~?\Failla, M. Biondi, G. Provvidenza Pistorio, M. Gili, E. Mastruzzo, C. Vancheri, C. Crimi, N.2006XIntranasal steroid reduces exhaled bronchial cysteinyl leukotrienes in allergic patients325-30Clin Exp Allergy363MarSummary Background Allergic rhinitis (AR) precedes and is often associated with bronchial asthma. Indeed, local and systemic inflammations in both conditions are very similar. Cysteinyl-leukotrienes (cys-LTs) are generated during early- and late-phase allergic reactions and induce smooth-muscle contraction, microvascular leakage, and mucous hypersecretion. Cys-LTs are detected in exhaled breath condensate (EBC) of asthmatics and regardless of bronchial symptoms, they are also found in EBC of rhinitic patients. Objective To evaluate cys-LTs in EBC of allergic patients and to assess the activity of nasal fluticasone propionate (FP) on EBC cys-LTs levels. Methods Cys-LTs coefficient of variation (CV) was evaluated from different EBC in 5 healthy volunteers. Cys-LTs levels from EBCs in 13 healthy controls and 56 allergic rhinitic (n=31) and rhinitic/asthmatic (n=25) patients were also evaluated at baseline. Subsequently patients were randomized to receive either FP 100 mug/day per nostril or placebo for 2 weeks and then re-evaluated for EBC cys-LTs. Results The CV was 14.12%. EBC cys-LTs in allergic patients were significantly higher than in healthy subjects (70.9 vs. 20.6 pg/mL (median), P<0.05), while it did not differ between asthmatic/rhinitic and purely rhinitic patients. Treatment significantly reduced cys-LTs (from 93.6 to 19.9 pg/mL, P<0.001). This effect was evident both in asthmatic/rhinitic and in rhinitic patients. Conclusion Treatment of AR with FP significantly reduces the levels of cys-LTs, major noninvasive markers of lower airway inflammation, suggesting that upper and lower airway inflammation is present and should be thus treated as a whole in subjects with AR with and without asthma.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16499643 !0954-7894 (Print) Journal Article16499643yDepartment of Internal and Specialistic Medicine, Section of Respiratory Medicine, University of Catania, Catania, Italy.~?hRobroeks, C. M. Jobsis, Q. Damoiseaux, J. G. Heijmans, P. H. Rosias, P. P. Hendriks, H. J. Dompeling, E.2006RCytokines in exhaled breath condensate of children with asthma and cystic fibrosis349-55Ann Allergy Asthma Immunol962FebzBACKGROUND: Inflammatory mediators in exhaled breath condensate (EBC) indicate ongoing inflammation in the lungs and might differentiate between asthma and cystic fibrosis (CF). OBJECTIVES: To evaluate the presence, concentration, and short-term variability of TH1- and TH2-mediated cytokines (interferon-gamma [IFN-gamma], tumor necrosis factor alpha [TNF-alpha], interleukin 10 [IL-10], IL-5, IL-4, and IL-2) in EBC of children with asthma or CF and in controls and to analyze the discriminating ability of inflammatory markers in EBC between children with asthma or CF and controls. METHODS: Expired air was conducted through a double-jacketed glass tube cooled by circulating ice water. In 33 asthmatic children, 12 children with CF, and 35 control children, EBC was collected during tidal breathing. Cytokines were measured using flow cytometry. RESULTS: Interleukin 2, IL-4, IFN-gamma, and IL-10 were detected in 16%, 16%, 11%, and 9%, respectively, of all samples in asthma and CF. Interleukin 5 and TNF-alpha were not detected in children with CF. Cytokine concentrations did not differ significantly in children with asthma vs CF. In controls, IFN-gamma, TNF-alpha, and IL-10 were detected in 9%, 14%, and 3%, respectively; IL-2, IL-4, and IL-5 were not detected in controls. CONCLUSIONS: Cytokines such as IFN-gamma, TNF-alpha, IL-10, IL-5, IL-4, and IL-2 can be detected in EBC of children with asthma or CF. However, the concentrations found are close to the detection limits of the assay used. These findings emphasize the importance of developing more sensitive techniques for the analysis of EBC and of standardizing the EBC collection method.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16498859 !1081-1206 (Print) Journal Article16498859sDepartment of Paediatric Pulmonology, University Hospital Maastricht, Maastricht, The Netherlands. crob@paed.azm.nl~?iSack, U. Scheibe, R. Wotzel, M. Hammerschmidt, S. Kuhn, H. Emmrich, F. Hoheisel, G. Wirtz, H. Gessner, C.2006<Multiplex analysis of cytokines in exhaled breath condensate169-172 Cytometry A69A3Feb 22BACKGROUND: 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) -1beta, -6, -8, -10, TNF-alpha, 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. (c) 2005 International Society for Analytical Cytology.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16496377 !1552-4922 (Print) Journal article16496377cInstitute of Clinical Immunology and Transfusion Medicine, University of Leipzig, Leipzig, Germany. ~?Griesenbach, U. Kitson, C. Escudero Garcia, S. Farley, R. Singh, C. Somerton, L. Painter, H. Smith, R. L. Gill, D. R. Hyde, S. C. Chow, Y. H. Hu, J. Gray, M. A. Edbrooke, M. Ogilvie, V. Macgregor, G. Scheule, R. K. Cheng, S. H. Caplen, N. J. Alton, E. W.2006gCationic lipid-mediated siRNA and antisense oligonucleotide transfer to airway epithelial cells in vivo26 Respir Res71Feb 15KABSTRACT: BACKGROUND: The cationic lipid Genzyme lipid (GL) 67 is the current gold-standard for in vivo lung gene transfer. Here, we assessed, if GL67 mediated uptake of siRNAs and asODNs into airway epithelium in vivo. Methods: Anti-lacZ and ENaC siRNA and asODN were complexed to GL67 and administered to the mouse airway epithelium in vivo Transfection efficiency and efficacy were assessed using real-time RT-PCR as well as through protein expression and functional studies. Results: In vitro, GL67 efficiently complexed asODNs and siRNAs, and both were stable in exhaled breath condensate. Importantly, during in vitro selection of functional siRNA and asODN we noted that asODNs accumulated rapidly in the nuclei of transfected cells, whereas siRNAs remained in the cytoplasm, a pattern consistent with their presumed site of action. Following in vivo lung transfection siRNAs were only visible in alveolar macrophages, whereas asODN also transfected alveolar epithelial cells, but no significant uptake into conducting airway epithelial cells was seen. SiRNAs and asODNs targeted to SZ-galactosidase reduced gal mRNA levels in the airway epithelium of K18-lacZ mice by 30% and 60%, respectively. However, this was insufficient to reduce protein expression. In an attempt to increase transfection efficiency of the airway epithelium, we increased contact time of siRNA and asODN using the in vivo mouse nose model. Although highly variable and inefficient, transfection of airway epithelium with asODN, but not siRNA, was now seen. As asODNs more effectively transfected nasal airway epithelial cells, we assessed the effect of asODN against the epithelial sodium channel (ENaC), a potential therapeutic target in cystic fibrosis; no decrease in ENaC mRNA levels or function was detected. Conclusions: This study suggests that although siRNAs and asODNs can be developed to inhibit gene expression in culture systems and certain organs in vivo, barriers to nucleic acid transfer in airway epithelial cells seen with large DNA molecules may also affect the efficiency of in vivo uptake of small nucleic acid molecules.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16480492 &1465-993X (Electronic) Journal article16480492~?ZPaget-Brown, A. O. Ngamtrakulpanit, L. Smith, A. Bunyan, D. Hom, S. Nguyen, A. Hunt, J. F.20062Normative data for pH of exhaled breath condensate426-30Chest1292FebINTRODUCTION: 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 < 7.4. 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.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16478862 !0012-3692 (Print) Journal Article16478862jDivision of Neonatology, Department of Pediatrics, University of Virginia, Charlottesville, VA 22908, USA.~?xCarpagnano, G. E. Resta, O. Ventura, M. T. Amoruso, A. C. D. I. Gioia G Giliberti, T. Refolo, L. Foschino-Barbaro, M. P.2006kAirway inflammation in subjects with gastro-oesophageal reflux and gastro-oesophageal reflux-related asthma323-31 J Intern Med2593MarStudy objectives. Asthma and gastro-oesophageal reflux (GER) are both characterized by airway inflammation. Design. The purposes of this work were (i) to study airway inflammation in patients troubled by gastro-oesophageal reflux (GER) and GER associated with asthma, (ii) to ascertain whether GER can aggravate asthma by exacerbating the pre-existing airway inflammation and oxidative stress and (iii) to establish the validity of analysing breath condensate and induced sputum when studying the airways of subjects affected by GER. Patient s and methods. We enrolled 14 patients affected by mild asthma associated with GER (40 +/-12 years), nine with mild but persistent asthma (39 +/- 13 years), eight with GER (35 +/- 11 years) and 17 healthy subjects (37 +/- 9 years). Sputum cell counts and concentrations of interleukin-4 (IL-4), IL-6 and 8-isoprostane were measured in breath condensate and supernatant. Measurements and results. GER-related asthma is characterized by an eosinophilic inflammation, as determined by elevated concentrations of IL-4 in breath condensate and sputum supernatant, and by sputum cell analysis. GER alone presents a neutrophilic pattern of inflammation when determined by elevated concentrations of IL-6 in sputum cell analysis. A concomitant increase has been found in 8-isoprostane in GER associated (or not associated) with asthma. Conclusions. We conclude that GER is characterized by a neutrophilic airway inflammation and by increased oxidative stress. GER does not however aggravate pre-existing airway inflammation in asthma patients. Determinations of inflammatory and oxidant markers in the breath condensate of subjects with GER reflect these measured in the induced sputum.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16476110 !0954-6820 (Print) Journal Article16476110GInstitute of Respiratory Diseases, University of Foggia, Foggia, Italy.A~?Hunt, J.2006RExhaled Breath Condensate pH: Reflecting Acidification of the Airway at All Levels366-7Am J Respir Crit Care Med1734Feb 15fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16467175 !1073-449X (Print) Journal Article16467175 F~?9Davidsson, A. Soderstrom, M. Sjosward, K. N. Schmekel, B.2006LChlorine in Breath Condensate - A Measure of Airway Affection in Pollinosis? RespirationFeb 1Background: Infiltration of inflammatory cells in bronchial mucosa and glandular hypersecretion are hallmarks of asthma. It has been postulated that exhaled breath condensate (EBC) mirrors events in epithelial lining fluid of airways, such as presence of local inflammation as well as glandular hypersecretion. It is also well known that eosinophil cationic protein (ECP) and cysteinyl-leukotrienes (cys-LT) are released by circulating inflammatory cells when triggered by antigen stimulation in asthma patients. Objectives: The aim of this study was to evaluate whether chlorine and/or cys-LT in EBC would reflect changes of exposure of airborne pollen in patients with asthma. Methods: EBC and serum were collected from 23 patients with allergic asthma during a pollen season and repeated 5 months later during a period with no aeroallergens. Chlorine was measured by means of a sensitive coulometric technique and cys-LT by an EIA technique. Serum ECP was measured and lung function tests were performed and symptoms noted during both occasions. Results: Significantly higher concentrations of chlorine in EBC (p = 0.007) and ECP in serum (p = 0.003) were found during the pollen season compared to post-season. Chlorine levels tended to be higher in patients who reported of chest symptoms compared to those who denied symptoms during the pollen season (p = 0.06). Areas under the receiver-operated characteristic curves (AUC(ROC)) were compared and similar discriminative power to identify exacerbations of asthma was recorded by chlorine in EBC (range 0.67-0.78) and ECP in serum (range 0.64-0.78). Conclusion: It is concluded that chlorine in EBC and ECP in serum decreased significantly post-season, and this is suggested to mirror the decrement in airborne antigen. It is furthermore proposed that chlorine in EBC and ECP in serum tend to have a similar capacity to identify seasonal variations in airborne pollen in patients with asthma. Copyright (c) 2006 S. Karger AG, Basel.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16462136 !0025-7931 (Print) Journal article16462136Faculty of Health Sciences, Department of Medicine and Care, Section of Clinical Physiology, University Hospital, Linkoping, Sweden.gF~? $Spicuzza, L. Di Maria, G. Polosa, R.20067Adenosine in the airways: Implications and applicationsEur J PharmacolFeb 2Adenosine in a signaling nucleoside eliciting many physiological responses. Elevated levels of adenosine have been found in bronchoalveolar lavage, blood and exhaled breath condensate of patients with asthma a condition characterized by chronic airway inflammation. In addition, inhaled adenosine-5'-monophosphate induces bronchoconstriction in asthmatics but not in normal subjects. Studies on animals and humans have shown that bronchoconstriction is most likely due to the release of inflammatory mediators from mast cells. However a number of evidences suggest that adenosine modulates the function of many other cells involved in airway inflammation such as neutrophils, eosinophils, lymphocytes and macrophages. Although this clear pro-inflammatory role in the airways, adenosine may activate also protective mechanisms particularly against lung injury. For many years this dual role of adenosine in the respiratory system has represented an enigma, and only recently it has become clear that biological functions of adenosine are mediated by four distinct subtypes of receptors (A(1), A(2A), A(2B), and A(3)) and that biological responses are determined by the different pattern of receptors distribution in specific cells. Therefore, pharmacological modulation of adenosine receptors, particularly A(2B), may represent a novel therapeutic approach for inflammatory diseases. Moreover, as bronchial response to adenosine strictly reflects airway inflammation in asthma, bronchial challenge with adenosine is considered a valuable clinical tool to monitor airway inflammation, to follow the response to anti-inflammatory treatments and to help in the diagnostic discrimination between asthma and chronic obstructive lung disease.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16458886 !0014-2999 (Print) Journal article16458886uDipartimento di Medicina Interna e Medicina Specialistica Sezione Malattie Respiratorie-Universita di Catania, Italy.V~? Pobed'onna, H. P.2005|[Antioxidant protection, metabolites of nitrogen oxide on the forming of oxidative stress in patients with bronchial asthma]36-40 Lik Sprava5-6kAdult Antioxidants/*metabolism Asthma/blood/*metabolism/physiopathology Biological Markers/analysis Breath Tests Bronchi/physiopathology Comparative Study English Abstract Female Forced Expiratory Volume/physiology Humans Lipid Peroxidation/physiology Male Nitrates/*analysis Nitric Oxide/*metabolism Nitrites/*analysis *Oxidative Stress Severity of Illness IndexJul-Sep276 Patients with bronchial asthma of different degree of severity have been observed. An increase in processes of lipid peroxidation, depression of antioxidant protection, increase in oxide nitrogen metabolites in blood serum and condensate of exhaled air were detected. These pathological changes may be considered as manifestation of system oxidative stress more expressed in bronchi.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16396289 !1019-5297 (Print) Journal Article16396289~? NKo, F. W. Lau, C. Y. Leung, T. F. Wong, G. W. Lam, C. W. Lai, C. K. Hui, D. S.2006lExhaled breath condensate levels of eotaxin and macrophage-derived chemokine in stable adult asthma patients44-51Clin Exp Allergy361JanBACKGROUND: Asthma is associated with esoinophilic airway inflammation and overproduction of T-helper type 2 (Th2) lymphocyte-related cytokines. OBJECTIVE: This study assessed the eosinophil chemoattractant eotaxin and Th2-specific macrophage-derived chemokine (MDC) in the adult asthmatic airway. Eotaxin and MDC levels were determined in exhaled breath condensate (EBC) obtained from adult patients with asthma. METHODS: Fifty-four asthmatics (20 male, mean (SD) age 40 (12) years and percentage predicted forced expiratory volume in 1 s (FEV(1)) 81.7 (20.8)) and 20 age- and sex-matched controls were studied. EBC was collected using EcoScreen by 10 min of tidal breathing with a nose clip. Concentrations of eotaxin and MDC were measured by ELISA. RESULTS: Asthma patients on inhaled corticosteroid (ICS) had a higher median interquartile range (IQR) level of eotaxin than the steroid-naive asthmatics (18.5 (17.7-20.1) vs. 17.9 (17.0-18.6) pg/mL, P=0.02) and controls (18.5 (17.7-20.1) pg/mL vs 17.4 (16.3-18.0) pg/mL, P=0.001). Eotaxin level in EBC had a significant negative correlation with the FEV(1)/forced vital capacity ratio (r=-0.43, P=0.03) in steroid-naive asthmatics. EBC MDC level was higher in subjects on ICS than the steroid naive asthmatics (120 (118-125) vs. 117 (116-119) pg/mL, P=0.01) and the controls (120 (118-125) vs. 117 (116-120) pg/mL, P=0.02). CONCLUSIONS: Eotaxin and MDC could be measured in EBC of adults with asthma. EBC eotaxin and MDC levels were higher in asthmatics on ICS than the steroid-naive asthmatics or controls. Exhaled chemokines may be potential non-invasive markers for assessing airway inflammation in asthmatics.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16393265 !0954-7894 (Print) Journal Article16393265Departments of Medicine and Therapeutics, Paediatrics and Chemical Pathology, The Chinese University of Hong Kong, Hong Kong, China. fannyko@cuhk.edu.hk~? QMoeller, A. Franklin, P. Hall, G. L. Horak, F., Jr. Wildhaber, J. H. Stick, S. M.2006/Measuring exhaled breath condensates in infants184-7Pediatr Pulmonol412Feb*There is growing interest in investigating compounds of exhaled breath condensates (EBC) as potential noninvasive markers of airways disease processes. Some of these markers have the potential to provide information on the early stages of disease. In this paper, we present a method for collecting EBC during both oral and nasal breathing in infants. Fifty-four infants (mean age, 13.3 months; range, 1-30 months) undergoing infant lung-function testing were recruited for this study. Breath condensates were collected during sedated sleep, using a custom-made collection device. Collections were made for 10 min during normal tidal breathing. Nasal measurements were attempted in all children by placing a face-mask over the nose and mouth and keeping the mouth closed. In 14 infants, oral measurements were made by placing a face-mask over the mouth only and occluding the nose. Condensates were collected successfully in all but one child. The collected volume ranged from 50-550 microl (mean +/- SD, 281.8 +/- 145.8 microl). The volume of EBC collected was correlated to age, length, weight, and minute ventilation. Significantly more EBC was collected during oral compared to nasal breathing (354.3 vs. 277.5 microl, P = 0.03). There were no significant changes in heart rate, respiratory rate, or oxygen saturation during collection. The collection of EBC in young children and infants is feasible and safe, and the method used here allows the successful collection of reasonable amounts of exhaled condensate. Pediatr Pulmonol. 2006; 41:184-187. (c) 2005 Wiley-Liss, Inc.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16372354 !8755-6863 (Print) Journal Article16372354zDepartment of Respiratory Medicine, Princess Margaret Hospital for Children, Subiaco, Perth, Western Australia, Australia.~? vBaraldi, E. Giordano, G. Pasquale, M. F. Carraro, S. Mardegan, A. Bonetto, G. Bastardo, C. Zacchello, F. Zanconato, S.2006q3-Nitrotyrosine, a marker of nitrosative stress, is increased in breath condensate of allergic asthmatic children90-6Allergy611JanBACKGROUND: Asthmatic patients have high exhaled nitric oxide (NO) levels. NO-mediated inflammatory actions are mainly due to NO conversion into reactive nitrogen species, which can lead to nitrotyrosine formation. The aim of this study was to assess 3-nitrotyrosine (3-NT) levels in exhaled breath condensate (EBC) of asthmatic and healthy children and to investigate whether there is any relationship with exhaled NO (FE(NO)) and lung function. METHODS: The study included 20 asthmatic children (10 steroid-naive with intermittent asthma, 10 steroid-treated with unstable persistent asthma) and 18 healthy controls. They underwent FE(NO) measurement, EBC collection and spirometry. 3-NT was measured by a new liquid chromatography-tandem mass spectrometry (LC-MS/MS) method in isotopic dilution. RESULTS: The median EBC concentration of 3-NT (expressed as nitrotyrosine/tyrosine ratio x 100) in asthmatic children was fivefold higher than in healthy subjects [0.23% (0.12-0.32) vs 0.04% (0.02-0.06), P < 0.001] with no difference between steroid-naive and unstable steroid-treated asthmatic patients. FE(NO) levels were higher in asthmatic [44.6 ppb (36.0-66.0)] than in healthy children [7.5 ppb (6.0-8.8), P < 0.001]. No correlation was found among 3-NT, FE(NO) and lung function parameters. CONCLUSION: Nitrotyrosine is high in EBC of asthmatic children and could be considered as a noninvasive marker of nitrosative events in the airways.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16364162 !0105-4538 (Print) Journal Article16364162>Department of Pediatrics, University of Padova, Padova, Italy. ~?P, M. I, C. Je, R. N, N. Ab, R.2005[[pH breath condensate measurement on asthmatic patients - Comparing two different methods.]20-1Rev Port Pneumol11 6 Suppl 1Nov-Dec Background: Recent publications have shown that pH breath condensate (BC) is a good bronchial inflammation marker and could be used on the study of respiratory diseases, namely asthma. There are some mean pH BC published and according with some authors the mean pH value for controls is 7.65 +/-0.2, 5.23 +/-0.2 in asthma attacks and 7.8 +/-0.1 for stable asthmatics1. At the moment there are two devices to collect BC: the R Tube(R) and the Ecoscreen(R). Objective: To compare the pH BC results, collected from asthmatic patients with R Tube(R) and Ecoscreen(R). Methods: 15 medicated asthmatic patients, sensitized to at least one airborne allergen were selected from the Immunoallergy Department - Dona Estefania Hospital. All the patients were submitted to body plethysmography, exhaled nitric oxide (eNO) and BC collection with R Tube(R) and EcoScreen(R) at the Pathophysiology Department, Faculdade de Ciencias Medicas, UNL. pH was measured on the BC. Results: 7 patients were male and 8 female. The median age was 14.27 years +/-4.12 (9-22 years). The median pH R Tube(R) BC was 8.07 +/-1.23, at a median temperature of 26.66 masculineC +/-1.49. For Ecoscreen(R), the median pH BC was 7.78 +/-1.42, at a median temperature of 26.69 masculineC +/-1.55. Comparing the pH results, there are no statistically differences between both methods. The mean of the differences between the two methods was 0.28. We didnt find any relation between pH BC values and eNO measurements or body plethysmography results. Conclusions: Although the small number of patients studied, the pH BC results were similar for both methods and similar of those published for stable asthmatics. 1. Hunt JF et al. Endogenous airway acidification. Am J Resp Crit Care Med 2000; 161: 694-699. Granted by: Fundacao Calouste Gulbenkian, SaudAr Project.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16362100 !0873-2159 (Print) Journal Article16362100XMedicao do pH no condensado bronquico de doentes asmaticos - Comparacao de dois metodos.Servico de Imunoalergologia do Hospital de Dona Estefania Departamento Universitario de Fisiopatologia, Faculdade de Ciencias Medicas - UNL Centro de Estudos de Patologia Respiratoria - FCT.x~?Mansoor, J. K. Morrissey, B. M. Walby, W. F. Yoneda, K. Y. Juarez, M. Kajekar, R. Severinghaus, J. W. Eldridge, M. W. Schelegle, E. S.2005_L-arginine supplementation enhances exhaled NO, breath condensate VEGF, and headache at 4,342 m289-300High Alt Med Biol64WinterWe examined the effect of dietary supplementation with L-arginine on breath condensate VEGF, exhaled nitric oxide (NO), plasma erythropoietin, symptoms of acute mountain sickness, and respiratory related sensations at 4,342 m through the course of 24 h in seven healthy male subjects. Serum L-arginine levels increased in treated subjects at time 0, 8, and 24 h compared with placebo, indicating the effectiveness of our treatment. L-arginine had no significant effect on overall Lake Louise scores compared with placebo. However, there was a significant increase in headache within the L-arginine treatment group at 12 h compared with time 0, a change not seen in the placebo condition between these two time points. There was a trend (p = 0.087) toward greater exhaled NO and significant increases in breath condensate VEGF with L-arginine treatment, but no L-arginine effect on serum EPO. These results suggest that L-arginine supplementation increases HIF-1 stabilization in the lung, possibly through a NO-dependent pathway. In total, our observations indicate that L-arginine supplementation is not beneficial in the prophylactic treatment of AMS.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16351563 !1527-0297 (Print) Journal Article16351563-University of the Pacific, Stockton, CA, USA.g~?iCarpagnano, G. E. Foschino Barbaro, M. P. Cagnazzo, M. Di Gioia, G. Giliberti, T. Di Matteo, C. Resta, O.2005oUse of exhaled breath condensate in the study of airway inflammation after hypertonic saline solution challenge3159-66Chest12856Aged Asthma/*diagnosis *Breath Tests Cell Count Comparative Study Female Humans Hydrogen-Ion Concentration Inflammation Mediators/*analysis Interleukin-6/*analysis Male Middle Aged Pulmonary Disease, Chronic Obstructive/*diagnosis Saline Solution, Hypertonic/diagnostic use Tumor Necrosis Factor-alpha/analysisNov)STUDY OBJECTIVES: Hypertonic saline solution inhalation is suspected to produce airway inflammation. DESIGN: The aim of this study was to verify this hypothesis by measuring inflammatory markers in exhaled breath condensate (EBC) collected before and after sputum induction with hypertonic and isotonic saline solution. PATIENTS AND METHODS: We enrolled 10 patients with asthma, 10 patients with COPD, and 7 healthy subjects with no history of lung disease. Levels of interleukin (IL)-6 and tumor necrosis factor (TNF)-alpha were measured in EBC by a specific enzyme immunoassay kit. Exhaled pH was measured after deaeration/decarbonation by bubbling with argon (350 mL/min) for 10 min by means of a pH meter. MEASUREMENTS AND RESULTS: Exhaled IL-6 and TNF-alpha concentrations were greater and pH was decreased compared to baseline after hypertonic saline solution inhalation in each group of subjects studied. No changes were observed following isotonic saline solution inhalation. Concentrations of IL-6, TNF-alpha, and pH in EBC correlated. CONCLUSIONS: These findings suggest that hypertonic saline solution inhalation could cause a low-grade inflammation in airways, and levels of inflammatory markers such as IL-6, TNF-alpha, and pH in EBC may be a useful noninvasive way to assess and monitor airway inflammation.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16304257 !0012-3692 (Print) Journal Article16304257UInstitute of Respiratory Disease, University of Foggia, Italy. ge.carpagnano@unifg.it~?ALeung, T. F. Li, C. Y. Yung, E. Liu, E. K. Lam, C. W. Wong, G. W.2006]Clinical and technical factors affecting pH and other biomarkers in exhaled breath condensate87-94Pediatr Pulmonol411JanExhaled breath condensate (EBC) pH appears to be a robust measure of asthma. However, the association between EBC pH and clinical factors and airway inflammatory markers remains unclear. The objectives of this study were to investigate the factors determining EBC pH in asthmatic children, and the reproducibility and effects of collection devices on EBC pH in nine healthy, nonsmoking adults. EBC was collected once from asthmatic children using EcoScreen, and from adults over 3 consecutive days using both RTubes and EcoScreen. EBC pH was measured immediately in non-deaerated samples by microelectrode pH meter. Concentrations of 8-isoprostane, cysteinyl leukotrienes (cys-LT), and leukotriene B4 (LTB4) were measured using enzyme immunoassay. Exhaled nitric oxide concentration (FeNO) was measured by chemiluminescence. Fifty-eight asthmatics (16 intermittent, 12 mild persistent, and 30 moderate-to-severe persistent) were recruited. EBC pH was lower among patients with moderate-to-severe persistent than intermittent asthma (P = 0.046). This marker correlated inversely with disease severity score (rho = -0.276, P = 0.036), but not FeNO or other EBC biomarkers. Bland-Altman analyses found pH but not other EBC biomarkers to be reproducible, which were confirmed by its low coefficient of variation (2.7%; range, 0.4-5.2%). There was poor correlation between pH in EBC collected by RTube and EcoScreen (rho = 0.059, P = 0.784). Factor analysis selected four factors that explained 67.5% of the total variance, and EBC pH clustered with both cys-LT and LTB4. In conclusion, our results suggest that pH in non-deaerated EBC is influenced by asthma severity in children. EBC pH measurement is reproducible, but is dependent on the collection devices used.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16292777 !8755-6863 (Print) Journal Article16292777fDepartment of Pediatrics, Chinese University of Hong Kong, Hong Kong SAR, China. leung2142@cuhk.edu.hkr~?NEffros, R. M. Casaburi, R. Su, J. Dunning, M. Torday, J. Biller, J. Shaker, R.2006PThe Effects of Volatile Salivary Acids and Bases on Exhaled Breath Condensate pH386-92Am J Respir Crit Care Med1734Feb 15uRationale: Recent studies have reported acidification of exhaled breath condensate (EBC) in inflammatory lung diseases. This phenomenon, designated "acidopnea," has been attributed to airway inflammation. Objectives: To determine whether salivary acids and bases can influence EBC pH in chronic obstructive pulmonary disease (COPD). Methods: Measurements were made of pH, electrolytes, and volatile bases and acids in saliva and EBC equilibrated with air in 10 healthy subjects and 10 patients. Results: The average EBC pH in COPD was reduced (normal, 7.24 +/- 0.24 SEM; range, 6.11-8.34; COPD, 6.67 +/- 0.18; range, 5.74-7.64; p = 0.079). EBCs were well buffered by NH(4)(+)/NH(3) and CO(2)/HCO(3)(-) in all but four patients, who had NH(4)(+) concentrations under 60 mumol/L, and acetate concentrations that approached or exceeded those of NH(4)(+). Saliva contained high concentrations of acetate ( approximately 6,000 mumol/L) and NH(4)(+) ( approximately 12,000 mumol/L). EBC acetate increased and EBC NH(4)(+) decreased when salivary pH was low, consistent with a salivary source for these volatile constituents. Nonvolatile acids did not play a significant role in determining pH of condensates because of extreme dilution of respiratory droplets by water vapor ( approximately 1:12,000). Transfer of both acetic acid and NH(3) from the saliva to the EBC was in the gas phase rather than droplets. Conclusions: EBC acidification in COPD can be affected by the balance of volatile salivary acids and bases, suggesting that EBC pH may not be a reliable marker of airway acidification. Salivary acidification may play an important role in acidopnea.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16284109 !1073-449X (Print) Journal Article16284109qLABiomed-Harbor-UCLA Medical Center, 1124 West Carson Street, RB2, Torrance, CA 90502-2064. reffros@labiomed.org.E~?.Torrego, A. Cimbollek, S. Hew, M. Chung, K. F.2005mNo effect of omeprazole on pH of exhaled breath condensate in cough associated with gastro-oesophageal reflux10Cough1Oct 19DBACKGROUND: Endogenous airway acidification evaluated as pH in exhaled breath condensate (EBC) has been described in patients with chronic cough. Proton pump inhibitors improve gastro-oesophageal reflux (GOR)-associated cough. METHODS: We examined pH levels in EBC and capsaicin cough response in 13 patients with chronic cough (mean age 41 years, SD 9) associated with GOR before and after omeprazole treatment (40 mg/day for 14 days) and its relationship with clinical response. RESULTS: Omeprazole abolished symptoms associated with GOR. Patients with chronic cough had an EBC pH of 8.28 (SD 0.13) prior to treatment but this did not change with omeprazole treatment. There was a significant improvement in the Leicester Cough Questionnaire symptom scores from 80.8 points (SD 13.2) to 95.1 (SD 17) (p = 0.02) and in a 6-point scale of cough scores, but there was no change in capsaicin cough response. CONCLUSION: An improvement in GOR-associated cough was not associated with changes in EBC pH or capsaicin cough response. These parameters are not useful markers of therapeutic response.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16270907 &1745-9974 (Electronic) Journal Article16270907Department of Thoracic Medicine, National Heart & Lung Institute, Imperial College, Royal Brompton Hospital, London, UK. a.torrego@imperial.ac.uk~?/de Lema, J. B. Gonzalez, M. Vigil, L. Casan, P.2005W[Exhaled breath condensate: standardized collection of samples from healthy volunteers]584-6Arch Bronconeumol4110gAdult Breath Tests/*methods English Abstract Exhalation Female Humans Male Specimen Handling/*standardsOctExpired breath condensate collection is a noninvasive technique for obtaining a sample in which to analyze substances that reflect the functional status of the lung and other tissues. Twenty healthy volunteers provided 3 expired breath samples: the second was collected 20 minutes after the first and the third 48 hours after the first. The air and condensate volumes were assessed. The mean (SD) volume of condensate in exhaled air over a period of 15 minutes was 1.8 (0.5) mL (95% confidence interval [CI], 1.5-2 mL) and the coefficient of variation was 29%. Analysis of variance in the 3 samples demonstrated no significant differences. The mean volume of air inhaled over 15 minutes was 119 (25) L (95% CI, 112-125 L). These results indicate that it takes at least 15 minutes and the inhalation of some 120 L of air to collect a condensate volume that exceeds 1.5 mL, sufficient to allow distribution in aliquots to analyze fundamental physical and chemical properties (conductivity, pH) and certain relevant biomarkers.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16266673 40300-2896 (Print) Journal Article Validation Studies16266673]Condensado de aire espirado: estandarizacion de la recogida de muestras en voluntarios sanos.Departament de Pneumologia, Hospital de la Santa Creu i de Sant Pau, Facultat de Medicina, Universitat Autonoma de Barcelona, Barcelona, Spain. jbruno@hsp.santpau.es t~?-Bruhn, A. Liberona, L. Lisboa, C. Borzone, G.2005[Limitations of the technique to determine hydrogen peroxide levels in exhaled breath condensate from patients with adult respiratory distress syndrome]542-6Arch Bronconeumol4110Breath Tests/methods English Abstract Exhalation Female Humans Hydrogen Peroxide/*analysis Male Middle Aged Respiratory Distress Syndrome, Adult/*metabolismOctOBJECTIVE: Exhaled breath condensate represents an alternative to bronchoalveolar lavage for the analysis of markers of inflammation and oxidative stress in patients with adult respiratory distress syndrome (ARDS). However, analysis of hydrogen peroxide (H2O2) yields variable results that do not correlate with severity of the clinical presentation. In an attempt to explain this variability, the aim of the present study was to assess the possible limitations of the most commonly used technique for analyzing H2O2 in breath condensate. PATIENTS AND METHODS: H2O2 levels were analyzed using the Gallati technique (linear range between 0.3 and 10 microM, r=0.99; P<.05) in serial samples of condensate taken from the expiratory tube of a mechanical ventilator in 6 patients with ARDS. RESULTS: The volume of condensate obtained correlated to minute ventilation (r=0.96; P<.05). In 11 out of 23 samples, a spectrophotometer reading was obtained at 450 nm despite the absence of the characteristic color of the reaction and in some of these samples a spontaneous reading was obtained that was indicative of contamination. The absorbance spectrum of these samples did not contain the characteristic peak for H2O2 at 450 nm and pretreatment of some samples with catalase did not affect the absorbance at 450 nm. CONCLUSIONS: The spectrophotometric method commonly used to measure H2O2 levels in breath condensate lacks specificity in ARDS due to the presence of variable levels of contaminants in the samples, which lead to false positives.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16266666 !0300-2896 (Print) Journal Article16266666Limitaciones de la tecnica de determinacion de peroxido de hidrogeno en el condensado del aire espirado de pacientes con sindrome de distres respiratorio del adulto.Departamento de Enfermedades Respiratorias, Facultad de Medicina, Pontificia Universidad Catolica de Chile, Santiago de Chile, Chile.~?Gonzalez-Mangado, N.2005C[Analysis of exhaled breath condensate: a technique with a future?]540-1Arch Bronconeumol41103Breath Tests/*methods Exhalation Forecasting HumansOctfhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16266665 *0300-2896 (Print) Comment Editorial Review162666659Analisis del condensado exhalado: una tecnica con futuro? ~?ABiernacki, W. A. Kharitonov, S. A. Biernacka, H. M. Barnes, P. J.2005WEffect of montelukast on exhaled leukotrienes and quality of life in asthmatic patients1958-63Chest1284EAcetates/*therapeutic use Anti-Asthmatic Agents/*therapeutic use Asthma/*drug therapy/physiopathology/psychology Breath Tests *Exhalation Family Practice Female Forced Expiratory Volume Humans Leukotrienes/*analysis Male Middle Aged *Quality of Life Quinolines/*therapeutic use Research Support, Non-U.S. Gov't Vital CapacityOctUSTUDY OBJECTIVES: In some patients with asthma treated with inhaled corticosteroids, suppression of inflammation is incomplete. This may be because the effect of corticosteroids on cysteinyl-leukotriene (cys-LT) biosynthesis is limited. Montelukast is a cys-LT antagonist that significantly improves asthma control in corticosteroid-treated asthmatic patients. However, not all patients treated with cys-LT antagonists show a clinical improvement. DESIGN: We have studied the effect of treatment for 4 weeks with montelukast (10 mg/d) on exhaled cys-LTs and leukotriene B4 (LTB4), exhaled nitric oxide, asthma quality of life (AQL), and respiratory function in patients with stable asthma. SETTING: Asthma clinics in general practice. PATIENTS: We studied 50 patients (30 men; mean +/- SEM age, 53 +/- 2 years) who were treated with inhaled corticosteroids. MEASUREMENTS AND RESULTS: We detected cys-LTs in exhaled breath condensate in 25 of 50 patients; however, in the normal nonasthmatic subjects, cys-LTs were below the limit of detection. After treatment with montelukast, there was a fall in cys-LT concentrations from 14.6 +/- 3.3 to 8.5 +/- 2.6 pg/mL after 2 weeks (p > 0.05) and to 3.9 +/- 1.3 pg/mL after 4 weeks (p < 0.01). Exhaled LTB4 levels were also elevated. After treatment with montelukast, LTB4 levels fell from 33.0 +/- 3.9 to 20.4 +/- 2.5 pg/mL after 2 weeks of treatment (p < 0.05), and to 17.0 +/- 2.2 pg/mL after 4 weeks of treatment (p < 0.01). These changes in exhaled cys-LT and LTB4 were associated with significant improvements in AQL scores. CONCLUSIONS: It appears that in some patients with stable asthma treated with inhaled corticosteroids, the suppression of inflammation is incomplete. Adding a leukotriene receptor antagonist can provide a complementary effect of controlling inflammation, with a significant improvement in quality of life.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16236841 B0012-3692 (Print) Clinical Trial Journal Article Multicenter Study16236841Department of Thoracic Medicine, Imperial College School of Medicine, National Heart and Lung Institute, Dovehouse St, London SW3 6LY, UK. ~?LMontuschi, P. Martello, S. Felli, M. Mondino, C. Barnes, P. J. Chiarotti, M.2005`Liquid chromatography/mass spectrometry analysis of exhaled leukotriene B4 in asthmatic children119 Respir Res6Oct 19 BACKGROUND: The role of leukotriene (LT) B4, a potent inflammatory mediator, in atopic asthmatic and atopic nonasthmatic children is largely unknown. The lack of a gold standard technique for measuring LTB4 in exhaled breath condensate (EBC) has hampered its quantitative assessment in this biological fluid. We sought to measure LTB4 in EBC in atopic asthmatic children and atopic nonasthmatic children. Exhaled nitric oxide (NO) was measured as an independent marker of airway inflammation. METHODS: Fifteen healthy children, 20 atopic nonasthmatic children, 25 steroid-naive atopic asthmatic children, and 22 atopic asthmatic children receiving inhaled corticosteroids were studied. The study design was of cross-sectional type. Exhaled LTB4 concentrations were measured using liquid chromatography/mass spectrometry-mass spectrometry (LC/MS/MS) with a triple quadrupole mass spectrometer. Exhaled NO was measured by chemiluminescence with a single breath on-line method. LTB4 values were expressed as the total amount (in pg) of eicosanoid expired in the 15-minute breath test. Kruskal-Wallis test was used to compare groups. RESULTS: Compared with healthy children [87.5 (82.5-102.5) pg, median and interquartile range], exhaled LTB4 was increased in steroid-naive atopic asthmatic [255.1 (175.0-314.7) pg, p < 0.001], but not in atopic nonasthmatic children [96.5 (87.3-102.5) pg, p = 0.59)]. Asthmatic children who were receiving inhaled corticosteroids had lower concentrations of exhaled LTB4 than steroid-naive asthmatics [125.0 (25.0-245.0) pg vs 255.1 (175.0-314.7) pg, p < 0.01, respectively]. Exhaled NO was higher in atopic nonasthmatic children [16.2 (13.5-22.4) ppb, p < 0.05] and, to a greater extent, in atopic steroid-naive asthmatic children [37.0 (31.7-57.6) ppb, p < 0.001] than in healthy children [8.3 (6.1-9.9) ppb]. Compared with steroid-naive asthmatic children, exhaled NO levels were reduced in asthmatic children who were receiving inhaled corticosteroids [15.9 (11.5-31.7) ppb, p < 0.01]. CONCLUSION: In contrast to exhaled NO concentrations, exhaled LTB4 values are selectively elevated in steroid-naive atopic asthmatic children, but not in atopic nonasthmatic children. Although placebo control studies are warranted, inhaled corticosteroids seem to reduce exhaled LTB4 in asthmatic children. LC/MS/MS analysis of exhaled LTB4 might provide a non-invasive, sensitive, and quantitative method for airway inflammation assessment in asthmatic children.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16236169 &1465-993X (Electronic) Journal Article16236169Department of Pharmacology, Faculty of Medicine, Catholic University of the Sacred Heart, Rome, Italy. pmontuschi@rm.unicatt.it ~?qBodini, A. D'Orazio, C. Peroni, D. Corradi, M. Folesani, G. Baraldi, E. Assael, B. M. Boner, A. Piacentini, G. L.2005sBiomarkers of neutrophilic inflammation in exhaled air of cystic fibrosis children with bacterial airway infections494-9Pediatr Pulmonol406Dec Leukotriene B(4) (LTB(4)) and interleukin-8 (IL-8) are inflammatory mediators involved in the neutrophil response to pulmonary bacterial colonization in cystic fibrosis (CF). The aim of this study was to investigate whether the LTB(4) and IL-8 levels in exhaled breath condensate (EBC) could be related to the type of bacterial colonization in CF patients. The pH level in EBC was analyzed as an estimate of airway acidification. Forty children were evaluated: 10 CF patients with P. aeruginosa, 10 CF patients with S. aureus, 10 not colonized CF patients, and 10 healthy children. LTB(4) and IL-8 in EBC were analyzed by specific enzyme immunoassay kits (EIA). The pH of EBC was measured with a pH-meter after deareation by bubbling with argon. Exhaled LTB(4) was higher in CF children with P. aeruginosa compared to those with S. aureus (P < 0.01), not colonized (P < 0.001), and healthy children (P < 0.01). Exhaled IL-8 was elevated in CF patients colonized by P. aeruginosa compared with other subgroups (vs. not colonized, P < 0.05; vs. healthy children, P < 0.001). IL-8 levels were higher in CF children with S. aureus than in healthy children (P < 0.05). There was an increase in IL-8 levels in not colonized CF patients compared with healthy children (P < 0.05). EBC pH was higher in healthy children compared to CF patients not colonized (P < 0.05). Our data suggest that EBC is suitable for evaluating neutrophil inflammatory mediators (LTB(4), IL-8, and pH) involved in the response to pulmonary bacterial colonization in CF children.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16229003 !8755-6863 (Print) Journal Article16229003>Department of Pediatrics, University of Verona, Verona, Italy.~?VNguyen, T. A. Woo-Park, J. Hess, M. Goins, M. Urban, P. Vaughan, J. Smith, A. Hunt, J.2005aAssaying all of the nitrogen oxides in breath modifies the interpretation of exhaled nitric oxide379-84Vascul Pharmacol436Adolescent Adult Asthma/diagnosis/*metabolism *Breath Tests Child Child, Preschool Female Humans Lung/physiopathology Male Middle Aged Nitric Oxide/*analysis Nitrogen Oxides/*analysis Research Support, N.I.H., Extramural Respiratory Function TestsDecExhaled nitric oxide (NO) assays measure the quantity of NO that emanates from the airway, not the amount of NO that is formed. Consumptive processes-including oxidation reactions-decrease the amount of gas phase NO available for exhalation. Higher oxides of nitrogen (HiNO(x)) are resulting reaction products, and are easily measured in exhaled breath condensate (EBC). We performed concurrent sampling of exhaled breath for gas phase NO and EBC HiNO(x) in controls and stable asthmatics. We identified that, mole for mole, asthma patients hourly exhale more HiNO(x) than they do NO, with a HiNO(x)/NO ratio of 1.21 (0.54-3.4). This is the reverse of the ratio found in controls, in whom the HiNO(x)/NO ratio was 0.75 (0.44-0.93), p=0.04. The sum of the hourly molar exhalation of NO and HiNO(x) was significantly higher in asthmatics (333 nmol/h (221-543) than controls (179 (138-231), p<0.001). We conclude that exhaled oxides of nitrogen are more informative when measured together as opposed to in isolation. We suggest that inflammation can be better evaluated with HiNO(x) and NO measured concurrently, and that the level of oxidation in the lung can be evaluated by comparing the easily measured ratios of HiNO(x) to NO in the exhaled breath.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16216561 01537-1891 (Print) Clinical Trial Journal Article16216561Division of Pediatric Respiratory Medicine, Asthma and Allergic Diseases Center, Box 800386, University of Virginia, Charlottesville, VA 22908, USA. ~?CKo, F. W. Lau, C. Y. Leung, T. F. Wong, G. W. Lam, C. W. Hui, D. S.2006Exhaled breath condensate levels of 8-isoprostane, growth related oncogene alpha and monocyte chemoattractant protein-1 in patients with chronic obstructive pulmonary disease630-8 Respir Med1004AprChronic obstructive pulmonary disease (COPD) patients have increased neutrophils and macrophages in their lungs, and inflammation of the airway is related to oxidative stress. This study assessed the levels of 8-isoprostane (an oxidative stress marker) and chemokines related to neutrophil and monocyte inflammation (growth-related oncogene alpha [GROalpha] and monocyte chemoattractant protein-1 [MCP-1]) in the airway of ex-smoking COPD patients by exhaled breath condensate (EBC) collection. Thirty-two (28 males) stable COPD patients (14 with FEV(1) 50% [Group 1], 18 with FEV(1) <50% predicted [Group 2]) and 18 non-smoking age and sex-matched controls were studied in this cross-sectional study. EBC was collected using the EcoScreen (Jaeger, Germany) during 10min of tidal breathing with the nose clipped. Concentrations of 8-isoprostane, GROalpha and MCP-1 were measured by enzyme immunoassays. COPD patients had a higher concentration of 8-isoprostane than controls (COPD versus control, P<0.001; Group 1 versus Group 2, P=0.045). 8-isoprostane increased across the groups from normal, Group 1 to Group 2 (r=0.64, P<0.001). The median intraquartile range (IQR) levels in pg/ml for GROalpha were 45.3(44.5-46.5), 45.4(44.5-46.0), 46.0(45.6-47.3), whereas MCP-1 levels were 5.3(5.2-5.9), 6.2(5.4-6.9) and 5.7(5.5-6.4) in Group 1, Group 2 COPD and control subjects, respectively. GROalpha level was lower in COPD patients when compared to controls (P=0.01). MCP-1 level did not differ between COPD and the control group. 8-isoprostane level, but not GROalpha and MCP-1, in EBC was increased in COPD patients with poorer lung function. This suggests an increased oxidative stress in the airway in patients with more severe COPD.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16213701 !0954-6111 (Print) Journal Article16213701Department of Medicine and Therapeutics, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, New Territories, Hong Kong. ~?9Davidsson, A. Naidu Sjosward, K. Lundman, L. Schmekel, B.2005RQuantitative assessment and repeatability of chlorine in exhaled breath condensate529-36 Respiration725Sep-Oct#BACKGROUND: Airway condition is presumably reflected in epithelial lining fluid (ELF). Exhaled breath condensate (EBC) has been used as a surrogate marker of the composition of ELF. OBJECTIVES: This study aimed at assessing the technical repeatability of chlorine measurements in EBC and comparing two separate condensators (Ecoscreen and R Tube regarding recovery and repeatability. Furthermore, the association between condensate recoveries and variations in the airway status were scrutinized. METHODS: EBC was collected using two condensators from 10 healthy volunteers. In addition, 13 asthmatic patients produced EBC with or without an added resistance of 5 cm H2O (Res5), applied to the outflow tract of Ecoscreen. All tests were done in random order. Chlorine levels (analyzed by a coulometric technique) in EBC served as a tool for investigation. RESULTS: Chlorine was measurable in all samples. The coefficient of repeatability of chlorine measurements was <10%. Chlorine levels were higher in EBC obtained from R Tube (p < 0.001), and differences in recoveries and variability in chlorine levels were presumably related to technical differences in the condensators and not to the repeatability of chlorine measurements per se. Air-flow-dependent chlorine levels were obtained from healthy volunteers. Application of Res5, recruiting additional alveoli, resulted in increased recovery of the EBC volume, but not of chlorine, from those that had the most pronounced airway obstruction (p = 0.05). CONCLUSION: We conclude that by employing a sensitive analysis technique, chlorine is repeatedly measurable in EBC. We suggest that the bulk of chlorine in EBC originates from large airways and not from the alveolar area. Both condensators were comparable regarding repeatability but differed regarding chlorine recovery.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16210893 !0025-7931 (Print) Journal Article16210893Department of Clinical Physiology, Institute of Medicine and Care, University Hospital, Linkoping, Sweden. anette.davidsson@lio.se~?cDeaton, C. M. Marlin, D. J. Smith, N. C. Harris, P. A. Dagleish, M. P. Schroter, R. C. Kelly, F. J.2005GEffect of acute airway inflammation on the pulmonary antioxidant status653-70 Exp Lung Res317Acute Disease Airway Obstruction/complications/physiopathology/veterinary Airway Resistance Animals Antioxidants/*metabolism Bronchoalveolar Lavage Fluid/chemistry/cytology Dust Female Horse Diseases/etiology/*metabolism/physiopathology Horses Leukocyte Elastase/metabolism Lung/*metabolism Male Neutrophils/pathology Oxidative Stress Pneumonia/etiology/*metabolism/physiopathology/*veterinary Research Support, Non-U.S. Gov'tSepEffects of acute airway inflammation induced by organic dust inhalation on pulmonary antioxidant status were investigated in healthy horses and horses affected by recurrent airway obstruction. Exposure to organic dust induced acute airway neutrophilia, which was associated with increases in elastase and decreases in ascorbic acid concentrations in bronchoalveolar lavage fluid. However, markers of oxidative stress were unaffected, as was hydrogen peroxide in breath condensate. Decreases in ascorbic acid correlated with increased respiratory resistance (P = 001) when both groups were combined. In conclusion, acute neutrophilic airway inflammation does not result in significant evidence of oxidative stress in horses affected by recurrent airway obstruction.