Additional file 3: of Sepsis recognition in the emergency department – impact on quality of care and outcome?

Univariate and multivariate regression analysis for impact on missing sepsis diagnoses (table). (PDF 88 kb

Additional file 1: of Sepsis recognition in the emergency department – impact on quality of care and outcome?

Baseline characteristics of all included patients (table). (PDF 169 kb

Additional file 5: of Sepsis recognition in the emergency department – impact on quality of care and outcome?

Death-censored length of hospital stay according to sepsis recognition (Sepsis-3 definitions). Kaplan-Meier curves with log-rank testing showing the length of stay in recognized (n = 17) and unrecognized (n = 32) patients with sepsis. (PDF 117 kb

Thrombelastogram obtained by rotational thromboelastography (ROTEM).

<p>ROTEM analysis is a point-of-care assay that allows the assessment of the complete clotting process. Functional coagulation parameters like clot-formation time (CFT), maximum clot firmness (MCF), and clotting time (CT) are tested in whole blood.</p

Coagulation parameters, ROTEM analysis and blood count.

<p>P-values are from the Wilcoxon signed rank test; aPTT, activated prothrombin time; PT, thromboplastin time; INR, international normalized ratio; TT, thrombin time; CFT, clot-formation time; MCF, maximum clot firmness; CT, coagulation time; WBC, white blood count.</p><p>Coagulation parameters, ROTEM analysis and blood count.</p

Conventional coagulation factors and platelets before and after therapeutic plasma exchange (TPE) with albumin.

<p>After TPE, aPTT (P = 0.0059), fibrinogen (P = 0.002), antithrombin (P = 0.0059) and platelets (P = 0.0039) showed significant changes.</p

Podosome lifetime is altered in Lasp-1 knockdown macrophages.

<p>Primary human macrophages were treated with Lasp-1-specific siRNA (Oligo A or C) or control siRNA. (<b>A</b>) Examples for podosome dynamics included in the analysis. Shown are single podosomes of cells co-transfected with siRNA and Lifeact-TagGFP2. Podosome lifetime is defined as 1) appearance to dissolution, 2) appearance to fission, or 3) fusion to dissolution. (<b>B–D</b>) Measurement of podosome lifetime from cells treated with Lasp-1-specific siRNA, compared to controls. Double asterisks indicate values highly significant different from controls with <i>P</i><0.004 (means+SD, n = 3×4) (B). (C, D) Graph showing detailed podosome lifetime analysis, data from (B), of cells transfected with Lasp-1-specific siRNA (Oligo A (C; purple); Oligo C (D; brown), compared to controls (C, D; grey; (means+SE; n = 3×4;)).</p

Lasp-1 is a component of the podosome ring structure.

<p>(<b>A, B</b>) Confocal micrographs of primary human macrophages transfected with constructs encoding (A) EGFP-Lasp-1 and (B) EYFP-Vinculin (green), respectively, and stained with vinculin- or Lasp-1-specific antibodies (with Alexa488- or Alexa 568-conjugated secondary antibody) for endogenous vinculin or Lasp-1, respectively, and Cy5-conjugated phalloidin for F-actin (blue). White box indicates detail images below. Bars represent 10 µm. (<b>C, D</b>) 3D reconstruction of single podosomes of primary human macrophages. Left panels: xyz mode, right panels: xzy mode for the same podosome. (C) Cells were transfected with EGFP-Lasp-1 and stained with Alexa568-phalloidin for F-actin (podosome core) or vinculin-specific antibody (with Alexa568-labeled secondary antibody) for vinculin (podosome ring structure; red). (D) Both untransfected cells (stained with Alexa488-phalloidin for F-actin; green) and cells transfected with EYFP-vinculin (green) were stained with Lasp-1-specific antibody (with Alexa568-labeled secondary antibody) for endogenous Lasp-1 (red). (<b>E</b>) Confocal micrographs of a rat smooth muscle cell (A7r5), stained with Alexa-594 phalloidin for F-actin (red, upper panel) or zyxin-specific antibody (with Alexa594-labeled secondary antibody) for endogenous zyxin (red, lower panel) and co-stained with Lasp-1-specific antibody (with Alexa488-labeled secondary antibody) for endogenous Lasp-1 (green). Lasp-1 and its binding partner zyxin colocalize at F-actin-rich podosomes (merge) in PDBu-treated A7r5 cells. Bars represent 25 µm.</p

Additional File 1:

Standard Operating Procedure (SOP) (DOCX 16 kb