Exhaled Breath Condensate Collection Research Articles

Exhaled Breath Condensate-A Non-Invasive Approach for Diagnostic Methods in Asthma.

The pathophysiology of asthma has been intensively studied, but its underlying mechanisms such as airway inflammation, control of airway tone, and bronchial reactivity are still not completely explained. There is an urgent need to implement novel, non-invasive diagnostic tools that can help to investigate local airway inflammation and connect the molecular pathways with the broad spectrum of clinical manifestations of asthma. The new biomarkers of different asthma endotypes could be used to confirm diagnosis, predict asthma exacerbations, or evaluate treatment response. In this paper, we briefly describe the characteristics of exhaled breath condensate (EBC) that is considered to be an interesting source of biomarkers of lung disorders. We look at the composition of EBC, some aspects of the collection procedure, the proposed biomarkers for asthma, and its clinical implications. We also indicate the limitations of the method and potential strategies to standardize the procedure of EBC collection and analytical methods.

Z-alpha1-antitrypsin polymers and small airways disease: a new paradigm in alfa-1 anti-trypsin deficiency-related COPD development?

The presence of Alpha1-Antitrypsin (AAT) polymers, known to promote a sustained pro-inflammatory activity, has been previously demonstrated in bronchial biopsies of subjects with Z-AAT deficiency (AATD) suggesting a possible role in the development of COPD through a small airway disease impairment. The study aimed to assess the presence of small airways dysfunction and the potential correlation with the presence of Z-AAT polymers obtained by Exhaled Breath Condensate (EBC) collection in PiZZ subjects, as compared with matched healthy PiMM subjects. We enrolled 19 asymptomatic, never smoker subjects: 9 PiZZ and 10 PiMM as controls, without obstructive ventilatory defect (i.e., normal FEV1/VC% ratio). All subjects underwent complete pulmonary function tests (PFT). EBC was collected in all subjects. ELISA test was applied to search for Z-AAT polymers. The PiZZ subjects showed normal lung volumes and DLCO values. However, in comparison with PiMM subjects, the single breath test N2 wash-out revealed significant differences regarding the phase III slope (1.45±0.35 N2/L vs. 0.96±0.40 N2/L) (p<0.02) in the PiZZ subjects, while the closing volume/vital capacity ratio (14.3±4.5 % vs. 11.3±6.3 %) was not significantly increased. The ELISA test detected the presence of Z-AAT polymers in 44% of PiZZ patients. Asymptomatic, never smoker PiZZ subjects with normal spirometry and lung diffusion capacity showed airways impairment when compared to PiMM subjects. Although Z-AAT polymers were found only in 44% of PiZZ subjects, these findings suggest the possibility that chronic bronchiolitis can develop as a result of the long-term pro-inflammatory activity of Z-AAT polymers in subjects with Z-related AATD.

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.

Isoprostane-8 in the exhaled breath condensate: Could it represent a noninvasive strategic tool for primary ciliary dyskinesia diagnosis and management?

Background and Objectives:Exhaled breath condensate (EBC) represents a potential diagnostic tool for Primary Ciliary Diskinesia (PCD). An increased oxidative stress is present in the airways of children affected and many neutrophil chemoattractants and markers of oxidative stress can be involved. The aim of the study is to evaluate leukotriene B4 (LTB-4), interleukin 8 (IL-8), 8-isoprostane (8-IP) concentration in PCD subjects, investigating their potential role as non-invasive markers of inflammation for the diagnosis and management of PCD.Methods:Cross-sectional study. 43 patients were enrolled in the study and divided in two groups: PCD (27) and healthy subjects (16). Physical examination, lung function test, nFeNO measurement and EBC collection were performed in all subjects.Results:PCD subjects showed an EBC 8-IP concentration significantly higher than the control group (median value: 11.9 pg/ml; IQR, 5.5–24.0 vs. median, 6.7 pg/ml; IQR, 2.5–11.3, p-value of Wilcoxon rank-sum test 0.0436). LTB4 EBC concentration did not differ between the two group (median, 4.3 pg/ml; IQR, 3.0–8.8 vs. 7.5 pg/ml; IQR, 3.0–9.5; P=0.4901). No significant correlation was found between FEV1 and EBC 8-IP (r=-0.10, P=0.6314) or LTB4 concentration (r=0.03, P=0.8888) in PCD subjects. No significant correlation was found between nFeNO and EBC 8-IP (r=-0.31, P=0.1385) or LTB4 (r=0.04, P=0.8565) in PCD subjects.Conclusions:EBC 8-IP levels are significantly increased in PCD subjects, highlighting the role of oxidative stress in airway inflammation. It could have a potential role as a non-invasive marker of inflammation for the diagnosis and management of PCD, although a therapeutic application of this evidence seems far.