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16203621 !0190-2148 (Print) Journal Article16203621mCentre for Equine Studies, Animal Health Trust, Lanwades Park, Kentford, Suffolk, UK. chris.deaton@aht.org.uk~?Wyse, C. A. Skeldon, K. Hotchkiss, J. W. Gibson, G. Yam, P. S. Christley, R. M. Preston, T. Cumming, D. R. Padgett, M. Cooper, J. C. Love, S.2005Effects of changes to the stable environment on the exhalation of ethane, carbon monoxide and hydrogen peroxide by horses with respiratory inflammation408-12Vet Rec15714"Airway Obstruction/epidemiology/metabolism/*veterinary Animals Breath Tests Carbon Monoxide/*analysis Cross-Over Studies Dust Environment Ethane/*analysis Horse Diseases/epidemiology/*metabolism Horses *Housing, Animal Hydrogen Peroxide/*analysis Recurrence Research Support, Non-U.S. Gov'tOct 1The aim of this study was to assess the effects of changes to the stable environment on exhaled markers of respiratory inflammation in six horses with clinical histories of recurrent airway obstruction. The horses were maintained for two weeks under conventional stable management (straw bedding and hay) and for two weeks on a reduced-dust regimen (paper bedding and ensiled grass), in a crossover study design. Exhaled ethane and carbon monoxide (CO) and exhaled breath condensate hydrogen peroxide (H(2)O(2)) were measured every three days under each regimen. The presence of clinical signs of airway inflammation (nasal discharge and cough) was monitored daily. The reduced-dust regimen was associated with fewer clinical signs of airway inflammation than the conventional regimen. Exhaled ethane and CO were significantly lower on the reduced-dust regimen and these markers were correlated with clinical signs of respiratory inflammation, but exhaled H(2)O(2) was not affected by the management regimen.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16199775 L0042-4900 (Print) Clinical Trial Journal Article Randomized Controlled Trial16199775Division of Companion Animal Sciences, Institute of Comparative Medicine, University of Glasgow Veterinary School, Bearsden, Glasgow G61 1QH, Scotland, UK.~?SAraneda, O. F. Garcia, C. Lagos, N. Quiroga, G. Cajigal, J. Salazar, M. P. Behn, C.2005Lung oxidative stress as related to exercise and altitude. Lipid peroxidation evidence in exhaled breath condensate: a possible predictor of acute mountain sickness383-90Eur J Appl Physiol955-6Dec+Lung oxidative stress (OS) was explored in resting and in exercising subjects exposed to moderate and high altitude. Exhaled breath condensate (EBC) was collected under field conditions in male high-competition mountain bikers performing a maximal cycloergometric exercise at 670 m and at 2,160 m, as well as, in male soldiers climbing up to 6,125 m in Northern Chile. Malondialdehyde concentration [MDA] was measured by high-performance liquid chromatography in EBC and in serum samples. Hydrogen peroxide concentration [H(2)O(2)] was analysed in EBC according to the spectrophotometric FOX(2) assay. [MDA] in EBC of bikers did not change while exercising at 670 m, but increased from 30.0+/-8.0 to 50.0+/-11.0 nmol l(-1) (P<0.05) at 2,160 m. Concomitantly, [MDA] in serum and [H(2)O(2)] in EBC remained constant. On the other hand, in mountaineering soldiers, [H(2)O(2)] in EBC under resting conditions increased from 0.30+/-0.12 mumol l(-1) at 670 m to 1.14+/-0.29 mumol l(-1) immediately on return from the mountain. Three days later, [H(2)O(2)] in EBC (0.93 +/-0.23 mumol l(-1)) continued to be elevated (P<0.05). [MDA] in EBC increased from 71+/-16 nmol l(-1) at 670 m to 128+/-26 nmol l(-1) at 3,000 m (P<0.05). Changes of [H(2)O(2)] in EBC while ascending from 670 m up to 3,000 m inversely correlated with concomitant variations in HbO2 saturation (r=-0.48, P<0.05). AMS score evaluated at 5,000 m directly correlated with changes of [MDA] in EBC occurring while the subjects moved from 670 to 3,000 m (r=0.51, P<0.05). Lung OS may constitute a pathogenic factor in AMS.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16195882 !1439-6319 (Print) Journal Article16195882Laboratorio de Ambientes Extremos, Programa de Fisiologia y Biofisica, ICBM, Facultad de Medicina, Universidad de Chile, Santiago, Chile. oaraneda@med.uchile.cl~? JKwiatkowska, S. Luczynska, M. Grzelewska-Rzymowska, I. Nowak, D. Zieba, M.2005[Comparison of oxidative stress markers in exhaled breath condensate and in serum of patients with tuberculosis and sarcoidosis]37-40Pol Merkuriusz Lek19109 Adult Biological Markers/metabolism *Breath Tests Case-Control Studies English Abstract Exhalation Female Humans Hydrogen Peroxide/*metabolism *Lipid Peroxidation Male Middle Aged *Oxidative Stress Sarcoidosis/blood/*metabolism Tuberculosis, Pulmonary/blood/*metabolismJul?Tuberculosis and sarcoidosis represent the granulomatous diseases. The aim of the study was to compare the markers of oxidative stress: in exhaled breath condensate (EBC) and in serum of patients with tuberculosis and sarcoidosis. MATERIAL AND METHODS: 19 patients with active lung tuberculosis and 15 patients with sarcoidosis were enrolled into the study. As a control served 15 healthy subjects. Hydrogen peroxide (H2O2) was measured in EBC and the ends products of lipid peroxidation (TBARs) were assessed in serum. RESULTS: The concentrations of H202 and TBARs (1022.96+/-186.02 nM and 4.22+/-0. 80 microM, respectively) were significantly higher in patients with tuberculosis as compared with the controls (398.15+/-37.10 nM and 0.48+/-0.17 microM, respectively). The patients with sarcoidosis revealed only the significantly elevated levels of hydrogen peroxide (963.30+/-105.77 nM) in breath condensate. CONCLUSIONS: It was found that local and systemic oxidative stress were present in patients with tuberculosis, while in those with sarcoidosis existed only the local reaction.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16194024 !1426-9686 (Print) Journal Article16194024Porownanie wybranych wykladnikow stresu oksydacyjnego w kondensacie powietrza wydechowego oraz w surowicy u chorych na gruzlice i sarkoidoze.*Uniwersytet Medyczny w Lodzi. skwiat@wp.pl v~?!xMontuschi, P. Macagno, F. Parente, P. Valente, S. Lauriola, L. Ciappi, G. Kharitonov, S. A. Barnes, P. J. Ciabattoni, G.2005REffects of cyclo-oxygenase inhibition on exhaled eicosanoids in patients with COPD827-33Thorax6010Blood Gas Analysis/methods Cross-Over Studies Cyclooxygenase Inhibitors/*therapeutic use Dinoprostone/metabolism Double-Blind Method Eicosanoids/*metabolism Female Forced Expiratory Volume/physiology Humans Ibuprofen/*therapeutic use Lactones/*therapeutic use Leukotriene B4/metabolism Male Middle Aged Pulmonary Disease, Chronic Obstructive/*drug therapy/metabolism Research Support, Non-U.S. Gov't Sputum/chemistry Sulfones/*therapeutic use Thromboxane B2/metabolism Vital Capacity/physiologyOctBACKGROUND: Leukotriene (LT) B4 concentrations are increased and prostaglandin (PG) E2 concentrations are decreased in exhaled breath condensate (EBC) in patients with chronic obstructive pulmonary disease (COPD). A study was undertaken to investigate the short term effects of cyclo-oxygenase (COX) inhibition on exhaled LTB4 and PGE2 concentrations in patients with COPD and to identify the COX isoform responsible for exhaled PGE2 production. METHODS: Two studies were performed. A double blind, crossover, randomised, placebo controlled study with ibuprofen (400 mg qid for 2 days), a non-selective COX inhibitor, was undertaken in 14 patients with stable COPD, and an open label study with oral rofecoxib (25 mg once a day for 5 days), a selective COX-2 inhibitor, was undertaken in a different group of 16 COPD patients. EBC was collected before and after drug treatment. Exhaled LTB4 and PGE2 concentrations were measured with specific immunoassays. RESULTS: All patients complied with treatment as indicated by a reduction in ex vivo serum thromboxane B2 concentrations (ibuprofen) and a reduction in lipopolysaccharide induced increase in ex vivo plasma PGE2 values (rofecoxib) of more than 80%. Exhaled LTB4 was increased after ibuprofen (median 175.5 (interquartile range 128.8-231.5) pg/ml v 84.0 (70.0-98.5) pg/ml, p < 0.001) and exhaled PGE2 was reduced (93.5 (84.0-105-5) pg/ml v 22.0 (15.0-25.5) pg/ml, p < 0.0001). Rofecoxib had no effect on exhaled LTB4 (p = 0.53) or PGE2 (p = 0.23). CONCLUSIONS: Non-selective COX inhibition decreases PGE2 and increases LTB4 in EBC, whereas selective COX-2 inhibition has no effect on these eicosanoids. PGE2 in EBC is primarily derived from COX-1 activity, and COX inhibition may redirect arachidonic acid metabolism towards the 5-lipoxygenase pathway.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16192367 L0040-6376 (Print) Clinical Trial Journal Article Randomized Controlled Trial16192367Department of Pharmacology, Faculty of Medicine, School of Medicine, Catholic University of the Sacred Heart, Largo F Vito 1, 00168 Rome, Italy. pmontuschi@rm.unicatt.itq~?"(Simpson, J. L. Wood, L. G. Gibson, P. G.2005CInflammatory mediators in exhaled breath, induced sputum and saliva1180-5Clin Exp Allergy359^Adult Aged Asthma/*immunology Breath Tests/methods Bronchial Provocation Tests Dinoprost/*analogs & derivatives/analysis Female Humans Interleukin-8/*analysis Male Middle Aged Predictive Value of Tests Pulmonary Surfactant-Associated Protein A/*analysis Research Support, Non-U.S. Gov't Saliva/*immunology Sputum/*immunology Statistics, NonparametricSepIBACKGROUND AND OBJECTIVE: Airway inflammation is assessed to monitor progression, control and treatment of asthma. The collection of exhaled breath condensate (EBC) provides a non-invasive alternative to induced sputum samples for the monitoring of airway inflammation. Both samples can be confounded by salivary contamination. The aim of this study was to compare the levels of inflammatory mediators in samples of EBC, induced sputum and saliva samples from subjects with asthma. METHOD: EBC, saliva and induced sputum samples were collected from subjects with asthma (n=10). Total protein, IL-8, 8-isoprostane and surfactant protein A (SPA) were assessed in each sample. RESULTS: Total protein, IL-8, 8-isoprostane and SPA were detected in all sputum samples. Only total protein and SPA were consistently measured in EBC, with levels at least 100-fold lower than those measured in induced sputum. In saliva, total protein, SPA and 8-isoprostane were detected in all samples, with IL-8 detected in 60% of samples. CONCLUSIONS: Induced sputum is a reliable technique that can be used to assess markers of airway inflammation. While EBC is a simple and inexpensive technique to collect lower airway secretions, the detection of inflammatory mediators is variable, and further work is required to validate this technique to assess inflammatory mediators.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16164445 40954-7894 (Print) Evaluation Studies Journal Article16164445hSchool of Medical Practice and Population Health, The University of Newcastle, Callaghan NSW, Australia.n~?#BHuszar, E. Szabo, Z. Jakab, A. Barta, I. Herjavecz, I. Horvath, I.2005lComparative measurement of thromboxane A2 metabolites in exhaled breath condensate by different immunoassays350-5 Inflamm Res548Adult Aged Asthma/metabolism Breath Tests Bronchi/pathology Comparative Study Female Humans Immunoassay/*methods Immunoenzyme Techniques Male Middle Aged Radioimmunoassay Research Support, Non-U.S. Gov't Respiratory Function Tests Thromboxane A2/*metabolismAugjOBJECTIVE: Differences between detection techniques may be partly responsible for variable mediator concentrations reported in exhaled breath condensate (EBC). We compared two types of immunoassays to estimate thromboxane A(2) (TxA(2)) concentration. MATERIALS AND METHODS: Thromboxane B(2) (TxB(2)) levels were measured by enzyme immunoassay (EIA) and TxB(2)/2,3-dinor TxB(2) by radioimmunoassay (RIA) in 10 healthy subjects and 13 asthmatic patients. 2,3-Dinor TxB(2) was also determined by a separate EIA. RESULTS: Thromboxane was detected in all samples by RIA, but only in about 75% of samples by EIA. 2,3-Dinor TxB(2) was detected in most samples. There was no agreement between the results of the different immunoassays. As compared to healthy subjects, exhaled breath condensates of asthmatic patients contained significantly more immunoreactivity by RIA and TxB(2) EIA (but not by 2,3-dinor TxB(2) EIA). CONCLUSION: RIA and EIA resulted in vastly different absolute values. The difference found between healthy volunteers and asthmatic patients however, suggests an increased level of TxA(2) in the airways of asthmatics.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16158336 !1023-3830 (Print) Journal Article16158336wDepartment of Pathophysiology, National Koranyi Institute for TB and Pulmonology, Budapest, Hungary. hildiko@koranyi.hu h~?$6Goen, T. Muller-Lux, A. Dewes, P. Musiol, A. Kraus, T.2005`Sensitive and accurate analyses of free 3-nitrotyrosine in exhaled breath condensate by LC-MS/MS261-6-J Chromatogr B Analyt Technol Biomed Life Sci8261-2Adult Biological Markers/analysis Breath Tests/*methods Chromatography, Liquid/methods *Exhalation Humans Sensitivity and Specificity Smoking Spectrometry, Mass, Electrospray Ionization/methods Tyrosine/*analogs & derivatives/analysisNov 5TThe quantitative determination of 3-nitro-l-tyrosine, a biological marker for inflammatory processes, in exhaled breath condensate (EBC) is described. The clean-up and preconcentration was performed by solid phase extraction (SPE). After liquid chromatography the specific detection was performed by tandem mass spectrometry using electron spray ionisation and selected reaction monitoring (SRM). 13C9-3-nitrotyrosine was used as an internal standard. For reliability, tests for the precision of the method, the losses during preparation, a test for nitrating artifacts and the comparibility of calibrants in EBC and buffer solution were performed. The calibration of the method was linear over a range of 10-500 pg/mL. The within-run coefficients of variation (CV) of the samples were found to be 8.4% at 25 pg/mL and 8.3% at 250 pg/mL. The day-to-day CV was found to be 11.2%. The limit of quantification was 3.9 pg/mL. The losses during preparation were 15%. The discrepancy between the calibration with EBC and buffer solution was below 10%. No artificial production of 3-nitrotyrosine was observed during the procedure. The application of the method on the EBC samples of healthy smokers (N=10) and non-smokers (N=10) showed no difference between the two groups. The concentration of 3-nitrotyrosine ranged between the limit of quantification and 184 pg/mL and was distinctly lower than data detected by an immunoassay procedure. The procedure was proven to be accurate, sensitive and in contrast to GC methods less elaborate and is recommended for the determination of 3-nitrotyrosine in exhaled breath condensate.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16143570 !1570-0232 (Print) Journal Article16143570Institute for Occupational Medicine and Social Medicine, RWTH Aachen, Pauwelsstr. 30, D-52074 Aachen, Germany. Thomas.Goeen@post.rwth-aachen.de 0F~?%GChladkova, J. Krcmova, I. Chladek, J. Cap, P. Micuda, S. Hanzalkova, Y.2005KValidation of Nitrite and Nitrate Measurements in Exhaled Breath Condensate RespirationAug 30dBackground: Inflammatory markers in exhaled breath condensate (EBC) are investigated as a non-invasive approach to monitoring of inflammation in the respiratory tract. EBC concentrations of nitrite and nitrate, the stable end products of oxidative metabolism of nitric oxide, are increased in patients with asthma, especially during acute exacerbations. Objectives: To examine methodological aspects of nitrite and nitrate measurements in EBC such as sample collection, storage and analysis. Methods: In a randomized study, EBC was collected twice within 1 h (with and without a nose clip) in 20 healthy adults and 20 patients with well-controlled asthma and no symptoms of allergic rhinitis. Nitrite and nitrate were assayed by ionex chromatography and fluorimetrically after derivatization with diaminonaphthalene. Results: The geometric mean [exp (mean +/- SD)] EBC levels of nitrite and nitrate in healthy subjects [4.3 (3.0-6.1) and 11.0 (5.3-22.7) mumo/l] and patients [4.6 (2.6-7.3) and 8.7 (3.2-23.8) mumo/l] did not differ (p = 0.13). Wearing a nose clip (p = 0.3) did not influence nitrite and nitrate concentrations. The mean intra-subject %CVs of EBC concentrations of nitrite were 26 and 21% in healthy subjects and patients, while those of nitrate achieved 49 and 88%, respectively. Conclusions: Ionex chromatography of nitrite and nitrate requires no sample pretreatment and provides comparable results as a more laborious diaminonaphthalene method. EBC samples should be kept cold (8 degrees C) and analyzed for nitrite and nitrate within 24 h of collection or stored in the freezer and thawed preferably only once. Wearing a nose clip during EBC collection has no influence on nitrite and nitrate concentrations. Short-term repeatability of nitrite and nitrate measurements was worse compared to published data on exhaled nitric oxide. Copyright (c) 2005 S. Karger AG, Basel.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16141708 !0025-7931 (Print) Journal article16141708mDepartment of Pediatrics,Charles University in Prague, Faculty of Medicine in Hradec Kralove, Czech Republic.~?&yGessner, C. Scheibe, R. Wotzel, M. Hammerschmidt, S. Kuhn, H. Engelmann, L. Hoheisel, G. Gillissen, A. Sack, U. Wirtz, H.2005TExhaled breath condensate cytokine patterns in chronic obstructive pulmonary disease1229-40 Respir Med9910OctDifferences 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.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16140223 !0954-6111 (Print) Journal Article16140223vDepartment of Respiratory Medicine, University of Leipzig, Johannisallee 32, 04103 Leipzig, Germany. ch.gessner@web.de1~?'^Goldoni, M. Caglieri, A. Andreoli, R. Poli, D. Manini, P. Vettori, M. V. Corradi, M. Mutti, A.2005KInfluence of condensation temperature on selected exhaled breath parameters10 BMC Pulm Med5Sep 1(BACKGROUND: The effects of changes in cooling temperature on biomarker levels in exhaled breath condensate have been little investigated. The aim of the study was to test the effect of condensation temperature on the parameters of exhaled breath condensate and the levels of selected biomarkers. METHODS: Exhaled breath condensate was collected from 24 healthy subjects at temperatures of -10, -5, 0 and +5 C degrees. Selected parameters (condensed volume and conductivity) and biomarkers (hydrogen peroxide, malondialdehyde) were measured. RESULTS: There was a progressive increase in hydrogen peroxide and malondialdehyde concentrations, and condensate conductivity as the cooling temperature increased; total condensate volume increased as the cooling temperature decreased. CONCLUSION: The cooling temperature of exhaled breath condensate collection influenced selected biomarkers and potential normalizing factors (particularly conductivity) in different ways ex vivo. The temperature of exhaled breath condensate collection should be controlled and reported.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16137323 &1471-2466 (Electronic) Journal Article16137323National Institute of Occupational Safety and Prevention, Research Center at the University of Parma, Parma, Italy. matgold@libero.it ~?(Horvath, I. Hunt, J. Barnes, P. J. Alving, K. Antczak, A. Baraldi, E. Becher, G. van Beurden, W. J. Corradi, M. Dekhuijzen, R. Dweik, R. A. Dwyer, T. Effros, R. Erzurum, S. Gaston, B. Gessner, C. Greening, A. Ho, L. P. Hohlfeld, J. Jobsis, Q. Laskowski, D. Loukides, S. Marlin, D. Montuschi, P. Olin, A. C. Redington, A. E. Reinhold, P. van Rensen, E. L. Rubinstein, I. Silkoff, P. Toren, K. Vass, G. Vogelberg, C. Wirtz, H.2005RExhaled breath condensate: methodological recommendations and unresolved questions523-48 Eur Respir J263Biological Markers/metabolism Breath Tests/*methods Humans Lung Diseases/diagnosis/*metabolism Oxidative Stress/physiology Reproducibility of ResultsSepCollection 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.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16135737 >0903-1936 (Print) Guideline Journal Article Practice Guideline16135737National Koranyi Institute for Pulmonology, Dept of Pathophysiology, Budapest PO Box 1, Piheno u. 1. H-1529, Hungary. hildiko@koranyi.hu~?)Holz, O.2005OCatching breath: monitoring airway inflammation using exhaled breath condensate371-2 Eur Respir J263Biological Markers/metabolism *Breath Tests Humans Inflammation/metabolism Lung Diseases, Obstructive/*metabolism Practice GuidelinesSepfhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16135713 #0903-1936 (Print) Comment Editorial16135713 ~?*lCarpagnano, G. E. Foschino Barbaro, M. P. Resta, O. Gramiccioni, E. Valerio, N. V. Bracciale, P. Valerio, G.2005{Exhaled markers in the monitoring of airways inflammation and its response to steroid's treatment in mild persistent asthma175-81Eur J Pharmacol5191-2Administration, Inhalation Adult Androstadienes/administration & dosage/*therapeutic use Anti-Inflammatory Agents/administration & dosage/*therapeutic use Asthma/*drug therapy/metabolism/physiopathology Biological Markers/analysis Breath Tests Comparative Study Female Humans Hydrogen-Ion Concentration Interleukin-4/*analysis Interleukin-6/*analysis Male Middle Aged Monitoring, Physiologic/methods Reproducibility of ResultsSep 5BThe measure of inflammatory cytokines in the exhaled breath condensate has been recently proposed for use in monitoring asthma and the therapeutic response to steroids. The aim of the present study was to investigate the usefulness of measuring exhaled IL-6, IL-4 and pH in mild persistent asthma. Furthermore the effects on these markers of inhaled steroids were assessed. The study enrolled 28 asthmatic (15 males, 38+/-12 years) and 15 healthy subjects (5 males, 35+/-6 years). IL-6, IL-4 and pH were measured in the exhaled breath condensate of the subjects studied. Significantly higher concentrations of IL-6 and IL-4 were observed in the breath condensate of asthmatic patients (7.1+/-1.1 and 64.4+/-8.3 pg/ml) compared to controls (2.7+/-0.6 and 31.7+/-3.5 pg/ml), p<0.001. Furthermore, exhaled IL-4 fell significantly after treatment with inhaled steroids for 6 months (47.9+/-3.2 pg/ml, p<0.001) while exhaled IL-6 did not (6.4+/-1.0 pg/ml, p=0.8). The exhaled pH turned out to be lower in asthmatic subjects than in controls (7.39+/-0.11 vs. 7.85+/-0.14; P<0.001) but trended towards control levels after steroid treatment (7.65+/-0.16, P<0.001). We conclude that the measurement of exhaled IL-4 and pH in mild asthmatic subjects could be a useful way of monitoring their airway inflammation as well as their response to the treatment.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16122732 00014-2999 (Print) Clinical Trial Journal Article16122732UInstitute of Respiratory Disease, University of Foggia, Italy. ge.carpagnano@unifg.itO~?+ Gustin, P.2005N[Development of a model of asthma in the cat: veterinary and medical concerns]133-9; discussion 140Bull Mem Acad R Med Belg1601-2}Animals Asthma/therapy/*veterinary Cat Diseases/*physiopathology/*therapy Cats Disease Models, Animal English Abstract Humans/Feline asthma is a respiratory disorder, which is poorly studied in veterinary medicine and whose therapeutical approach is not based on pre-clinical or clinical studies. Besides the need to establish therapeutical strategies using bronchodilators and anti-inflammatory drugs, the asthmatic cat presents some interest as animal model of human asthma. Given the body size of a cat and the similarities of pathophysiology between feline and human asthma, simultaneous and repeated investigations of lung function, airway inflammation and airway remodelling in response to an allergic stimulus might be performed in felines. The experimental model of feline asthma developed by our laboratory allows, by respecting animal well-being, (1) the non invasive investigation of lung function and airway responsiveness using barometric whole body plethysmography, (2) the investigation of lower airway inflammation by determining hydrogen peroxide in exhaled breath condensate and by analysing bronchoalveolar lavage fluid, (3) the assessment of morphological changes based on thoracic radiography, bronchoscopic scoring and bronchial biopsies. This experimental model allows, on one hand, to elaborate therapeutical approaches for spontaneously asthmatic cats and, on the other hand, the testing of innovative treatments against asthma.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16116817 !0377-8231 (Print) Journal Article16116817SDeveloppement d'un modele d'asthme chez le chat: interet veterinaire et biomedical. u~?,Kharitonov, S. A. Barnes, P. J.2004xEffects of corticosteroids on noninvasive biomarkers of inflammation in asthma and chronic obstructive pulmonary disease191-9Proc Am Thorac Soc13NAdministration, Inhalation Adrenal Cortex Hormones/*administration & dosage Asthma/diagnosis/*drug therapy Biological Markers/*analysis Breath Tests Carbon Monoxide/metabolism Case-Control Studies Comparative Study Dose-Response Relationship, Drug Drug Administration Schedule Enzyme-Linked Immunosorbent Assay Female Humans Male Nitric Oxide/metabolism Prognosis Proteomics Pulmonary Disease, Chronic Obstructive/diagnosis/*drug therapy Pulmonary Gas Exchange Research Support, Non-U.S. Gov't Respiratory Function Tests Risk Assessment Sensitivity and Specificity Severity of Illness IndexExhaled breath analysis may be used to quantify inflammation and oxidative stress in the respiratory tract, in the differential diagnosis of airway diseases, and in the monitoring of therapy. The greatest progress has been made with standardized measurement of exhaled nitric oxide (NO). Bronchial NO is increased in asthma, correlated with other markers of inflammation, and reduced by treatment with corticosteroids and antileukotrienes. Alveolar NO is increased in chronic obstructive pulmonary disease (COPD), reflecting disease severity and progression. Exhaled carbon monoxide and ethane are increased in both asthma and COPD. Increased concentrations of 8-isoprostane, hydrogen peroxide, nitrite, and nitrotyrosine are found in exhaled breath condensate from patients with inflammatory lung diseases. Increased levels of lipid mediators are also found, and the pattern depends on the nature of the disease process. Additional biomarkers are exhaled breath temperature, which is elevated in asthma and reduced in COPD, and bronchial blood flow. It is likely that smaller and more sensitive analyzers will extend the discriminatory value of exhaled breath analysis and that new techniques will become available to diagnose and monitor respiratory diseases in the family practice and home settings.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16113434 (1546-3222 (Print) Journal Article Review16113434Department of Thoracic Medicine, National Heart & Lung Institute, Imperial College, Dovehouse Street, London SW3 6LY, UK. s.kharitonov@imperial.ac.uk~?-JGerritsen, W. B. Zanen, P. Bauwens, A. A. van den Bosch, J. M. Haas, F. J.2005TValidation of a new method to measure hydrogen peroxide in exhaled breath condensate1132-7 Respir Med999.Adult Biological Markers/analysis Biosensing Techniques Breath Tests/instrumentation/*methods Exhalation Female Humans Hydrogen Peroxide/*analysis Male Middle Aged Oxidative Stress Pulmonary Disease, Chronic Obstructive/diagnosis Reproducibility of Results Smoking/metabolism Spectrometry, FluorescenceSephInflammatory processes in the lung can or will elicit oxidative stress. The degree of oxidative stress can be determined by measuring hydrogen peroxide (H(2)O(2)) concentration in exhaled breath condensate (EBC) but the methods to measure H(2)O(2) are all rather time consuming and only reliable and/or accurate in the hand of skilled technicians in a dedicated laboratory. We tested a new commercial device (Ecocheck), developed to offer a less time-consuming method to measure H(2)O(2). We validated this new method according the NCCLS EP10-A2 protocol. The validation shows that the imprecision in the low range is high (28.4%) and declines with higher concentrations of H(2)O(2) (up to 6.6%). The Ecocheck is "an easy to use" measuring device for routine measurements getting quick results without the need for skilled technicians to determine H(2)O(2) concentrations.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16085214 40954-6111 (Print) Journal Article Validation Studies16085214Department of Clinical Chemistry, St. Antonius Hospital, PO Box 2500, NL-3430 EM Nieuwegein, The Netherlands. icb75097@nifty.com ~?.~Battaglia, S. den Hertog, H. Timmers, M. C. Lazeroms, S. P. Vignola, A. M. Rabe, K. F. Bellia, V. Hiemstra, P. S. Sterk, P. J.2005JSmall airways function and molecular markers in exhaled air in mild asthma639-44Thorax608BAdult Asthma/*physiopathology Biological Markers/analysis Bronchitis/*physiopathology Cross-Sectional Studies Dinoprost/*analogs & derivatives/analysis Enzyme-Linked Immunosorbent Assay Female Forced Expiratory Volume/physiology Humans Male Nitric Oxide/*analysis Research Support, Non-U.S. Gov't Vital Capacity/physiologyAugBACKGROUND: Several studies suggest that the periphery of the lung is the major site of inflammation in asthma. Fractional exhaled nitric oxide (Feno) and 8-isoprostane have been proposed as biomarkers of inflammation and oxidative stress. We therefore hypothesised that small airway dysfunction in asthma is of inflammatory origin that can be detected by molecular markers in exhaled air. To test this hypothesis, we examined the relationship of Feno and 8-isoprostane in exhaled air with small airways function as assessed by the single breath nitrogen test. METHODS: Sixteen patients (14 women) with mild atopic asthma (forced expiratory volume in 1 second >80% predicted) of mean (SD) age 23.0 (5.5) years participated in a cross sectional study. Feno was recorded by chemiluminescence and 8-isoprostane was measured by ELISA in concentrated exhaled breath condensate. The slope of phase III (deltaN2) and the closing volume (CV) were assessed from the single breath washout curve. RESULTS: The median Feno level was 30.4 ppb (range 10.1-82.8), the median 8-isoprostane concentration in exhaled breath condensate was 2.2 pg/ml (range 1.6-2.7), and the mean (SD) deltaN2 value was 1.1 (0.4)% N2/l. Feno was positively associated with deltaN2 (r(s) = 0.54, p = 0.032) while 8-isoprostane was inversely correlated with FEV1% predicted (rs= -0.58; p = 0.017) and CV as a percentage of vital capacity (rs= 0.58; p = 0.019). CONCLUSIONS: Feno and 8-isoprostane in exhaled air are associated with small airways function in mild asthma. This suggests that these markers reflect small airway inflammation and favours a role for them as disease markers that is complementary to spirometry in the monitoring of patients with asthma.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16061704 !0040-6376 (Print) Journal Article16061704yLung Function Laboratory C2-P, Leiden University Medical Centre (LUMC), P O Box 9600, NL-2300 RC Leiden, The Netherlands.~?/VMacGregor, G. Ellis, S. Andrews, J. Imrie, M. Innes, A. Greening, A. P. Cunningham, S.2005CBreath condensate ammonium is lower in children with chronic asthma271-6 Eur Respir J262AugfExhaled breath condensate pH and ammonium reflect asthmatic status and acute exacerbations in adults. The aim of this study was to assess whether pH and ammonium could reflect asthma and its severity in children. The current study comprised two parts: 1) a cross-section of 74 children with asthma (median age 10.5 yrs) compared with 47 healthy controls (median age 10 yrs); and 2) longitudinal assessment of eight children (mean age 8.5 yrs) admitted with asthma exacerbation. Condensate pH and ammonium were compared with clinical observations. In the cross-sectional part of the study, lower per cent forced expiratory volume in one second was associated with more symptoms and treatment. There was no significant difference between median pH in children with stable asthma (6.05) compared with controls (5.90). Ammonium was significantly lower in children with asthma (median 258 microM) compared with controls (median 428 microM). No association was found between ammonium or pH and lung function or symptom-free days. In the longitudinal study, significant improvements in oxygen saturation and respiratory rate with treatment of an acute exacerbation were not reflected by changes in pH or ammonium. In conclusion, pH does not appear to reflect disease or severity in children with asthma. Ammonium was significantly lower in children with asthma when compared with controls.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16055875 !0903-1936 (Print) Journal Article16055875CRespiratory Medicine Unit, Western General Hospital, Edinburgh, UK. ~?08Leung, T. F. Wong, G. W. Ko, F. W. Lam, C. W. Fok, T. F.2005eClinical and atopic parameters and airway inflammatory markers in childhood asthma: a factor analysis822-6Thorax6010Administration, Inhalation Adolescent Adrenal Cortex Hormones/administration & dosage Asthma/*diagnosis/physiopathology Biological Markers/blood Bronchitis/*diagnosis/physiopathology Chemokines/metabolism Child Chronic Disease Factor Analysis, Statistical Forced Expiratory Volume/physiology Humans Hypersensitivity, Immediate/*diagnosis/physiopathology Leukotriene B4/metabolism Nitric Oxide/metabolism Research Support, Non-U.S. Gov't Vital Capacity/physiologyOctBACKGROUND: Recent studies have repeatedly shown weak correlations among lung function parameters, atopy, exhaled nitric oxide level (Feno), and airway inflammatory markers, suggesting that they are non-overlapping characteristics of asthma in adults. A study was undertaken to determine, using factor analysis, whether the above features represent separate dimensions of childhood asthma. METHODS: Clinically stable asthmatic patients aged 7-18 years underwent spirometric testing, methacholine bronchial challenge, blood sampling for atopy markers and chemokine levels (macrophage derived chemokine (MDC), thymus and activation regulated chemokine (TARC), and eotaxin), Feno, and chemokines (MDC and eotaxin) and leukotriene B(4) measurements in exhaled breath condensate (EBC). RESULTS: The mean (SD) forced expiratory volume in 1 second (FEV1) and Feno of 92 patients were 92.1 (15.9)% predicted and 87.3 (65.7) ppb, respectively. 59% of patients received inhaled corticosteroids. Factor analysis selected four different factors, explaining 55.5% of total variance. The Kaiser-Meyer-Olkin measure of sampling adequacy was 0.587. Plasma total and specific IgE levels, peripheral blood eosinophil percentage, and Feno loaded on factor 1; plasma TARC and MDC concentrations on factor 2; MDC, eotaxin and leukotriene B4 concentrations in EBC on factor 3; and plasma eotaxin concentration together with clinical indices including body mass index and disease severity score loaded on factor 4. Post hoc factor analyses revealed similar results when outliers were excluded. CONCLUSIONS: The results suggest that atopy related indices and airway inflammation are separate dimensions in the assessment of childhood asthma, and inflammatory markers in peripheral blood and EBC are non-overlapping factors of asthma.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16055623 !0040-6376 (Print) Journal Article16055623Department of Paediatrics, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, NT, Hong Kong SAR, China. tfleung@cuhk.edu.hk~?1Liu, J. Thomas, P. S.2005gExhaled breath condensate as a method of sampling airway nitric oxide and other markers of inflammationMT53-62 Med Sci Monit118Animals Biological Markers/analysis Breath Tests/*methods *Exhalation Humans Inflammation/metabolism Nitric Oxide/*analysis/metabolism Research Support, Non-U.S. Gov'tAugDMost of the methods of investigating lung diseases have been invasive until the discovery that exhaled nitric oxide can be used as a surrogate marker of airway inflammation, particularly in asthma. Exhaled nitric oxide (NO) is now established as a marker of airway inflammation. It has been shown to correlate well with eosinophilic asthmatic airway inflammation, and to be able to predict decline in asthma control and airway function. Altered levels of NO are also associated with other inflammatory lung diseases. In addition, polymorphisms of the genes encoding the three nitric oxide synthases are associated with phenotypic differences associated with lung diseases. Exhaled NO is, however, non-specific. It is therefore of importance that collecting exhaled breath condensate (EBC) has emerged as a potential tool in the study of pulmonary diseases. The exhaled breath is collected in a cooling system which allows water vapour to condense. The EBC contains a number of mediators relating to the NO pathway, including nitrite as a metabolite of nitric oxide, nitrotyrosine, nitrosothiols plus small molecular mediators associated with oxidative stress, including hydrogen ions, and hydrogen peroxide. In addition, reports are emerging of the detection of larger molecules which not only include leukotrienes, prostaglandins, albumin and other proteins, such as cytokines, but also macromolecules, for example, DNA. EBC is becoming a technique which will allow repeated non-invasive sampling from the respiratory tract thus assisting pulmonary research and possibly the monitoring of lung diseases.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16049390 (1234-1010 (Print) Journal Article Review16049390Faculty of Medicine, University of New South Wales and Department of Respiratory Medicine, Prince of Wales Hospital, Randwick, NSW, Australia. ~?2DShahid, S. K. Kharitonov, S. A. Wilson, N. M. Bush, A. Barnes, P. J.2005)Exhaled 8-isoprostane in childhood asthma79 Respir Res6Jul 21kBACKGROUND: Exhaled breath condensate (EBC) is a non-invasive method to assess airway inflammation and oxidative stress and may be useful in the assessment of childhood asthma. METHODS: Exhaled 8-isoprostane, a stable marker of oxidative stress, was measured in EBC, in children (5-17 years) with asthma (13 steroid-naive and 12 inhaled steroid-treated) and 11 healthy control. RESULTS: Mean exhaled 8-isoprostane concentration was significantly elevated in steroid-naive asthmatic children compared to healthy children 9.3 (SEM 1.7) vs. 3.8 (0.6) pg/ml, p < 0.01. Children on inhaled steroids also had significantly higher 8-isoprostane levels than those of normal subjects 6.7 (0.7) vs. 3.8 (0.6) pg/ml, p < 0.01. Steroid-naive asthmatics had higher exhaled nitric oxide (eNO) than those of controls 28.5 (4.7) vs. 12.6 (1.5) ppb, p < 0.01. eNO in steroid-treated asthmatics was similar to control subjects 27.5(8.8) vs. 12.6(1.5) ppb. Exhaled 8-isoprostane did not correlate with duration of asthma, dose of inhaled steroids or eNO. CONCLUSION: We conclude that 8-isoprostane is elevated in asthmatic children, indicating increased oxidative stress, and that this does not appear to be normalized by inhaled steroid therapy. This suggests that 8-isoprostane is a useful non-invasive measurement of oxidative stress in children and that antioxidant therapy may be useful in the future.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16042771 &1465-993X (Electronic) Journal Article16042771WNational Heart and Lung Institute, Imperial College London, London, UK. a.bush@ic.ac.uk ~?3TMercken, E. M. Hageman, G. J. Schols, A. M. Akkermans, M. A. Bast, A. Wouters, E. F.2005cRehabilitation decreases exercise-induced oxidative stress in chronic obstructive pulmonary disease994-1001Am J Respir Crit Care Med17286Breath Tests Case-Control Studies Comet Assay DNA Damage/physiology Exercise Test Exercise Therapy/*adverse effects/methods Exercise Tolerance/*physiology Female Forced Expiratory Volume Humans Hydrogen Peroxide/analysis Leukocytes, Mononuclear/metabolism Male Malondialdehyde/blood/urine Middle Aged Oxidative Stress/*physiology Pulmonary Disease, Chronic Obstructive/*metabolism/physiopathology/*rehabilitation Reactive Oxygen Species/metabolism Research Support, Non-U.S. Gov't Rest/physiology Severity of Illness Index Time Factors Uric Acid/blood Vital CapacityOct 15OThe effect of exercise at different intensities as well as the effect of intensive supervised pulmonary rehabilitation on oxidative stress were studied for chronic obstructive pulmonary disease (COPD). Eleven patients with COPD and 11 healthy age-matched control subjects performed a maximal and submaximal exercise cycle ergometry test at 60% of peak workload. Patients with COPD performed these tests before and after 8 wk of pulmonary rehabilitation. Measurements were done before, immediately after, and 4 h after both exercise tests. At rest, increased oxidative stress was observed in patients compared with control subjects, as measured by urinary malondialdehyde (MDA; p < 0.05) and hydrogen peroxide (H2O2) in breath condensate (p < 0.05). In healthy control subjects, a significant increase in urinary MDA was observed 4 h after both exercise tests (p = 0.05), whereas H2O2 significantly increased immediately after maximal exercise (p < 0.05). In patients with COPD, before rehabilitation, reactive oxygen species-induced DNA damage in peripheral blood mononuclear cells, urinary MDA, and plasma uric acid were significantly increased after both exercise tests (p < 0.05), whereas no significant increase was observed in plasma MDA. In contrast, exhaled H2O2 was only significantly increased after maximal exercise (p < 0.02). Although after rehabilitation peak workload was increased by 24%, a similar oxidative stress response was found. Remarkably, a decrease in reactive oxygen species-induced DNA damage was detected after exercise at submaximal intensity despite increased exercise duration of 73%. In summary, patients with COPD had increased pulmonary and systemic oxidative stress both at rest and induced by exercise. In addition, pulmonary rehabilitation increased exercise capacity and was associated with reduced exercise-induced oxidative stress.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16040783 !1073-449X (Print) Journal Article16040783sDepartment of Respiratory Medicine, University of Maastricht, Maastricht, The Netherlands. e.mercken@pul.unimaas.nl ~?4*Qiao, H. Wang, G. F. Zhang, H. Ding, C. M.2005q[The interleukin-6 level in exhaled breath condensate of patients with obstructive sleep apnea-hypopnea syndrome]364-7Zhonghua Jie He He Hu Xi Za Zhi286JunOBJECTIVE: To explore whether the airway inflammation marker in exhaled breath condensate is increased in obstructive sleep apnea-hypopnea syndrome (OSAHS). METHODS: Thirty-one patients with OSAHS (15 smokers and 16 non-smokers) and 10 healthy age-matched and weight-matched controls were included in the study. Exhaled breath condensate (EBC) was collected before and after sleep at the same night from both groups. Interleukin-6 (IL-6) in EBC was measured by a specific enzyme immunoassay. RESULTS: (1) There was no difference in the pre-sleep IL-6 level among OSAHS smoker group, OSAHS non-smoker group and the control subjects (F = 0.515, P > 0.05). Compared with the level of pre-sleep [(2.5 +/- 1.0) ng/L in OSAHS smoker, and (2.3 +/- 0.8) ng/L in OSAHS non-smoker], the post-sleep level of IL-6 was elevated significantly in EBC from OSAHS patients of non-smokers [(3.1 +/- 1.2) ng/L] and smokers [(3.7 +/- 1.9) ng/L, P < 0.05]. Nevertheless, IL-6 level from the control group showed a reverse change. IL-6 was decreased significantly [(2.0 +/- 0.8) ng/L in pre-sleep vs (2.7 +/- 1.0) ng/L in post-sleep] after sleep in this group. There was no difference in post-sleep IL-6 level between OSAHS smokers [(3.7 +/- 1.9) ng/L] and non-smokers [(3.1 +/- 1.2) ng/L, P > 0.05]. A significant higher IL-6 level was observed in both OSAHS smokers [(3.7 +/- 1.9) ng/L] and non-smokers [(3.1 +/- 1.2) ng/L] compared with the controlled group [(2.0 +/- 0.8) ng/L, P < 0.05]. IL-6 level in EBC was correlated positively with AHI (r = 0.441, P < 0.05), ODI(4) (r = 0.533, P < 0.05), and negatively with minimal oxygen saturation (r = -0.529, P < 0.05). CONCLUSIONS: These findings suggest that inflammation was characteristic in the airways of OSAHS patients. Nocturnal hypoxia could be responsible for this change. The levels of IL-6 in EBC are associated with the severity of OSAHS and may prove to be useful in monitoring of airway inflammation in OSAHS.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16008970 !1001-0939 (Print) Journal Article16008970\Department of Respiratory Medicine, Peking University First Hospital, Beijing 100034, China. ~?54Larstad, M. Soderling, A. S. Caidahl, K. Olin, A. C.2005Selective quantification of free 3-nitrotyrosine in exhaled breath condensate in asthma using gas chromatography/tandem mass spectrometry134-44 Nitric Oxide132>Adult Aged Asthma Biological Markers/analysis Case-Control Studies Comparative Study *Exhalation Female Humans Inflammation/diagnosis Male *Mass Fragmentography Middle Aged Oxidative Stress Reproducibility of Results Research Support, Non-U.S. Gov't Sensitivity and Specificity Tyrosine/*analogs & derivatives/analysisSepReactive nitrogen species can cause oxidative modifications of certain amino acid residues in proteins, notably the modification of tyrosine to 3-nitrotyrosine (3-NT), which is a potentially useful marker of oxidative stress. Since lung diseases are associated with airway inflammation and oxidative stress, quantification of 3-NT in exhaled breath condensate (EBC) may provide a non-invasive means for monitoring ongoing inflammatory processes. 3-NT-like immunoreactivity has previously been detected in EBC, but no definitive evidence for the presence of 3-NT in EBC is available. Here, a method based on gas chromatography/negative ion chemical ionization/tandem mass spectrometry was established for the quantification of free 3-NT in EBC. The detection limit was 0.56 pM (corresponding to 3.0 amol microl(-1) sample injected) and the method was found to give linear results (r2 > 0.999) in the concentration range of 0-5.0 nM. The coefficient of variation (CV) for within-day and between-day precision were 11 and 12%, respectively. No artifactual nitration was observed during sample processing. The method was applied to study subjects with asthma (n = 8), and healthy subjects (n = 10), but only a slight non-significant increase in 3-NT levels was found in the former group (median [interquartile ranges]; 99 [50-547] amol s(-1) vs. 75 [35-147] amol s(-1)). No correlation with exhaled nitric oxide (NO), pulmonary function or EBC levels of total protein was observed. The 3-NT levels were much lower compared to previously reported levels, based on immunochemical measurements. The method does not allow the simultaneous quantification of tyrosine in samples.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=16006156 !1089-8603 (Print) Journal Article16006156}Department of Occupational Medicine, The Sahlgrenska Academy at Goteborg University, Goteborg, Sweden. mona.larstad@amm.gu.se ~?6 Rahman, I.2005RThe role of oxidative stress in the pathogenesis of COPD: implications for therapy175-200Treat Respir Med43Antioxidants/*therapeutic use Humans *Oxidative Stress Practice Guidelines Pulmonary Disease, Chronic Obstructive/drug therapy/*etiology Reactive Oxygen Species/metabolism Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.Chronic inflammation and oxidative stress are important features in the pathogenesis of COPD. The increased oxidative stress in patients with COPD is the result of an increased burden of inhaled oxidants, as well as increased amounts of reactive oxygen species (ROS) generated by various inflammatory, immune and epithelial cells of the airways. Oxidative stress has important implications on several events of lung physiology and for the pathogenesis of COPD. These include oxidative inactivation of antiproteases and surfactants, mucus hypersecretion, membrane lipid peroxidation, mitochondrial respiration, alveolar epithelial injury, remodeling of extracellular matrix, and apoptosis. An increased level of ROS produced in the airways is reflected by increased markers of oxidative stress in the airspaces, sputum, breath, lungs, and blood in patients with COPD. The biomarkers of oxidative stress such as H2O2, F2-isoprostanes, malondialdehyde and 4-hydroxy-2-nonenal have been successfully measured in breath condensate. ROS and aldehydes play a key role in enhancing the inflammation through the activation of mitogen-activated protein kinases and redox-sensitive transcription factors such as nuclear factor kappa B and activator protein-1. Oxidative stress also alters nuclear histone acetylation and deacetylation leading to increased gene expression of pro-inflammatory mediators in the lung. Oxidative stress may play a role in the poor clinical efficacy of corticosteroids in the treatment of COPD. Since a variety of oxidants, free radicals, and aldehydes are implicated in the pathogenesis of COPD it is likely that a combination of antioxidants may be effective in the treatment of COPD. Antioxidant compounds may also be of therapeutic value in monitoring oxidative biomarkers indicating disease progression. Various approaches to enhance the lung antioxidant screen and the clinical effectiveness of antioxidant compounds in the treatment of COPD are discussed.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15987234 (1176-3450 (Print) Journal Article Review15987234Department of Environmental Medicine, Division of Lung Biology and Disease, University of Rochester Medical Center, Rochester, New York 14642, USA. irfan_rahman@urmc.rochester.edu ~?7HMascia, K. Borish, L. Patrie, J. Hunt, J. Phillips, C. D. Steinke, J. W.2005eChronic hyperplastic eosinophilic sinusitis as a predictor of aspirin-exacerbated respiratory disease652-7Ann Allergy Asthma Immunol946Adult Aspirin/*adverse effects Asthma/diagnosis/immunology Case-Control Studies Chronic Disease Cohort Studies Comparative Study Drug Hypersensitivity/complications/*diagnosis Eosinophilia/*diagnosis/immunology Female Humans Hyperplasia Male Middle Aged Paranasal Sinuses/pathology/radiography Prognosis Research Support, N.I.H., Extramural Research Support, U.S. Gov't, P.H.S. Respiratory Tract Diseases/*diagnosis Sinusitis/immunology/*radiography Tomography, X-Ray ComputedJunBACKGROUND: Aspirin-exacerbated respiratory disease (AERD) is a disease of intense eosinophilic inflammation that can produce fibrosis, hyperplasia, and remodeling. OBJECTIVE: To investigate the usefulness of quantifying severity of chronic hyperplastic eosinophilic sinusitis in predicting the presence of AERD. METHODS: Data were compared between asthmatic patients who reported exacerbations after aspirin ingestion and those who did not. The primary outcome measure was severity of sinusitis using a validated computed tomography (CT) scan-based scoring system. Indices of lower airway remodeling and other markers of inflammation were also evaluated. RESULTS: Twenty-one patients with AERD were compared with 19 patients with aspirin-tolerant asthma (ATA). Patients were well matched for asthma severity as shown by their similar lung function as measured by postbronchodilator forced expiratory volume in 1 second. Patients with AERD were distinguished by their sinus CT scores (AERD patients: 16.9; 95% confidence interval [CI], 13.4-21.3; ATA patients: 6.2; 95% CI, 4.2-9.1; P < .001), and they were considerably more likely to have nasal polyps (AERD patients: 90%; ATA patients: 26%; P < .001). In addition, AERD patients demonstrated increased total lung capacity (AERD patients: 107.9%; 95% CI, 99.9%-117.6%; ATA patients: 98.0%; 95% CI, 93.7%-102.5%; P = .05), reflecting a trend toward increased air trapping. No significant differences occurred in diffusing capacity, exhaled nitric oxide, eosinophilia, or exhaled breath condensate pH. CONCLUSIONS: AERD can be distinguished from ATA by the extent of hyperplasia on CT scan and the presence of nasal polyps. We hypothesize that AERD represents a remodeling process that affects both the upper and lower airways.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15984597 !1081-1206 (Print) Journal Article15984597iDepartment of Medicine, University of Virginia Health Systems, Charlottesville, Virginia 22908-1355, USA.