The Experts below are selected from a list of 4587 Experts worldwide ranked by ideXlab platform
Peter J. Barnes - One of the best experts on this subject based on the ideXlab platform.
-
nmr spectroscopy metabolomic profiling of Exhaled Breath Condensate in patients with stable and unstable cystic fibrosis
Thorax, 2012Co-Authors: Paolo Montuschi, Peter J. Barnes, Giovanni Ciabattoni, Andrew Bush, Debora Paris, Dominique Melck, Vincenzina Lucidi, Valeria Raia, Cecilia Calabrese, Andrea MottaAbstract:Background Metabolomics could provide new insights into the pathophysiology of cystic fibrosis (CF) by identifying profiles of endogenous metabolites. Objectives To investigate whether metabolomics of Exhaled Breath Condensate could discriminate between patients with unstable CF, stable CF and healthy subjects, and whether selected metabolites were responsible for between-group differences. Methods Twenty-nine patients with stable CF, 24 with unstable CF and 31 healthy subjects (age 9–24 years) participated in a cross-sectional study. Metabolomics was performed with high-resolution nuclear magnetic resonance spectroscopy. Partial least squares-discriminant analysis was used as classifier. The results were validated in a second independent study. Results Intraclass correlation coefficients for between-day and technical repeatability were 0.93 and 0.96, respectively. Bland–Altman analysis showed good within-day repeatability. Correct classification rate of CF (n=53) vs healthy subjects (n=31) was 96% (R 2 =0.84; Q 2 =0.79). Model validation with a testing sample set obtained from subjects not included in the primary analysis (23 CF and 25 healthy subjects) showed a sensitivity of 91% and a specificity of 96%. The classification rate of stable CF (n=29) vs unstable CF patients (n=24) was 95% (R 2 =0.82; Q 2 =0.78). Model external validation in 14 patients with stable CF and 16 with unstable CF showed a sensitivity of 86% and a specificity of 94%. Ethanol, acetate, 2-propanol and acetone were most discriminant between patients with CF and healthy subjects, whereas acetate, ethanol, 2-propanol and methanol were the most important metabolites for discriminating between patients with stable and unstable CF. Conclusions Nuclear magnetic resonance spectroscopy of Exhaled Breath Condensate is reproducible, discriminates patients with CF from healthy subjects and patients with unstable CF from those with stable CF, and identifies the metabolites responsible for between-group differences.
-
correlation between eicosanoids in bronchoalveolar lavage fluid and in Exhaled Breath Condensate
Disease Markers, 2011Co-Authors: Adam Antczak, Wojciech J Piotrowski, Jerzy Marczak, Maciej Ciebiada, Pawel Gorski, Peter J. BarnesAbstract:Exhaled Breath Condensate (EBC) has been increasingly used as a new and non-invasive method to study airway inflammation. In this study we have compared the concentrations of lipid mediators in EBCwith concentrations in bronchoalveolar lavage fluid (BALF).
-
Measurement of 8-isoprostane in Exhaled Breath Condensate.
Methods in molecular biology (Clifton N.J.), 2009Co-Authors: Paolo Montuschi, Peter J. Barnes, Giovanni CiabattoniAbstract:Oxidative stress is functionally involved in the pathophysiology of lung diseases including asthma and chronic obstructive pulmonary disease. 8-Isoprostane, which is derived from free radical-catalyzed peroxidation of arachidonic acid, is one of the most reliable biomarkers of oxidative stress. Exhaled Breath Condensate (EBC) is a completely noninvasive method for collecting airway secretions. We developed a specific and sensitive radioimmunoassay (RIA) that has been applied to the measurement of 8-isoprostane in EBC. This RIA for 8-isoprostane has been validated using high performance liquid chromatography. Measurement of 8-isoprostane in EBC is a useful noninvasive technique for exploring the role of oxidative stress in lung diseases. This technique might provide important insights into the understanding of the clinical pharmacology of antioxidants and might be useful for monitoring the effects of pharmacological therapy.
