Effect of chest radiography on antibiotic use.

<p>Effect of chest radiography on antibiotic use.</p

Demographics of the study population (<i>n</i> = 697).

<p>Demographics of the study population (<i>n</i> = 697).</p

Age-related respiratory rate, from Liu et al. [10].

<p>Age-related respiratory rate, from Liu et al. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096189#pone.0096189-Liu1" target="_blank">[10]</a>.</p

Lung development in late gestation IGF-1R^−/− mice.

<p><b>A–L</b>, Lungs prepared from IGF-1R^+/+ and IGF-1R^−/− embryos at developmental stages E14.5, E17.5 and E19.5. <b>A–F</b>, Ventral view of whole lungs. <b>G–L</b>, Rim of lung lobe. Abbreviations: AL, apical lobe; AzL, azygous lobe; CL, cardiac lobe; DL, diaphragmatic lobes; LL, left lobe. <b>M–X</b>, Lung histology of IGF-1R^+/+ versus IGF-1R^−/− embryos. H&E stained lung sections at developmental stages E14.5 (<b>M</b>–<b>P</b>), E17.5 (<b>Q</b>–<b>T</b>) and E19.5 (<b>U</b>–<b>X</b>), showing that saccular walls are thicker and acinar buds smaller in IGF-1R^−/− embryos as compared with controls of the same stage. Note that histomorphological appearance is similar when comparing E19.5 IGF-1R^−/− (<b>V</b>, <b>X</b>) with two days younger E17.5 IGF-1R^+/+ lungs (<b>Q</b>, <b>S</b>).</p

Development of diaphragm and chest in the absence of IGF-1R.

<p><b>A</b>, Hematoxylin-eosin stained transversal section of thoracic wall and diaphragm in control (left) and IGF-1R^−/− embryos (right) at E17.5. Bar graphs compare <b>B</b>, diaphragm thickness, <b>C</b>, rib diameter, and <b>D</b>, diaphragm-to-rib ratio (mean ± SEM) in IGF-1R^+/+ embryos (n = 4) and IGF-1R^−/− embryos (n = 4). R, Rib; D, diaphragm. Wilcoxon Mann-Whitney U test.</p

Lung, heart, liver and kidney weight relative to body weight, in IGF-1R^+/+ and IGF-1R^−/− embryos.

<p>Values are mean ± SEM. * <i>P</i><0.05; ** <i>P</i><0.01; *** <i>P</i><0.001 compared with IGF-1R^+/+ mice. Student’s <i>t</i>-test.</p>a<p>n = 5; BW, body weight; ND, not determined.</p

Lung histomorphology and cell turnover in the absence of IGF-1R.

<p><b>A</b>, Saccular airspace and <b>B</b>, saccular wall thickness (mean ± SEM) at developmental stages E17.5 in IGF-1R^+/+ (n = 4) and IGF-1R^−/− embryos (n = 4). Wilcoxon Mann-Whitney U test. <b>C–F,</b> Extended gestation period and lung histology. H&E stain of lung tissue from embryos at 19.5 (<b>C</b>, <b>D</b>) and 21.5 days (<b>E</b>, <b>F</b>). To extend gestation period up to 21.5 days, pregnant mothers were treated with progesterone from E17.5 onwards. Note the presence of red blood cell extravasation in E21.5 lung samples from IGF-1R^+/+ and IGF-1R^−/− mice. <b>G–O,</b> Cell turnover in IGF-1R^−/− embryonic lung at E17.5. Lung histology from IGF-1R^+/+ embryos (G, J and M) and IGF-1R^−/− embryos (H, K and N) at E17.5. Bar graphs (I, L and O) show quantification (mean ± SEM; n = 3–7 individuals per group; Student’s <i>t</i>-test). <b>G–I</b>, Cells were counted using DAPI staining (blue signal). <b>J–L</b>, Cell proliferation was measured using phospho–histone H3 immunohistochemistry (brown staining). <b>M–O</b>, Apoptosis was detected using cleaved caspase-3 immunohistochemistry (brown staining). Cleaved caspase-3 (P-U) and phospho-histone H3 labeling (V-AA) at high magnification showing examples for IHC-positive epithelial (red arrows), vascular endothelial (blue) and mesenchymal cells (green), as identified by their anatomical location. Note that many of the proliferating cells are located in areas that are composed of mostly mesenchymal cells.</p

