Human bronchial epithelial cells were exposed to TDI or vehicle. Results TDI-exposed mice had higher numbers of airway goblet cells, BAL eosinophils, CD4+ T cells and ILC, with a predominant type 2 response and tended to have airway hyperresponsiveness. In TDI-exposed microRNA-155 knockout mice, inflammation and airway hyperresponsiveness was attenuated. TDI exposure induced IL-33 expression in human bronchial epithelial cells and in murine lungs, which was microRNA-155 dependent in mice. GATA3+CD3- cells, presumably ILC2, were present in bronchial biopsies. Conclusion TDI exposure is associated with increased numbers of ILC. The proinflammatory microRNA-155 is crucial in a murine model of TDI asthma, suggesting its involvement in the pathogenesis of occupational asthma due to LMW agents.Introduction Diagnosing asthma in children remains a challenge because respiratory symptoms are not specific and vary over time. Aim In a real-life observational study, we assessed the diagnostic accuracy of respiratory symptoms, objective tests, and two paediatric diagnostic algorithms proposed by GINA and NICE to diagnose asthma in school-aged children. Methods We studied children aged 5-17 years referred consecutively for evaluation of suspected asthma to pulmonary outpatient clinics. Symptoms were assessed by parental questionnaire. The investigations included specific IgE measurement or skin prick tests, measurement of fractional exhaled nitric oxide, spirometry, body plethysmography, and bronchodilator reversibility. Asthma was diagnosed by paediatric pulmonologists based on all available data. We assessed diagnostic accuracy of symptoms, tests, and diagnostic algorithms by calculating sensitivity, specificity, positive and negative predictive values, and area under the curve (AUC). Results Among 514 participants, 357(70%) were diagnosed with asthma. The combined sensitivity and specificity (sensitivity/specificity) was highest for any wheeze (0.75/0.65), dyspnoea (0.56/0.76), and wheeze triggered by colds (0.58/0.78) or by exercise (0.55/0.74). Of the diagnostic tests, the AUC was highest for specific total resistance (sRtot) (0.73) and lowest for the residual volume (RV) total lung capacity (TLC) ratio (0.56). The NICE algorithm had a sensitivity of 0.69 and specificity of 0.67, whereas the GINA algorithm had a sensitivity of 0.42 and specificity of 0.90.  https://www.selleckchem.com/products/gsk3787.html  Conclusion This study confirms the limited usefulness of single tests as well as existing algorithms for the diagnosis of asthma. It highlights the need for new and more appropriate evidence-based guidance.Introduction Neutrophilic inflammation is a major driver of bronchiectasis pathophysiology, and neutrophil elastase activity is the most promising biomarker evaluated in sputum to date. How active neutrophil elastase correlates with lung microbiome in bronchiectasis is still unexplored. We aimed at understanding if active neutrophil elastase is associated with low microbial diversity and distinct microbiome characteristics. Methods An observational, cross-sectional study was conducted at the Bronchiectasis Program of the Policlinico Hospital in Milan, Italy, where adults with bronchiectasis were enrolled between March 2017 and March 2019. Active neutrophil elastase was measured on sputum collected during stable state, microbiota analysed through 16S rRNA gene sequencing, molecular assessment of respiratory pathogens through real time PCR and clinical data collected. Measurements and main results Among 185 patients enrolled, decreasing alpha diversity, evaluated through the Shannon entropy (rho -0.37; p-value less then 0.00001), Pielou' evenness (rho -0.36, p less then 0.00001) and richness (rho -0.33; p less then 0.00001), was significantly correlated with increasing elastase. A significant difference in median levels of Shannon was detected between patients with neutrophil elastase ≥20 µg·mL-1 [3.82 (2.20-4.96)] versus neutrophil elastase less then 20 µg·mL-1 [4.88 (3.68-5.80)], p less then 0.0001. A distinct microbiome was found in these two groups, mainly characterised by enrichment with Pseudomonas in the high and with Streptococcus in the low elastase group. Further confirmation of the association of P. aeruginosa with elevated active neutrophil elastase was found based on standard culture and targeted real-time PCR. Conclusions High levels of active neutrophil elastase are associated to low microbiome diversity and specifically to P. aeruginosa infection.Alcohol dehydrogenases (ADHs) and aldehyde dehydrogenases (ALDHs) are vital enzymes involved in the metabolism of a variety of alcohols. Differences in the expression and enzymatic activity of human ADHs and ALDHs correlate with individual variability in metabolizing alcohols and drugs and in the susceptibility to alcoholic liver disease. MicroRNAs (miRNAs) function as epigenetic modulators to regulate the expression of drug-metabolizing enzymes. To characterize miRNAs that target ADHs and ALDHs in human liver cells, we carried out a systematic bioinformatics analysis to analyze free energies of the interaction between miRNAs and their cognate sequences in ADH and ALDH transcripts and then calculated expression correlations between miRNAs and their targeting ADH and ALDH genes using a public data base. Candidate miRNAs were selected to evaluate bioinformatic predictions using a series of biochemical assays. Our results showed that 11 miRNAs have the potential to modulate the expression of two ADH and seven ALDH genes in the human liver. We found that hsa-miR-1301-3p suppressed the expression of ADH6, ALDH5A1, and ALDH8A1 in liver cells and blocked their induction by ethanol. In summary, our results revealed that hsa-miR-1301-3p plays an important role in ethanol metabolism by regulating ADH and ALDH gene expression. SIGNIFICANCE STATEMENT Systematic bioinformatics analysis showed that 11 microRNAs might play regulatory roles in the expression of two alcohol dehydrogenase (ADH) and seven aldehyde dehydrogenase (ALDH) genes in the human liver. Experimental evidences proved that hsa-miR-1301-3p suppressed the expression of ADH6, ALDH5A1, and ALDH8A1 in liver cells and decreased their inducibility by ethanol.