-
peroxynitrite elevation in Exhaled Breath Condensate of copd and its inhibition by fudosteine
Chest, 2009Co-Authors: Grace O Osoata, Peter J. Barnes, Sergei A. Kharitonov, Toyoyuki Hanazawa, Caterina Brindicci, Misako Ito, Kazuhiro ItoAbstract:Background Peroxynitrite formed by the reaction of nitric oxide and superoxide is a powerful oxidant/nitrosant. Nitrative stress is implicated in COPD pathogenesis, but PN has not been detected due to a short half-life ( Method PN was measured using oxidation of 2′,7′-dichlorofluorescein (DCDHF) in Exhaled Breath Condensate (EBC) collected in high pH and sputum cells. The PN scavenging effect was also evaluated by the same system as PN-induced bovine serum albumin (BSA) nitration. Results The mean (± SD) PN levels in EBC of COPD patients (7.9 ± 3.0 nmol/L; n = 10) were significantly higher than those of healthy volunteers (2.0 ± 1.1 nmol/L; p Conclusion This noninvasive PN measurement in EBC may be useful for monitoring airway nitrative stress in COPD. Furthermore, FDS has the potential to inhibit PN-induced events in lung by its scavenging effect.
-
Exhaled Breath Condensate methodological recommendations and unresolved questions
European Respiratory Journal, 2005Co-Authors: Ildiko Horvath, J Hunt, Peter J. BarnesAbstract:Collection of Exhaled Breath Condensate (EBC) is a noninvasive method for obtaining samples from the lungs. EBC contains large number of mediators including adenosine, ammonia, hydrogen peroxide, isoprostanes, leukotrienes, nitrogen oxides, peptides and cytokines. Concentrations of these mediators are influenced by lung diseases and modulated by therapeutic interventions. Similarly EBC pH also changes in respiratory diseases. The aim of the American Thoracic Society/European Respiratory Society Task Force on EBC was to identify the important methodological issues surrounding EBC collection and assay, to provide recommendations for the measurements and to highlight areas where further research is required. Based on the currently available evidence and the consensus of the expert panel for EBC collection, the following general recommendations were put together for oral sample collection: collect during tidal Breathing using a noseclip and a saliva trap; define cooling temperature and collection time (10 min is generally sufficient to obtain 1-2 mL of sample and well tolerated by patients); use inert material for condenser; do not use resistor and do not use filter between the subject and the condenser. These are only general recommendations and certain circumstances may dictate variation from them. Important areas for future research involve: ascertaining mechanisms and site of Exhaled Breath Condensate particle formation; determination of dilution markers; improving reproducibility; employment of EBC in longitudinal studies; and determining the utility of Exhaled Breath Condensate measures for the management of individual patients. These studies are required before recommending this technique for use in clinical practice.
Paolo Montuschi - One of the best experts on this subject based on the ideXlab platform.
-
nmr spectroscopy metabolomic profiling of Exhaled Breath Condensate in patients with stable and unstable cystic fibrosis
Thorax, 2012Co-Authors: Paolo Montuschi, Peter J. Barnes, Giovanni Ciabattoni, Andrew Bush, Debora Paris, Dominique Melck, Vincenzina Lucidi, Valeria Raia, Cecilia Calabrese, Andrea MottaAbstract:Background Metabolomics could provide new insights into the pathophysiology of cystic fibrosis (CF) by identifying profiles of endogenous metabolites. Objectives To investigate whether metabolomics of Exhaled Breath Condensate could discriminate between patients with unstable CF, stable CF and healthy subjects, and whether selected metabolites were responsible for between-group differences. Methods Twenty-nine patients with stable CF, 24 with unstable CF and 31 healthy subjects (age 9–24 years) participated in a cross-sectional study. Metabolomics was performed with high-resolution nuclear magnetic resonance spectroscopy. Partial least squares-discriminant analysis was used as classifier. The results were validated in a second independent study. Results Intraclass correlation coefficients for between-day and technical repeatability were 0.93 and 0.96, respectively. Bland–Altman analysis showed good within-day repeatability. Correct classification rate of CF (n=53) vs healthy subjects (n=31) was 96% (R 2 =0.84; Q 2 =0.79). Model validation with a testing sample set obtained from subjects not included in the primary analysis (23 CF and 25 healthy subjects) showed a sensitivity of 91% and a specificity of 96%. The classification rate of stable CF (n=29) vs unstable CF patients (n=24) was 95% (R 2 =0.82; Q 2 =0.78). Model external validation in 14 patients with stable CF and 16 with unstable CF showed a sensitivity of 86% and a specificity of 94%. Ethanol, acetate, 2-propanol and acetone were most discriminant between patients with CF and healthy subjects, whereas acetate, ethanol, 2-propanol and methanol were the most important metabolites for discriminating between patients with stable and unstable CF. Conclusions Nuclear magnetic resonance spectroscopy of Exhaled Breath Condensate is reproducible, discriminates patients with CF from healthy subjects and patients with unstable CF from those with stable CF, and identifies the metabolites responsible for between-group differences.
