: We tested whether coronary microvascular function varies diurnally with attenuation in the morning in patients with symptomatic coronary artery disease without significant (>50%) epicardial stenosis. We evaluated data from 17 patients studied in the AM (700-1159 h) and 11 patients in the PM (1200-1800 h). Coronary microvascular function was measured using perfusion contrast imaging at rest and after infusion of intravenous regadenoson. We calculated microvascular flow reserve as the ratio of hyperemic to resting flow. Along with independent sample t-tests, we performed bootstrapping procedures to test mean differences between AM and PM groups, using the bias-corrected and accelerated method with 5,000 bootstrapped samples. Results and conclusion: The AM and PM groups were matched for demographic and existing risk factors. Coronary microvascular flow reserve was ∼33% higher in the AM compared to the PM (P = 0.025, BCa 95% CI [0.25, 1.64]; Hedge's g = 0.89, 95% CI [0.11, 1.66]) as a result of significantly lower resting flow (∼50%) in the AM compared to the PM (P = 0.03, M Diff = -56.65, BCa 95% CI [-118.59, -2.12]; Hedge's g = -0.86, 95% CI [-1.60, -0.06]). Our observations are of clinical value and can influence diagnosis and treatment in the clinic based on the time of day of measurements.

Changes in microvascular perfusion of heart and skeletal muscle in sheep around the time of birth.

NEW FINDINGS: What is the central question of this study? How does the microvascular perfusion of striated muscle change during the dynamic developmental period between the late gestation fetus and early neonate? What is the main finding and its importance? In both myocardium and skeletal muscle, perfusion of striated muscle is significantly reduced in the neonate compared to the late term fetus, but flow reserve is unchanged. The results suggest striated muscle capillary networks grow more slowly relative to the myofibres they nourish during the perinatal period. ABSTRACT: Microvascular perfusion of striated muscle is an important determinant of health throughout life. Birth is a transition with profound effects on the growth and function of striated muscle, but the regulation of microvascular perfusion around this transition is poorly understood. We used contrast-enhanced ultrasound perfusion imaging (CEUS) to study the perfusion of left ventricular myocardium and hindlimb biceps femoris, which are populations of muscle with different degrees of change in pre- to postnatal workloads and different capacities for postnatal proliferative growth. We studied separate groups of lambs in late gestation (135 days' gestational age; 92% of term) and shortly after birth (5 days' postnatal age). We used CEUS to quantify baseline perfusion, perfusion during hyperaemia induced by adenosine infusion (myocardium) or electrically stimulated unloaded exercise (skeletal muscle), flow reserve and oxygen delivery. We found heart-to-body weight ratio was greater in neonates than fetuses. Microvascular volume and overall perfusion were lower in neonates than fetuses in both muscle groups at baseline and with hyperaemia. Flux rate differed with muscle group, with myocardial flux being faster in neonates than fetuses, but skeletal muscle flux being slower. Oxygen delivery to skeletal muscle at baseline was lower in neonates than fetuses, but was not significantly different in myocardium. Flow reserve was not different between ages. Given the significant somatic growth, and the transition from hyperplastic to hypertrophic myocyte growth occurring in the perinatal period, we postulate that the primary driver of lower neonatal striated muscle perfusion is faster growth of myofibres than their associated capillary networks.

Augmentation of Tissue Perfusion with Contrast Ultrasound: Influence of Three-Dimensional Beam Geometry and Conducted Vasodilation.

