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Donato Lacedonia,* Grazia Pia Palladino,* Maria Pia Foschino-Barbaro, Giulia Scioscia, Giovanna Elisiana Carpagnano Department of Medical and Surgical Sciences, Institute of Respiratory Diseases, University of Foggia, Foggia, Italy *These authors contributed equally to this work Background and objectives: A new phenotype with overlapping characteristics between asthma and chronic obstructive pulmonary disease (COPD) called asthma–COPD overlap syndrome (ACOS) is emerging among inflammation diseases. To date, there is no agreement on specific criteria to define this syndrome, and the current guidelines are insufficient to classify the analogy and differences between overlap and COPD or asthma phenotypes. It would be necessary to identify new biomarkers able to identify these diseases clearly. Thus, the aim of this study was to identify a serum and supernatant of sputum microRNA (miRNA) expression profile of miRNA-145 and miRNA-338 in patients with asthma (n=13), COPD (n=31), and ACOS (n=8) and controls (n=7). Methods: The expression was evaluated using quantitative real-time polymerase chain reaction (qRT-PCR). For statistical analysis, the ANOVA test, Kruskal–Wallis test, Mann–Whitney U-test, and Spearman’s rank correlation were used. Results: The main finding of this work is that the expression of miRNA-338 is higher in the supernatant of different obstructive diseases than in peripheral blood, while miRNA-145 is higher only in the supernatant of asthma patients. The expression of both selected miRNAs is higher in the supernatant of asthma and COPD patients than in controls. Conclusion: Differences in sputum miRNA expression profile were observed between patients with ACOS and asthma or COPD, which underline the potential role of miRNA as a biomarker that is able to discriminate patients with ACOS, asthma, and COPD. Keywords: asthma–COPD overlap syndrome, ACOS, asthma, chronic obstructive pulmonary disease, miRNA, sputum

Donato Lacedonia,1,* Giovanna Elisiana Carpagnano,1,* Teresa Trotta,2 Grazia Pia Palladino,1 Maria Antonietta Panaro,3 Liugi Davide Zoppo,1 Maria Pia Foschino Barbaro,1 Chiara Porro21Department of Medical and Surgical Sciences, Institute of Respiratory Diseases, 2Department of Clinical and Experimental Medicine, University of Foggia, Foggia, 3Department of Biosciences, Biotechnologies and Biopharmaceutics, University of Bari, Bari, Italy*These authors contributed equally to this workBackground: Microparticles (MPs) are small membrane vesicles of 0.1–1 µm which are released by cells following chemical, physical, and apoptotic stimuli. MPs represent more than a miniature version of the cell. Their composition and function depend not only on cellular origin, but also on stimuli. Chronic obstructive pulmonary disease (COPD) is a lung disease characterized by nearly irreversible lung destruction which results in airway limitation.Purpose: We investigated the presence and source of MPs in sputum of COPD patients to evaluate if changes in MP number and origin may reflect the pathophysiological conditions of disease and may serve as potential biomarkers for diagnostic and prognostic use.Methods: Induced sputum samples were collected from 18 male subjects and liquefied with Sputasol. MPs obtained were immunolabeled for leukocyte (CD11a), granulocyte (CD66b), monocyte-macrophage (CD11b), platelets and megakaryocytic cells (CD41), endothelial cells (CD31), and red blood cells (CD235ab) and analyzed by cytofluorimetry.Results: There was a negative correlation between CD31-MPs and forced expiratory volume in 1 second (R=−53, P