~?8eWhite, D. C. Geyer, R. Cantu, J. Jo, S. C. Peacock, A. D. Saxton, A. M. Mani, S. Jett, M. Moss, O. R.2005Feasibility of assessment of regulatory lipids in breath condensate as potential presymptomatic harbingers of pulmonary pathobiology293-302J Microbiol Methods623Actinobacillus pleuropneumoniae/pathogenicity Animals Breath Tests/*methods Cell Line Chromatography, High Pressure Liquid Eicosanoids/analysis Enterotoxins/toxicity Humans Lipids/*analysis Lipopolysaccharides/toxicity Lung Diseases/*diagnosis/metabolism Mass Fragmentography Phospholipids/analysis/chemistry Platelet Activating Factor/analogs & derivatives/analysis Rats Research Support, Non-U.S. Gov't Respiratory Tract Infections/diagnosis/metabolism Spectrometry, Mass, Electrospray Ionization Sus scrofaSep Regulatory lipids from the airway surface readily form aerosols that can be recovered non-invasively by cooling expired breath to form breath condensate (BC). Regulatory lipids have been detected previously utilizing enzyme-linked-immunosorbent serologic assay (ELISA). Here we test the feasibility of assessment of regulatory lipids in BC by mass spectrometry so presently unknown lipid regulatory components can be detected without addition of specific antibodies as in the ELISA procedure. Baseline regulatory lipids were detected in >pg/mL BC in control animals or human lung tissue culture cells. In nearly every case animals exposed to toxins or infectious bacteria showed increases in the BC regulatory components. Lipids were recovered from BC by solid phase extraction. Phosphatidylcholine (PC) based lipids were detected as the progenitor (parent) ions of isomers that fragmented in producing product positive ions at m/z 184 (of phosphocholine) in tandem MS using capillary HPLC and electrospray ionization. BC eicosanoids such as prostaglandins, thromboxane, and isoprostanes require capillary gas chromatography for separation and detection that necessitates methoximation, pentafluorobenzyl (PFB) ester formation, and trimethyl silylation of hydroxyls prior to gas chromatography/ion trap tandem mass spectrometry of negative ions after chemical ionization (NICI). Tetradeuterated internal standards were utilized for quantitation with the GC/NICI/MS. Changes in concentrations of lipids and eicosanoids were observed in piglets, and rats exposed to aerosolized 100 mug/kg lipopolysaccharide (LPS), or 50 mug/kg and 150 mug/kg aerosolized Staphylococcal enterotoxin B (SEB) in BC as well as in human THP-1 cell culture cell supernatants and bronchoalveolar lavage (BAL) samples in rats. Responses of the molecular species of phosphatidylcholines (PCs), platelet activating factors (PAFs) and specific eicosanoids correlated to the toxin and bacterial infections suggesting that patterns of differential responses could be detected with further experimentation. Initial targets included prostaglandins (PGE(2), PGF(2alpha)), thromboxane (TXB2), and prostacyclin (as 6-Keto PGF(1alpha)) that show differential responses to inflammation, the leukotriene (LTB4) and PGD2 for allergic responses, isoprostanes (8-iso-PGF(2alpha)) for free radical oxidative stress responses, and HETEs for differential lipoxygenase activities. PAFs and lysoPAFs have been shown to increase with inflammation and in the feasibility experiments reported here. Preliminary studies show pulmonary responses of piglets to intrathecal exposure of toxicants (LPS and SEB) or infections with Actinobacillus pleuropneumoniae induce increased levels of lipids and two eicosanoids with the suggestion that differential patterns might be detected with expanded testing. Preliminary experience indicates numerous other eicosanoids were available for assay in BC. This suggests an important potential application of BC to observe a wide array of factors to establish comprehensive profiles for physiological and pathophysiological states. Ultimately this technique could be used as a non-invasive possibly presymptomatic assessment of pulmonary pathobiology.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15963585 !0167-7012 (Print) Journal Article15963585bCenter for Biomarker Analysis, University of Tennessee, Knoxville, TN, USA. mailto:Dwhite1@utk.edu C~?9Carpagnano, G. E. Foschino-Barbaro, M. P. Mule, G. Resta, O. Tommasi, S. Mangia, A. Carpagnano, F. Stea, G. Susca, A. Di Gioia, G. De Lena, M. Paradiso, A.2005h3p microsatellite alterations in exhaled breath condensate from patients with non-small cell lung cancer738-44Am J Respir Crit Care Med1726_Aged Carcinoma, Non-Small-Cell Lung/*genetics/pathology Comparative Study DNA, Neoplasm/blood/genetics Exhalation/*genetics Female Humans Loss of Heterozygosity Lung Neoplasms/*genetics/pathology Male *Microsatellite Repeats Middle Aged Neoplasm Staging Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Smoking/adverse effectsSep 15The still-high mortality for lung cancer urgently requires the availability of new, noninvasive diagnostic tools for use in early diagnosis and screening programs. Recently, exhaled breath condensate (EBC) has been proposed as a useful tool to obtain biological information on lung cancer disease. This study provides, for the first time, evidence that DNA alterations already described in lung cancer are detectable in EBC from patients with non-small cell lung cancer (NSCLC) and in healthy subjects. Thirty patients with histologic evidence of NSCLC and 20 healthy subjects were enrolled in the present study. All subjects had allelotyping analysis of DNA from EBC (EBC-DNA) and from whole blood (WB-DNA) of a selected panel of five microsatellites (D3S2338, D3S1266, D3S1300, D3S1304, D3S1289) located in chromosomal region 3p. Results from healthy subjects and subjects with cancer, and from EBC and WB, were compared. In addition, the relationships with smoking habit and clinicopathologic tumor features were considered. Microsatellite alterations (MAs) were found in 53% of EBC-DNA and in 10% of WB-DNA loci investigated in patients with NSCLC (p < 10(-6)); conversely, MAs were present only in 13% of EBC-DNA and in 2% of WB-DNA informative loci in healthy subjects. In patients with NSCLC, a direct association between number of MAs detected in EBC-DNA and tobacco consumption was observed. We conclude that EBC-DNA is highly sensitive in detecting MA information unique to patients with lung cancer. Furthermore, MA information seems to be directly related with tobacco consumption, and is potentially applicable to screening and early diagnostic programs for patients with NSCLC.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15947287 !1073-449X (Print) Journal Article15947287UInstitute of Respiratory Disease, University of Foggia, Italy. ge.carpagnano@unifg.it~?:Dobrykh, V. A. Mun, I. E.2005N[Some problems in the study of expired air condensate in respiratory diseases]33-5Probl Tuberk Bolezn Legk3Adult Breath Tests Comparative Study Dust/*analysis Female Gases/*chemistry Humans Male Reference Values Respiratory Tract Diseases/*diagnosis/metabolism Water/*analysisfhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15945450 !1728-2993 (Print) Journal Article15945450 ~?; Montuschi, P.20058Exhaled breath condensate analysis in patients with COPD22-34Clin Chim Acta3561-2Breath Tests/*methods Dinoprost/analogs & derivatives/analysis Dinoprostone/analysis Humans Hydrogen Peroxide/analysis Hydrogen-Ion Concentration Leukotriene B4/analysis Malondialdehyde/analysis Pulmonary Disease, Chronic Obstructive/diagnosis/*metabolism Smoking/metabolismJunExhaled 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.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15936301 (0009-8981 (Print) Journal Article Review15936301Department of Pharmacology, Faculty of Medicine, Catholic University of the Sacred Heart, Largo F. Vito, 1, 00168 Rome, Italy. pmontuschi@rm.unicatt.it ~?<[Effros, R. M. Peterson, B. Casaburi, R. Su, J. Dunning, M. Torday, J. Biller, J. Shaker, R.2005nEpithelial lining fluid solute concentrations in chronic obstructive lung disease patients and normal subjects1286-92J Appl Physiol994Aged Body Fluids/*metabolism Body Water/metabolism Calcium/blood/metabolism Case-Control Studies Epithelium/metabolism Exhalation Female Humans Isotonic Solutions/metabolism Lung/*metabolism Magnesium/blood/metabolism Male Middle Aged Osmolar Concentration Potassium/blood/metabolism Pulmonary Disease, Chronic Obstructive/*metabolism Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Saliva/metabolism Sodium/blood/metabolismOctThe exhaled breath condensate (EBC) method represents a new, noninvasive way to detect inflammatory and metabolic markers in the fluid that covers the airways [epithelial lining fluid (ELF)]. However, respiratory droplets represent only a very small and variable fraction of the EBC, most (approximately 99.99%) of which is water vapor. Our objective was to show that ELF concentrations could be calculated from EBC values by using any of three dilutional indicators (urea, total cations, and conductivity) in nine normal and nine chronic obstructive lung disease (COPD) subjects. EBC concentrations of Na(+), K(+), Ca(2+), Mg(2+), total cations, urea, and conductivity varied over a 10-fold range among individuals, but concentrations of these constituents (except Ca(2+)) remained well correlated (r(2) = 0.44-0.83, P < 0.001). Dilution (D) of respiratory droplets in water vapor was calculated by dividing plasma concentrations of the dilutional indicators by EBC concentrations. Estimates of D were not significantly different among these indicators, and urea D averaged 10,800 +/- 2,100 (SE) in normal and 12,600 +/- 3,300 in COPD subjects. Although calculated Na(+) concentrations in the ELF were less than one-half those in plasma, and concentrations of K(+), Ca(2+), and Mg(2+) exceeded those in plasma, total cation concentrations in ELF were not significantly different from those in plasma, indicating that ELF is isotonic in both normal and COPD subjects. EBC amylase concentrations (measured with an ultrasensitive procedure) indicated that saliva represented <10% of the respiratory (ELF) droplets in all but three samples. Dilutional and salivary markers are essential for interpretation of EBC studies.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15920100 !8750-7587 (Print) Journal Article15920100Los Angeles Biomedical Research Institute at Harbor-University of California-Los Angeles Medical Center, Torrance, 90505, USA. reffros@labiomed.org~?=Tufvesson, E. Bjermer, L.2006`Methodological improvements for measuring eicosanoids and cytokines in exhaled breath condensate34-8 Respir Med1001Jan:BACKGROUND: Exhaled breath condensate (EBC) is simple to collect and as such a non-invasive method that has attracted substantial interest in the last few years. However, several methodological concerns have been raised and it has been difficult to reproduce results between different centres. Because of low concentrations of inflammatory markers, potential loss in the sampling system may have great influence. The aim of the present study was to se if evaporation and plastic coating could facilitate detection. METHODOLOGY: Through methodological improvements, we have now made it possible to measure EBC concentrations of eicosanoids and cytokines in our system. Due to absorbance of both fatty acid derivates and proteins to several plastics, the first step is coating of all surfaces with bovine serum albumin and Tween 20. Since several assays are sensitive to these factors, the methodology has to be standardised to avoid false results. Secondly, larger amounts of EBC have to be vacuum-dried, and thereafter resolved in the respective assay buffers. The EBCs have to be concentrated 5-10 times, depending on samples and assay sensitivity. RESULTS: Due to these improvements we can measure, for example, cysteinyl-leukotrienes, leukotriene B4, prostaglandin E and 8-isoprostane. High sensitivity assays have also made it possible to measure cytokines, for example, interleukin (IL)-1beta, IL-8 and IL-13. SUMMARY: We are aware of different results from other labs. However, it seems essential to coat and to concentrate the samples in order to achieve reliable and measurable results.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15894480 !0954-6111 (Print) Journal Article15894480wDepartment of Respiratory Medicine and Allergology, University Hospital, 221 85 Lund, Sweden. ellen.tufvesson@med.lu.se ~?>QKostikas, K. Gaga, M. Papatheodorou, G. Karamanis, T. Orphanidou, D. Loukides, S.2005cLeukotriene B4 in exhaled breath condensate and sputum supernatant in patients with COPD and asthma1553-9Chest1275Adult Aged Asthma/*diagnosis Breath Tests Cross-Sectional Studies Diagnosis, Differential Humans Leukotriene B4/*analysis Male Middle Aged Prospective Studies Pulmonary Disease, Chronic Obstructive/*diagnosis Sputum/*chemistryMay3 STUDY OBJECTIVES: Some patients with COPD present with significant reversibility of airflow limitation after receiving bronchodilation therapy. Leukotriene B(4) (LTB(4)) has been implicated in the pathophysiology of both COPD and asthma. We tested the hypothesis that COPD patients with airflow reversibility and asthmatic patients who smoke might have similar levels of LTB(4) in exhaled breath condensate (EBC) and sputum supernatant. The repeatability and stability of LTB(4) measurements were additionally studied. DESIGN: Prospective, cross-sectional study. PATIENTS OR PARTICIPANTS: We studied 30 patients with COPD (15 smokers [FEV(1), 56% predicted; SD, 6% predicted]; 15 patients with significant reversibility in airway obstruction after bronchodilation [FEV(1), 14% predicted; SD, 2% predicted]). Fifteen asthmatic patients who smoked, with similar FEV(1) and reversibility were also studied. Ten healthy smokers served as control subjects. SETTING: A hospital research laboratory. INTERVENTIONS: Spirometry and reversibility testing were performed on the first visit. On the following day, EBC was collected for the measurement of LTB(4), and induced sputum was collected for differential cell counts and LTB(4) measurement in the sputum supernatant. MEASUREMENTS AND RESULTS: LTB(4) levels in EBC [mean (SD)] were increased in COPD patients (mean, 86.7 pg/mL; SD, 19 pg/mL) and asthmatic patients (mean, 97.5 pg/mL; SD, 15 pg/mL) compared to control subjects (mean, 32.3 pg/mL; SD, 10 pg/mL; p < 0.0001 for both groups). COPD patients with airflow reversibility (mean, 99.8 pg/mL; SD, 12 pg/mL) had values similar to those of asthmatic patients (mean, 97.5 pg/mL; SD, 15 pg/mL; p = 0.2) and higher than those of COPD patients without airflow reversibility (mean, 73.7 pg/mL; SD, 17 pg/mL; p = 0.002). Similar results were observed in the sputum supernatant. Measurements of LTB(4) in EBC and sputum were repeatable on two consecutive days, but measurements in the frozen samples of EBC and sputum were not stable after 3 weeks. CONCLUSIONS: Patients with asthma and reversible COPD presented with higher LTB(4) values compared to patients with nonreversible COPD and healthy smokers. This difference may be mainly attributed to the presence of reversibility in airway obstruction, probably as part of a common underlying inflammatory process.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15888827 !0012-3692 (Print) Journal Article15888827UPneumonology Department, University of Athens, Athens Chest Hospital, Athens, Greece.~??NFranklin, P. Moeller, A. Hall, G. L. Horak, F., Jr. Patterson, H. Stick, S. M.2006DVariability of nitric oxide metabolites in exhaled breath condensate123-9 Respir Med1001JanThe collection of exhaled breath condensate (EBC) is simple and non-invasive, however, there are few data on the methodological aspects affecting concentrations of compounds in EBC. The aim of this study was to investigate methodological issues for measuring nitric oxide metabolites (NO(x)) in EBC. Twenty-five healthy adults (12 females, age range 23-55 years) and 22 children (11 females, age range 7-6 years) were recruited for studies investigating inter- and intra-day repeatability, repeatability with controlled expiratory flows and temperature, flow dependence, and analytical variability of EBC NO(x). Both intra- and inter-day repeatability was poor with a coefficient of repeatability of 103.4% of the mean difference between intra-day (15 min) measures and 118.6% of inter-day (24 h) differences. Repeatability was not improved when expiratory flow and temperature of the collection device were controlled. However, some of the variability (approximately 50%) may be accounted for by variability in the analytical technique (analytical variability) and this may result from difficulties in controlling for contamination. NO(x) levels were not affected by different expiratory flows in either adults or children but there was still significant variation within individuals. Levels of NO(x) in EBC seem to be highly variable and this needs to be considered if EBC NO(x) is to be used in clinical studies.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15885998 !0954-6111 (Print) Journal Article15885998School of Paediatrics and Child Health, University of Western Australia, Roberts Road, Perth, Western Australia 6008, Australia. peterf@ichr.uwa.edu.aui~?@UCsoma, Z. Huszar, E. Vizi, E. Vass, G. Szabo, Z. Herjavecz, I. Kollai, M. Horvath, I.2005WAdenosine level in exhaled breath increases during exercise-induced bronchoconstriction873-8 Eur Respir J255Adenosine/*blood Adult Asthma, Exercise-Induced/*blood/chemically induced/diagnosis *Breath Tests Comparative Study Exercise/physiology Female Humans Male Methacholine Chloride Research Support, Non-U.S. Gov't Respiratory Function Tests RestMay<In asthmatic patients, airway obstruction provoked by exercise challenge is accompanied by an increase in plasma adenosine level. In this study, the current authors investigated if exercise-induced bronchoconstriction was associated with local changes of adenosine concentration in the airways. Oral exhaled breath condensate (EBC) collection (5-min duration) and forced expiratory volume in one second (FEV1) measurements were performed at rest (baseline) and 4-8 times after treadmill exercise challenge in healthy and asthmatic subjects. Adenosine concentration in EBC was determined by HPLC. Observations indicated that physical exercise results in bronchoconstriction together with a significant increase of adenosine level in EBC in asthmatic patients (mean+/-sd maximal fall in FEV1 27+/-13%; associated increase in adenosine 110+/-76% as compared to baseline), but not in healthy control subjects. Exercise-induced changes in adenosine concentration correlated significantly with the fall in FEV1 values in asthmatic patients. In conclusion, the observed increase in adenosine concentration of oral exhaled breath condensate most probably reflects changes in the airways during exercise-induced bronchoconstriction. Due to its known bronchoconstrictor property in asthma, adenosine may contribute to the development of bronchospasm.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15863645 J0903-1936 (Print) Clinical Trial Controlled Clinical Trial Journal Article15863645National Koranyi Institute for Tuberculosis and Pulmonology, Dept of Pathophysiology, Budapest, Piheno u.1. P.O. Box 1, H-1529, Hungary.c~?A Fudge, B. M.2005:Design parameters of a low flow rate sidestream capnograph153-8Biomed Sci Instrum41Capnography/*instrumentation/*methods Carbon Dioxide/*analysis *Computer-Aided Design Equipment Design Equipment Failure Analysis Microfluidics/*instrumentation/*methodsThe use of sidestream capnometers, with a sampling rate of 150-250 cc/min, as a means of measuring a patient's expired CO2 (ETCO2) and respiratory rate, has been a common practice for many years. However, in recent years, there has been a focus on lower flow rate sampling sidestream systems due to the benefits of less loss of tidal volume for patients, such as infants or neonates. When developing a sidestream system, four principle issues must be considered; 1) The signal fidelity of the gas sample must be sufficiently maintained from the sampling site to the measurement site. 2) Condensate from a patient's breath, as well as blood, mucus, or other contaminates often pose problems for sidestream systems and requires mitigation. 3) The mechanics of transporting a gas sample at a constant flow rate through the sampling system, regardless of atmospheric or clinical conditions must be developed. 4) The physics of handling CO2 gas throughout the transport process must be understood in order to ensure accurate readings. These issues lead to a complex web of interrelations that are explored in the development of a low flow rate sidestream capnometer.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15850097 40067-8856 (Print) Evaluation Studies Journal Article15850097 ~?BSKirschvink, N. Marlin, D. Delvaux, F. Leemans, J. Clercx, C. Sparkes, A. Gustin, P.2005Collection of exhaled breath condensate and analysis of hydrogen peroxide as a potential marker of lower airway inflammation in cats385-96Vet J1693UAirway Obstruction/diagnosis/*veterinary Animals Biological Markers/analysis/metabolism Breath Tests/*methods Bronchoalveolar Lavage Fluid/cytology Bronchoscopy/veterinary Case-Control Studies Cat Diseases/*diagnosis/pathology Cats Female Hydrogen Peroxide/*analysis/metabolism Male Predictive Value of Tests Research Support, Non-U.S. Gov'tMay)The objective of this study was to describe a standardised and non-invasive method for exhaled breath condensate (EBC) collection in cats and to test whether determination of hydrogen peroxide (H(2)O(2)) in EBC might be used as marker of lower airway inflammation. The technique of barometric whole body plethysmography for cats was combined with a system to condense the effluent air from the plethysmograph, allowing simultaneous EBC collection and respiratory pattern measurement. H(2)O(2) was determined spectrophotometrically. Eighteen experimental cats were used to investigate the impact on EBC volume and EBC H(2)O(2) of plethysmograph ventilation rate, collection duration, sample stability, within-day and day-to-day variability. After determination of a standardised EBC collection procedure, correlation analyses between EBC H(2)O(2) and bronchoalveolar lavage (BAL) cytology of healthy and allergen-challenged Ascaris suum (AS)-sensitised cats were performed. A significant and positive correlation between EBC H(2)O(2) and bronchoalveolar lavage (BAL) neutrophil% was found in healthy cats (P < 0.001, r = 0.55), whereas in AS-sensitised cats, correlation with BAL eosinophil% was significant (P < 0.005, r = 0.61). H(2)O(2) was increased after an allergen challenge in AS-sensitised cats (n = 6, 0.56+/-0.12 versus 1.08+/-0.35 micromol/L, P < 0.05). This study proposes a non-invasive, well tolerated and repeatable method of EBC collection for cats and suggests that EBC H(2)O(2) might be used as non-invasive biomarker for monitoring lower airway inflammation.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15848781 41090-0233 (Print) Evaluation Studies Journal Article15848781Section of Pharmacology, Pharmacotherapy and Toxicology, Department of Functional Sciences B41, Faculty of Veterinary Medicine, University of Liege, 4000 Liege, Belgium. Nathalie.Kirschvink@ulg.ac.be ~?CKLeung, T. F. Wong, G. W. Ko, F. W. Li, C. Y. Yung, E. Lam, C. W. Fok, T. F.2005jAnalysis of growth factors and inflammatory cytokines in exhaled breath condensate from asthmatic children66-72Int Arch Allergy Immunol1371Adolescent Asthma/*immunology/*metabolism Breath Tests Child Cytokines/analysis/*metabolism Epidermal Growth Factor/analysis Female Forced Expiratory Volume Growth Substances/analysis/*metabolism Humans Immunoglobulin E/blood Interleukin-4/analysis Male Nitric Oxide/analysis Platelet-Derived Growth Factor/analysis Research Support, Non-U.S. Gov't Statistics, Nonparametric Tumor Necrosis Factor-alpha/analysis Vascular Endothelial Growth Factor A/analysis Vital CapacityMayBACKGROUND: Vascular endothelial growth factor (VEGF), AA isoform of platelet-derived growth factor (PDGF-AA), and epidermal growth factor (EGF) are involved in the pathogenesis of airway inflammation in asthma. These molecules are closely associated with cytokines such as tumor necrosis factor-alpha (TNF-alpha) and interleukin (IL)-4. This study investigates the relation between childhood asthma and levels of these mediators in exhaled breath condensate (EBC). METHODS: EBC was collected from asthmatic children and controls using a disposable collection kit, and the concentrations of VEGF, PDGF-AA, EGF, TNF-alpha and IL-4 in EBC were measured using sandwich enzyme immunoassays. Exhaled nitric oxide concentration was measured by a chemiluminescence analyzer. RESULTS: Thirty-five asthmatic patients aged between 7 and 18 years and 11 controls were recruited. Sixteen patients had intermittent asthma (IA) whereas 19 of them suffered from persistent asthma (PA). A significant correlation was found between IL-4 and TNF-alpha in EBC (rho = 0.374, p = 0.010). PDGF-AA levels in EBC were higher in subjects with diminished FEV1 (p = 0.023) whereas IL-4 concentrations were increased in asthmatics (p = 0.007) as well as subjects with increased plasma total IgE (p = 0.033). Patients with PA receiving high-dose inhaled corticosteroid (ICS) had higher EBC IL-4 concentration than those on low-dose ICS (p = 0.007). Linear regression revealed that PDGF-AA levels in EBC were negatively associated with FEV1 percentage (beta = -0.459, p = 0.006) among the asthmatic patients. CONCLUSIONS: IL-4 in EBC is increased in childhood asthma, and growth factors are detectable in a significant proportion of these children. Increased PDGF-AA is found in asthmatics with more severe airflow limitation.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15832052 !1018-2438 (Print) Journal Article15832052kDepartment of Paediatrics, The Chinese University of Hong Kong, Hong Kong SAR, China. leung2142@cuhk.edu.hk $~?DHolz, O. Tal-Singer, R. Kanniess, F. Simpson, K. J. Gibson, A. Vessey, R. S. Janicki, S. Magnussen, H. Jorres, R. A. Richter, K.2005rValidation of the human ozone challenge model as a tool for assessing anti-inflammatory drugs in early development498-503J Clin Pharmacol455Administration, Inhalation Administration, Oral Adult Androstadienes/administration & dosage/pharmacology Anti-Inflammatory Agents/administration & dosage/pharmacology Bronchial Provocation Tests/*methods Cross-Over Studies Double-Blind Method Drug Evaluation/*methods Female Humans Male Models, Biological Ozone/*diagnostic use Prednisolone/administration & dosage/pharmacology Research Support, Non-U.S. Gov't Respiratory Function Tests Sputum/cytology/metabolismMayThis study aimed to test the utility of the ozone challenge model for profiling novel compounds designed to reduce airway inflammation. The authors used a randomized, double-dummy, double-blind, placebo-controlled 3-period crossover design alternating single orally inhaled doses of fluticasone propionate (inhaled corticosteroids, 2 mg), oral prednisolone (oral corticosteroids, 50 mg), or matched placebo. At a 2-week interval, 18 healthy ozone responders (>10% increase in sputum neutrophils) underwent a 3-hour ozone (250 ppb)/intermittent exercise challenge starting 1 hour after drug treatment. Airway inflammation was assessed at 2 hours (breath condensate) and 3 hours (induced sputum) after ozone challenge. Compared to placebo, pretreatment with inhaled corticosteroids or oral corticosteroids resulted in a significant reduction (mean [95% confidence interval]) of sputum neutrophils by 62% (35%, 77%) and 64% (39%, 79%) and of sputum supernatant myeloperoxidase by 55% (41%, 66%) and 42% (25%, 56%), respectively. The authors conclude that an optimized ozone challenge model (including ozone responders and ensuring adequate drug levels during exposure) may be useful for testing novel anti-inflammatory compounds in early development.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15831772 L0091-2700 (Print) Clinical Trial Journal Article Randomized Controlled Trial15831772rHospital Grosshansdorf, Center for Pneumology and Thoracic Surgery, Wohrendamm 80, D-22927 Grosshansdorf, Germany. ~?E[Carraro, S. Corradi, M. Zanconato, S. Alinovi, R. Pasquale, M. F. Zacchello, F. Baraldi, E.2005tExhaled breath condensate cysteinyl leukotrienes are increased in children with exercise-induced bronchoconstriction764-70J Allergy Clin Immunol1154Acetates/therapeutic use Adolescent Ammonia/analysis/metabolism Asthma, Exercise-Induced/drug therapy/*physiopathology Biological Markers/*analysis Breath Tests Bronchoconstriction/drug effects Child Cysteine/*analysis/drug effects/metabolism Exhalation Forced Expiratory Volume Humans Leukotriene Antagonists/therapeutic use Leukotriene B4/analysis/metabolism Leukotrienes/*analysis/metabolism Nitric Oxide/analysis/metabolism Quinolines/therapeutic use Respiratory Function TestsApr'BACKGROUND: It is recognized that airway inflammation has a central role in the pathogenesis of asthma, but how it relates to exercise-induced bronchoconstriction (EIB) is not completely understood. OBJECTIVE: The aim of our study was to investigate the relationship between EIB and baseline concentrations of cysteinyl leukotrienes (Cys-LTs) and other inflammatory markers in exhaled breath condensate (EBC). METHODS: EBC was collected, and the fraction of exhaled nitric oxide (FE NO ) was measured in a group of 19 asthmatic children, after which they performed a treadmill exercise test. Fourteen healthy children were enrolled as control subjects. RESULTS: The asthmatic children were divided into the EIB group (decrease in FEV 1 , > or =12%) and the non-EIB group. The EBC was analyzed for the presence of Cys-LTs, leukotriene B 4 , and ammonia. Asthmatic patients with EIB (mean FEV 1 decrease, 23% +/- 3%) had higher Cys-LT concentrations than either asthmatic patients without EIB or control subjects (42.2 pg/mL [median] vs 11.7 pg/mL and 5.8 pg/mL; P < 05 and P < .001, respectively). Ammonia concentrations were lower in both the EIB and non-EIB groups than in control subjects (253.2 microM and 334.6 microM vs 798.4 microM; P < .01 and P < .05, respectively). No difference in EBC leukotriene B 4 levels was found among the 3 groups. Both asthmatic groups had higher FE NO levels than control subjects ( P < .001). EBC Cys-LT ( P < .01; r = 0.7) and FE NO ( P < .05; r = 0.5) values both correlated significantly with the postexercise FEV 1 decrease. CONCLUSION: this study shows that EBC Cys-LT values are higher in asthmatic children with EIB and correlate with the decrease in FEV 1 after exercise. These findings suggest that the pathways of both Cys-LT and nitric oxide are involved in the pathogenesis of EIB.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15805996 !0091-6749 (Print) Journal Article15805996UDepartment of Pediatrics, University of Padua, Via Giustiniani 3, 35128 Padua, Italy. ~?F/Schmidt, S. M. Muller, C. E. Wiersbitzky, S. K.2005Inverse association between Chlamydia pneumoniae respiratory tract infection and initiation of asthma or allergic rhinitis in children137-44Pediatr Allergy Immunol162Adolescent Animals Asthma/*epidemiology/immunology Child Child, Preschool Chlamydophila Infections/*immunology Chlamydophila pneumoniae Cohort Studies Dermatitis, Atopic/epidemiology/immunology Dinoprost/*analogs & derivatives/analysis Female Humans Immunoenzyme Techniques Immunoglobulin A/analysis Leukotriene B4/analysis Male Pneumonia, Bacterial/*immunology Polymerase Chain Reaction Research Support, Non-U.S. Gov't Respiratory Function Tests Rhinitis/*epidemiology/immunologyMarTo evaluate the role of Chlamydia pneumoniae respiratory tract infection on pediatric asthma, allergic rhinitis or atopic eczema initiation, children of three age groups (n=1211) were prospectively studied for a C. pneumoniae infection using throat swabs and polymerase chain reaction (PCR) with enzyme immunoassay (EIA) detection. Infected children (study group, SG) were examined monthly until the agent could not be detected, quantifying persistent infection. They were compared with randomly selected, non-infected children without asthma matched for age, gender and origin (control group, CG) regarding lung function and inflammatory parameters as well as initiation of allergic diseases judged by family doctor diagnosis after, in median, 22 months. At the first follow-up examination, SG children revealed a higher leukotriene B4 (median 36 pg/ml vs. 19, p=0.04) and 8-isoprostane (median 15 pg/ml vs. 12, p=0.04) in breath condensate characterizing neutrophil, agent-related inflammation and oxidative stress in the lower airways. Cysteinyl leukotrienes, important in acute allergic inflammation, were without difference. Local, anti C. pneumoniae secretory immunoglobulin A antibodies were higher in children after C. pneumoniae infection (optical density median 0.7 vs. 0.4, p=0.001) confirming PCR-EIA results. At the final examination, there was no difference in pathological lung function tests, parameters of exhaled breath condensate or eosinophilia of the nasal mucosa. Incidence of asthma (0/55 vs. 5/54, p=0.03) and allergic rhinitis [3/53 vs. 10/52, p=0.04, odds ratio and 95% confidence interval-OR 0.25 (0.06;0.98)] as well as prevalence of asthma [1/56 vs. 9/58, p=0.02, OR 0.1 (0.01;0.81)] and allergic rhinitis [6/56 vs. 16/58, p=0.03, OR 0.32 (0.11;0.88)] were lower in the SG children. There was no association in atopic eczema. Three children with persistent infection revealed a slightly higher incidence in allergic rhinitis without significance than those with single C. pneumoniae detection (1/3 vs. 2/50), however, not to the CG. In conclusion a C. pneumoniae upper respiratory tract infection may be regarded as a protective factor for childhood asthma or allergic rhinitis in a population of kindergarten and school-age children.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15787871 !0905-6157 (Print) Journal Article15787871Department of Infectious, Bronchopulmonary and Allergic Diseases, Children's and Youth Hospital, Ernst-Moritz-Arndt University, Greifswald, Germany. schmidt3@uni-greifswald.de~?G7Morice, A. Everett, C. F. Mulrennan, S. A. Chung, K. F.2005*Exhaled breath condensate in chronic cough259; author reply 259Thorax603iBreath Tests Bronchitis/physiopathology Chronic Disease Cough/*etiology Humans Hydrogen-Ion ConcentrationMarfhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15741453 60040-6376 (Print) Comment Duplicate Publication Letter15741453 ~?H@Marteus, H. Tornberg, D. C. Weitzberg, E. Schedin, U. Alving, K.2005kOrigin of nitrite and nitrate in nasal and exhaled breath condensate and relation to nitric oxide formation219-25Thorax603OAdult Aged Anti-Infective Agents, Local/pharmacology Breath Tests Chlorhexidine/pharmacology Female Humans Male Middle Aged Mouthwashes/pharmacology Nitrates/administration & dosage/*analysis/metabolism Nitric Oxide/*metabolism Nitrites/*analysis/metabolism Nose/chemistry Research Support, Non-U.S. Gov't Saliva/chemistry TracheostomyMarBACKGROUND: Raised concentrations of nitrate and nitrite have been found in exhaled breath condensate (EBC) in airway disease, and it has been postulated that this reflects increased nitric oxide (NO) metabolism. However, the chemical and anatomical origin of nitrate and nitrite in the airways has not yet been sufficiently studied. METHODS: The fraction of exhaled NO at an exhalation flow rate of 50 ml/s (FE(NO)) and nitrite and nitrate in EBC, nasal condensate, and saliva were measured in 17 tracheostomised and 15 non-tracheostomised subjects, all of whom were non-smokers without respiratory disease. Tracheal and oral samples were taken from the tracheostomised subjects and nasal (during velum closure) and oral samples from the non-tracheostomised subjects. Measurements were performed before and after sodium nitrate ingestion (10 mg/kg) and use of antibacterial mouthwash (chlorhexidine 0.2%). RESULTS: In tracheostomised subjects oral FE(NO) increased by 90% (p<0.01) while tracheal FE(NO) was not affected 60 minutes after nitrate ingestion. Oral EBC nitrite levels were increased 23-fold at 60 minutes (p<0.001) whereas the nitrite levels in tracheal EBC showed only a minor increase (fourfold, p<0.05). Nitrate was increased the same amount in oral and tracheal EBC at 60 minutes (2.5-fold, p<0.05). In non-tracheostomised subjects oral FE(NO) and EBC nitrite increased after nitrate ingestion and after chlorhexidine mouthwash they approached baseline levels again (p<0.001). Nasal NO, nitrate, and nitrite were not affected by nitrate intake or mouthwash. At baseline, mouthwash with deionised water did not affect nitrite in oral EBC or saliva, whereas significant reductions were seen after antibacterial mouthwash (p<0.05 and p<0.001, respectively). CONCLUSIONS: Besides the salivary glands, plasma nitrate is taken up by the lower airways but not the nasal airways. Nitrate levels in EBC are thus influenced by dietary intake. Nitrate is reduced to nitrite by bacterial activity which takes place primarily in the oropharyngeal tract of healthy subjects. Only oropharyngeal nitrite seems to contribute to exhaled NO in non-inflamed airways, and there is also a substantial contribution of nitrite from the oropharyngeal tract during standard collection of EBC.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15741439 !0040-6376 (Print) Journal Article15741439^Department of Physiology and Pharmacology, Karolinska Institutet, SE-171 77 Stockholm, Sweden.~?I Horvath, I.2005yThe exhaled biomarker puzzle: bacteria play their card in the exhaled nitric oxide-exhaled breath condensate nitrite game179-80Thorax603iAsthma/*diagnosis Biological Markers/analysis Humans Nitric Oxide/*analysis Nitrites/*analysis RecurrenceMarfhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15741429 #0040-6376 (Print) Comment Editorial15741429~?JICarraro, S. Folesani, G. Corradi, M. Zanconato, S. Gaston, B. Baraldi, E.2005QAcid-base equilibrium in exhaled breath condensate of allergic asthmatic children476-81Allergy6043*Acid-Base Equilibrium Adolescent Ammonia Asthma/*etiology/*physiopathology Case-Control Studies Child *Exhalation Female Humans Hydrogen-Ion Concentration Hypersensitivity/*complications/*physiopathology Male Nitric Oxide Research Support, N.I.H., Extramural Research Support, U.S. Gov't, P.H.S. SpirometryApr BACKGROUND: The dysregulation of airway pH control may have a role in asthma pathophysiology. The measurement of exhaled breath condensate (EBC) pH and ammonia levels may be used as a noninvasive method to study acid-base status in the airway of asthmatics. METHODS: Exhaled breath condensate from 29 allergic stable asthmatic children and 13 healthy controls was collected by cooling exhaled air during tidal breathing. Ammonia was measured by high-performance liquid chromatography with fluorescence detection. pH was measured after deaeration of EBC samples by bubbling with argon. The children also underwent FENO measurement. RESULTS: Both pH and ammonia values in EBC were significantly lower in the asthmatics than in the control group [pH: ICS-treated (median and interquartile range) 7.70 (7.62-7.74), steroid-naive 7.53 (7.41-7.68), controls 7.85 (7.80-7.90), P <0.01 and P <0.001, respectively; ammonia: ICS-treated 476.17 microM (282.50-594.80), steroid-naive 253.24 microM (173.43-416.08), controls 788.30 microM (587.29-1310.39), P < 0.05 and P <0.001, respectively]. Both pH and ammonia values were higher in ICS-treated than in steroid-naive asthmatic children. There was a significant correlation between EBC pH and ammonia concentrations. CONCLUSIONS: These data show that EBC pH values of stable asthmatic children are lower compared with those of healthy controls and positively correlated with ammonia concentrations, supporting the hypothesis that airway acidification may have a role in the pathobiology of allergic asthma.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15727579 !0105-4538 (Print) Journal Article15727579>Department of Pediatrics, University of Padova, Padova, Italy. ~?KXZacharasiewicz, A. Wilson, N. Lex, C. Erin, E. M. Li, A. M. Hansel, T. Khan, M. Bush, A.2005`Clinical use of noninvasive measurements of airway inflammation in steroid reduction in children1077-82Am J Respir Crit Care Med17110JAdministration, Inhalation Adolescent Adrenal Cortex Hormones/*administration & dosage Anti-Inflammatory Agents/*administration & dosage Asthma/*complications/*drug therapy/metabolism/physiopathology Biological Markers/analysis Breath Tests Bronchial Hyperreactivity/metabolism Child Dose-Response Relationship, Drug Drug Monitoring/*methods Eosinophils/metabolism Female Follow-Up Studies Humans Inflammation/*drug therapy/*etiology/metabolism/physiopathology Male Nitric Oxide/analysis Prospective Studies Research Support, Non-U.S. Gov't Sensitivity and Specificity Sputum/metabolismMay 15The use of noninvasive methods of monitoring airway inflammation, such as exhaled nitric oxide (eNO) and induced sputum, has been shown to improve asthma monitoring and optimize treatment in adult patients with asthma. There is a lack of comparable data in children. Forty children with stable asthma eligible for inhaled steroid reduction were reviewed every 8 weeks, and their inhaled steroid dose halved if clinically indicated. eNO, sputum induction combined with bronchial hyperreactivity testing, and exhaled breath condensate collection were performed at each visit to predict success or failure of reduction of inhaled steroids. Thirty of 40 (75%) children tolerated at least one dose reduction, 12 of 40 (30%) were successfully weaned off, and in total, 15 of 40 (38%) children experienced loss of asthma control. Treatment reduction was successful in all children who had no eosinophils in induced sputum before the attempted reduction. Using multiple logistic regression, increased eNO (odds ratio, 6.3; confidence interval, 3.75-10.58) and percentage of sputum eosinophils (odds ratio, 1.38; confidence interval, 1.06-1.81) were significant predictors of failed reduction. These findings suggest that monitoring airway inflammation may be useful in optimizing treatment in children with asthma.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15709050 01073-449X (Print) Clinical Trial Journal Article15709050Department of Respiratory Pediatrics, Clinical Trials and Evaluation Unit, Royal Brompton Hospital, Sydney Street, London SW3 6NP, UK. angela.zacharasiewicz@wienkav.at ~?LCEffros, R. M. Su, J. Casaburi, R. Shaker, R. Biller, J. Dunning, M.2005aUtility of exhaled breath condensates in chronic obstructive pulmonary disease: a critical review135-9Curr Opin Pulm Med112Acidosis, Respiratory/diagnosis/etiology Breath Tests/*methods Humans Pulmonary Disease, Chronic Obstructive/complications/*diagnosis Research Support, N.I.H., Extramural Research Support, U.S. Gov't, P.H.S. Sensitivity and SpecificityMarPURPOSE OF REVIEW: Evaluation of the utility of exhaled breath condensates in chronic obstructive pulmonary disease. RECENT FINDINGS: Exhaled breath condensates have recently been introduced as a simple, noninvasive method of sampling respiratory fluid in inflammatory lung disorders, including chronic obstructive pulmonary disease. Increases in condensate concentrations of at least 12 markers of inflammation have been reported in these disorders. Furthermore, condensate pH appears to be decreased in both chronic obstructive lung disease and bronchial asthma. This has been referred to as acidopnea and could reflect airway acidification by inflammatory cells. Although safer and more convenient than bronchoalveolar lavage, interpretation of condensate data is complicated by uncertainty regarding the source of condensate solutes and by variable dilution of respiratory droplets from condensed water vapor, which represents more than 99.9% of condensate volumes. This dilution can be estimated from the dilution of plasma constituents such as urea or electrolytes. Because the principal buffer in condensate is NH4, much of which is derived from bacterial degradation of urea in the mouth, condensate pH measurements may not provide accurate estimates of airway pH. Nevertheless, acidification of condensate may be indicative of gastroesophageal reflux, which frequently occurs in obstructive lung diseases and may contribute to cough and bronchospasm. SUMMARY: It is too early to tell how useful condensate studies will be to pulmonary investigators and clinicians. Realization of the enormous potential of this approach will require a thorough understanding of the manner in which these solutions are generated and how they should be analyzed.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15699785 (1070-5287 (Print) Journal Article Review15699785]Department of Medicine, Harbor-UCLA Medical Center, Torrance, California, USA. effros@mcw.edu S~?M,Borrill, Z. Starkey, C. Vestbo, J. Singh, D.2005XReproducibility of exhaled breath condensate pH in chronic obstructive pulmonary disease269-74 Eur Respir J252(Adult Aged Biological Markers/analysis Breath Tests/*methods Case-Control Studies Exhalation Female Humans Hydrogen-Ion Concentration Male Middle Aged Pulmonary Disease, Chronic Obstructive/*diagnosis/physiopathology Reproducibility of Results Respiratory Function Tests Specimen Handling/methodsFebSIncreasingly, exhaled breath condensate (EBC) is being used to sample airway fluid from the lower respiratory tract. EBC pH may be a biomarker of airway inflammation in chronic obstructive pulmonary disease (COPD). In this study, the reproducibility of EBC pH in COPD was investigated. A total of 36 COPD patients and 12 healthy nonsmoking subjects participated in several investigations: duration of argon deaeration, within-sample variability, effect of freezing, leaving samples at room temperature, nose-peg use, within- (WD) and between-day (BD) variability. Analysis of repeated measurements was performed using the Bland-Altman method with limits of agreement (LOA; mean difference+/-2 SD). Wider LOA indicate greater variability. EBC pH became significantly higher with argon deaeration for < or =5 min. Variability during sample analysis was minimal; LOA of within-sample variability, freezing for 3 months and leaving at room temperature for 3 h were -0.29-0.45, -0.37-0.42 and -0.13-0.09, respectively. In contrast, variability due to nose-peg use (LOA -1.46-1.99), WD (LOA -1.50-2.48) and BD variability (LOA -2.52-3.02) were higher in COPD. In healthy nonsmoking subjects, nose-peg use (LOA -0.27-0.23), WD (LOA -0.33-0.40) and BD variability (LOA -0.46-0.44) were more reproducible. In conclusion, the variability of exhaled breath condensate pH in chronic obstructive pulmonary disease patients is mainly due to changes in airway pH over time, which are not seen in healthy nonsmoking subjects. Reasons for these fluctuations in exhaled breath condensate pH are unclear and require further investigation.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15684290 !0903-1936 (Print) Journal Article15684290tMedicines Evaluation Unit, Wythenshawe Hospital, North West Lung Centre, Manchester M23 9LT, UK. zborrill@meu.org.uk ~?NDGerritsen, W. B. Asin, J. Zanen, P. van den Bosch, J. M. Haas, F. J.2005jMarkers of inflammation and oxidative stress in exacerbated chronic obstructive pulmonary disease patients84-90 Respir Med991Aged Anti-Inflammatory Agents/therapeutic use Breath Tests/methods Cell Adhesion Molecules/blood Drug Monitoring/methods Female Humans Hydrogen Peroxide/metabolism Inflammation Mediators/*blood Intercellular Adhesion Molecule-1/blood Interleukin-8/blood Male Middle Aged *Oxidative Stress Prednisolone/therapeutic use Pulmonary Disease, Chronic Obstructive/drug therapy/*metabolism/physiopathologyJanCOPD is characterised by damage to small airways due to an inflammatory process as well as an imbalance between oxidants and antioxidants. Several cytokines and cell adhesion molecules enhancing a mainly neutrophilic inflammation have been associated with COPD. The aim of the study was to investigate whether inflammation or oxidative markers gave an indication of the course of COPD during an exacerbation. Fourteen patients with moderate to severe COPD admitted to the St. Antonius Hospital because of an exacerbation have been monitored during treatment with prednisolone 50 mg intravenously during 24 h at admission, reduced to 25 mg at day 3 and tapered off with oral prednisolone at day 7. On three separate occasions, day 1, 3 and 7, H2O2 in exhaled air, IL-8 and the soluble cell adhesion molecule sICAM and sE-selectin in serum were measured. We compared the patients at day 1 with healthy controls (in both non-smokers and smokers). Furthermore, we examined the changes from the study group in time during therapy. At admission all the markers were raised in comparison with the control groups. During treatment H2O2 concentrations in breath condensate declined significantly (P<0.001) as well as IL-8 and sICAM in serum (P=0.002, respectively, P<0.001). There was no significant change in sE-selectin (P=0.132). No significant improvement has been found in spirometry. These data suggest that the markers H2O2 in exhaled air, IL-8 and sICAM in serum are suitable markers in monitoring exacerbated COPD.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15672854 !0954-6111 (Print) Journal Article15672854Department of Clinical Chemistry, St. Antonius Hospital Nieuwegein, P.O. Box 2500, 3430 EM Nieuwegein, The Netherlands. w.gerritsen@kcl-azn.demon.nl |~?O1Sanak, M. Kielbasa, B. Bochenek, G. Szczeklik, A.2004JExhaled eicosanoids following oral aspirin challenge in asthmatic patients1899-904Clin Exp Allergy3412Adult Anti-Inflammatory Agents, Non-Steroidal/*adverse effects/diagnostic use Area Under Curve Aspirin/*adverse effects/diagnostic use Asthma/*chemically induced/diagnosis Biological Markers/analysis/blood/urine Breath Tests Case-Control Studies Comparative Study Dinoprost/analysis/blood Dinoprostone/analysis Eicosanoids/*analysis Female Humans Leukotriene E4/urine Lung/drug effects/*metabolism Male Mass Fragmentography Middle Aged ROC CurveDecBACKGROUND: Biochemical analysis of expiratory breath condensate is an emerging non-invasive technique for assessment of airway inflammation. OBJECTIVE: We wondered whether application of expiratory breath condensate could facilitate diagnosis of aspirin-intolerant asthma and reproduce eicosanoids mediators' abnormalities described in this disease. METHODS: We measured prostaglandins (PGs) E(2), F(2 alpha), 9 alpha 11 beta F(2) and iso-F(2) by gas-chromatography/mass-spectrometry and cysteinyl leukotrienes (cys-LTs) by radioimmunoassay in breath condensates of asthmatic patients undergoing oral aspirin challenge. Fourteen patients with aspirin-induced asthma and 20 aspirin-tolerating asthmatics, most of them on chronic inhaled corticotherapy, were studied and compared with 10 healthy subjects. Additionally, plasma 9 alpha 11 beta PGF(2), the metabolite of PGD(2) and urinary leukotriene (LT) E(4) were measured before and following the challenge. RESULTS: At baseline, PG did not differ between the groups, except for lower 9 alpha 11 beta PGF(2) in aspirin-intolerant asthma. Their concentrations were not changed by the challenge. Breath condensate cys-LTs were similar in the groups studied at base, and after aspirin challenge increased only in aspirin-intolerant patients. Elevated baseline urinary LTE(4) and its further increase following aspirin challenge was highly diagnostic for aspirin-intolerant asthma. The discriminatory value of cys-LTs increase in breath condensates was lower (72.8%) than either basal (99%) or post-challenge increase (94%) of urinary LTE(4). CONCLUSIONS: In asthmatic patients on chronic corticotherapy measurement of urinary LTE(4) excretion rather than cys-LTs in breath condensate is of greater value for diagnosis of aspirin hypersensitivity.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15663565 !0954-7894 (Print) Journal Article15663565[Department of Medicine, School of Medicine, Jagiellonian University, 31-066 Cracow, Poland. h~?P&Cap, P. Pehal, F. Chladek, J. Maly, M.2005_Analysis of exhaled leukotrienes in nonasthmatic adult patients with seasonal allergic rhinitis171-6Allergy602Adolescent Adult Case-Control Studies *Exhalation Female Humans *Leukotriene B4 *Leukotriene E4 Male Middle Aged Osmolar Concentration *Pollen Research Support, Non-U.S. Gov't Rhinitis, Allergic, Seasonal/*physiopathology *SeasonsFebBACKGROUND: Leukotrienes (LTs) are increased in exhaled breath condensate (EBC) in patients with asthma. So far no data have been reported about LT levels in nonasthmatic patients with seasonal allergic rhinitis (SAR). The aim of the study was to find out whether the LT levels in EBC were increased in the nonasthmatic adult patients with SAR both during and after the pollen season in comparison with healthy controls and to assess the changes of the LT levels after the pollen season. METHODS: Twenty-nine nonasthmatic adult patients with SAR underwent measurement of exhaled LTs in the EBC during and after the pollen season. Leukotrienes B(4), C(4), D(4) and E(4) were analysed by a specific and sensitive gas chromatography/mass spectrometry (GC/MS) assay and compared with 50 healthy nonsmoking controls. Spirometry, skin prick tests and nonspecific IgE were evaluated. RESULTS: Leukotrienes concentrations (B(4), E(4) but not D(4)) were significantly increased in and after the pollen season in patients with SAR in comparison with healthy controls. In most of the samples, LT C(4) was undetectable. The values of all exhaled LTs were significantly decreased after the pollen season compared with the seasonal baseline: LTB(4) (P = 0.023), LTD(4) (P = 0.020), LTE(4) (P = 0.047). CONCLUSIONS: Levels of exhaled LTB(4) and LTE(4) were higher in SAR patients than in healthy controls and decreased after the pollen season as compared with levels in season. The SAR patients with the highest in season LT levels had also the post-season levels elevated and this may be an early marker of inflammatory process in the lower airways despite the absence of clinical symptoms of asthma.