Culture & Childhood Obesity: Investigating maternal experiences

<p>(A) Perivascular and (B) bronchial inflammation was recorded in mice exposed to cigarette smoke and RSV for 6 months and their corresponding controls. (C) Matrix accumulation was assessed with trichrome staining in each mouse group and quantified by the Ashcroft fibrosis score. Representative images of mice lungs from each group are presented here (scale bar = 20 µM; left panels). Fibrosis and inflammation scores were calculated for each treatment group (right panels where n = 12 animals/group). Each graph is represented as mean ± S.E.M. where each measurement was performed on 12 animals/group. p values shown, comparing both treatments connected by a line.</p

RSV infections alter cigarette smoke induced airway cytokine gene expressions.

<p>Values are represented as mean ± S.E.M., where each measurement was performed 3 times on 12 animals/group. Bold numbers denoted by *represents a p value less than 0.05 compared to mock and room air treated mice.</p>#<p>denotes a p value less than 0.05 compared to either smoke or RSV treated mice.</p

RSV infections alter cigarette smoke induced airway protease gene expressions.

<p>Values are represented as mean ± S.E.M., where each measurement was performed 3 times on 12 animals/group. Bold numbers denoted by *represents a p value less than 0.05 compared to mock and room air treated mice.</p>#<p>denotes a p value less than 0.05 compared to either smoke or RSV treated mice.</p

RSV infections enhance cigarette smoke induced airway cell death.

<p>(A) TUNEL analysis was performed on lung tissue from mice exposed to cigarette smoke and RSV for 6 months and their corresponding controls. Graph represented as mean ± S.E.M., where each measurement was performed on 12 animals/group. p values shown, comparing both treatments connected by a line. (B) Representative images of TUNEL staining of mice lungs from each group are presented here (scale bar = 50 µM).</p

Smoke enhances RSV inhibition of PTP1B and PP2A activities.

<p>Lung tissue from C57BL/6J mice was examined for gene expression and phosphatase activity for (A) PP2A and (B) PTP1B. Graphs are represented as mean ± S.E.M., where each measurement was performed on 12 animals/group. p values shown, comparing both treatments connected by a line.</p

Chronic cigarette smoke exposure enhances RSV pathology in mouse airways.

<p>(A) Protocol for administering A2 RSV strain and cigarette smoke to C57BL/6J mice. C57BL/6J mice (n = 12 animals/group) were infected with monthly RSV infections in combination with 6 months of smoke exposure. (B) Changes in body weight of each treatment group of C57BL/6J mice, represented as a percentage of initial weight of mock and room air treated mice at the beginning of this study. p values shown, comparing both groups by 2-way ANOVA. (C) RSV N copy number was determined 10 days post final infection in the lungs of C57BL/6J mice infected with 1×10⁶ pfu of RSV A2 strain and exposed to either room air or cigarette smoke for 6 months. Absolute RSV N concentration was represented as natural log pg. (D) BALF immune cellularity was measured and changes were observed following repeat monthly RSV infections in combination with 6 months of smoke exposure on total immune cell number, macrophages, neutrophils and lymphocytes. Graphs are represented as mean ± S.E.M., where each measurement was performed 3 times on 12 animals/group. p values shown, comparing both treatments connected by a line.</p

RSV infections enhance cigarette smoke induced cathepsin S activity.

<p>BALF protease activity was of mice exposed to cigarette smoke and RSV for 6 months and their corresponding controls. (A) Total BALF collagenase and cathepsin S relative activity was determined. (B) Gelatinase activity was determined in BALF and densitometry was performed for MMP-9 and MMP-2. Graphs are represented as mean ± S.E.M., where each measurement was performed 3 times on 12 animals/group. p values shown, comparing both treatments connected by a line.</p

LL-37 inhibits LPS-induced IL-8 production and IκB degradation in THP-1 monocytic cells.

<p>(<b>A</b>) THP-1 monocytes were pre-treated for 1 h with 10 µg/ml LL-37 and incubated in the absence or presence of 1 µg/ml <i>P. aeruginosa</i> LPS. After 24 h stimulation with LPS, IL-8 levels were quantified in cell-free culture supernatants by ELISA. *** <i>p</i><0.001; Con, control. (<b>B</b>) Cytoplasmic extracts were prepared following 30, 60 and 120 min and levels of IκBα, IκBβ, phosphorylation of IκBα (Ser32/36) and IKKα/β (Ser180/181) were determined by Western blotting. GAPDH was used to control for protein loading.</p

Increased LL-37 in DNase and heparinase II treated CF sputum inhibits IL-8 production from THP-1 monocytes.

<p>CF sputum samples (n = 12) were treated with Pulmozyme® DNase and/or heparinase II for 1 h at 37°C in the absence or presence of an LL-37 antibody or heat-inactivated (HI) LL-37 antibody as a negative control. Samples were incubated in LPS-immobilised 96-well microtiter plates (100 ng LPS/well) for 1 h at RT. Unbound peptides were removed by washing and THP-1 monocytes (1×10⁶/ml) were added to each well and incubated for 6 h at 37°C. The ability of released LL-37 to inhibit LPS-induced IL-8 production by THP-1 monocytes was determined by ELISA. Results are expressed as percentage of untreated control (100% = untreated CF sputum). *** <i>p</i><0.001.</p

LL-37 levels and its ability to bind LPS in CF sputum are increased following treatment with DNase and heparinase II.

<p>(<b>A</b>) CF sputum samples (n = 12) were treated with Pulmozyme® DNase and/or heparinase II for 1 h at 37°C. The LL-37 content of these samples and untreated sputum samples were quantified by ELISA. (<b>B</b>) The ability of available LL-37 present in untreated and treated CF sputum samples to bind LPS was determined by ELISA. Results are expressed as percentage of untreated control (100% = untreated CF sputum). ** <i>p</i><0.01; *** <i>p</i><0.001.</p