Dysregulated selectin expression and monocyte recruitment during ischemia-related vascular remodeling in diabetes mellitus.

OBJECTIVE: Diabetes mellitus (DM) is associated with impaired ischemia-related vascular remodeling and also dysregulation of the inflammatory response. We sought to determine whether impaired selectin-mediated monocyte recruitment in ischemic tissues contributes to blunted ischemia-mediated angiogenesis in DM. METHODS AND RESULTS: Contrast-enhanced ultrasound perfusion imaging and molecular imaging of endothelial P-selectin expression in the proximal hindlimb were performed at 1, 3, and 21 days after arterial ligation in wild-type and db/db mice. Ligation reduced muscle blood flow to ≈0.05 mL/minute per gram in both strains. Significant recovery of flow occurred only in wild-type mice (60%-65% of baseline flow). On molecular imaging, baseline P-selectin signal was 4-fold higher in db/db compared with wild-type mice (P<0.01) but increased minimally at day 1 after ischemia, whereas signal increased approximately 10-fold in wild-type mice (P<0.01). Immunohistology of the hindlimb skeletal muscle demonstrated severely reduced monocyte recruitment in db/db mice compared with wild-type mice. Local treatment with monocyte chemotactic protein-1 corrected the deficits in postischemic P-selectin expression and monocyte recruitment in db/db mice and led to greater recovery in blood flow. CONCLUSION: In DM, there is dysregulation of the selectin response to limb ischemia, which leads to impaired monocyte recruitment, which may be mechanistically related to reduced vascular remodeling in limb ischemia.

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.

Role of collateral blood flow in the apparent disparity between the extent of abnormal wall thickening and perfusion defect size during acute myocardial infarction and demand ischemia.

OBJECTIVES: The aim of this study was to test the hypothesis that the apparent disparity between the circumferential extent of abnormal wall thickening (WT) and that of infarct size (IS) at rest or size of ischemic zone (IZ) during demand ischemia (DI) is principally due to the effects of collateral blood flow (CollBF). BACKGROUND: A disparity has been reported between the circumferential extent of abnormal WT and that of IS at rest or IZ size during DI. METHODS: Wall thickening and CollBF were measured in 18 dogs: at 6 h after coronary occlusion (Group 1, n = 6), and during 40 microg x kg x min(-1) of dobutamine in the presence of either one-vessel (Group 2, n = 6) or two-vessel stenosis (Group 3, n = 6). RESULTS: The apparent overestimation of the IS by the circumferential extent of abnormal WT was due to intermediate levels of CollBF in border zones within the risk area that had escaped necrosis. Although reduced, WT in these regions was commensurate with the level of flow. Similarly, during DI, regions within the IZ exhibiting the worst WT in Group 2 and 3 dogs were those not supplied by CollBF. The regions supplied by CollBF had intermediate WT, which was also commensurate with the level of flow. Only in two Group 3 dogs was tethering seen in small, normally perfused regions that were interspersed between two large IZ. Excluding these few tethered regions, data from different myocardial regions (infarcted, ischemic, CollBF dependent, and normal) were described by a single relation: y = 57(1 - e([-0.72(x - 0.06)])) (r = 0.80, p < 0.001). CONCLUSIONS: Myocardial regions at the margins of ischemic territories contribute to the apparent disparity between the circumferential extent of abnormal WT and IS or IZ during DI. In most circumstances, these regions are supplied by collaterals and their WT is commensurate with the degree of myocardial blood flow. The apparent disparity between the circumferential extent of WT and ischemia is rarely due to myocardial tethering, which is seen only in some instances of multi-vessel disease where a small normal region is interspersed between two large IZs.

Detection of noncritical coronary stenosis at rest without recourse to exercise or pharmacological stress.