BACKGROUND: Cavitation of microbubble contrast agents with ultrasound produces shear-mediated vasodilation and an increase in tissue perfusion. We investigated the influence of the size of the cavitation volume by comparing flow augmentation produced by two-dimensional (2D) versus three-dimensional (3D) therapeutic ultrasound. We also hypothesized that cavitation could augment flow beyond the ultrasound field through release of vasodilators that are carried downstream. METHODS: In 11 rhesus macaques, cavitation of intravenously administered lipid-shelled microbubbles was performed in the proximal forearm flexor muscles unilaterally for 10 min. Ultrasound cavitation (1.3 MHz, 1.5 MPa peak negative pressure) was performed with 2D or 3D transmission with beam elevations of 5 and 25 mm, respectively, and pulsing intervals (PIs) sufficient to allow complete postdestruction refill (5 and 12 sec for 2D and 3D, respectively). Contrast ultrasound perfusion imaging was performed before and after cavitation, using multiplane assessment within and beyond the cavitation field in 1.5-cm increments. Cavitation in the hindlimb of mice using 2D ultrasound at a PI of 1 or 5 sec was performed to examine microvascular flow changes from cavitation in only arteries versus the microcirculation. RESULTS: In primates, the degree of muscle flow augmentation in the center of the cavitation field was similar for 2D and 3D conditions (five- to sixfold increase for both, P < .01 vs baseline). The spatial extent of flow augmentation was only modestly greater for 3D cavitation because of an increase in perfusion with 2D transmission that was detected outside of the cavitation field. In mice, cavitation in the microvascular compartment (PI 5 sec) produced the greatest degree of flow augmentation, yet cavitation in the arterial compartment (PI 1 sec) still produced a three- to fourfold increase in flow (P < .001 vs control). The mechanism for flow augmentation beyond the cavitation zone was investigated by in vitro studies that demonstrated cavitation-related release of vasodilators, including adenosine triphosphate and nitric oxide, from erythrocytes and endothelial cells. CONCLUSIONS: Compared with 2D transmission, 3D cavitation of microbubbles generates a similar degree of muscle flow augmentation, possibly because of a trade-off between volume of cavitation and PI, and only modestly increases the spatial extent of flow augmentation because of the ability of cavitation to produce conducted effects beyond the ultrasound field.

Flow Augmentation in the Myocardium by Ultrasound Cavitation of Microbubbles: Role of Shear-Mediated Purinergic Signaling.

BACKGROUND: Ultrasound-mediated cavitation of microbubble contrast agents produces high intravascular shear. We hypothesized that microbubble cavitation increases myocardial microvascular perfusion through shear-dependent purinergic pathways downstream from ATP release that is immediate and sustained through cellular ATP channels such as Pannexin-1. METHODS: Quantitative myocardial contrast echocardiography perfusion imaging and in vivo optical imaging of ATP was performed in wild-type and Pannexin-1-deficient (Panx1-/-) mice before and 5 and 30 minutes after 10 minutes of ultrasound-mediated (1.3 MHz, mechanical index 1.3) myocardial microbubble cavitation. Flow augmentation in a preclinical model closer to humans was evaluated in rhesus macaques undergoing myocardial contrast echocardiography perfusion imaging after high-power cavitation in the apical four-chamber plane for 10 minutes. RESULTS: Microbubble cavitation in wild-type mice (n = 7) increased myocardial perfusion by 64% ± 25% at 5 minutes and 95% ± 55% at 30 minutes compared with baseline (P < .05). In Panx1-/- mice (n = 5), perfusion increased by 28% ± 26% at 5 minutes (P = .04) but returned to baseline at 30 minutes. Myocardial ATP signal in wild-type (n = 7) mice undergoing cavitation compared with sham-treated controls (n = 3) was 450-fold higher at 5 minutes and 90-fold higher at 30 minutes after cavitation (P < .001). The ATP signal in Panx1-/- mice (n = 4) was consistently 10-fold lower than that in wild-type mice and was similar to sham controls at 30 minutes. In macaques (n = 8), myocardial perfusion increased twofold in the cavitation-exposed four-chamber plane, similar in degree to that produced by adenosine, but did not increase in the control two-chamber plane. CONCLUSIONS: Cavitation of microbubbles in the myocardial microcirculation produces an immediate release of ATP, likely from cell microporation, as well as sustained release, which is channel dependent and responsible for persistent flow augmentation. These findings provide mechanistic insight by which cavitation improves perfusion and reduces infarct size in patients with myocardial infarction.

Contrast-enhanced ultrasound detects changes in microvascular blood flow in adults with sickle cell disease.