IGF-1R protein levels, lung histology and respiratory function in adult IGF-1R^neo/− mice. A

<p>, Western immunoblot of IGF-1R in lung from mice with distinct combinations of mutant IGF-1R alleles. Total proteins were extracted from lung tissue from IGF-1R^neo/−, IGF-1R^+/−, IGF-1R^+/neo and IGF-1R^+/+ mice (n = 3 for each genotype), and IGF-1R^−/− embryo (negative control), and were probed with anti-IGF-1Rβ (upper panel) or anti-β-actin antibodies (lower panel). IGF-1R^neo/− mice have 22% of receptor levels present in IGF-1R^+/+ mice (quantified in B), IGF-1R^+/− have 50%, IGF-1R^+/neo mice are between 70 and 80%, and IGF-1R^−/− mice lack IGF-1R completely. <b>B</b>, IGF-1R abundance determined in lung tissue. Bar graph shows IGF-1R levels relative to β-actin from 5 IGF-1R^+/+ and 6 IGF-1R^neo/− individuals (Error bars SEM; Student’s <i>t</i>-test). Image shows 4 representative lanes from western immunoblot. <b>C</b>, Hematoxylin-eosin stained lung sections from IGF-1R^+/+ and IGF-1R^neo/− males. <b>D</b>, Alveolar airspace, <b>E</b>, alveolar boundary length density, <b>F</b>, alveolar wall thickness, in IGF-1R^+/+ (n = 4) and IGF-1R^neo/− mice (n = 4). Error bars indicate SEM; Wilcoxon Mann-Whitney U test. <b>G-I</b>, Respiratory function in adult IGF-1R^neo/− mice. Mice were challenged with 6% and 8% CO₂. <b>G</b>, Minute ventilation (V_E), <b>H</b>, tidal volume (V_T), and <b>I</b>, respiratory frequency (BR) were measured in 6 individuals per group. Differences between room air and hypercapnia were significant, but no significant differences were found between genotypes. Values labeled <i>b</i> were different from <i>a</i> (<i>P</i><0.005); Error bars indicate SEM; Wilcoxon Mann-Whitney U test.</p

Immunohistochemistry of lung differentiation markers.

<p><b>A</b> and <b>B,</b> Representative tissue sections from IGF-1R^+/+ and IGF-1R^−/− embryos at stage E17.5 showing CD31-immunoreactivity specific for capillary endothelia. <b>C</b>, Morphometric comparison of CD31 signal between genotypes (n = 5 per group; two-tailed <i>t</i>-test). <b>D–F</b>, Capillary complexity was estimated calculating the density of capillary junctions from CD31 IHC. <b>G–J</b>, Sections from IGF-1R^+/+ and IGF-1R^−/− embryos at E17.5 and E19.5 show IHC of blood vessel-specific von Willebrand protein. Arrows (I, J) point to small blood vessels developing in saccular walls. Large blood vessels were similarly marked in all specimen. <b>K–M</b>, Representative lung histology from IGF-1R^+/+ and IGF-1R^−/− embryos at E17.5. NKX2-1 distal-to-proximal IHC signal ratio was measured in 6 IGF-1R^+/+ and 5 IGF-1R^−/− embryos. NS, not significant; Wilcoxon Mann-Whitney U test. <b>N-Q</b>, Epithelial cell-specific NKX2-1 transcription factor was detected in IGF-1R^+/+ and IGF-1R^−/− embryos at E17.5 and E19.5. <b>R–Y</b>, IHC of type 2-specific pro-SP-C at low (R-U) and high magnification (V-Y). Interestingly, for NKX2-1 and pro-SP-C, the IHC pattern of IGF-1R^−/− lungs at E19.5 resembles controls at E17.5 (panel N <i>versus</i> Q, R <i>versus</i> U, and V <i>versus</i> Y), suggesting an approximately 2-day developmental delay in IGF-1R^−/− end-gestational lungs.</p

Additional file 1: of Pulmonary hemosiderosis in children with Down syndrome: a national experience

Table S1. Detailed characteristics of the 34 included patients. (DOCX 24 kb