Quantitative analysis The quantitative analysis was performed measuring the most obstructive sample from the 3 sections in each case, and for the extension of atherosclerosis all plaques from the 3 sections were measured, as exemplified in Figure 2. The most severely obstructed vessel segment was measured based on the knowledge that the shortest diameter even in an oblique section of a tube is the same as the diameter in a cross-section at right angles to the longitudinal CX-6258 cost axis, fact

that is referred to be valid also for wall thickness and luminal vessel diameter [37]. Figure 2A shows perpendicular measures, corresponding to

the most obstructive section. A flat shape of the lumen suggests that the segment is collapsed despite of perfusion fixation. Collapsing might have happened during the embedding procedure. The three segments of each case measured around 6 mm length with less than 1 mm thickness. They were embedded in a same paraffin block and it was difficult to maintain them in perpendicular position. Therefore during embedding in a same paraffin block it was difficult to maintain them in perpendicular position and the sections look oblique. An irregular morphology suggesting a bifurcation area as exemplified EPZ015938 molecular weight in section 3, is probably caused by a positive versus absent or negative vessel remodeling induced by atherosclerosis development [38, 39]. In this section, the upper side represents a fat Nutlin-3a plaque in both sides of the vessel, Ergoloid which is associated with a positive vessel remodeling, and the inferior part, a fibrotic plaque with no vessel remodeling. The obstruction was evaluated by perpendicular measures to the vessel long axis, obtaining external diameter, plaque height, luminal diameter,

% luminal obstruction and % fat area in the plaque. The measurements were made only in one plane, across the lowest diameter, in the Masson’s Trichrome and H&E slides, using the Leica – Quantimet 500 Image Analysis System (Cambridge, UK), by obtaining the following variables: a) vessel diameter (distance comprised by the external elastic membrane); b) potential luminal diameter (distance comprised by the internal elastic membrane); c) height of the plaque, and d) luminal diameter. The % luminal obstruction was calculated using the formula: (potential luminal diameter – luminal diameter)/potential luminal diameter × 100). The % lipid content was calculated by measuring the non-stained plaque regions (total plaque area less the fibromuscular area detected by automatic color detection).