Richard Bitar

In Vivo 3D High-Spatial-Resolution MR Imaging of Intraplaque Hemorrhage

PURPOSE To apply magnetic resonance (MR) imaging of intraplaque hemorrhage (IPH), as compared with histologic analysis as the reference standard, to detect T1 hyperintense intraplaque signal and to test the hypothesis that T1 hyperintense material represents blood products (methemoglobin). MATERIALS AND METHODS Institutional review board approval and patient informed consent were obtained. Eleven patients undergoing carotid endarterectomy were examined with MR imaging of IPH, and MR images were assessed for T1 hyperintense intraplaque signal. A total of 160 images per patient were available for coregistration with corresponding histologic slices. Because of endarterectomy specimen size and degradation and processing artifacts, only 97 images were coregistered to corresponding histologic slices. A grid that consisted of 16 segments was overlaid on images for correlation of MR images and histologic slices. Only one of 16 segments was chosen randomly per slide and used in the analysis. Agreement between MR images and histologic slices was measured with the Cohen kappa statistic. RESULTS Strong agreement was seen between MR images and histologic slices, with T1-weighted high signal intensity corresponding to hemorrhagic material (kappa = 0.7-0.8). There was a low 2% false-negative rate for the detection of hemorrhage on the basis of T1-weighted hyperintensity (two of 97 measured segments). The results of diagnostic tests for T1 hyperintense detection of hemorrhage were as follows: sensitivity of 100%, specificity of 80%, positive predictive value of 70%, and negative predictive value of 100% for reader 1 and sensitivity of 94%, specificity of 88%, positive predictive value of 78%, and negative predictive value of 97% for reader 2. CONCLUSION With its high spatial resolution, MR imaging of IPH permits detection of plaque hemorrhage location, resulting in strong agreement between imaging and histologic findings.

Granulocytic differentiation of HL-60 cells results in spontaneous apoptosis mediated by increased caspase expression

HL‐60 cells differentiating into neutrophil‐like cells die an apoptotic death in vitro. Susceptibility to apoptosis is associated with decreased Bcl‐2 protein and mRNA expression; however, the effect of differentiation on the expression of pro‐apoptotic caspases is unknown. Spontaneous apoptosis occurred 6 days after retinoic acid treatment. Western blotting showed loss of Bcl‐2 by day 7, and new expression of ICE (caspase 1) and CPP32 (caspase 3) protein by day 2. Northern analysis demonstrated loss of Bcl‐2 mRNA and increases in ICE mRNA by day 2; CPP32 mRNA was unchanged. Differential Bcl‐2 and ICE mRNA expression was also found when granulocytic differentiation was stimulated by DMSO. Differentiated HL‐60 cell lysates exhibited functional ICE proteolytic activity. De novo caspase expression was responsible for the development of spontaneous apoptosis, since specific inhibitors of ICE (YVAD‐CMK) and CPP32 (DEVD‐CHO), inhibited retinoic acid induced spontaneous apoptosis. Functional maturation and susceptibility to apoptosis are both inducible and linked in this granulocyte precursor cell line.

Impaired apoptotic death signaling in inflammatory lung neutrophils is associated with decreased expression of interleukin-1beta converting enzyme family proteases (caspases)