-
Measurement of 8-isoprostane in Exhaled Breath Condensate.
Methods in molecular biology (Clifton N.J.), 2009Co-Authors: Paolo Montuschi, Peter J. Barnes, Giovanni CiabattoniAbstract:Oxidative stress is functionally involved in the pathophysiology of lung diseases including asthma and chronic obstructive pulmonary disease. 8-Isoprostane, which is derived from free radical-catalyzed peroxidation of arachidonic acid, is one of the most reliable biomarkers of oxidative stress. Exhaled Breath Condensate (EBC) is a completely noninvasive method for collecting airway secretions. We developed a specific and sensitive radioimmunoassay (RIA) that has been applied to the measurement of 8-isoprostane in EBC. This RIA for 8-isoprostane has been validated using high performance liquid chromatography. Measurement of 8-isoprostane in EBC is a useful noninvasive technique for exploring the role of oxidative stress in lung diseases. This technique might provide important insights into the understanding of the clinical pharmacology of antioxidants and might be useful for monitoring the effects of pharmacological therapy.
-
lc ms ms analysis of leukotriene b4 and other eicosanoids in Exhaled Breath Condensate for assessing lung inflammation
Journal of Chromatography B, 2009Co-Authors: Paolo MontuschiAbstract:Abstract Leukotriene (LT) B4 is a potent inflammatory lipid mediator that has been involved in the pathophysiology of respiratory diseases including asthma. Exhaled Breath Condensate (EBC) is a non-invasive method to sample secretions from the airways. LC/MS/MS techniques for measuring LTB4 concentrations in EBC have been developed and are suitable for an accurate quantitative assessment of its concentrations in EBC. LC/MS/MS for other eicosanoids including 8-isoprostane, a marker of oxidative stress, and cysteinyl-LTs have been developed. This article, mainly focused on LTB4, presents the analytical aspects of the LC/MS/MS techniques for measuring LTB4 and 8-isoprostane in EBC, provides examples of their application to the assessment of airway inflammation in patients with asthma and other respiratory diseases, and discusses their potential utility for non-invasive monitoring of drug therapy.
-
Exhaled Breath Condensate analysis in patients with copd
Clinica Chimica Acta, 2005Co-Authors: Paolo MontuschiAbstract:Exhaled Breath Condensate (EBC) is a non-invasive method for studying the composition of airway lining fluid. EBC is mainly formed by water vapor but also contains aerosol particles in which several biomolecules including hydrogen peroxide, leukotrienes, prostaglandins, isoprostanes, nitric oxide-derived products, and hydrogen ions have been measured in healthy subjects. Some inflammatory mediators are elevated in patients with chronic obstructive pulmonary disease (COPD). Analysis of EBC has several advantages over other methods for assessing lung inflammation: it is completely non-invasive; this technique is particularly suitable for longitudinal studies; this method is potentially useful for assessing the efficacy of pharmacological therapy. Identification of selective profiles of inflammatory markers in EBC might also be of diagnostic value in patients with COPD. EBC analysis is currently more reliable for relative measures than for determining absolute levels of inflammatory mediators. The lack of standardization of the EBC analysis is currently the primary limitation of this technique making it difficult comparisons of data obtained in different laboratories. Reference analytical techniques are required to provide definitive evidence for the presence of several biomolecules in EBC and an accurate assessment of their concentrations in this biological fluid. Moreover, several methodological issues need to be addressed before this technique can be considered in the clinical management of patients with COPD. Despite important current limitations, further research in this area is warranted due to the lack of non-invasive methods for assessing lung inflammation which has a central role in the pathophysiology of COPD.
