Patient characteristics.

<p>*Data available for ≥80% of samples.</p><p>**Data available for ≥75% of samples.</p>a<p>Damp building-related illness.</p>b<p>Agricultural type of microbial exposure.</p>c<p>Hypersensitivity pneumonitis.</p>d<p>Sarcoidosis.</p

Immunoblot validation of protein markers in BAL.

<p>(A) Differential expression analysis based on DeCyder data for chosen protein markers. Every dot indicates the identified protein spot and its expression level in that particular study group. The difference in protein expression (average expression) between the groups is visualized with a continuous curve. Histone and semenogelin plots contains more than one curve due to several protein spot identification for histones H4, H2B (n = 3) and semenogelin I (n = 2) as shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0102624#pone.0102624.s005" target="_blank">Table S1</a>. (B) Images of immunoblot membranes and (C) immunoblot based validation results for protein markers. The number of samples in validation: CTR n = 19, AME n = 7–9, DBRI n = 17, HP n = 9, SARC n = 8. The means (black lines) are shown in scatter plots. *Indicates statistical significance, which is shown between experimental and control group, at the level of p≤0.05, **p≤0.01 and, ***p≤0.001.</p

Immunoblot validation of protein markers in plasma.

<p>The immunoblot and validation analysis for (A) galectin-3 and (B<b>)</b> histone H2B from plasma samples. The number of samples in validation: CTR n = 16, HP n = 10, SARC n = 11. The means are shown in scatter plots. *Indicates statistical significance, which is shown between experimental and control group, at the level of p≤0.05, **p≤0.01 and, ***p≤0.001.</p

Major symptoms and HRCT findings.

a<p>before BAL procedure.</p>b<p>HRCT = high-resolution computed tomography.</p>c<p>Hypersensitivity pneumonitis.</p

Reduction of <i>F</i>_ENO by tap water and carbonated water mouthwashes: magnitude and time course

<p>Fractional exhaled nitric oxide (<i>F</i>_ENO) assesses eosinophilic inflammation of the airways, but <i>F</i>_ENO values are also influenced by oral nitric oxide (NO). The aim of this pilot study was to measure <i>F</i>_ENO and compare the effect of two different mouthwashes on <i>F</i>_ENO and analyse the duration of the effect. <i>F</i>_ENO was measured in 12 randomized volunteers (healthy or asthmatic subjects) with a NIOX VERO® analyser at an expiratory flow rate of 50 mL/s. After a baseline measurement, a mouthwash was performed either with tap water or carbonated water and was measured during 20 min in 2 min intervals. The procedure was repeated with the other mouthwash. We found that both mouthwashes reduced <i>F</i>_ENO immediately at the beginning compared to the baseline (<i>p</i> < .001). The carbonated water mouthwash effect lasted 12 min (<i>p</i> ranging from <0.001 to <0.05). The tap water mouthwash reduced <i>F</i>_ENO statistically significantly only for 2 min compared with the baseline. We conclude that a single carbonated water mouthwash can significantly reduce the oropharyngeal NO contribution during a 12 min time interval.</p

The false color image of two-dimensional DIGE gel of BAL.

<p>The gel image represents the Cy3 labeled (red), Cy5 labeled (blue) and Cy2 labeled (yellow) patient samples. The latter is a pooled sample, which served as an internal standard. Spot abbreviations refer to the identified proteins listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0102624#pone.0102624.s005" target="_blank">Table S1</a>. Molecular weights are shown on the right edge of the gel and the pI range on the top part of the SDS-PAGE.</p

Exercise is associated with younger methylome and transcriptome profiles in human skeletal muscle

Exercise training prevents age-related decline in muscle function. Targeting epigenetic aging is a promising actionable mechanism and late-life exercise mitigates epigenetic aging in rodent muscle. Whether exercise training can decelerate, or reverse epigenetic aging in humans is unknown. Here, we performed a powerful meta-analysis of the methylome and transcriptome of an unprecedented number of human skeletal muscle samples (n = 3176). We show that: (1) individuals with higher baseline aerobic fitness have younger epigenetic and transcriptomic profiles, (2) exercise training leads to significant shifts of epigenetic and transcriptomic patterns toward a younger profile, and (3) muscle disuse “ages” the transcriptome. Higher fitness levels were associated with attenuated differential methylation and transcription during aging. Furthermore, both epigenetic and transcriptomic profiles shifted toward a younger state after exercise training interventions, while the transcriptome shifted toward an older state after forced muscle disuse. We demonstrate that exercise training targets many of the age-related transcripts and DNA methylation loci to maintain younger methylome and transcriptome profiles, specifically in genes related to muscle structure, metabolism, and mitochondrial function. Our comprehensive analysis will inform future studies aiming to identify the best combination of therapeutics and exercise regimes to optimize longevity