Non-Invasive Infections Diagnosis in Lung Transplant Recipients in Exhaled Breath Condensate

Purpose The purpose of this research was to identify diagnostic volatile organic compounds (VOCs) in exhaled breath condensate (EBC) from lung transplant recipients (LTR) colonized or infected by Aspergillus spp., Staphylococcus aureus and Pseudomonas aeruginosa, in order to develop a cheap, accurate and quick non-invasive diagnostic test. To this end we performed metabolomic analysis in EBC using gas chromatography-time-of-flight mass spectrometry (GC-QTOF) in the high-resolution mode. Methods Between January 2018-June 2019, 40 LTR were recruited in the Lung Transplant Unit, from whom 71 EBC and respiratory samples (sputum, bronchoalveolar lavage or oropharyngeal swabs) were obtained. Bacteria and fungi identification were performed according to routine laboratory standards for respiratory samples. The ECOScreen2 device was used for EBC collection at -20 °C during 15 minutes, and samples were stored at −80°C in 200 μL and 500 μL aliquots until analysis. EBC samples from patients colonized either with P. aeruginosa, S. aureus, Aspergillus spp. or none of the above (controls) where selected for headspace Solid Phase Micro Extraction (SPME) with DVBP fibres in order to concentrate VOCs before their injection in the Agilent GCMS 7200 Accurate Mass Q-TOF equipment for the metabolomic analysis. GC-QTOF untargeted analysis (deconvolution, alignment, and recursive analysis) was performed with the MassHunter and Mass Profiler Software, and statistical analysis with R software. Results We selected 27 samples for SPME-GC/QTOF analysis: nine controls, seven from patients colonized by Aspergillus spp., six with P. aeruginosa, three with S. aureus and two with both P. aeruginosa and S. aureus. The PLS-DA analysis clearly differentiated patients colonized with S. aureus or P. aeuruginosa from those who were not, with a very little overlap in patients colonized with A. fumigatus from the controls. Volcano plot analysis found two possible biomarkers for Aspergillus spp. detection, six for S. aureus and none for P. aeruginosa, may be because of the small n in our study. Conclusion SPME-GC/QTOF in EBC is a promising tool for non-invasive diagnosis of fungal and bacterial infections in LTR. However, the presumptive identification of these biomarkers needs to be confirmed with targeted metabolomic studies and their sensitivity and specificity should be studied in future research.

Exhaled breath condensate in acute pulmonary embolism; a porcine study of effect of condensing temperature and feasibility of protein analysis by mass spectrometry

The search for diagnostic biomarkers for pulmonary embolism (PE) has mainly been focused on blood samples. Exhaled breath condensate (EBC) is a possible source for biomarkers specific for chronic lung diseases and cancer, yet no previous studies have investigated the potential of EBC for diagnosis of PE. The protein content in the EBC is very low, and efficient condensing of the EBC is important in order to obtain high quality samples for protein analysis. We investigated if advanced proteomic techniques in a porcine model of acute intermediate-high-risk PE was feasible using two different condensing temperatures for EBC collection. Seven pigs were anaesthetized and intubated. EBC was collected one hour after intubation. Two autologous emboli were induced through the right external jugular vein. Two hours after the emboli were administered, EBC was collected again. Condensing temperature was either −21 °C or −80 °C. Nano liquid chromatography-tandem mass spectrometry (nLC-MS/MS) was used to identify and quantify proteins of the EBC. A condensing temperature of −80 °C significantly increased the EBC volume compared with −21 °C (1.78 ± 0.25 ml vs 0.71 ± 0.12 ml) while the protein concentration in the EBC was unaltered. The mean protein concentration in the EBCs was 5.85 ± 0.93 µg ml−1, unaltered after PE. In total, 254 proteins were identified in the EBCs. Identified proteins included proteins of the cytoplasm, nucleus, plasma membrane and extracellular region. The protein composition did not differ according to condensing temperature. The EBC from pigs with acute intermediate-high-risk PE contained sufficient amounts of protein for analysis by nLC-MS/MS. The proteins were from relevant cellular compartments, indicating that EBC is a possible source for biomarkers for acute PE.

