Targeted Knockdown of Hepatic SOAT2 With Antisense Oligonucleotides Stabilizes Atherosclerotic Plaque in ApoB100-only LDLr-/- Mice.

OBJECTIVE: To test the hypothesis that the attenuation of cholesterol oleate packaging into apoB-containing lipoproteins will arrest progression of pre-existing atherosclerotic lesions. APPROACH AND RESULTS: Atherosclerosis was induced in apoB-100 only, LDLr(-/-) mice by feeding a diet enriched in cis-monounsaturated fatty acids for 24 weeks. A subset of mice was then euthanized to quantify the extent of atherosclerosis. The remaining mice were continued on the same diet (controls) or assigned to the following treatments for 16 weeks: (1) a diet enriched in n-3 polyunsaturated fatty acids, (2) the cis-monounsaturated fatty acid diet plus biweekly injections of an antisense oligonucleotide specific to hepatic sterol-O-acyltransferase 2 (SOAT2); or (3) the cis-monounsaturated fatty acid diet and biweekly injections of a nontargeting hepatic antisense oligonucleotide. Extent of atherosclerotic lesions in the aorta was monitored morphometrically in vivo with magnetic resonance imaging and ex vivo histologically and immunochemically. Hepatic knockdown of SOAT2 via antisense oligonucleotide treatment arrested lesion growth and stabilized lesions. CONCLUSIONS: Hepatic knockdown of SOAT2 in apoB100-only, LDLr(-/-) mice resulted in remodeling of aortic atherosclerotic lesions into a stable phenotype, suggesting SOAT2 is a viable target for the treatment of atherosclerosis.

Functionalization of gadolinium metallofullerenes for detecting atherosclerotic plaque lesions by cardiovascular magnetic resonance.

BACKGROUND: The hallmark of atherosclerosis is the accumulation of plaque in vessel walls. This process is initiated when monocytic cells differentiate into macrophage foam cells under conditions with high levels of atherogenic lipoproteins. Vulnerable plaque can dislodge, enter the blood stream, and result in acute myocardial infarction and stroke. Imaging techniques such as cardiovascular magnetic resonance (CMR) provides one strategy to identify patients with plaque accumulation. METHODS: We synthesized an atherosclerotic-targeting contrast agent (ATCA) in which gadolinium (Gd)-containing endohedrals were functionalized and formulated into liposomes with CD36 ligands intercalated into the lipid bilayer. In vitro assays were used to assess the specificity of the ATCA for foam cells. The ability of ATCA to detect atherosclerotic plaque lesions in vivo was assessed using CMR. RESULTS: The ATCA was able to detect scavenger receptor (CD36)-expressing foam cells in vitro and were specifically internalized via the CD36 receptor as determined by focused ion beam/scanning electron microscopy (FIB-SEM) and Western blotting analysis of CD36 receptor-specific signaling pathways. The ATCA exhibited time-dependent accumulation in atherosclerotic plaque lesions of ApoE -/- mice as determined using CMR. No ATCA accumulation was observed in vessels of wild type (C57/b6) controls. Non-targeted control compounds, without the plaque-targeting moieties, were not taken up by foam cells in vitro and did not bind plaque in vivo. Importantly, the ATCA injection was well tolerated, did not demonstrate toxicity in vitro or in vivo, and no accumulation was observed in the major organs. CONCLUSIONS: The ATCA is specifically internalized by CD36 receptors on atherosclerotic plaque providing enhanced visualization of lesions under physiological conditions. These ATCA may provide new tools for physicians to non-invasively detect atherosclerotic disease.

Non-contrasted computed tomography for the accurate measurement of liver steatosis in obese patients.