-
validation of leukotriene b4 measurements in Exhaled Breath Condensate
Inflammation Research, 2003Co-Authors: Paolo Montuschi, Salvatore Valente, G M Corbo, G. Ciappi, Chiara Mondino, Peter J. Barnes, Enzo Ragazzoni, Giovanni CiabattoniAbstract:Objective: To qualitatively validate an enzyme immunoassay to measure leukotriene B4 in Exhaled Breath Condensate. Exhaled Breath Condensate is a new non-invasive method to monitor airway inflammation.¶Subjects: Twenty-two subjects with different lung diseases attended the outpatient clinic on one occasion for Exhaled Breath Condensate collection.¶Methods: Samples were pooled together and purified by reverse-phase high-performance liquid chromatography. The fractions eluted were assayed for leukotriene B4 by enzyme immunoassay.¶Results: A single peak of leukotriene B4-like immunoreactivity co-eluting with leukotriene B4 standard (retention time: 24 min) was identified by enzyme immunoassay. Reverse phase-high performance liquid chromatography peak of leukotriene B4 was clearly separated from those of 6-trans- leukotriene B4 (retention time: 14 min) and leukotriene B5 (retention time: 18 min) for which the antiserum used in the enzyme immunoassay had the highest cross-reactivity. Leukotriene B4 recovery was 64%.¶Conclusions: This study provides evidence for the presence of leukotriene B4 in the Exhaled Breath Condensate and the specificity of the enzyme immunoassay used.
Sergei A. Kharitonov - One of the best experts on this subject based on the ideXlab platform.
-
peroxynitrite elevation in Exhaled Breath Condensate of copd and its inhibition by fudosteine
Chest, 2009Co-Authors: Grace O Osoata, Peter J. Barnes, Sergei A. Kharitonov, Toyoyuki Hanazawa, Caterina Brindicci, Misako Ito, Kazuhiro ItoAbstract:Background Peroxynitrite formed by the reaction of nitric oxide and superoxide is a powerful oxidant/nitrosant. Nitrative stress is implicated in COPD pathogenesis, but PN has not been detected due to a short half-life ( Method PN was measured using oxidation of 2′,7′-dichlorofluorescein (DCDHF) in Exhaled Breath Condensate (EBC) collected in high pH and sputum cells. The PN scavenging effect was also evaluated by the same system as PN-induced bovine serum albumin (BSA) nitration. Results The mean (± SD) PN levels in EBC of COPD patients (7.9 ± 3.0 nmol/L; n = 10) were significantly higher than those of healthy volunteers (2.0 ± 1.1 nmol/L; p Conclusion This noninvasive PN measurement in EBC may be useful for monitoring airway nitrative stress in COPD. Furthermore, FDS has the potential to inhibit PN-induced events in lung by its scavenging effect.
-
Exhaled Breath Condensate detects markers of pulmonary inflammation after cardiothoracic surgery
American Journal of Respiratory and Critical Care Medicine, 2004Co-Authors: E Moloney, Sergei A. Kharitonov, Sharon Mumby, Gregory J. Quinlan, Reka Gajdocsi, Julius H Cranshaw, Mark D GriffithsAbstract:Cardiac surgery using cardiopulmonary by-pass and, to a greater extent, lung resection, causes acute lung injury that is usually subclinical. Analysis of mediators in Exhaled Breath Condensate is a promising means of monitoring inflammation in a variety of airway diseases but the contribution of the airway lining fluid from the lower respiratory tract is uncertain. We compared the analysis of markers of lung injury in Exhaled Breath Condensate and bronchoalveolar lavage in endotracheally intubated patients before and after coronary artery bypass graft surgery with cardiopulmonary bypass and lobectomy. The neutrophil count and leukotriene B4 concentration in bronchoalveolar lavage fluid rose after coronary artery bypass graft surgery (p < 0.05), but there was no significant change in leukotriene B4, hydrogen peroxide, or hydrogen ion concentrations in Exhaled Breath Condensate. By contrast, after lobectomy, the concentration in Exhaled Breath Condensate of leukotriene B4, hydrogen peroxide and hydrogen ions rose significantly (p < 0.05). Exhaled Breath Condensate is a safe, noninvasive method of sampling the milieu of the distal lung and is sufficiently sensitive to detect markers of inflammation and oxidative stress in patients after lobectomy, but not after the milder insult associated with cardiac surgery.
