The presence of emphysema, mucus production and fibrosis in mice with repeated intranasal administration of α-GalCer.

<p>Mice were intranasally administered with α-GalCer once a week for 6 weeks and histopathological changes were examined 2 weeks after the last administration. (A) The mean linear intercept of the alveolar septa was measured. (B) The upper panels show the western blot results of 54 kDa and 45 kDa MMP12 protein expression in the lung and summarized in the lower panels. Actin was used as an internal control. Each symbol represents an individual mouse and the horizontal lines represent the means. (C) The expression of <i>Mmp-12</i> in lung tissues was analyzed by quantitative RT-PCR. (D) The expression of <i>Mmp-12</i> in lung macrophages was measured by quantitative RT-PCR. Each symbol represents cells pooled from 2–3 mice and the horizontal lines represent the means of three symbols. (E) Representative sections of the lung stained with PAS for analysis of mucus-containing cells. (200x magnification). (F) The expression of <i>Muc5ac</i> in lung tissues was analyzed by quantitative RT-PCR. (G) Representative results of Massion’s trichrome staining of lung sections (200x magnification, blue color). (H) The expression of <i>collagen III</i> in lung tissues was evaluated by quantitative RT-PCR. n = 9–14 mice for each group. a-GC, α-GalCer. **, p<0.01; ***, p<0.001 using Mann-Whitney U test.</p

Intranasal α-GalCer administration activates lung iNKT cells to secrete cytokines.

<p>Mice were intranasally administered with vehicle or α-GalCer. Two hours later, BALF was collected. (A) The absolute numbers of iNKT (CD3⁺PBS57 loaded CD1d tetramer⁺) cells were measured. (B) IFN-γ and IL-4 expression from iNKT cells was assayed by intracellular staining with anti-mouse IFN-γ and IL-4 Abs. The shaded area is an isotype control; the open area reflects cytokine expression. The percentage of IFN-γ and IL-4-producing iNKT cells were shown. (C) IL-4 and IFN-γ levels in the BALF were measured by ELISA. a-GC, α-GalCer. n = 5 mice for each group. *, p<0.05 using Mann-Whitney U test.</p

Anti-IL-4 Abs reduced emphysema and mucus production in mice repeatedly administered with α-GalCer.

<p>Mice were injected with neutralizing Abs to IL-4 or isotype controls 1 hour prior to each α-GalCer administration. Histopathological examination was measured 2 weeks after the last α-GalCer administration. (A) The expression of <i>Mmp-12</i> in lung tissues was analyzed by quantitative RT-PCR. (B) The expression of <i>Mmp-12</i> in lung macrophages was analyzed by quantitative RT-PCR. (C) The mean linear intercept of the alveolar septa was measured. (D) The absolute numbers of macrophages, neutrophils, eosinophils, and lymphocytes were measured. Mac, macrophage; Lym, lymphocyte; Neu, neutrophil; Eos, eosinophil. (E) The expression of <i>Muc5ac</i> in lung tissues was detected by quantitative RT-PCR. n = 9–16 mice for each group. *, p<0.05; **, p<0.01; ***, p<0.001 using Mann-Whitney U test.</p

Intranasal α-GalCer administration induced acute inflammation in the lung.

<p>Mice were intranasally administered with vehicle or α-GalCer. BALF was collected at days 0, 3 and 7. (A) BALF total cells and the absolute numbers of macrophages, lymphocytes, neutrophils, and eosinophils were measured at the indicated time points. (B) The relative expressions of cytokines and chemokines compared to positive control (set to a value of 1) of the assay kit in BALF at 3 days after α-GalCer administration of mice were determined by a dot blot immunoassay. Samples were pooled from 5–8 mice in two independent experiments. (C) Cytokines IFN-γ, IL-4, TNF-α, IL-17, and IL-13 in the BALF were assayed by ELISA. n = 5–8 mice for each group. *, p<0.05; **, p<0.01; ***, p<0.001 using Kruskal-Wallis one-way ANOVA followed Dunn's post test.</p

Increased airway inflammation in mice repeatedly administered with α-GalCer.

<p>Mice received intranasal α-GalCer administration once a week for 6 weeks and airway inflammation was measured 2 weeks after the last administration. (A) Analysis of lung function. Changes in airway resistance, dynamic compliance, and airway elastance were measured on mechanically ventilated mice, in response to increasing doses of methacholine. (B) Representative results of H-E examination of lung sections (x100 magnification). Blue arrowheads indicate cell infiltration in the lung. Black arrowheads indicate airspace enlargement in the lung. (C) The total numbers of cells present in BALF. (D) The absolute numbers of macrophages, lymphocytes, neutrophils, and eosinophils, present in BALF. Mac, macrophage; Lym, lymphocyte; Neu, neutrophil; Eos, eosinophil. (E) The absolute numbers of CD4⁺ and CD8⁺ T cells were examined. (F) CD8/CD4 ratios of T cells were measured. (G) Lung tissues were isolated and the expression of TNF-α and IL-6 was detected by quantitative RT-PCR. n = 9–14 mice for each group. (H) Macrophages from BALF were isolated and their expression of TNF-α and IL-6 was detected by quantitative RT-PCR. Individual symbols represent a sample pooled from 3–5 mice and the horizontal lines represent the mean values. a-GC, α-GalCer. *, p<0.05; **, p<0.01; ***, p<0.001 using Mann-Whitney U test.</p

IL-4 enhances <i>Mmp12</i> expression in macrophages.

<p>Bone marrow derived macrophages were stimulated with different concentrations of IL-4 or IFN-γ and the expression of <i>Mmp12</i> was measured by quantitative RT-PCR. Summarized results of 4 independent experiments are shown. Each symbol represents an individual experiment and the horizontal lines represent the means. *, p<0.05 using Kruskal-Wallis one-way ANOVA followed Dunn's post test.</p

Additional file 1 of Alteration in branching morphogenesis via YAP/TAZ in fibroblasts of fetal lungs in an LPS-induced inflammation model

Additional file 1: Figure S1. Representative immunocytochemistry staining of YAP, phosphorylated (p)-YAP, TAZ, and p-TAZ expressions in IMR-90 cells by lipopolysaccharide (LPS) at 0, 10, 30 and 50 μg/mL for 24 h. YAP and TAZ were stained in red, p-YAP and p-TAZ were stained in green, and nuclear staining were marked by DAPI in blue

The effect of AaNotch on the ultrastructure of mosquito eggs.

Mosquito eggs from control (A and A’), dsLacZ (B and B’), and dsNotch-treated (C, C’, D and D’) collected at 5 d after egg laying. Arrow: micropylar pore. Scale bar = 10 μm. (E) The percentage of eggs with complete micropylar pore formation. Number in parentheses denotes the number of eggs with complete micropylar pores divided by total number of eggs examined.</p

Demographic and cord blood data for the severe and less severe hyperbilirubinemic groups.

<p>Abbreviations: GA, gestational age; BBW, birth body weight.</p>a<p>Newborns with peak bilirubin concentration <17 mg/dL during the postnatal follow-up period.</p>b<p>Newborns with peak bilirubin concentration ≥17 mg/dL during the postnatal follow-up period.</p

The results of hepatitis B virus serology of the study population.

a<p>Anti-HBs seropositivity: defined as serum level >10 mIU/ml.</p><p>GMT: geometric mean titer.</p