In patients with sickle cell disease (SCD), poor outcome measures compromise the potential success of clinical trials. Contrast-enhanced ultrasound (CEUS) is a technique that can non-invasively quantify deep tissue microvascular blood flow. We tested the hypothesis that CEUS of forearm skeletal muscle could be used to: 1) assess microvascular abnormalities that occur during vaso-occlusive crisis; and 2) test new therapies for SCD that are targeted to improving the status of the microcirculation. We performed a prospective study, CEUS perfusion imaging of resting forearm muscle was performed in adults with SCD: 1) during and after a pain episode, and 2) before, during, and after a 24-hour infusion of the investigative agent, regadenoson, an adenosine A2A agonist. CEUS destruction-replenishment time-intensity data were analyzed to measure microvascular blood flow, as well as its components, microvascular blood volume and flux rate. Serial CEUS measurements were obtained in 32 adults with SCD. For the studies during crisis, there was a 30% reduction in microvascular blood flow compared to steady-state (p = 0.031), a reduction that was largely due to microvascular flux rate. For the regadenoson group, a non-significant 25% increase in flux rate and 9% increase in microvascular blood flow compared to baseline were detected during infusion. In a study of adults with SCD, CEUS detected changes in microvascular blood flow associated with vaso-occlusive crises. No changes were found during an infusion of the adenosine A2A agonist, regadenoson. This study provides preliminary evidence that CEUS could detect blood flow changes consistent with SCD physiology.

Coronary autoregulation is abnormal in syndrome X: insights using myocardial contrast echocardiography.

BACKGROUND: Syndrome X in women is thought to be caused by coronary microvascular dysfunction, the exact site of which is unknown. The aim of this study was to characterize the microvascular site of dysfunction in these patients using myocardial contrast echocardiography. METHODS: Women with exertional angina, positive test results on stress imaging, but no coronary artery disease (the study group, n = 18) and age-matched control women also with no coronary artery disease (n = 17) were enrolled. Myocardial contrast echocardiography was performed at rest and during dipyridamole-induced hyperemia. Mean microbubble velocity (ß) and myocardial blood volume (A) were measured, and myocardial blood flow (A · ß) was computed. In addition, plasma concentrations of eicosanoids, female sex hormones, and C-reactive protein were measured. RESULTS: Rest ß and myocardial blood flow (A · ß) were higher in the study compared with the control women (1.61 ± 0.68 vs. 0.74 ± 0.44, P = .0001, and 157 ± 121 vs. 54 ± 54, P = 0.0001, respectively) despite similar heart rates and systolic blood pressures. After the administration of dipyridamole, whereas the changes in A and A · ß were not significantly different between the two groups, ß reserve (the ratio of stress ß to rest ß) was markedly lower in the study group (1.48 ± 0.62 vs. 2.78 ± 0.94, P = .0001). Blood hematocrit, eicosanoids, female sex hormones, glucose, and C-reactive protein were not different between the two groups. CONCLUSIONS: Coronary autoregulation is abnormal in patients with syndrome X (higher resting ß and myocardial blood flow and lower ß reserve), which suggests that the coronary resistance vessels are the site of microvascular abnormality.

Molecular imaging of inflammation and platelet adhesion in advanced atherosclerosis effects of antioxidant therapy with NADPH oxidase inhibition.

BACKGROUND: In atherosclerosis, local generation of reactive oxygen species amplifies the inflammatory response and contributes to plaque vulnerability. We used molecular imaging to test whether inhibition of NADPH oxidase with apocynin would reduce endothelial inflammatory activation and endothelial-platelet interactions, thereby interrupting progression to high-risk plaque phenotype. METHODS AND RESULTS: Mice deficient for both the low-density lipoprotein receptor and Apobec-1 were studied at 30 weeks of age and again after 10 weeks with or without apocynin treatment (10 or 50 mg/kg per day orally). In vivo molecular imaging of vascular cell adhesion molecule-1 (VCAM 1) P-selectin, and platelet glycoprotein-1bα (GPIbα) in the thoracic aorta was performed with targeted contrast-enhanced ultrasound molecular imaging. Arterial elastic modulus and pulse wave transit time were assessed using ultrahigh frequency ultrasound and invasive hemodynamic measurements. Plaque size and composition were assessed by histology. Molecular imaging in nontreated mice detected a 2-fold increase in P-selectin expression, VCAM-1 expression, and platelet adhesion between 30 and 40 weeks of age. Apocynin reduced all of these endothelial events in a dose-dependent fashion (25% and 50% reduction in signal at 40 weeks for low- and high-dose apocynin). Apocynin also decreased aortic elastic modulus and increased the pulse transit time. On histology, apocynin reduced total monocyte accumulation in a dose-dependent manner as well as platelet adhesion, although total plaque area was reduced in only the high-dose apocynin treatment group. CONCLUSIONS: Inhibition of NADPH oxidase in advanced atherosclerosis reduces endothelial activation and platelet adhesion, which are likely responsible for the arrest of plaque growth and improvement of vascular mechanical properties.

