A quantitative neuropathological assessment of translocator protein expression in multiple sclerosis.

The 18 kDa translocator protein (TSPO) is increasingly used to study brain and spinal cord inflammation in degenerative diseases of the CNS such as multiple sclerosis. The enhanced TSPO PET signal that arises during disease is widely considered to reflect activated pathogenic microglia, although quantitative neuropathological data to support this interpretation have not been available. With the increasing interest in the role of chronic microglial activation in multiple sclerosis, characterising the cellular neuropathology associated with TSPO expression is of clear importance for understanding the cellular and pathological processes on which TSPO PET imaging is reporting. Here we have studied the cellular expression of TSPO and specific binding of two TSPO targeting radioligands (3H-PK11195 and 3H-PBR28) in tissue sections from 42 multiple sclerosis cases and 12 age-matched controls. Markers of homeostatic and reactive microglia, astrocytes, and lymphocytes were used to investigate the phenotypes of cells expressing TSPO. There was an approximate 20-fold increase in cells double positive for TSPO and HLA-DR in active lesions and in the rim of chronic active lesion, relative to normal appearing white matter. TSPO was uniformly expressed across myeloid cells irrespective of their phenotype, rather than being preferentially associated with pro-inflammatory microglia or macrophages. TSPO+ astrocytes were increased up to 7-fold compared to normal-appearing white matter across all lesion subtypes and accounted for 25% of the TSPO+ cells in these lesions. To relate TSPO protein expression to ligand binding, specific binding of the TSPO ligands 3H-PK11195 and 3H-PBR28 was determined in the same lesions. TSPO radioligand binding was increased up to seven times for 3H-PBR28 and up to two times for 3H-PK11195 in active lesions and the centre of chronic active lesions and a strong correlation was found between the radioligand binding signal for both tracers and the number of TSPO+ cells across all of the tissues examined. In summary, in multiple sclerosis, TSPO expression arises from microglia of different phenotypes, rather than being restricted to microglia which express classical pro-inflammatory markers. While the majority of cells expressing TSPO in active lesions or chronic active rims are microglia/macrophages, our findings also emphasize the significant contribution of activated astrocytes, as well as smaller contributions from endothelial cells. These observations establish a quantitative framework for interpretation of TSPO in multiple sclerosis and highlight the need for neuropathological characterization of TSPO expression for the interpretation of TSPO PET in other neurodegenerative disorders.

Using team-based revision to prepare medical students for the prescribing safety assessment.

BACKGROUND: The Prescribing Safety Assessment (PSA) is an online assessment of safe and effective prescribing, taken by final-year UK medical students. To prepare students for the PSA, we used a modified form of team-based learning, team-based revision (TBR), in which students consolidate previously learned prescribing knowledge and skills across a broad range of topics. We evaluated students' response to TBR and their perceptions of team working. METHODS: Eight TBR sessions based on the PSA blueprint were conducted over two days by three faculty members for final year medical students. During TBR sessions, students worked in small groups answering individual multiple-choice questions, followed by group multiple-choice questions. They subsequently answered open-ended questions in their groups, with answers written on a drug chart to increase authenticity. Students completed surveys using Likert-type items to determine views on TBR and their confidence in prescribing. RESULTS: The majority of respondents agreed that the sessions were useful for preparation both for the PSA (82%) and Foundation Year 1 (78%). 92% agreed that using drug-charts aided learning. Prescribing confidence increased significantly after TBR (median pre-TBR: 2, post-TBR: 5, p<0.0001). TBR significantly improved attitudes towards "team experience" (p<0.001), "team impact on quality of learning" (p<0.01) and "team impact on clinical reasoning ability" (p<0.001). CONCLUSIONS: Team-based revision is a resource-efficient addition to undergraduate prescribing teaching and can help with preparation for the PSA. A short course of TBR was effective in influencing students' attitudes towards teamwork.

In Vivo Assessment of Brain White Matter Inflammation in Multiple Sclerosis with (18)F-PBR111 PET.