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15647037 !0105-4538 (Print) Journal Article15647037Department of Allergology and Clinical Immunology, Hospital Na Homolce, Institute for Postgraduate Medical Education, Prague, Czech Republic.v~?Q Dwyer, T. M.2004NSampling airway surface liquid: non-volatiles in the exhaled breath condensate241-50Lung1824Albumins/metabolism Ammonia/metabolism Breath Tests/*methods Exhalation/*physiology Female Humans Male Proteins/metabolism Reference Values Research Support, Non-U.S. Gov't Respiratory Function Tests Specimen Handling/methods Surface Properties Urea/*metabolismExhaled breath condensate (EBC) samples contain molecules that have no appreciable vapor pressure; such molecules likely derive from droplets of airway fluid. We analyzed EBC gathered from a total of 62 healthy volunteers in order to quantify the volume of airway liquid that was the source of the non-volatiles; saliva was analyzed as a reference secretion. EBC urea averaged 0.52 +/- 0.12 micromol/L (n = 18), an 8,600-fold dilution from predicted blood urea nitrogen levels. Protein averaged 2.3 +/- 0.3 microg/ml (n = 31), three orders of magnitude less than in saliva (1.4 +/- 0.1 mg/ml, n = 15). EBC ammonia was 6.6 +/- 0.6 mmol/L (1/15 that of saliva) and EBC ammonium ion was 0.90 +/- 0.19 micromol/L, concentrations that are incompatible with an 8,600-fold dilution from a biological source. Thus, urea-derived dilution factors may be used to interpret EBC non-volatile molecules, but not EBC volatiles.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15636196 00341-2040 (Print) Clinical Trial Journal Article15636196Department of Physiology and Biophysics, University of Mississippi Medical Center, 2500 North State Street, Jackson, MS 30216-4505, USA. tdwyer@physiology.umsmed.edu~?RCap, P. Pehal, F.2004m[Examination of exhaled breath condensate in patients with asthma and chronic obstructive pulmonary diseases]742-6; discussion 746-7 Cas Lek Cesk14311Asthma/*diagnosis Biological Markers/analysis *Breath Tests/methods English Abstract Humans Inflammation Mediators/analysis Pulmonary Disease, Chronic Obstructive/*diagnosis Research Support, Non-U.S. Gov'tConsiderable interest of specialists all over the world has focused on the measurement of the markers of inflammation and oxidative stress in the exhaled breath condensate in patients with asthma or chronic obstructive pulmonary diseases recently. Use of exhaled condensate is based on the hypothesis that aerosol particles exhaled in human breath reflect the composition of the bronchoalveolar extracellular lining fluid. The standard collection of the material requires condensation of exhaled air and the samples have to be kept in biologically inert containers. Measurement of the very low concentrations of selected substances requires very sensitive analytical methods. The examination of exhaled breath condensate is absolutely non-invasive method, which can be repeated as often as needed and it is extremely well tolerated both by children and seniors. Markers in the condensate enable detection and quantification of the inflammation process, the disease monitoring, and assessment of the response to the treatment. The breath condensate diagnostics is a new progressive method and in the patients with asthma and chronic obstructive pulmonary disease it can bring complementary information to the very sensitive method of determination of exhaled nitric oxide.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15628568 (0008-7335 (Print) Journal Article Review15628568hVysetreni kondenzatu vydechovaneho vzduchu u nemocnych s astmatem a chronickou obstrukcni plicni nemoci.[Oddeleni alergologie a klinicke imunologie Nemocnice Na Homolce, Praha. petr.cap@homolka.cz~?SVWyse, C. A. Hammond, J. Arteaga, A. Cumming, D. Cooper, J. M. McEwan, J. D. Yam, P. S.2004`Collection and analysis of exhaled breath condensate hydrogen peroxide in conscious healthy dogs744-6Vet Rec15523Animals Breath Tests/instrumentation/*methods Dog Diseases/diagnosis Dogs/*metabolism Exhalation Hydrogen Peroxide/*analysis Research Support, Non-U.S. Gov't Respiratory Tract Diseases/diagnosis/veterinaryDec 4fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15623088 !0042-4900 (Print) Journal Article15623088Institute of Comparative Medicine, Department of Veterinary Clinical Studies, University of Glasgow Veterinary School, Bearsden, Glasgow G61 IQH. !~?TjWells, K. Vaughan, J. Pajewski, T. N. Hom, S. Ngamtrakulpanit, L. Smith, A. Nguyen, A. Turner, R. Hunt, J.2005FExhaled breath condensate pH assays are not influenced by oral ammonia27-31Thorax601RAdult Aged Ammonia/*analysis Biological Markers Breath Tests/*methods Bronchi/*metabolism Female Freeze Drying Humans Hydrogen-Ion Concentration Male Middle Aged Mouthwashes Reproducibility of Results Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Sensitivity and Specificity Spectrophotometry/standards TemperatureJanBACKGROUND: Measurement of pH in exhaled breath condensate (EBC) is robust and simple. Acidic source fluid (airway lining fluid) traps bases while volatilising acids, leading to EBC acidification in many lung diseases. Lower airway ammonia is one determinant of airway lining fluid pH, raising the concern that addition of the base ammonia by contamination from the mouth might confound EBC pH assays. METHODS: Three discrete methods were used to limit oral ammonia contamination of EBC collections: endotracheal intubation, oral rinsing, and -40 degrees C condenser temperatures. Separately, ammonia was removed from collected EBC samples by lyophilisation and resuspension. Intraweek and intraday variability of ammonia concentration was determined in 76 subjects, and ammonia and pH from a further 235 samples were graphically compared. Ammonia was assayed spectrophotometrically and pH was assessed after deaeration. RESULTS: Data from 1091 samples are presented. Ammonia was reduced in EBC by all methods. Endotracheal intubation decreased EBC ammonia from a mean (SD) of 619 (124) microM to 80 (24) microM (p<0.001, n=32). Oral rinsing before collection also led to a decline in EBC ammonia from 573 (307) microM to 224 (80) microM (p=0.016, n=7). The colder the condensation temperature used, the less ammonia was trapped in the EBC. Lyophilisation removed 99.4 (1.9)% of ammonia. Most importantly, the pH of EBC never decreased after removal of ammonia by any of these methods. Intraweek and intraday coefficients of variation for ammonia were 64 (27)% and 60 (32)%, which is substantially more variable than EBC pH assays. CONCLUSIONS: Although ammonia and pH appear to correlate in EBC, the oral ammonia concentration is not an important determinant of EBC pH. No precautions need to be taken to exclude oral ammonia when EBC pH is of interest. The low pH and low ammonia found in EBC from patients with lung diseases appear to be independent effects of volatile compounds arising from the airway.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15618579 !0040-6376 (Print) Journal Article15618579oDivision of Pediatric Respiratory Medicine, Box 800386, University of Virginia, Charlottesville, VA 22908, USA.!~?UKOjoo, J. C. Mulrennan, S. A. Kastelik, J. A. Morice, A. H. Redington, A. E.2005_Exhaled breath condensate pH and exhaled nitric oxide in allergic asthma and in cystic fibrosis22-6Thorax601Adult Asthma/*metabolism Breath Tests/methods Case-Control Studies Cystic Fibrosis/*metabolism Exhalation Female Forced Expiratory Volume/physiology Humans Hydrogen-Ion Concentration Male Nitric Oxide/analysis/*metabolism Respiratory Mucosa/metabolism Skin TestsJan=BACKGROUND: It has been proposed that the pH of airway lining fluid may regulate the fractional exhaled concentration of nitric oxide (Fe(NO)) in respiratory disease. METHODS: Fe(NO), exhaled breath condensate (EBC) pH, and EBC concentrations of nitrite plus nitrate (NO2/NO3) were compared in 12 subjects with stable asthma, 18 with stable cystic fibrosis (CF), and 15 healthy control subjects. Eight of the CF patients were studied on a separate occasion at the start of a pulmonary exacerbation. RESULTS: Fe(NO) was significantly greater in asthmatic subjects than in control subjects (mean 35 v 9 ppb, p<0.001). EBC pH, however, was similar in the asthmatic and control groups (median 5.82 v 6.08, p=0.23). Levels of NO2/NO3 were on average higher in EBC samples from asthmatic subjects, but the difference was not significant. In patients with stable CF both the Fe(NO) (mean 4 ppb, p<0.001) and EBC pH (median 5.77, p=0.003) were lower than in the control group. Levels of EBC NO2/NO3 (median 29.9 microM; p=0.002) in patients with stable CF, in contrast, were significantly higher than in control subjects. During CF exacerbations, EBC pH was further reduced (median 5.30, p=0.017) but Fe(NO) and NO2/NO3 were unchanged. CONCLUSIONS: These findings demonstrate a dissociation between EBC pH and Fe(NO) in inflammatory airways disease.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15618578 !0040-6376 (Print) Journal Article15618578\Division of Academic Medicine, Postgraduate Medical Institute, University of Hull, Hull, UK.:~?VHarrison, C. M. Andersen, C. C.2005ZExhaled breath measures of inflammation: are they useful in neonatal chronic lung disease?F6-10 Arch Dis Child Fetal Neonatal Ed901Biological Markers/analysis Breath Tests/*methods Carbon Monoxide/analysis Chronic Disease Humans Hydrocarbons/analysis Infant, Newborn Infant, Premature Infant, Premature, Diseases/*diagnosis Lung Diseases/*diagnosis Nitric Oxide/analysis Pneumonia/diagnosisJanYNeonatal chronic lung disease is a common problem for surviving infants of extreme prematurity. Although the precise pathophysiology is still not known, it is clear that inflammation provides a common link that amplifies the injury to the premature lung. Current invasive measures of pulmonary inflammation include markers in blood and airway effluent, with the cellular composition of tracheal fluid being the "gold standard". In this article available exhaled breath measures, particularly nitric oxide, carbon monoxide, volatile hydrocarbons, and exhaled breath condensate, are reviewed with particular reference to sample collection, analysis, and common pitfalls as they apply to the ventilated premature newborn at risk of chronic lung disease. Although they have great potential, all measures require thorough validation before being used clinically.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15613578 (1359-2998 (Print) Journal Article Review15613578kDepartment of Pediatrics, Mercy Hospital for Women, Clarendon St, East Melbourne, Victoria 3002, Australia.X~?WASvensson, S. Isacsson, A. C. Ljungkvist, G. Toren, K. Olin, A. C.2005Optimization and validation of an ion chromatographic method for the simultaneous determination of sodium, ammonium and potassium in exhaled breath condensate173-7-J Chromatogr B Analyt Technol Biomed Life Sci8141*Breath Tests Calibration Chromatography, Liquid/*methods Humans Potassium/*analysis Quaternary Ammonium Compounds/*analysis Reproducibility of Results Sensitivity and Specificity Sodium/*analysisJan 5An ion chromatographic method with conductivity detection for the simultaneous quantification of sodium, ammonium and potassium in exhaled breath condensate (EBC) was developed and validated. A factorial design was used to optimize the chromatographic conditions, which resulted in baseline separations of the cations within 6 min. The method requires no pre-treatment of EBC samples. The optimized method was used for the intra-individual screening of cations in EBC of 10 healthy volunteers. The LOQs were low (0.3, 0.1 and 0.2 microM for sodium, ammonium and potassium, respectively), compared with levels detected in healthy volunteers. The responses were linear with good precision, and samples could be stored for at least 10 weeks at refrigerating conditions.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15607722 41570-0232 (Print) Journal Article Validation Studies15607722Occupational and Environmental Medicine, Sahlgrenska University Hospital and Academy at Goteborg University, SE-41266 Goteborg, Sweden. sophie.svensson@ymk.gu.se ~?XjDe Benedetto, F. Aceto, A. Dragani, B. Spacone, A. Formisano, S. Pela, R. Donner, C. F. Sanguinetti, C. M.2005PLong-term oral n-acetylcysteine reduces exhaled hydrogen peroxide in stable COPD41-7Pulm Pharmacol Ther181Acetylcysteine/administration & dosage/*therapeutic use *Administration, Oral Aged Breath Tests/instrumentation/methods Comparative Study Drug Administration Schedule Exhalation/*drug effects/physiology Female Humans Hydrogen Peroxide/chemistry/*metabolism Male Pulmonary Disease, Chronic Obstructive/*drug therapy/etiology/physiopathology Spirometry/instrumentation/methods Time FactorsOxidative stress caused by airway inflammation is increased in chronic obstructive pulmonary disease (COPD) and may account for the progressive deterioration of structure and function of the respiratory tract observed in this disease. Antioxidant defences of the respiratory tract may be overwhelmed by the oxidant burden in COPD and possibly restored with antioxidant therapy. The level of hydrogen peroxide (H(2)O(2)) concentration in exhaled air condensate (EAC) is a valuable tool for assessing and monitoring oxidative stress. This study aimed to verify the effect of 2-month oral N-acetylcysteine (NAC) treatment compared to placebo on the H(2)O(2) content in EAC of 55 clinically stable COPD patients (48 males), mean age 65.93+/-9.3 years. After clinical examination, pulmonary function tests, and collection of EAC for the basal (T0) assay of H(2)O(2), patients were randomly allocated to group A (usual therapy plus oral NAC 600 mg b.i.d. for 2 months) or group B (usual therapy plus placebo b.i.d. for 2 months). H(2)O(2) assay in EAC was repeated at 15 (T15), 30 (T30), and 60 (T60) days after the start of therapy in each group. All patients were non-smokers or ex smokers for at least 5 years and the two groups were comparable in terms of demographic, respiratory function, and EAC data at baseline. The H(2)O(2) level in EAC of group A was significantly decreased at T15 (1.00+/-0.38 SD microM; p=0.003), T30 (0.91+/-0.44 microM; p=0.007), and T60 (0.83+/-0.41 microM; p=0.000) compared to T0 (1.28+/-0.61 microM). No significant decrease in H(2)O(2) of group B was found at any time point. We conclude that oral NAC 600 mg b.i.d. for 2 months rapidly reduces the oxidant burden in airways of stable COPD patients.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15607126 L1094-5539 (Print) Clinical Trial Journal Article Randomized Controlled Trial15607126wDepartment of Pneumology, Ospedale San Camillo De Lellis, Via Carlo Forlanini, 50 66100 Chieti, Italy. debened@unich.it V~?YfCorradi, M. Pignatti, P. Manini, P. Andreoli, R. Goldoni, M. Poppa, M. Moscato, G. Balbi, B. Mutti, A.2004`Comparison between exhaled and sputum oxidative stress biomarkers in chronic airway inflammation1011-7 Eur Respir J246Adult Aged Aged, 80 and over Aldehydes/*analysis Asthma/*metabolism/physiopathology Biological Markers/*analysis Breath Tests Case-Control Studies Chromatography, Liquid Exhalation Female Humans Lipid Peroxidation Male Middle Aged Oxidative Stress Pulmonary Disease, Chronic Obstructive/*metabolism/physiopathology Research Support, U.S. Gov't, P.H.S. Spectrum Analysis, Mass Sputum/*metabolism Statistics, NonparametricDecThe aim of the present study was to compare aldehyde levels resulting from lipid peroxidation in exhaled breath condensate (EBC) and induced sputum (IS) supernatant of subjects with asthma and chronic obstructive pulmonary disease (COPD). Aldehydes (malondialdehyde (MDA), acrolein, n-hexanal (C6), n-heptanal (C7), n-nonanal (C9), 4-hydroxynonenal (HNE) and 4-hydroxyhexenal (HHE)) in both biological fluids were measured by liquid chromatography-tandem mass spectrometry. MDA concentrations in sputum were 132.5 nM (82.5-268.8) and 23.7 nM (9-53.7) in EBC. Similarly, C6, C7 and C9 concentrations in IS were 1.5-4.7-fold higher than in EBC. Acrolein levels were 131.1 nM (55.6-264.6) in IS and 45.3 nM (14.4-127.1) in EBC. The concentrations of HNE and HHE in IS were not significantly different from the levels in EBC. Aldehyde levels in EBC did not show any correlation with aldehyde levels in IS or with differential sputum cellular count. In COPD, MDA in EBC, but not its IS counterpart, was negatively correlated with the severity of disease. In conclusion, the data presented here show that aldehydes can be detected in both exhaled breath condensate and supernatant of induced sputum, but that their relative concentrations are different and not correlated with each other. Therefore, with regard to lipid peroxidation products, exhaled breath condensate and induced sputum must be considered as independent techniques.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15572547 !0903-1936 (Print) Journal Article15572547xNational Institute of Occupational Safety and Prevention Research Center at the University of Parma, 43100 Parma, Italy. ,~?ZcCarpagnano, G. E. Kharitonov, S. A. Foschino-Barbaro, M. P. Resta, O. Gramiccioni, E. Barnes, P. J.2004oSupplementary oxygen in healthy subjects and those with COPD increases oxidative stress and airway inflammation1016-9Thorax5912;Bronchitis/*etiology Female Forced Expiratory Volume/physiology Humans Immunoassay Interleukin-6/analysis Isoprostanes/analysis Male Middle Aged Oxidative Stress/*physiology Oxygen Inhalation Therapy/*methods Pulmonary Disease, Chronic Obstructive/*therapy Research Support, Non-U.S. Gov't Vital Capacity/physiologyDecBACKGROUND: Hyperoxia increases oxidative stress through the generation of reactive oxygen species and may therefore enhance inflammation in the lungs. The aim of this study was to investigate whether short term supplementary oxygen (28%) increases oxidative stress and inflammation in the airways by measuring 8-isoprostane and interleukin 6 (IL-6) concentrations in exhaled breath condensate. METHODS: Twenty three healthy subjects (12 men, mean (SD) age 48 (7) years) and 23 patients with chronic obstructive pulmonary disease (COPD; 15 men, mean (SD) age 56 (5) years) were studied. 8-isoprostane and IL-6 concentrations were measured by immunoassay. RESULTS: Increased concentrations of 8-isoprostane and IL-6 were found in all subjects after breathing 28% oxygen for 1 hour. In healthy subjects the concentrations of 8-isoprostane and IL-6 were 10.9 (2.9) pg/ml and 4.9 (0.8) pg/ml, respectively, compared with baseline concentrations of 6.1 (1.3) pg/ml and 2.9 (0.6) pg/ml, and in patients with COPD the concentrations were 27.9 (3.1) pg/ml and 8.3 (1.2) pg/ml), respectively, compared with baseline concentrations of 18.9 (3.6) pg/ml and 6.3 (0.6) pg/ml. By contrast, breathing air through the same face mask for 1 hour had no significant effects on 8-isoprostane or IL-6 concentrations in normal subjects or those with COPD. CONCLUSIONS: These findings suggest that short term supplementary oxygen may enhance oxidative stress and inflammation in the airways. Whether this happens with long term oxygen therapy needs to be determined.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15563698 !0040-6376 (Print) Journal Article15563698=Institute of Respiratory Diseases, University of Bari, Italy. ~?[yCarpagnano, G. E. Resta, O. Foschino-Barbaro, M. P. Spanevello, A. Stefano, A. Di Gioia, G. Serviddio, G. Gramiccioni, E.2004Exhaled Interleukine-6 and 8-isoprostane in chronic obstructive pulmonary disease: effect of carbocysteine lysine salt monohydrate (SCMC-Lys)169-75Eur J Pharmacol5051-3{Adult Carbocysteine/administration & dosage/*analogs & derivatives/*therapeutic use Comparative Study Dinoprost/*analogs & derivatives/*metabolism Exhalation Female Humans Interleukin-6/*metabolism Male Middle Aged Pulmonary Disease, Chronic Obstructive/*drug therapy/metabolism/pathology Research Support, Non-U.S. Gov't Respiratory Function Tests Time Factors Treatment OutcomeNov 28Chronic obstructive pulmonary disease (COPD) is characterized by an airways inflammation and by an enhanced generation of reactive oxygen species. The aim of our study was to assess the inflammation and the oxidative stress in airways of COPD patients with acute exacerbation of disease and in stability. Furthermore, we investigated the anti-inflammatory and antioxidant effects of 6 months treatment with carbocysteine lysine salt monohydrate (SCMC-Lys) in COPD. We studied 30 mild acute COPD, 10 mild stable COPD and 15 healthy subjects. 8-isoprostane and Interleukine-6 were measured in their breath condensate through immunoassay. Significantly higher concentrations of exhaled 8-isoprostane and Interleukine-6 were found in acute COPD patients compared to stable COPD and healthy controls (21.8+/-5.1 vs. 13.2+/-2.0 vs. 4.7+/-1.8 pg/ml and 7.4+/-0.9 vs. 5.8+/-0.2 vs. 2.7+/-0.6 pg/ml, p<0.0001). COPD patients treated with SCMC-Lys showed a marked reduction of exhaled 8-isoprostane and Interleukine-6 (8.9+/-1.5 and 4.6+/-0.8 pg/ml, p<0.0001). These findings suggest that there is an increase of 8-isoprostane and Interleukine-6 concentrations in the breath condensate of COPD patients compared to healthy controls especially during acute exacerbations of the disease. Moreover, we showed an anti-inflammatory and antioxidant effect of short-term administration of SCMC-Lys in COPD, suggesting the importance of a further placebo-controlled study that should evaluate the effects of this drug.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15556150 J0014-2999 (Print) Clinical Trial Controlled Clinical Trial Journal Article15556150`Institute of Respiratory Diseases, University of Medicine, Foggia, Italy. ge.carpagnano@unifg.it~?\:Doniec, Z. Nowak, D. Tomalak, W. Pisiewicz, K. Kurzawa, R.2005}Passive smoking does not increase hydrogen peroxide (H2O2) levels in exhaled breath condensate in 9-year-old healthy children41-5Pediatr Pulmonol391Biological Markers/analysis Breath Tests Child *Environmental Exposure Female Humans Hydrogen Peroxide/*analysis Male Oxidative Stress Research Support, Non-U.S. Gov't Respiratory Function Tests *Tobacco Smoke PollutionJanEnvironmental tobacco smoke, also called passive smoking, was shown to have adverse effects on the health of children. Hydrogen peroxide (H2O2) is proposed as a sensitive marker of oxidative injury and inflammatory processes in the airways, being increased in adult active cigarette smokers. We tested whether passive smoking had an influence on H2O2 exhalation in healthy children. Thirty healthy passive smoking and 24 nonexposed healthy children aged 9 years were included in the study. Exhaled breath condensate (EBC) was obtained by spontaneous tidal volume breathing with EcoScreen (Jaeger, Germany). All subjects underwent flow-volume measurements immediately after EBC collection. Levels of H2O2 were measured fluorimetrically with the homovanillic acid method. Lung function did not differ between the passive smoking and nonexposed children groups. In the passive smoking group, EBC H2O2 concentration (median and range) was 0.32 (0.00-1.20) microM, and did not differ significantly (P >0.05) from that found in the nonexposed group, i.e., 0,22 (0.00-0.68) microM. Exhaled H2O2 did not correlate with spirometric parameters (FEV1, FEV1%FVC, and MEF50%FVC) in either group. We conclude that passive smoking does not increase H2O2 exhalation in healthy children.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15532100 08755-6863 (Print) Clinical Trial Journal Article15532100YNational Institute for Tuberculosis and Lung Diseases, Branch Rabka-Zdroj, Rabka, Poland. ~?]7Effros, R. M. Dunning, M. B., 3rd Biller, J. Shaker, R.20044The promise and perils of exhaled breath condensatesL1073-80"Am J Physiol Lung Cell Mol Physiol2876Breath Tests/*methods Exhalation/*physiology Humans Lung/cytology/physiology Models, Biological Research Support, U.S. Gov't, P.H.S. Respiratory Mucosa/physiologyDecThe exhaled breath condensate (EBC) approach provides a convenient and noninvasive approach for sampling the pulmonary epithelial lining fluid (ELF). Increased EBC concentrations of more than a dozen inflammatory markers and hydrogen ions have been reported in lung diseases associated with inflammation. However, the usefulness of EBC is compromised by uncertainties concerning the sources of the EBC droplets and by the extreme and variable dilution of ELF droplets with condensed water vapor ( approximately 20,000-fold). Reported increases in EBC concentrations may reflect proportionate increases in the total volume rather than the concentration of ELF droplets in the collected samples. Conclusions regarding ELF concentrations can only be made if this dilution is estimated with a dilutional indicator (e.g., conductivity of lyophilized EBC). In normal EBC samples, pH is effectively set by oral contamination with NH(3), and EBC pH cannot provide reliable information regarding ELF pH in normal subjects. Acidification of EBC observed in asthma and other conditions may reflect acidification of ELF, decreases in NH(3) added to the EBC, and/or the presence of gastric droplets in the EBC.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15531756 (1040-0605 (Print) Journal Article Review15531756fHarbor-University of California at Los Angeles Medical Center, Torrance, CA 90502, USA. effros@mcw.edun~?^>Montuschi, P. Martello, S. Felli, M. Mondino, C. Chiarotti, M.2004oIon trap liquid chromatography/tandem mass spectrometry analysis of leukotriene B4 in exhaled breath condensate2723-9Rapid Commun Mass Spectrom1822Adolescent Adult Asthma/metabolism Atmospheric Pressure *Breath Tests Child Chromatography, High Pressure Liquid/*methods Exhalation Female Humans Leukotriene B4/*analysis/metabolism Male Research Support, Non-U.S. Gov't Spectrometry, Mass, Electrospray Ionization/*methodsHThe objective of this study is the measurement of leukotriene B7 (LTB4), a potent inflammatory mediator, in exhaled breath condensate by using liquid chromatography/mass spectrometry (LC/MS and LC/MS/MS). Condensation of exhaled breath is a non-invasive method to collect airway secretions. Deuterated (d4)-LTB4 was used as internal standard. The MS and MS/MS behavior of LTB4 and LTB4-d4 was studied by electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI) in both positive and negative ion polarity mode. Preliminary results show that monitoring negative ions in ESI mode has the best sensitivity for both LTB4 and LTB4-d4. Therefore, negative ESI was chosen, and the [M-H]- ions at m/z 335 and 339 were selected for quantification. The lower limit of quantification for LTB4, expressed as the lowest point of the calibration curve, was 100 pg/mL. Using this technique, we measured LTB4 in exhaled breath condensate in two healthy subjects, four asthmatic patients on anti-inflammatory treatment, and four asthmatic patients who were not on anti-inflammatory drugs. Exhaled LTB4 concentrations were detected only in asthmatic patients who were not on anti-inflammatory therapy. This method is potentially useful for non-invasive assessment of airway inflammation, but the sensitivity of the technique needs to be improved.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15499663 !0951-4198 (Print) Journal Article15499663Department of Pharmacology, Faculty of Medicine, Catholic University of the Sacred Heart, Rome, Italy. pmontuschi@rm.unicatt.it3~?_BNeubauer, B. Schotte, U. Struck, N. Langfeldt, N. Mutzbauer, T. S.2004[Leukotriene-B4 concentrations in breathing condensate before and after simulated deep dives217-24Undersea Hyperb Med312Adult Biological Markers/analysis Breath Tests Chi-Square Distribution Diving/*physiology Humans Leukotriene B4/*analysis Male Middle Aged Research Support, Non-U.S. Gov't Respiratory Function Tests SpirometrySummerDuring diving the respiratory tract is exposed to occupational hazards (increased oxygen partial pressure, pulmonary vessel engorgement during submersion, inert gas micro embolism during decompression). Leukotriene-B4 [LTB4] concentrations in the exhaled breath mirrors the inflammatory activity of the airways if the respiratory tract has been exposed to occupational hazards. In this study LTB4-concentrations in the exhaled breath and spirometry data obtained before and after simulated dives helped to elucidate any contributions by hyperbaric exposure to impaired lung function and to separate effects of ambient pressure from those of submersion and increased oxygen partial pressure. Thirty two healthy subjects carried out dives in a hyperbaric chamber using a cross over design to 600 kPa ambient pressure with and without submersion and a dry exposure to pure oxygen at 120 kPa ambient pressure (durations: 43 min). Pre-dive and four hours after surfacing the exhaled breath was collected non-invasively. Condensate was measured by a standard enzyme immuno-assay for LTB4 in parallel with lung function values (FVC, FEV1, MEF 25-75). Pre-exposure baseline values of LTB4-concentrations and lung function values were in the normal range. Post-exposure values did not differ significantly from the baseline values. The data gave no evidence of any inflammatory activity in the subjects' airways after hyperbaric exposure.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15485084 !1066-2936 (Print) Journal Article15485084FBehorde fur Umwelt und Gesundheit, Tesdorpfstrasse 8, D-20148 Hamburg. x~?`VMondino, C. Ciabattoni, G. Koch, P. Pistelli, R. Trove, A. Barnes, P. J. Montuschi, P.2004`Effects of inhaled corticosteroids on exhaled leukotrienes and prostanoids in asthmatic children761-7J Allergy Clin Immunol1144YAdministration, Inhalation Adrenal Cortex Hormones/*administration & dosage Asthma/diagnosis/*immunology Breath Tests/methods Child Cross-Sectional Studies Humans Leukotrienes/analysis/*immunology Nitric Oxide/analysis/*immunology Pilot Projects Pneumonia/diagnosis/immunology Prostaglandins/analysis/*immunology Research Support, Non-U.S. Gov'tOct'BACKGROUND: Lipid mediators play an important pathophysiologic role in atopic asthmatic children, but their role in the airways of atopic nonasthmatic children is unknown. OBJECTIVE: We sought (1) to measure leukotriene (LT) E 4 , LTB 4 , 8-isoprostane, prostaglandin E 2 , and thromboxane B 2 concentrations in exhaled breath condensate in atopic asthmatic and atopic nonasthmatic children; (2) to measure exhaled nitric oxide (NO) as an independent marker of airway inflammation; and (3) to study the effect of inhaled corticosteroids on exhaled eicosanoids. METHODS: Twenty healthy children, 20 atopic nonasthmatic children, 30 steroid-naive atopic asthmatic children, and 25 atopic asthmatic children receiving inhaled corticosteroids were included in a cross-sectional study. An open-label study with inhaled fluticasone (100 microg twice a day for 4 weeks) was undertaken in 14 steroid-naive atopic asthmatic children. RESULTS: Compared with control subjects, exhaled LTE 4 ( P <.001), LTB 4 ( P <.001), and 8-isoprostane ( P <.001) levels were increased in both steroid-naive and steroid-treated atopic asthmatic children but not in atopic nonasthmatic children (LTE 4 , P=.14; LTB 4 , P=.23; and 8-isoprostane, P=.52). Exhaled NO levels were increased in steroid-naive atopic asthmatic children ( P <.001) and, to a lesser extent, in atopic nonasthmatic children ( P <.01). Inhaled fluticasone reduced exhaled NO (53%, P <.0001) and, to a lesser extent, LTE 4 (18%, P <.01) levels but not LTB 4 , prostaglandin E 2 , or 8-isoprostane levels in steroid-naive asthmatic children. Conclusions Exhaled LTE 4 , LTB 4 , and 8-isoprostane levels are increased in atopic asthmatic children but not in atopic nonasthmatic children. In contrast to exhaled NO, these markers seem to be relatively resistant to inhaled corticosteroids.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15480313 !0091-6749 (Print) Journal Article15480313XDepartment of Immunodermatology, Istituto Dermopatico dell'Immacolata, IDI, IRCCS, Rome.f~?a3Hitka, P. Cerny, M. Vizek, M. Wilhelm, J. Zoban, P.20049Assessment of exhaled gases in ventilated preterm infants561-4 Physiol Res535 Breath Tests/*instrumentation/*methods Exhalation/physiology Humans Hydrogen Peroxide/*analysis/*metabolism Infant, Newborn Infant, Premature/*physiology Pilot Projects Pulmonary Gas Exchange/*physiology Research Support, Non-U.S. Gov't Respiration, Artificial/*methodsyHydrogen peroxide (H2O2) production in exhaled air was measured in ventilated preterm newborns at 5, 24 and 48 hours after delivery, using originally designed method of exhaled breath condensate (EBC) collection. H2O2 production in expired gas was 812+/-34 pmol/20 min during the first measurement and then declined to 389+/-21 at 24 hours and 259+/-26 pmol/20 min at 48 hours.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15479136 00862-8408 (Print) Clinical Trial Journal Article15479136Department of Pathophysiology, Charles University, Second Faculty of Medicine, Praha 5, Plzenska 221, Czech Republic. patrikhitka@hotmail.com~?b Griese, M. Noss, J. Schramel, P.2003JElemental and ion composition of exhaled air condensate in cystic fibrosis136-42 J Cyst Fibros23 Adolescent Adult Analysis of Variance Anions/*metabolism Breath Tests Child Chromatography, Ion Exchange Cystic Fibrosis/*metabolism *Elements Exhalation Female Humans Lung/*metabolism Male Respiratory Function Tests Spectrum Analysis/methods Statistics, NonparametricSepBACKGROUND: In cystic fibrosis (CF) the exact ion composition of the airway surface fluid is still debated and it is not clear if it differs from healthy subjects. The air that we exhale contains small droplets, which are generated by shear forces from the airway surface fluid and very likely mirror its ion composition. We hypothesized that differences between CF-patients and healthy controls would be reflected by differences in their exhaled air. METHODS: In nasally collected exhaled breath condensate from 20 children and young adults with cystic fibrosis and 20 healthy subjects, the elements and anions were determined by optical emission spectroscopy and ion-exchange chromatography. RESULTS: The concentrations of the major components Na and Cl- did not differ, Zn was higher and NO3- was lower in CF-patients. During a given time period, CF-patients produced a slightly larger volume of breath condensate and they exhaled more Na, K and Zn. Fluoride was detected in half of all samples, whereas copper, iron, magnesium, phosphorus and sulfur were present only sporadically, with no differences. CONCLUSIONS: These data detail the composition of exhaled breath condensate and suggest a similar Na and Cl- concentration in CF-airway surface fluid as in healthy subjects.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15463862 !1569-1993 (Print) Journal Article15463862Children's Hospital, Ludwig Maximilians-University of Munich, Lindwurmstr. 4, Munich 80337, Germany. griese@pk-i.med.uni-muenchen.de~?cBSparkes, A. H. Mardell, E. J. Deaton, C. Kirschvink, N. Marlin, D.2004yExhaled breath condensate (EBC) collection in cats--description of a non-invasive technique to investigate airway disease335-8J Feline Med Surg65Animals Biological Markers Breath Tests/*methods Case-Control Studies Cat Diseases/*diagnosis/metabolism Cats Female Hydrogen Peroxide/metabolism Leukotrienes/metabolism Lung Diseases/diagnosis/*veterinary Male Nitric Oxide/metabolism Predictive Value of Tests Prostaglandins/metabolismOct'Exhaled breath condensate has been collected in other species and used as a non-invasive method of evaluating airway disease by measurement of various markers in the fluid, including hydrogen peroxide, nitric oxide, leukotrienes and prostaglandins. We describe a novel technique for the collection of exhaled breath condensate from cats, which enabled collection of fluid and measurement of its hydrogen peroxide concentration. Further studies will be needed to establish the value of this technique in the investigation of feline respiratory disease.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15363765 41098-612X (Print) Evaluation Studies Journal Article15363765Centre for Small Animal Studies, The Animal Health Trust, Lanwades Park, Kentford, Newmarket, Suffolk, CB8 7UU, UK. andy.sparkes@aht.org.uk ~?dkGoldoni, M. Catalani, S. De Palma, G. Manini, P. Acampa, O. Corradi, M. Bergonzi, R. Apostoli, P. Mutti, A.2004xExhaled breath condensate as a suitable matrix to assess lung dose and effects in workers exposed to cobalt and tungsten1293-8Environ Health Perspect11213Adult *Breath Tests Case-Control Studies Cobalt/*analysis/urine Environmental Monitoring Female Humans Male *Occupational Exposure Oxidative Stress Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Sensitivity and Specificity Tungsten/*analysis/urineSepThe aim of the present study was to investigate whether exhaled breath condensate (EBC), a fluid formed by cooling exhaled air, can be used as a suitable matrix to assess target tissue dose and effects of inhaled cobalt and tungsten, using EBC malondialdehyde (MDA) as a biomarker of pulmonary oxidative stress. Thirty-three workers exposed to Co and W in workshops producing either diamond tools or hard-metal mechanical parts participated in this study. Two EBC and urinary samples were collected: one before and one at the end of the work shift. Controls were selected among nonexposed workers. Co, W, and MDA in EBC were analyzed with analytical methods based on mass spectrometric reference techniques. In the EBC from controls, Co was detectable at ultratrace levels, whereas W was undetectable. In exposed workers, EBC Co ranged from a few to several hundred nanomoles per liter. Corresponding W levels ranged from undetectable to several tens of nanomoles per liter. A parallel trend was observed for much higher urinary levels. Both Co and W in biological media were higher at the end of the work shift in comparison with preexposure values. In EBC, MDA levels were increased depending on Co concentration and were enhanced by coexposure to W. Such a correlation between EBC MDA and both Co and W levels was not observed with urinary concentration of either element. These results suggest the potential usefulness of EBC to complete and integrate biomonitoring and health surveillance procedures among workers exposed to mixtures of transition elements and hard metals. Key words: cobalt, exhaled breath condensate, hard metals, lung, malondialdehyde, oxidative stress, tungsten.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15345342 !0091-6765 (Print) Journal Article15345342sNational Institute of Occupational Safety and Prevention, Research Centre at the University of Parma, Parma, Italy. I~?eRahman, I. Biswas, S. K.2004VNon-invasive biomarkers of oxidative stress: reproducibility and methodological issues125-43 Redox Rep93Antioxidants/analysis Biological Markers Breath Tests/*methods Humans Inflammation/metabolism Lipid Peroxidation *Lung Diseases/metabolism Oxidants/analysis *Oxidative Stress Reactive Nitrogen Species/metabolism Reactive Oxygen Species/metabolism *Reproducibility of ResultsAOxidative stress is the hallmark of various chronic inflammatory lung diseases. Increased concentrations of reactive oxygen species (ROS) in the lungs of such patients are reflected by elevated concentrations of oxidative stress markers in the breath, airways, lung tissue and blood. Traditionally, the measurement of these biomarkers has involved invasive procedures to procure the samples or to examine the affected compartments, to the patient's discomfort. As a consequence, there is a need for less or non-invasive approaches to measure oxidative stress. The collection of exhaled breath condensate (EBC) has recently emerged as a non-invasive sampling method for real-time analysis and evaluation of oxidative stress biomarkers in the lower respiratory tract airways. The biomarkers of oxidative stress such as H2O2, F2-isoprostanes, malondialdehyde, 4-hydroxy-2-nonenal, antioxidants, glutathione and nitrosative stress such as nitrate/nitrite and nitrosated species have been successfully measured in EBC. The reproducibility, sensitivity and specificity of the methodologies used in the measurements of EBC oxidative stress biomarkers are discussed. Oxidative stress biomarkers also have been measured for various antioxidants in disease prognosis. EBC is currently used as a research and diagnostic tool in free radical research, yielding information on redox disturbance and the degree and type of inflammation in the lung. It is expected that EBC can be exploited to detect specific levels of biomarkers and monitor disease severity in response to appropriate prescribed therapy/treatment.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15327743 (1351-0002 (Print) Journal Article Review15327743Department of Environmental Medicine, Division of Lung Biology and Disease Program, University of Rochester Medical Center, Rochester, USA. irfan_rahman@urmc.rochester.eduT~?f=Svensson, S. Olin, A. C. Larstad, M. Ljungkvist, G. Toren, K.2004vDetermination of hydrogen peroxide in exhaled breath condensate by flow injection analysis with fluorescence detection199-203-J Chromatogr B Analyt Technol Biomed Life Sci8092Asthma/metabolism *Breath Tests Case-Control Studies Flow Injection Analysis/*methods Humans Hydrogen Peroxide/*analysis Reproducibility of Results Sensitivity and Specificity Spectrometry, Fluorescence/*methodsOct 5A method for the determination of hydrogen peroxide in exhaled breath condensate (EBC) by automated flow injection analysis (FIA) with fluorescence detection was developed and validated. In the enzymatic assay a fluorescent dimer of para-hydroxyphenyl acetic acid (HPAA) was formed by the redox coupling reaction between hydrogen peroxide and horseradish peroxidase (HRP). The calibration curve of hydrogen peroxide was linear over a range of 40-5000 nM. The coefficient of variation (CV) for within-day precision was 1-3%; for between-day precision, it was 2-5% over the validated range. The assay requires a small sample aliquot (150 microl) and no incubation time, and has an analytical runtime of < 2 min. It is therefore suitable for larger studies. The method was used to detect hydrogen peroxide in EBC of asthmatic patients and healthy volunteers. A statistically significant difference was found between patients with asthma (n = 19) and control subjects without asthma (n = 19), 780 nM versus 480 nM (P = 0.03).fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15315765 41570-0232 (Print) Journal Article Validation Studies15315765Occupational and Environmental Medicine, Sahlgrenska University Hospital and Academy at Goteborg University, Sankt Sigfridsgatan 85, SE-41266 Goteborg, Sweden. sophie.svensson@ymk.gu.se ~?gWLeung, T. F. Li, C. Y. Lam, C. W. Au, C. S. Yung, E. Chan, I. H. Wong, G. W. Fok, T. F.2004>The relation between obesity and asthmatic airway inflammation344-50Pediatr Allergy Immunol1547Adolescent Asthma/*complications/physiopathology Breath Tests Child Cross-Sectional Studies Female Humans Inflammation/*pathology/physiopathology Leukotriene B4/analysis Lung/*pathology/physiopathology Male Nitric Oxide/analysis Obesity/*complications Research Support, Non-U.S. Gov't Respiratory Function TestsAugEpidemiologic studies suggest increased asthma prevalence in obese subjects. However, the relation between obesity and airway inflammation remains unclear. This cross-sectional study aims to investigate the relation between obesity indices and exhaled nitric oxide (ENO) and leukotriene B(4) (LTB(4)) in children with asthma. Asthmatic patients aged 7-18 yr old were recruited. Weight-for-height Z score was calculated from anthropometry. ENO was measured by online single-breath method using a chemiluminescence analyzer, whereas LTB(4) concentrations in exhaled breath condensate (EBC) were quantified using competitive enzyme immunoassay. Ninety-two asthmatics and 23 controls were recruited. The mean ENO and LTB(4) concentrations in EBC were higher in asthmatic patients (87 p.p.b. and 40.5 pg/ml) than controls (25 p.p.b. and 18.7 pg/ml) (p < 0.0001 for both). Obesity, as defined by weight >120% median weight-for-height, was not associated with any alteration in ENO or LTB(4) concentrations in patients with asthma. Besides, these inflammatory markers did not differ between asthmatics in the highest and lowest quartiles of weight-for-height Z score. On multivariate analysis, ENO showed significant correlation with age (beta = 0.511, p < 0.0001), peripheral blood eosinophil count (beta = 0.222, p = 0.019), plasma total IgE concentration (beta = 0.187, p = 0.050) and forced expiratory volume in 1-s (FEV(1); beta = -0.221, p = 0.014). None of the factors was associated with LTB(4) concentration in EBC. In conclusion, ENO and LTB(4) concentration in EBC are increased in childhood asthma. However, these inflammatory markers did not differ between obese and non-obese children with asthma.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15305944 !0905-6157 (Print) Journal Article15305944Department of Paediatrics, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, Hong Kong. leung2142@cuhk.edu.hkU~?h=Rosias, P. Robroeks, C. Hendriks, J. Dompeling, E. Jobsis, Q.2004HExhaled breath condensate: a space odessey, where no one has gone before189-90; author reply 190 Eur Respir J241Biological Markers/*analysis Breath Tests Bronchoalveolar Lavage Fluid/chemistry Exhalation/physiology Female Humans Lung Diseases/*diagnosis Male Oxidative Stress/*physiology Sensitivity and Specificity Severity of Illness IndexJulfhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15293625 0903-1936 (Print) Comment Letter15293625t~?iGMcCafferty, J. B. Bradshaw, T. A. Tate, S. Greening, A. P. Innes, J. A.2004}Effects of breathing pattern and inspired air conditions on breath condensate volume, pH, nitrite, and protein concentrations694-8Thorax598Breath Tests Exhalation/*physiology Female Humans Hydrogen-Ion Concentration Inhalation/*physiology Male Nitrites/*analysis Prospective Studies Proteins/*analysis Research Support, Non-U.S. Gov't TemperatureAugBACKGROUND: The effects of breathing pattern and inspired air conditions on the volume and content of exhaled breath condensate (EBC) were investigated. METHODS: Total exhaled water (TEW), EBC volume, pH, nitrite and protein concentrations were measured in three groups of 10 healthy subjects breathing into a condenser at different target minute ventilations (Vm), tidal volumes (Vt), and inspired air conditions. RESULTS: The volumes of both TEW and EBC increased significantly with Vm. For Vm 7.5, 15 and 22.5 l/min, mean (SD) EBC was 627 (258) microl, 1019 (313) microl, and 1358 (364) microl, respectively (p<0.001) and TEW was 1879 (378) microl, 2986 (496) microl, and 4679 (700) microl, respectively (p<0.001). TEW was significantly higher than EBC, reflecting a condenser efficiency of 40% at a target Vm of 7.5 l/min which reduced to 29% at Vm 22.5 l/min. Lower Vt gave less TEW than higher Vt (26.6 v 30.7 microl/l, mean difference 4.1 (95% CI 2.6 to 5.6), p<0.001) and a smaller EBC volume (4.3 v 7.6 microl/l, mean difference 3.4 (95% CI 2.3 to 4.5), p<0.001). Cooler and drier inspired air yielded less water vapour and less breath condensate than standard conditions (p<0.05). Changes in the breathing pattern had no effect on EBC protein and nitrite concentrations and pH. CONCLUSION: These results show that condensate volume can be increased by using high Vt and increased Vm without compromising the dilution of the sample.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15282391 !0040-6376 (Print) Journal Article15282391gRespiratory Unit, Western General Hospital, Crewe Road, Edinburgh EH2 4XU, UK. john.mccafferty@ed.ac.uk ~?jZBack, E. I. Frindt, C. Nohr, D. Frank, J. Ziebach, R. Stern, M. Ranke, M. Biesalski, H. K.2004QAntioxidant deficiency in cystic fibrosis: when is the right time to take action?374-84Am J Clin Nutr802Adolescent Adult Aging/metabolism Antioxidants/*metabolism Ascorbic Acid Deficiency/blood Breath Tests Case-Control Studies Child Cholesterol/blood Cystic Fibrosis/blood/*metabolism Female Humans Isoprostanes/blood/deficiency Male Mouth Mucosa/metabolism *Oxidative Stress Research Support, Non-U.S. Gov't Thiobarbituric Acid Reactive Substances/metabolism Vitamin E Deficiency/bloodAugBACKGROUND: Little is known about age- and disease-related changes in prooxidant and antioxidant systems in patients with cystic fibrosis (CF). OBJECTIVE: We investigated changes in antioxidant concentrations and oxidative stress in plasma, buccal mucosal cells (BMCs), and breath condensate in patients with CF in relation to age and disease progression. DESIGN: We recruited 22 patients with CF as well as 35 healthy control subjects and conducted a cross-sectional study by dividing the participants into 4 age groups (<6 y, 6-11 y, 12-17 y, > or =18 y). We collected fasting blood samples, BMCs, and breath condensate. Carotenoids, alpha-tocopherol, vitamin C, protein carbonyls, thiobarbituric acid-reactive substances, and F(2)alpha-isoprostane were assessed. RESULTS: In patients with CF, plasma vitamin C concentrations, plasma and BMC alpha-tocopherol concentrations, and forced expiratory volume in 1 s (percentage predicted) decreased significantly with age. Plasma beta-carotene, beta-cryptoxanthin, and total lycopene were significantly lower in patients than in control subjects in all age groups. Furthermore, alpha-tocopherol and vitamin C plasma concentrations as well as alpha-tocopherol concentrations in BMCs were significantly lower in CF patients > or =18 y old, whereas all indicators of oxidative stress assessed were significantly higher than those same indicators in control subjects. CONCLUSIONS: Adult patients with CF in particular showed distinct vitamin deficits and elevated indicators of oxidative stress in plasma, BMCs, and breath condensate along with a progression of clinical status. We suggest that early in life dietary habits should be improved and that innovative supplementation strategies should be applied to optimize the antioxidant status of patients with CF.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15277158 !0002-9165 (Print) Journal Article15277158uInstitute of Biological Chemistry and Nutrition, University of Hohenheim, Garbenstrasse 30, 70593 Stuttgart, Germany. ~~?kHolz, O. Jorres, R. A.2004p[Non-invasive methods for monitoring airway inflammation: a comparison of expenditures, gain and clinical value]510-5 Pneumologie587Airway Obstruction/*diagnosis/*physiopathology Breath Tests English Abstract Humans Inflammation/diagnosis/physiopathology Monitoring, Physiologic/methods Nitric Oxide/analysis Sputum/chemistryJul{Among the noninvasive procedures for the assessment of airway inflammation, the analysis of spontaneous sputum is currently the only method, the expenses of which are covered by health insurance in Germany. It can easily be used for semiquantitive cytological analyses by practising pneumologists. Recent data also indicate the usefulness of sputum induction, particularly in asthma diagnosis and therapy control, and demonstrate its capability of reducing total costs per patient. In contrast to sputum analysis, the measurement of exhaled nitric oxide (NO) yields a read-out without time delay. NO as associated with eosinophils also seems suitable for monitoring airway inflammation. The number of studies regarding NO, both its pathophysiological role and clinical use, is far greater than that regarding any other marker of exhaled air. Measurements are easy and fast, but the costs of analysers are still prohibitive in clinical practice. The analysis of other compounds of exhaled air, particularly those of exhaled breath condensate (EBC), offers fascinating perspectives, owing to the scope of markers that might be measured, and could enable the assessment of multivariate profiles that are useful for diagnosis and therapy control. Currently, however, the method still faces methodological questions, and data indicating its usefulness and cost-efficiency in clinical practice are scarce. Compared to NO, the expenses per measurement in clinical use are mainly due to the costs per marker detection after sampling, as well as storage and transport of samples. The on-site analysis of pH in the EBC could be a first step to circumvent this obstacle.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15257474 (0934-8387 (Print) Journal Article Review15257474fNichtinvasive Verfahren zum Nachweis der Atemwegsentzundung im Vergleich Kosten - Nutzen - Wertigkeit.nKrankenhaus Grosshansdorf, Zentrum fur Pneumologie und Thoraxchirurgie, Grosshansdorf. o.holz@pulmoresearch.de~?l Effros, R. M.2004<Exhaled breath condensate acidification in acute lung injury682; author reply 683 Respir Med987Breath Tests/methods Humans Hydrogen-Ion Concentration Respiratory Distress Syndrome, Adult/*diagnosis Specimen Handling/methodsJulfhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15250237 0954-6111 (Print) Comment Letter15250237 G~?mOMajewska, E. Kasielski, M. Luczynski, R. Bartosz, G. Bialasiewicz, P. Nowak, D.2004Elevated exhalation of hydrogen peroxide and thiobarbituric acid reactive substances in patients with community acquired pneumonia669-76 Respir Med987SAdult Aged Biological Markers/analysis Breath Tests/methods C-Reactive Protein/metabolism Community-Acquired Infections/metabolism Female Humans Hydrogen Peroxide/*metabolism Male Middle Aged Oxidative Stress Pneumonia, Bacterial/drug therapy/*metabolism Research Support, Non-U.S. Gov't Thiobarbituric Acid Reactive Substances/*metabolismJulBACKGROUND: Bacterial pneumonia involves influx of activated phagocytes into distal airways. These cells release oxidants including H2O2, that may be exhaled or induce peroxidative damage to lung tissues with formation of thiobarbituric reactive substances (TBARs). STUDY OBJECTIVES: To determine whether concentrations of H2O2 and TBARs in exhaled breath condensate (EBC) is elevated and correlate with systemic response to pneumonia during 10 days of hospital treatment. DESIGN: The concentration of H2O2 and TBARs was measured in EBC of 43 inpatients with community acquired pneumonia (CAP) and 20 healthy never smoked subjects over 10 days and were accompanied by monitoring of WBC count, serum concentration of C-reactive protein (CRP) and peroxyl radical-trapping capacity. RESULTS: Patients with CAP exhaled 4.6-, 3.7-, 3.9-, 3.3-times more H2O2 than healthy controls at 1st, 3rd, 5th and 10th day of treatment (P<0.05), respectively. EBC concentrations of TBARs were elevated at 1st and 3rd day. H2O2 and TBARs levels decreased along with treatment course. Correlation (P<0.05) was found between H2O2 levels and CRP and WBC count (r = 0.31) at 1st day and between TBARs and CRP at 5th (r = 0.34) and 10th day (r = 0.46). The mean H2O2 exhalation estimated over ten days of treatment correlated with pneumonic chest X-ray score (r = 0.42), CRP levels (r = 0.46) and WBC count (r = 0.33) at admission (P<0.05). CONCLUSIONS: Pneumonia is accompanied by oxidative stress in airways that moderately correlates with intensity of systemic inflammatory response. Determination of H2O2 in EBC may be helpful for non-invasive monitoring of oxidants production during lower respiratory tract infection.