Ultrasound-mediated vascular gene transfection by cavitation of endothelial-targeted cationic microbubbles.

OBJECTIVES: Ultrasound-mediated gene delivery can be amplified by acoustic disruption of microbubble carriers that undergo cavitation. We hypothesized that endothelial targeting of microbubbles bearing cDNA is feasible and, through optimizing proximity to the vessel wall, increases the efficacy of gene transfection. BACKGROUND: Contrast ultrasound-mediated gene delivery is a promising approach for site-specific gene therapy, although there are concerns with the reproducibility of this technique and the safety when using high-power ultrasound. METHODS: Cationic lipid-shelled decafluorobutane microbubbles bearing a targeting moiety were prepared and compared with nontargeted microbubbles. Microbubble targeting efficiency to endothelial adhesion molecules (P-selectin or intercellular adhesion molecule [ICAM]-1) was tested using in vitro flow chamber studies, intravital microscopy of tumor necrosis factor-alpha (TNF-α)-stimulated murine cremaster muscle, and targeted contrast ultrasound imaging of P-selectin in a model of murine limb ischemia. Ultrasound-mediated transfection of luciferase reporter plasmid charge coupled to microbubbles in the post-ischemic hindlimb muscle was assessed by in vivo optical imaging. RESULTS: Charge coupling of cDNA to the microbubble surface was not influenced by the presence of targeting ligand, and did not alter the cavitation properties of cationic microbubbles. In flow chamber studies, surface conjugation of cDNA did not affect attachment of targeted microbubbles at microvascular shear stresses (0.6 and 1.5 dyne/cm(2)). Attachment in vivo was also not affected by cDNA according to intravital microscopy observations of venular adhesion of ICAM-1-targeted microbubbles and by ultrasound molecular imaging of P-selectin-targeted microbubbles in the post-ischemic hindlimb in mice. Transfection at the site of high acoustic pressures (1.0 and 1.8 MPa) was similar for control and P-selectin-targeted microbubbles but was associated with vascular rupture and hemorrhage. At 0.6 MPa, there were no adverse bioeffects, and transfection was 5-fold greater with P-selectin-targeted microbubbles. CONCLUSIONS: We conclude that ultrasound-mediated transfection at safe acoustic pressures can be markedly augmented by endothelial juxtaposition.

Temporal characterization of the functional density of the vasa vasorum by contrast-enhanced ultrasonography maximum intensity projection imaging.

OBJECTIVES: We sought to determine whether contrast-enhanced ultrasound (CEU) microangiography with maximum intensity projection (MIP) processing could temporally evaluate proliferation of the vasa vasorum (VV) in a model of mural hemorrhage. BACKGROUND: Expansion of the VV and plaque neovascularization contributes to plaque growth and instability and may be triggered by a variety of stimuli, including vascular hemorrhage. However, quantitative in vivo methods for temporal assessment of VV remodeling are lacking. METHODS: In 24 rabbits fed a high-fat diet, either autologous whole blood or saline was percutaneously injected into the media-adventitia of the femoral artery using ultrahigh-frequency ultrasound guidance. Functional VV density at the injection site and contralateral control artery was assessed 1, 2, and 6 weeks after injection with CEU imaging with MIP processing. In vitro studies with renathane microtubes were also performed to validate linear density measurement with CEU and MIP processing. RESULTS: In vitro studies demonstrated that MIP processing of CEU data reflected the relative linear density of vessels in a manner that was relatively independent of contrast concentration or microtube flow rate. On CEU with MIP, there was a 3-fold increase in femoral artery VV microvascular density at 1 and 2 weeks after blood injection (p < 0.01 vs. contralateral control), whereas VV density increased minimally after saline injection. At 6 weeks, VV vascular density decreased in blood-treated vessels and was not different from saline-injected or contralateral control vessels. CONCLUSIONS: CEU with MIP processing can provide quantitative data on temporal changes in the functional density of the VV. This method may be useful for evaluating high-risk features of plaque neovascularization or response to therapies aimed at plaque neovessels.

