No effects of <i>Plg</i>^T/T mutation in the skin-wound healing model.

<p>The time course of the wound area was measured every other day for two weeks. Data are the means ± SDs of wild-type (WT, n = 10) and <i>Plg</i>^T/T (n = 9) mice. No significant differences (<i>p</i> > 0.05) were observed between groups.</p

Survival time and lung perfusion defect after tissue factor-induced PE.

<p>(<b><i>A</i></b>) After the tissue factor infusion via IVC, the survival time was recorded until 20 min. No significant difference was observed in the survival between wild-type (WT) (n = 28) and <i>Plg</i>^T/T (n = 29) mice (hazard ratio, 1.18; 95% confidence intervals, 0.40–3.50), or between <i>Pros1</i>^E/E (n = 10) and <i>Plg</i>^T/T<i>/Pros1</i>^E/E (n = 10) mice (hazard ratio, 0.81; 95% confidence intervals, 0.27–2.41). The survival rates of both WT and <i>Plg</i>^T/T mice were significantly longer than those of <i>Pros1</i>^E/E and <i>Plg</i>^T/T<i>/Pros1</i>^E/E mice (WT and <i>Pros1</i>^E/E mice: hazard ratio, 5.43; 95% confidence intervals, 1.81–16.28; WT and <i>Plg</i>^T/T<i>/Pros1</i>^E/E mice: hazard ratio, 4.38; 95% confidence intervals, 1.41–13.60; <i>Plg</i>^T/T and <i>Pros1</i>^E/E mice: hazard ratio, 4.62; 95% confidence intervals, 1.61–13.28; <i>Plg</i>^T/T and <i>Plg</i>^T/T<i>/Pros1</i>^E/E mice: hazard ratio, 3.73; 95% confidence intervals, 1.25–11.11). *Significant difference in comparison to WT mice and <i>Plg</i>^T/T mice. (<b><i>B</i></b>) The scale used to measure lung perfusion defect scores. A score of 0 indicates complete perfusion of Evans blue with no occlusion and a score of 4 indicates no Evans blue perfusion with complete occlusion. (<b><i>C</i></b>) Perfusion defect scores. No significant (ns) differences (<i>p</i> > 0.05) were observed among WT (n = 28), <i>Plg</i>^T/T (n = 29), <i>Pros1</i>^E/E (n = 10) and <i>Plg</i>^T/T<i>/Pros1</i>^E/E mice (n = 10). Perfusion defect scores were assessed by the Mann-Whitney test. <i>Circles</i> and <i>diamonds</i> represent individual mouse data. <i>Bars</i> represent the mean values of groups.</p

Thrombus weights in electrolytic IVC injury-induced DVT in wild-type and <i>Plg</i>^T/T mice.

<p>On day 2 or day 7 post-injury, thrombi formed in the IVC were weighed. No significant differences (<i>p</i> > 0.05) were observed in the thrombus weight between wild-type (WT) and <i>Plg</i>^T/T mice (n = 9 and n = 9 on day 2, and n = 5 and n = 5 on day 7, respectively). <i>Circles and squares</i> represent individual mouse data. <i>Bars</i> represent the mean values of groups.</p

Effect of Plg-A622T mutation on fibrinolysis in plasma.

<p>Pooled plasma from 6 wild-type (<b><i>A</i></b>) or 6 <i>Plg</i>^T/T (<b><i>B</i></b>) mice was preincubated in the absence (○) or presence (◆) of human tPA, and clotting was induced with thrombin and CaCl₂. Human α₂-plasmin inhibitor (α₂-PI) was added before the induction of clotting (▽). The turbidity monitored by the absorbance at 405 nm was measured every 5 min as an index of fibrin formation and lysis. The time course of the turbidity was plotted to the maximum absorbance of each sample, which was taken as 100%.</p

No exacerbation of <i>Plg</i>^T/T mutation in the transient focal brain ischaemia model.