Background: Wiedemann-Steiner syndrome (WSS), a rare autosomal-dominant disorder caused by haploinsufficiency of the KMT2A gene product, is part of a group of disorders called chromatinopathies. Chromatinopathies are neurodevelopmental disorders caused by mutations affecting the proteins responsible for chromatin remodeling and transcriptional regulation. The resulting gene expression dysregulation mediates the onset of a series of clinical features such as developmental delay, intellectual disability, facial dysmorphism, and behavioral disorders. Aim of the Study: The aim of this study was to investigate a 10-year-old girl who presented with clinical features suggestive of WSS. Methods: Clinical and genetic investigations were performed. Whole exome sequencing (WES) was used for genetic testing, performed using Illumina technology. The bidirectional capillary Sanger resequencing technique was used in accordance with standard methodology to validate a mutation discovered by WES in all family members who were available. Utilizing computational protein modeling for structural and functional studies as well as in silico pathogenicity prediction models, the effect of the mutation was examined. Results: WES identified a de novo heterozygous missense variant in the KMT2A gene KMT2A(NM_001197104.2): c.3451C>G, p.(Arg1151Gly), absent in the gnomAD database. The variant was classified as Likely Pathogenetic (LP) according to the ACMG criteria and was predicted to affect the CXXC-type zinc finger domain functionality of the protein. Modeling of the resulting protein structure suggested that this variant changes the protein flexibility due to a variation in the Gibbs free energy and in the vibrational entropy energy difference between the wild-type and mutated domain, resulting in an alteration of the DNA binding affinity. Conclusions: A novel and de novo mutation discovered by the NGS approach, enhancing the mutation spectrum in the KMT2A gene, was characterized and associated with WSS. This novel KMT2A gene variant is suggested to modify the CXXC-type zinc finger domain functionality by affecting protein flexibility and DNA binding.

Idiopathic pulmonary fibrosis (IPF) is a chronic debilitating fibrotic lung disease leading to respiratory failure and ultimately to death. Noninvasive biomarkers, for the early diagnosis, differential diagnosis, prognosis, and prediction of therapeutic response, are needed. Previous studies support a role for periostin in lung fibrosis. The aim of our study was to analyze periostin levels in the airways of patients with IPF and to investigate its role as a useful predictive biomarker of the disease. We enrolled 30 IPF patients and 5 control subjects. All subjects underwent all standard radiological, functional, and biological examinations for IPF diagnosis and staging and exhaled breath condensate (EBC) collection. Periostin was assessed by an enzyme-linked immunosorbent assay kit on EBC. Periostin was dosable in the EBC of all subjects enrolled. We found higher exhaled periostin levels in IPF patients than healthy controls (65.5 ± 23.5 pg/mL vs. 33 ± 21.4 pg/mL, p  < 0.05). Moreover, in receiver operating characteristic analysis, the clinical reference value of periostin was 37.88 pg/mL to discriminate patients with IPF from healthy subjects, with the area under the curve of 0.8815. There was no significant correlation between periostin levels and gender or pulmonary function tests. These preliminary results support our working hypothesis that periostin is dosable in the airways of patients with IPF. As the circulating periostin, also airways periostin may be a potential biomarker to support IPF diagnosis and to monitor disease progression during follow-up.

Background: Idiopathic Pulmonary Fibrosis (IPF) is a degenerative interstitial lung disease with both a poor prognosis and quality of life once the diagnosis is made. In the last decade many features of the disease have been investigated to better understand the pathological steps that lead to the onset of the disease and, moreover, different types of biomarkers have been tested to find valid diagnostic, prognostic and therapy response predictive ones. In the complexity of IPF, microRNA (miRNAs) biomarker investigation seems to be promising., Methods: We analysed the expression of five exosomal miRNAs supposed to have a role in the pathogenesis of the disease from serum of a group of IPF patients (n = 61) and we compared it with the expression of the same miRNAs in a group of healthy controls (n = 15)., Results: In the current study what emerged is let-7d down-regulation and, unexpectedly, miR-16 significant down-regulation. Moreover, through a cross-sectional analysis, a clustering of the expression of miR-16, miR-21 and miR-26a was found., Conclusions: These findings could help the individuation of previously unknown key players in the pathophysiology of IPF and, most interestingly, more specific targets for the development of effective medications.