Molecular imaging of activated von Willebrand factor to detect high-risk atherosclerotic phenotype.

OBJECTIVES: We hypothesized that noninvasive molecular imaging of activated von Willebrand factor (vWF) on the vascular endothelium could be used to detect a high-risk atherosclerotic phenotype. BACKGROUND: Platelet-endothelial interactions have been linked to increased inflammatory activation and prothrombotic state in atherosclerosis. These interactions are mediated, in part, by platelet glycoprotein (GP) Ibα, suggesting that dysregulated endothelial vWF is a marker for high-risk atherosclerotic disease. METHODS: Microbubbles targeted to activated vWF were prepared by surface conjugation of recombinant GPIbα. Flow-chamber studies were used to evaluate attachment of targeted microbubbles to immobile platelet aggregates bearing activated vWF. Contrast-enhanced ultrasound (CEU) molecular imaging of the aorta from mice was performed: 1) ex vivo after focal crush injury and blood perfusion; and 2) in vivo in mice with advanced atherosclerosis produced by deletion of the low-density lipoprotein receptor and ApoBec-1 editing peptide (LDLR(-/-)/ApoBec-1(-/-)). RESULTS: In flow-chamber studies, tracer attachment to vWF was >10-fold greater for microbubbles bearing GPIbα compared with control microbubbles (p < 0.01). In the ex vivo aortic injury model, CEU signal enhancement for vWF-targeted microbubbles occurred primarily at the injury site and was 4-fold greater than at noninjured sites (p < 0.05). In LDLR(-/-)/ApoBec-1(-/-) mice, inflammatory cell infiltrates and dense vWF expression on the intact endothelium were seen in regions of severe plaque formation. Scanning electron microscopy demonstrated widespread platelet-endothelial interaction and only few sites of endothelial erosion. On CEU, signal enhancement for vWF-targeted microbubbles was approximately 4-fold greater (p < 0.05) in LDLR(-/-)/ApoBec-1(-/-) compared with wild-type mice. En face aortic microscopy demonstrated regions where platelet adhesion and microbubble attachment colocalized. CONCLUSIONS: Molecular imaging using GPIbα as a targeting moiety can detect the presence of activated vWF on the vascular endothelium. This strategy may provide a means to noninvasively detect an advanced prothrombotic and inflammatory phenotype in atherosclerotic disease.

A predictive instrument using contrast echocardiography in patients presenting to the emergency department with chest pain and without ST-segment elevation.

OBJECTIVE: Risk stratification of patients presenting to the emergency department (ED) with suspected cardiac chest pain (CP) and an undifferentiated electrocardiogram (ECG) is difficult. We hypothesized that in these patients a risk score incorporating clinical, ECG, and myocardial contrast echocardiography (MCE) variables would accurately predict adverse events occurring within the next 48 hours. METHODS: Patients with CP lasting for 30 minutes or more who did not have ST-segment elevation on the ECG, were enrolled. Regional function (RF) and myocardial perfusion (MP) were assessed by MCE. A risk model was developed in the initial 1166 patients (cohort 1) and validated in subsequent 720 patients (cohort 2). Any abnormality or ST changes on ECG (odds ratio [OR] 2.5; 95% confidence interval [CI], 1.4-4.5, P = .002, and OR 2.9, 95% CI, 1.7-4.8, P < .001, respectively), abnormal RF with normal MP (OR 3.5, 95% CI, 1.8-6.5, P < .001), and abnormal RF with abnormal MP (OR 9.6, 95% CI, 5.8-16.0, P < .001) were found to be significant multivariate predictors of nonfatal myocardial infarction or cardiac death. RESULTS: The estimate of the probability of concordance for the risk model was 0.82 for cohort 1 and 0.83 for cohort 2. The risk score in both cohorts stratified patients into 5 distinct risk groups with event rates ranging from 0.3% to 58%. CONCLUSION: A simple predictive instrument has been developed from clinical, ECG, and MCE findings obtained at the bedside that can accurately predict events occurring within 48 hours in patients presenting to the ED with suspected cardiac CP and an ECG that is not diagnostic for acute ischemic injury. Its application could enhance care of patients with CP in the ED. For instance, patients with a risk score of 0 could be discharged from the ED without further workup. However, this needs to be validated in a multicenter study.