Nucleic acid detection and quantitative analysis of influenza virus using exhaled breath condensate

Exhaled breath condensate (EBC) is increasingly being used as a non-invasive method for disease diagnosis and environmental exposure assessment. We previously detected the nucleic acids of bacterial pathogens in EBC. Influenza viruses can be transmitted through aerosols during coughing and exhaling. Existing detection methods for influenza have various limitations. The EBC collection method is convenient, non-invasive, and reduces the risk of exposure. We investigated the detection of influenza virus in EBC using a sensitive nucleic acid testing method and performed quantitative analysis to evaluate the present and content of influenza virus in the breath. We evaluated 30 patients with respiratory tract infection during the 2019 influenza season. The clinical data and samples of nasal swabs were collected for rapid influenza diagnostic (antigen) tests. Pharyngeal swab and EBC samples were used for influenza virus nucleic acid detection. Each EBC sample was assessed twice as well as at one-month follow-up of the patients. The nucleic acid test in the EBC of 30 cases revealed 20 and two cases of influenza A and B, respectively, giving a detection rate of 73.3%. The rapid influenza diagnostic (antigen) tests revealed four and 12 cases of influenza A and B, respectively, with a detection rate of 53.3%. All pharyngeal swab samples evaluated by the nucleic acid test were influenza-positive; 12 cases were positive for both influenza A and B and 18 cases were positive for influenza B alone. The influenza viral load in the EBC was 103–107 copies ml−1. Among the 16 patients followed-up after 1 month, 4 were positive (25%) in EBC samples and 10 were positive (62.5%) in pharyngeal swab samples. It was preliminary exploration that influenza virus could be detected in EBC. EBC is one of the sample types that would be used for molecular diagnosis of influenza.

The influence of active and passive air humidification on exhaled breath condensate volume

Exhaled breath condensate (EBC) is safely collected in mechanically ventilated (MV) patients, but there are no guidelines regarding humidification of inhaled air during EBC collection. We investigated the influence of active and passive air humidification on EBC volumes obtained from MV patients.We collected 29 EBC samples from 21 critically ill MV patients with one condition of active humidification and four different conditions of non-humidification; 19 samples from 19 surgical MV patients with passive humidification and two samples from artificial lungs MV with active humidification. The main outcome was the obtained EBC volume per 100 L exhaled air.When collected with different conditions of non-humidification, mean [95% CI] EBC volumes did not differ significantly (1.35 [1.23; 1.46] versus 1.16 [1.05; 1.28] versus 1.27 [1.13; 1.41] versus 1.17 [1.00; 1.33] mL/100 L, p=0.114). EBC volumes were higher with active humidification than with non-humidification (2.05 [1.91; 2.19] versus 1.25 [1.17; 1.32] mL/100 L, p<0.001). The volume difference between these corresponded to the EBC volume obtained from artificial lungs (0.81 [0.62; 0.99] versus 0.89 mL/100 L, p=0.287). EBC volumes were lower for surgical MV patients with passive humidification compared to critically ill MV patients with non-humidification (0.55 [0.47; 0.63] versus 1.25 [1.17; 1.32] mL/100 L, p<0.001).While active humidification increases EBC volumes, passive humidification decreases EBC volumes and possibly influences EBC composition by other mechanisms. We propose that EBC should be collected from MV patients without air humidification to improve reproducibility and comparability across studies, and that humidification conditions should always be reported.

SMALL AIRWAYS DISEASE AND Z-ALPHA1-ANTITRYPSIN POLYMERS: IS THERE A CORRELATION?