UNLABELLED: PET radioligand binding to the 18-kD translocator protein (TSPO) in the brains of patients with multiple sclerosis (MS) primarily reflects activated microglia and macrophages. We previously developed genetic stratification for accurate quantitative estimation of TSPO using second-generation PET radioligands. In this study, we used (18)F-PBR111 PET and MR imaging to measure relative binding in the lesional, perilesional, and surrounding normal-appearing white matter of MS patients, as an index of the innate immune response. METHODS: (18)F-PBR111 binding was quantified in 11 MS patients and 11 age-matched healthy volunteers, stratified according to the rs6971 TSPO gene polymorphism. Fluid-attenuated inversion recovery and magnetization transfer ratio (MTR) MR imaging were used to segment the white matter in MS patients as lesions, perilesional volumes, nonlesional white matter with reduced MTR, and nonlesional white matter with normal MTR. RESULTS: (18)F-PBR111 binding was higher in the white matter lesions and perilesional volumes of MS patients than in white matter of healthy controls (P < 0.05). Although there was substantial heterogeneity in binding between different lesions, a within-subject analysis showed higher (18)F-PBR111 binding in MS lesions (P < 0.05) and in perilesional (P < 0.05) and nonlesional white matter with reduced MTR (P < 0.005) than in nonlesional white matter with a normal MTR. A positive correlation was observed between the mean (18)F-PBR111 volume of distribution increase in lesions relative to nonlesional white matter with a normal MTR and the MS severity score (Spearman ρ = 0.62, P < 0.05). CONCLUSION: This study demonstrates that quantitative TSPO PET with a second-generation radioligand can be used to characterize innate immune responses in MS in vivo and provides further evidence supporting an association between the white matter TSPO PET signal in lesions and disease severity. Our approach is practical for extension to studies of the role of the innate immune response in MS for differentiation of antiinflammatory effects of new medicines and their longer term impact on clinical outcome.

Assessment of global liver blood flow with quantitative dynamic contrast-enhanced ultrasound.

OBJECTIVES: This study assessed the potential of quantitative analysis of contrast bolus kinetics to reflect global liver blood flow. METHODS: A dynamic contrast-enhanced ultrasound flow phantom was developed. A peristaltic pump established constant volume flow ranging between 16.5 and 49.5 mL/min (2-mm tube) and 85.5 and 256.5 mL/min (5-mm tube). After bolus injection of 2 doses of a contrast agent, a region of interest was drawn over the cross section of the tube used for a particular acquisition; the rise time, peak intensity, and wash-in slope were derived from time-intensity curves. Twenty healthy volunteers and 25 patients with biopsy-proven colorectal liver metastases were scanned with dynamic contrast-enhanced ultrasound. The rise time, peak intensity, and wash-in slope were derived from hepatic artery and portal vein time-intensity curves. Hepatic artery/portal vein ratios of the parameters were also calculated. RESULTS: In the in vitro experiment, the rise time decreased while the peak intensity and wash-in slope increased with increasing volume flow for both tube diameters and contrast bolus volumes. In the clinical study, the rise time was lowered in the hepatic artery but elevated in the portal vein, and the peak intensity and wash-in slope were elevated in the hepatic artery but lowered in the portal vein in patients with colorectal liver metastases compared with healthy volunteers, although not in a statistically significant manner. This finding was consistent with an increase in hepatic artery blood flow, a decrease in portal vein blood flow, or both in patients with colorectal liver metastases compared with healthy volunteers. Only the 3 hepatic artery/portal vein ratios of the parameters achieved statistical significance in differentiating healthy volunteers from patients with colorectal liver metastases (P < .05). CONCLUSIONS: Surrogate measurements of liver blood flow may be derived from quantitative analysis of dynamic contrast-enhanced ultrasound studies. They may have potential for quick and easy assessment of altered hepatic hemodynamics.

Inflammation within carotid atherosclerotic plaque: assessment with late-phase contrast-enhanced US.