-
high levels of interleukin 6 in the Exhaled Breath Condensate of patients with copd
Respiratory Medicine, 2003Co-Authors: E Bucchioni, Sergei A. Kharitonov, L Allegra, Peter J. BarnesAbstract:Abstract Background : Chronic obstructive pulmonary disease (COPD) is characterised by chronic inflammation of the respiratory tract. Methods : We investigated the presence of interleukin-6 (IL-6: a cytokine secreted by monocytes/macrophages, T cells, B cells, fibroblasts, bone marrow stromal cells, keratinocytes and endothelial cells) in the Exhaled Breath Condensate of 16 ex-smokers with moderate COPD, 12 healthy non-smokers. IL-6 was measured by means of a specific enzyme immunoassay. Results : IL-6 levels were detectable in all of the subjects, but were higher in the COPD patients (8.0±0.1pg/ml; P r =0.597; P Conclusions : The increased IL-6 levels in Exhaled Breath Condensate may reflect airway inflammation in patients with COPD.
-
increased leukotriene b4 and interleukin 6 in Exhaled Breath Condensate in cystic fibrosis
American Journal of Respiratory and Critical Care Medicine, 2003Co-Authors: Giovanna E Carpagnano, Peter J. Barnes, D M Geddes, M E Hodson, Sergei A. KharitonovAbstract:Chronic neutrophilic airway inflammation is an important feature of cystic fibrosis (CF). Noninvasive inflammatory markers may be useful in monitoring CF. Leukotriene B4 (LTB4) and interleukin (IL)-6 are inflammatory mediators that are increased in chronic neutrophilic inflammation. The aim of this study was to assess whether LTB4 and IL-6 were increased in Exhaled Breath Condensate of CF patients and whether they could be used to monitor inflammation. Twenty patients with CF (13 males, age of 28 ± 9 years) were recruited together with 15 age-matched healthy subjects (8 males, age 35 ± 7 years). LTB4 and IL-6 levels were markedly elevated in patients with acute exacerbations (28.8 ± 4.3 and 8.7 ± 0.4 pg/ml) compared with control subjects (6.8 ± 0.7 and 2.6 ± 0.1 pg/ml, p < 0.0001). We also observed a decrease of Exhaled LTB4 and IL-6 concentrations after antibiotic treatment in six patients who were followed until clinically stable (31.1 ± 4.4 and 9.5 ± 0.4 pg/ml vs. 18.8 ± 0.8 and 6.4 ± 0.2 pg/ml, respec...
-
Increased leukotrienes in Exhaled Breath Condensate in childhood asthma.
American journal of respiratory and critical care medicine, 2002Co-Authors: Zsuzsanna Csoma, Sergei A. Kharitonov, Beatrix Balint, Andrew Bush, Nicola Wilson, Peter J. BarnesAbstract:Cysteinyl leukotrienes (cys-LTs; LTC4, LTD4, and LTE4) are generated predominantly by mast cells and eosinophils and induce airway smooth muscle contraction, microvascular leakage, and mucous hypersecretion whereas leukotriene B4 (LTB4) is a potent chemoattractant of neutrophils. We measured cys-LTs and LTB4 in Exhaled Breath Condensate from children aged 7–14 years including healthy nonatopic children (n = 11) and children with mild intermittent asthma (steroid naive, n = 11), mild persistent asthma (low-dose inhaled steroid treatment, n = 13), or moderate to severe persistent asthma (high-dose inhaled steroid treatment, n = 13). Exhaled LTB4 levels were increased in patients with mild and moderate to severe persistent asthma compared with patients with mild intermittent asthma (126.0 ± 8.8 and 131.9 ± 7.1 versus 52.7 ± 3.8 pg/ml, p < 0.001 and p < 0.0001) and normal subjects (126.0 ± 8.8 and 131.9 ± 7.1 versus 47.9 ± 4.1 pg/ml, p < 0.0001). Elevated Exhaled cys-LT levels were found in patients with mild...