SESSION TITLE: Late-breaking Abstract Posters SESSION TYPE: Original Investigation Posters PRESENTED ON: October 18-21, 2020 PURPOSE: The aim of the study was to assess the presence of small airways dysfunction and the potential correlation with the presence of Z-AAT polymers obtained by Exhaled Breath Condensate (EBC) collection in PiZZ subjects, comparing with matched healthy PiMM subjects. METHODS: We enrolled 17 asymptomatic non-smoker subjects: 9 PiZZ and 8 PiMM as controls, without obstructive ventilatory defect (i.e.: normal FEV1/VC ratio). All subjects underwent complete pulmonary function tests (PFT). EBC was collected in PiZZ patients. ELISA test was applied to search for Z-AAT polymers. RESULTS: The PiZZ subjects showed normal lung volumes and DLCO values. However, In comparison with PiMM subjects, the single breath test N2 wash-out revealed significant differences regarding the phase III slope (1.5±0.4 N2/L vs 1.1±0.3 N2/L) and the closing volume/vital capacity ratio (14.3±4.5 % vs 9.5±5.3 %) (p<0.05) in the PiZZ subjects. The ELISA test detected the presence of Z-AAT polymers in 4 PiZZ patients. No correlations were found between polymers levels and any functional respiratory parameter. CONCLUSIONS: PiZZ subjects showed small airways dysfunction when compared to PiMM subjects. Surprisingly, Z-AAT polymers were found only in 4 PiZZ subjects; this is probably due to the sample methodology. New studies are needed to evaluate the correlation between bronchial Z-AAT polymers and small airway impairment in subjects with Z-AAT deficiency. CLINICAL IMPLICATIONS: The presence of Alpha1-Antitrypsin (AAT) polymers that are pro-inflammatory has been previously demonstrated in bronchial biopsies of subjects with Z-AAT deficiency (Respir Res, 2014 Sep 14;15:112), suggesting a possible role in the development of COPD. DISCLOSURES: No relevant relationships by Marianna Arici, source=Web Response No relevant relationships by Jordan Franz Giordani, source=Web Response No relevant relationships by Guido Levi, source=Web Response No relevant relationships by Laura Pini, source=Web Response No relevant relationships by Chiara Rocchetti, source=Web Response No relevant relationships by Claudio Tantucci, source=Web Response No relevant relationships by Laura Tiberio, source=Web Response

Investigating the relationship between breath aerosol size and exhaled breath condensate (EBC) metabolomic content

Exhaled breath aerosols contain valuable metabolomic content due to gas exchange with blood at the alveolar capillary interface in the lung. Passive and selective filtering of these aerosols and droplets may reduce the amount of saliva contaminants and serve as an aid to enhance targeted metabolomic content when sampled in exhaled breath condensate (EBC). It is currently unknown if breath aerosol size distribution affects the types or abundances of metabolites sampled through EBC. This pilot study uses a previously described hand-held human breath sampler device with varying notch filter geometries to redirect the trajectory of breath aerosols based on size. Ten notch filter lengths were simulated with the device to calculate the effect of filter length on the breath aerosol size distribution and the proportion of aerosols which make their way through to an EBC collection tube. From three notch filter lengths, we investigate metabolite content of various aerosol fractions. We analyzed the non-volatile fraction of breath condensate with high performance liquid chromatography-mass spectrometry for broad metabolite coverage. We hypothesize that: (1) increasing the length of the notch filter in this device will prevent larger aerosols from reaching the collection tube thus altering the breath aerosol size distribution sampled in EBC; and (2) there is not a systematic large-scale difference in EBC metabolomic content that correlates with breath aerosol size. From simulation results, particles typically larger than 10 µm were filtered out. This indicates that a longer notch filter in this device prevents larger particles from reaching the collection tube thus altering the aerosol particle size distribution. Most compounds were commonly present in all three filter lengths tested, and we did not see strong statistical evidence of systematic metabolite differences between breath aerosol size distributions.

Swimming And Respiratory System: Impact Of Exercise On Pro-oxidants Production And Lung Function