Introduction: Several studies have previously demonstrated that long-term exposure to outdoor pollution present airway inflammation in term of an increase of sputum neutrophils., Aim and Methods: The aim of our study was to evaluate the level of airway inflammation by induced sputum in a group of 15 non-professionally exposed population of well-characterized COPD patients, residing in urban areas with high rate of outdoor pollution, compared to a control group of 13 individuals with COPD, living in rural areas with a low pollution rate. All participants underwent spirometry and sputum induction., Results: A statistically significant increase in the percentage of neutrophil cell count was found among the residents in urban areas compared to those living in rural regions (89.1 vs 79.0, p<0.05) CONCLUSIONS: In conclusion, we showed that non-professionally exposed patients with COPD residing in highly-polluted urban areas had greater airway inflammation in terms of sputum neutrophils compared to a population with very similar characteristics, living in rural areas with lower outdoor pollution. The results of this pilot study may be relevant for the long term effect of environmental outdoor pollution in vulnerable patients like those with COPD., (Copyright © 2019 SEPAR. Publicado por Elsevier España, S.L.U. All rights reserved.)

Background: Obstructive Sleep Apnea (OSAS) is a disease associated with the increase of cardiovascular risk and it is characterized by repeated episodes of Intermittent Hypoxia (IH) which inducing oxidative stress and systemic inflammation. Mitochondria are cell organelles involved in the respiratory that have their own DNA (MtDNA). The aim of this study was to investigate if the increase of oxidative stress in OSAS patients can induce also MtDNA alterations., Methods: 46 OSAS patients (age 59.27 ± 11.38; BMI 30.84 ± 3.64; AHI 36.63 ± 24.18) were compared with 36 control subjects (age 54.42 ± 6.63; BMI 29.06 ± 4.7; AHI 3.8 ± 1.10). In blood cells Content of MtDNA and nuclear DNA (nDNA) was measured in OSAS patients by Real Time PCR. The ratio between MtDNA/nDNA was then calculated. Presence of oxidative stress was evaluated by levels of Reactive Oxygen Metabolites (ROMs), measured by diacron reactive oxygen metabolite test (d-ROM test)., Results: MtDNA/nDNA was higher in patients with OSAS than in the control group (150.94 ± 49.14 vs 128.96 ± 45.8; p = 0.04), the levels of ROMs were also higher in OSAS subjects (329.71 ± 70.17 vs 226 ± 36.76; p = 0.04) and they were positively correlated with MtDNA/nDNA (R = 0.5, p < 0.01)., Conclusions: In OSAS patients there is a Mitochondrial DNA damage induced by the increase of oxidative stress. Intermittent hypoxia seems to be the main mechanism which leads to this process.

Background and Aims: Lung cancer has recently been associated with human papilloma virus (HPV) infection. The most important event associated with HPV infection in cancer foresees HPV DNA integration into the host genome. Sites of integration such as the fragile site FRA3B adjacent to the FHIT frequently undergo microsatellite alterations (MAs). In this study we aim to verify the role of MAs at 3p in non-small cell lung cancer (NSCLC) with HPV positivity and eventual correlation with sex, histotype, TNM stage and cigarette smoking., Methods: We enrolled 26 NSCLC patients previously investigated for the presence of HPV in their airways (11 HPV+ and 15 HPV-). All subjects had allelotyping analysis of DNA from exhaled breath condensate (EBC), blood and bronchial brushing of microsatellite D3S1300 located in the chromosomal region 3p., Results: For the first time we described the presence of MAs at 3p in EBC of NSCLC patients with HPV positivity. MAs in EBC corresponded to those in paired brushing. The number of patients with 3p MAs was higher in the group of NSCLC with HPV positivity than with HPV negativity. No relationship between the presence and type of MAs in EBC-brushing/DNA and sex, histotype or tumor stage was found., Conclusion: Our results suggested that MAs at 3p are present in caucasic NSLC HPV+ patients and might be involved in lung carcinogenesis. In consideration of the possible clinical usefulness of the analysis of MAS at 3p in the EBC of HPV+ patients in the non-invasive screening for lung cancer, these results merit further studies., (Copyright © 2014 IMSS. Published by Elsevier Inc. All rights reserved.)