BACKGROUND Fas and tumor necrosis factor receptor 1 (TNFR1) are membrane proteins that signal for apoptotic cell death by downstream activation of proteins of the interleukin-1 beta converting enzyme (ICE) family. Spontaneous apoptosis is delayed in neutrophils activated by transmigration into an inflammatory focus. In this study we evaluated the effects of transmigration on Fas and TNFR1-induced apoptosis and apoptotic gene expression. METHODS Sprague-Dawley rats were killed 4 hours after intratracheal challenge with 500 micrograms lipopolysaccharide (LPS). Neutrophils isolated from the systemic circulation (circulation) or bronchoalveolar lavage fluid (lung) were incubated with or without an agonistic antibody to Fas (clone CH-11, 100 ng/ml) or TNF (10 ng/ml) for 24 hours. Apoptosis and Fas expression were assessed by flow cytometry. Expression of the antiapoptotic protein Bcl-2 and proapoptotic proteins ICE and CPP32 were measured by Western blots. RESULTS Neutrophils transmigrating into the lung in response to LPS showed delayed apoptosis compared with circulating neutrophils and failed to undergo apoptosis in response to anti-Fas antibody or TNF-alpha. Fas expression was unaltered; however, TNFR1 expression was reduced. Bcl-2 was not detected in either group; both the pro- and active forms of ICE and active CPP32 were significantly decreased in lung neutrophils. The specific ICE inhibitor, YVAD-CMK, partially blocked the increased rates of apoptosis resulting from engagement of Fas or TNFR1. CONCLUSIONS Neutrophil transmigration retards apoptosis through engagement of the death receptors Fas and TNFR1. This refractory state is associated with reduced levels of proapoptotic proteins. Blunted responsiveness to physiologic apoptotic stimuli prolongs neutrophil functional survival during acute inflammation and may contribute to the tissue injury associated with acute respiratory distress syndrome.

Late Stage Complicated Atheroma in Low-Grade Stenotic Carotid Disease: MR Imaging Depiction—Prevalence and Risk Factors

PURPOSE To determine if complicated plaque can be found by using magnetic resonance (MR) imaging-depicted intraplaque hemorrhage (IPH), even among symptomatic patients with low-grade (≤50%) carotid stenosis. MATERIALS AND METHODS The institutional ethics review board approved this retrospective study and waived requirements for written informed consent. Symptomatic patients with bilateral 0%-50% carotid stenosis referred for carotid MR imaging were considered. Risk factors (age, sex, hypertension, diabetes, hyperlipidemia, myocardial infarction, atrial fibrillation, smoking, coronary artery disease, and cerebrovascular disease), medications (antihypertensive drugs, diabetes drugs, statins, and aspirin), and the brain side causing symptoms were recorded. MR-depicted IPH prevalence in the carotid arteries ipsilateral and contralateral to the symptomatic side was compared by using the Fisher exact test. Multivariable regression was used to compare the MR-depicted IPH prevalence, while adjusting for risk factors and medications. RESULTS A total of 217 patients (434 carotid arteries) were included. MR-depicted IPH was found in 13% (31 of 233) of carotid arteries ipsilateral and 7% (14 of 201) of arteries contralateral to symptoms (P < .05). Male sex (P < .05) and increasing age (P < .05) were associated with MR-depicted IPH after controlling for risk factors and medications. CONCLUSION Complicated carotid atheroma can be found among symptomatic patients with low-grade (≤50%) stenosis, and this is associated with male sex and increasing age. MR-depicted IPH may be useful to stratify risk for patients with low-grade carotid stenosis.