Effect of modest alcohol consumption over 1-2 weeks on the coronary microcirculation of normal subjects.

AIMS: It has been reported that imbibing red wine increases coronary blood flow reserve acutely. In the absence of changes in coronary driving pressure, any increases in coronary blood flow reserve should occur through a decrease in capillary resistance, which in turn is determined by capillary dimensions and whole-blood viscosity. Since alcohol intake is unlikely to acutely change capillary dimensions, we hypothesized that it must increase coronary blood flow reserve by reducing whole-blood viscosity. METHODS AND RESULTS: Forty-five normal subjects were randomly assigned to water (n = 12), vodka (n = 11), white wine (n = 11), and red wine (n = 11). Myocardial blood flow reserve was measured at baseline and after up to 2 weeks of beverage consumption using myocardial contrast echocardiography. In addition, whole-blood viscosity and its principal determinants (haematocrit; erythrocyte deformability, mobility, and charge; plasma fibrinogen; and total serum protein, glucose, and lipids) were also measured. Systolic and diastolic blood pressure and heart rate did not change between the two examinations either at rest or following dipyridamole infusion. Neither did myocardial blood flow reserve nor whole-blood viscosity or any of its determinants. Only high-density lipoprotein-2 increased for all alcohol consumers (12.4 +/- 5.3 vs. 10.9 +/- 4.7, P = 0.007). CONCLUSION: It is concluded that modest alcohol consumption for up to 2 weeks does not increase myocardial blood flow reserve. It also does not alter whole-blood viscosity or any of its principal determinants. Therefore, the beneficial cardiovascular effects of modest alcohol consumption over 1-2 weeks cannot be attributed either to its effect on the coronary microcirculation or haemorheology.

Heterogeneous ventricular sympathetic innervation, altered beta-adrenergic receptor expression, and rhythm instability in mice lacking the p75 neurotrophin receptor.

Sympathetic nerves stimulate cardiac function through the release of norepinephrine and the activation of cardiac beta(1)-adrenergic receptors. The sympathetic innervation of the heart is sculpted during development by chemoattractive factors including nerve growth factor (NGF) and the chemorepulsive factor semaphorin 3a. NGF acts through the TrkA receptor and the p75 neurotrophin receptor (p75(NTR)) in sympathetic neurons. NGF stimulates sympathetic axon extension into the heart through TrkA, but p75(NTR) modulates multiple coreceptors that can either stimulate or inhibit axon outgrowth. In mice lacking p75(NTR), the sympathetic innervation density in target tissues ranges from denervation to hyperinnervation. Recent studies have revealed significant changes in the sympathetic innervation density of p75NTR-deficient (p75(NTR-/-)) atria between early postnatal development and adulthood. We examined the innervation of adult p75(NTR-/-) ventricles and discovered that the subendocardium of the p75(NTR-/-) left ventricle was essentially devoid of sympathetic nerve fibers, whereas the innervation density of the subepicardium was normal. This phenotype is similar to that seen in mice overexpressing semaphorin 3a, and we found that sympathetic axons lacking p75(NTR) are more sensitive to semaphorin 3a in vitro than control neurons. The lack of subendocardial innervation was associated with decreased dP/dt, altered cardiac beta(1)-adrenergic receptor expression and sensitivity, and a significant increase in spontaneous ventricular arrhythmias. The lack of p75(NTR) also resulted in increased tyrosine hydroxylase content in cardiac sympathetic neurons and elevated norepinephrine in the right ventricle, where innervation density was normal.

Effect of acoustic power on in vivo molecular imaging with targeted microbubbles: implications for low-mechanical index real-time imaging.