Background: Airways of lung cancer patients are often colonized by fungi. Some of these colonizing fungi, under particular conditions, produce cancerogenic mycotoxins. Given the recent interest in the infective origin of lung cancer, with this preliminary study we aim to give our small contribution to this field of research by analysing the fungal microbiome of the exhaled breath condensate of lung cancer patients from Puglia, a region of Italy., Methods: We enrolled 43 lung cancer patients and 21 healthy subjects that underwent exhaled breath condensate and bronchial brushing collection. The fungal incidence and nature of sample collected were analysed by using a selected media for Aspergillus species., Results: For the first time we were able to analyse the fungal microbioma of the exhaled breath condensate. 27.9% of lung cancer patients showed a presence of Aspergillus niger, or A. ochraceus or Penicillium ssp. while none of the healthy subjects did so., Conclusion: The results confirmed the high percentage of fungal colonization of the airways of lung cancer patients from Puglia, suggesting the need to conduct further analyses in this field in order to evaluate the exact pathogenetic role of these fungi in lung cancer as well as to propose efficient, empirical therapy.

Background: The aim of this study was to define the involvement of some biomarkers in patients with chronic obstructive pulmonary disease (COPD) and pulmonary hypertension (PH), with particular attention to sub-groups with a PH that is "out of proportion" (OP)., Materials and Methods: Patients with COPD without PH, with PH and marked airways obstruction, and with PH and mild airways obstruction were compared. Assays for human interleukin-6 (IL-6), leukotriene B4 (LTB4), vasoactive intestinal peptide (VIP), and endothelin-1 (ET-1) were performed on the blood samples taken during right heart catheterization (RHC) in a pulmonary artery., Results: In all, 83 patients were enrolled and divided into three groups: 37 simple COPD (mean pulmonary artery pressure [mPAP] <25 mmHg) and 46 COPD with PH (mPAP ≥25 mmHg). Among the latter, those who had a mPAP ≥35 mmHg and forced expiratory volume in 1 sec [FEV1] ≥50% were classified as OP (7 patients). Patients with PH were older and had a body mass index (BMI) higher than the other groups; moreover, they had lower FEV1 and carbon monoxide diffusion (DLCO) values. A lower level of partial pressure of oxygen in arterial blood (PaO2) was observed in the group of OP patients. The levels of ET-1, IL-6, and LTB4 were similar in each group; VIP was higher in the OP patients than in simple COPD and was related to PAP., Conclusions: In the patients with COPD and PH and in particular in the group of OP PH, VIP is significantly increased, probably to correct the imbalance between vasoconstrictor and vasodilatator mediators.

Background and Objectives: Airway remodeling is a main feature of asthma. Different biological phenotypes of severe asthma have been recently recognized by the ENFUMOSA study group and among these one is characterized by neutrophilic airway inflammation. Concentrations of MMP-9 in airways have been suggested as a marker to monitor airway remodeling in asthma., Objective: The aim of the present study was to explore airway remodeling in different biological phenotypes of asthma by measuring MMP-9 in EBC and correlating these with other variables., Methods: Sixty consecutive subjects with asthma and 20 healthy controls were enrolled in the study. Exhaled MMP-9, pH and NO levels and inflammatory cells in sputum were measured in all subjects enrolled., Results: We observed an increase of exhaled MMP-9 in asthmatic subjects compared to controls. Higher exhaled MMP-9 concentrations were described in severe asthmatics compared to mild to moderate especially in those with neutrophilic airway inflammation. We further found a correlation between exhaled MMP-9 and percentage of neutrophils in sputum, FEV1, exhaled NO and pH., Conclusion: Our results seem to substantiate the feasibility of measuring exhaled MMP-9 in the breath of asthmatic patients. MMP-9 may be considered a proxy of the amount of the ongoing airway remodeling in asthma. MMP-9 has been shown to be differentially released in different phenotypes of asthma. The measure of exhaled MMP-9 could help to monitor the ongoing airway remodeling, recognize severe stages of asthma, and possibly help determine the appropriate choice of therapy., (© 2013.)