BACKGROUND: Currently, the detection of noncritical coronary stenoses requires some form of stress. We hypothesized that these stenoses can be detected at rest without recourse to stress by assessing adaptive changes that occur distally in the microcirculation. METHODS AND RESULTS: Phasic changes in myocardial video intensity (VI) were measured at rest with continuous high-mechanical-index (MI) contrast echocardiography in 15 open-chest dogs. Data were acquired at baseline and in the presence of different degrees of noncritical coronary stenosis. In 6 of these dogs, capillary blood volume was also measured at baseline using high-MI intermittent imaging with triggering performed separately at both end diastole and end systole. During continuous high-MI imaging, a significant increase in systolic VI was noted with coronary stenoses that resulted in progressive increases in the systolic/diastolic VI ratio with greater degrees of stenosis (P=0.003), with a mildly quadratic relation noted between the two: y=1.3. 10(-6). x(2)+0.01x+0.32, P<0.001, r=0.76, SEE=0.14. There was no difference in capillary blood volume between end diastole and end systole at baseline. CONCLUSIONS: Capillary blood volume does not change between diastole and systole in vivo. Phasic changes in VI are noted at baseline during high-MI continuous imaging. The systolic component is negligible at baseline but increases with increasing levels of noncritical coronary stenosis because of adaptive changes in the microcirculation distal to the stenosis. Thus, the measurement of phasic changes in myocardial VI has the potential to detect coronary stenosis at rest without recourse to any form of stress.

Myocardial and microcirculatory kinetics of BR14, a novel third-generation intravenous ultrasound contrast agent.

OBJECTIVES: This study sought to investigate the myocardial and microvascular kinetics of BR14, a novel third-generation ultrasound contrast agent. BACKGROUND: BR14 produces persistent myocardial opacification after the administration of a single intravenous bolus when the left ventricular cavity contrast has considerably diminished. The mechanism of this finding is unknown. METHODS: Nine open-chest dogs with non-critical stenosis of a single coronary artery were given intravenous bolus injections of BR14 during coronary hyperemia. Time versus acoustic intensity (AI) plots were generated from the normal and stenosed beds and myocardial blood flow (MBF) was measured with radiolabeled microspheres. Intravital microscopy was performed on an exteriorized cremaster muscle in 11 wild-type mice to study the microvascular kinetics of the agent. RESULTS: At peak contrast enhancement, the ratio between AI in the stenosed and normal bed was 0.44+/-0.23, which was similar to the radiolabeled microsphere-derived MBF ratio between the two beds (0.45 +/-0.20). At 400 s after injection, the AI ratio between the two beds approximated unity (0.99+/-0.07) despite no changes in MBF, indicating redistribution of the agent. The myocardial kinetics of BR14 was best characterized by a modified lagged normal density function. Only about 3% of administered microbubbles were estimated to be retained in the myocardium. Intravital microscopy showed that most of these bubbles were retained only transiently (2 to 3 s) within capillaries. CONCLUSIONS: BR14 demonstrates redistribution because of transient retention within capillaries. Therefore, similar to (201)Tl, it could potentially be used to detect both coronary stenosis and myocardial viability after a single injection during stress.

Microvascular rheology of Definity microbubbles after intra-arterial and intravenous administration.

The microvascular rheology and extent of pulmonary retention of second-generation microbubble ultrasound contrast agents has not previously been well characterized. We assessed the microvascular behavior of Definity, a lipid-shelled microbubble agent containing perfluoropropane gas, using intravital microscopy of either rat spinotrapezius muscle or mouse cremaster muscle. Immediately after intra-arterial injection, which was performed to model pulmonary retention, larger microbubbles (> 5 microm) were entrapped within small arterioles and capillaries. The retention fraction of microbubbles was low (1.2% +/- 0.1%) and entrapment was transient (85% dislodged by 10 minutes), resulting in no adverse hemodynamic effects. Leukocyte or platelet adhesion at the site of entrapment was not seen. After intravenous injection, no microbubble entrapment was observed and the velocities of microbubbles in arterioles, venules, and capillaries correlated well with those of red blood cells. We conclude that after intravenous injection and pulmonary passage, the microvascular rheology of Definity microbubbles is similar to that of red blood cells. Microbubble entrapment within the pulmonary microcirculation after venous injection should be negligible and transient. These findings are important for establishing the safety of this agent.

Measurement of myocardial blood flow velocity reserve with myocardial contrast echocardiography in patients with suspected coronary artery disease: comparison with quantitative gated Technetium 99m sestamibi single photon emission computed tomography.

