Additional file 1: of Selection bias in clinical stroke trials depending on ability to consent

Full dataset supporting the manuscript. (CSV 60 kb

Blood markers of inflammation and endothelial dysfunction in cardioembolic stroke: systematic review and meta-analysis

<p><b>Context:</b> Various processes including inflammation and endothelial dysfunction have been implicated in the pathogenesis of cardioembolic (CE) strokes.</p> <p><b>Objective:</b> To review the evidence and investigate the association between immune-inflammatory biomarkers and CE strokes versus other stroke subtypes.</p> <p><b>Methods:</b> We systematically reviewed the literature (sources: MEDLINE, web-based register <a href="http://stroke-biomarkers.com" target="_blank">http://stroke-biomarkers.com</a>, reference lists) with quality assessment and meta-analysis of selected articles.</p> <p><b>Results:</b> The most consistent association was found between C-reactive protein (CRP) and CE strokes when compared to other stroke subtypes (standardized mean difference 0.223 (0.116, 0.343); <i>p</i> < 0.001)</p> <p><b>Conclusions:</b> Our findings confirm a possible association between selected inflammatory biomarkers and CE stroke.</p

Leukocyte percentage assessed by flow cytometry of naive, sham or ischemic animals 48 hours after surgery.

<p>(A) Peripheral PMNs percentage: a significant increase was observed in sham and in ischemic animals with respect to the naive group (p<0.05). (B) Lymphocyte percentage. A significant decrease was detected between naive and ischemic animals (p<0.001) but no differences were shown between sham and ischemic animals. (C) Monocyte percentage. (D) Correlation between the percentage of systemic PMNs and the neurological score detected at 48 hours post-ischemia (p<0.05, r = 0.55). Naive (n = 9), sham (n = 13), ischemia (n = 14). *p<0.05, ***p<0.001.</p

Representative images of lung histology after HE stain.

<p>(A) Naive rat lung showing a normal architecture (100x magnification), (b) indicates normal bronchioles. (B) Naive rat lung with alveolar sacs (s), expanded and non-filled alveoli (a), thin septa (arrow), and blood vessels (bv), (400x magnification). (C–F) Lung lesions observed in both sham and ischemic rats (C) Interstitial inflammatory cellular infiltration (100x magnification). (D) Thickened septa (arrow) and inflammatory cellular infiltration (400x magnification). (E) Patchy areas of cellular consolidation (circle), (100x magnification). (F) Acute bronchiolitis (arrow) surrounded by acute pneumonia (400x magnification).</p

On the upper row: cytometry plots of representative (A) naive and (B) ischemic rats with high neurological score. On the lower row: Strategy used for Lymphocyte identification.

<p>(C) Leukocytes distribution considering their granularity (SSC-H) and their size (FSC-H) (D) Leukocyte distribution according to antibody's labelling. FL1-H represents Fit-C and FL2-H, PE. As shown, lymphocytes are CD11b-FITC^- and GrM-PE^- (E) Lymphocytes view after gating them from graph.</p

Bacterial growth 48(A) 200 µL of lung homogenate and (B) lung imprint.

<p>(C) Number of colonies (CFU) found in lung homogenates expressed as log 10. (D) Number of colonies in lung imprints. In the graphs, each dot represents an animal: naive (n = 5), sham (n = 10) and ischemia (n = 6).</p

Leukocyte percentages, bacteriological analysis and measurement of CXCL1 plasma levels of a new set of animals submitted to a tMCAO model (90 min).

<p>Animals were evaluated at 48(A) Peripheral PMNs percentage: a significant increase was observed in ischemic animals with respect to the naive group (p<0.01). (B) Lymphocyte percentage. A significant decrease was detected between naive and ischemic animals (p<0.01) but no differences were shown between sham and ischemic animals. (C) Monocyte percentage. (D) Number of colonies (CFU) found in lung homogenates expressed as log 10. Significant differences were detected when comparing naive to sham groups (p<0.05) and naive to ischemia groups (p<0.01). (E) Number of colonies in lung imprints. Only the comparison between naive and ischemic animals reached significance (p<0.01). (F) CXCL1 plasma levels in naive, sham and ischemic rats. In agreement with peripheral PMNs percentage data, a significant increase was found in ischemic animals with respect to naive ones (p<0.001). In all graphs, naive (n = 7), sham (n = 7) and ischemia (n = 7).</p

Association between the clusters and outcomes.

<p>Association between the clusters and outcomes.</p

Clinical performances of the molecules for discriminating tPA treated (N = 12) vs. ineligible patients (N = 104).

*<p>The AUC of GST-π was significantly better than AUC of DJ-1 (p = 0.016). No significant difference was obtained between GST-π vs. NDKA (p = 0.11) and NDKA vs. DJ-1 (p = 0.14).</p><p>An AUC above 0.80 was estimated as significant (significance 0.05 and power 0.95, made with Power tests/Sample size item from pROC software).</p

Clinical performances of the 3 best molecules for discriminating early (blood sampling between 0 and 3 h after symptoms onset, N = 22) vs. late stroke patients (blood sampling strictly after 3 h, N = 81).

<p>NPV: negative predictive value/PPV: positive predictive value.</p>*<p>The AUC of GST-π was significantly better than AUC of NDKA and DJ-1 (p = 0.034 and 0.020 respectively). No significant difference was obtained between AUC of NDKA and of DJ-1 (p = 0.63).</p><p>An AUC above 0.74 was estimated as significant (significance 0.05 and power 0.95, made with Power tests/Sample size item from pROC software).</p