<p>(<b><i>A</i></b>) Representative images of coronal sections of wild-type (WT) and <i>Plg</i>^T/T mouse brains. Permanent occlusion of the distal M1 portion of the left middle cerebral artery and 15-min transient occlusion of the bilateral common carotid arteries were applied. After 24 hours, the brains were excised and stained with 2, 3, 5-triphenyl tetrazolium chloride. <i>White</i> areas represent brain infarction. (<b><i>B</i></b>) Infarct volumes. The infarct volume was adjusted for edema by dividing the volume by the edema index (left hemisphere volume / right hemisphere volume). No significant differences (<i>p</i> > 0.05) were observed between groups. <i>Circles</i> represent individual mouse data. <i>Bars</i> represent the mean values of groups.</p

Plasma Plg antigen levels and plasmin activities of wild-type and <i>Plg</i>^T/T mice.

<p>(<b><i>A</i></b>) Plg antigen levels. Data are the means ± SDs of wild-type (WT, n = 10) and <i>Plg</i>^T/T (n = 10) mice. The levels measured in WT mice were defined as 1 U/ml. (<b><i>B</i></b>) Plasmin activities. Plasma from the indicated mice was preincubated with human uPA and reacted with a synthetic substrate, S-2403, for plasmin. Data are the means ± SDs of 10 mice for each genotype. The mean activity measured in WT mice was defined as 100%. (<b><i>C</i></b>) Western blot analysis of uPA-treated mouse plasma. Two mouse plasma samples of each genotype were incubated with human uPA and separated by SDS-PAGE under reducing conditions. Plg (<i>black triangle</i>) and heavy chains of plasmin (<i>white triangles</i>) were detected with anti-mouse Plg antibodies.</p

Generation of Plg-A622T mice.

<p>(<b><i>A</i></b>) Structure of the targeted locus in the mouse <i>Plg</i> gene. Exons are represented by <i>filled boxes</i>. A <i>lox</i>P-flanked (<i>filled triangles</i>) neomycin-resistance cassette (<i>NEO</i>) and a diphtheria toxin A fragment expression cassette (<i>DT-A</i>) are indicated by <i>open boxes</i> with <i>arrows</i> that represent the transcriptional orientation. The A622T mutant allele was produced by homologous recombination and <i>NEO</i> deletion mediated by Cre recombinase. The c.1864G>A (p.A622T) mutation and three translationally silent mutations (c.1857T>G, c.1858C>T, c.1860G>A) creating a new <i>Hpa</i>I site (<i>GTTAAC</i>) were introduced into exon 15. Homologous fragments are indicated by <i>dotted lines</i>, while the <i>Mfe</i>I-<i>Hpa</i>I fragments detected by Southern blot analysis of the wild-type (WT) and Plg-A622T alleles are indicated by <i>double-headed arrows</i>. (<b><i>B</i></b>) Southern blot analysis. Genomic DNA from targeted ES cells was digested with <i>Mfe</i>I/<i>Hpa</i>I and detected with the specific probe (WT allele: 9.5 kb; Plg-A622T allele: 6.6 kb). (<b><i>C</i></b>) Quantitative RT-PCR analysis. Total RNA was extracted from mouse liver and subjected to real-time RT-PCR with dual-labeled probes for mouse <i>Plg</i> and <i>Rn18s</i>. Expression levels of <i>Plg</i> mRNA were normalized to <i>Rn18s</i> mRNA. Data are the means ± SDs of WT (n = 8) and <i>Plg</i>^T/T (n = 7) mice. The levels measured in WT mice were defined as 100%.</p

Additional file 6 of Impact of respiratory bacterial infections on mortality in Japanese patients with COVID-19: a retrospective cohort study

Additional file 6. Admission to intensive care unitand use of invasive mechanical ventilationof bacterial respiratory infection with coronavirus disease 2019

Additional file 1 of Impact of respiratory bacterial infections on mortality in Japanese patients with COVID-19: a retrospective cohort study

Additional file 1. Identification of organisms in ventilator-associated pneumoniacase

Additional file 4 of Impact of respiratory bacterial infections on mortality in Japanese patients with COVID-19: a retrospective cohort study

Additional file 4. Details of respiratory secondary infection