PURPOSE: To determine if the number of nontargeted microbubbles retained in human carotid plaque is sufficient to be detected with ultrasonography (US). MATERIALS AND METHODS: The study protocol was approved by the local research ethics committee. Informed consent was obtained. A total of 37 subjects with carotid atherosclerosis (mean age, 69.9 years; age range, 49-86 years), of whom 27 (73%) were men (mean age, 69.7 years; age range, 58-86 years) and 10 (27%) were women (mean age, 70.3 years; age range, 49-86 years), were studied between December 2008 and May 2009 with late-phase (LP) contrast material-enhanced US by using flash imaging with a nonlinear mode at an intermediate mechanical index of 0.34 6 minutes after bolus contrast agent injection. Plaques were defined as symptomatic if symptoms consistent with stroke, transient ischemic attack, or amaurosis fugax had occurred in the neurovascular territory of the plaque studied within 12 months prior to entry into the study. Plaques were defined as asymptomatic if no such events had ever occurred within the neurovascular territory. Raw linear data were used to quantify echogenicity of the plaque, which was normalized to lumen echogenicity. Gray-scale median score was also calculated. RESULTS: Of the 37 subjects, 16 (43%) had symptomatic plaques and 21 (57%) had asymptomatic plaques. All examinations yielded evaluable LP contrast-enhanced US data. Normalized LP plaque echogenicity was greater in the symptomatic group (0.39; 95% confidence interval: -0.11, 0.89) than in the asymptomatic group (0.69; 95% confidence interval: -1.04, -0.34) (P = .0005). There was a moderate (rho = -0.44, P = .016) inverse correlation between normalized LP plaque echogenicity and gray-scale median score. CONCLUSION: By quantifying microbubble retention within the carotid plaque, LP contrast-enhanced US depicts clear differences between groups of subjects with plaque ipsilateral to symptoms and asymptomatic plaques. This technique has promise as a tissue-specific marker of inflammation and a potential role in the risk stratification of atherosclerotic carotid stenosis.

Identification and assessment of plasma lysozyme as a putative biomarker of atherosclerosis.

OBJECTIVE: To identify a plasma biomarker of atheromatous disease. METHODS AND RESULTS: Surface-enhanced laser desorption ionization-time-of-flight mass spectrometry was used to identify possible plasma protein biomarkers of atheromatous disease in patients presenting with chronic stable angina pectoris by comparing those with 3-vessel disease with those without any evidence of coronary artery disease. The level of a 14.7-kDa protein was elevated; this protein was isolated and identified as a lysozyme. Arterial plasma lysozyme levels, measured by immunoassay, confirmed this observation in separate cohorts of patients. The application of arterial plasma lysozyme levels to 197 patients with varying degrees of coronary artery disease, using a cutoff value of 1.5 microg/mL, was able to distinguish patients with 1 or more occluded coronary arteries, with 86% sensitivity and 93% specificity. Of 20 patients with carotid atheroma, 19 had increased arterial plasma levels. In contrast, C-reactive protein levels showed no association with disease severity. Venous lysozyme levels in patients with carotid atheroma were shown to decrease after intensive atorvastatin treatment. CONCLUSION: Raised plasma lysozyme levels may be a useful biomarker of atherosclerotic cardiovascular disease and response to therapy. Additional studies to investigate this are warranted.

The incremental variance problem: enhancing the predictability of academic success in an urban, commuter institution.

The authors identified variables that enhance the predictability of academic performance and retention in an urban, commuter college. They used a longitudinal design with 2 waves of data collection: prior to 1st-semester attendance and again 6 semesters later. The results support the following conclusions: (a) After controlling for precollege indices of academic ability, specific variables within demographic, personal behavior, expectations, and attitudinal domains improved the predictability of grade point average by 15.5% for Whites and 20.3% for Blacks; overall, these analyses accounted for 61% of the variance in grade point average for Whites and 47% for Blacks; (b) for retention, the comparable increment in predictability was 2.5% for Whites and less than 1% for Blacks; overall, the percentage of retention variance accounted for was 42% for Whites and 29% for Blacks; and (c) a paradoxical finding was the combination of a positive correlation between high school average and students' expected grades on tests and a negative correlation between expected and actual grades. The paradox is resolved by citing data that demonstrate (a) a disparity between high school grades and independently measured academic accomplishment; (b) the consequent false attributions students draw about their academic abilities; and (c) the impact of these false attributions on the priority students place on their academic responsibilities.