Intra- and Interobserver Variability in CT Measurements in Oncology

PURPOSE To assess variability of computed tomographic (CT) measurements of lesions of various sizes and margin sharpness in several organs taken by readers with different levels of experience, as would be found in routine clinical practice. MATERIALS AND METHODS In this institutional review board-approved, HIPAA-compliant retrospective study, 17 radiologists with varying levels of experience independently obtained bidimensional orthogonal axial measurements of 80 lymph nodes, 120 pulmonary lesions, and 120 hepatic lesions, categorized by size and margin sharpness. Repeat measurements were performed 2 or more weeks later. Intraclass correlation coefficients and Bland-Altman plots were used to assess intra- and interobserver variability. RESULTS For long- and short-axis measurements, respectively, overall intraobserver agreement rates were 0.957 (95% confidence interval [CI]: 0.947, 0.966) and 0.945 (95% CI: 0.933, 0.955); interobserver agreement rates were 0.954 (95% CI: 0.943, 0.963) and 0.941 (95% CI: 0.929, 0.951). Both intra- and interobserver agreement differed by lesion size, margin sharpness, location, and reader experience. Agreement ranged from 0.847 to 0.886 for lesions 20 mm or larger versus 0.745-0.785 for lesions smaller than 10 mm, 0.961 to 0.975 for smooth margins versus 0.924-0.942 for irregular margins, 0.955 to 0.97 for lung lesions versus 0.884-0.94 for lymph nodes, and 0.95 to 0.97 for attending radiologists versus 0.928-0.945 for fellows. Measurement variability decreased with increasing lesion size; 95% limits of agreement for short-axis measurements were -11.6% to 6.7% for lesions smaller than 10 mm versus -6.2% to 4.7% for lesions 20 mm or larger. CONCLUSION Overall intra- and interobserver variability rates were similar; in clinical practice, serial CT measurements can be safely performed by different radiologists. Smooth margins, larger lesion size, and greater reader experience resulted in a higher consistency of measurements. Depending on lesion size, increases of 4%-6% or greater in long axis and 5%-7% or greater in short axis and decreases of -6% to -10% or greater in long axis and -6% to -12% or greater in short axis at CT can be considered true changes rather than measurement variation, with 95% confidence.

In Vivo Identification of Complicated Upper Thoracic Aorta and Arch Vessel Plaque by MR Direct Thrombus Imaging in Patients Investigated for Cerebrovascular Disease

OBJECTIVE The objective of this article was to assess the feasibility of MR direct thrombus imaging (MRDTI) to evaluate the prevalence and location of complicated upper thoracic aortic and arch vessel plaque in patients referred for evaluation of cerebrovascular disease. SUBJECTS AND METHODS Patients referred for investigation of cerebrovascular disease by MRI were enrolled. Reasons for referral included transient ischemic attack/amaurosis fugax, acute infarct, remote infarct, or asymptomatic carotid disease. Of the 348 patients initially scanned, 17 were excluded from the analysis. The final patient population included 331 patients (199 men, 132 women; mean age, 67.7 years). Patients were scanned using MRDTI, a 3D, T1-weighted, fat-suppressed spoiled gradient echo that exploits the T1 shortening effects of methemoglobin, directly visualizing hemorrhage/thrombus in the vessel wall, thus identifying complicated plaque. Complicated plaque was defined as a high signal within the atherosclerotic plaque at least twice the signal intensity of muscle. RESULTS Forty-three of 331 patients (13%) had complicated upper thoracic aortic atherosclerotic disease, arch vessel atherosclerotic disease, or both. The upper thoracic aorta was involved in 36 of 43 patients (83.7%), and the left subclavian artery was involved in 14 of 43 patients (32.6%). Both the right subclavian artery and the brachiocephalic artery were involved in one of 43 patients (2.3%). Complicated carotid plaque was seen in 25 of 43 patients (58.1%). CONCLUSION MRDTI can be applied in the detection of complicated plaque in the upper thoracic aorta and arch vessels. Complicated plaque was identified in 13% of the patient population. The upper thoracic aorta was the most common site involved. This technique could be useful for the screening of asymptomatic at-risk patients.

Does tumor status influence cancer patients’ satisfaction with the doctor-patient interaction?

The interaction of patients with their doctors impacts the experience of disease at many levels. It is thus important to measure patient satisfaction with such interaction as an outcome of care. Our goal was to investigate whether tumor status influences patient satisfaction with interaction with their doctors. Specifically, we investigated whether patients with no evidence of disease (NED), localized, or metastatic cancers seen in routine follow-up differ in their satisfaction with the oncologist. Outpatients attending clinics at a major cancer center completed a battery of questionnaires, including the Patient Satisfaction with Doctor (PSQ-MD) questionnaire, a 24-item, self-report instrument. It taps two facets of the doctor-patient exchange: perceived support and physician disengagement. Data concerning tumor status and satisfaction were obtained for 569 patients, sampled to include equivalent numbers of women and men with breast, head and neck, gastrointestinal, genitourinary, or lung cancer, or lymphoma. Controlling for age, marital status, annual family income, stressful life events, and employment status, patients with metastatic disease felt somewhat less supported by their physicians (mean=3.26±0.06) than those with localized disease (mean=3.42±0.04) or NED (mean=3.42±0.03), (analysis of covariance, p< 0.05). Physician disengagement did not differ across the groups (means=1.54±0.06, 1.43±0.04, and 1.47±0.03 respectively). These findings were consistent across cancer diagnoses. Patients with metastatic disease may feel less physician support than those with less advanced cancers. Increasing attention to satisfaction of different patient groups can pave the way to improved quality of care.