The respiratory redox state of swimmers can be affected by the increase of minute ventilation (VE) during exercise and/or by chronic exposure to chlorine used to sanitize the water of swimming pools. However, in indoor-pools, the high-humidity and warm-water temperature are recognized as respiratory protecting factors. The exhaled breath condensate (EBC) is a non-invasive method used to assess the pro-oxidants species (EBC) is a non-invasive method used to assess the pH and pro-oxidants species (nitrite [NO2-], hydrogen peroxide [H2O2]), while spirometry the lung function (FEV1, FEV1/FVC, FEF25-75% ). PURPOSE: To assess the impact of a high-intensity and prolonged-time exercise on the production of [H2O2]EBC, [NO2-]EBC, pHEBC and lung function in swimmers. METHODS: Longitudinal quasi-experimental study. 18 competitive swimmers (8 female) (22±2 years; 53.1±3.5 mL·kg-1·min-1 VO2-máx.) completed 3.500-m of swim in indoor pool treated by chlorine (intensity of 80.2±3.1% of HRmax.) with no exposure to respiratory irritants by 5-days previous to tests. The spirometry test and EBC collection were done at rest, 20-minutes and 24-hours post-exercise and were analyzed using one-way RM-ANOVA test by the GraphPad-Prism software (v.6.0). The p-value<0.05 was considered for differences. RESULTS: The [H2O2]EBC (0.35±0.15 vs 0.28±0.12 vs 0.20±0.10 (umol·L-1 )) and [NO2-]EBC (1.79±0.21 vs 1.37±0.15 vs 1.03±0.16 (umol·L-1 )) decreased significantly at 24-hr post-exercise. The pHEBC (p=0.23 and 0.32) and FEV1, FEV1/FVC, FEF25-75% do not changed significantly between stages. CONCLUSION: A acute high-intensity and prolonged-time swimming session decreased the pro-oxidants production with no changes in lung function in swimmers exposed chronically to exercise and respiratory irritants. More studies are necessary to identify and isolate the plausible factors involucrated in the formation of respiratory pro-oxidants during exercise in athletes chronically exposed to respiratory irritants.

Reference Ranges of 8-Isoprostane Concentrations in Exhaled Breath Condensate (EBC): A Systematic Review and Meta-Analysis.

Isoprostanes are physiopathologic mediators of oxidative stress, resulting in lipid peroxidation. 8-isoprostane seems particularly useful for measuring oxidative stress damage. However, no reference range values are available for 8-isoprosante in exhaled breath condensate (EBC) of healthy adults, enabling its meaningful interpretation as a biomarker. We conducted this systematic review and meta-analysis according to the protocol following PROSPERO (CRD42020146623). After searching and analyzing the literature, we included 86 studies. After their qualitative synthesis and risk of bias assessment, 52 studies were included in meta-analysis. The latter focused on studies using immunological analytical methods and investigated how the concentrations of 8-isoprostane differ based on gender. We found that gender had no significant effect in 8-isoprostane concentration. Among other studied factors, such as individual characteristics and factors related to EBC collection, only the device used for EBC collection significantly affected measured 8-isoprostane concentrations. However, adjustment for the factors related to EBC collection, yielded uncertainty whether this effect is due to the device itself or to the other factors. Given this uncertainty, we estimated the reference range values of 8-isoprostane stratified by gender and EBC collection device. A better standardization of EBC collection seems necessary; as well more studies using chemical analytical methods to extend this investigation.

Does exposure to inflammatory particles modify the pattern of anion in exhaled breath condensate?

Exposure to environmental and occupational particulate matter (PM) induces health effects on the cardio-pulmonary system. In addition, associations between exposure to PM and metabolic syndromes like diabetes mellitus or obesity are now emerging in the literature. Collection of exhaled breath condensate (EBC) is an appealing non-invasive technique to sample pulmonary fluids. This hypothesis-generating study aims to (1) validate an ion chromatography method allowing the robust determination of different metabolism-related molecules (lactate, formate, acetate, propionate, butyrate, pyruvate, nitrite, nitrate) in EBC; (2) apply this method to EBC samples collected from workers exposed to quartz (a known inflammatory particle), to soapstone (a less inflammatory particle than quartz), as well as to controls. A multi-compound standard solution was used to determine the linearity range, detection limit, repeatability and bias from spiked EBC. The biological samples were injected without further treatment into an ion chromatograph with a conductivity detector. RTube® were used for field collection of EBC from 11 controls, 55 workers exposed to soapstone and 12 volunteers exposed to quartz dust. The analytical method used proved to be adequate for quantifying eight anions in EBC samples. Its sub-micromolar detection limits and repeatability, combined with a very simple sample preparation, allowed an easy and fast quantification of different glycolysis or nitrosative stress metabolites. Using multivariate discriminant analysis to maximize differences between groups, we observed a different pattern of anions with a higher formate/acetate ratio in the EBC samples for quartz exposed workers compared to the two other groups. We hypothesize that a modification of the metabolic signature could be induced by exposure to inflammatory particles like quartz and might be observed in the EBC via a change in the formate/acetate ratio.