Aim: The aim of the study was to investigate the effect of selective ETRA Sitaxsentan on viability and differentiation into myofibroblasts of lung fibroblasts derived from SSc-ILD patients and the ability of this drug to modify the lung fibroblast synthesis of VEGF, type I collagen and fibronectin., Methods: Primary human lung fibroblast cultures were obtained from BAL of SSc-ILD patients. Cell cultures were exposed for 48 h to crescent concentrations of Sitaxsentan (10 -6M to 10 -4M). In these experimental conditions we evaluated cell viability through crystal violet staining, the production and mRNA expression of VEGF, fibronectin and type I collagen respectively through ELISA and real-Time PCR. Further, we detected alpha-Smooth Muscle Actin (α-SMA) through immunocytochemical assay., Results: The lowest concentration of sitaxsentan (10-6M) did not affect fibroblasts viability; conversely at higher concentrations, sitaxsentan induced a significant inhibition of cell viability. Synthesis and mRNA expression of VEGF, type 1 collagen and fibronectin were significantly reduced in treated lung fibroblasts compared to the untreated ones, in a dose-dependent manner. At higher concentrations, Sitaxsentan reduced the expression of α-SMA., Conclusion: The results of this study show that sitaxentan is able in vitro to reduce both cell viability than production of VEGF and extra-cellular matrix components in SSc lung fibroblasts, confirming the anti-fibrotic potential of ETRA in SSc. The decreased expression of α-SMA in treated cells indicate that sitaxsentan may inhibit the fibroblast differentiation toward a myo-fibroblast phenotype and further support the hypothesis that the selective ETRAs may be beneficial in patients with SSc-ILD as anti fibrotic agents.

Background: Since little is known of airways inflammation in the elderly, we have carried out a study to explore the presence of some inflammatory markers in the airways of healthy subjects of different ages using a non-invasive method which is particularly suitable for aged people., Objective: The aim of this work was to investigate whether parameters, including (1) pH, IL-8 and TNF-α in exhaled breath condensate (EBC), (2) exhaled nitric oxide levels (NO), and (3) inflammatory cell profile in induced sputum, are age-related., Materials and Methods: Thirty healthy adults (10 subjects below the age of 30 [A], 10 subjects between 30 and 60 years [B], and 10 subjects over 60 years of age [C]), were enrolled in the study. IL-8 and TNF-α levels were measured in breath condensate. Exhaled pH was measured after deaeration/decarbonation by means of a pH-meter. A rapid-response chemiluminescence NO analyzer was used to quantify NO. Induced sputum was collected, homogenized with dithiothreitol, and cytospins for differential cell were produced., Results: The levels of IL-8 and TNF-α in EBC, the levels of exhaled NO, and the percentage of neutrophils in induced sputum were significantly elevated in C and B compared with A; the EBC pH level was significantly reduced in C and B compared with A. The EBC levels of IL-8, TNF-α, pH, the level of exhaled NO, and the percentage of neutrophils correlated significantly with age., Conclusion: This study has shown the presence of age-related airways inflammation in healthy subjects.

Background: Matrix metalloproteinase-9 (MMP-9) has been recognized in several types of tumor development and progression, including lung cancer, for its role in the degradation and remodeling of lung tissue. Furthermore, increased MMP-9 has been commonly described in the serum and airways of non-small cell lung cancer (NSCLC) patients., Objective: The aim of this study was to investigate, for the first time, MMP-9 in the exhaled breath condensate (EBC) of NSCLC patients., Participants: We enrolled 40 NSCLC patients and 40 controls affected by transudative pleural effusion., Measurements: MMP-9 concentrations were measured in the EBC, whole blood (WB), and pleural effusion (PE) of all the subjects under study using enzyme immunoassay (EIA) kits., Results: MMP-9 levels were found to be significantly higher in EBC, WB, and PE of NSCLC patients compared with controls. A positive correlation was observed between MMP-9 in EBC, cigarettes smoked, and stage of cancer., Conclusion: Exhaled MMP-9 was elevated in NSCLC patients, especially during tumor progression, and could represent a suitable noninvasive marker in the diagnosis and monitoring of lung cancer.