Reproducibility of semi-automated measurement of carotid stenosis on CTA

PURPOSE To compare the reproducibility of semi-automated vessel analysis software to manual measurement of carotid artery stenosis on computed tomography angiography (CTA). METHODS Two observers separately analyzed 81 carotid artery CTAs using semi-automated vessel analysis software according to a blinded protocol. The software measured the narrowest stenosis in millimeters (mm), distal internal carotid artery (ICA) in mm, and calculated percent stenosis based on NASCET criteria. One observer performed this task twice on each carotid, the second analysis delayed two months in order to mitigate recall bias. Two other observers manually measured the narrowest stenosis in mm, distal ICA in mm, and calculated NASCET percent stenosis in a blinded fashion. Correlation coefficients were calculated for each group comparing the narrowest stenosis in mm, distal ICA in mm, and NASCET percent stenosis. RESULTS The semi-automated vessel analysis software provided excellent intraobserver correlation for narrowest stenosis in mm, distal ICA in mm, and NACSET percent stenosis (Pearson correlation coefficients of 0.985, 0.954, and 0.977 respectively). The semi-automated vessel analysis software provided excellent interobserver correlation (0.925, 0.881, and 0.892 respectively). The interobserver correlation for manual measurement was good (0.595, 0.625, and 0.555 respectively). There was a statistically significant difference in the interobserver correlation between the semi-automated vessel analysis software observers and the manual measurement observers (P < 0.001). CONCLUSION Semi-automated vessel analysis software is a highly reproducible method of quantifying carotid artery stenosis on CTA. In this study, semi-automated vessel analysis software determination of carotid stenosis was shown to be more reproducible than manual measurement.

Antioxidants increase lipopolysaccharide-stimulated TNFα release in murine macrophages : Role for altered TNFα mRNA stability

Through their effects on gene activation, antioxidants have been reported to modulate cellular expression of several proinflammatory cytokines and adhesion molecules, an effect mediated by preventing translocation of the transcription factor nuclear factor-kappa B (NF-kappa B) into the nucleus. In addition, modulation of the intracellular redox state may have profound effects on cell activation and subsequent gene expression distinct from effects on NF-kappa B; these effects may account for the divergent effects of antioxidants on cytokine gene expression in various reports. In the present studies, we evaluated the effect of the antioxidant, pyrrolidine dithiocarbamate (PDTC), on murine and human myeloid cell tumor necrosis factor alpha (TNF alpha) gene and protein expression. PDTC-enhanced LPS-induced TNF alpha secretion in cells derived from a murine macrophage cell line (J774.1), as well as in primary murine peritoneal macrophages by 4-fold. The effect was both stimulus and species dependent, as TNF alpha secretion was attenuated by PDTC in human THP-1 cells and in murine cells stimulated with zymosan. Northern analysis demonstrated that these effects were evident at the level of mRNA expression. Electrophoretic mobility shift assays confirmed the down-regulatory effect of PDTC on human myeloid NF-kappa B activation, whereas in murine cells no such inhibitory effect was evident. Evaluation of TNF alpha mRNA stability in murine cells demonstrated that the potentiating effect of PDTC on TNF alpha mRNA expression was due to an increase in mRNA half-life from 37 to 93 min. Together, these data suggest that the effect of antioxidants on gene expression are both stimulus and species dependent and illustrate a novel mechanism whereby redox manipulation might modulate TNF alpha expression in vivo.