The Potential Role of Airways Periostin in the Clinical Practice of Patients Affected by Idiopathic Pulmonary Fibrosis.

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.

Assessment of Induced Sputum Cellularity in COPD Patients Belonging to Two Different Classes of Air Pollution Exposure / Estudio de la celularidad en el esputo inducido de pacientes con EPOC expuestos a dos niveles diferentes de contaminación atmosférica

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

INTRODUCCIÓN: Varios estudios han demostrado con anterioridad que la exposición a la contaminación atmosférica a largo plazo provoca una inflamación de la vía aérea que se ve reflejada en un aumento de neutrófilos en el esputo. OBJETIVO Y MÉTODOS: El objetivo de nuestro estudio fue estimar el grado de inflamación de la vía aérea a través del esputo inducido en un grupo de 15 pacientes con EPOC bien caracterizada sin exposición por motivos profesionales y residentes en áreas urbanas con un nivel alto de contaminación atmosférica, comparados con un grupo control de 13 individuos con EPOC que vivían en zonas rurales con un nivel de contaminación atmosférica bajo. A todos los pacientes se les sometió a una espirometría y a una inducción del esputo. RESULTADOS: Se encontró un aumento estadísticamente significativo en el porcentaje de recuento celular de neutrófilos de los residentes en áreas urbanas en comparación con aquellos que vivían en zonas rurales (89,1 frente a 79,0%, p < 0,05). CONCLUSIONES: En conclusión, demostramos que los pacientes con EPOC que no estaban expuestos debido a su profesión y residentes en áreas urbanas con alta contaminación atmosférica presentaban mayor inflamación de la vía aérea en cuanto al número de neutrófilos en esputo en comparación con una población de características muy similares residente en zonas rurales con menos contaminación atmosférica. El resultado de este estudio piloto podría ser relevante para el efecto a largo plazo de la contaminación atmosférica exterior en pacientes especialmente vulnerables como pueden ser aquellos con EPOC

Assessment of Induced Sputum Cellularity in COPD Patients Belonging to Two Different Classes of Air Pollution Exposure.

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.

Expression profiling of miRNA-145 and miRNA-338 in serum and sputum of patients with COPD, asthma, and asthma-COPD overlap syndrome phenotype.

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.

Microparticles in sputum of COPD patients: a potential biomarker of the disease?

BACKGROUND: 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<0.05) and CD66b-MP level was correlated with worse performance index of COPD such as the Body mass index airflow Obstruction, Dyspnea, and Exercise capacity (BODE); they were negatively correlated with 6-minute walking test: 0.65 and -0.64, respectively (P<0.05). CD235ab-MPs showed a negative correlation with body mass index (R=-0.86, P<0.05), while there was a positive correlation with dyspnea index (R=0.91, P<0.05). CONCLUSION: The main finding of this study was that MPs were detected in the sputum of patients affected by COPD. The phenotype of some of them was related to the main COPD parameters. These results suggest that MPs could be implicated in the pathogenesis of COPD.

Exhaled HPV infection in lung cancer: role of MA at 3p.

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.

Aspergillus spp. colonization in exhaled breath condensate of lung cancer patients from Puglia Region of Italy.

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.

Role of vasoactive intestinal peptide in chronic obstructive pulmonary disease with pulmonary hypertension.

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.

Aging and airway inflammation.

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.

[New biomolecular methodologies in diagnosis of lung cancer]. / Attualità sui nuovi metodi biomolecolari nella diagnosi di neoplasie polmonari.

Lung cancer remains the most frequent tumour and cause of cancer death in worldwide. Unfortunately most of patients still discover their tumour in advanced stage. Lung cancer results from the occurrence of a number of genetic alterations in oncogenes and tumour suppressor genes that are potential markers either for screening procedures or for earlier detection in patients with non small-cell lung cancer (NSCLC). It was estimated that about 10 to 20 genetic events are required for lung tumorigenesis. These genetic changes are triggered by smoking and persist for many years after smoking cessation. Continuously, more sophisticated methods for the analysis of these genetic alterations involved in lung cancer become available. Several molecular alterations involved in lung cancer have been already identified in different biological samples (biopsy, BAL) that are collected with highly invasive techniques that make them poorly suitable for wider screening. Recently the DNA has been extracted from exhaled breath condensate, demonstrating the suitability of this sample for the study of genetic alterations and its potential role in screening programs of subjects at risk of lung cancer.