BACKGROUND: Hepatic macrosteatosis (HMS) is prevalent among high BMI patients, but a lack of validation of non-invasive measures of liver fat hampers non-alcoholic liver disease (NAFLD) investigation in general. Recent work suggests BMI adjusted, non-contrasted computed tomography (nc-CT) attenuation data (Hounsfield units) reflects liver fat accumulation in a normal weight population. However, this and other CT-based HMS studies have only approximated macrosteatosis (%) histologically, but have not validated findings with chemical liver triglyceride (TG) concentrations (mg/gram protein). Also, all previous CT based steatosis studies excluded high BMI subjects, whose habitus may affect properties of the scan. We hypothesized that in high BMI patients nc-CT attenuation measurements expressed in Hounsfield units (HU) accurately estimate liver triglyceride concentrations as well as histological macrosteatosis. METHODS: With informed consent, 15 patients underwent nc-CT scan of the abdomen prior to weight loss surgery with intraoperative wedge and core needle liver biopsy. Mean left lobe nc-CT Hounsfield units (CT(L)), liver TG (mg/g Pr), HMS (%), BMI (kg/m(2)), liver-spleen index (CT(L/S) = hepatic HU/splenic HU), and liver-spleen difference (CT(L-S) = hepatic HU - splenic HU) were a priori outcomes. RESULTS: In 15 patients (11 female) with a BMI of 44.4 ± 1.1 (mean ± SEM), CT(L/S), CT(L-S), and CT(L) measures were significantly associated with liver TG concentrations (r = -0.80, P < 0.001; r = -0.80, P < 0.001; and r = -0.71, P < 0.01, respectively; Table 1). Macrosteatosis (%) and liver triglyceride concentration were positively associated (r = 0.83; P < 0.0001). BMI did not correlate strongly to liver triglyceride (r = 0.44, P = NS). CONCLUSION: Estimates of liver fat obtained by nc- CT scans (esp. CT(L/S), CT(L-S)) correlate to chemical measurement of liver triglyceride concentrations, suggesting non-contrasted CT may be a suitable non-invasive "gold standard" for hepatic steatosis quantification in these patients.

Adverse effect of increased left ventricular wall thickness on five year outcomes of patients with negative dobutamine stress.

BACKGROUND: To determine if patients without dobutamine induced left ventricular wall motion abnormalities (WMA) but an increased LV end-diastolic wall thickness (EDWT) exhibit a favorable cardiac prognosis. RESULTS: Between 1999 and 2001, 175 patients underwent a dobutamine stress cardiovascular magnetic resonance (DCMR) procedure utilizing gradient-echo cines. Participants had a LV ejection fraction >55% without evidence of an inducible WMA during peak dobutamine/atropine stress. After an average of 5.5 years, all participants were contacted and medical records were reviewed to determine the post-DCMR occurrence of cardiac death, myocardial infarction (MI), and unstable angina (USA) or congestive heart failure (CHF) warranting hospitalization.In a multivariate analysis, that took into account Framingham and other risk factors associated with cardiac events, a cine gradient-echo derived LV EDWT > or =12 mm was associated independently with an increase in cardiac death and MI (HR 6.0, p = 0.0016), and the combined end point of MI, cardiac death, and USA or CHF warranting hospitalization (HR 3.0, p = 0.0005). CONCLUSION: Similar to echocardiography, CMR measures of increased LV wall thickness should be considered a risk factor for cardiac events in individuals receiving negative reports of inducible ischemia after dobutamine stress. Additional prognostic studies of the importance of LV wall thickness and mass measured with steady-state free precession techniques are warranted.

Pulmonary extramedullary hematopoiesis.

Extramedullary hematopoiesis (EMH) is the formation and development of blood cells outside of the bone marrow. Of particular interest to chest physicians and radiologists is the occurrence of EMH in the lungs and pleura. There have been several reports of patients presenting with pulmonary EMH published in the literature and the majority are due to a secondary process, such as myeloproliferative disorders, hemolytic anemias, hereditary spherocytosis, and Gaucher disease. We present a case report of pulmonary EMH and a review of the literature concentrating on the etiology, clinical presentation, pathophysiology, diagnosis, and therapy for this uncommon disorder.

Magnetic resonance imaging determination of cardiac prognosis.

BACKGROUND: Regional assessments of left ventricular (LV) wall motion obtained during MRI cardiac stress tests can be used to identify myocardial injury and ischemia, but the utility of MRI stress test results for the assessment of cardiac prognosis is not known. METHODS AND RESULTS: Two hundred seventy-nine patients referred (because of poor LV endocardial visualization with echocardiography) for dobutamine/atropine MRI for the detection of inducible ischemia were followed for an average of 20 months. After MRI stress testing, the occurrence of myocardial infarction, cardiac death, death attributable to any cause, coronary arterial revascularization, and unstable angina or congestive heart failure requiring hospitalization was determined. In a multivariate analysis, the presence of inducible ischemia (hazard ratio 3.3, CI 1.1 to 9.7) or an LV ejection fraction <40% (hazard ratio 4.2, CI 1.3 to 13.9) was associated with future MI or cardiac death independent of the presence of risk factors for coronary arteriosclerosis. CONCLUSIONS: In patients with poor echocardiograms, the results of cardiac MRI stress tests can be used to forecast myocardial infarction or cardiac death.