John F Hunt - One of the best experts on this subject based on the ideXlab platform.
-
matrix effect in Exhaled Breath Condensate interferon gamma immunoassay
Journal of Breath Research, 2008Co-Authors: Stephanie Hom, Brian K Walsh, John F HuntAbstract:Manuscripts presenting cytokine (and other protein) concentrations in Exhaled Breath Condensate (EBC) are commonly published with insufficient control experiments performed to provide confidence that the assay was accurate. Our laboratory has identified false positives in EBC with several forms of immunoassay, including ELISA and multiplex bead arrays. In this current brief investigation, we studied EBC samples from ten subjects and examined for the effect of alteration of the matrix on the assay results reported by a commercially available ELISA assay for a commonly reported EBC cytokine (interferon-gamma (IFN-γ)). The commercial immunoassay reported measurable levels of the IFN-γ signal in all samples. However, when we added to the EBC samples the proteinaceous matrix of the zero-standard from the commercial kit, we could identify no IFN-γ signal. These simple findings are consistent with our earlier observations that the EBC matrix requires adjustment for there to be any validity to immunoassays.
-
Exhaled Breath Condensate an overview
Immunology and Allergy Clinics of North America, 2007Co-Authors: John F HuntAbstract:Exhaled Breath Condensate (EBC) is a promising source of biomarkers of lung disease. EBC is not a biomarker, but rather a matrix in which biomarkers may be identified, in that way equivalent to blood, sweat, tears, urine, and saliva. EBC may be thought of either as a body fluid or as a Condensate of Exhaled gas. The field of EBC research has advanced gradually, with the debates surrounding an emerging field helping to pose questions and gradually leading to answers. Conscientous assay technique will likely find in EBC any substance of substantially high enough concentration in the airway lining fluid.
-
identification of acid reflux cough using serial assays of Exhaled Breath Condensate ph
Cough, 2006Co-Authors: John F Hunt, James A Burns, Benjamin Gaston, Lina Ngamtrakulpanit, Dorothy Bunyan, Brian K Walsh, Alison Smith, Stephanie HomAbstract:Chronic cough is a common problem, frequently caused or exacerbated by acid reflux. Diagnosis of acid reflux cough is haphazard currently, often relying on long therapeutic trials of expensive medications. We tested the hypothesis that the most relevant mechanistic component of acid reflux in chronic cough is when it rises to the level of the airway where acid can potentially be aspirated. We further wished to determine if multi-sample Exhaled Breath Condensate (EBC) pH profiles can identify chronic cough patients likely to respond to proton pump inhibitor therapy. 59 subjects were recruited for this study. Initially we examined EBC pH (gas-standardized with Argon) in the setting of 15 experimental pharyngeal acid challenges to determine duration of EBC acidification. Subsequently, we enrolled 22 healthy subjects to determine a normal multi-sample Exhaled Breath Condensate pH profile over 1–3 days. We additionally obtained multi-sample EBC pH profiles in 22 patients with chronic cough. These samples were timed to occur after coughing episodes. Exhaled Breath Condensate pH was measured after gas standardization. We found that Exhaled Breath Condensate pH is substantially reduced for approximately 15 minutes after pharyngeal acid load. Healthy subjects rarely have any low EBC pH values (defined as < 7.4 based on a normative reference range from 404 healthy subjects). Patients with chronic cough who subsequently responded well to proton pump inhibition (n = 8) invariably had one or more cough episodes associated with EBC acidification. No patient who had normal EBC pH with each of their cough episodes reported a clinically relevant response to proton-pump inhibition. Patients whose cough responds to proton pump inhibition have transient Exhaled Breath Condensate acidification with coughing episodes, supporting the role of airway acidification in reflux-triggered cough. Multi-sample EBC pH profiles, involving samples collected immediately subsequent to a coughing episode, may be useful appropriately to direct therapy to those patients with cough who have relevant acid reflux.