BACKGROUND: The ability of high and low mechanical index (MI) imaging methods during myocardial contrast echocardiography (MCE) to assess the physiologic significance of coronary stenoses were compared with technetium 99m sestamibi single photon emission computed tomography (SPECT) in patients. METHODS: Intermittent ultraharmonic imaging (high MI) and power modulation angio (low MI) were performed during continuous infusions of the echo-enhancing contrast agent, Optison, at rest and after dipyridamole stress in 39 patients. Technetium 99m sestamibi SPECT was performed simultaneously. Images from the 3 apical windows were divided into 6 walls. Myocardial blood flow (MBF) velocity and MBF velocity reserve were quantified from pulsing interval versus acoustic intensity MCE curves in each wall using postprocessed images. RESULTS: Approximately 25% of the myocardial walls could not be analyzed from MCE because of artifacts. MBF velocity and MBF derived from both MCE methods increased significantly after dipyridamole in healthy patients (n = 143 and 129 walls for high and low MI, respectively), compared with those with either reversible (n = 11 and 10 walls for high and low MI, respectively) or fixed defects (n = 18 and 14 walls for high and low MI, respectively) on SPECT. Consequently, MBF velocity and MBF reserve were significantly greater for patients with normal perfusion. Receiver operator characteristic curves obtained for MBF velocity reserve provided a sensitivity and specificity of 82% and 87%, respectively, for high MI; versus 64% and 96%, respectively, for low MI imaging after uninterpretable images were excluded from analysis. CONCLUSIONS: Both high and low MI MCE imaging techniques can be used to determine the presence of perfusion defects as identified by technetium 99m sestamibi SPECT. Low MI imaging methods have a number of drawbacks that limit its sensitivity compared with high MI techniques.

Direct effects of dobutamine on the coronary microcirculation: comparison with adenosine using myocardial contrast echocardiography.

The direct effects of dobutamine on capillary blood volume (VOL) and blood flow velocity (VEL) are not known. We hypothesized that these would be more similar to that of adenosine because of its effects on the beta(2) receptors on the coronary circulation. A total of 9 open-chest anesthetized dogs were studied after placement of 2 noncritical stenoses at rest and during separate intracoronary administrations of 5 microg/kg(-1)/min(-1) of adenosine and 2 microg/kg(-1)/min(-1) of dobutamine. VOL and VEL were measured using myocardial contrast echocardiography, wall thickening with 2-dimensional echocardiography, and myocardial blood flow (MBF) with radiolabeled microspheres. Dobutamine increased the rate-pressure product significantly, whereas adenosine had no effect on the rate-pressure product. In the normal myocardium, adenosine had no effect on VOL and increases in MBF were all a result of increases in VEL. Dobutamine also caused mostly an increase in VEL and only a 30% increase in VOL indicating modest capillary recruitment. In the bed with stenosis both drugs attenuated increase in MBF by the same amount, which was associated with an attenuation in the increase in VEL secondary to a 15% increase in capillary resistance because of capillary derecruitment. The MBF-wall thickening relation was described for both drugs by the same function: y = 1 - exp(x) with wall thickening being significantly higher for dobutamine compared with adenosine for each level of MBF. We conclude that the increase in MBF in the normal myocardium with intracoronary dobutamine occurs mostly from an increase in VEL rather than from an increase in VOL. In the bed with a noncritical stenosis, the increases in MBF and VEL are similar for both drugs. Similar to intracoronary adenosine, intracoronary dobutamine also caused capillary derecruitment distal to a noncritical coronary stenosis.

Seeking optimal relation between oxygen saturation and hemoglobin concentration in adults with cyanosis from congenital heart disease.