Effect of the transmural extent of myocardial scar on left ventricular systolic wall thickening during intravenous dobutamine administration.

Using magnetic resonance imaging, the extent of scar tissue due to chronic infarction and quantification of dobutamine systolic wall thickening (SWT) can be measured simultaneously in human subjects. To determine whether the transmural extent of scar tissue determines dobutamine SWT in chronic ischemic heart disease, we assessed the transmural extent of hyperenhancement and dobutamine SWT with magnetic resonance imaging in 16 patients. The transmural extent of hyperenhancement correlated inversely with dobutamine SWT (r = -0.7, p <0.001). All segments with dobutamine SWT >/=2 mm showed a transmural extent of hyperenhancement of <50%.

Relation of cardiac prognosis to segment location with apical left ventricular ischemia.

Previously, we have shown that myocardial ischemia induced during intravenous dobutamine in the distal segments composing the left ventricular apex is associated with a heightened risk of future myocardial infarction or cardiac death. In this study, we find this heightened risk is present independent of the location of the distal segment within the left ventricular apex.

Relation of aortic distensibility determined by magnetic resonance imaging in patients > or =60 years of age to systolic heart failure and exercise capacity.

Aortic stiffness increases with advancing age and is associated with the age-related decline in exercise capacity in healthy persons. Previous studies have suggested that aortic compliance is reduced in heart failure (HF). Older persons with systolic HF can have particularly severe exercise intolerance. However, the relation between increased aortic stiffness and exercise intolerance in elderly patients with systolic HF has not been examined. Therefore, aortic distensibility of the proximal ascending aorta (assessed by magnetic resonance imaging) and exercise tolerance (assessed by maximal exercise ergometry with expired gas analysis) was measured in 28 subjects (10 healthy subjects aged 20 to 30 years, 10 healthy subjects aged > or =60 years, and 8 subjects aged > or =60 years with systolic HF). Compared with healthy older subjects, patients with systolic HF had markedly decreased distensibility of the proximal aorta (0.5 +/- 0.4 vs 2.2 +/- 1.2 10(-3) mm Hg(-1), p <0.002), decreased peak exercise volume of oxygen consumption (VO2) (858 +/- 248 vs 1,436 +/- 344 ml/min, p <0.001), and increased aortic wall thickness (3.6 +/- 0.7 vs 2.9 +/- 0.4 mm, p <0.04). Aortic distensibility was significantly correlated with peak VO2 (r = 0.80, p <0.0001) and remained so even after being adjusted for age and left ventricular (LV) ejection fraction. These data suggest that decreased aortic distensibility may contribute to exercise intolerance in older patients with HF due to LV systolic dysfunction.

Feasibility to detect severe coronary artery stenoses with upright treadmill exercise magnetic resonance imaging.

We performed treadmill exercise magnetic resonance imaging in 27 patients with exertional chest pain who were referred for contrast coronary angiography to determine the feasibility of this method to identify severe coronary artery stenoses. The sensitivity and specificity for detecting >70% coronary artery luminal diameter narrowings on contrast coronary angiography were 79% and 85%, respectively.

Assessment of cardiac function with MR imaging.

A variety of black or white blood imaging techniques are available for assessing global and regional LV and RV function during cardiovascular MR imaging examinations. In addition to providing information about LV function at rest, these techniques provide diagnostic and prognostic information regarding myocardial ischemia and viability during MR imaging stress tests.

Cardiovascular magnetic resonance imaging.

Recent advancements in magnetic resonance imaging hardware and software permit the assessment of cardiovascular structure and function at rest and during exercise or pharmacology-induced cardiac stress. With these developments, knowledge of cardiovascular imaging protocols in the magnetic resonance imaging environment is critical for nursing personnel. The purpose of this article is to review information pertinent to working in a magnetic resonance imaging environment and to describe the requirements of nursing personnel performing cardiovascular magnetic resonance imaging examinations.

Early stages of in situ bladder regeneration in a rodent model.