-
normative data for ph of Exhaled Breath Condensate
Chest, 2006Co-Authors: Alix Pagetbrown, Lina Ngamtrakulpanit, Dorothy Bunyan, Alison Smith, Ashley Nguyen, John F HuntAbstract:Introduction Measurement of pH is one of the simplest and most technically validated biomarkers studied in Exhaled Breath Condensate (EBC). The pH of EBC has been found to be lower than controls in many respiratory disorders. Published data from normal control subjects have been reasonably consistent, but the data sets are not large. This study was undertaken to establish normative EBC pH reference values. Participants Four hundred four healthy subjects of all ages were enrolled. Interventions Each participant provided a single EBC sample using a disposable collector at modest temperature so that EBC was collected as a liquid. Measurements and results Samples of EBC were bubbled with argon gas to standardize for carbon dioxide, and pH was recorded with a calibrated and validated glass microelectrode on stabilization. The median EBC pH was 8.0 with interquartile (25 to 75%) range of 7.8 to 8.1. There were no differences based on age, sex, or race. The distribution is skewed, with 6.4% of EBC samples having a pH range Conclusions An extensive normal data set now exists that reveals EBC pH is maintained in a modestly alkaline and tight range in subjects who consider themselves healthy.
-
normative data for ph of Exhaled Breath Condensate
Chest, 2006Co-Authors: Alix Pagetbrown, Lina Ngamtrakulpanit, Dorothy Bunyan, Alison Smith, Stephanie Hom, Ashley Nguyen, John F HuntAbstract:Introduction Measurement of pH is one of the simplest and most technically validated biomarkers studied in Exhaled Breath Condensate (EBC). The pH of EBC has been found to be lower than controls in many respiratory disorders. Published data from normal control subjects have been reasonably consistent, but the data sets are not large. This study was undertaken to establish normative EBC pH reference values. Participants Four hundred four healthy subjects of all ages were enrolled. Interventions Each participant provided a single EBC sample using a disposable collector at modest temperature so that EBC was collected as a liquid. Measurements and results Samples of EBC were bubbled with argon gas to standardize for carbon dioxide, and pH was recorded with a calibrated and validated glass microelectrode on stabilization. The median EBC pH was 8.0 with interquartile (25 to 75%) range of 7.8 to 8.1. There were no differences based on age, sex, or race. The distribution is skewed, with 6.4% of EBC samples having a pH range Conclusions An extensive normal data set now exists that reveals EBC pH is maintained in a modestly alkaline and tight range in subjects who consider themselves healthy.
H Wirtz - One of the best experts on this subject based on the ideXlab platform.
-
multiplex analysis of cytokines in Exhaled Breath Condensate
Cytometry Part A, 2006Co-Authors: Ulrich Sack, Hartmut Kuhn, S Hammerschmidt, H Wirtz, Robert Scheibe, Michael Wotzel, Frank Emmrich, Gerard Hoheisel, C GessnerAbstract:Background: To improve monitoring of lung diseases, we analyzed cytokines in Exhaled Breath Condensate (EBC). The main challenge in measurement of cytokines in EBC is the low protein content, which requires concentration steps that conflict with the need for excessive fluid required by most commonly used kits. Methods: Here, a multiplex bead array for the detection of interleukins (IL) -1b, -6, -8, -10, TNF-a, and IL-12p70 was modified and validated for analysis in EBC samples. Furthermore, 33 healthy volunteers and 11 patients with acute lung injury were investigated. Results: In patients with inflammatory lung diseases, cytokine levels for all investigated cytokines were higher in comparison to healthy smokers or healthy volunteers. Discussion: Multiplexed immunoassays in highly sensitive approaches allow for cytokine detection in EBC. We found significant differences between patients and controls for all investigated cytokines. q 2006 International Society for Analytical Cytology
-
Exhaled Breath Condensate cytokine patterns in chronic obstructive pulmonary disease
Respiratory Medicine, 2005Co-Authors: C Gessner, Hartmut Kuhn, S Hammerschmidt, Ulrich Sack, Robert Scheibe, Michael Wotzel, Lothar Engelmann, G Hoheisel, Adrian Gillissen, H WirtzAbstract:Differences in cytokine patterns in stable chronic obstructive pulmonary disease (COPD), exacerbated COPD, smokers without apparent COPD, and healthy volunteers should be of interest for pathophysiological and therapeutic reasons. Methods including lavage, biopsy and sputum have been employed to investigate cytokines in the lung. For asystematic comparison, Exhaled Breath Condensate (EBC) appears to be well suited. We investigated healthy volunteers, smokers without apparent COPD, stable and exacerbated COPD patients (+/- inhalative steroids) and finally those whose exacerbation made mechanical ventilation inevitable, for a more complete picture of inflammatory cytokines in COPD. We chose EBC because it is non-invasive and can be used repeatedly in spontaneous Breathing individuals and during mechanical ventilation. EBC cytokines (IL-1 beta, IL-6, IL-8, IL-10, IL-12 p 70, TNF-alpha) were assayed from a single sample using a multiplex array test kit. We observed a significant increase of all cytokines in acute exacerbation compared to stable COPD, smokers, and volunteers. Stable COPD and volunteers exhibited only small differences in cytokine pattern with respect to IL-1 beta and IL-12 (P<0.01). Smokers had increased levels of all investigated cytokines (P<0.01) compared to non-smokers and, with the exception of IL-1 beta, to stable COPD. Inhaled steroids resulted in reduced levels of IL-1 beta, IL-6, IL-8, IL-10, and IL-12 (all: P<0.01) in stable COPD (all: ex-smokers) with dose dependency for IL-8, IL-1 beta and IL-12. EBC analysis successfully characterized important differences in stable COPD compared to exacerbation or smoking and non-smoking healthy individuals.
