L-NAME-induced senescence in lung tissue.

<p><b>(A)</b> ATLR measurements from 8 week-treated lungs. <b>(B-D)</b> qRT-PCR data from 1 week-treated lungs evaluating the senescence markers <b>(B)</b> p16^Ink4a, <b>(C)</b> p53, and <b>(D)</b> p21. <b>(A)</b> *P = 0.007. n = 12. <b>(B-D)</b> *P = 0.02. n = 7–11. Data are mean ± SD.</p

Effects of L-NAME on lung functional dynamics.

<p>Measurements for <b>(A)</b> compliance, <b>(B)</b> static compliance, <b>(C)</b> elastance, and <b>(D)</b> static elastance demonstrate that L-NAME-treated lungs have the functional characteristics of emphysema. Both genetic and pharmacologic inhibition of PAI-1 protected mice from lung dysfunction. <b>(A)</b> and <b>(C)</b> *P = 0.002, #P = 0.02, $P = 6.3x10^-6. <b>(B)</b> and <b>(D)</b> *P = 0.004, #P = 0.01. Data are mean ± SD. n = 6–7.</p

Alcohol accelerates mortality from hyperoxia-induced acute lung injury in mice.

<p>Mice were treated with ethanol (20% v/v, 4 g/kg i.p.) or control vehicle (sterile water) daily starting 3 days prior to and during the first 2 days of exposure to hyperoxia (>95% O₂) for a total of 5 days. The time when 50% of the animals had died (LD₅₀) was calculated. (N = 8/group for each treatment group).</p

Alcohol decreases the rate of alveolar fluid clearance and downregulates Na,K-ATPase <i>in vivo</i> and <i>in vitro</i>.

<p>Mice were treated with ethanol (20%, 4 g/kg i.p.) or control vehicle (sterile w) daily for 5 days and (<b>A</b>) alveolar fluid clearance was measured and (<b>B</b>) basolateral membrane (BLM) abundance of α₁-subunit of the Na,K-ATPase in peripheral lung tissue was evaluated via immunoblotting 4 hours after the last dose. N = 7 mice/group for alveolar fluid clearance measurements and N = 3 for evaluation of Na,K-ATPase, *p<0.05 between alcohol and control vehicle. (<b>C, D</b>) Plasma membrane abundance of the α₁-subunit of the Na,K-ATPase in primary rat AEC was evaluated via immunoblotting (<b>C</b>) 30 minutes after treatment with different doses of ethanol and (<b>D</b>) for up to 30 minutes after treatment with high dose ethanol (100mM). N≥3 for all measures, *p<0.05 for comparison with untreated cells.</p

L-NAME treatment causes emphysema.

<p>Lung tissue sections from <b>(A)</b> WT, <b>(B)</b> WT + L-NAME, <b>(C)</b> WT + L-NAME + TM5441, and <b>(D)</b> PAI-1-/- + L-NAME demonstrate that L-NAME causes significant alveolar destruction and that PAI-1 inhibition is partially protective against this. <b>(E)</b> Mean linear intercept quantifications. *P = 0.0002, #P = 0.04, $P = 0.009. Data are mean ± SD. n = 11–13.</p

Alcohol does not increase ROS generation in alveolar epithelial cells.

<p>(<b>A</b>) MLE-12 cells that stably express a mitochondrially localized oxidant sensitive GFP probe were treated with different doses of ethanol (0-100 mM), media (negative control) or H₂O₂ (positive control) and ROS generation was measured 4 hours later. (<b>B</b>) The same cells were treated with ethanol (100 mM) or media (control), and ROS generation was measured at different time points. N≥3 for all measures, *p<0.05 for comparison with untreated cells.</p

Alcohol-induced reduction in Na,K-ATPase is mediated via phosphorylation of AMP-activated protein kinase.

<p>Immunoblotting for phosphorylated AMPK (p-AMPK) and total AMPK was performed in cell lysates from (<b>A</b>) AEC 15 minutes after treatment with different concentrations of ethanol, H₂O₂ (positive control) or media (negative control), (<b>B</b>) at different time points after the administration of ethanol (100 mM) or media or (<b>C</b>) after treatment with a ADORA1 agonist (CPA, 10⁻⁵M) or control (DMSO). (<b>D</b>) Membrane abundance of the α₁-subunit of Na,K-ATPase was evaluated via immunoblotting in rat AEC transfected with an adenovirus that expresses no transgene (Adnull) or that expresses a dominant-negative, kinase-dead variant of the AMPK α₁ subunit (AdDN AMPK) to generate a non-functional AMPK. N≥3 for all measures, *p<0.05 for comparison with untreated cells.</p

Rosuvastatin does not alter the clinical course of influenza A infection in mice.

<p>We treated mice with rosuvastatin or control therapy starting 3 days before they were infected with either Udorn or WSN strains of influenza A virus and measured (<b>A</b>) WSN-associated mortality and (<b>B</b>) Udorn- and (<b>C</b>) WSN-associated changes in daily weight.</p

Alcohol increases adenosine levels in the lung and induces the release of adenosine from primary alveolar epithelial cells.

<p>(<b>A</b>) We treated mice with either ethanol (20% v/v, 4 g/kg, i.p.) or control vehicle (sterile water) and adenosine levels in BAL fluid were measured at 4 hours using HPLC. (<b>B</b>) We exposed mice to either hyperoxia (>95% O₂) or normoxia (21%) for 72 hours and then measured adenosine levels in BAL fluid. Primary rat alveolar epithelial cells were treated with ethanol and (<b>C</b>) adenosine levels in the culture media (<b>D</b>) ADP and ATP levels in cell lysates were measured after 4 hours. N ≥ 3 for all measures, *p<0.05 for comparison with untreated cells.</p

Rosuvastatin does not affect the influenza A-induced changes in pro-inflammatory cytokines in the lungs.

<p>We treated mice with rosuvastatin or control starting 3 days before they were infected with either Udorn or WSN strains of influenza A virus. Four days after influenza A infection, we collected bronchoalveolar lavage fluid (BALF) and measured (<b>A, B</b>) TNF-α and (<b>C, D</b>) IL-6 levels. *P<0.05 Udorn vs. PBS, WSN vs. PBS. NS; not significant (Rosuvastatin vs. Control treatment).</p