Surgical removal of approximately 70% of the bladder (subtotal cystectomy [STC]) was used as a model system to gain insight into the normal regenerative process in adult mammals in vivo. Female F344 rats underwent STC, and at 2, 4, and 8 weeks post-STC, bladder regeneration was monitored via microcomputed tomography scans, urodynamic (bladder function studies) pharmacologic studies, and immunohistochemistry. Computed tomography imaging revealed a time-dependent increase in bladder size at 2, 4, and 8 weeks post-STC, which positively correlated with restoration of bladder function. Bladders emptied completely at all time points studied. The maximal contractile response to pharmacological activation and electrical field stimulation increased over time in isolated tissue strips from regenerating bladders, but remained lower at all time points compared with strips from age-matched control bladders. Immunostaining of the bladder wall of STC rats suggested a role for progenitor cells and cellular proliferation in the regenerative response. Immunostaining and the presence of electrical field stimulation-induced contractile responses verified innervation of the regenerated bladder. These initial studies establish the utility of the present model system for studying de novo tissue regeneration in vivo and may provide guidance with respect to optimization of intrinsic regenerative capacity for clinical applications.

Proton magnetic resonance spectroscopy detection of neurotransmitters in dorsomedial medulla correlate with spontaneous baroreceptor reflex function.

Control of heart rate variability via modulation of sympathovagal balance is a key function of nucleus tractus solitarii and the dorsal motor nucleus of the vagus localized in the dorsomedial medulla oblongata. Normal blood pressure regulation involves precise balance of glutamate (Glu)-glutamine-gamma-aminobutyric acid transmitter systems, and angiotensin II modulates these transmitters to produce tonic suppression of reflex function. It is not known, however, whether other brain transmitters/metabolites are indicators of baroreflex function. This study establishes the concept that comprehensive baseline transmitter/metabolite profiles obtained using in vivo (1)H magnetic resonance spectroscopy in rats with well-characterized differences in resting blood pressure and baroreflex function can be used as indices of autonomic balance or baroreflex sensitivity. Transgenic rats with over-expression of renin [m(Ren2)27] or under-expression of glial-angiotensinogen (ASrAogen) were compared with Sprague-Dawley rats. Glu concentration in the dorsal medulla is significantly higher in ASrAogen rats compared with either Sprague-Dawley or (mRen2)27 rats. Glu levels and the ratio of Glu:glutamine correlated positively with indices of higher vagal tone consistent with the importance of these neurotransmitters in baroreflex function. Interestingly, the levels of choline-containing metabolites showed a significant positive correlation with spontaneous baroreflex sensitivity and a negative correlation with sympathetic tone. Thus, we demonstrate the concept that noninvasive assessment of neurochemical biomarkers may be used as an index of baroreflex sensitivity.

Location of arterial stiffening differs in those with impaired fasting glucose versus diabetes: implications for left ventricular hypertrophy from the Multi-Ethnic Study of Atherosclerosis.

OBJECTIVE: To determine whether middle-aged and older individuals with impaired fasting glucose (IFG), but no clinical evidence of cardiovascular disease, exhibit abnormal changes in proximal thoracic aortic stiffness or left ventricular (LV) mass when compared with healthy counterparts. RESEARCH DESIGN AND METHODS: From the Multi-Ethnic Study of Atherosclerosis, 2,240 subjects with normal fasting glucose (NFG), 845 with IFG, and 414 with diabetes, all aged 45 to 85 years and without preexisting coronary artery disease, underwent MRI determinations of total arterial and proximal thoracic aortic stiffness and LV mass. The presence or absence of other factors known to influence arterial stiffness was assessed. RESULTS: After adjustment for clinical factors known to modify arterial stiffness, proximal thoracic aortic stiffness was not increased in those with IFG compared with those with NFG (1.90 +/- 0.05 versus 1.91 +/- 0.04 10(-3) mmHg(-1), respectively, P = 0.83). After accounting for clinical factors known to influence LV mass, LV mass was increased in those with diabetes relative to those with NFG (150.6 +/- 1.4 versus 145.8 +/- 0.81 g, P < 0.0009) but not in those with IFG in comparison with NFG (145.2 +/- 1.03 versus 145.8 +/- 0.81 g, P = 0.56). CONCLUSIONS: Middle-aged and older individuals with the pre-diabetes state of IFG do not exhibit abnormal proximal thoracic distensibility or LV hypertrophy relative to individuals with NFG. For this reason, an opportunity may exist in those with IFG to prevent LV hypertrophy and abnormal aortic stiffness that is observed in middle-aged and older individuals with diabetes.