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15250234 !0954-6111 (Print) Journal Article15250234Department of Experimental and Clinical Physiology, Institute of Physiology and Biochemistry, Medical University of Lodz, Mazowiecka St. 6/8, 92-215 Lodz, Poland. ~?nPBucchioni, E. Csoma, Z. Allegra, L. Chung, K. F. Barnes, P. J. Kharitonov, S. A.2004ZAdenosine 5'-monophosphate increases levels of leukotrienes in breath condensate in asthma651-5 Respir Med987Adenosine Monophosphate/*diagnostic use Adult Asthma/*diagnosis/physiopathology Biological Markers/analysis Breath Tests/methods Bronchial Hyperreactivity/diagnosis Bronchial Provocation Tests/methods Cysteine/*metabolism Female Forced Expiratory Volume Histamine Release/drug effects Humans Inflammation Mediators/metabolism Leukotrienes/*metabolism Male Methacholine Chloride/diagnostic use Vital CapacityJulHyperresponsiveness (AHR) is a key physiological abnormality in asthma. In clinical and research studies AHR is measured bronchial challenge, with methacholine (MCh), but more recently with adenosine-5'-monophosphate (AMP). In the search for markers of airway inflammation in asthmatic patients, we measured the concentrations of histamine and cysteinyl-leukotrienes (cys-LTs) before and after MCh and AMP challenges in the exhaled breath condensate of 13 patients with mild asthma (FEV1 78.5%pred) and nine healthy non-smokers, using specific enzyme immunoassays. With methacholine challenge we did not find any differences between asthmatics and normal subjects in the pre- and post-challenge concentrations of cys-LTs: 27.2+/-1.4 vs. 29.2+/-1.2 pg/ml and 26.3+/-2.2 vs. 27.5+/-4.2 pg/ml, respectively or histamine: 5.1+/-0.4 vs. 5.1+/-0.6 nM and 4.5+/-0.4 vs. 4.4+/-0.3 nM; P>0.05). In asthmatic patients cys-LT levels were significantly higher after AMP challenge (56.2+/-9.7 vs. 31.7+/-6.9 pg/ml; P<0.05); but there was no difference in healthy subjects (27.2+/-4.6 vs. 30.3+/-4.7 pg/ml). There was no difference in histamine concentrations in asthmatic (5.9+/-1.8 vs. 4.5+/-0.5 nM), or healthy subjects (5.5+/-0.4 vs.5.7+/-0.9 nM) after AMP challenge. In conclusion, our results show that the cys-LTs are increased in exhaled breath condensate after AMP challenge, which may indicate that the AMP acts indirectly by releasing cys-LTs from primed mast cells. The detection of LTs and histamine in exhaled breath condensate may be useful in monitoring asthma.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15250231 40954-6111 (Print) Evaluation Studies Journal Article15250231{Department of Thoracic Medicine, National Heart and Lung Institute, Imperial College, Dovehouse Street, London SW3 6LY, UK.~?oRKhasina, M. A. Dvinskaia, S. A. Beloglazova, S. I. Khasina, MIu Puchinskaia, T. F.2004[Vapor condensate of exhaled air in evaluating the impaired metabolism of the bronchopulmanory system in nonspecific lung diseases]15-7Klin Lab Diagn5Adolescent Adult Breath Tests/methods Bronchitis, Chronic/drug therapy/enzymology/*metabolism English Abstract Humans Male Pneumonia, Bacterial/drug therapy/enzymology/*metabolismMayThe degree of metabolic rehabilitation of the bronchopulmonary system was evaluated in non-specific pulmonary diseases, like pneumonia or chronic obstructive bronchitis, by using the data of biochemical testing of the exhaled-air vapor condensate. Nine parameters were investigated, i.e. enzymes alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase, alkaline phosphatase (AP), gamma-glutamate amino-transpeptidase (GGT) as well as parameters of protein metabolism--common protein, seromucoid (SC), C-reactive protein and urea. AST, ALT, AP, GGT, SC and urea were acknowledged as the most informative parameters. The results are indicative of that the recovery of metabolic processes in the bronchopulmonary system was not completed.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15230110 !0869-2084 (Print) Journal Article15230110Kondensat parov vydykhaemogo vozdukha v otsenke stepeni narusheniia metabolizma bronkholegochnoi sistemy pri nespetsificheskikh zabolevaniiakh legkikh. ~?p8Niimi, A. Nguyen, L. T. Usmani, O. Mann, B. Chung, K. F.2004ZReduced pH and chloride levels in exhaled breath condensate of patients with chronic cough608-12Thorax597;Adult Asthma/complications Breath Tests/methods Bronchiectasis/complications Capsaicin/pharmacology Chlorides/*metabolism Chronic Disease Cough/*etiology/metabolism Female Gastroesophageal Reflux/complications Humans Hydrogen-Ion Concentration Male Research Support, Non-U.S. Gov't Rhinitis/complications StatisticsJulBACKGROUND: Increased hydrogen and reduced chloride ionic environments of the airways are conducive to the stimulation of cough. However, the constituents of the local milieu of the airways of patients with chronic cough are unknown. METHODS: The pH and chloride levels in exhaled breath condensate and capsaicin cough threshold (C5) were measured in 50 patients with chronic cough and in 16 healthy controls. pH and chloride measurements were repeated after capsaicin challenge in those with cough. The cause of cough was asthma (n = 13), postnasal drip/rhinitis (n = 7), gastro-oesophageal reflux (n = 5), bronchiectasis (n = 5), but remained unidentified in 20. RESULTS: Compared with controls, patients with chronic cough had lower pH (mean 7.9 v 8.3, 95% CI of difference -0.5 to -0.2, p<0.0001), chloride levels (median 4 v 6 mmol/l, 95% CI -3.1 to -0.2, p = 0.007), and C5 (median 3.9 v 125 micro M, 95% CI -270.0 to -17.6, p = 0.002). The pH levels were different in the six subgroups including controls, and were reduced in all diagnostic subgroups of patients with cough compared with controls but did not differ between them. Chloride levels were significantly different in the six subgroups but were lower than controls in only the gastro-oesophageal reflux subgroup. There was a weak but significant correlation between chloride levels and C5 when all participants were analysed together, but not between pH and C5 or chloride levels. pH and chloride levels did not change after capsaicin challenge. CONCLUSIONS: The epithelial lining fluid of patients with chronic cough has a reduced pH and reduced chloride levels which could contribute to the enhanced cough reflex.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15223872 J0040-6376 (Print) Clinical Trial Controlled Clinical Trial Journal Article15223872Department of Thoracic Medicine, National Heart and Lung Institute, Imperial College and Royal Brompton Hospital, Dovehouse Street, London SW3 6LY, UK.~?q Effros, R. M.2004Exhaled breath condensate pH961-2; author reply 962 Eur Respir J236Acid-Base Equilibrium/physiology Biological Markers Breath Tests/*methods Extravascular Lung Water/*chemistry Humans Hydrogen-Ion Concentration Lung Diseases/*diagnosis/physiopathology Respiratory System/*physiopathologyJunfhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15219015 0903-1936 (Print) Comment Letter15219015 ~?rRCarpagnano, G. E. Foschino-Barbaro, M. P. Resta, O. Gramiccioni, E. Carpagnano, F.2004^Endothelin-1 is increased in the breath condensate of patients with non-small-cell lung cancer180-4Oncology663Aged Breath Tests Carcinoma, Non-Small-Cell Lung/*diagnosis/*metabolism Early Diagnosis Endothelin-1/*metabolism Female Humans Lung Neoplasms/*diagnosis/*metabolism Male Middle Aged Predictive Value of Tests Reproducibility of Results Tumor Markers, Biological/*metabolismOne recent line of cancer research is currently directed to the study of growth factors. Of increasing interest is endothelin-1 (ET-1), a mitogenic factor already investigated in several human cancer cell lines, which has been found to participate in the development and progression of tumours. This peptide has an important role also in non-small-cell lung cancer (NSCLC) where ET-1 expression has been found in 100% of cell lines. OBJECTIVES: The aim of this study was to measure ET-1 concentrations in the airways of patients with NSCLC using a completely non-invasive procedure--the breath condensate--and to verify the involvement of this peptide in the growth of lung tumours. METHODS: We enrolled 30 patients (17 men, median age 63 years; range 53-74) with histological evidence of NSCLC and 15 healthy controls (9 men, median age 59 years; range 52-70). ET-1 was measured in the exhaled breath condensate by means of a specific enzyme immunoassay kit. RESULTS: Higher concentrations of exhaled ET-1 were found in NSCLC patients (8.3 +/- 0.7 pg/ml) compared to controls (5.2 +/- 0.5 pg/ml, p < 0.0001). A statistically significant difference was observed between patients with distant metastases (stage IV) of NSCLC (8.9 +/- 0.6 pg/ml) and those with locoregional disease (stage I-III) (7.9 +/- 0.5 pg/ml). A significant reduction in ET-1 levels was found in 14 patients after surgical removal of the tumour either associated with or without adjuvant chemotherapy (6.3 +/- 0.5 vs. 7.9 +/- 0.4 pg/ml, p < 0.0001). CONCLUSIONS: These findings suggest that the measurement of ET-1 in the breath condensate of patients with NSCLC could be proposed as a marker for early detection of NSCLC as well as for monitoring reduction or progression of the neoplasm in the follow-up of treated patients.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15218307 !0030-2414 (Print) Journal Article15218307BInstitute of Respiratory Disease, University of Bari, Bari, Italy. ~?shRosias, P. P. Dompeling, E. Dentener, M. A. Pennings, H. J. Hendriks, H. J. Van Iersel, M. P. Jobsis, Q.2004gChildhood asthma: exhaled markers of airway inflammation, asthma control score, and lung function tests107-14Pediatr Pulmonol382/Adolescent Albumins/analysis Asthma/*diagnosis/immunology *Breath Tests C-Reactive Protein/analysis Carbon Monoxide/analysis Case-Control Studies Child Comparative Study Cytokines/analysis Enzyme-Linked Immunosorbent Assay Exhalation Humans Hydrogen-Ion Concentration Inflammation Mediators/*analysis Intercellular Adhesion Molecule-1/analysis Interleukin-6/analysis Interleukin-8/analysis Nitric Oxide/analysis Patient Selection Questionnaires Receptors, Tumor Necrosis Factor, Type II/analysis Respiratory Function Tests Tumor Necrosis Factor-alpha/analysisAuggExhaled markers of airway inflammation become increasingly important in the management of childhood asthma. The aims of the present study are: 1) to compare exhaled markers of inflammation (nitric oxide, carbon monoxide, and acidity of breath condensate) with conventional asthma measures (lung function tests and asthma control score) in childhood asthma; and 2) to investigate the detectability of albumin, CRP, IL-6, IL-8, TNF-alpha, sICAM-1, and sTNF-R75 in the exhaled breath condensate (EBC) of asthmatic children. Thirty-two children with mild to moderate persistent asthma and healthy controls aged 6-12 years were studied. We measured exhaled NO and CO, and subsequently EBC was collected. Inflammatory mediators in EBC were measured using an enzyme-linked immunosorbent assay. Respiratory symptoms and asthma control were assessed using the asthma control questionnaire (ACQ) of Juniper et al. (Eur Respir J 1999;14:902-907). Exhaled NO showed a significant correlation with exhaled CO (r = 0.59, P < 0.05) and FEV1 (r = -0.59, P < 0.05), but not with ACQ score (r = 0.48, P = 0.06). Exhaled CO was correlated with prebronchodilator FEV1 (r = -0.45, P < 0.05), but not with asthma control (r = 0.18, P = 0.35). Acidity of EBC was significantly lower in asthmatic children than in healthy controls (P < 0.05), but did not correlate with any of the conventional asthma measures. We were not able to demonstrate the presence of CRP, IL-6, IL-8, TNF-alpha, sICAM-1, and sTNF-R75 in EBC. Albumin was found in two EBC samples of asthmatic children. We conclude that exhaled NO had a better correlation with lung function parameters and asthma control than exhaled CO and acidity of EBC, in mild to moderate persistent childhood asthma. However, exhaled NO, CO, and deaerated pH of EBC did not differ between asthmatic children and controls, possibly because of a too homogeneous and well-controlled study population. To further evaluate the clinical utility of exhaled markers in monitoring childhood asthma, more studies are required on a wider range of asthma severity, and preferably with repeated measurements of markers and of asthma control.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15211692 !8755-6863 (Print) Journal Article15211692nDepartment of Pediatric Pulmonology, University Hospital Maastricht, The Netherlands. p.rosias@orbisconcern.nl&~?tlGianazza, E. Allegra, L. Bucchioni, E. Eberini, I. Puglisi, L. Blasi, F. Terzano, C. Wait, R. Sirtori, C. R.2004tIncreased keratin content detected by proteomic analysis of exhaled breath condensate from healthy persons who smoke51-4Am J Med1171Aged *Breath Tests Electrophoresis, Gel, Two-Dimensional Electrophoresis, Polyacrylamide Gel Female Health Status Humans Keratin/*analysis Male Middle Aged Proteomics/instrumentation/*methods *Smoking/metabolismJul 1fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15210388 !0002-9343 (Print) Journal Article15210388Dipartimento di Scienze Farmacologiche, Gruppo di Studio per la Proteomica e la Struttura delle Proteine, Universita degli Studi di Milano, Milan, Italy. ~?uQCarpagnano, G. E. Barnes, P. J. Francis, J. Wilson, N. Bush, A. Kharitonov, S. A.2004rBreath condensate pH in children with cystic fibrosis and asthma: a new noninvasive marker of airway inflammation?2005-10Chest1256Asthma/*diagnosis *Breath Tests Case-Control Studies Child Child, Preschool Comparative Study Cystic Fibrosis/*diagnosis F2-Isoprostanes/analysis Female Humans *Hydrogen-Ion Concentration Inflammation Mediators/*analysis Leukotriene B4/analysis Male Predictive Value of Tests Probability Prognosis Prospective Studies Reference Values Respiratory Function Tests Risk Assessment Sensitivity and Specificity Severity of Illness Index Statistics, NonparametricJunSTUDY OBJECTIVES: The noninvasive assessment and monitoring of airway inflammation could be important in respiratory disease. The pH of exhaled breath condensate (EBC) is a promising marker. Although pH has been measured in the EBC of adults with inflammatory airway diseases, no study has measured this in children. DESIGN: This study aimed to assess whether there is a change in pH in the EBC of children with cystic fibrosis (CF) and asthma, and to try to determine whether pH could be used as a marker of airway inflammation. Furthermore, the relationships among EBC pH, severity of disease, and oxidative stress were studied. PATIENTS AND METHODS: We studied 20 children with CF (mean [+/- SEM] age, 7 +/- 3 years), 20 children with asthma (mean age, 7 +/- 2 years), and 15 age-matched healthy children (mean age, 7 +/- 2 years). The pH of EBC was measured using a pH meter. MEASUREMENTS AND RESULTS: Lower pH values were observed in the EBC of children with CF and asthma compared to control subjects (mean pH, 7.23 +/- 0.03 and 7.42 +/- 0.01 vs 7.85 +/- 0.02, respectively). Furthermore, relationships among EBC pH, severity of asthma, and the presence of an infective exacerbation of CF was found. There was a negative correlation between exhaled pH and exhaled leukotriene B(4) concentrations (r = -0.5; p < 0.005). CONCLUSION: We conclude that the measurement of EBC pH may be useful in the evaluation of airway inflammation in children with asthma and CF.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15189915 !0012-3692 (Print) Journal Article15189915_Department of Thoracic Medicine, National Heart & Lung Institute, Imperial College, London, UK. ~?vLCap, P. Chladek, J. Pehal, F. Maly, M. Petru, V. Barnes, P. J. Montuschi, P.2004[Gas chromatography/mass spectrometry analysis of exhaled leukotrienes in asthmatic patients465-70Thorax596QAdministration, Inhalation Adrenal Cortex Hormones/*administration & dosage Adult Asthma/drug therapy/*metabolism Breath Tests Child Chromatography, Gas Cross-Sectional Studies Female Forced Expiratory Volume/physiology Humans Leukotrienes/*analysis Male Research Support, Non-U.S. Gov't Spectrum Analysis, Mass Vital Capacity/physiologyJungBACKGROUND: Leukotriene-like immunoreactivity has been detected in exhaled breath condensate (EBC), but definitive evidence for the presence of leukotrienes (LTs) in this biological fluid is not available. A study was undertaken to determine whether LTC(4), LTD(4), LTE(4), and LTB(4) are measurable in EBC by gas chromatography/mass spectrometry and to quantify exhaled LTs in adults and children with asthma and in control subjects. METHODS: Twenty eight adults and 33 children with mild to moderate persistent asthma treated with inhaled corticosteroids and age matched healthy controls (50 adults and 50 children) were studied. LTB(4), LTC(4), LTD(4), and LTE(4) in EBC were measured by gas chromatography/mass spectrometry. RESULTS: LTD(4), LTE(4), and LTB(4) were detectable in all samples. Concentrations of LTC(4) in EBC were either close to or below the detection limit of 1 pg/ml. Median exhaled LTD(4), LTE(4), and LTB(4) concentrations in asthmatic adults were increased 4.1-fold (p<0.001), 1.8-fold (p<0.01), and 2.6-fold (p<0.001), respectively, compared with values in healthy adults. Median exhaled LTD(4), LTE(4), and LTB(4) concentrations in asthmatic children were increased 2.8-fold (p<0.001), 1.3-fold (p<0.001), and 1.6-fold (p<0.001), respectively, compared with those in healthy children. In patients with asthma there was a correlation between exhaled LTD(4) and LTE(4) in both adults (r = 0.87, p<0.0001) and children (r = 0.78, p<0.0001). CONCLUSIONS: Gas chromatography/mass spectrometry can be used to accurately quantify exhaled LTs which are increased in asthmatic adults and children compared with controls.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15170025 !0040-6376 (Print) Journal Article15170025_Department of Allergology and Clinical Immunology, Hospital Na Homolce, Prague, Czech Republic. I~?w?van Beurden, W. J. Smeenk, F. W. Harff, G. A. Dekhuijzen, P. N.2003gMarkers of inflammation and oxidative stress during lower respiratory tract infections in COPD patients273-80Monaldi Arch Chest Dis5946Administration, Inhalation Adrenal Cortex Hormones/therapeutic use Aged Aged, 80 and over Analysis of Variance Blood Chemical Analysis Comparative Study Disease Progression Female Hospitalization Humans Inflammation Mediators/*analysis Male Middle Aged Oxidative Stress/*physiology Pneumonia/complications/*diagnosis Probability Prognosis Pulmonary Disease, Chronic Obstructive/complications/*diagnosis/drug therapy Respiratory Function Tests Risk Assessment Sampling Studies Sensitivity and Specificity Severity of Illness Index Spirometry Statistics, NonparametricOct-DecBACKGROUND: Lower respiratory tract infections (LRTI) occur frequently in patients with Chronic Obstructive Pulmonary Disease (COPD), and are a major cause of morbidity, mortality and health care utilization. The aim of this study was to investigate if non- or less invasive markers of inflammation and oxidative stress can predict the course of the infections. METHODS: Twenty-five COPD patients who were admitted to hospital with a LRTI were included. Within 24 hours after admittance, spirometry (FEV1, FVC, MEF50), measurement of hydrogen peroxide (H2O2) in exhaled breath condensate (EBC), symptom scores and analyses of ESR, CRP, ECP, and MPO in serum were performed. All patients were treated with intravenous dexamethasone, nebulised salbutamol/ipratropium and, if needed, antibiotics. The tests were repeated at day 2, 3, 7 and 30. RESULTS: Complete data of the first four visits were collected in 19 patients. The H2O2 concentration and spirometry parameters did not change significantly during the study period. CRP, ESR and MPO levels decreased significantly during treatment, while the other serum inflammatory parameters did not change. There were no significant correlations between H2O2 concentration, spirometry and serum inflammatory parameters. CONCLUSIONS: In conclusion, this study showed no significant changes in H2O2 concentration in EBC, or spirometry during treatment of a LRTI in COPD patients. In contrast, several serum inflammatory markers did decrease during hospitalization, thus providing a simple tool to monitor exacerbations.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15148836 !1122-0643 (Print) Journal Article15148836fDepartment of Pulmonology, Catharina Hospital, Eindhoven, The Netherlands. wendy.beurdenvan@wanadoo.nl ~?x8Leung, T. F. Wong, G. W. Ko, F. W. Lam, C. W. Fok, T. F.2004hIncreased macrophage-derived chemokine in exhaled breath condensate and plasma from children with asthma786-91Clin Exp Allergy345mAdolescent Analysis of Variance Asthma/drug therapy/*immunology Biological Markers/analysis/blood Breath Tests Case-Control Studies Chemokines/*analysis/blood Chemokines, CC/analysis/blood Child Chronic Disease Female Glucocorticoids/therapeutic use Humans Macrophages/*immunology Male Research Support, Non-U.S. Gov't Statistics, Nonparametric Th2 Cells/immunologyMayBACKGROUND: Type 2 helper T lymphocyte-specific chemokines including macrophage-derived chemokine (MDC), thymus and activation-regulated chemokine (TARC) and eotaxin are important mediators for allergic airway inflammation. OBJECTIVE: We investigated whether these chemokines can be detected in exhaled breath condensate (EBC) and their relation to childhood asthma. METHODS: Asthmatics recruited from paediatric clinics of a university teaching hospital were classified into intermittent asthma (IA) and persistent asthma (PA) according to Global Initiative for Asthma guidelines. EBC was collected by a disposable collection kit, whereas fractional exhaled nitric oxide (FENO) was measured by a chemiluminescence analyser. Concentrations of MDC, TARC and eotaxin in both EBC and plasma were measured using sandwich enzyme immunoassay. The intra-subject reproducibility of exhaled chemokine measurements was determined by co-efficients of variation (CV). RESULTS: Forty-eight patients with PA, 36 children with IA and 18 controls were recruited. MDC and eotaxin were present in EBC from nearly all subjects, whereas TARC could be measured in EBC from 33 (32%) subjects only. The median MDC concentration in EBC was higher in PA (117 pg/mL) as compared with IA (106 pg/mL) and controls (105 pg/mL; P=0.003 for both). The median plasma MDC concentration in PA (648 pg/mL) was also higher than that in IA (520 pg/mL; P=0.002) and controls (490 pg/mL; P=0.008). The median plasma TARC concentration was also increased in PA as compared with IA (72 pg/mL vs. 35 pg/mL; P=0.004). MDC concentrations in EBC were lower in patients with PA who received high-dose inhaled corticosteroid (P=0.005). FENO was significantly higher in asthmatics than controls (P<0.0001), but it was not associated with chemokines in EBC or plasma. The mean (range) CV for measuring MDC, TARC and eotaxin in EBC (n=6) were 5.5 (2.0-7.2%), 8.8 (3.6-14.4%) and 5.2 (2.8-7.9%), respectively. CONCLUSIONS: Our results suggest that MDC in EBC and MDC and TARC in plasma are increased in children with PA as compared with IA or control. MDC concentrations in EBC are suppressed in patients on high-dose inhaled corticosteroid treatment.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15144472 !0954-7894 (Print) Journal Article15144472lDepartments of Paediatrics, The Chinese University of Hong Kong, Hong Kong SAR, China. leung2142@cuhk.edu.hk~?yoWeissmann, N. Vogels, H. Schermuly, R. T. Ghofrani, H. A. Hanze, J. Fink, L. Rose, F. Seeger, W. Grimminger, F.2004:Measurement of exhaled hydrogen peroxide from rabbit lungs259-64 Biol Chem3853-4Animals Comparative Study Exhalation/*physiology Female Hydrogen Peroxide/analysis/*metabolism Intubation, Intratracheal/methods Lung/chemistry/*metabolism Male Pulmonary Ventilation/physiology Rabbits Research Support, Non-U.S. Gov'tMar-AprExhaled H2O2 is considered an indicator of lung inflammatory and oxidative stress. Moreover, H2O2 may be involved in signal transduction processes. It is not fully elucidated to what extent (i) H2O2 escapes from the intravascular compartment, and (ii) pulmonary H2O2 generation and nasopharyngeal H2O2 generation contribute to exhaled H2O2. We investigated H2O2 concentrations in breath condensate from isolated buffer-perfused and ventilated rabbit lungs, and from both intubated and spontaneously breathing rabbits with a horseradish peroxidase/2',7'dichlorofluorescin assay. For the perfused lungs, a H2O2 concentration of 58 +/- 19 nM was found. Addition of H2O2 to the buffer fluid resulted in only minute appearance in the exhaled air (<0.001%). Levels of exhaled H2O2 in intubated rabbits and perfused lungs were virtually identical. Nearly ten-fold higher levels were detected in spontaneously breathing rabbits. Decreasing the inspired oxygen concentration from 21% to 1% resulted in a tendency toward decreased H2O2 exhalation in perfused lungs. In contrast, phorbol-12-myristate-13-acetate (PMA) prompted a approximately 4-fold increase in H2O2 exhalation. We conclude that the horseradish peroxidase/2',7'dichlorofluorescin assay is a feasible technique to measure H2O2 in exhaled breath condensate in rabbits. When collecting exhaled air via the tracheal tube, the signal represents pulmonary H2O2 generation with the contribution of the remaining body being negligible.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15134339 !1431-6730 (Print) Journal Article15134339Department of Internal Medicine, Justus-Liebig-University Giessen, Klinikstrasse 36, D-35392 Giessen, Germany. Norbert.Weissmann@innere.med.uni-giessen.de 2~?zmSzkudlarek, U. Zdziechowski, A. Witkowski, K. Kasielski, M. Luczynska, M. Luczynski, R. Sarniak, A. Nowak, D.2004VEffect of inhaled N-acetylcysteine on hydrogen peroxide exhalation in healthy subjects155-62Pulm Pharmacol Ther173Acetylcysteine/administration & dosage/*pharmacology Administration, Inhalation Adult Antioxidants/administration & dosage/*pharmacology Breath Tests Exhalation Female Humans Hydrogen Peroxide/*metabolism Male Research Support, Non-U.S. Gov'tN-acetylcysteine (NAC) has antioxidant properties and its oral administration decreased H(2)O(2) exhalation in patients with chronic obstructive pulmonary disease. In this study we tested whether inhaled NAC could suppress H(2)O(2) levels in exhaled breath condensate (EBC) of eight healthy subjects that have never smoked (never-smokers). Original NAC solution (ACC vial, 300 mg NAC in 3 ml solvent), NAC-placebo (vehicle), sterile 0.9% NaCl or distilled water were nebulized via the pneumatic De Vilbiss nebulizer once daily every 7 days and H(2)O(2) and thiols exhalation was measured just before, 30 min and 3 h after the end of drug administration. Additional in vitro experiments were performed to evaluate NAC stability during nebulization, reactivity with H(2)O(2) and possible H(2)O(2) generation in aqueous NAC solutions. NAC almost completely abolished H(2)O(2) exhalation 30 min after inhalation (0.02+/-0.04 vs. 0.21+/-0.09 microM, p<0.001). However, 3 h later the H(2)O(2) levels raised 1.8-fold from baseline (p<0.01). Other inhaled solutions did not affect H(2)O(2) levels. Mean thiol concentration in EBC rose (p<0.05) after treatment with NAC and reached 1.03+/-0.48 microM at 3 h. Although, 25 and 50 mM NAC completely inhibited H(2)O(2)-peroxidase-luminol-dependent chemiluminescence, detectable amounts of H(2)O(2) were generated in NAC solutions. It was accompanied by moderate loss of -SH groups. Catalase and ascorbic acid prevented H(2)O(2) formation in NAC solutions. In conclusion inhaled NAC revealed biphasic effect on H(2)O(2) exhalation in healthy subjects, which depends on direct H(2)O(2) scavenging and H(2)O(2) generation related to drug oxidation. The net result of these processes may determine anti- or pro-oxidant action of inhaled NAC.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15123225 01094-5539 (Print) Clinical Trial Journal Article15123225Department of Experimental and Clinical Physiology, Institute of Physiology and Biochemistry, Medical University of Lodz, Mazowiecka 6/8, 92-215 Lodz, Poland.~?{Kharitonov, S. A.2004LExhaled markers of inflammatory lung diseases: ready for routine monitoring?175-92Swiss Med Wkly13413-14Alkanes/*metabolism Biological Markers Breath Tests Chronic Disease Ciliary Motility Disorders/metabolism Cough/metabolism Exhalation Humans Lung Diseases, Obstructive/*metabolism Nitric Oxide/*metabolism Reproducibility of Results Rhinitis/metabolismApr 3Assessing airway inflammation is important for investigating the underlying mechanisms of many lung diseases, including asthma and chronic obstructive pulmonary disease (COPD). Yet these are not measured directly in routine clinical practice because of the difficulties in monitoring inflammation. The presence and type of airway inflammation can be difficult to detect clinically, and may result in delays in initiating appropriate therapy. Non-invasive monitoring may assist in differential diagnosis of lung diseases, assessment of their severity and response to treatment. There is increasing evidence that breath analysis may have an important place in the diagnosis and clinical management of asthma, COPD, primary ciliary dyskinesia (PCD) and other major lung disease. The article reviews whether current noninvasive measurements of exhaled gases, such as nitric oxide (NO), hydrocarbons, inflammatory markers exhaled breath condensate (EBC) are ready for routine use in clinical practice.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15106031 (1424-7860 (Print) Journal Article Review15106031uNational Heart and Lung Institute, Imperial College, Royal Brompton Hospital, London, UK. s.kharitonov@imperial.ac.uk ~?|Deaton, C. M. Marlin, D. J. Smith, N. C. Smith, K. C. Newton, R. J. Gower, S. M. Cade, S. M. Roberts, C. A. Harris, P. A. Schroter, R. C. Kelly, F. J.2004Breath condensate hydrogen peroxide correlates with both airway cytology and epithelial lining fluid ascorbic acid concentration in the horse201-8Free Radic Res382Airway Obstruction/*diagnosis/pathology Animals Ascorbic Acid/*analysis/metabolism Breath Tests/*methods Bronchoalveolar Lavage Fluid/cytology Bronchoscopy Cell Count Comparative Study Dehydroascorbic Acid/analysis/metabolism Diagnosis, Differential *Disease Models, Animal Glutathione/analysis/metabolism *Horses Hydrogen Peroxide/*analysis/metabolism Inflammation/diagnosis/metabolism Mucus/cytology Neutrophils/cytology Respiratory Mucosa/chemistry Respiratory System/pathology SpectrophotometryFeb$The relationship between hydrogen peroxide (H2O2) concentration in expired breath condensate (EBC) and cytology of the respiratory tract obtained from tracheal wash (TW) or bronchoalveolar lavage (BAL), and epithelial lining fluid (ELF) antioxidant status is unknown. To examine this we analysed the concentration of H2O2 in breath condensate from healthy horses and horses affected by recurrent airway obstruction (RAO), a condition considered to be an animal model of human asthma. The degree of airway inflammation was determined by assessing TW inflammation as mucus, cell density and neutrophil scores, and by BAL cytology. ELF antioxidant status was determined by measurement of ascorbic acid, dehydroascorbate, reduced and oxidised glutathione, uric acid and alpha-tocopherol concentrations. RAO-affected horses with marked airway inflammation had significantly higher concentrations of breath condensate H2O2 than control horses and RAO-affected horses in the absence of inflammation (2.0 +/- 0.5 micromol/l. 0.4 +/- 0.2 micromol/l and 0.9 +/- 0.2 micromol/l H2O2, respectively; p < 0.0001). The concentration of breath condensate H2O2 was related inversely to the concentration of ascorbic acid in ELF (r = -0.80; p < 0.0001) and correlated positively with TW inflammation score (r = 0.76, p < 0.0001) and BAL neutrophil count (r = 0.80, p < 0.0001). We conclude that the concentration of H2O2 in breath condensate influences the ELF ascorbic acid concentration and provides a non-invasive diagnostic indicator of the severity of neutrophilic airway inflammation.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15104214 41071-5762 (Print) Evaluation Studies Journal Article15104214uCentre for Equine Studies, Animal Health Trust, Lanwades Park, Kentford, Suffolk CB8 7UU, UK. chris.deaton@aht.org.uk~?}0Gessner, C. Hammerschmidt, S. Kuhn, H. Wirtz, H.2004>[Expired diagnosis?--the potential of exhaled breath analysis]230-7 Pneumologie584*Breath Tests Cystic Fibrosis/diagnosis Diagnostic Tests, Routine/*methods English Abstract Humans Pulmonary Disease, Chronic Obstructive/diagnosisAprgAnalysis of breath condensate is an innovative approach to biochemical information from the lung. It provides a new tool to estimate and characterize the burden of oxidative and inflammatory processes in the airways/lung. Clinical applications in asthma, COPD and CF can be envisioned as well as determining organ-specific inflammation in mechanically ventilated patients or monitoring patients with transplanted lungs. However, besides inflammation other important areas have begun to be evaluated, such as the demonstration of p53 mutations in NSCLC patients or an increased ratio of EBC nitrite to tidal volume in mechanically stressed lungs. Of course a careful validation of each and every parameter is paramount to the use in clinical applications. The comparison to BAL is oftentimes called for but at the same time is not the comparison to a gold standard because of the well-known problems of BAL. The scope of this review is a summary of facts and theories concerning exhaled breath condensate generation, collection and analysis but at the same time the representation of the many aspects that remain to be resolved.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15098160 (0934-8387 (Print) Journal Article Review15098160CGehauchte Diagnosen? Zum Potenzial von Atemkondensatuntersuchungen.Medizinische Klinik und Poliklinik I, Universitat Leipzig, Johannisallee 32, 04103 Leipzig, Germany. gesc@medizin.uni-leipzig.de&~?~Wyse, C. A. Preston, T. Yam, P. S. Sutton, D. G. Christley, R. M. Hotchkiss, J. W. Mills, C. A. Glidle, A. Cumming, D. R. Cooper, J. M. Love, S.2004?Current and future uses of breath analysis as a diagnostic tool353-60Vet Rec15412Animals Breath Tests/instrumentation/*methods Gastrointestinal Diseases/diagnosis/*veterinary Research Support, Non-U.S. Gov't Veterinary Medicine/*trendsMar 20pThe analysis of exhaled breath is a potentially useful method for application in veterinary diagnostics. Breath samples can be easily collected from animals by means of a face mask or collection chamber with minimal disturbance to the animal. After the administration of a 13C-labelled compound the recovery of 13C in breath can be used to investigate gastrointestinal and digestive functions. Exhaled hydrogen can be used to assess orocaecal transit time and malabsorption, and exhaled nitric oxide, carbon monoxide and pentane can be used to assess oxidative stress and inflammation. The analysis of compounds dissolved in the aqueous phase of breath (the exhaled breath condensate) can be used to assess airway inflammation. This review summarises the current status of breath analysis in veterinary medicine, and analyses its potential for assessing animal health and disease.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15074325 (0042-4900 (Print) Journal Article Review15074325fInstitute of Comparative Medicine, University of Glasgow Veterinary School, Bearsden, Glasgow G61 1QH.~? Griese, M.2004Exhaled breath condensate14-5Pediatr Pulmonol Suppl26*Breath Tests Humans Leukotriene B4/analysis Lung Diseases/diagnosis Monitoring, Physiologic/*methods Reproducibility of Results Respiratory Function Testsfhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15029580 !1054-187X (Print) Journal Article15029580TChildren's Hospital University of Munich, Germany. mgriese@helios.med.unimuenchen.de ~?_Psathakis, K. Papatheodorou, G. Plataki, M. Panagou, P. Loukides, S. Siafakas, N. M. Bouros, D.20048-Isoprostane, a marker of oxidative stress, is increased in the expired breath condensate of patients with pulmonary sarcoidosis1005-11Chest1253rAdult Biological Markers/analysis *Breath Tests Calcium/blood Cross-Sectional Studies *Dinoprost/*analogs & derivatives F2-Isoprostanes/*metabolism Female Forced Expiratory Volume Humans Lung Volume Measurements Male Middle Aged *Oxidative Stress Peptidyl-Dipeptidase A/blood Pulmonary Diffusing Capacity Sarcoidosis, Pulmonary/*metabolism/physiopathology Vital CapacityMarfSTUDY OBJECTIVE: 8-Isoprostane is considered an index of oxidative stress. Measurement of 8-isoprostane in the expired breath condensate, a totally noninvasive method, has not been used to explore the level of inflammation in pulmonary sarcoidosis. Therefore, the aim of our study was to measure the levels of 8-isoprostane in the expired breath condensate of patients with sarcoidosis, and to investigate the relation of 8-isoprostane level to disease activity. PATIENTS: We investigated 30 patients with pulmonary sarcoidosis (active disease, n = 14; nonactive disease, n = 16) and 12 healthy subjects as control group. METHODS: 8-Isoprostane was measured in the expired breath condensate of all subjects, and its levels were compared between the control and sarcoidosis groups as well as between the subgroups of patients with active and nonactive disease. In the group with sarcoidosis, 8-isoprostane levels were further correlated with markers that may reflect disease activity, such as serum angiotensin-converting enzyme (sACE) level, serum calcium level, and pulmonary function test results. RESULTS: The concentration of 8-isoprostane was increased in patients with sarcoidosis compared to control subjects (mean, 64.23 pg/mL; 95% confidence interval [CI], 37.00 to 91.46 pg/mL; vs mean, 20.75 pg/mL; 95% CI, 16.06 to 25.44 pg/mL; p = 0.04). The difference was primarily due to the patients with active disease, who had significantly higher levels of 8-isoprostane (mean, 111.4 pg/mL; 95% CI, 62.56 to 160.30 pg/mL; p < 0.001) compared to patients with nonactive disease (mean, 22.94 pg/mL; 95% CI, 15.89 to 29.99 pg/mL) or healthy subjects. 8-Isoprostane levels in patients with nonactive disease did not differ from those in healthy subjects (p > 0.05). In the patients with sarcoidosis, 8-isoprostane levels were positively correlated with sACE level (p < 0.0001, r = 0.69), but was not correlated with serum calcium level or pulmonary function test values. CONCLUSIONS: Our data suggest that 8-isoprostane levels are increased in the expired breath condensate of patients with sarcoidosis and might serve as an index of disease activity.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=15006961 !0012-3692 (Print) Journal Article15006961UDepartment of Pneumonology, Army General Hospital of Athens, Greece. kpsazakis@hol.gr ~?}Bodini, A. Peroni, D. Vicentini, L. Loiacono, A. Baraldi, E. Ghiro, L. Corradi, M. Alinovi, R. Boner, A. L. Piacentini, G. L.2004aExhaled breath condensate eicosanoids and sputum eosinophils in asthmatic children: a pilot study26-31Pediatr Allergy Immunol151WAdolescent Altitude Asthma/*immunology/therapy Breath Tests/*methods Child Cysteine/analysis/immunology Eicosanoids/*analysis/immunology Eosinophils/*immunology Female Humans Leukotriene B4/analysis/immunology Leukotrienes/analysis/immunology Male Pilot Projects Prostaglandins A/analysis/immunology Pyroglyphidae/immunology Sputum/*immunologyFebCysteinyl leukotrienes (cys-LTs), LTB4 and 8-isoprostane are increased in the exhaled breath condensate (EBC) from asthmatic patients. The aim of this study was to investigate whether the measurement of cys-LTs, LTB4 and 8-isoprostane in EBC can reflect the level of airway inflammation assessed by induced sputum in asthmatic children sensitized to house dust mite (HDM) during natural avoidance of HDM allergens. Twelve children were evaluated at the time of admission (T0) and after 3 months of stay (T1) at the Istituto Pio XII (Misurina, Italian Dolomites 1756 m). Sputum eosinophil percentage and measurement of cys-LTs, LTB4 and 8-isoprostanes in the breath condensate at T0 and T1 were evaluated. Eosinophil percentage in induced sputum was 8.5 +/- 1.1% at T0 and 3.5 +/- 0.4% at T1 (p = 0.011). Neutrophil percentage in sputum was 1.1 +/- 0.5% at T0 and 1.5 +/- 1.0% at T1 (ns). Cys-LTs mean level was 14.24 +/- 4.53 pg/ml at T0 and 4.65 +/- 0.68 pg/ml at T1 (p = 0.0125). LTB4 level was 2.36 +/- 0.19 pg/ml at T0 and 2.41 +/- 0.23 pg/ml at T1 (ns). 8-Isoprostane level reduced from 17.47 +/- 3.18 pg/ml at T0 to 7.36 +/- 3.26 pg/ml at T1 (p = 0.003). This study show that exhaled cys-LTs and 8-isoprostane, as well as eosinophil percentage in induced sputum, are reduced after allergen avoidance in asthmatic children suggesting a potential application of EBC for the non-invasive evaluation of airway inflammation in asthma in allergic asthmatic children.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14998379 !0905-6157 (Print) Journal Article149983798Clinica Pediatrica, Universita di Verona, Verona, Italy.^~?[Rosias, P. P. Dompeling, E. Hendriks, H. J. Heijnens, J. W. Donckerwolcke, R. A. Jobsis, Q.2004:Exhaled breath condensate in children: pearls and pitfalls4-19Pediatr Allergy Immunol151YAdolescent Breath Tests/*methods Child Child, Preschool Cytokines/analysis Eicosanoids/analysis Extravascular Lung Water/*chemistry/*immunology Glutathione/analysis Humans Hydrogen Peroxide/analysis Immunoglobulin E/analysis Infant Lung Diseases/*immunology Malondialdehyde/analysis Oxidative Stress/physiology Reactive Nitrogen Species/analysisFebExhaled breath condensate (EBC) is a rapidly growing field of research in respiratory medicine. Airway inflammation is a central feature of chronic lung diseases, like asthma, cystic fibrosis, bronchopulmonary dysplasia and primary ciliary dyskinesia. EBC may be a useful technique for non-invasive assessment of markers of airway inflammation. The non-invasive character of EBC "inflammometry" and the general lack of appropriate techniques makes it particularly interesting for paediatrics. We provide a detailed update on the methods currently used for EBC collection and measurement of mediators. We emphasize on paediatric data. The apparent simplicity of the EBC method must not be overstated, as numerous methodological pitfalls have yet to overcome. Comparison and interpretation of data on this rapidly growing field of research is mainly hampered by the lack of standardization and the lack of specific high-sensitivity immunochemical or colorimetric assays. The initiative of the European Respiratory Society to institute a task force on this topic is a first step towards a uniform technique of EBC. Meanwhile, when using this technique or when interpreting research data, one should be fully aware of the possible methodological pitfalls.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14998377 (0905-6157 (Print) Journal Article Review14998377Division of Paediatric Respiratory Medicine, Department of Paediatrics, University Hospital Maastricht, Maastricht, The Netherlands. p.rosias@orbisconcern.nl~?WMontuschi, P. Ragazzoni, E. Valente, S. Corbo, G. Mondino, C. Ciappi, G. Ciabattoni, G.2003\Validation of 8-isoprostane and prostaglandin E(2) measurements in exhaled breath condensate502-7 Inflamm Res5212Breath Tests/*methods Chromatography, High Pressure Liquid Dinoprostone/*analysis/blood Female Humans Isoprostanes/*analysis/blood Lung Diseases/blood/*diagnosis/metabolism Male Middle Aged Radioimmunoassay Reproducibility of Results Research Support, Non-U.S. Gov't RespirationDecOBJECTIVE: To qualitatively validate radioimmunoassays for 8-isoprostane and prostaglandin (PG) E(2) in exhaled breath condensate. 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 (RP-HPLC). The eluted fractions were assayed for 8-isoprostane-like immunoreactivity and PGE(2)-like immunoreactivity by radioimmunoassays. In addition, simultaneous measurements of exhaled breath condensate unextracted samples with two anti-8-isoprostane and anti-PGE(2) sera with different cross-reactivity were performed. RESULTS: A single peak of 8-isoprostane-like immunoreactivity and PGE(2)-like immunoreactivity co-eluting with 8-isoprostane (retention time: 13 min) and PGE(2) (retention time: 21 min) standards, respectively, was identified by radioimmunoassays. Testing with two different antisera showed similar results for both 8-isoprostane-like immunoreactivity (limits of agreement = 4.5 pg/ml and - 4.1 pg/ml, n = 12) and PGE(2)-like immunoreactivity (limits of agreement = 6.1 pg/ ml and - 6.1 pg/ml, n = 12). CONCLUSION: This study provides evidence for the specificity of the radioimmunoassays for 8-isoprostane and PGE(2) in exhaled breath condensate. This is critical for proposing these markers as a non-invasive way for monitoring airway inflammation.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14991078 01023-3830 (Print) Clinical Trial Journal Article14991078Department of Pharmacology, School of Medicine, Catholic University of the Sacred Heart, L go F Vito, 00168, Rome, Italy. pmontuschi@rm.unicatt.itL~?`Van Hoydonck, P. G. Wuyts, W. A. Vanaudenaerde, B. M. Schouten, E. G. Dupont, L. J. Temme, E. H.2004YQuantitative analysis of 8-isoprostane and hydrogen peroxide in exhaled breath condensate189-92 Eur Respir J232Adult Belgium *Breath Tests F2-Isoprostanes/*analysis Humans Hydrogen Peroxide/*analysis Immunoenzyme Techniques Lung Diseases/diagnosis Male Middle Aged Predictive Value of Tests Reagent Kits, Diagnostic Research Support, Non-U.S. Gov't Smoking/*adverse effectsFebAExhaled breath condensate (EBC) provides a noninvasive means of sampling the lower respiratory tract. Collection of EBC might be useful in the assessment of airway oxidative stress in smokers. The aim of this study was to determine 8-isoprostane and hydrogen peroxide levels in EBC, and, in addition, to investigate the reproducibility of these measurements. EBC samples were collected from 12 healthy male smokers at three time points within 1 week. 8-isoprostane and H2O2 were measured in nonconcentrated EBC using immunochemical and colorimetric assays, respectively. 8-isoprostane and H2O2 were detected in only 36 and 47% of all EBC samples, respectively. It was not possible to calculate the within-subject variation in a reliable manner since only three of the 12 smokers exhibited detectable 8-isoprostane concentrations on all three occasions (mean 4.6 pg x mL(-1); range 3.9-7.7 pg x mL(-1)), whereas H2O2 could not be detected on all three occasions in any of the smokers. Spiking experiments revealed a recovery of 83.5-109.5% for 8-isoprostane and 69.9-129.0%, for H2O2 in fresh EBC samples. It was concluded that levels of 8-isoprostane and hydrogen peroxide cannot be reproducibly assessed in exhaled breath condensate from healthy smokers because of their low concentration and/or the lack of sensitivity of the available assays.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14979489 !0903-1936 (Print) Journal Article14979489lDept of Public Health, Division of Nutritional Epidemiology, Catholic University of Leuven, Leuven, Belgium.v~? Rahman, I.2004WReproducibility of oxidative stress biomarkers in breath condensate: are they reliable?183-4 Eur Respir J232:Adult Biological Markers/analysis *Breath Tests F2-Isoprostanes/*analysis Humans Hydrogen Peroxide/*analysis Immunoassay Lipid Peroxidation/*physiology Lung Diseases/*diagnosis/physiopathology Male Mass Fragmentography Oxidative Stress/*physiology Predictive Value of Tests Smoking/*adverse effects/physiopathologyFebfhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14979486 #0903-1936 (Print) Comment Editorial14979486 ,~?)Vass, G. Huszar, E. Barat, E. Horvath, I.2003I[Exhaled breath condensate and its analysis--a new method in pulmonology]2517-24 Orv Hetil14451Asthma/diagnosis/metabolism Biological Markers/analysis Breath Tests/*methods Bronchiectasis/diagnosis/metabolism Cystic Fibrosis/diagnosis/metabolism *Dinoprost/*analogs & derivatives English Abstract F2-Isoprostanes/analysis Humans Hydrogen Peroxide/analysis Oxidative Stress Pulmonary Disease, Chronic Obstructive/diagnosis/metabolism Respiratory Distress Syndrome, Adult/diagnosis/metabolism Respiratory Tract Diseases/*diagnosis/*metabolism Smoking/metabolismDec 219In the middle of the nineties a new, non-invasive method for investigation of the lung aroused the interest of many researchers: the exhaled breath condensate. It shows the extent of the interest that in the last five years more than 80 original articles have been published in this theme. Many substances are found in the expired breath which are detectable in the liquid that we obtain by cooling (= condensing) the exhaled breath. The advantages of this method are that it is non-invasive, convenient, it could be performed with mechanically ventilated patients as well as with children. The most studied substance is the hydrogen-peroxide, which is the marker of oxidative stress, and its level in condensate is elevated in numerous inflammatory diseases. 8-isoprostane was also studied a lot, which is another marker of oxidative stress. Numerous substances could be even measured in condensate, so the decay-product of nitric-oxide (nitrite, nitrate, nitrotyrosine), further nitrosothiol, adenosine, ammonia, different ions, leukotrienes, cytokines; recently even other feature of condensate is examined, such as its pH. The different mediators could help us to know better the diseases, support the diagnosis, follow the treatment or the disease. In this study the authors attempt to present the most important knowledge till now.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14974158 (0030-6002 (Print) Journal Article Review14974158cA kilegzett levego kondenzalasa es a kondenzatum elemzo vizsgalata. Uj modszer a tudogyogyaszatban.aOrszagos Koranyi Tbc es Pulmonologiai Intezet, Korelettani Osztaly, Budapest. vassgeza@koranyi.hu~?gZacharasiewicz, A. Wilson, N. Lex, C. Li, A. Kemp, M. Donovan, J. Hooper, J. Kharitonov, S. A. Bush, A.2004BRepeatability of sodium and chloride in exhaled breath condensates273-5Pediatr Pulmonol373Adult Asthma/*diagnosis/pathology Breath Tests Child Chlorides/*analysis Cystic Fibrosis/*diagnosis/pathology Humans Pneumonia/*diagnosis/pathology Reproducibility of Results Sodium/*analysisMar?Exhaled breath condensate (EBC) has been proposed as a noninvasive tool to study airway inflammation. The reproducibility of breath condensates was recently questioned. We therefore measured sodium and chloride concentrations in EBC and assessed the repeatability of these measurements in healthy adults and children with airway disease. We investigated technical repeatability and within-day repeatability in five healthy adults, and compared these results with those of 10 asthmatic children and 9 children with cystic fibrosis (CF). We also assessed within-period repeatability in the healthy controls. We report that the variability of measurements was similar for within sample, within day, and between visits, for both normals and children with asthma and CF, and that the major source of variability of sodium and chloride measurements is restricted by the reproducibility of the measurement assay method used. The wide use of EBC is more likely to depend on the development of highly sensitive and reproducible assays, rather than further refinements of the collection technique.