Safety and Yield of Exhaled Breath Condensate Analysis in Acutely Ill, Mechanically Ventilated Infants with RSV Bronchiolitis

Exhaled breath condensate (EBC) analysis is a rapidly growing field of research and may be a useful non-invasive technique to evaluate lung pathology. There is little information in the literature on the safety of collecting these samples in acutely ill infants needing mechanical ventilation. Viral bronchiolitis is a common lower respiratory tract infection contributing to the majority of hospitalizations in children under 2 years of age. The pathophysiology of severe bronchiolitis resulting in ventilatory support is illdefined. Our study objective was to evaluate the tolerability of collection and utility of exhaled breath condensate (EBC) in molecular evaluation of viral bronchiolitis. We obtained proteomic profile of sequential EBCs over three days of illness from two infants who were intubated and ventilated. Mass spectrometry analysis of these samples identified cytokeratin at all time-points and time-dependent increase of proteins related to intracellular adhesions, epidermal morphogenesis, cell repair and differentiation that may be used to monitor the status of epithelial barrier function and integrity of intercellular communication in the bronchiolitis lung in the future. Our pilot study highlights that EBC collection is a well- tolerated procedure to obtain meaningful longitudinal data to profile and study the molecular parameters in acutely ill bronchiolitis infants.

Capillary electrophoretic analysis of ionic content in exhaled breath condensate and pH monitoring as a non-invasive method in gastroesophageal reflux disease diagnostics

In this study, the ionic profile and pH of exhaled breath condensate (EBC) in a group of patients with acid and weakly acid reflux and no-reflux controls were compared. A portable sampler was used for non-invasive EBC collection from five exhalations. The ionic profile (anions, cations, organic acids) and pH of the collected EBC samples were measured by capillary electrophoresis with contactless conductivity detection and a pH microelectrode, respectively. Several ions were elevated in the patient groups. Sodium cation was elevated in weakly acid reflux (significance level p < 0.01) and acid reflux (p < 0.05) compared to no-reflux controls. Butyrate and propionate were elevated in both acid reflux and weakly acid reflux compared to no-reflux controls (butyrate: p < 0.01, propionate: p < 0.05). The median values of pH (after de-aeration with N2) were also significantly higher (p < 0.01) in groups with acid reflux and weakly acid reflux than in the control group with no reflux. The ionic analysis and simultaneous pH measurement offer a simple, cheap, fast, and non-invasive approach in gastroesophageal reflux disease diagnostics.

Detecting opioid metabolites in exhaled breath condensate (EBC)

Exhaled breath condensate (EBC) collection provides a promising matrix for bioanalysis of endogenous biomarkers of health and also for exogenous compounds like drugs. There is little information regarding drugs and their metabolites contained in breath, as well as their pharmacokinetics. In this present work, we use a simple and non-invasive technique to collect EBC from chronic pain patients using different analgesic opioid drugs to manage pain. Six patients received continuous infusion of morphine and hydromorphone intravenously (IV), together with other analgesic drugs (IV and orally). Repeated sampling of serum and EBC was done at two time points separated by 90 min. The EBC was collected using a glass tube surrounded by dry ice, and an ethanol solvent wash of the glass was performed after EBC extraction to retrieve the apolar compounds stuck to the glass surface. All samples were analyzed with liquid chromatography coupled to mass spectrometry (LC-MS/MS) to identify possible metabolites present in the sample, and to quantify the drugs being used. Several metabolites, such as normorphine (norM), norhydromorphone (norHM) and dihydromorphone (diHM) were detected in both fractions, while hydromorphone 3-glucuronide (HM 3G) was only detected in the solvent rinse fraction. Results were correlated to explain the pharmacokinetics of the main drugs administered. This pilot study presented promising correlations between drug concentrations in blood and breath at different time points for norM, norHM and HM 3G.

A sampler prototype for the simultaneous collection of exhaled air and breath condensate.