Prediction of cardiac events in patients with reduced left ventricular ejection fraction with dobutamine cardiovascular magnetic resonance assessment of wall motion score index.

OBJECTIVES: The purpose of this study was to assess the utility of dobutamine cardiovascular magnetic resonance (DCMR) results for predicting cardiac events in individuals with reduced left ventricular ejection fraction (LVEF). BACKGROUND: It is unknown whether DCMR results identify a poor cardiac prognosis when the resting LVEF is moderately to severely reduced. METHODS: Two hundred consecutive patients ages 30 to 88 (average 64) years with an LVEF 40%. CONCLUSIONS: In individuals with mild to moderate reductions in LVEF (40% to 55%), dobutamine-induced increases in WMSI forecast MI and cardiac death to a greater extent than an assessment of resting LVEF. In those with an LVEF <40%, a dobutamine-induced increase in WMSI does not predict MI and cardiac death beyond the assessment of resting LVEF.

Left ventricular spherical remodeling and apical myocardial relaxation: cardiovascular MR imaging measurement of myocardial segments.

PURPOSE: To prospectively evaluate left ventricular (LV) shape and regional relaxation to determine if rapid, early relaxation of the LV is lost with spherical remodeling of the LV. MATERIALS AND METHODS: This HIPAA-compliant study was approved by the institutional review board. All participants gave written informed consent. Cardiovascular magnetic resonance (MR) imaging and transthoracic echocardiography (TTE) were performed in 18 individuals. Each participant was classified into one of three groups according to LV shape and TTE-derived mitral filling parameters. Pairwise comparisons of cardiovascular MR imaging measurements of LV relaxation were made between healthy individuals and those with spherically shaped LVs. RESULTS: The LV regional relaxation rates were determined in a total of 108 basal, middle, and apical myocardial segments in 18 participants (13 women, five men; age range, 35-76 years). Participants with a spherically shaped LV (sphericity index, <1.5) and a mitral inflow velocity E wave/A wave ratio of less than 1.0 exhibited apical thinning velocities that were lower than those of healthy individuals (sphericity index, > or =1.5) (P < .01). The ratio of LV relaxation velocities in the apical versus middle LV segments correlated significantly with sphericity index (R(2) = 0.53; P = .0005). CONCLUSION: LV apical relaxation velocities in participants with LV spherical remodeling (sphericity index, <1.5) were reduced compared with those of healthy individuals (sphericity index, > or =1.5).

Leg flow-mediated arterial dilation in elderly patients with heart failure and normal left ventricular ejection fraction.

BACKGROUND: flow-mediated arterial dilation (FMAD), an indicator of endothelial function, is reduced in patients with heart failure and reduced left ventricular ejection fraction (HFREF). Many elderly patients with heart failure exhibit a normal left ventricular ejection fraction (HFNEF). It is unknown whether FMAD is severely reduced in the elderly with HFNEF. METHODS AND RESULTS: 30 participants >60 yr of age, 11 healthy, 9 with HFNEF, and 10 with HFREF, underwent a cardiovascular magnetic resonance (CMR) assessment of FMAD in the superficial femoral artery followed within 48 h by symptom-limited exercise with expired gas analysis. Elderly patients with HFREF and HFNEF had severely reduced peak oxygen consumption (Vo(2 peak); 12 +/- 2 and 13 +/- 1 ml.kg(-1).min(-1), respectively) vs. their healthy age-matched contemporaries (20 +/- 3 ml.kg(-1).min(-1)). FMAD was 3.8 +/- 1.3% (0.85 +/- 0.22 mm(2)) in patients with HFREF; it was 12.1 +/- 3.6% (3.1 +/- 1.2 mm(2)) and 13.7 +/- 5.9% (3.9 +/- 1.7 mm(2)), respectively, in patients with HFNEF and age-matched healthy older individuals. After adjustment for age and gender, the association of FMAD with Vo(2) was high in healthy and HFREF subjects (P = 0.05 and 0.02, respectively) but less so in HFNEF participants (P = 0.58). CONCLUSIONS: elderly patients with HFNEF do not exhibit marked reduction in leg FMAD. These data suggest that mechanisms other than impaired femoral arterial endothelial function contribute to the severe exercise intolerance experienced by these individuals.