Background: Today an increasing interest is being generated by the study of lung cancer markers in the exhaled breath condensate (EBC), precisely because this sample seems to lend itself to lung cancer early screening and follow-up. Indeed, ferritin and superoxide dismutase (SOD) have recently been recognized to play a role in lung cancerogenesis and patients' survival. The aim of this study was to evaluate the clinical value and the prognostic power of exhaled ferritin and exhaled SOD in patients with lung cancer., Material and Methods: Forty patients with nonsmall cell lung cancer (NSCLC) and 15 controls were enrolled in the study. All subjects under study underwent EBC collection and analysis of ferritin and SOD. A total of 36 patients were either given a follow-up of at least 25.5 months or followed up until death., Results: Exhaled ferritin and SOD resulted as being higher in NSCLC than in controls and as being influenced by the stage of cancer. A pronounced survival difference was found in the presence of exhaled ferritin 300 ng/mL and exhaled SOD > 13.5 U/μL., Conclusions: In conclusion, although the results need to be confirmed on a larger and homogeneous population, we hypothesized that the notion of using the measurement of ferritin and SOD in the EBC could, if deemed feasible, have clinical implications in the monitoring of lung cancer and as an outcome predictor., (© 2011 The Authors. European Journal of Clinical Investigation © 2011 Stichting European Society for Clinical Investigation Journal Foundation.)

Induced sputum is recognized as being of increasing importance for the diagnosis and monitoring of chronic inflammatory lung diseases. The main purpose of this study is to provide a valid approach to better fractionate and characterize the still under-estimated low-molecular weight proteome of induced sputum by using mesoporous silica beads (MSBs) SPE coupled to MALDI-TOF MS. Sputum peptides were captured from both derivatized and non-derivatized MSBs and then profiled by MALDI-TOF MS. Depending on the chemical groups present on the mesoporous surface, complex peptide mixtures were extracted from induced sputum and converted into reproducible MALDI profiles. The number of peaks detected as a function of S/N was evaluated for each mesoporous surface. More than 400 peaks with an S/N>5 were obtained in comparison to 200 peaks detected without MSBs. Additionally, as a proof-of-principle, we investigated the ability of this platform to discriminate between the "sputome" of patients with asthma and chronic obstructive pulmonary disease, and between these groups and those of healthy control subjects. Six m/z peaks emerged as potential diagnostic peptidic patterns able to differentiate these inflammatory airway diseases in the sputome range. Human α-defensins (human neutrophil peptide (HNP)1, HNP2, HNP3) and three C-terminal amidated peptides, one of which is phosphorylated on serine, were identified by MALDI-TOF/TOF MS. These findings may contribute to defining a high-throughput screening MS-based platform for monitoring key peptidic-biomarkers for inflammatory and chronic respiratory diseases in induced sputum samples., (Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background: Recent advances in lung cancer biology presuppose its inflammatory origin. In this regard, LTB-4 and IL-8 are recognized to play a crucial role in neutrophil recruitment into airways during lung cancer.Notwithstanding the intriguing hypothesis, the exact role of neutrophilic inflammation in tumour biology remains complex and not completely known.The aim of this study was to give our contribution in this field by investigating LTB-4 and IL-8 in the breath condensate of NSCLC patients and verifying their role in cancer development and progression., Method: We enrolled 50 NSCLC patients and 35 controls. LTB-4 and IL-8 concentrations were measured in the breath condensate and the blood of all the subjects under study using EIA kits. Thirty NSCLC patients and ten controls underwent induced sputum collection and analysis., Results: LTB-4 and IL-8 resulted higher in breath condensate and the blood of NSCLC patients compared to controls. Significantly higher concentrations were found as the cancer stages progressed. A positive correlation was observed between exhaled IL-8 and LTB-4 and the percentage of neutrophils in the induced sputum., Conclusion: The high concentrations of exhaled LTB-4 and IL-8 showed the presence of a neutrophilic inflammation in the airways of NSCLC patients and gave a further support to the inflammatory signalling in lung cancer. These exhaled proteins could represent a suitable non-invasive marker in the diagnosis and monitoring of lung cancer.