Exhaled air and breath condensate contain a large number of health biomarkers, such as volatile and semi-volatile organic compounds, proteins and lipids. Nowadays, the collection of breath samples is carried out by commercial or lab-made sampling systems that collect only one type of sample (e.g. gaseous or condensate phase), thus limiting the diagnostic capability of breath tests. This work presents a portable prototype optimized for the simultaneous collection of gaseous exhaled breath and exhaled breath condensate within five minutes. The system is fully portable and has a total weight of about 1 Kg. An illustrative determination of ethanol, isoprene, acetone, isopropyl alcohol, 1-propanol, 2-butanone, 2-pentanone, toluene and xylenes in breath, and cortisol and 8-iso-prostaglandin F2α in breath condensate is discussed.

CfDNA in exhaled breath condensate (EBC) and contamination by ambient air: toward volatile biopsies

Exhaled breath is a source of volatile and nonvolatile biomarkers in the body that can be accessed non-invasively and used for monitoring. The collection of lung secretions by conventional methods such as bronchoalveolar lavage, induced sputum collection, and core biopsies is limited by the invasive nature of these methods. Non-invasive collection of exhaled breath condensate (EBC) provides fluid samples that are representative of airway lining fluids. Various volatile and nonvolatile biomarkers can be detected in volatile condensates, such as H2O2, nitric oxide, lipid mediators, cytokines, chemokines, DNA, and microRNAs. Studies have examined cell-free DNA (cfDNA) in plasma samples from non-small-cell lung cancer patients, offering to new insights and fostering development of the liquid biopsy. However, few studies have examined cfDNA in EBC samples. This study examined whether EBC is an appropriate source of cfDNA using housekeeping-gene-specific primer probes and quantitative real-time polymerase chain reaction in healthy subjects. Ambient (room) air is contaminated with DNA, so caution is needed. Preliminary studies indicated that volatile biopsies are becoming an important diagnostic tool in lung cancer.

Exhaled Breath Condensate in Childhood Asthma: A Review and Current Perspective.

Exhaled breath condensate (EBC) was introduced more than two decades ago as a novel, non-invasive tool to assess airway inflammation. This review summarizes the latest literature on the various markers in EBC to predict asthma in children. Despite many recommendations and two comprehensive Task Force reports, there is still large heterogeneity in published data. The biggest issue remains a lack of standardization regarding EBC collection, preservation, processing, and analysis. As a result, published studies show mixed or conflicting results, questioning the reproducibility of findings. A joint, multicenter research study is urgently needed to address the necessary methodological standardization.

Looking for Airways Periostin in Severe Asthma: Could It Be Useful for Clustering Type 2 Endotype?

Severe asthma is heterogeneous clinically and biologically and is often difficult to control. In particular, the type 2 (T2) immunity endotype of severe asthma is gaining increasing interest because it is susceptible to newly developed biologic treatments that can transform the quality of life of these patients. The aim of this study was to analyze periostin concentrations in the airways of patients with severe asthma, evaluating its role in clustering the T2 endotype. We enrolled 40 consecutive patients with severe asthma (T2 endotype: n= 25; non-T2 endotype: n= 15), 21 patients with mild to moderate asthma, and 15 healthy control subjects. All subjects enrolled underwent exhaled breath condensate (EBC) and sputum collection, eosinophil count in blood, fractional exhaled nitric oxide, and IgE measurement. Periostin was assessed by an enzyme-linked immunosorbent assay kit on EBC and induced sputum (IS) supernatant. We were able to detect higher periostin levels in the EBC (0.75 ± 0.46 vs0.70 ± 0.19 vs0.11 ± 0.05ng/mL, P< .05 and P< .01) and in IS (0.55 ± 0.23 vs0.31 ± 0.13 vs0.16 ± 0.120ng/mL, P< .05 and P< .01) of patients with severe asthma compared with patients with mild to moderate asthma and healthy control subjects, respectively. We further found an increase of periostin levels in both samples in T2 endotype compared with non-T2 endotype (EBC: 0.88 ± 0.46 vs0.52 ± 0.46ng/mL; IS: 0.69 ± 0.19 vs0.39 ± 0.16ng/mL; P<.05) and a correlation between periostin levels in EBC and sputum. We found that periostin is measurable in the airways and increased in patients with severe asthma, especially in those from the T2 endotype. Unlike serum periostin, which may be derived from several sources outside the lung, airways periostin is a useful marker of severe eosinophilic asthma and may help to phenotype patients that will respond to the biologic agents.