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14966822 08755-6863 (Print) Clinical Trial Journal Article14966822KDepartment of Respiratory Paediatrics, Royal Brompton Hospital, London, UK. ~?jZanconato, S. Carraro, S. Corradi, M. Alinovi, R. Pasquale, M. F. Piacentini, G. Zacchello, F. Baraldi, E.2004gLeukotrienes and 8-isoprostane in exhaled breath condensate of children with stable and unstable asthma257-63J Allergy Clin Immunol1132Adolescent Androstadienes/therapeutic use Anti-Asthmatic Agents/therapeutic use Asthma/drug therapy/metabolism/*physiopathology Biological Markers Breath Tests Budesonide/therapeutic use Child Comparative Study Cysteine/*metabolism *Dinoprost/*analogs & derivatives F2-Isoprostanes/*metabolism Female Forced Expiratory Volume Humans Leukotrienes/*metabolism Male Nitric Oxide/metabolism *Oxidative Stress Research Support, Non-U.S. Gov't *Severity of Illness IndexFebBACKGROUND: Cysteinyl-leukotrienes (cys-LTs) and 8-isoprostane are biomarkers of airway inflammation and oxidative stress. OBJECTIVE: The aim of this study was to evaluate cys-LT and 8-isoprostane levels in exhaled breath condensate (EBC) of children with different degrees of asthma severity. METHODS: EBC was collected from 14 steroid-naive children with mild persistent asthma, 13 children with stable mild- to-moderate persistent asthma treated with inhaled corticosteroids (ICS), 9 ICS-treated children with unstable asthma, and 19 healthy children. RESULTS: In the three groups of asthmatic children, EBC concentrations of cys-LTs and 8-isoprostane were significantly higher than in control children (steroid-naive asthmatic children: cys-LTs median, 10.8 pg/mL, P <.001, 8-isoprostane, 16.2 pg/mL, P <.001; ICS-treated stable asthmatic children: cys-LTs, 12.7 pg/mL, P <.001, 8-isoprostane, 18.1 pg/mL, P <.001; children with unstable asthma: cys-LTs, 106.0 pg/mL, P <.01, 8-isoprostane, 29.7 pg/mL, P <.01; control children: cys-LTs, 4.3 pg/mL, 8-isoprostane, 3.5 pg/mL). Cys-LT levels were higher in children with unstable asthma than in the other two asthmatic groups (P <.05). FE(NO) levels were significantly higher in steroid-naive and in children with unstable asthma compared with ICS-treated children with stable asthma (P <.01). CONCLUSIONS: Our study shows that EBC cys-LTs and 8-isoprostane concentrations are higher in asthmatic children than in healthy control children, with scattered values in patients with unstable asthma. These findings suggest that EBC eicosanoid measurement may have useful clinical implications for investigating phenotype differences among asthmatic patients.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14767439 00091-6749 (Print) Clinical Trial Journal Article14767439>Department of Pediatrics, University of Padova, Padova, Italy. ~?Rahman, I. Kelly, F.2003`Biomarkers in breath condensate: a promising new non-invasive technique in free radical research1253-66Free Radic Res3712Biological Markers Breath Tests/*methods Free Radicals/*analysis Humans Lung Diseases/*diagnosis *Oxidative Stress Research Support, Non-U.S. Gov'tDec@Oxidative stress is associated with a range of inflammatory lung diseases including asthma, adult respiratory distress syndrome, idiopathic pulmonary fibrosis, pneumonia, lung transplantation, chronic obstructive pulmonary disease, cystic fibrosis, bronchiectasis and lung cancer. Increased concentrations of reactive oxygen species (ROS) in the airways of such patients are reflected by elevated concentrations of oxidative stress markers in the breath, airways, lung tissue and blood. Traditionally, the measurement of these biomarkers has involved invasive procedures to procure the samples, or examine the compartments. As a consequence, there is a need for less invasive approaches to measure oxidative stress. Analysis of breath hydrocarbons has partly fulfilled this need, however only gas phase volatile constituents can be assessed by this approach. The collection of exhaled breath condensate (EBC) is a simple, non-invasive approach, which comprehensively samples the lower respiratory tract. It is currently used as a research and diagnostic tool in the free radical field, yielding information on redox disturbance and the degree and type of inflammation in the lung. With further technical developments, such an approach may ultimately have a role in the clinic, in helping to diagnose specific lung diseases. EBC can be exploited to assess a spectrum of potential biomarkers, thus generating a "finger print" characteristic of the disease. By assessing the nature of oxidative stress in this manner, the most appropriate therapy can be selected and the response to treatment monitored.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14753750 (1071-5762 (Print) Journal Article Review14753750Respiratory Medicine, ELEGI, Colt Research Laboratories, MRC Centre for Inflammation Research, The University of Edinburgh Medical School, Wilkie Building, Teviot Place, Edinburgh EH8 9AG, UK. irfan.rahman@ed.ac.uk~?HGessner, C. Kuhn, H. Toepfer, K. Hammerschmidt, S. Schauer, J. Wirtz, H.2004cDetection of p53 gene mutations in exhaled breath condensate of non-small cell lung cancer patients215-22 Lung Cancer432TActins/genetics Aged Aged, 80 and over Breath Tests Carcinoma, Non-Small-Cell Lung/*genetics Comparative Study DNA Damage *DNA Mutational Analysis DNA, Neoplasm/genetics Female Genes, p53/*genetics Humans Lung Neoplasms/*genetics Male Middle Aged *Point Mutation Polymerase Chain Reaction Sensitivity and Specificity Smoking/adverse effectsFeb-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 beta-actin gene fragment was used to detect human DNA in each of the EBC samples. In 65.7% of the samples, the beta-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.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14739043 00169-5002 (Print) Clinical Trial Journal Article14739043Department of Respiratory and Critical Care Medicine, University of Leipzig, Johannisallee 32, 04103 Leipzig, Germany. gesc@medizin.uni-leipzig.de ~?KGarey, K. W. Neuhauser, M. M. Robbins, R. A. Danziger, L. H. Rubinstein, I.2004MMarkers of inflammation in exhaled breath condensate of young healthy smokers22-6Chest1251.Adult *Breath Tests Chemotaxis, Leukocyte Female Humans Inflammation Mediators/*analysis Interleukin-1/analysis Male Neutrophils/physiology Nitrites/analysis Proteins/analysis Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Smoking/*metabolism Tumor Necrosis Factor-alpha/analysisJanINTRODUCTION: Although a strong correlation exists between long-term cigarette smoking, pulmonary inflammation, and COPD, efforts to identify populations at risk of acquiring COPD have so far been unsuccessful. To this end, noninvasive detection and monitoring of biomarkers of pulmonary inflammation in young healthy smokers may assist in this task. STUDY OBJECTIVES: The purpose of this study was to determine the concentrations of total protein, nitrites, interleukin (IL)-1beta, and tumor necrosis factor (TNF)-alpha, and neutrophil chemotactic activity in exhaled breath condensate (EBC) collected from healthy college student smokers and nonsmokers. DESIGN: EBC was collected from 20 volunteers (9 nonsmokers and 11 smokers) during tidal breathing for 20 min. EBC was also collected from smokers 30 min after smoking one filtered cigarette. The concentrations of total protein, nitrite, IL-1beta, and TNF-alpha in EBC was determined by enzyme-linked immunosorbent assay. Neutrophil chemotactic activity in EBC was determined in vitro using the blind-well technique. RESULTS: The concentrations of total protein and nitrite, and neutrophil chemotactic activity were significantly higher in EBC of smokers in comparison to nonsmokers (p < 0.05). The concentrations of total protein and nitrite in the condensate of smokers did not change significantly after smoking one cigarette. The concentrations of IL-1beta and TNF-alpha in EBC were similar in nonsmokers and smokers. CONCLUSIONS: Concentrations of certain inflammatory mediators and neutrophil chemotactic activity are increased in EBC of young healthy smokers. Collection and analysis of EBC may assist in early detection of cigarette smoke-induced pulmonary inflammation and identifying populations at risk for acquiring COPD.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14718416 !0012-3692 (Print) Journal Article14718416<University of Houston College of Pharmacy, Houston, TX, USA.~?]Siniachenko, O. V. Kazakov, V. N. Faerman, A. A. Pesin Ia, M. Belokon, T. M. Fainerman, V. B.2003Z[Physical and chemical properties of the condensate of breathed-out air in healthy people] 23-4, 33-5Klin Lab Diagn11Adult Age Factors Breath Tests/methods English Abstract Female Humans Hydrogen Peroxide/*analysis Male Nitric Oxide/*analysis Pulmonary Surfactants/*chemistry Rheology Sex Factors SpectrophotometryNovThe physical-and-chemical condition of condensate of breathed-out moisture (CBM) was studied by the computer-aided analysis of axis-symmetric drops' form in healthy subjects; such condition was found to depend on sex, age and smoking-addiction of the examined persons. The surface tension of the mentioned moisture correlated with its viscoelastic index and with relaxation. The metabolites of nitric oxide, lipids, urea, lactic acid (not uric acid) and of hydrogen peroxide were found to influence the physical-and-chemical parameters of respiratory moisture. Hydrogen peroxide had a reverse correlation with the CBM viscoelastic module. A status of the pulmonary surfactant can be evaluated by using the studied physical-and-chemical CBM parameters.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14708368 !0869-2084 (Print) Journal Article14708368QFiziko-khimicheskie svoistva kondensata vydykhaemogo vozdukha u zdorovykh liudei.~?9Bucchioni, E. Kharitonov, S. A. Allegra, L. Barnes, P. J.2003SHigh levels of interleukin-6 in the exhaled breath condensate of patients with COPD1299-302 Respir Med9712Adult Aged Biological Markers/analysis Breath Tests/methods Female Humans Immunoenzyme Techniques/methods/standards Interleukin-6/*analysis Male Middle Aged Pulmonary Disease, Chronic Obstructive/*metabolism Reproducibility of Results Smoking/metabolismDeceBACKGROUND: 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 exsmokers 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 +/- 0.1 pg/ml; P < 0.0001) than in the healthy non-smokers (4.9 +/- 0.2 pg/ml) with a correlation in this group between age and IL-6 levels (r = 0.597; P < 0.05). CONCLUSIONS: The increased IL-6 levels in exhaled breath condensate may reflect airway inflammation in patients with COPD.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14682411 J0954-6111 (Print) Clinical Trial Controlled Clinical Trial Journal Article14682411Institute of Respiratory Diseases, University of Milan, Ospedale Maggiore IRCCS, Via F. Sforza, 35, Milano 20125, Italy. e.bucchioni@ic.ac.ukT~?xVaughan, J. Ngamtrakulpanit, L. Pajewski, T. N. Turner, R. Nguyen, T. A. Smith, A. Urban, P. Hom, S. Gaston, B. Hunt, J.2003QExhaled breath condensate pH is a robust and reproducible assay of airway acidity889-94 Eur Respir J226Acid-Base Equilibrium/physiology Adolescent Adult Biological Markers Breath Tests/*methods Female Humans Hydrogen-Ion Concentration Lung Diseases/*diagnosis/physiopathology Male Middle Aged Respiratory System/*physiopathologyDecaExhaled breath condensate (EBC) pH is low in several lung diseases and it normalises with therapy. The current study examined factors relevant to EBC pH monitoring. Intraday and intraweek variability were studied in 76 subjects. The pH of EBC collected orally and from isolated lower airways was compared in an additional 32 subjects. Effects of ventilatory pattern (hyperventilation/hypoventilation), airway obstruction after methacholine, temperature (-44 to +13 degrees C) and duration of collection (2-7 min), and duration of sample storage (up to 2 yrs) were examined. All samples were collected with a disposable condensing device, and de-aerated with argon until pH measurement stabilised. Mean EBC pH (n=76 subjects, total samples=741) was 7.7+/-0.49 (mean+/-SD). Mean intraweek and intraday coefficients of variation were 4.5% and 3.5%. Control of EBC pH appears to be at the level of the lower airway. Temperature of collection, duration of collection and storage, acute airway obstruction, subject age, saliva pH, and profound hyperventilation and hypoventilation had no effect on EBC pH. The current authors conclude that in health, exhaled breath condensate pH is slightly alkaline, held in a narrow range, and is controlled by lower airway source fluid. Measurement of exhaled breath condensate pH is a simple, robust, reproducible and relevant marker of disease.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14680074 !0903-1936 (Print) Journal Article14680074iDivision of Pediatric Respiratory Medicine, University of Virginia, Charlottesville, Virginia 22908, USA. ~?RRysz, J. Kasielski, M. Apanasiewicz, J. Krol, M. Woznicki, A. Luciak, M. Nowak, D.2004aIncreased hydrogen peroxide in the exhaled breath of uraemic patients unaffected by haemodialysis158-63Nephrol Dial Transplant191Adult Breath Tests Female Humans Hydrogen Peroxide/*analysis Kidney Failure, Chronic/*metabolism/physiopathology/therapy Male Middle Aged Renal Dialysis Research Support, Non-U.S. Gov't Uremia/*metabolism/physiopathology/therapyJanBACKGROUND: Uraemia is accompanied by conditions favouring the rise of H2O2 activity in body fluids. This results from the increased release of H2O2 by polymorphonuclear leukocytes and decreased plasma glutathione peroxidase activity. The purpose of this study was to determine if patients on chronic haemodialysis (HD) exhale more H2O2 than healthy individuals, and if dialysis affects breath H2O2 content. METHODS: We studied 29 chronic HD patients (mean age 49 +/- 11 years) and 40 healthy persons (mean age 44 +/- 9 years). H2O2, which is volatile, was measured fluorimetrically with the homovanillic acid method in the exhaled breath condensate (EBC) of the study cohort. EBC was collected immediately before and after the HD session and also at 20 and 60 min of HD treatment (n = 14) and once in controls. Peak expiratory flow (PEF), white blood cell (WBC) count, PaO(2) and circulatory cyclic guanosine monophosphate (cGMP), Il-6 and Il-8 concentrations were measured concomitantly. Finally, H2O2 diffusion through the dialyser cuprophane membrane was determined in an in vitro experiment. RESULTS: At baseline, EBC H2O2 concentration was 22 times higher in HD patients than in controls (2.92 +/- 4.64 vs 0.16 +/- 0.13 microM, P < 0.001). Although the maximum decrease in PEF (431 +/- 52 vs 398 +/- 56 l/min, P < 0.01) and WBC count (6.72 +/- 1.02 vs 3.82 +/- 1.51 x 10(3)/ microl, P < 0.01) occurred at 20 min after the start of HD, no significant changes in breath H2O2 levels were noted throughout the session. Plasma IL-6 and IL-8 levels remained unchanged whereas cGMP rose 1.3 times at 60 min (P < 0.01). In vitro, H2O2 rapidly diffused through the cuprophane membrane. CONCLUSION: Chronic HD patients exhale more H2O2 than healthy subjects. Although no change of breath H2O2 concentration was observed during HD, as H2O2 easily diffuses through the dialyser membrane, it is not possible to rule out that HD stimulates H2O2 generation.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14671051 !0931-0509 (Print) Journal Article14671051xDepartment of Internal Medicine and Dialysotherapy Medical University of Lodz, Mazowiecka Str. 6/8, 92-215 Lodz, Poland.~?BGel'tser, B. I. Peteshova, E. E. Kochetkova, E. A. Eliseeva, E. V.2003[The test for nitric oxide metabolites in exhaled air condensate as a method of assessing NO-reactivity of the airways in patients with bronchial asthma]91-4Ter Arkh7510Asthma/diagnosis/metabolism/*physiopathology Breath Tests Bronchodilator Agents/diagnostic use English Abstract Fenoterol/diagnostic use Humans Nitrates/*analysis Nitric Oxide/*metabolism Nitrites/*analysis *Respiration Respiratory Function Tests]AIM: To estimate airways NO-reactivity in response to their stimulation with fenoterol in different clinical forms of bronchial asthma (BA). MATERIAL AND METHODS: The study included 73 patients with BA: mild disease was in 21, moderate--in 24 and severe--in 28 patients. Severe BA patients were divided into two subgroups: with a stable course (n = 15) and unstable course (n = 13). NO-producing function of the airways was estimated by concentration of stable NO-metabolites (mNO)(NO2, NO3) in exhaled air condensate. RESULTS: Spontaneous NO-producing activity of the airways increases and reaches maximum in severe unstable asthma. Fenoterol-stimulated NO-production was minimal in mild BA while the most significant augmentation of mNO was observed in unstable BA. Basal level of mNO and velocity parameters of external respiration function correlated. CONCLUSION: Estimation of NO-reactivity of the airways in the test with fenoterol with calculation of the index of airways NO-reactivity provides additional information about respiratory system condition in BA patients which may be used in clinical pulmonology.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14669617 !0040-3660 (Print) Journal Article14669617Opredelenie metabolitov oksida azota v kondensate vydykhaemogo vozdukh kak sposob otsenki NO-reaktivnosti dykhatel'nykh putei u bol'nykh bronkhial'noi astmoi. ~?cvan Beurden, W. J. van den Bosch, M. J. Janssen, W. C. Smeenk, F. W. Dekhuijzen, P. N. Harff, G. A.2003EFluorimetric analysis of hydrogen peroxide with automated measurement637-43Clin Lab4911-12 Adult Aged Aged, 80 and over Area Under Curve Breath Tests/instrumentation/*methods Female Fluorometry/*methods Humans Hydrogen Peroxide/*analysis Male Middle Aged Pulmonary Disease, Chronic Obstructive/*diagnosis Reproducibility of Results Sensitivity and SpecificityIn the pathophysiology of chronic obstructive pulmonary disease (COPD) oxidative stress plays an important role, which can be determined by measuring hydrogen peroxide. Hydrogen peroxide can be measured fluorimetrically in exhaled breath condensate (EBC), however, not standardized. The objective of this study was to investigate the sensitivity and reproducibility of measuring the hydrogen peroxide concentration in EBC of COPD patients using an automated flow injection device with varying flow rates and measurements. METHODS: 10 microl p-hydroxyphenylacetic acid (1.0 mmol/l) and 10 microl horseradish peroxidase (15 mU/l) were manually added to several hydrogen peroxide containing solutions and EBC of patients suffering from COPD. The fluorescence of the reaction product was measured with an automated sampler, flow injection and scanning fluorescence detector, excitation wave 295 nm, emission wave 405 nm, at different flow rates. The degree of fluorescence was expressed as either the area under the curve or the peak value. RESULTS: A flow rate of 1 ml/min gave the best results. There were no significant differences in calibration curves or detection limits using area under the curve or peak value (respectively 0.007 and 0.005 micromol/l) (flow rate 1.0 ml/min). The mean volume of EBC was 2.8 ml, the mean hydrogen peroxide concentration in the patient group was 0.2 micromol/l and the standard deviation of duplication 0.009 micromol/. CONCLUSIONS: The low detection limit may be explained by using flow injection, because it measures the fluorescence over a period of time. It is important to choose an appropriate flow rate. There is no difference in the detection limit between measuring the fluorescence as area under the curve or as peak value.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14651334 !1433-6510 (Print) Journal Article14651334lDepartment of Pulmonary Diseases, Catharina Hospital Eindhoven, The Netherlands. wendy.beurdenvan@wanadoo.nls~?.Latzin, P. Beck, J. Bartenstein, A. Griese, M.2003FComparison of exhaled breath condensate from nasal and oral collection505-10 Eur J Med Res811Adult Biological Markers Breath Tests Child Comparative Study Cystic Fibrosis/immunology/*metabolism Exhalation Humans Hydrogen Peroxide/*metabolism *Mouth Nitrates/metabolism Nitrites/metabolism *NoseNov 12 BACKGROUND: Analysis of exhaled breath condensate may provide new insights into pulmonary inflammatory processes. A new collection method via suction of nasally expired air especially suitable for younger children was presented recently. Here we compare this nasal suction method with the more widely used oral collection method regarding the amount of condensate collected as well as the concentrations of hydrogen peroxide (H2O2), nitrite and nitrate, respectively. MATERIALS AND METHODS: Exhaled breath condensate was collected from 11 healthy adults for the measurements of the amount of condensate and H2O2 concentration and from 17 children for the measurements of nitrite and nitrate. Condensate was collected via nasal suction and oral exhalation from each subject. RESULTS: Overall, no differences between both collection methods were found for all variables assessed except the concentration of H2O2, whereas the latter closely correlated (Spearman r = 0.88, p = 0.0007) between both collection methods. No correlation was found for the amount of condensate collected and the concentration of nitrite and nitrate. The Bland-Altman limits of agreement scattered over a wide range with clinical impact, proving significant differences between both collection methods for all variables measured. CONCLUSIONS: Although nasal and oral collection method proved again suitable for the collection of exhaled breath condensate, the variability of the results obtained precludes the interchangeable usage of the inflammatory markers assessed here.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14644706 00949-2321 (Print) Clinical Trial Journal Article146447068CF-Ambulanz, Lindwurmstrasse 4, D-80337 Munich, Germany.~?cGessner, C. Hammerschmidt, S. Kuhn, H. Seyfarth, H. J. Sack, U. Engelmann, L. Schauer, J. Wirtz, H.2003<Exhaled breath condensate acidification in acute lung injury1188-94 Respir Med9711[Acute Disease Ammonia/analysis Amylases/metabolism Biological Markers/analysis Breath Tests/methods Carbon Dioxide/analysis Carbonic Acid/analysis Enzyme-Linked Immunosorbent Assay/methods Female Humans Hydrogen-Ion Concentration Interleukins/analysis Lactates/analysis Male Middle Aged Pneumonia/*diagnosis Respiration, Artificial/adverse effectsNovLung 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, pCO2, HCO3-, 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 < 0.0001). NH4+, lactate, HCO3-, pCO2, 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 < 0.0001, EBC IL-8: r = -0.68, P < 0.0001) but not of systemic inflammation (serum IL-6, serum IL-8) and with indices of severity of lung injury (Murray's Lung Injury Severity Score; r = -0.73, P < 0.0001, paO2/FiO2; r = 0.54, P < 0.001). Among factors potentially contributing to pH of EBC, EBC-lactate and EBC-NH4+ 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.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14635972 !0954-6111 (Print) Journal Article14635972Department of Internal Medicine, Pulmonary Medicine, Critical Care and Cardiology, University of Leipzig, Johannisallee 32, Leipzig 04103, Germany. gesc@medizin.uni-leipzig.de ;~?EKostikas, K. Papatheodorou, G. Psathakis, K. Panagou, P. Loukides, S.2003IProstaglandin E2 in the expired breath condensate of patients with asthma743-7 Eur Respir J225Adult Asthma/*metabolism/physiopathology *Breath Tests Bronchial Provocation Tests Dinoprostone/*metabolism Forced Expiratory Volume Histamine/diagnostic use Humans Reproducibility of Results Respiratory System/metabolism Smoking/metabolism/physiopathologyNovxInhaled prostaglandin (PG)E2 has been found to cause bronchodilation in asthmatics, although it does not have bronchodilative effects in normal subjects. The aim of this study was to investigate the levels of PGE2 in the expired breath condensate of patients with asthma, the possible contribution of smoking habit to its levels and the possible relationship between PGE2 and the degree of bronchial hyperresponsiveness, as assessed by the provocation dose of histamine causing a 20% fall in forced expiratory volume in one second (FEV1) (PD20). A total of 30 mild asthmatics (15 smokers, all steroid-naive, FEV1 88+/-6 (%+/-SD)) and 20 healthy control subjects (10 smokers) were studied. Histamine challenge testing was performed in all subjects and the PD20 was determined. The results showed that asthmatic smokers had significantly higher values of PGE2 compared to asthmatic nonsmokers and control subjects (40+/-21 versus 14.5+/-4.5 versus 11.7+/-3 pg x mL(-1), respectively). Further analysis showed that PGE2 levels were significantly higher in asthmatic smokers compared to smoker and nonsmoker controls (40+/-21 versus 11.6+/-2 versus 11.7+/-4 pg x mL(-1), respectively). No significant difference was observed between asthmatic nonsmokers and both control smokers and control nonsmokers. No significant correlation was found between PGE2 levels and PD20 in all groups of asthmatics, irrespective of smoking habit. In conclusion, the elevation of prostaglandin E2 in the expired breath condensate of patients with asthma is mainly attributed to smoking habit and prostaglandin E2 levels do not predict the degree of bronchial hyperresponsiveness.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14621079 !0903-1936 (Print) Journal Article14621079VPneumonology and Clinical Research Dept, Athens Army General Hospital, Athens, Greece. ~?Hitka, P. Vizek, M. Wilhelm, J.2003:Hypoxia and reoxygenation increase H2O2 production in rats585-92 Exp Lung Res298Animals Anoxia/*metabolism/pathology Breath Tests/methods Hydrogen Peroxide/*metabolism Lung/*drug effects/metabolism Male Oxygen/*pharmacology Rats Rats, Wistar Reactive Oxygen Species Research Support, Non-U.S. Gov't RespirationDecTo test the effect of transition from sustained hypoxia to normoxia on production of reactive oxygen species (ROS) in lungs, the authors measured hydrogen peroxide (H(2)O(2)) output in the expired air of rats breathing hypoxic, normoxic, and hyperoxic gas mixtures at the end of exposure to 72 hours of hypoxia. Twenty-one male Wistar rats (200 to 280 g) were randomly assigned to 1 of 3 groups. First two groups (experimental) were kept for 3 days in normobaric hypoxic chamber (F(1)O(2) 0.1), rats of the third group (controls) breathed air. The rats were then anesthetized, intubated, placed in the plethysmograph, and their ventilation measured. Two periods of exhaled breath condensate (EBC) collection, each lasting 1 hour, were then performed to assay H(2)O(2) output. The controls breathed during both samplings air, the first experimental group breathed during first sampling period hypoxic mixture (F(1)O(2) 0.1; SH-H measurement) and then, during second period, air (SH-H-A measurement), the second experimental group breathed first air (SH-A measurement) and then hyperoxic mixture (F(1)O(2) 1.0; SH-A-O(2) measurement). Concentration of H(2)O(2) in the EBC was assayed by chemiluminescence. H(2)O(2) production in the control group was low and similar in both measurements (20+/-10 and 13+/-5 pmol/h, mean+/-SEM). Exposure to 72 hours of hypoxia increased the H(2)O(2) production to 105+/-18 pmol/h (SH-H). Transition from hypoxia to normoxia resulted in an increase in the H(2)O(2) production (SH-A 421+/-24 pmol/h, and SH-H-A 366+/-19 pmol/h). Following transition from air breathing to hyperoxia did not affect the H(2)O(2) production (SH-A-O(2) 373+/-25 pmol/h). The results showed that sustained hypoxia and transition from sustained hypoxia to normoxia increased H(2)O(2) formation in the lungs.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14594657 !0190-2148 (Print) Journal Article14594657Department of Pathophysiology, Charles University, Second Medical Faculty, and Centre for Experimental Cardiovascular Research, Prague, Czech Republic. patrikhitka@hotmail.com~?-Cheah, F. C. Darlow, B. A. Winterbourn, C. C.2003Problems associated with collecting breath condensate for the measurement of exhaled hydrogen peroxide from neonates on respiratory support338-41 Biol Neonate844*Breath Tests/instrumentation Continuous Positive Airway Pressure Humans Hydrogen Peroxide/*analysis Infant, Newborn Lung Diseases/diagnosis *Respiration, Artificial Specimen Handling/*methodsWe developed a bedside method for collecting exhaled breath condensate (EBC) from neonates who were ventilated or receiving nasal continuous positive airway pressure (CPAP) and analyzed their EBC for hydrogen peroxide levels. A sufficient volume for analysis could be collected over 25-40 min from neonates on the ventilator and nasal CPAP (medians 5.3 and 2.7 ml, respectively). There was no significant difference between hydrogen peroxide levels from neonates on a ventilator or CPAP (median 0.28 vs. 0.38 microM, p = 0.06) and these were no different from a background with the ventilator or CPAP system alone (median for each 0.31 microM). The dilution of breath condensate by humidified gases plus the existence of background hydrogen peroxide resulted in this collecting setup being insufficiently sensitive to use for the detection of exhaled hydrogen peroxide in infants who were ventilated or on nasal CPAP.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14593246 !0006-3126 (Print) Journal Article14593246Free Radical Research Group, Department of Pathology, Christchurch School of Medicine and Health Sciences, University of Otago, Christchurch, New Zealand. fookchoe.cheah@chmeds.ac.nzu~? Effros, R. M.2003%Saving the breath condensate approach1129-32; author reply 1130-1Am J Respir Crit Care Med1689DAsthma/metabolism Breath Tests/*methods Glutathione/*analysis HumansNov 1fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14581290 1073-449X (Print) Comment Letter14581290S~? Dwyer, T. M.2003WCigarette smoke-induced airway inflammation as sampled by the expired breath condensate174-8 Am J Med Sci3264#*Breath Tests Emphysema Genetic Predisposition to Disease Humans Inflammation/diagnosis/*etiology/pathology Reactive Oxygen Species/analysis Research Support, Non-U.S. Gov't Respiratory Tract Diseases/diagnosis/*etiology/pathology Smoke/*adverse effects/analysis *Tobacco alpha 1-AntitrypsinOctQAirways exposed to smoke respond with inflammatory processes. The airway inflammation generally present in smokers causes persistent cough and phlegm production, reactive airway disease, and tissue infiltration by inflammatory cells. Although the short-term response may be protective, long-term pathological consequences include swelling of the airway epithelium, mucus hypersecretion, and increased airway reactivity characteristic of chronic bronchitis and chronic obstructive lung disease and the tissue destruction characteristic of emphysema. The natural history of these diseases is poorly understood, because human airway tissue is available for study only at autopsy, from surgical specimens, or from procedures such as bronchoscopy or thoracotomy. A noninvasive method of monitoring the inflammation is by analyzing expired breath condensate, which contains a diluted sample of airway surface liquid. The study of expired breath condensate may offer a more practical approach to sampling airway chemistry and make it possible to study the detailed inflammatory response to airborne particulates.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14557729 (0002-9629 (Print) Journal Article Review14557729Department of Physiology and Biophysics, University of Mississippi Medical Center, Jackson, Mississippi 39216-4505, USA. tdwyer@physiology.umsmed.edu ~?cCarpagnano, G. E. Kharitonov, S. A. Resta, O. Foschino-Barbaro, M. P. Gramiccioni, E. Barnes, P. J.20038-Isoprostane, a marker of oxidative stress, is increased in exhaled breath condensate of patients with obstructive sleep apnea after night and is reduced by continuous positive airway pressure therapy1386-92Chest1244Breath Tests *Continuous Positive Airway Pressure *Dinoprost/*analogs & derivatives Exhalation F2-Isoprostanes/*analysis Female Humans Male Middle Aged *Oxidative Stress Sleep Apnea, Obstructive/*metabolism/*therapyOctSTUDY OBJECTIVES: Obstructive sleep apnea (OSA) is characterized by recurrent apnea during sleep that may compromise oxidative balance. Oxidative stress is increased in the blood and in the airways of OSA patients. DESIGN: The aim of this study was to investigate whether oxidative stress is determined by nocturnal apneas and could be reduced by CPAP therapy, and whether there is a relation between local and systemic oxidative stress in these patients. PATIENTS AND METHODS: Eighteen patients with OSA (13 men; mean [+/- SD] age, 48 +/- 3 years) and 12 healthy age-matched and weight-matched subjects (8 men; mean age, 46 +/- 7 years) were recruited. 8-Isoprostane was measured in exhaled breath condensate and blood by a specific enzyme immunoassay. Measurements and results: Higher concentrations of 8-isoprostane were found in the morning exhaled condensate (9.5 +/- 1.9 pg/mL) and plasma (9.7 +/- 1.5 pg/mL) of OSA patients compared to healthy obese subjects (6.7 +/- 0.2 and 7.1 +/- 0.3 pg/mL, respectively; p < 0.0001). Elevated mean concentrations of exhaled 8-isoprostane were observed in the OSA patients at 8:00 AM (9.5 +/- 1.9 pg/mL) but not at 8:00 PM (7.6 +/- 0.8 pg/mL; p < 0.0005), and a significant reduction was seen after continuous positive airway pressure (CPAP) therapy (7.7 +/- 0.9 pg/mL; before treatment, 9.6 +/- 1.7 pg/mL; p < 0.005). A positive correlation was found between morning exhaled 8-isoprostane levels and the apnea-hypopnea index (r = 0.8; p < 0.0001), and 8-isoprostane levels and neck circumference (r = 0.6; p < 0.0001). CONCLUSIONS: These findings suggest that systemic and local oxidative stress are increased in OSA patients, and that they are higher after nocturnal apnea and reduced by CPAP therapy.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14555570 !0012-3692 (Print) Journal Article14555570CInstitute of Respiratory Diseases, University of Bari, Bari, Italy. ~?EKostikas, K. Papatheodorou, G. Psathakis, K. Panagou, P. Loukides, S.2003COxidative stress in expired breath condensate of patients with COPD1373-80Chest1244"Adult Aged Androstadienes/therapeutic use Anti-Inflammatory Agents/therapeutic use Breath Tests Bronchodilator Agents/therapeutic use Exhalation Humans Male Middle Aged *Oxidative Stress Pulmonary Disease, Chronic Obstructive/drug therapy/immunology/*metabolism Theophylline/therapeutic useOct7 OBJECTIVE: To evaluate the levels of hydrogen peroxide (H(2)O(2)) and 8-isoprostane in the expired breath condensate (EBC) of patients with COPD, and to assess the relationship between the above markers of oxidative stress and parameters expressing inflammatory process and disease severity. SETTING: Inpatient respiratory unit and outpatient clinic in tertiary care hospital. DESIGN: Cross-sectional study. PATIENTS: Thirty stable COPD patients (all smokers) with disease severity ranging from mild to severe. Ten subjects who were smokers with stage 0 disease (ie, at risk for COPD; mean [+/- SD] FEV(1), 88 +/- 5% predicted) were studied as a control group. METHODS: H(2)O(2) and 8-isoprostane levels were measured in EBC, and the values were correlated with variables expressing COPD severity (ie, FEV(1) percent predicted, dyspnea severity score (ie, Medical Research Council scale) and airway inflammation (ie, differential cell counts from induced sputum). RESULTS: The mean concentration of H(2)O(2) was significantly elevated in COPD patients compared to control subjects (mean, 0.66 micromol/L [95% confidence interval (CI), 0.54 to 0.68 micro mol/L) vs 0.31 micro mol/L [95% CI, 0.26 to 0.35 micromol/L], respectively; p < 0.0001). The difference was primarily due to the elevation of H(2)O(2) in patients with severe and moderate COPD, whose expired breath H(2)O(2) levels were significantly higher than those of patients with mild disease (mean, 0.96 micromol/L [95% CI, 0.79 to 1.13 micromol/L], 0.68 micromol/L [95% CI, 0.55 to 0.81 micromol/L], and 0.33 micromol/L [95% CI, 0.24 to 0.43 micromol/L], respectively, p < 0.0001). The mean concentration of 8-isoprostane was significantly elevated in patients with COPD compared to that of the control group (47 pg/mL [95% CI, 41 to 53 pg/mL] vs 29 pg/mL [95% CI, 25 to 33 pg/mL], respectively; p < 0.0001) but did not differ significantly among the different stages of the disease (p = 0.43). Repeatability and stability data within measurements showed that H(2)O(2) has a better repeatability and stability than 8-isoprostane. Furthermore, we observed significant correlations of H(2)O(2) with FEV(1), neutrophil count, and dyspnea score. Those correlations existed only in patients with moderate and severe disease. No correlations were found between levels of 8-isoprostane and the above parameters. CONCLUSIONS: We conclude that levels of H(2)O(2) and 8-isoprostane are elevated in the EBC of patients with COPD, but that H(2)O(2) seems to be a more repeatable and a more sensitive index of the inflammatory process and the severity of the disease.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14555568 !0012-3692 (Print) Journal Article14555568\Pneumonology and Clinical Research Department, Athens Army General Hospital, Athens, Greece. ~?QSandrini, A. Ferreira, I. M. Gutierrez, C. Jardim, J. R. Zamel, N. Chapman, K. R.2003dEffect of montelukast on exhaled nitric oxide and nonvolatile markers of inflammation in mild asthma1334-40Chest1244Acetates/*pharmacology Adult Anti-Asthmatic Agents/*pharmacology Asthma/complications/*drug therapy/*metabolism Biological Markers/analysis Breath Tests Cross-Over Studies Cysteine/*analysis Double-Blind Method Exhalation Female Forced Expiratory Volume/drug effects Humans Hydrogen Peroxide/*analysis Inflammation Mediators/*analysis Leukotrienes/*analysis Male Nitric Oxide/*analysis Peak Expiratory Flow Rate/drug effects Pneumonia/etiology/*metabolism Quinolines/*pharmacology Research Support, Non-U.S. Gov't Severity of Illness IndexOctwSTUDY OBJECTIVES: Leukotriene receptor antagonists appear to exert anti-inflammatory activity in asthma. We undertook the present study to evaluate the effect of montelukast on levels of exhaled nitric oxide (ENO) and two inflammatory markers, hydrogen peroxide (H(2)O(2)), and cysteinyl leukotrienes (cys-LTs), in the exhaled breath condensate of subjects with mild asthma. PATIENTS: Twenty stable subjects with mild asthma (15 women and 5 men; mean [+/- SD] age, 34.8 +/- 12.6 years) were included in the study. INTERVENTION: A 1-week run-in period was followed by 2 weeks of treatment (with montelukast or placebo) that was administered in randomized, double-blind, crossover fashion. One week of washout followed each treatment arm. RESULTS: Montelukast significantly reduced the levels of ENO from baseline (median, 52.5 parts per billion [ppb]; 25th to 75th percentile, 37.8 to 101.8 ppb) during the entire treatment period (ie, day 1 to day 14), with the effect measurable as early as day 1 (median, 45.9 ppb; 25th to 75th percentile, 29.3 to 92.5 ppb) and with the maximal effect being observed on day 7 (median, 35.7 ppb; 25th to 75th percentile, 27.6 to 66.6 ppb). The levels of ENO did not change significantly with placebo therapy. Montelukast improved symptom score and reduced peak expiratory flow (PEF) variability. Changes in PEF variability correlated positively with changes in ENO (r = 0.46; p = 0.04). No significant changes in FEV(1) or concentration of H(2)O(2) in the exhaled breath condensate were observed. Levels of cys-LTs were undetectable in the exhaled breath condensate. CONCLUSIONS: We concluded that montelukast reduces the levels of ENO in patients with mild asthma, a finding that is compatible with an anti-inflammatory effect of montelukast, and that ENO appears to be more sensitive in detecting this effect than FEV(1) and H(2)O(2) levels in the exhaled breath condensate.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14555563 L0012-3692 (Print) Clinical Trial Journal Article Randomized Controlled Trial14555563Asthma & Airway Centre of the Toronto Western Hospital, Division of Respiratory Medicine, University Health Network, University of Toronto, Toronto, ON, Canada. ~?jMoloney, E. D. Mumby, S. E. Gajdocsi, R. Cranshaw, J. H. Kharitonov, S. A. Quinlan, G. J. Griffiths, M. J.2004`Exhaled breath condensate detects markers of pulmonary inflammation after cardiothoracic surgery64-9Am J Respir Crit Care Med1691Aged *Breath Tests Bronchoalveolar Lavage Fluid/chemistry Cohort Studies Comparative Study Coronary Artery Bypass/*adverse effects/methods *Dinoprost/*analogs & derivatives F2-Isoprostanes/analysis Female Humans Inflammation Mediators/*analysis Leukotriene B4/analysis Male Middle Aged Pneumonia/*diagnosis/etiology Postoperative Complications/diagnosis Probability Prognosis Research Support, Non-U.S. Gov't Risk Assessment Sensitivity and Specificity Severity of Illness Index Spectrum Analysis, Mass Statistics, NonparametricJan 11Cardiac 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.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14551168 !1073-449X (Print) Journal Article14551168Unit of Critical Care and Thoracic Medicine, Imperial College London at the National Heart and Lung Institute, Royal Brompton Hospital, London, UK.t~?iEffros, R. M. Biller, J. Foss, B. Hoagland, K. Dunning, M. B. Castillo, D. Bosbous, M. Sun, F. Shaker, R.2003XA simple method for estimating respiratory solute dilution in exhaled breath condensates1500-5Am J Respir Crit Care Med16812Adult Breath Tests/*methods Electric Conductivity Electrolytes/*analysis Extravascular Lung Water/*chemistry Female Humans Male Research Support, U.S. Gov't, P.H.S.Dec 15Exhaled breath condensates have been widely used to detect inflammatory mediators in the fluid that covers airway surfaces of patients with inflammatory lung disorders. This approach is much less invasive than bronchoalveolar lavage, but respiratory droplets are markedly diluted by large and variable amounts of water vapor. We estimated the dilution of respiratory droplets by comparing concentrations of nonvolatile, reference indicators (total nonvolatile cations, urea or conductivity) in 18 normal subjects with normal plasma concentrations by assuming similar concentrations in the respiratory fluid and plasma. The volatile cation, NH4+ (most of which is delivered as NH3 gas from the mouth), represented 93 +/- 3% (SEM) of the condensate cations. More than 99% of the NH4+ was removed by lyophilization, making it possible to use conductivity to estimate total nonvolatile ionic concentrations and facilitating analysis of urea. Conductivity was significantly correlated with electrolyte and urea concentrations. Estimates of dilution based on total cations, conductivity, and urea were not significantly different (cations: 20,472 +/- 2,516; conductivity: 21,019 +/- 2,427; and urea: 18,818 +/- 2,402). These observations suggest that the conductivity of lyophilized samples can be used as an inexpensive, simple, and reliable method for estimating dilution of nonvolatile, hydrophilic mediators in condensates.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=14512268 !1073-449X (Print) Journal Article14512268Division of Pulmonary and Critical Care Medicine, Adult Cystic Fibrosis Center, Medical College of Wisconsin, Milwaukee, WI, USA. effros@mcw.edu|~?Kawamoto, H. Kambe, M.2003-[Effort-independent pulmonary function tests]782-9 Rinsho Byori518Asthma/diagnosis English Abstract Forced Expiratory Flow Rates Humans Pulmonary Disease, Chronic Obstructive/diagnosis Reproducibility of Results Respiratory Function Tests/*methods Sleep Apnea Syndromes/diagnosisAugMany pulmonary function tests require forced expiration, and the precision of the tests depends on the effort of the subjects. In elderly people, the reproducibility of test results may be inadequate because of insufficiency of the subjects' effort, and the diagnosis of COPD, which is frequently observed in elderly people, is often difficult. To improve the accuracy of the diagnosis, pulmonary function tests that do not require effort are needed. In this study, effortless pulmonary function tests (examinations of the cardiogenic oscillation, negative expiratory pressure, exhaled temperature, and exhaled breath condensate), the application of which to various respiratory disorders (i.e., sleep apnea syndrome, COPD, bronchial asthma) is attempted, are presented.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=13677939 (0047-1860 (Print) Journal Article Review13677939|Department of Clinical Laboratory Medicine, Hiroshima University, Graduate School of Biomedical Science, Hiroshima 734-8554. ~?TGessner, C. Hammerschmidt, S. Kuhn, H. Lange, T. Engelmann, L. Schauer, J. Wirtz, H.2003WExhaled breath condensate nitrite and its relation to tidal volume in acute lung injury1046-52Chest1243Acute Disease Adult Aged Biological Markers/analysis *Breath Tests *Critical Care Female Forced Expiratory Volume/physiology Humans Male Middle Aged Nitric Oxide/analysis Nitrites/*analysis Pneumonia/*therapy Predictive Value of Tests Prospective Studies Pulmonary Disease, Chronic Obstructive/*therapy Respiration, Artificial/*adverse effects Respiratory Distress Syndrome, Adult/*diagnosis Risk Factors Tidal Volume/physiologySep)STUDY OBJECTIVE: Mechanical ventilation may damage the lung. Low tidal volume (VT) is protective, but VT is scaled to body weight (BW) and may be high in functionally small ARDS lungs. We hypothesized that exhaled breath condensate (EBC) nitrite (NO(2)(-)) concentration may increase with lung distension. DESIGN: Prospective, noncontrolled study. SETTING: University hospital and medical ICU. PATIENTS: Thirty-five ICU patients requiring mechanical ventilation (severe pneumonia, n = 31; exacerbated COPD, n = 4). Patients were scored according to American and European Consensus Conference on ARDS criteria (AECC) [no lung injury, n = 7; acute lung injury, n = 13; ARDS, n = 15], as well as the Murray lung injury severity score (LISS) [score 0, n = 3; score 0.1 to 2.5, n = 19; score > 2.5, n = 13]. INTERVENTIONS: EBC was collected and analyzed for NO(2)(-), interleukin (IL)-6, and IL-8. Serum was analyzed for IL-6, IL-8, and procalcitonin. RESULTS: and measurements: EBC NO(2)(-) correlated well with VT (milliliters per kilogram of BW; r = 0.79, p < 0.0001) and expiratory minute volume (r = 0.60, p < 0.0001) but not with other ventilatory parameters or parameters of pulmonary (EBC IL-6, EBC IL-8) or systemic (serum IL-6, IL-8, and procalcitonin) inflammation. The ratio of EBC NO(2)(-) and the size of the VT correlated directly with lung injury (AECC, r = 0.66, p < 0.0001; LISS, r = 0.84, p < 0.0001). CONCLUSION: EBC NO(2)(-) increased linearly with VT. The ratio of EBC NO(2)(-) to VT is assumed to reflect NO(2)(-) release at a given VT. An increase in this ratio indicates an inappropriate increase of NO(2)(-) production most likely due to mechanical stress of the remaining open lung units in injured lungs. We conclude that the EBC NO(2)(-)/VT ratio may help to identify situations of critical mechanical stress.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12970036 !0012-3692 (Print) Journal Article12970036vDepartment of Respiratory and Critical Care Medicine, University of Leipzig, Johannisallee 32, 04103 Leipzig, Germany.O~?$Mutti, A. Corradi, M. Rubinstein, I.2003+Reporting data on exhaled breath condensate719; author reply 719Am J Respir Crit Care Med1686Aged Aldehydes/*analysis Asthma/*diagnosis Biological Markers/analysis Breath Tests Child, Preschool Comparative Study Female Humans Male Middle Aged Prognosis Pulmonary Disease, Chronic Obstructive/*diagnosis Sensitivity and Specificity Severity of Illness IndexSep 15fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12963582 1073-449X (Print) Comment Letter12963582~?'Spicuzza, L. Di Maria, G. U. Polosa, R.2003fAdenosine levels in the exhaled breath condensate: a potential surrogate marker of airway inflammation392; author reply 392-3 Eur Respir J222VAdenosine/*analysis Asthma/*metabolism Biological Markers/analysis Breath Tests HumansAugfhttp://www.ncbi.nlm.nih.gov/entrez/queryfcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12952279 0903-1936 (Print) Comment Letter12952279 ~?DVogelberg, C. Hirsch, T. Rosen-Wolff, A. Kerkmann, M. L. Leupold, W.2003Pseudomonas aeruginosa and Burkholderia cepacia cannot be detected by PCR in the breath condensate of patients with cystic fibrosis348-52Pediatr Pulmonol364 Adolescent Adult Breath Tests Burkholderia cepacia/*isolation & purification Child Cystic Fibrosis/*microbiology Humans Polymerase Chain Reaction Pseudomonas Infections/*diagnosis Pseudomonas aeruginosa/*isolation & purification Respiratory Tract Infections/*diagnosis Sputum/microbiologyOctHThe collection of sputum for microbiological examination in young cystic fibrosis patients can be very difficult. However, a knowledge of bacterial flora colonizing the patient's airways is of paramount importance for proper antimicrobial therapy. It is also known that cystic fibrosis patients colonized by Pseudomonas species have a poorer prognosis than Pseudomonas-negative patients. Noninvasive ways of diagnosing airway inflammation that require only minimal cooperation of the patient might yield new possibilities for early detection of airway colonisation. The breath condensate method as a noninvasive diagnostic technique seems especially appropriate for use in children. Therefore, the aim of this study was to evaluate whether the breath condensate method could be used for detection of Pseudomonas species in children with cystic fibrosis. In total, 32 breath condensate and seven sputum samples were obtained from 13 cystic fibrosis patients with Pseudomonas aeruginosa- or Burkholderia cepacia-positive sputum culture (20 samples were obtained during forced expiration). PCR for combined detection of Pseudomonas aeruginosa, Stenotrophomonas maltophilia, and Burkholderia cepacia was performed. PCR results of all breath condensate samples were negative for Pseudomonas aeruginosa, Stenotrophomonas maltophilia, or Burkholderia cepacia, while all sputum sample results were positive. A minimum DNA quantity of 10 fg could be detected in dilution series of the positive control group. We conclude that the breath condensate method cannot be used as a tool for detection of Pseudomonas species.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12950050 !8755-6863 (Print) Journal Article12950050Children's University Hospital, Technical University of Dresden, Fetscherstrasse 74, 01307 Dresden, Germany. christian.vogelberg@mailbox.tu-dresden.de~?JDziedzic, B. Mazanowska-Gajdowicz, J. Walczewska, A. Sarniak, A. Nowak, D.2003pComparison of cadmium and enzyme-catalyzed nitrate reduction for determination of NO2-/NO3- in breath condensate65-74Clin Chim Acta3351-21Adult Biological Markers/analysis Breath Tests/*methods Cadmium/*metabolism Case-Control Studies Catalysis Comparative Study Female Humans Male Middle Aged Nitrate Reductase (NAD(P)H) Nitrate Reductases/*metabolism Nitrates/*analysis Nitrites/*analysis Oxidation-Reduction Research Support, Non-U.S. Gov'tSeplBACKGROUND: Analysis of NO2-/NO3- in expired breath condensate (EBC) has been proposed as a marker of inflammation in various lung diseases. METHODS: NO2- and total NO3-/NO2- concentrations were determined in EBC collected from healthy and asthmatic subjects. The NO3- was first reduced to NO2-, and total NO2- was detected by colorimetric Griess reaction. Two methods of NO3- reduction were compared. To reduce NO3-, cadmium (600 microl EBC-macromethod) and enzyme-NADPH-nitrate reductase (60 microl EBC-micromethod) were used. RESULTS: Macromethod: Mean NO2- concentrations in EBC were 1.64 +/- 0.24 micromol/l in healthy subjects and 0.42 +/- 0.17 micromol/l in asthmatic patients. Mean total NO2-/NO3- levels were 3.64 +/- 0.43 micromol/l in healthy subjects and 3.27 +/- 0.34 micromol/l in asthmatic. Micromethod: NO2- level: 1.69 +/- 0.23 micromol/l in healthy subjects and 0.53 +/- 0.21 micromol/l in asthmatics. Total NO2-/NO3- levels: 3.56 +/- 0.37 micromol/l in healthy subjects and 3.57 +/- 1.17 micromol/l in asthmatics. Variability index was 27% and 6% for macro- and micromethod, respectively. Recovery of NO3- added to EBC was 100% for enzymatic and almost 88% for cadmium reduction. There was no correlation between total NO2-/NO3- levels determined by macro- and micromethod. CONCLUSIONS: We recommend enzymatic reduction as a better method for NO3- determination in EBC.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12927686 40009-8981 (Print) Evaluation Studies Journal Article12927686Department of Experimental and Clinical Physiology, Institute of Biochemistry and Physiology, Medical University of Lodz, Mazowiecka 6/8, Lodz 92-215, Poland. ~?Xvan Beurden, W. J. Harff, G. A. Dekhuijzen, P. N. van der Poel-Smet, S. M. Smeenk, F. W.2003vEffects of inhaled corticosteroids with different lung deposition on exhaled hydrogen peroxide in stable COPD patients242-8 Respiration703Aerosol Propellants Aged Androstadienes/administration & dosage/*pharmacokinetics Beclomethasone/administration & dosage/*pharmacokinetics Breath Tests Bronchodilator Agents/administration & dosage/*pharmacokinetics Cross-Over Studies Female Glucocorticoids/administration & dosage/*pharmacokinetics Humans Hydrocarbons, Fluorinated Hydrogen Peroxide/analysis Male Pulmonary Disease, Chronic Obstructive/*drug therapy Respiratory Function Tests Single-Blind MethodMay-Jun{BACKGROUND: The effects of inhaled corticosteroids (ICS) on markers of oxidative stress in patients with stable COPD are unclear. OBJECTIVES: The aim was to investigate the effect of ICS on exhaled H(2)O(2) in stable COPD patients and to compare ICS with different lung deposition. METHODS: Forty-one stable patients with moderate COPD (FEV(1) approximately 60% predicted) were randomized to sequence 1; first HFA-134a beclomethasone dipropionate (HFA-BDP, an ICS with more peripheral deposition) 400 microg b.i.d., then fluticasone propionate (FP, an ICS with more central deposition) 375 microg b.i.d. (n = 20) or sequence 2; first FP, then HFA-BDP (n = 21). Both 4-week treatment periods were preceded by a 4-week washout period. After each period, the concentration of H(2)O(2) in exhaled breath condensate was measured. RESULTS: The H(2)O(2) concentration decreased significantly after the first treatment period in both sequence 1 and 2 (p < 0.05, p = 0.01, respectively). In neither sequence was there a return to baseline values after the second washout, indicating a carry-over effect. The concentrations remained low in both sequences during the second treatment period. CONCLUSIONS: Both ICS appeared to reduce exhaled H(2)O(2) in stable COPD patients. However, this study showed no difference between ICS with different deposition patterns, which in part may be due to the carry-over effect.