BACKGROUND: The aim of this study was to evaluate the influence of acoustic power on ultrasound molecular imaging data with targeted microbubbles. METHODS: Imaging was performed with a contrast-specific multipulse method at mechanical indexes (MIs) of 0.18 and 0.97. In vitro imaging was used to measure concentration-intensity relationships and to assess whether damping from microbubble attachment to cultured endothelial cells affected signal enhancement. Power-related differences in signal enhancement were evaluated in vivo by P-selectin-targeted and control microbubble imaging in a murine model of hind-limb ischemia-reperfusion injury. RESULTS: During in vitro experiments, there was minimal acoustic damping from microbubble-cell attachment at either MI. Signal enhancement in the in vitro and in vivo experiments was 2-fold to 3-fold higher for high-MI imaging compared with low-MI imaging, which was due to greater pixel intensity, the detection of a greater number of retained microbubbles, and increased point-spread function. Yet there was a linear relationship between high-MI and low-MI data indicating that the relative degree of enhancement was similar. CONCLUSION: During molecular imaging, high-MI protocols produce more robust targeted signal enhancement than low-MI protocols, although differences in relative enhancement caused by condition or agent are similar.

Echocardiographic particle image velocimetry: a novel technique for quantification of left ventricular blood vorticity pattern.

BACKGROUND: In this study, the functionality of echocardiographic particle imaging velocimetry (E-PIV) was compared with that of digital particle imaging velocimetry (D-PIV) in an in vitro model. In addition, its capability was assessed in the clinical in vivo setting to obtain the ventricular flow pattern in normal subjects, in patients with dilated cardiomyopathy, and in patients with mechanical and bioprosthetic mitral valves. METHODS: A silicon sac simulating the human left ventricle in combination with prosthetic heart valves, controlled by a pulsed-flow duplicator, was used as the in vitro model. Particle-seeded flow images were acquired (1) using a high-speed camera from the mid plane of the sac, illuminated by a laser sheet for D-PIV, and (2) using a Siemens Sequoia system at a frame rate of 60 Hz for E-PIV. Data analysis was performed with PIVview software for D-PIV and Omega Flow software for E-PIV. E-PIV processing was then applied to contrast echocardiographic image sets obtained during left ventricular cavity opacification with a lipid-shelled microbubble agent to assess spatial patterns of intracavitary flow in the clinical setting. RESULTS: The velocity vectors obtained using both the E-PIV and the D-PIV methods compared well for the direction of flow. The streamlines were also found to be similar in the data obtained using both methods. However, because of the superior spatial resolution of D-PIV, some smaller scale details were not revealed by E-PIV. The application of E-PIV to the human heart resulted in reproducible flow patterns in echocardiographic images taken within different time frames or by independent examiners. CONCLUSIONS: The E-PIV technique appears to be capable of evaluating the major flow features in the ventricles. However, the bounded spatial resolution of ultrasound imaging limits the small-scale features of ventricular flow to be revealed.

Effect of time delay on the diagnostic use of contrast echocardiography in patients presenting to the emergency department with chest pain and no S-T segment elevation.

BACKGROUND: The purpose of this study was to assess the effect of time delay on the diagnostic and prognostic use of contrast echocardiography (CE) in patients presenting to the emergency department (ED) with chest pain (CP) and no S-T segment elevation. METHODS: Patients (n = 957, 498 men) presenting to the ED within 12 hours of suggested cardiac CP underwent CE. Regional function (RF) and myocardial perfusion were interpreted separately by expert readers blinded to all other clinical data. Primary (acute myocardial infarction and total mortality) and secondary (unstable angina and revascularization) events within 24 hours of enrollment were determined. RESULTS: Patients were divided into 4 quartiles based on the time interval between their last episode of CP and CE. Patients in quartile I had CE during CP (time delay of 0 minutes). The time delay in quartiles II, III, and IV were 54 +/- 45, 213+/-54, and 556 +/- 184 minutes, respectively (P < .001). The incidence of events was similar among the 4 quartiles. In each quartile, patients with normal RF had the lowest incidence of events, whereas those with both abnormal RF and myocardial perfusion had the highest incidence of events. Patients with abnormal RF but normal myocardial perfusion had an intermediate event rate. CONCLUSIONS: In patients presenting to the ED within 12 hours of CP, the timing of CE does not influence its ability to predict events that occur 24 hours later. These findings have important implications in the performance of CE in the ED.

Automated quantification of the spatial extent of perfusion defects and viability on myocardial contrast echocardiography.

