Concentrations of cadmium in different sections of symptomatic carotid plaques.

<p>The concentrations of cadmium by g of dry and wet weight in the upstream, stenosis, and downstream sections of endarterectomies from patients with symptomatic carotid stenosis (n = 16). Data are geometric mean (SE).</p

Description of a highly stenotic carotid plaque and the division of the endarerectomies.

<p>The figure shows an endarterectomy (an atherosclerotic plaques, excised at operation) and the arrow shows the direction of the blood flow. The endarterectomy samples were prepared in relation to the blood flow direction and the bifurcation between the internal (ICA) and external (ECA) carotid artery. The inner flow divider was defined as the zero point in the vascular lumen, separating the blood flow to ICA and ECA (A) and was localized at the bifurcation (B) minus 1.5 mm [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0121240#pone.0121240.ref023" target="_blank">23</a>]. From that zero point the stenosis part of the plaque was defined as -1.5 to 4.5 mm. The upstream part was < -1.5 mm and the downstream part as >4.5 mm of the endarterectomy. CCA—common carotid artery.</p

Arachidonate 15-Lipoxygenase Enzyme Products Increase Platelet Aggregation and Thrombin Generation

<div><p>Atherosclerotic cardiovascular diseases are the leading causes of morbidity and mortality worldwide. We have previously shown that arachidonate 15-lipoxygenase B (ALOX15B) is highly expressed in atherosclerotic carotid plaques, and elucidation of mechanisms downstream of activated lipoxygenases may be relevant to our understanding of the genesis of atherosclerotic diseases. We examined 120 carotid plaques from patients with symptomatic carotid artery stenosis and showed that the extent of ALOX15B staining was significantly increased in carotid plaques with thrombosis. Impedance aggregometry analyses showed that the ALOX15B enzyme products 15-HETE and 15-HPETE increased platelet aggregation. By using a calibrated automatic thrombin assay, we showed that the ALOX15B products also increased both peak levels of thrombin and the total endogenous thrombin potential. Moreover, platelet aggregation was increased by addition of cell lysates from ischemic human macrophages, whereas platelet aggregation was reduced after knockdown of ALOX15B in human macrophages. Our data show that ALOX15B expression in human carotid plaques is associated with thrombus formation and that enzyme products of ALOX15B increase platelet aggregation and thrombin generation. We therefore propose that activated ALOX15B in macrophages may play a role in the induction of atherothrombotic events by increasing platelet aggregation and thrombin generation.</p></div

Correlation between cadmium levels in blood and atherosclerotic plaques (endarterectomies).

<p>Scatterplot of cadmium concentrations in blood in relation to those in carotid endarterectomies by g of dry and wet weight (n = 35).</p

ALOX15B is present in atherosclerotic plaques. Serial sections of atherosclerotic plaques from the carotid artery of patients with symptomatic carotid artery stenosis (n = 120) were stained with antibodies against ALOX15B and counterstained with Mayer's hematoxylin.

<p>A representative section is shown in (<b>A</b>). (<b>B</b>) ALOX15B staining in plaques with thrombosis (n = 46) and without thrombosis (n = 74). (<b>C</b>) ALOX15B staining in plaques from patients diagnosed with stroke (n = 62) or TIA (n = 31). (<b>D</b>) ALOX15B staining in plaques classified according to AHA classes: III n = 5; IV n = 38; V n = 18; VI n = 59 (AHA class VI indicates complicated lesions with ruptured fibrous cap and/or surface thrombus). Data are mean ± SEM.</p

Platelet aggregation and thrombin formation are increased by ALOX15B enzyme products.

<p>(A, B) Platelet aggregation was measured in blood (from healthy volunteers) in the presence or absence of 10 nmol/L 15-HETE or 15-HPETE or ethanol control. (<b>A</b>) A representative experiment showing platelet aggregation over time. (<b>B</b>) Area under the curve (AUC) of platelet aggregation. Data are mean ± SEM from 4 independent experiments (i.e. 4 blood donors) analyzed in duplicate. (<b>C, D</b>) Thrombin generation was measured in pooled normal plasma in the presence or absence of 10 nmol/L 15-HETE or 15-HPETE or 2% DMSO control. (<b>C</b>) A representative experiment showing thrombin generation curves. (<b>D</b>) The total amount of thrombin activity assessed as the area under curve (i.e. the endogenous thrombin potential). Data are mean ± SEM from 4 independent experiments analyzed in duplicate.</p

Characteristics of the patients with symptomatic carotid plaques in the clinical substudy (n = 37).

<p>Characteristics of the patients with symptomatic carotid plaques in the clinical substudy (n = 37).</p

Additional file 1 of Accelerometer-measured absolute versus relative physical activity intensity: cross-sectional associations with cardiometabolic health in midlife

Additional file 1: Table 1. Characteristics of the study sample, individuals from the Gothenburg site excluded due to missing measurement of fitness and PA, and the entire SCAPIS sample. Mean (standard deviation). Table 2. PLS model details. Number of PLS components was chosen based on cross validation

Patient and plaque characteristics (n = 120).

<p>AHA class, classification of plaques according to American Heart Association.</p

Platelet aggregation is increased by incubation with lysates from human ischemic macrophages.

<p>(A) ALOX15B mRNA expression in primary human monocyte-derived macrophages incubated in control (21% oxygen) and ischemic (1% oxygen) conditions. (<b>B</b>) 15-HETE production in cell lysates from macrophages incubated in control and ischemic conditions. (<b>C</b>) Area under the curve (AUC) of platelet aggregation measured using cell lysates from control and ischemic macrophages. Data are mean ± SEM from 4 independent experiments (i.e. macrophages from 4 blood donors) analyzed in duplicate.</p