Baseline variables in patient groups according to symptomatology (n = 182).

<p>Clinical symptoms include stroke, TIA or amaurosis fugax ipsilateral to the stenotic internal carotid artery. BMI, body mass index. Numbers are given as percentage (numbers), *mean (SD), or **median (min–max). Cholesterol, HDL, LDL and Triglycerides levels were obtained in respectively 122, 110, 109, and 118 patients.</p

Increased expression of IL-9 and IL-9R in immune cells from patients with coronary atherosclerosis.

<p>mRNA levels of IL-9 and IL-9R were quantified by real-time RT-PCR in CD3⁺ T cells (<b>A</b> and <b>B</b>) and in monocytes (<b>C</b>) from healthy controls (n = 11) and from patients with stable (SAP, n = 11) and unstable (UAP, n = 17) angina. mRNA levels are related to the reference gene 18S (T cells) and β-actin (monocytes) and normalized to levels in healthy controls. For monocyte analyses, only samples from 9 of the controls and 13 of the patients with unstable angina were available. No IL-9R transcripts were detected in patients or controls in these cells. Bars represent mean±SEM. #p<0.05 versus stable angina, *p<0.05 and **p<0.01 versus controls.</p

Association between plasma levels of MMP-7 and adverse outcome in patients with carotid atherosclerosis.

<p>Panel <b>A</b> shows Kaplan–Meier curve with the cumulative incidence of all-cause mortality during the entire study (mean follow-up 3.5 years) according to dichotomized MMP-7 levels (Cut-off median: 1.96 ng/mL). Panel <b>B</b> shows multi-variable analyses of predictors of all-cause mortality (direct entry). CRP and MMP-7 show expressed per SD change.</p

Increased expression of IL-9 and IL-9R in human atherosclerotic carotid plaques.

<p>mRNA levels of IL-9 (<b>A</b>) and IL-9R (<b>B</b>) in patients with asymptomatic (n = 13) and symptomatic (n = 55) carotid stenosis and in non-atherosclerotic vessels obtained from organ donors (common iliac artery, n = 10) were quantified by real-time RT-PCR. No IL-9R transcripts were detected in control samples. Levels of IL-9 and IL-9R expression are related to reference gene β-actin. Data are presented as mean±SEM. ***p<0.0001 versus controls.</p

IL-9 promotes IL-17 release in PBMCs.

<p>PBMCs from healthy controls (Ctr, n = 5) and patients with unstable angina (UAP, n = 5) were stimulated with IL-9 (100 ng/ml) with and without co-stimulation with PHA (20 µg/ml). Panel <b>A</b> shows the absolute release of IL-17 after culturing for 72 hours as assessed by EIA measurements in cell-free supernatants (controls to the left, patients to the right). Panel <b>B</b> shows the percentage change in IL-17 release when adding IL-9 to unstimulated cells from healthy controls (left) and patients (right). Panel <b>C</b> shows the percentage change in IL-17 release when adding IL-9 to PHA-stimulated cells from healthy controls (left) and patients (right). Data are given as mean±SEM. *p<0.05 versus comparative condition without IL-9 (unstimulated and PHA stimulated, respectively). #p<0.05 versus healthy controls.</p

Increased plasma levels of IL-9 in patients with carotid and coronary atherosclerosis.

<p>Panel <b>A</b> shows plasma levels of IL-9 in patients with asymptomatic (n = 56) and symptomatic (n = 88) carotid plaques and in healthy controls (n = 28). Panel <b>B</b> shows plasma levels of IL-9 in patients with STEMI (n = 42) at admission and at different time points after PCI (2, 7 and 60 days). For comparison, levels were also measured in healthy controls (n = 10). Data are presented as mean±SEM. *p<0.05, **p<0.01 and ***p<0.0001 versus controls. #p<0.05 and ##p<0.01 versus levels at admission.</p

FABP4 is co-localized to macrophages within carotid atherosclerotic plaques.

<p>Panel <b>A</b> shows immunostaining of FABP4 in symptomatic carotid atherosclerotic plaques (n = 2) primarily located to macrophage-rich areas. Representative images obtained with 10x and 40x objective. Panel <b>B</b> shows double immunofluorescent staining of FABP4 (green fluorescence), CD68 (macrophages, red fluorescence) and nucleus (DAPI, blue fluorescence) from symptomatic carotid atherosclerotic plaques (n = 2). The lower panel is a merge of the three pictures above. Panel <b>C</b> shows the correlations of mRNA levels in atherosclerotic plaques between FABP4, ADFP and CD68, respectively.</p

Immunostaining of MMP-7 within atherosclerotic and non-atherosclerotic vessels.

<p>Immuhistochemistry of MMP-7 in carotid atherosclerotic plaques (n = 8, symptoms within the recent 2 months) shows strong immunostaining. Representative images obtained with 100× <b>A</b> and 400× magnification (highlighted, with arrows on positive cells). Panel <b>B</b> shows no or weak immunostaining of MMP-7 in non-atherosclerotic carotid artery obtained from autopsies (n = 5). Panel <b>C</b> shows double immunofluorescent staining of MMP-7 (green fluorescence), CD68 (macrophages, red fluorescence) and nucleus (DAPI, blue fluorescence) from carotid atherosclerotic plaques (n = 4). The lower right panel is a merge of the three pictures.</p

The regulation of MMP-7 expression in primary monocytes.

<p>Panel <b>A</b> shows the effect of oxLDL (20 µg/ml), TNFα (5 ng/ml) or a combination thereof, panel <b>B</b> shows the effect of hypoxia with or without co-stimulation with oxLDL (20 mg/ml), TNFα (5 ng/ml) or a combination thereof. The cells were cultured for 48 hours before experimental starts and cell pellets were harvested 18 hours thereafter. mRNA levels of MMP-7 were quantified by real-time RT-PCR in relation to the expression of the endogenous control gene β-actin. Data are mean±SEM (n = 6) and are given in relation to cells that received vehicle (Ctr) or were cultured in normoxic condition. *p<0.05 versus Ctr (panel <b>A</b>). ***p<0.001 versus all other conditions (panel <b>B</b>).</p

High plasma levels of FABP4 are associated with long-term mortality in patients with acute stroke.

<p>Panel <b>A</b> shows ROC curve analysis for the predictive value of FABP4 for all-cause and CV mortality. AUC and 95% CI are given. Panel <b>B</b> shows Kaplan-Meier curves with the cumulative incidence of all-cause and CV mortality during the entire study [median follow-up 4.4 years (interquartile range: 3.7 to 4.9 years)], according to tertiles of FABP4 at admission. Panel <b>C</b> shows the restricted cubic spline analysis of FABP4 in relation to all-cause mortality. Panel <b>D</b> shows multivariate analyses of FABP4 as an independent predictor of mortality in patients with acute stroke. *3^rd tertile vs. lower 2. Panel <b>E</b> shows the increase in hazard ratios (HR) for the prediction of all-cause and CV mortality when combing tertiles of FABP4 and SSS score (inverse for SSS, ie. increasing severity with higher tertiles). Tertile 1 was set as reference and the combination of T3 for both parameters is shown.</p