In patients with cyanosis from congenital heart disease, erythropoiesis is governed by many factors that can alter the expected relation between the oxygen saturation (O(2sat)) and hemoglobin concentration. We sought to define the relation between the O(2sat) and hemoglobin in such patients and to predict an ideal hemoglobin concentration for a given O(2sat). Adults with congenital heart defects and cyanosis were studied prospectively using blood tests and exercise testing. Nonoptimal hemoglobin was defined as any evidence of inadequate erythropoiesis (i.e., iron, folate, or vitamin B(12) deficiency, increased erythropoietin, reticulocytosis, or a right-shifted oxygen-hemoglobin curve). For patients without these factors, a linear regression equation of hemoglobin versus O(2sat) was used to predict the optimal hemoglobin for all patients. Of the 65 patients studied, 21 met all the prestudy criteria for an optimal hemoglobin. For all patients, no correlation was found between O(2sat) and hemoglobin (r = -0.22). However, a strong linear correlation was found for those meeting the criteria for optimal hemoglobin (r = -0.865, p <0.001). The optimal hemoglobin regression equation was as follows: predicted hemoglobin = 57.5 - (0.444 × O(2sat)). A negative correlation was found between the hemoglobin difference (predicted minus measured) and exercise duration on cardiopulmonary exercise testing (r = -0.396, p = 0.005) and the 6-minute walk distance (r = -0.468, p <0.001). In conclusion, a strong relation between O(2sat) and hemoglobin concentration can be shown in stable cyanotic patients and used to predict an optimal hemoglobin. This relation might be useful in defining functional anemia in this group.

Cyclic variation in ultrasonic myocardial integrated backscatter is due to phasic changes in the number of patent myocardial microvessels.

OBJECTIVE: We tested the hypothesis that the cyclic variation in ultrasonic myocardial integrated backscatter (IBS) is due to cardiac contraction-induced changes in the number of patent myocardial microvessels. METHODS: We performed experiments in open-chest dogs in which we increased the number of patent myocardial microvessels without changing cardiac contraction. We achieved this either by direct intracoronary administration of adenosine (group 1; n = 10) or by producing a noncritical coronary stenosis (group 2; n = 7). RESULTS: At baseline, IBS was lowest in systole and highest in diastole. This cyclic variation in IBS was closely associated with the phasic changes in myocardial blood volume that were measured with myocardial contrast echocardiography. During adenosine administration, the diastolic IBS increased from -18.8 +/- 6.5 to -17.5 +/- 6.1 dB (P = .002), with an associated increase in the difference between the systolic and diastolic IBS from 3.8 +/- 1.1 to 4.6 +/- 1.0 dB (P = .009). After a noncritical stenosis was produced, diastolic IBS also increased from -26.6 +/- 8.3 to -25.2 +/- 7.3 dB (P = .001), with an associated increase in the difference between the systolic and diastolic IBS from 3.7 +/- 1.2 to 5.0 +/- 1.0 dB (P = .02). No change in IBS was noted in the bed that did not receive adenosine or the bed that had a stenosis. CONCLUSIONS: The variation in IBS during the cardiac cycle is closely associated with the phasic changes in myocardial blood volume seen during cardiac contraction. When the number of patent myocardial arterioles is increased via adenosine or placement of a noncritical stenosis, diastolic IBS increases with a concomitant increase in IBS cyclic variation. These results may have important clinical applications for the noninvasive diagnosis of noncritical coronary stenosis at rest.

Effect of microbubble exposure to ultrasound on quantitation of myocardial perfusion.

BACKGROUND: The purpose of this study was to determine whether acoustic disruption of microbubbles in the left ventricular (LV) cavity prior to entry in the coronary circulation or their gradual destruction in the myocardium during myocardial contrast echocardiography (MCE) affects quantitative parameters of myocardial perfusion. METHODS: MCE was performed in 12 open chest dogs with both intermittent high-power imaging (IHPI) and real-time low-power imaging (RLPI). To assess the affects of microbubble destruction in the LV cavity, MCE parameters of myocardial perfusion were compared for imaging planes that included versus avoided the LV cavity. To assess the effects of gradual disruption of microbubbles in the microcirculation during RLPI, MCE parameters from frames acquired continuously were compared to that from acquiring only end-systolic frames. RESULTS: Destruction of microbubbles in the LV cavity did not alter perfusion data for either form of imaging unless RLPI was performed using long (6-frame) destructive pulse sequences. With RLPI, a gradual decay in microbubble signal occurred during their myocardial transit, the degree of which was related to the acoustic power. Signal decay during microbubble transit resulted in an overestimation of the microvascular blood velocity (beta-value) and an underestimation of the microvascular blood volume (A-value). CONCLUSIONS: MCE parameters of perfusion at low power can be significantly altered by microbubble destruction in the LV cavity and in the myocardial microcirculation during RLPI. Short microbubble destruction pulse sequences and imaging only at end-systole can reduce these effects.