-
detection of p53 gene mutations in Exhaled Breath Condensate of non small cell lung cancer patients
Lung Cancer, 2004Co-Authors: C Gessner, Hartmut Kuhn, Katja Toepfer, S Hammerschmidt, Joachim Schauer, H WirtzAbstract:Abstract Early diagnosis of lung carcinoma is greatly desired. A potential source of early information regarding the process of cancerisation in the airways is Exhaled Breath Condensate (EBC). The direct approach to detecting cancerisation is examining DNA from the area of chronic damage, i.e. airways and lung parenchyma. We therefore investigated DNA in EBC of patients with NSCLC and healthy volunteers. Human DNA was amplified by PCR in Exhaled Breath Condensate and used to detect p53 mutations. A PCR of the β-actin gene fragment was used to detect human DNA in each of the EBC samples. In 65.7% of the samples, the β-actin gene was found. Extracted DNA as well as native EBC were equally suited as starting material for amplification. Mutations of the p53 gene were investigated in all EBC samples of NSCLC patients. p53 exons 5–8 were amplified using nested PCR and subsequently sequenced. Mutations were found in four of the patients ( n =11; 36.4%) while no mutation was found in volunteers ( n =10). Mutations detected in EBC were also compared with those of corresponding tumor tissue. Different point mutations in EBC and tumor tissue were revealed in all cases. Our findings demonstrate that Exhaled Breath Condensate may be used for analysis of somatic gene mutations in an area of direct tobacco-related DNA damage.
-
Exhaled Breath Condensate acidification in acute lung injury
Respiratory Medicine, 2003Co-Authors: C Gessner, Hartmut Kuhn, S Hammerschmidt, Joachim Schauer, Ulrich Sack, H J Seyfarth, Lothar Engelmann, H WirtzAbstract:Abstract Lung injury in ventilated lungs may occur due to local or systemic disease and is usually caused by or accompanied by inflammatory processes. Recently, acidification of Exhaled Breath Condensate pH (EBC-pH) has been suggested as marker of inflammation in airway disease. We investigated pH, ammonia, lactate, pCO 2 , HCO 3 − , IL-6 and IL-8 in EBC of 35 ventilated patients (AECC-classification: ARDS: 15, ALI: 12, no lung injury: 8). EBC-pH was decreased in ventilated patients compared to volunteers (5.85±0.32 vs. 7.46±0.48; P 4 + , lactate, HCO 3 − , pCO 2 , IL-6 and IL-8 were analyzed in EBC and correlated with EBC-pH. We observed correlations of EBC-pH with markers of local (EBC IL-6: r =−0.71, P r =−0.68, P r =−0.73, P 2 /FiO 2 ; r =0.54, P 4 + were found to correlate with EBC-pH. Inflammation-induced disturbances of regulatory mechanisms, such as glutaminase systems may result in EBC acidification. EBC-pH is suggested to represent a marker of acute lung injury caused by or accompanied by pulmonary inflammation.