Objectives: Dyspnea perception in asthmatics differs between subjects. Poor perception is usually associated with increased risk of asthma attack/exacerbation. The advanced stage of the disease and the presence of eosinophilic airways inflammation have been recently recognized as being responsible for poor dyspnea perception. However, few studies are available on this topic., Design: The aim of this study was to analyse the influence of inflammatory pattern, age and affective status on dyspnea perception in asthmatic subjects., Subjects and Interventions: Seventy-one consecutive asthmatic patients were recruited and underwent induced sputum, exhaled NO measurement and breath condensate collection. Perception of dyspnea was evaluated as a BORG-VAS/FEV(1) slope before and after the broncho-reversibility test and correlated with the stage of asthma, inflammatory markers, age and depression scale., Results: Dyspnea perception decreases with the worsening of asthma, with the advance of age and of depression status. Furthermore, airways inflammation plays a key role in the decline of dyspnea perception as proved by the negative correlation observed between inflammatory cells in sputum, exhaled pH and NO and BORG-VAS/FEV(1) slope., Conclusions: The results of our study suggested that airways inflammation, depression status, advance age and severity of asthma influence dyspnea perception and suggest a straight control to identify and better manage poor preceptor asthmatics., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Background: Obstructive sleep apnea syndrome (OSAS) and chronic obstructive pulmonary disease (COPD) are two diseases that often coexist within an individual. This coexistence is known as Overlap Syndrome (OS). Both diseases are characterized by local and systemic inflammations, but no studies to date have investigated local airway inflammation in patients suffering from Overlap Syndrome., Methods: We performed a Berlin Questionnaire to evaluate the presence of the principal OSAS symptoms, a pulmonary function test, and then a nocturnal oximetry and polysomnography in 72 patients that were divided into five groups: OS (n = 18), COPD (n = 15), OSAS (n = 16), 12 obese without OSAS or COPD, and one control group of 11 normal subjects. All patients underwent sputum induction and the analysis of cell patterns were evaluated in all groups. The relationship with the degree of obesity, airway obstruction and OSAS severity was also evaluated., Results: The percentage of neutrophils in induced sputum was higher in OS (74.33% ± 14.8), COPD (63.33% ± 13.22) and OSAS (60.69% ± 17.6) subjects compared with control groups of obese (43.5% ± 17.49) and normal weight (32.04% ± 12.26). No difference was found among Overlap, COPD, and OSAS patients (p = 0.56). A negative correlation was found between PaO(2) and percentage of airway neutrophils (r = -0.29, p < 0.05); similarly, no correlations arose between BMI, FEV(1) or ODI., Conclusion: Patients suffering from Overlap Syndrome present a high percentage of neutrophils in induced sputum like patients affected by COPD or OSAS alone. Our result suggests that airway inflammations is always involved in all of these diseases, even though probably sustained by different mechanisms., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

The recurrent hypoxic stress that characterizes obstructive sleep apnea (OSA) seems to play a role in the increased adherence of neutrophils to endothelial cells as well as in the resulting migration of the former to the inflamed area. Intercellular adhesion molecule 1 (ICAM-1) and interleukin (IL)-8 are markers widely used in OSA studies to investigate inflammation. The aim of this study was to measure ICAM-1 and IL-8 levels in the breath condensate and in the plasma and inflammatory cells in the induced sputum of 12 obese OSA (OO) patients, 10 nonobese OSA (NOO) patients, 10 obese non-OSA (ONO) subjects, and 8 healthy subjects (HS) using a specific enzyme immunoassay (EIA) kit. A significant increase in both plasma and exhaled IL-8 and ICAM concentrations and percentage neutrophils was observed in the induced sputum of obese OSA patients, non-obese OSA patients, and obese non-OSA subjects compared with healthy subjects. However, although these inflammatory markers were found to follow an upward trend in obese OSA patients no difference was observed in both either non-obese OSA patients and obese non-OSA subjects. Finally, a significant positive correlation was found to occur among IL-8, ICAM-1, and sputum neutrophils, as well as across the apnea-hypopnoea index (AHI), TST 90%, body mass index (BMI), and neck circumference. The data obtained confirm the occurrence of an ICAM- and IL-8-mediated neutrophilic airway inflammation in both OSA and obese patients. The degree of inflammation, which seems to worsen in cases of comorbidity (OSA and obesity), is likely to be responsible for the increased risk of developing cardiovascular events observed in these subjects, and therefore, it deserves to be elucidated even more.