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12915742 L0025-7931 (Print) Clinical Trial Journal Article Randomized Controlled Trial12915742bDepartment of Pulmonology, Catharina Hospital Eindhoven, The Netherlands. wendy.beurden@wanadoo.nl~?XCsoma, Z. Bush, A. Wilson, N. M. Donnelly, L. Balint, B. Barnes, P. J. Kharitonov, S. A.2003qNitric oxide metabolites are not reduced in exhaled breath condensate of patients with primary ciliary dyskinesia633-8Chest1242Breath Tests/*methods Case-Control Studies Child Female Humans Kartagener Syndrome/drug therapy/*metabolism Male Nitric Oxide/*metabolism Nitric Oxide Donors/chemistry Research Support, Non-U.S. Gov't S-Nitrosothiols/chemistry Steroids/therapeutic useAugSTUDY OBJECTIVES: To investigate whether nitric oxide (NO) metabolites would be reduced in children affected by primary ciliary dyskinesia (PCD). DESIGN: Single-center observational study. PATIENTS: Fifteen children with PCD (seven boys; mean [+/- SEM] age, 10.3 +/- 0.7 years; mean FEV(1), 73 +/- 2.1% predicted) were recruited along with 14 healthy age-matched subjects (seven boys; mean age, 11.5 +/- 0.4 years; mean FEV(1), 103 +/- 5% predicted). INTERVENTIONS: We assessed the levels of nitrite (NO(2)(-)), NO(2)(-)/NO(3)(-) (NO(2)(-)/NO(3)(-)), and S-nitrosothiol in exhaled breath condensate, exhaled NO, and nasal NO from children with PCD compared to those in healthy children. MEASUREMENTS AND RESULTS: The mean exhaled and nasal NO levels were markedly decreased in children with PCD compared to those without PCD (3.2 +/- 0.2 vs 8.5 +/- 0.9 parts per billion [ppb], respectively [p < 0.0001]; 59.6 +/- 12.2 vs 505.5 +/- 66.8 ppb, respectively [p < 0.001]). Despite the lower levels of exhaled NO in children with PCD, no differences were found in the mean levels of NO(2)(-) (2.9 +/- 0.4 vs 3.5 +/- 0.3 microM, respectively), NO(2)(-)/NO(3)(-) (35.2 +/- 5.0 vs 34.3 +/- 4.5 microM, respectively), or S-nitrosothiol (1.0 +/- 0.2 vs 0.6 +/- 0.1 microM, respectively) between children with PCD and healthy subjects. CONCLUSION: These findings suggest that NO synthase activity may not be decreased as much as might be expected on the basis of low exhaled and nasal NO levels.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12907553 !0012-3692 (Print) Journal Article12907553tDepartment of Thoracic Medicine, Imperial College School of Medicine, National Heart and Lung Institute, London, UK.~?$Makarevich, A. E. Ivashkevich, D. L.2003The importance of exhaled air condensate in assessing the oxidant-antioxidant system in patients with chronic obstructive pulmonary disease19-23Wiad Lek561-2Adult Antioxidants/*metabolism Biological Markers/analysis Breath Tests Case-Control Studies Female Humans *Lipid Peroxidation Male Malondialdehyde/*metabolism Middle Aged Oxidants/*metabolism *Oxidative Stress Pulmonary Disease, Chronic Obstructive/*metabolism/physiopathologyThe aim of this study was to investigate the relationship between selected lipid peroxidation products as markers of oxidative stress and antioxidant defense capacity in exhaled air condensate in comparison to blood and COPD severity. We detected that the increase of lipids peroxidation products in exhaled air condensate (less in blood) was accompanied by the reduction of total antioxidant capacity (antiradical activity and ceruloplasmin) according to the severity of COPD. Thus, the level of malondialdehyde significantly increased both in blood and exhaled air condensate in COPD1,2,3 (by 24%, 86%, 100% and 58%, 92%, 2,3 times respectively). Antiradical activity level was significantly decreased both in blood and exhaled air condensate in patients with COPD2,3 by 24%, 51% and 23.31% respectively. The results of this study clearly demonstrated the imbalance in oxidant-antioxidant system in advanced COPD.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12901263 !0043-5147 (Print) Journal Article12901263\Department of Internal Diseases N1, Belarusian State Medical University. makae@msmi.minsk.by~?Kawamoto, H. Kambe, M.2003[Pulmonary function tests]568-73 Rinsho Byori516REnglish Abstract Humans Respiration Respiratory Function Tests/*methods SpirometryJunThe spirometry has been most valuable pulmonary function test and it defines pulmonary physiology. But the spirometry has not been widely used by general physicians in Japan. The spirometry is effort-dependent test, so, they seem to keep it at a distance. It's desirable that pulmonary function tests are effort-independent. We introduce some effort-independent pulmonary function tests and refer to analysis of exhaled breath condensate.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12884745 (0047-1860 (Print) Journal Article Review12884745|Department of Clinical Laboratory Medicine, Hiroshima University, Graduate School of Biomedical Science, Hiroshima 734-8554.~? Horvath, I.2003;Exhaled breath condensate contains more than only volatiles187-8; author reply 188 Eur Respir J221b*Asthma *Biological Markers Humans *Inflammation Isoprostanes *Lipid Peroxidation Oxidative StressJulfhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12882472 0903-1936 (Print) Comment Letter12882472 <~?WBaraldi, E. Ghiro, L. Piovan, V. Carraro, S. Ciabattoni, G. Barnes, P. J. Montuschi, P.20033Increased exhaled 8-isoprostane in childhood asthma25-31Chest1241Adolescent Anti-Inflammatory Agents/therapeutic use Asthma/diagnosis/drug therapy/*metabolism Breath Tests Chemiluminescent Measurements Child Child, Preschool Cross-Sectional Studies *Dinoprost/*analogs & derivatives Dinoprostone/metabolism F2-Isoprostanes/*metabolism Female Forced Expiratory Volume Humans Male Nitric Oxide/metabolism *Oxidative Stress Radioimmunoassay Respiratory Function Tests SteroidsJulSTUDY OBJECTIVE: To quantify lung oxidative stress in asthmatic children by measuring concentrations of 8-isoprostane, a marker of oxidative stress, in exhaled breath condensate (EBC), which is a noninvasive method of sampling airway secretions. Secondary objectives were as follows: (1) to measure levels of exhaled prostaglandin (PG) E(2), since impaired PGE(2) production has been implicated in the pathogenesis of asthma in adults; and (2) to measure levels of fractional exhaled nitric oxide (FeNO), which is a marker of airway inflammation. DESIGN: Single-center, cross-sectional study. PATIENTS: Twelve healthy children, 12 steroid-naive asthmatic children, and 30 children in stable condition with mild-to-moderate persistent asthma who were being treated with inhaled corticosteroids (ICSs) [average dose, 300 micro g per day] were studied. INTERVENTIONS: Subjects attended the outpatient clinic on one occasion for the collection of EBC and FeNO measurements. Measurements and results: 8-Isoprostane and PGE(2) concentrations in EBC were measured with specific radioimmunoassays. FeNO was measured online by a chemiluminescence analyzer. 8-Isoprostane was detectable in the EBC of healthy children (mean [+/- SEM], 34.2 +/- 4.5 pg/mL), and its concentrations were increased in both steroid-naive asthmatic children (mean, 56.4 +/- 7.7 pg/mL; p < 0.01) and steroid-treated asthmatic children (mean, 47.2 +/- 2.3 pg/mL; p < 0.05). There was no difference in exhaled 8-isoprostane concentrations between the two groups of asthmatic children (p = 0.14). By contrast, exhaled PGE(2) concentrations were similar among the three study groups (p = 0.56). FeNO levels were higher in steroid-naive children with asthma (49.2 +/- 9.6 parts per billion [ppb]; p < 0.05) and, to a lesser extent, in steroid-treated asthmatic children (37.8 +/- 6.6 ppb; p < 0.05) compared with healthy children (15.2 +/- 1.7 ppb). CONCLUSIONS: Lung oxidative stress is increased in children who are in stable condition with asthma, as reflected by increased exhaled 8-isoprostane concentrations. This increase seems to be relatively resistant to treatment with ICSs. Decreased PGE(2) lung production is unlikely to play a pathophysiologic role in childhood asthma.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12853498 !0012-3692 (Print) Journal Article12853498}Department of Pediatrics (Drs. Baraldi, Ghiro, Piovan, and Carraro), School of Medicine, University of Padova, Padova, Italy.>~?:Surovenko, T. N. Iashchuk, A. V. Iansons, TIa Ezhov, S. N.2003P[Efficiency of kinesi- and hydrokinesitherapy in children with bronchial asthma]29-32$Vopr Kurortol Fizioter Lech Fiz Kult30Adolescent Asthma/metabolism/physiopathology/*therapy Breath Tests Breathing Exercises Child Child, Preschool English Abstract *Exercise Therapy Female Humans *Hydrotherapy Kinesiology, Applied/*methods Male Nitric Oxide/metabolism Peak Expiratory Flow Rate/physiology Quality of Life Swimming/physiologyMay-Jun3The authors review efficiency of various programs of kinesi and hydrokinesitherapy of children with atopic bronchial asthma (BA). Efficiency of the treatment was assessed by quality of life using the questionnaire by A. West, D. French "Childhood asthma questionnaire" (adapted for Russia by V. I. Petrov et al). Monitoring of the activity of allergic inflammation of the upper respiratory tracts was performed by examination of the nasal lavage fluid for nitric oxide metabolites, of the lower respiratory tracts--by the metabolites in the condensate of the expired air. It is shown that hydrokinesitherapy raises BA children's quality of life and declines inflammation activity leading to reduction of the number of BA exacerbations and hospitalizations. The above criteria of the treatment efficacy proved sensitive.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12852012 !0042-8787 (Print) Journal Article12852012[Effektivnost' kinezi- i gidrokineziterapii v lechenii detei, bol'nykh bronkhial'noi astmoi. ~?<Montuschi, P. Kharitonov, S. A. Ciabattoni, G. Barnes, P. J.2003/Exhaled leukotrienes and prostaglandins in COPD585-8Thorax587 Administration, Inhalation Adrenal Cortex Hormones/administration & dosage Aged Breath Tests Cross-Sectional Studies Dinoprost/metabolism Dinoprostone/metabolism Female Forced Expiratory Volume/physiology Humans Leukotriene B4/metabolism Leukotriene E4/metabolism Leukotrienes/*metabolism Male Middle Aged Prostaglandin D2/metabolism Prostaglandins/*metabolism Pulmonary Disease, Chronic Obstructive/drug therapy/*metabolism/physiopathology Research Support, Non-U.S. Gov't Thromboxane B2/metabolism Vital Capacity/physiologyJulBACKGROUND: The role of eicosanoids, including leukotrienes (LTs) and prostaglandins (PGs), in chronic obstructive pulmonary disease (COPD) is uncertain. The aim of this study was to investigate whether eicosanoids are measurable in exhaled breath condensate (EBC), a non-invasive method of collecting airway secretions, in patients with stable mild to moderate COPD, and to show possible differences in their concentrations compared with control subjects. METHODS: LTB(4), LTE(4), PGE(2), PGD(2)-methoxime, PGF(2alpha), and thromboxane B(2) (TxB(2)) were measured in EBC in 15 healthy ex-smokers, 20 steroid naive patients with COPD who were ex-smokers, and in 25 patients with COPD who were ex-smokers and who were treated with inhaled corticosteroids. The study was of cross sectional design and all subjects were matched for age and smoking habit. RESULTS: LTB(4) and PGE(2) concentrations were increased in steroid naive (LTB(4): median 100.6 (range 73.5-145.0) pg/ml, p<0.001; PGE(2): 98.0 (range 57.0-128.4) pg/ml, p<0.001) and steroid treated patients with COPD (LTB(4): 99.0 (range 57.9-170.5) pg/ml, p<0.001; PGE(2): 93.6 (range 52.8-157.0) pg/ml, p<0.001) compared with control subjects (LTB(4): 38.1 (range 31.2-53.6) pg/ml; PGE(2): 44.3 (range 30.2-52.1) pg/ml). Both groups of patients had similar concentrations of exhaled LTB(4) (p=0.43) and PGE(2) (p=0.59). When measurable, LTE(4) and PGD(2)-methoxime concentrations were similar in COPD patients and controls, whereas PGF(2alpha) concentrations were increased in the former. TxB(2)-LI was undetectable in any of the subjects. CONCLUSIONS: There is a selective increase in exhaled LTB(4) and PGE(2) in patients with COPD which may be relatively resistant to inhaled corticosteroid therapy.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12832671 !0040-6376 (Print) Journal Article12832671~Department of Pharmacology, School of Medicine, Catholic University of the Sacred Heart, Rome, Italy. pmontuschi@rm.unicatt.it ~?;Szkudlarek, U. Maria, L. Kasielski, M. Kaucka, S. Nowak, D.2003xExhaled hydrogen peroxide correlates with the release of reactive oxygen species by blood phagocytes in healthy subjects718-25 Respir Med976Adult Breath Tests Female Humans Hydrogen Peroxide/*analysis Luminescent Measurements Male Phagocytes/*metabolism Reactive Oxygen Species/blood/*metabolism Research Support, Non-U.S. Gov't Thiobarbituric Acid Reactive Substances/analysisJunVarious cells including polymorphonuclear leukocytes, alveolar macrophages and type-II pneumocytes may be a source of exhaled hydrogen peroxide (H2O2) in airways of humans. H2O2 can convert into hydroxyl radicals leading to peroxidative damage of airways structures and formation of volatile thiobarbituric acid-reactive substances (TBARs). We tested whether exhalation of H2O2 and TBARs by healthy subjects depends on reactive oxygen species generation from blood phagocytes. The expired breath condensate (EBC) and blood specimens were collected from 41 healthy, never smoked subjects (mean age 20.7 +/- 0.8 years, 18 men, 23 women) and then the EBC concentration of H2O2 and TBARs and 2 x 10(-5) M fMLP-provoked whole blood chemiluminescence response was measured. The mean concentration of H2O2 and TBARs in EBC was 0.28 +/- 0.17 and 0.04 +/- 0.13 microM with ratio of positive readings reaching 36/41 and 4/41, respectively. The chemiluminescence response to n-formyl-methionyl-leveyl-phenylalanine stimulation was obtained in all cases and the following parameters were estimated: basal chemiluminescence (bCl); peak chemiluminescence (pCl); absolute light emission (aCl); and peaktime. H2O2 levels in EBC positively correlated (Spearmann test) with bCl (r=0.41, P<0.01), pCl (r=0.47, P<0.01), aCl (r=0.49, P<0.001), peaktime (r=0.52, P<0.001) in the whole group and with bCl (r=0.56, P<0.01), pCl (r=0.67, P<0.01), aCl (r=0.66, P<0.01) in men and with aCl (r=0.41, P<0.05) and peaktime (r=0.48, P<0.05) in women. No association between exhaled TBARs and blood phagocytes activity was found. These results indicate that H2O2 exhalation in healthy never smoked subjects depends on ability of blood phagocytes to generate reactive oxygen species.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12814160 !0954-6111 (Print) Journal Article12814160Department of Experimental and Clinical Physiology, Institute of Physiology and Biochemistry, Medical University of Lodz, Mazowiecka str 6/8, 92-215 Lodz, Poland.u~? Fozard, J. R.2003?The case for a role for adenosine in asthma: almost convincing?264-9Curr Opin Pharmacol339Adenosine/*physiology Adenosine Deaminase/deficiency Animals Asthma/drug therapy/*metabolism Asthma, Exercise-Induced/drug therapy/metabolism Bronchial Hyperreactivity/metabolism Bronchodilator Agents/pharmacology/therapeutic use Humans Receptors, Adenosine A2/physiology Theophylline/pharmacology/therapeutic useJunMice rendered adenosine deaminase-deficient manifest an 'asthma' phenotype in the lungs that includes mast cell degranulation, eosinophilia, mucus hypersecretion and bronchial hyperresponsiveness. These changes can be reversed by enzyme therapy with adenosine deaminase, and attenuated by theophylline. Theophylline also blocks the pro-inflammatory effects of adenosine in allergen-challenged mice. Adenosine A(2A) receptors are an essential part of the physiological negative feedback mechanism for limitation and termination of both tissue-specific and systemic inflammatory responses. In recent clinical studies, increases in plasma adenosine have been shown to accompany exercise-induced asthma, and adenosine concentrations in exhaled breath condensate are increased in asthmatics. These new data provide support for a key role for adenosine in asthma, which has become increasingly persuasive in recent years. The evidence is now convincing, and the time has come for the asthma community to give its full support to the design and evaluation of molecules that mimic or block the biological effects of adenosine as potential novel therapeutics for this condition.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12810190 (1471-4892 (Print) Journal Article Review12810190sResearch Department, Novartis Pharma AG, WSJ 386.508, CH-4002 Basel, Switzerland. john_r.fozard@pharma.novartis.com k~?.Wilhelm, J. Vankova, M. Maxova, H. Siskova, A.2003Hydrogen peroxide production by alveolar macrophages is increased and its concentration is elevated in the breath of rats exposed to hypoxia: relationship to lung lipid peroxidation327-32 Physiol Res523Aldehydes/analysis Analysis of Variance Animals Anoxia/*physiopathology Breath Tests/methods Data Interpretation, Statistical Hydrogen Peroxide/analysis/*metabolism Lipid Peroxidation Lung/metabolism/*physiopathology Macrophages, Alveolar/drug effects/*metabolism/secretion Male N-Formylmethionine Leucyl-Phenylalanine/pharmacology Rats Rats, Wistar Research Support, Non-U.S. Gov't Respiration Tetradecanoylphorbol Acetate/pharmacology Time FactorsXHypoxic exposure triggers a generation of reactive oxygen species that initiate free radical damage to the lung. Hydrogen peroxide is the product of alveolar macrophages detectable in the expired breath. We evaluated the significance of breath H(2)O(2) concentration for the assessment of lung damage after hypoxic exposure and during posthypoxic period. Adult male rats were exposed to normobaric hypoxia (10 % O(2)) for 3 hours or 5 days. Immediately after the hypoxic exposure and then after 7 days or 14 days of air breathing, H(2)O(2) was determined in the breath condensate and in isolated lung macrophages. Lipid peroxidation was measured in lung homogenates. Three-hour hypoxia did not cause immediate increase in the breath H(2)O(2); 5-day hypoxia increased breath H(2)O(2) level to 458 %. After 7 days of subsequent air breathing H2O2 was elevated in both groups exposed to hypoxia. Increased production of H(2)O(2) by macrophages was observed after 5 days of hypoxia and during the 7 days of subsequent air breathing. Lipid peroxidation increased in the periods of enhanced H(2)O(2) generation by macrophages. As the major increase (1040 %) in the breath H(2)O(2) concentration found 7 days after 3 hours of hypoxia was not accompanied by lipid peroxidation, it can be concluded that the breath H(2)O(2) is not a reliable indicator of lung oxidative damage.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12790764 !0862-8408 (Print) Journal Article12790764Department of Medical Chemistry and Biochemistry, Second Faculty of Medicine, Charles University, Prague, Czech Republic. jiri.wilhelm@lfmotol.cuni.cz w~?mBaraldi, E. Carraro, S. Alinovi, R. Pesci, A. Ghiro, L. Bodini, A. Piacentini, G. Zacchello, F. Zanconato, S.2003kCysteinyl leukotrienes and 8-isoprostane in exhaled breath condensate of children with asthma exacerbations505-9Thorax586Adolescent Asthma/diagnosis/*drug therapy/physiopathology Biological Markers/analysis Breath Tests Child Cysteine/*metabolism *Dinoprost/*analogs & derivatives F2-Isoprostanes/*metabolism Female Forced Expiratory Volume/physiology Glucocorticoids/therapeutic use Humans Leukotrienes/*metabolism Male Nitric Oxide/metabolism Peak Expiratory Flow Rate/physiology Prednisone/therapeutic useJunBACKGROUND: Cysteinyl leukotrienes (Cys-LTs) and isoprostanes are inflammatory metabolites derived from arachidonic acid whose levels are increased in the airways of asthmatic patients. Isoprostanes are relatively stable and specific for lipid peroxidation, which makes them potentially reliable biomarkers for oxidative stress. A study was undertaken to evaluate the effect of a course of oral steroids on Cys-LT and 8-isoprostane levels in exhaled breath condensate of children with an asthma exacerbation. METHODS: Exhaled breath condensate was collected and fractional exhaled nitric oxide (FE(NO)) and spirometric parameters were measured before and after a 5 day course of oral prednisone (1 mg/kg/day) in 15 asthmatic children with an asthma exacerbation. Cys-LT and 8-isoprostane concentrations were measured using an enzyme immunoassay. FE(NO) was measured using a chemiluminescence analyser. Exhaled breath condensate was also collected from 10 healthy children. RESULTS: Before prednisone treatment both Cys-LT and 8-isoprostane concentrations were higher in asthmatic subjects (Cys-LTs, 12.7 pg/ml (IQR 5.4-15.6); 8-isoprostane, 12.0 pg/ml (9.4-29.5)) than in healthy children (Cys-LTs, 4.3 pg/ml (2.0-5.7), p=0.002; 8-isoprostane, 2.6 pg/ml (2.1-3.0), p<0.001). After prednisone treatment there was a significant decrease in both Cys-LT (5.2 pg/ml (3.9-8.8), p=0.005) and 8-isoprostane (8.4 pg/ml (5.4-11.6), p=0.04) concentrations, but 8-isoprostane levels remained higher than in controls (p<0.001). FE(NO) levels, which fell significantly after prednisone treatment (p<0.001), did not correlate significantly with either Cys-LT or 8-isoprostane concentrations. CONCLUSION: After a 5 day course of oral prednisone there is a reduction in Cys-LT and 8-isoprostane levels in EBC of children with an asthma exacerbation, although 8-isoprostane levels remain higher than in controls. This finding suggests that corticosteroids may not be fully effective in reducing oxidative stress in children with an exacerbation of asthma.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12775861 !0040-6376 (Print) Journal Article12775861NDepartment of Pediatrics, University of Padova, Italy. eugi@pediatria.unipd.it ~?cCarpagnano, G. E. Kharitonov, S. A. Foschino-Barbaro, M. P. Resta, O. Gramiccioni, E. Barnes, P. J.2003TIncreased inflammatory markers in the exhaled breath condensate of cigarette smokers589-93 Eur Respir J214Adult Breath Tests Carbon Monoxide/metabolism Female Humans Interleukin-6/*metabolism Leukotriene B4/*metabolism Male Smoking/immunology/*metabolism Statistics, NonparametricAprCigarette smoking induces an inflammatory response in the airways that may play a key role in the pathogenesis of chronic obstructive pulmonary disease. Noninvasive markers of inflammation may, therefore, be useful in monitoring the airways of smokers as well as in the screening of subjects at high risk of developing airway obstruction. The aim of the present study was to determine whether the concentrations of the pro-inflammatory cytokine, interleukin (IL)-6, is increased in the exhaled breath condensate of smokers and whether the number of cigarettes smoked has any influence on the exhaled concentrations. The possibility that exhaled IL-6 levels are related to exhaled carbon monoxide (CO) and lung function has also been explored. Another inflammatory marker, leukotriene (LT), was also measured. Twenty-one smokers (39+/-7 yrs, 13 male) and 14 nonsmokers (45+/-6 yrs, eight male) were recruited. IL-6 and LTB4 levels in the breath condensate were measured with an immunoassay kit and exhaled CO examined by means of a modified electrochemical sensor. Higher IL-6 and exhaled CO concentrations were found in current smokers (5.6+/-1.4 pg x mL(-1) and 16.7+/-5.5 parts per million (ppm)) than in nonsmokers (2.6+/-0.2 pg x mL(-1) and 2.1+/-0.6 ppm). Elevated concentrations of LTB4 were also observed in smokers compared to nonsmokers (9.4+/-0.4 pg x mL(-1) versus 6.1+/-0.3 pg x mL(-1)). In addition, there was a correlation between IL-6 concentrations, the number of cigarettes smoked per day, exhaled CO, LTB4 and lung function. Exhaled interleukin-6 and leukotriene B4 levels may be useful noninvasive markers of airway inflammation in cigarette smokers.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12762340 !0903-1936 (Print) Journal Article12762340<Institute of Respiratory Disease, University of Bari, Italy. P~?\Carpagnano, G. E. Kharitonov, S. A. Wells, A. U. Pantelidis, P. Du Bois, R. M. Barnes, P. J.2003gIncreased vitronectin and endothelin-1 in the breath condensate of patients with fibrosing lung disease154-60 Respiration702Biological Markers/analysis Breath Tests Endothelin-1/*analysis Female Humans Immunoenzyme Techniques Lung Diseases, Interstitial/*metabolism Male Middle Aged Pulmonary Fibrosis/*metabolism Vitronectin/*analysisMar-AprBACKGROUND: Non-specific interstitial pneumonia (NSIP) and fibrosing alveolitis associated with systemic sclerosis (FASSc) are diseases of unknown aetiology that are characterised by the accumulation of mononuclear cells, followed by the progressive deposition of collagen within the interstitium and subsequent destruction of lung airspace. Better understanding of mediators involved in fibrosis may be useful for early diagnosis and in clinical monitoring of disease progression. OBJECTIVE: The aim of this study was to investigate the presence of two profibrotic markers, the vitronectin and the endothelin-1 (ET-1) in the airways of NSIP and FASSc patients. METHODS: Ten NSIP (6 males, age 57 +/- 2 years) and 15 FASSc (8 males, age 55 +/- 4 years) patients were recruited along with 10 normal subjects (4 male, age 52 +/- 2 years). Vitronectin and ET-1 concentrations were measured in their breath condensate, using a specific enzyme immunoassay. RESULTS: Higher levels of vitronectin and ET-1 were observed in NSIP and FASSc patients [median 92.8 (91.7-93.9) microg/ml; median 8.3 (7.9-9.3) pg/ml] than in control subjects [median 80.3 (89.3-91.4) microg/ml; p < 0.01; median 5.3 (4.9-5.9) pg/ml, p < 0.0001]. We also found increased concentrations of vitronectin in patients with clinical deterioration compared to those remaining stable and in ex-smokers compared to non-smokers and, increased vitronectin and ET-1 in patients treated with steroids compared to untreated patients. CONCLUSION: These findings justify further studies of vitronectin and ET-1 levels in exhaled breath condensate, as a means of monitoring activity and predicting progression of pulmonary fibrosis.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12740512 !0025-7931 (Print) Journal Article12740512CInstitute of Respiratory Diseases, University of Bari, Bari, Italy.L~? Risby, T. H.20030Further discussion on breath condensate analysis1301-2Am J Respir Crit Care Med16710"Aldehydes/*analysis Biological Markers/analysis Breath Tests/*methods Controlled Clinical Trials Female Humans Lipid Peroxidation Male Pulmonary Disease, Chronic Obstructive/*diagnosis Respiratory Function Tests Sensitivity and Specificity Severity of Illness Index Smoking/*adverse effectsMay 15fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12738595 #1073-449X (Print) Comment Editorial12738595z~?MCarpagnano, G. E. Barnes, P. J. Geddes, D. M. Hodson, M. E. Kharitonov, S. A.2003ZIncreased leukotriene B4 and interleukin-6 in exhaled breath condensate in cystic fibrosis1109-12Am J Respir Crit Care Med1678Adult Breath Tests Cystic Fibrosis/*metabolism Female Humans Interleukin-6/*analysis/metabolism Leukotriene B4/*analysis/metabolism Male Nitric Oxide/analysis Research Support, Non-U.S. Gov'tApr 15Chronic 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, respectively) and an increase in 15 CF patients infected with Pseudomonas aeruginosa (34.3 +/- 5.0 and 9.3 +/- 0.3 pg/m) compared with those infected with other bacteria (18.3 +/- 0.7 and 6.9 +/- 0.5 pg/ml). These findings suggest that LTB4 and IL-6 levels are increased in exhaled breath condensate of patients with CF during exacerbation and could be used to monitor airway inflammation in these patients.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12684249 !1073-449X (Print) Journal Article12684249Department of Thoracic Medicine, National Heart and Lung Institute, Faculty of Medicine, Imperial College London, United Kingdom. ~?0Biernacki, W. A. Kharitonov, S. A. Barnes, P. J.2003nIncreased leukotriene B4 and 8-isoprostane in exhaled breath condensate of patients with exacerbations of COPD294-8Thorax584Adrenal Cortex Hormones/therapeutic use Adrenergic beta-Agonists/therapeutic use Aged Anti-Bacterial Agents/therapeutic use Biological Markers/analysis Breath Tests *Dinoprost/*analogs & derivatives F2-Isoprostanes/*analysis Female Forced Expiratory Volume/physiology Humans Leukotriene B4/*analysis Male Oxidative Stress/physiology Pneumonia/metabolism Pulmonary Disease, Chronic Obstructive/drug therapy/*metabolism Receptors, Adrenergic, beta-2/antagonists & inhibitors Vital Capacity/physiologyAprBACKGROUND: Exacerbations are an important feature of chronic obstructive pulmonary disease (COPD), accounting for a large proportion of health care costs. They are associated with increased airway inflammation and oxidative stress. METHODS: Concentrations of leukotriene B4 (LTB4), a marker of inflammation, and 8-isoprostane, a marker of oxidative stress, were measured in the exhaled breath condensate of 21 patients (11 M) with COPD during an exacerbation and 2 weeks after treatment with antibiotics. In 12 patients who had no further exacerbations these markers were also measured after 2 months. RESULTS: LTB4 concentrations were raised during the COPD exacerbation (mean (SE) 15.8 (1.1) pg/ml and fell after treatment with antibiotics to 9.9 (0.9) pg/ml (p<0.0001). In 12 patients the level of LTB4 fell further from 10.6 (1.1) pg/ml to 8.5 (0.8) pg/ml (p<0.005) after 2 months. In 12 normal age matched subjects the LTB4 levels were 7.7 (0.5) pg/ml. Concentrations of 8-isoprostane were also increased during the exacerbation (13.0 (0.9) pg/ml) and fell after antibiotic treatment to 9.0 (0.6) pg/ml (p<0.0001). In 12 patients there was a further fall from 9.3 (0.7) pg/ml to 6.0 (0.7) pg/ml (p<0.001) after 2 months compared with normal subjects (6.2 (0.4) pg/ml). CONCLUSIONS: Non-invasive markers of inflammation and oxidative stress are increased during an infective exacerbation of COPD and only slowly recover after treatment with antibiotics.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12668789 !0040-6376 (Print) Journal Article12668789Department of Thoracic Medicine, Imperial College School of Medicine at the National Heart and Lung Institute, Royal Brompton Hospital, London SW3 6LY, UK.~?eMontuschi, P. Ragazzoni, E. Valente, S. Corbo, G. Mondino, C. Ciappi, G. Barnes, P. J. Ciabattoni, G.2003FValidation of leukotriene B4 measurements in exhaled breath condensate69-73 Inflamm Res522*Breath Tests Chromatography, High Pressure Liquid/methods Female Humans Immunoenzyme Techniques/standards Leukotriene B4/*analysis Male Middle Aged Research Support, Non-U.S. Gov't Sensitivity and SpecificityFebOBJECTIVE: 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.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12665124 41023-3830 (Print) Journal Article Validation Studies12665124wDepartment of Pharmacology, Catholic University of the Sacred Heart, L.go F. Vito, 1, Roma, Italy. p.montuschi@ic.ac.uk ~?<Andreoli, R. Manini, P. Corradi, M. Mutti, A. Niessen, W. M.2003Determination of patterns of biologically relevant aldehydes in exhaled breath condensate of healthy subjects by liquid chromatography/atmospheric chemical ionization tandem mass spectrometry637-45Rapid Commun Mass Spectrom177qAir Pressure Aldehydes/*analysis/*chemistry Biological Markers/analysis/chemistry *Breath Tests Chromatography, Liquid Health Humans Hydrazines/analysis/chemistry Molecular Structure Oxidative Stress Ozone Reproducibility of Results Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Sensitivity and Specificity Smoke Spectrum Analysis, Mass/*methodsA method for the simultaneous determination of several classes of aldehydes in exhaled breath condensate (EBC) was developed using liquid chromatography/atmospheric pressure chemical ionization tandem mass spectrometry (LC/APCI-MS/MS). EBC is a biological matrix obtained by a relatively new, simple and noninvasive technique and provides an indirect assessment of pulmonary status. The measurement of aldehydes in EBC represents a biomarker of the effect of oxidative stress caused by smoke, disease, or strong oxidants like ozone. Malondialdehyde (MDA), acrolein, alpha,beta-unsaturated hydroxylated aldehydes [namely 4-hydroxyhexenal (4-HHE) and 4-hydroxynonenal (4-HNE)], and saturated aldehydes (n-hexanal, n-heptanal and n-nonanal) were measured in EBC after derivatization with 2,4-dinitrophenylhydrazine (DNPH). Atmospheric pressure chemical ionization of the analytes was obtained in positive-ion mode for MDA, and in negative-ion mode for acrolein, 4-HHE, 4-HNE, and saturated aldehydes. DNPH derivatives were separated on a C18 column using variable proportions of 20 mM aqueous acetic acid and methanol. Linearity was established over 4-5 orders of magnitude and limits of detection were in the 0.3-1.0 nM range. Intra-day and inter-day precision were in the 1.3-9.9% range for all the compounds. MDA, acrolein and n-alkanals were detectable in all EBC samples, whereas the highly reactive 4-HHE and 4-HNE were found in only a few samples. Statistically significant higher concentrations of MDA, acrolein and n-hexanal were found in EBC from smokers.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12661015 !0951-4198 (Print) Journal Article12661015sIstituto Superiore per la Prevenzione e la Sicurezza del Lavoro, Research Center at the University of Parma, Italy.~?HBaraldi, E. Ghiro, L. Piovan, V. Carraro, S. Zacchello, F. Zanconato, S.2003DSafety and success of exhaled breath condensate collection in asthma358-60Arch Dis Child884Adolescent Age Distribution Asthma/*diagnosis/physiopathology Breath Tests/*methods Child Child, Preschool Cross-Sectional Studies Feasibility Studies Forced Expiratory Volume Humans Nitric Oxide/metabolism Research Support, Non-U.S. Gov'tAprBACKGROUND: Exhaled breath condensate (EBC) is a rapidly expanding area of research to study airway inflammation through the detection of volatile and non-volatile substances in the airways. AIMS: To determine the safety and feasibility of EBC procedure in a group of children with asthma of varying severity. METHODS: In a cross sectional study of children aged 4-17 years, 18 healthy and 91 asthmatic children (69 in stable condition and 22 with asthma exacerbation) underwent the EBC procedure. Outcomes assessed included completion of the procedure, decrease in FEV1, change in fractional exhaled nitric oxide (FE(NO)), and adverse effects. No pretreatment with beta2 agonists was given. All children were able to successfully complete the EBC procedure. RESULTS: Median fall in FEV1 after the procedure was -1% (IQR -3.5, 1.8) in asthmatics and was comparable to that observed in healthy children. In only one asthmatic child did the drop in FEV1 exceed 12%. No significant changes in FE(NO) were observed after EBC. CONCLUSION: This study suggests that EBC is a simple and well tolerated method for evaluating biological samples from the lower airway. The procedure was safe in children with asthma exacerbation, and the success rate was 100% in children aged 4 years and above.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12651772 91468-2044 (Electronic) Evaluation Studies Journal Article12651772wDepartment of Pediatrics, Unit of Allergy and Respiratory Medicine, University of Padua, Italy. eugi@pediatria.unipd.it ~?wLuczynska, M. Szkudlarek, U. Dziankowska-Bartkowiak, B. Waszczykowska, E. Kasielski, M. Sysa-Jedrzejowska, A. Nowak, D.2003LElevated exhalation of hydrogen peroxide in patients with systemic sclerosis274-9Eur J Clin Invest333Breath Tests/methods Female Humans Hydrogen Peroxide/analysis/*metabolism Male Middle Aged Reactive Oxygen Species/metabolism Research Support, Non-U.S. Gov't Scleroderma, Systemic/*metabolismMar"BACKGROUND: Systemic sclerosis is accompanied by an influx of activated phagocytes into distal airways. These cells release H2O2, which may evaporate from the airways surface and be detected in expired breath condensate. We tested whether patients with systemic sclerosis exhale more H2O2 than healthy subjects and whether breath condensate H2O2 levels correlate with some clinical parameters. MATERIAL AND METHODS: H2O2 was measured fluorimetrically in the expired breath condensate of 27 patients (22 women, five men, mean age 49 +/- 13.1 years) with systemic sclerosis and 27 age- and sex- matched healthy controls. RESULTS: Exhaled H2O2 levels were 3.5-fold higher (0.88 +/- 0.62 microM vs. 0.25 +/- 0.17 microM, P < 0.001) in the patients with systemic sclerosis than in the controls. Treatment with cyclophosphamide and/or prednisone (29 +/- 50 months, range 3-168 months) did not significantly decrease H2O2 exhalation (0.78 +/- 0.50 microM, n= 10 vs. 0.94 +/- 0.67 microM, n= 17, P > 0.05). No significant difference was found between patients with limited and diffuse scleroderma (1.03 +/- 0.69 microM, n= 17 vs. 0.63 +/- 0.41 microM, n= 10, P > 0.05). H2O2 levels correlated with disease duration (r = 0.38, P < 0.05) and time from the first Raynaud's episode (r = 0.44, P < 0.05). CONCLUSIONS: Patients with systemic sclerosis exhale more H2O2 than healthy controls, suggesting involvement of reactive oxygen species in disease processes. Lack of significant intergroups differences in H2O2 levels may have resulted from the small number of patients analyzed.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12641548 !0014-2972 (Print) Journal Article12641548Department of Experimental and Clinical Physiology, Institute of Physiology and Biochemistry, Medical University of Lodz, Lodz, Poland. ~?$Agarwal, A. R. Mih, J. George, S. C.2003QExpression of matrix proteins in an in vitro model of airway remodeling in asthma35-42Allergy Asthma Proc241MApoptosis/physiology Asthma/complications/*metabolism Comparative Study Extracellular Matrix Proteins/*biosynthesis Gelatinase B/biosynthesis Humans Isometric Contraction/physiology Pulmonary Fibrosis/complications/metabolism RNA, Messenger/biosynthesis Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Respiratory Mucosa/metabolism/pathology Reverse Transcriptase Polymerase Chain Reaction Solubility Stromelysin 1/biosynthesis Time Factors Tissue Inhibitor of Metalloproteinase-1/biosynthesis Transforming Growth Factor beta/biosynthesis Wound Healing/physiologyJan-FebAcute asthma is characterized by a decrease in the pH of the exhaled breath condensate and bronchoconstriction. These perturbations may injure the epithelium in a chronic, intermittent pattern, leading to subepithelial fibrosis. We used an in vitro three-dimensional model of the bronchial mucosa to elucidate the response to a repeated chemical or physical insult to the epithelium in the postcontraction phase. We used enzyme-linked immunosorbent assay and reverse transcriptase--polymerase chain reaction to assess the production of the following proteins: matrix metalloproteinase (MMP) 3, MMP-9, tissue inhibitor of MMP-1, transforming growth factor beta 1, thrombospondin 1, tenascin, and fibronectin. The presence of the epithelium enhanced the degree of tissue contraction (50.1 +/- 4.4% of original area versus 75.4 +/- 2.3%). In the absence of injury, tenascin, fibronectin, MMP-3, and tissue inhibitor of MMP-1 are actively expressed. However, the chronic chemical wound markedly inhibited the expression of all proteins. We conclude that the epithelium, wound type, and age of the tissue (contracting versus postcontraction) impact the expression of key proteins in an in vitro model of subepithelial fibrosis in asthma.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12635576 !1088-5412 (Print) Journal Article12635576Department of Chemical Engineering and Materials Science, 916 Engineering Tower, University of California, Irvine, Irvine, CA 92697-2575, USA.~?Antczak, A. Gorski, P.2002:Markers of pulmonary diseases in exhaled breath condensate317-23Int J Occup Med Environ Health154nAdult Biological Markers/*analysis *Breath Tests Child Humans Lung Diseases/*diagnosis Oxidative Stress PolandExhaled breath condensate has been more and more extensively used as a novel and non-invasive method to study airway inflammation. It is simple to perform, very well tolerated by patients and no adverse events have been reported so far. Serial measurements can be made with no harmful effects on patients, which is of extreme value in occupational medicine. Exhaled breath condensate has been obtained from both adult and children patients suffering from various pulmonary diseases such as asthma, cystic fibrosis, chronic obstructive pulmonary disease, and interstitial lung diseases. Several markers and mediators are detectable in breath condensate: hydrogen peroxide, thiobarbituric acid-reactive substances, isoprostanes, prostaglandins and leukotrienes. Nitric oxide-related markers have also been studied in the condensate. There is increasing body of evidence that changes in condensate markers reflect local abnormalities of airway lining fluid.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12608619 (1232-1087 (Print) Journal Article Review12608619KDepartment of Pneumology and Allergology, Medical University, Lodz, Poland.p~?ONeubauer, B. Struck, N. Mutzbauer, T. S. Schotte, U. Langfeldt, N. Tetzlaff, K.2002Leukotriene-B4 concentrations in exhaled breath condensate and lung function after thirty minutes of breathing technically dried compressed air93-101Int Marit Health531-4Air Breath Tests/methods Diving Enzyme-Linked Immunosorbent Assay Humans Humidity Inflammation/*diagnosis Leukotriene B4/*metabolism Lung Diseases, Obstructive/*diagnosis Male Occupational Diseases/*diagnosis Reference Values Respiratory Function Tests SpirometryYIn previous studies it had been shown that leukotriene-B4 [LTB4] concentrations in the exhaled breath mirror the inflammatory activity of the airways if the respiratory tract has been exposed to occupational hazards. In diving the respiratory tract is exposed to cold and dry air and the nasopharynx, as the site of breathing-gas warming and humidification, is bypassed. The aim of the present study was to obtain LTB4-concentrations in the exhaled breath and spirometric data of 17 healthy subjects before and after thirty minutes of technically dried air breathing at normobar ambient pressure. The exhaled breath was collected non-invasively, via a permanently cooled expiration tube. The condensate was measured by a standard enzyme immunoassay for LTB4. Lung function values (FVC, FEV1, MEF 25, MEF 50) were simultaneously obtained by spirometry. The measured pre- and post-exposure LTB4- concentrations as well as the lung function values were in the normal range. The present data gave no evidence for any inflammatory activity in the subjects' airways after thirty minutes breathing technically dried air.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12608592 !1641-9251 (Print) Journal Article12608592RGerman Naval Medical Institute, D-24119 Kronshagen. Birger.Neubauer@bug.hamburg.de ~?CSandrini, A. Ferreira, I. M. Jardim, J. R. Zamel, N. Chapman, K. R.2003oEffect of nasal triamcinolone acetonide on lower airway inflammatory markers in patients with allergic rhinitis313-20J Allergy Clin Immunol1112Administration, Intranasal Adolescent Adult Aged Asthma/complications Biological Markers Bronchial Hyperreactivity/drug therapy Double-Blind Method Female Forced Expiratory Volume Humans Hydrogen Peroxide/metabolism Inflammation Mediators/metabolism Male Middle Aged Nitric Oxide/metabolism Research Support, Non-U.S. Gov't Respiration Rhinitis, Allergic, Perennial/complications/*drug therapy/physiopathology Rhinitis, Allergic, Seasonal/complications/*drug therapy/physiopathology Triamcinolone Acetonide/*administration & dosageFebfBACKGROUND: Allergic rhinitis (AR) and asthma are commonly associated, and similar underlying inflammatory processes link both diseases. AR, even in the absence of asthma, is associated with increased levels of exhaled nitric oxide (ENO) and hydrogen peroxide (H(2)O(2)) in exhaled breath condensate, 2 noninvasive markers of lower airway inflammation. OBJECTIVE: We sought to evaluate the effect of treatment with the nasal steroid triamcinolone acetonide on ENO and exhaled H(2)O(2) in subjects with AR. METHODS: We allocated 23 subjects in a randomized, double-blind, parallel-controlled fashion to 4-week treatment with triamcinolone acetonide (220 microg/d) or matching placebo. RESULTS: ENO levels were greater in the subgroup with concomitant asthma (16/23 subjects) and decreased significantly with triamcinolone acetonide treatment in this subgroup of patients in comparison with patients receiving placebo. Breath condensate levels of H(2)O(2) were higher in patients with AR without asthma than in those with asthma but decreased significantly with triamcinolone acetonide treatment in both subgroups. No changes were observed in bronchial hyperresponsiveness, nasal and asthma symptoms, or peak expiratory flow with active treatment or placebo. CONCLUSION: We conclude that treatment of AR with triamcinolone acetonide results in decrease of 2 noninvasive markers of lower airway inflammation, ENO and H(2)O(2), supporting that upper and lower airway inflammation should be seen as a continuum in subjects with AR with and without asthma. ENO might be a more specific marker of the lower airway inflammation present in asthma.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12589351 L0091-6749 (Print) Clinical Trial Journal Article Randomized Controlled Trial12589351Asthma and Airway Centre of the Toronto Western Hospital, University Health Network, Division of Respiratory Medicine, University of Toronto, 399 Bathurst Street, Toronto, Ontario M5T 2S8, Canada.~? Hychka, S. H.2002[Diagnostics and prediction of development of pulmonary complications in acute myocardial infarction (morphological validation of the use of non-invasive investigational methods]21-5 Lik Sprava7`Aged Blood-Air Barrier/physiology Breath Tests Bronchopneumonia/*diagnosis/etiology/metabolism Chromatography, Gas *Diagnostic Techniques, Respiratory System English Abstract Fatty Acids/*analysis Humans Lipids/analysis Middle Aged Myocardial Infarction/*complications/metabolism Prognosis Pulmonary Edema/*diagnosis/etiology/metabolism Sweat/chemistryWith the aid of gaschromatographic techniques a fatty-acid composition was studied of lipids of the expired air condensate and sweat of patients in uncomplicated/complicated course of acute myocardial infarction versus morphological changes in the blood-air barrier. The complicated course of the condition vs uncomplicated one has been found out to be accompanied by a significant increase in the lipid content of the polyunsaturated fatty acids--linoleic and arachidonic acids--in the studied biological objects, which fact can be used in diagnosis and prognostication of development of complications in the acute period of myocardial infarction.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12587297 !1019-5297 (Print) Journal Article12587297Diahnostyka ta prohnozuvannia rozvytku lehenevykh uskladnen' hostroho infarktu miokarda (morfolohichne pidhruntia vykorystannia neinvazybnykh metodiv doslidzhennia).~?QCorradi, M. Pesci, A. Casana, R. Alinovi, R. Goldoni, M. Vettori, M. V. Cuomo, A.2003ONitrate in exhaled breath condensate of patients with different airway diseases26-30 Nitric Oxide81Adult Asthma/metabolism *Breath Tests Case-Control Studies Female Humans Lung Diseases/*metabolism Male Middle Aged Nitrates/*analysis Pneumonia/metabolism Pulmonary Disease, Chronic Obstructive/metabolism Smoking/metabolismFebThere is an increasing interest in the measurement of nitric oxide (NO.) in the airways. NO. is a free radical that reacts rapidly with reactive oxygen species in aqueous solution to form peroxynitrite which can then break down to nitrite (NO(2)(-)) and nitrate (NO(3)(-)). NO(3)(-) is considered a stable oxidative end product of NO. metabolism. The aim of this study was to assay NO(3)(-) in exhaled breath condensate (EBC) of normal nonsmoking and smoking subjects, asthmatics, patients with obstructive pulmonary disease (COPD), and patients with community-acquired pneumonia (CAP). EBC was collected using a glass condenser and samples were assayed for NO(3)(-) by ion chromatography followed by conductivity measurement. NO(3)(-) was detectable in EBC of all subjects. NO(3)(-) was elevated in smokers [median (range)] [62.5 (9.6-158.0) microM] and in asthmatics [68.0 (25.8-194.6) microM] compared to controls [9.6 (2.6-119.4) microM; p=0.003 and p=0.006, respectively], whereas NO(3)(-) was not elevated in COPD patients [24.1 (1.9-337.0 microM]. The concentration of NO(3)(-) in patients with CAP [243.4 (26.1-584.5) microM] was higher than that in controls (p=0.002) and NO(3)(-) values decreased after treatment and recovery from illness [40.0 (4.1-167.0) microM, p=0.009]. This study shows that NO(3)(-) is detectable in EBC of healthy subjects and it varies in patients with inflammatory airway diseases.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12586538 !1089-8603 (Print) Journal Article12586538Dipartimento di Clinica Medica, Nefrologia e Scienze della Prevenzione, Universita degli Studi di Parma, Via Gramsci 14, 43100 Parma, Italy. m.corradi@iol.it\~?%Wood, L. G. Gibson, P. G. Garg, M. L.2003@Biomarkers of lipid peroxidation, airway inflammation and asthma177-86 Eur Respir J211b*Asthma *Biological Markers Humans *Inflammation Isoprostanes *Lipid Peroxidation Oxidative StressJan.Oxidative stress, specifically lipid peroxidation, is believed to contribute to the pathophysiology of asthma. This review highlights the pathways through which reactive oxygen species (ROS) may lead to lipid peroxidation. The potential of both the innate and acquired immune systems to activate inflammatory cells and release ROS that may overwhelm the host antioxidant defences and cause lipid peroxidation, accompanied by detrimental pathophysiological effects, are discussed. Despite the evidence demonstrating the importance of lipid peroxidation, systematic characterisation of oxidative stress and antioxidant defences has not been undertaken, largely due to the lack of appropriate biomarkers. This review discusses the emergence of isoprostanes (specifically 8-iso-prostaglandin F2alpha) as reliable, in vivo markers of lipid peroxidation, which provides an appropriate tool for studying oxidative stress. Furthermore, the development of techniques to study induced sputum and breath condensate, derived directly from the airway surface, enables the site of oxidative damage to be closely assessed. Evidence suggests that dietary changes that have occurred over recent years have increased susceptibility to lipid peroxidation, due to reduced antioxidant defences. To date, the limited number of long-term (>1 week) supplementation trials have been promising. However, the development of techniques to study isoprostanes in airway-lining fluid pave the way for further studies investigating the potential for antioxidant supplements to be used as routine therapy in asthma.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12570126 (0903-1936 (Print) Journal Article Review12570126Hunter Medical Research Institute, Dept of Respiratory and Sleep Medicine, John Hunter Hospital, New Lambton Heights, Australia. ~? Disse, B.2002bClinical evaluation of new therapies for treatment of mucus hypersecretion in respiratory diseases 254-72; discussion 272-6, 277-82Novartis Found Symp248 Anti-Inflammatory Agents/pharmacology/therapeutic use Biological Markers Clinical Trials/methods/standards Drug Evaluation Exocytosis/*drug effects Expectorants/pharmacology/therapeutic use Humans Mucus/chemistry/drug effects/*secretion Predictive Value of Tests Pulmonary Disease, Chronic Obstructive/drug therapy/physiopathology Respiratory Function Tests Respiratory System Agents/pharmacology/*therapeutic use Respiratory Tract Diseases/*drug therapy/physiopathology/therapy Safety Smoking Cessation Treatment Outcome,In the past mucoactive drugs in airway diseases have been identified and profiled in symptom-based animal experiments and in clinical trials along related lines (cough and expectoration). Presently available drugs of this class are not generally accepted by licensing authorities worldwide and no new molecule clinically profiled as a mucoactive drug has been brought to regulatory approval in the past 20 years. Among regulatory guidelines only the CPMP 1999 'points to consider' on drug development in chronic obstructive pulmonary disease (COPD) advises for mucoactive drug development by suggesting that an indication for symptomatic treatment may be established on the basis of a symptom-related primary endpoint that should be justified as for its importance and supported by a co-primary lung function endpoint. Quality and safety of the new drug must be documented in long-term studies and the indication and use clearly described based on established or adequately profiled new primary endpoints in two pivotal studies. Published trials on mucoactive drugs have used a variety of endpoints. These include mucus hypersecretion-related symptoms by questionnaire, expectorated volume and dry weight, and mucus viscosity, elasticity and transportability. Most methods and endpoints are not validated and a positive standard of treatment is not established. New surrogate markers of efficacy for shorter term trials, e.g. induced or spontaneous sputum based assays (cellularity, mucus antigens), exhaled breath (NO), breath condensate (eicosanoids) or airway biopsy are only partially validated and the risk of false positive or negative phase II results is appreciably high. On the other hand, lung function measurements including airway hyper-reactivity assessment and typical phase III (long-term) endpoints like dyspnoea ratings, health status assessments, incidence of exacerbations and lung function decline over time are validated endpoints and offer a high likelihood of regulatory acceptance. Proof for no depression of lung mucociliary clearance is an important safety endpoint.