The spatial extent of hypoperfusion or viability is important in the treatment of patients with coronary artery disease. We hypothesized that computerized pixel intensity threshold analysis (PITA) could be used for the automated analysis of perfusion defect size during myocardial contrast echocardiography (MCE). For calibration studies, MCE was performed in 6 dogs undergoing ischemia and reperfusion. Infarct size was determined by PITA, which automatically calculates the percentage of pixels within the myocardium that fail to exceed a predetermined threshold of maximum contrast enhancement. A threshold of 10% of maximum yielded infarct sizes that most closely correlated with those determined by histologic staining. For clinical validation, MCE was performed in 30 patients with acute myocardial infarction before primary percutaneous coronary intervention (PCI) for measurement of risk area; and within 5 days and at 4 weeks after PCI to determine infarct size. The defect size by PITA with a 10% threshold value closely correlated with those measured by expert reader planimetry on background-subtracted color-coded image sets (r = 0.95, P < .001). We conclude that automated analysis of perfusion defect size on MCE is possible by PITA. This technique may be useful for rapid and objective analysis of the extent of ischemia and viability, and for clinical experimentation where accurate and sequential analysis of perfusion defect size is imperative.

Improvement in microvascular reflow and reduction of infarct size with adenosine in patients undergoing primary coronary stenting.

The aim of this study was to use myocardial contrast echocardiography to evaluate the effect of intravenous adenosine on microvascular reflow in patients with acute myocardial infarction who underwent primary coronary stenting (PCS). Thirty patients who underwent primary PCS for acute myocardial infarction were randomized to intravenous adenosine (50 to 70 mug/kg/min) or vehicle for 3 hours. Myocardial contrast echocardiography was performed before and sequentially after PCS to determine the risk area during coronary occlusion and infarct size. The risk area was similar in the adenosine- and placebo-treated patients. The infarct size as a ratio to the risk area was smaller in patients treated with adenosine when measured at 3 to 5 days (0.37 +/- 0.29 vs 0.68 +/- 0.25, p <0.01) and at 4 weeks (0.34 +/- 0.26 vs 0.60 +/- 0.21, p <0.01) after PCS. This effect was greatest when patency was achieved <4 hours after symptom onset (0.18 +/- 0.18 vs 0.74 +/- 0.31, p <0.05), with little effect after 4 hours. The relative microvascular blood volume in the risk area at 4 weeks was higher in patients receiving adenosine than in those receiving placebo (0.73 +/- 0.22 vs 0.57 +/- 0.20, p <0.01), and was highest when patency was achieved in <4 hours. In conclusion, the adjunctive use of intravenous adenosine after PCS reduces the infarct size relative to the risk area. This beneficial effect occurs primarily in those undergoing early intervention.

Myocardial contrast echocardiography versus Thrombolysis In Myocardial Infarction score in patients presenting to the emergency department with chest pain and a nondiagnostic electrocardiogram.

OBJECTIVES: We hypothesized that regional function (RF) and myocardial perfusion (MP) are superior to the Thrombolysis In Myocardial Infarction (TIMI) score for diagnosis and prognostication in patients presenting to the emergency department (ED) with chest pain (CP) and a nondiagnostic electrocardiogram. BACKGROUND: Rapid diagnosis and prognostication is difficult in patients presenting to the ED with suspected cardiac CP. METHODS: Contrast echocardiography was performed to assess RF and MP on 957 patients presenting to the ED with suspected cardiac CP and a nondiagnostic electrocardiogram. A modified TIMI (mTIMI) score was calculated from six immediately available variables. A full TIMI score also was derived after troponin levels were able to be accessed adequately. Follow-up was performed for early (within 24 h), intermediate (30 day), and late primary (death and myocardial infarction) or secondary (unstable angina and revascularization) events. RESULTS: The mTIMI score was unable to discriminate between intermediate- compared to high-risk patients at any follow-up time point, whereas only 2 of 523 patients with normal RF had an early primary event. Regional function provided incremental prognostic value over mTIMI scores for predicting intermediate and late events. In patients with abnormal RF, MP further classified patients into intermediate- and high-risk groups. The full TIMI score could not improve upon these results at any follow-up time point. CONCLUSIONS: Contrast echocardiography can rapidly and accurately provide short-, intermediate-, and long-term prognostic information in patients presenting to the ED with suspected cardiac CP even before serum cardiac markers are known. Integrating contrast echocardiography into the ED evaluation of CP may improve the risk stratification of such patients.