Unlabelled: One of the most current intriguing hypotheses on lung cancerogenesis envisages a role for inflammation as a possible trigger of both epithelial-mesenchymal transition and cancer development. Cigarette smoke has been suggested to be the main factor underlying the inflammation of the airways described in lung cancer patients. Cycloxygenase and survivin, a COX-2 dependent factor of apoptosis resistance, seem to play a key role in this regard., Purpose: The aim of this study was to study COX-2 and survivin in the airways of lung cancer patients and in those of a group of smokers in a view to increasing our understanding of the link between smoking, airway inflammation and lung cancer., Patients and Methods: 70 NSCLC patients (28 smokers, 26 ex-smokers and 16 non-smokers) and 30 healthy subjects (20 smokers and 10 non-smokers) were enrolled in the study. Both COX-2 and survivin concentrations were measured in the exhaled breath condensates of all the subjects under study using EIA kits., Results: Higher levels of exhaled survivin and COX-2 were found in NSCLC patients compared to healthy smokers and non-smokers. These levels were observed to be significantly elevated in smokers (patients with lung cancer and healthy) and ex-smokers compared to non-smokers and exhibited a positive correlation with the number of cigarettes smoked expressed as pack/year. A correlation was also found between exhaled COX-2 and survivin and the progression of cancer., Conclusions: We support the hypothesis that cigarette smoke be strongly connected to the inflammation of the airways observed in lung cancer patients. On the basis of the results obtained the use of exhaled breath condensate COX-2 and survivin levels could be suggested as two potential markers within an early non-invasive screening of populations of smokers at risk of lung cancer.

Unlabelled: Our research group has recently been able to demonstrate and validate the possibility of studying of 3p microsatellite alterations (MAs) in the DNA extracted from the exhaled breath condensate (EBC) of healthy smokers and of subjects with non-small cell lung cancer (NSCLC). In light of the interest that has recently been aroused in the novel molecular staging protocol of lung cancer, the evaluation of the prognostic power of the genetic alterations involved in lung cancerogenesis, including 3p microsatellite alterations could be of clinical interest., Purpose: The aim of this study was to investigate the outcome predictive power of exhaled 3p microsatellite alterations in lung cancer patients., Patients and Methods: Seventy-one NSCLC patients were enrolled in the study. All the subjects under study had already undergone a 3p microsatellite analysis of their EBC. A total of 56 patients were either given a follow-up of at least 102 weeks, or were followed up until death., Results: The number of 3p microsatellite alterations found in the exhaled breath condensate DNA exhibits a remarkable correlation with patients' survival. D3S2338 and D3S1289 account for the microsatellites with the highest positive prognostic power; loss of heterozygosis (LOH) D3S1289 has a negative prognostic value in adenocarcinoma while microsatellite instability (MI) and LOH D3S2338 influence survival in squamous cell carcinoma; and, independently of NSCLC stage, D3S1289 is associated with poor prognosis., Conclusions: In conclusion, 3p MAs in the DNA of exhaled breath condensate is strongly associated with NSCLC patients' survival. Our results suggest that it is possible to use the study of EBC MAs as an outcome predictor for lung cancer patients.

One recent line of cancer research shows increasing interest for biological factor such as IL-2, TNF-alpha, and leptin, which have been found to participate in the development and progression of non-small cell lung cancer (NSCLC). The aim of this study was to measure IL-2, TNF-alpha, and leptin concentrations in the airways and in the systemic circle of patients with NSCLC, investigating the role of these factors in the lung tumors. We enrolled 32 patients (17 men, 71 +/- 7 years) with a histological diagnosis of NSCLC and 20 healthy ex-smoker controls, negative for computed tomography of the chest (14 men, 69 +/- 8 years). IL-2, TNF-alpha, and leptin levels were measured in the serum, the urine, the bronchoalveolar lavage, the induced sputum, and exhaled breath condensate (EBC) of patients enrolled by means of a specific enzyme immunoassay kit. Higher concentrations of IL-2, TNF-alpha and leptin were found in NSCLC patients than in controls (p < 0.0001). A statistically significant increase of IL-2, TNF-alpha, and leptin concentrations was observed in patients from stage I to stage III of NSCLC. These findings suggest that IL-2, TNF-alpha, and the leptin play an important role in the cancerogenesis of NSCLC. Their measure in the EBC could be proposed as noninvasive markers for an early detection of NSCLC and in the follow-up of this tumor.