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12568499 (1528-2511 (Print) Journal Article Review12568499zDepartment of Therapeutic Area Respiratory Diseases, Boehringer Ingelheim Pharma KG, PO Box 200, 55216 Ingelheim, Germany.v~? Gaston, B.2003=Breath condensate analysis: perhaps worth studying, after all292-3Am J Respir Crit Care Med1673DAsthma/metabolism Breath Tests/*methods Glutathione/*analysis HumansFeb 1fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12554618 #1073-449X (Print) Comment Editorial125546185~?*Sampson, A. P. Pizzichini, E. Bisgaard, H.2003aEffects of cysteinyl leukotrienes and leukotriene receptor antagonists on markers of inflammationS49-59; discussion S59-61J Allergy Clin Immunol1111 SupplAnti-Asthmatic Agents/*pharmacology/therapeutic use Anti-Inflammatory Agents/*pharmacology/therapeutic use Asthma/*drug therapy/genetics/immunology Biological Markers/analysis/blood/urine Bronchoalveolar Lavage Fluid/chemistry Glutathione Transferase/genetics Humans Inflammation/blood/urine Leukotriene Antagonists/*pharmacology/therapeutic use Nitric Oxide/analysis SRS-A/*physiology Sputum/chemistryJanThe understanding that asthma pathophysiology includes an inflammatory component has spurred the more aggressive use of anti-inflammatory therapies and created a need for effective tools to measure inflammation. Biomarkers of airway inflammation proposed are obtained by methods that are direct but highly invasive (bronchial biopsy, bronchoalveolar lavage), moderately direct, and less invasive (indirect sputum, exhaled air, breath condensate) or indirect and least invasive (blood, urine). Several studies described in this review have implicated the cysteinyl leukotrienes (CysLTs) as inflammatory mediators in a wide range of diseases, implying that their biological activities reach far beyond acute bronchoconstriction, the activity traditionally ascribed to them. The validity of examining sputum for "biomarkers" has improved the understanding of asthma pathophysiology, optimization of asthma treatment and management, and investigation of the relation between CysLTs and airway inflammation in asthma. Nitric oxide is also a surrogate marker of asthma and reflects airway inflammation. The anti-inflammatory effects of the leukotriene receptor antagonists and the markers of their activity continue to grow.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12532086 (0091-6749 (Print) Journal Article Review12532086KRespiratory Cell Division at Southampton General Hospital, Southampton, UK. e~?^Corradi, M. Rubinstein, I. Andreoli, R. Manini, P. Caglieri, A. Poli, D. Alinovi, R. Mutti, A.2003]Aldehydes in exhaled breath condensate of patients with chronic obstructive pulmonary disease1380-6Am J Respir Crit Care Med167108Adult Age Factors Aged Aldehydes/*analysis/metabolism Biological Markers/analysis Breath Tests/*methods Case-Control Studies Cohort Studies Female Humans Lipid Peroxidation/physiology Male Malondialdehyde/analysis Middle Aged Probability Prognosis Pulmonary Disease, Chronic Obstructive/*diagnosis Reference Values Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S. Respiratory Function Tests Risk Assessment Sensitivity and Specificity Severity of Illness Index Sex Factors Smoking/*adverse effects/physiopathology Statistics, NonparametricMay 15The aims of the present study were (1) to evaluate whether individual aldehydes resulting from lipid peroxidation can be measured in exhaled breath condensate, (2) to assess the influence of sampling procedures on aldehyde concentrations, and (3) to compare aldehyde levels of patients with stable, moderate to severe, chronic obstructive pulmonary disease with those of smoking and nonsmoking control subjects. Aldehydes (malondialdehyde, hexanal, heptanal, and nonanal) were measured by liquid chromatography-tandem mass spectrometry in all samples and overlapping results were obtained by different sampling procedures. Malondialdehyde (57.2 +/- 2.4 nmol/L), hexanal (63.5 +/- 4.4 nmol/L), and heptanal (26.6 +/- 3.9 nmol/L) were increased in patients as compared with nonsmoking control subjects (17.7 +/- 5.5 nmol/L, p < 0.0001; 14.2 +/- 3.5 nmol/L, p = 0.004; and 18.7 +/- 0.9 nmol/L, p = 0.002, respectively). Only malondialdehyde was increased in patients compared with smoking control subjects (35.6 +/- 4.0 nmol/L, p = 0.0007). In conclusion, different classes of aldehydes were identified in exhaled breath condensate of humans. Whereas all aldehydes but nonanal were lower in control subjects as compared with other groups, only malondialdehyde distinguished smoking control subjects from patients with chronic obstructive pulmonary disease and could be envisaged as a biomarker potentially useful to monitor the disease and its response to therapy.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12522029 !1073-449X (Print) Journal Article12522029National Institute of Occupational Safety and Prevention Research Center, Laboratory of Industrial Toxicology, Department of Clinical Medicine, Nephrology and Health Sciences, University of Parma, Italy. T~?]Huszar, E. Vass, G. Vizi, E. Csoma, Z. Barat, E. Molnar Vilagos, G. Herjavecz, I. Horvath, I.2002XAdenosine in exhaled breath condensate in healthy volunteers and in patients with asthma1393-8 Eur Respir J206Adenosine/*analysis Adult Asthma/*metabolism Breath Tests Case-Control Studies Female Humans Male Reproducibility of Results Research Support, Non-U.S. Gov't Respiratory Function Tests Saliva/chemistryDec|Persistent airway inflammation may require the use of different markers for monitoring airway inflammation. In this study, the authors investigated whether adenosine, which may be produced in allergic inflammatory conditions, could be measured with good reproducibility in exhaled breath condensate (EBC), and whether its concentration was elevated in patients with asthma. EBC adenosine and exhaled nitric oxide (eNO), a noninvasive marker of asthmatic airway inflammation, were measured in 40 healthy volunteers and 43 patients with allergic bronchial asthma. Repeatability of adenosine measurement was checked in 20 pairs of samples collected from healthy control subjects. Adenosine was detectable in all EBC samples by the applied high-performance liquid chromatographic method. The mean difference between repeated measurements of adenosine was -0.1 nM and all differences were within the coefficient of repeatability. Adenosine concentration was higher in steroid-naive patients (n=23) compared with healthy control subjects and steroid-treated patients (n=20). In patients with worsening symptoms of asthma (n=23), adenosine concentration was elevated compared with those in a stable condition (n=20). Furthermore, adenosine concentrations were related to eNO levels in asthmatic patients. These results, showing good reproducibility of adenosine measurements and increased adenosine concentrations in steroid-naive patients and in patients with worsening of asthmatic symptoms, indicate that adenosine measurement in exhaled breath condensate might be an acceptable novel method to investigate the role of local production of adenosine in the airways.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12503694 !0903-1936 (Print) Journal Article12503694zDept of Pathophysiology, National Koranyi Institute for Tuberculosis and Pulmonology, Budapest, Hungary. huszar@koranyi.hu~? Effros, R. M.2003]Do low exhaled condensate NH4+ concentrations in asthma reflect reduced pulmonary production?91; author reply 91-2Am J Respir Crit Care Med1671aAsthma/*metabolism Breath Tests Humans Lung/*metabolism Quaternary Ammonium Compounds/*metabolismJan 1fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12502482 1073-449X (Print) Comment Letter12502482 ~?[Vass, G. Huszar, E. Barat, E. Valyon, M. Kiss, D. Penzes, I. Augusztinovicz, M. Horvath, I.2003SComparison of nasal and oral inhalation during exhaled breath condensate collection850-5Am J Respir Crit Care Med1676Adenosine/*analysis Adult Ammonia/*analysis Bias (Epidemiology) Breath Tests/instrumentation/*methods Case-Control Studies Comparative Study Constriction Female Forced Expiratory Volume Humans Inflammation Mediators/*analysis *Inhalation Linear Models Male *Mouth *Nose Research Support, Non-U.S. Gov't Rhinitis, Allergic, Perennial/*diagnosis/*physiopathology Specimen Handling/instrumentation/*methods/standards Thromboxane B2/*analysis Tidal Volume Time Factors Vital CapacityMar 15Analysis of exhaled breath condensate is a method for noninvasive assessment of the lung. Condensate can be collected with a nose clip (subjects inhale and exhale via the mouth) or without it (subjects inhale via the nose and exhale via the mouth), but the mode of inhalation may influence condensate volume and mediator levels. We compared condensate volume and adenosine, ammonia, and thromboxane B2 levels in young healthy volunteers (n = 25) in samples collected for 10 minutes from subjects with or without a nose clip. Patients with allergic rhinitis (n = 8) were also studied to assess the effect of upper airway inflammation on mediator levels. Adenosine, ammonia, and thromboxane B2 levels were determined by HPLC, spectrophotometry, and radioimmunoassay, respectively. Volume of condensate was significantly higher without nose clip than that with nose clip (mean +/- SD, 2321 +/- 736 microl and 1746 +/- 400 microl, respectively; p = 0.0001). We found no significant difference in any mediator levels between these two collection modes in healthy volunteers, but adenosine showed a tendency to differ between oral and nasal inhalation in patients with allergic rhinitis. Our data indicate that whereas a greater volume of condensate can be obtained when subjects inhale through their noses, the mode of inhalation does not influence mediator levels in young healthy volunteers, but may affect these levels in patients with allergic rhinitis.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12493648 41073-449X (Print) Journal Article Validation Studies12493648National Koranyi Institute for Pulmonology, Department of Pathophysiology, Budapest, Piheno ut 1., P.O. Box 1 H-1529, Hungary. hildiko@koranyi.hu ~?*Paredi, P. Kharitonov, S. A. Barnes, P. J.2002.Analysis of expired air for oxidation productsS31-7Am J Respir Crit Care Med16612 Pt 2&Asthma/*metabolism Biological Markers/*analysis Breath Tests/*methods Carbon Monoxide/analysis Cystic Fibrosis/*metabolism Humans Hydrocarbons/analysis Lipid Peroxidation/physiology Oxidative Stress/*physiology Pulmonary Disease, Chronic Obstructive/*metabolism Research Support, Non-U.S. Gov'tDec 15Chronic inflammation is a critical feature of chronic obstructive pulmonary disease, cystic fibrosis, and asthma. This inflammation is associated with the increased production of reactive oxygen species or oxidative stress in the lungs. Oxidative stress may have several adverse effects and may amplify the inflammatory process; however, monitoring oxidative stress is difficult and may not be reflected by changes in blood markers. We have therefore developed several noninvasive markers in the exhaled breath that may indicate oxidative stress in the lungs, and we studied these in relationship to the severity of chronic inflammatory lung diseases. We analyzed the exhaled breath for the content of nitric oxide as a marker of inflammation, carbon monoxide as a marker of oxidative stress, and ethane, which is one of the end products of lipid peroxidation. In addition, we measured the concentration of markers of oxidative stress such as isoprostanes in exhaled breath condensate. Our results confirm that there are increased inflammation, oxidative stress, and lipid peroxidation in lung disease, as shown by elevated levels of nitric oxide, carbon monoxide, and ethane, respectively. The finding of lower levels of these gases in patients on steroid treatment and of higher levels in those with more severe lung disease, as assessed by lung function tests and clinical symptoms, reinforces the hypothesis that the noninvasive measurement of exhaled gases maybe useful in monitoring the underlying pathologic pathways of lung disease. Longitudinal studies are required to assess the clinical usefulness of these measurements in the monitoring of chronic inflammatory lung disease.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12471086 (1073-449X (Print) Journal Article Review12471086Department of Thoracic Medicine, National Heart and Lung Institute, Imperial College School of Science, Technology and Medicine, London, United Kingdom.~?@Montuschi, P. Nightingale, J. A. Kharitonov, S. A. Barnes, P. J.2002fOzone-induced increase in exhaled 8-isoprostane in healthy subjects is resistant to inhaled budesonide1403-8Free Radic Biol Med3310Adult Breath Tests Budesonide/*pharmacology Cross-Over Studies Dinoprost/*chemistry Double-Blind Method F2-Isoprostanes/*pharmacology Female Free Radicals Humans Immunoenzyme Techniques Lung/physiology Male *Oxidative Stress Ozone/pharmacology Placebos Protein Isoforms Random Allocation Research Support, Non-U.S. Gov't Smoking Sputum/metabolism Time Factors Vasoconstrictor Agents/pharmacologyNov 15The aim of this study was to quantify lung oxidant stress after short-term ozone exposure as reflected by 8-isoprostane concentrations in exhaled breath condensate (EBC) and to investigate the effects of inhaled budesonide on this response. 8-Isoprostane is a prostaglandin-F(2 alpha) isomer that is formed in vivo by free radical-catalyzed peroxidation of arachidonic acid. EBC is a noninvasive method to collect airway secretions. We undertook a double-blind, randomized, placebo-controlled, crossover study with inhaled budesonide (800 microg) or placebo twice daily for 2 weeks prior to ozone exposure (400 parts per billion) for 2 h in nine healthy nonsmokers. Exhaled 8-isoprostane was measured by an enzyme immunoassay. 8-Isoprostane was increased 4 h after ozone exposure compared to pre-exposure values in both placebo (36.9 +/- 3.9 pg/ml, mean +/- SEM, vs. 16.9 +/- 0.7 pg/ml; p <.001) and budesonide groups (33.4 +/- 2.6 pg/ml vs. 15.8 +/- 0.3 pg/ml; p <.001). Pretreatment with budesonide did not affect the increases in 8-isoprostane (mean differences 3.4 pg/ml, 95% CI -8.9 to 15.7, p =.54). Short-term ozone exposure causes acute increase in lung oxidative stress as reflected by exhaled 8-isoprostane. This increase is resistant to pretreatment with a high dose of inhaled budesonide.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12419472 L0891-5849 (Print) Clinical Trial Journal Article Randomized Controlled Trial12419472zDepartment of Pharmacology, School of Medicine, Catholic University of the Sacred Heart, Rome, Italy. p.montuschi@ic.ac.uk~?pCorradi, M. Folesani, G. Andreoli, R. Manini, P. Bodini, A. Piacentini, G. Carraro, S. Zanconato, S. Baraldi, E.2003[Aldehydes and glutathione in exhaled breath condensate of children with asthma exacerbation395-9Am J Respir Crit Care Med1673Asthma/*metabolism/physiopathology Breath Tests/*methods Child Female Glutathione/*analysis Humans Male Malondialdehyde/*analysisFeb 1Oxidative stress is implicated in the pathogenesis of asthma, and clinical studies show an imbalance in the level of oxidants to the level of antioxidants in subjects with asthma. Aldehydes and glutathione are examples of biomarkers of oxidant-induced damage and antioxidant status in asthma, respectively. In the study, we applied analytical techniques based on liquid chromatography for the assessment of aldehydes and glutathione in the exhaled breath condensate of children with asthma and in control subjects without asthma. Twelve subjects with asthma were evaluated at exacerbation and after 5 days of therapy with prednisone. At exacerbation, malondialdehyde levels were higher in patients with asthma (30.2 +/- 2.4 nM) than in control subjects (19.4 +/- 1.9 nM, p = 0.002) and were reduced after steroid therapy (18.5 +/- 1.6 nM, p = 0.001). At exacerbation, glutathione levels were lower in subjects with asthma (5.96 +/- 0.6 nM) than in control subjects (14.1 +/- 0.8 nM, p < 0.0001) and were increased after the therapy (8.44 +/- 1.2 nM, p = 0.04). Malondialdehyde and glutathione both in subjects with asthma and control subjects were negatively correlated (r = -0.5, p = 0.001). The study shows that aldehydes and glutathione are detectable in the exhaled breath condensate of children with asthma and healthy children and that their levels are modified during asthma exacerbation and after a 5-day course of therapy with oral prednisone.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12411284 !1073-449X (Print) Journal Article12411284Department of Clinical Medicine, Nephrology and Health Sciences, Research Center, National Institute of Occupational Safety and Prevention at the University of Parma, Parma, Italy. corradi@nemo.unipr.it ]~?KCsoma, Z. Kharitonov, S. A. Balint, B. Bush, A. Wilson, N. M. Barnes, P. J.2002GIncreased leukotrienes in exhaled breath condensate in childhood asthma1345-9Am J Respir Crit Care Med16610Adolescent Asthma/drug therapy/*metabolism Biological Markers *Breath Tests Child Child Welfare Comparative Study Cysteine/*metabolism Female Forced Expiratory Volume/physiology Great Britain Humans Leukotriene B4/*metabolism Leukotrienes/*metabolism Male Nitric Oxide/metabolism Research Support, Non-U.S. Gov't *Respiration/drug effects Severity of Illness Index Statistics Steroids, Fluorinated/therapeutic use Treatment OutcomeNov 15}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 and moderate to severe persistent asthma compared with normal subjects (27.9 +/- 2.8 and 31.5 +/- 4.5 versus 18.5 +/- 0.5 pg/ml, p < 0.01 and p < 0.05). There was an inverse correlation between exhaled cys-LTs and LTB4 in patients with mild persistent asthma. We conclude that exhaled cys-LTs and LTB4 may be noninvasive markers of airway inflammation in pediatric asthma.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12406853 !1073-449X (Print) Journal Article12406853Department of Thoracic Medicine and Department of Pediatric Respiratory Care, Imperial College School of Medicine, National Heart and Lung Institute, London, United Kingdom. "~?=Tate, S. MacGregor, G. Davis, M. Innes, J. A. Greening, A. P.2002PAirways in cystic fibrosis are acidified: detection by exhaled breath condensate926-9Thorax5711Adult Breath Tests Carbon Dioxide/*analysis Cystic Fibrosis/*metabolism Female Humans Hydrogen-Ion Concentration Longitudinal Studies Male Nitrites/analysis Pulmonary Alveoli/*chemistry Research Support, Non-U.S. Gov'tNov#BACKGROUND: The loss of cystic fibrosis transmembrane conductance regulator (CFTR) mediated chloride conductance does not fully explain the diverse pathologies evident in patients with cystic fibrosis (CF). Bicarbonate (HCO(3)(-)) secretion is also impaired in CFTR expressing tissues and CFTR is thought to regulate HCO(3)(-) secretion at the apical membrane of epithelial cells. We hypothesised that the epithelial lining fluid (ELF) of patients with CF would be acidified and that this may be worsened during an infective exacerbation due to the increased inflammatory burden. METHODS: pH and nitrite levels in exhaled breath condensate (EBC) from 12 healthy non-smoking controls and 30 patients with CF (11 of whom were in an infective exacerbation) were measured. A further nine patients were studied before and after intravenous antibiotic treatment for an exacerbation of CF. RESULTS: The pH of EBC was significantly lower in patients with stable CF than in controls (5.88 (0.32) v 6.15 (0.16), p=0.017), and was further reduced in CF patients with an exacerbation (5.32 (0.38), p=0.001) compared with stable CF patients. EBC pH increased significantly following antibiotic treatment from 5.27 (0.42) to 5.71 (0.42), p=0.049). Nitrite levels in EBC were increased in CF patients with an exacerbation compared with control subjects (4.4 (4.0) micro m v 1.6 (1.6) micro m p=0.047). No correlation was found between EBC pH and nitrite levels. CONCLUSIONS: These findings support the hypothesis that airway acidification occurs in CF. This acidity is in part a function of inflammation as the pH of the EBC of patients increased significantly with treatment of an exacerbation, although not to control levels. Acidic pH of the ELF may play a role in the pathophysiology of CF lung disease and requires further investigation.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12403872 !0040-6376 (Print) Journal Article12403872Scottish Adult Cystic Fibrosis Service, Respiratory Medicine Unit, Western General Hospital and Medical Genetics Section, University of Edinburgh, Edinburgh, EH4 2XU, UK. steve.tate@ed.ac.uk ~?cCarpagnano, G. E. Kharitonov, S. A. Resta, O. Foschino-Barbaro, M. P. Gramiccioni, E. Barnes, P. J.2002bIncreased 8-isoprostane and interleukin-6 in breath condensate of obstructive sleep apnea patients1162-7Chest1224Adult Biological Markers/analysis Blood Gas Analysis Breath Tests/methods Case-Control Studies Comparative Study Female Humans Inflammation Mediators/analysis Interleukin-6/*analysis Isoprostanes/*analysis Male Middle Aged Obesity/diagnosis/physiopathology Oxidative Stress Polysomnography Probability Prognosis Prospective Studies Reference Values Respiratory Function Tests Sensitivity and Specificity Severity of Illness Index Sleep Apnea, Obstructive/*diagnosis/physiopathology Statistics, NonparametricOctSTUDY OBJECTIVES: Obstructive sleep apnea (OSA) is characterized by repeated episodes of upper airways obstruction during sleep that result in episodes of hypoxia. An increase of systemic biomarkers of inflammation and oxidative stress has been found in patients with OSA and obesity. DESIGN: The aim of this study was to measure the levels of markers of inflammation (interleukin [IL]-6) and oxidative stress (8-isoprostane) in the exhaled breath condensate of OSA and obese patients. PATIENTS AND METHODS: Eighteen OSA patients (13 men; mean [+/- SEM] age, 44 +/- 7 years), 10 obese subjects (4 men; mean age, 39 +/- 8 years), and 15 healthy age-matched subjects (8 men; mean age, 42 +/- 4 years) were recruited. IL-6 and 8-isoprostane were measured in exhaled breath condensate by a specific enzyme immunoassay kit. Measurements and results: Higher concentrations of IL-6 were found in OSA patients (8.7 +/- 0.3 pg/mL) than in healthy control subjects (1.6 +/- 0.1 pg/mL; p < 0.0001). Obese subjects also had higher levels than healthy control subjects, but lower levels than OSA patients (2.1 +/- 0.2 pg/mL, p < 0.05 and p < 0.0001 respectively). Furthermore, 8-isoprostane levels were found to be higher in OSA patients (7.4 +/- 0.7 pg/mL) than in obese subjects (5 +/- 0.3 pg/mL; p = 0.4) and healthy subjects (4.5 +/- 0.5 pg/mL; p < 0.005). We found a positive correlation between these two markers and neck circumference and apnea/hypopnea index. CONCLUSIONS: These findings suggest that inflammation and oxidative stress are characteristic in the airways of OSA patients but not in obese subjects, and that their levels depend on the severity of the OSA. The measurement of IL-6 and 8-isoprostane levels may prove to be useful in screening and monitoring obese patients who have a high risk of developing OSA.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12377837 !0012-3692 (Print) Journal Article12377837CInstitute of Respiratory Diseases, University of Bari, Bari, Italy.~?XEmelyanov, A. Fedoseev, G. Krasnoschekova, O. Abulimity, A. Trendeleva, T. Barnes, P. J.2002fTreatment of asthma with lipid extract of New Zealand green-lipped mussel: a randomised clinical trial596-600 Eur Respir J203Adolescent Adult Animals Anti-Asthmatic Agents/therapeutic use Anti-Inflammatory Agents, Non-Steroidal/*therapeutic use Asthma/*drug therapy/physiopathology Bivalvia Breath Tests Double-Blind Method Forced Expiratory Volume Humans Hydrogen Peroxide/analysis Lipids/*therapeutic use Lipoxygenase Inhibitors/*therapeutic use Middle Aged Peak Expiratory Flow Rate Tissue Extracts/therapeutic useSep"Asthma is a chronic inflammatory disease of the airways mediated, at least in part, by leukotrienes and other lipid mediators. Experimental studies have shown that lipid extract of New Zealand green-lipped mussel, Perna canaliculus, is effective in inhibiting 5'-lipoxygenase and cyclo-oxygenase pathways responsible for production of eicosanoids, including leukotrienes and prostaglandins. The aim of this study was to assess its effect on symptoms, peak expiratory flow (PEF) and hydrogen peroxide (H2O2) in expired breath condensate as a marker of airway inflammation in patients with steroid-naive atopic asthma in a double-blind randomised, placebo-controlled clinical trial. Forty six patients with atopic asthma received two capsules of lipid extract (Lyprinol) or placebo b.i.d for 8 weeks. Each capsule of lipid extract contained 50 mg omega-3 polyunsaturated fatty acids and 100 mg olive oil, whereas placebo capsules contained only 150 mg olive oil. There was a significant decrease in daytime wheeze, the concentration of exhaled H2O2 and an increase in morning PEF in the lipid extract group compared to the placebo group. There were no significant side-effects. The authors conclude that lipid extract of New Zealand green-lipped mussel may have some beneficial effect in patients with atopic asthma.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12358334 L0903-1936 (Print) Clinical Trial Journal Article Randomized Controlled Trial12358334bHospital Therapeutic Clinic, Pavlov Medical University, St-Petersburg. Russia. emelav@netscape.net ~?XKharitonov, S. A. Donnelly, L. E. Montuschi, P. Corradi, M. Collins, J. V. Barnes, P. J.2002vDose-dependent onset and cessation of action of inhaled budesonide on exhaled nitric oxide and symptoms in mild asthma889-96Thorax5710Administration, Inhalation Adult Anti-Inflammatory Agents/*administration & dosage Asthma/*drug therapy/physiopathology Breath Tests Bronchodilator Agents/*administration & dosage Budesonide/*administration & dosage Carbon Monoxide/analysis Dose-Response Relationship, Drug Double-Blind Method Female Forced Expiratory Volume/physiology Humans Isoprostanes/analysis Male Nebulizers and Vaporizers Nitrates/analysis Nitric Oxide/*analysis Research Support, Non-U.S. Gov't S-Nitrosothiols/analysisOct+BACKGROUND: Dose dependent anti-inflammatory effects of inhaled corticosteroids in asthma are difficult to demonstrate in clinical practice. The anti-inflammatory effect of low dose inhaled budesonide on non-invasive exhaled markers of inflammation and oxidative stress were assessed in patients with mild asthma. METHODS: 28 patients entered a double blind, placebo controlled, parallel group study and were randomly given either 100 or 400 micro g budesonide or placebo once daily, inhaled from a dry powder inhaler (Turbohaler), for 3 weeks followed by 1 week without treatment. Exhaled nitric oxide (NO), exhaled carbon monoxide (CO), nitrite/nitrate, S-nitrosothiols, and 8-isoprostanes in exhaled breath condensate were measured four times during weeks 1 and 4, and once a week during weeks 2 and 3. RESULTS: A dose-dependent speed of onset and cessation of action of budesonide was seen on exhaled NO and asthma symptoms. Treatment with 400 micro g/day reduced exhaled NO faster (-2.06 (0.37) ppb/day) than 100 micro g/day (-0.51 (0.35) ppb/day; p<0.01). The mean difference between the effect of 100 and 400 micro g budesonide was -1.55 ppb/day (95% CI -2.50 to -0.60). Pretreatment NO levels were positively related to the subsequent speed of reduction during the first 3-5 days of treatment. Faster recovery of exhaled NO was seen after stopping treatment with budesonide 400 micro g/day (1.89 (1.43) ppb/day) than 100 micro g/day (0.49 (0.34) ppb/day, p<0.01). The mean difference between the effect of 100 and 400 micro g budesonide was 1.40 ppb/day (95% CI -0.49 to 2.31). Symptom improvement was dose-dependent, although symptoms returned faster in patients treated with 400 micro g/day. A significant reduction in exhaled nitrite/nitrate and S-nitrosothiols after budesonide treatment was not dose-dependent. There were no significant changes in exhaled CO or 8-isoprostanes in breath condensate. CONCLUSION: Measurement of exhaled NO levels can indicate a dose-dependent onset and cessation of anti-inflammatory action of inhaled corticosteroids in patients with mild asthma.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12324677 L0040-6376 (Print) Clinical Trial Journal Article Randomized Controlled Trial12324677Department of Thoracic Medicine, National Heart and Lung Institute, Imperial College School of Medicine, Royal Brompton & Harefield NHS Trust, London SW3 6LY, UK.~?Latzin, P. Griese, M.2002Exhaled hydrogen peroxide, nitrite and nitric oxide in healthy children: decrease of hydrogen peroxide by atmospheric nitric oxide353-8 Eur J Med Res78Adolescent Biological Markers Breath Tests Child Female Humans Hydrogen Peroxide/*metabolism Lung Diseases/diagnosis/metabolism Male Multivariate Analysis Nitric Oxide/*metabolism Nitrites/*metabolism Respiratory Function Tests Rhinitis, Allergic, Seasonal/*diagnosis/metabolismAug 30Hydrogen peroxide (H2O2) and nitrite (NO2-) in exhaled breath condensate have recently been suggested as non-invasive markers of airway inflammation. The goal of this study was to clarify the role of factors that may potentially influence the measurement of H2O2 and nitrite and to look for possible correlations among these inflammatory markers. H2O2 and nitrite values were assessed fluorometrically in breath condensate of 102 healthy children (age 4-18 years) and a detailed status of atopy (including history, lung function and skin prick test) was taken in all children. To find out the role of atmospheric nitric oxide, eNO and envNO were measured via chemiluminescence in association with the sampling of the breath condensate. Median (interquartile range) H2O2 was 0.51 (0.26 - 0.74) microM and nitrite was 3.3 (2.7 4.1) microM. A significant negative correlation between H2O2 and envNO was observed (r = -0.50; p < 0.0001). ENO was independent of envNO at our envNO range up to 56 ppb. No further correlation was found. The inflammatory markers in exhaled breath condensate H2O2, nitrite and eNO are not interrelated to each other in healthy children. Whereas eNO was not dependent on envNO values, high envNO values must be taken into account when measuring H2O2 in exhaled breath condensate.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12204843 !0949-2321 (Print) Journal Article12204843FChildren's Hospital, Ludwig Maximilians-University of Munich, Germany.|~?zCorradi, M. Alinovi, R. Goldoni, M. Vettori, M. Folesani, G. Mozzoni, P. Cavazzini, S. Bergamaschi, E. Rossi, L. Mutti, A.2002GBiomarkers of oxidative stress after controlled human exposure to ozone219-25 Toxicol Lett1341-3|Adult Biological Markers/blood Breath Tests Environmental Monitoring/*methods Exercise Test Female Genetic Predisposition to Disease Genotype Glutathione Transferase/genetics Humans Inhalation Exposure Lung/drug effects/physiology Male Oxidative Stress/*drug effects/physiology Ozone/*administration & dosage Quinone Reductases/genetics Research Support, Non-U.S. Gov't SpirometryAug 5This study was aimed at evaluating whether controlled short-term exposure to ozone (O(3)) induces changes in biomarkers of lung inflammation and oxidative stress in exhaled breath condensate (EBC) and blood of healthy subjects. Twenty-two volunteers were exposed to 0.1 ppm of O(3) for 2 h while performing moderate intermittent exercise. EBC and blood were collected before, immediately after and 18 h after exposure. Changes in biomarkers were measured both in EBC and blood, without significant alterations of lung function tests. Changes in EBC, but not in blood, were mainly accounted for by a subgroup of 'susceptible' individuals bearing the wild genotype for NAD(P)H:quinone oxidoreductase (NQO1) and the null genotype for glutathione-S-transferase M1 (GSTM1). Thus, a single 2-h exposure to 0.1 ppm of O(3) induces changes in biomarkers of inflammation and oxidative stress. Polymorphic NQO1 and GSTM1 act as modifier of the lung response to O(3).fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12191881 !0378-4274 (Print) Journal Article12191881Laboratory of Industrial Toxicology, Department of Clinical Medicine, Nephrology and Health Sciences, University of Parma, Via Gramsci 14, 43100 Parma, Italy. ~?AAntczak, A. Montuschi, P. Kharitonov, S. Gorski, P. Barnes, P. J.2002TIncreased exhaled cysteinyl-leukotrienes and 8-isoprostane in aspirin-induced asthma301-6Am J Respir Crit Care Med1663Adult Anti-Inflammatory Agents, Non-Steroidal/*adverse effects Aspirin/*adverse effects Asthma/*chemically induced/*metabolism *Breath Tests Cysteine/*analysis *Dinoprost/*analogs & derivatives Dinoprostone/analysis F2-Isoprostanes/*analysis Female Humans Inflammation Mediators/*analysis Leukotriene B4/analysis Leukotrienes/*analysis Male Middle Aged Oxidative Stress/physiology Oxytocics/analysis Research Support, Non-U.S. Gov't Respiratory System/drug effects/*metabolism Vasoconstrictor Agents/*analysisAug 1The pathogenesis of aspirin-induced asthma (AIA) has not yet been clearly elucidated, although eicosanoid metabolites appear to play an important role. We hypothesized that levels of eicosanoids in exhaled air condensate are abnormal in patients with AIA and that they change in patients receiving steroid therapy. We measured cysteinyl-leukotrienes (cys-LTs), prostaglandin E(2) (PGE(2)), and leukotriene B(4) (LTB(4)), and also 8-isoprostane as a marker of oxidative stress, by enzyme immunoassay in exhaled breath condensate from patients with AIA (17 steroid naive; mean age, 41 +/- 23 years; FEV(1), 63%pred), 26 patients with aspirin-tolerant asthma (ATA) (11 steroid naive; mean age, 47 +/- 18 years; FEV(1), 69%pred), and 16 healthy subjects (mean age, 45 +/- 17 years; FEV(1), 93%pred). Cys-LTs were significantly higher in steroid-naive patients with AIA compared with steroid-naive patients with ATA and healthy subjects (152.3 +/- 30.4 and 36.6 +/- 7.1 versus 19.4 +/- 2.8 pg/ml; p < 0.05 and p < 0.05, respectively). Steroid-naive patients with AIA also had higher levels of 8-isoprostane than normal subjects (131.8 +/- 31.0 versus 21.9 +/- 4.5 pg/ml; p < 0.05). There were significantly lower levels of both cys-LTs and 8-isoprostanes in steroid-treated patients with AIA. There was no difference in either the PGE(2) or LTB(4) level between the patient groups. This is the first study to show that cys-LTs and 8-isoprostanes are elevated in expired breath condensate of steroid-naive patients with AIA, and that cys-LTs are decreased in steroid-treated patients. Exhaled PGE(2) levels are not reduced, so that it is unlikely that a deficiency of PGE(2) is an important mechanism, whereas exhaled LTB(4) levels are unchanged, indicating an abnormality beyond 5-lipoxygenase.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12153961 !1073-449X (Print) Journal Article12153961sDepartment of Thoracic Medicine, National Heart and Lung Institute, Imperial College, London, UK. magant@kki.net.pl~? Montuschi, P.2002(Indirect monitoring of lung inflammation238-42Nat Rev Drug Discov13Biological Markers/analysis Humans Hydrogen Peroxide/analysis Immunoenzyme Techniques Inflammation/*diagnosis/physiopathology Lung Diseases/*diagnosis/physiopathology Monitoring, Physiologic/methods Reproducibility of Results Research Support, Non-U.S. Gov'tMarThe assessment of airway inflammation by non-invasive methods could provide a signal to start anti-inflammatory treatment before the onset of symptoms and the impairment of lung function. It could also be useful in the follow-up of patients with lung disease, and for guiding drug treatment. Measuring inflammatory markers in exhaled breath condensate is potentially the easiest way to quantify lung inflammation. The clinical applicability of this method could facilitate the practice of respiratory medicine.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12120509 (1474-1776 (Print) Journal Article Review12120509Department of Pharmacology, School of Medicine, Catholic University of the Sacred Heart, L.go F. Vito 1-00168, Rome, Italy. p.montuschi@ic.ac.uk~?WSchreiber, J. Meyer, C. Rusch-Gerdes, S. Richter, E. Beck, H. Fischer, J. F. Rosahl, W.2002eMycobacterium tuberculosis gene-amplification in breath condensate of patients with lung tuberculosis290-1 Eur J Med Res76Adult Aged *Breath Tests Female Gene Amplification Humans Ligase Chain Reaction Male Middle Aged Mycobacterium tuberculosis/*genetics/*isolation & purification Tuberculosis, Pulmonary/*diagnosisJun 28The applicability of Mycobacterium tuberculosis-gene-amplification test (NAT) in breath condensate (BC) was examined in lung tuberculosis (TB). Ten patients with bacteriologically confirmed lung TB have been examined by ligase chain reaction (LCx). In BC the NAT were negative in all patients as well as the examination of acid fast smears and of solid phase or liquid media cultures were. This indicates that the use of NAT in BC can not replace or complement sputum or materials obtained invasively in patients with lung TB.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12117666 !0949-2321 (Print) Journal Article12117666mMunicipal Hospital Dessau, Dept. of Pulmonology, Auenweg 38, D-06822 Dessau, Germany. skd.innere2@t-online.de ~?RCarpagnano, G. E. Resta, O. Foschino-Barbaro, M. P. Gramiccioni, E. Carpagnano, F.2002[Interleukin-6 is increased in breath condensate of patients with non-small cell lung cancer141-5Int J Biol Markers172Aged *Breath Tests Carcinoma, Non-Small-Cell Lung/diagnosis/*metabolism Female Humans Interleukin-6/analysis/*metabolism Lung Neoplasms/diagnosis/*metabolism Male Middle AgedApr-JunDespite recent advances in the diagnosis and treatment of non-small cell lung cancer (NSCLC), most patients still present with advanced stage disease at the time of diagnosis. Recent studies suggest that IL-6 is involved in the development of lung cancer. The aim of the present study was to investigate whether the measurement of IL-6 levels in the breath condensate of NSCLC patients could be used to bring forward the moment of diagnosis and to monitor the progression of the disease. Twenty patients with histological evidence of NSCLC (14 men and 6 women, age 63+/-8 years) and 15 healthy controls (8 men and 7 women, age 45+/-6 years) were enrolled in the study. IL6 was measured in the exhaled breath condensate of patients and controls by means of a specific enzyme immunoassay kit. Higher concentrations of exhaled IL-6 were found in NSCLC patients (9.6+/-0.3 pg/mL) than in controls (3.5+/-0.2 pg/mL). A statistically significant difference was observed between patients with NSCLC at different stages: higher concentrations of IL-6 (10.9+/-0.5 pg/mL) were found in patients with metastatic disease than in those with stage III (9.7+/-0.4 pg/mL), stage II (8.9+/-0.3 pg/mL) and stage I disease (7.9+/-0.3 pg/mL). These findings suggest that the measurement of IL-6 in the breath condensate of patients with NSCLC could be proposed as a parameter to take into account in early diagnosis and disease monitoring.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12113582 !0393-6155 (Print) Journal Article12113582=Institute of Respiratory Diseases, University of Bari, Italy. +~?"Griese, M. Noss, J. von Bredow, C.20027Protein pattern of exhaled breath condensate and saliva690-6 Proteomics26Adult Breath Tests/*methods Electrophoresis, Gel, Two-Dimensional/*methods Humans Middle Aged Respiration Saliva/*metabolism Salivary Proteins/*analysis/*chemistry SoftwareJunThe proteins recovered in exhaled breath condensate (EBC) might be used to non-invasively monitor respiratory diseases. However, the range of proteins and their source are still unresolved and contamination by saliva or a similar protein pattern in the nasal and bronchial compartments may make interpretation of the data difficult. We studied nasal EBC (collected through a "free of touch" technique by negative pressure), oral tidal, and oral forced EBC (collected through a rebreathing valve as a saliva trap connected to tubing submerged into ice) and matched saliva samples from five healthy adult subjects. The protein samples were separated by two-dimensional electrophoresis and the silver stained gels were analyzed by Melanie 2 software. In both nasal and oral EBC, three spots (72 kDa/isoelectric point (pI) 6.6-7.0, 66 kDa/pI 5.9-6.7 and 45-48 kDa/pI 8.0-8.6) were consistently present in all subjects. Several other proteins were only sporadically detected. Despite improbable saliva contamination (no phosphorus contamination in the same oral and nasal EBC, no amylase activity in 10 pairs of nasal and oral EBC collected by the same technique), on average 63% and 71% of the spots identified in oral and nasal EBC were also found in the matched individual saliva samples. Compared to saliva, the range and amount of protein in all types of EBC was very small. Even when collected free of saliva contamination the majority of proteins present in EBC was also found in saliva, suggesting that these proteins are present in both compartments, e.g. saliva and secretions of the lower airspaces. The quantification and identification of specific proteins in the various compartments is warranted in future studies to determine the practical value of EBC.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12112850 !1615-9853 (Print) Journal Article12112850fChildrens' Hospital, Ludwig Maximilians-University of Munich, Germany. griese@pk-i.med.uni-muenchen.de@~?Hunt, J.2002VExhaled breath condensate: an evolving tool for noninvasive evaluation of lung disease28-34J Allergy Clin Immunol1101rBiological Markers/analysis Breath Tests/instrumentation/*methods Child, Preschool Humans Lung Diseases/*diagnosisJulExhaled breath condensate (EBC) contains aerosolized airway lining fluid and volatile compounds that provide noninvasive indications of ongoing biochemical and inflammatory activities in the lung. Rapid increase in interest in EBC has resulted from the recognition that in lung disease this easily sampled fluid has measurable characteristics that differ prominently from health. These assays have provided evidence of airway and lung redox deviation, acid-base status, and degree and type of inflammation in acute and chronic asthma, chronic obstructive pulmonary disease, adult respiratory distress syndrome, occupational diseases, and cystic fibrosis. Characterized by uncertain and variable degrees of dilution, EBC does not provide precise assessment of individual solute concentrations within native airway lining fluid. However, it can provide useful information when concentrations differ substantially between health and disease or are based on ratios of solutes found in the sample. Because they can be used to measure the targets of modern therapy, EBC assays are likely to become integral components of future clinical studies, and after further technical work is accomplished, they might be used to diagnose and monitor therapy in individual patients.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12110814 (0091-6749 (Print) Journal Article Review12110814Division of Pediatric Respiratory Medicine and the Asthma and Allergic Diseases Center, University of Virginia, Charlottesville, VA 22908, USA. |~?Kharitonov, S. A. Barnes, P. J.20027Biomarkers of some pulmonary diseases in exhaled breath1-32 Biomarkers71Ammonia/analysis Asthma/diagnosis Biological Markers Breath Tests/*methods Carbon Monoxide/analysis Cystic Fibrosis/diagnosis Humans Hydrogen Peroxide/analysis Lipid Peroxidation Lung Diseases/*diagnosis Nitric Oxide/analysis Pulmonary Disease, Chronic Obstructive/diagnosisJan-FebAnalysis of various biomarkers in exhaled breath allows completely non-invasive monitoring of inflammation and oxidative stress in the respiratory tract in inflammatory lung diseases, including asthma, chronic obstructive pulmonary disease (COPD), cystic fibrosis (CF), bronchiectasis and interstitial lung diseases. The technique is simple to perform, may be repeated frequently, and can be applied to children, including neonates, and patients with severe disease in whom more invasive procedures are not possible. Several volatile chemicals can be measured in the breath (nitric oxide, carbon monoxide, ammonia), and many non-volatile molecules (mediators, oxidation and nitration products, proteins) may be measured in exhaled breath condensate. Exhaled breath analysis may be used to quantify inflammation and oxidative stress in the respiratory tract, in differential diagnosis of airway disease and in the monitoring of therapy. Most progress has been made with exhaled nitric oxide (NO), which is increased in atopic asthma, is correlated with other inflammatory indices and is reduced by treatment with corticosteroids and antileukotrienes, but not (beta 2-agonists. In contrast, exhaled NO is normal in COPD, reduced in CF and diagnostically low in primary ciliary dyskinesia. Exhaled carbon monoxide (CO) is increased in asthma, COPD and CF. Increased concentrations of 8-isoprostane, hydrogen peroxide, nitrite and 3-nitrotyrosine are found in exhaled breath condensate in inflammatory lung diseases. Furthermore, increased levels of lipid mediators are found in these diseases, with a differential pattern depending on the nature of the disease process. In the future it is likely that smaller and more sensitive analyzers will extend the discriminatory value of exhaled breath analysis and that these techniques may be available to diagnose and monitor respiratory diseases in the general practice and home setting.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12101782 (1354-750X (Print) Journal Article Review12101782Department of Thoracic Medicine, National Heart and Lung Institute, Faculty of Medicine, Imperial College, Royal Brompton Hospital, London, UK. s.kharitonov@ic.ac.uk ~??van Beurden, W. J. Dekhuijzen, P. N. Harff, G. A. Smeenk, F. W.2002]Variability of exhaled hydrogen peroxide in stable COPD patients and matched healthy controls211-6 Respiration693Aged Aged, 80 and over Breath Tests Circadian Rhythm/*physiology Cotinine/analysis Female Humans Hydrogen Peroxide/*analysis Male Middle Aged Oxidative Stress/physiology Pulmonary Disease, Chronic Obstructive/*diagnosis/metabolism/*physiopathology@BACKGROUND: Because inflammation induces oxidative stress, exhaled hydrogen peroxide (H(2)O(2)), which is a marker of oxidative stress, may be used as a non-invasive marker of airway inflammation in chronic obstructive pulmonary disease (COPD). There are no data on the circadian variability of exhaled H(2)O(2) in COPD patients. OBJECTIVE: The aim of this study was to investigate the variability of the H(2)O(2) concentration in breath condensate of stable COPD patients and of matched healthy control subjects. METHODS: We included 20 patients with stable mild COPD (forced expiratory volume in 1 s approximately 70% of predicted) and 20 healthy subjects, matched for age, sex and pack-years, all smokers or ex-smokers. Breath condensate was collected and its H(2)O(2) concentration determined fluorometrically three times on day 0 (9 and 12 a.m., and 3 p.m.) and once on days 1, 2, 3, 8 and 21. RESULTS: The mean H(2)O(2) concentration increased significantly during the day in both the patient and control groups (p = 0.02 and p < 0.01, respectively). Over a longer period up to 21 days, the mean concentration did not change in both groups. There was no significant difference between patients and controls. The mean coefficient of variation over 21 days was 45% in the patient group and 43% in the control group (p = 0.8). CONCLUSIONS: The exhaled H(2)O(2) concentration increased significantly during the day in both stable COPD patients and controls. Over a period of 3 weeks, the mean H(2)O(2) concentration did not change and the variability within the subjects was similar in both groups.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12097763 !0025-7931 (Print) Journal Article12097763mDepartment of Pulmonology, Catharina Hospital Eindhoven, The Netherlands. w.beurden@researchlab-long.demon.nl 5~?OKostikas, K. Papatheodorou, G. Ganas, K. Psathakis, K. Panagou, P. Loukides, S.2002MpH in expired breath condensate of patients with inflammatory airway diseases1364-70Am J Respir Crit Care Med16510Adult Aged Analysis of Variance Asthma/*diagnosis/physiopathology Breath Tests Bronchiectasis/*diagnosis/physiopathology Case-Control Studies Comparative Study Confidence Intervals Female Humans *Hydrogen-Ion Concentration Male Middle Aged Predictive Value of Tests Probability Prognosis Prospective Studies Pulmonary Disease, Chronic Obstructive/*diagnosis/physiopathology Reference Values Respiratory Function Tests Sensitivity and Specificity Severity of Illness Index Sputum/chemistry Statistics, NonparametricMay 15@Endogenous airway acidification, as assessed by pH in expired breath condensate, has been implicated in asthma pathophysiology. We measured pH in breath condensate of patients with inflammatory airway diseases in stable condition and examined its relationship with the inflammatory process (as assessed by differential cell counts in induced sputum), oxidative stress (as assessed by H(2)O(2) and 8-isoprostane), and nitric oxide metabolism (as assessed by total nitrate/nitrite). We studied 40 patients with bronchial asthma (20 with moderate disease, forced expiratory volume in 1 second 60 [10]% SD predicted), 20 patients with bronchiectasis, 20 patients with chronic obstructive pulmonary disease (COPD), and 10 normal subjects. Mean (95% confidence intervals) pH values were significantly lower in patients with COPD and bronchiectasis compared with patients with asthma and control subjects (7.16, 7.09-7.23 and 7.11, 7.04-7.19 versus 7.43, 7.35-7.52 and 7.57, 7.51-7.64, respectively, p < 0.0001). Patients with moderate asthma had significantly lower values compared with mild and control subjects. In patients with COPD and bronchiectasis, the values of pH were significantly correlated with both sputum neutrophilia and oxidative stress. Respectively, in patients with moderate asthma, a significant correlation was observed between pH and sputum eosinophilia, total nitrate/nitrite, and oxidative stress. The pH of the expired breath condensate might be a simple, noninvasive, inexpensive, and easily repeatable procedure for the evaluation of the inflammatory process in airway diseases.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12016097 !1073-449X (Print) Journal Article12016097dDepartment of Pneumonology and Clinical Research Unit, Athens Army General Hospital, Athens, Greece.~?wKirsanov, A. I. Dolgodvorov, A. F. Leont'ev, V. G. Gorbacheva, I. A. Romanova, V. D. Velichko, L. S. Aleksandrov, V. V.2001;[Chemical element levels in various human biological media]16-20Klin Lab Diagn3 Asthma/metabolism Blood Chemical Analysis Body Fluids/*chemistry Breath Tests Electrolytes/analysis English Abstract Erythrocytes/chemistry Female Humans Male Metals/analysis Phosphorus/analysis Saliva/chemistry Sulfhydryl Compounds/analysis Sulfur/analysis UrinalysisMarAnalysis of element composition (sodium, potassium, magnesium, calcium, zinc, copper, iron, total sulfur and its compounds, chloride, and inorganic phosphorus) in 40 normal subjects (22 women and 18 men aged 17-30 years) showed that the composition is specific for each of the analyzed media (blood, plasma, erythrocytes, urine, saliva, exhaled air condensate). Element composition of exhaled air condensate is described for the first time. Analysis of element composition in 102 patients with asthma demonstrated characteristic shifts in the concentrations of some macro- and trace elements, which confirms the clinical significance of analysis and correction of deviations in the element composition of biological media.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12014071 !0869-2084 (Print) Journal Article12014071TKontsentratsiia khimicheskikh elementov v raznykh biologicheskikh sredakh cheloveka.~?Montuschi, P. Barnes, P. J.2002HAnalysis of exhaled breath condensate for monitoring airway inflammation232-7Trends Pharmacol Sci235Breath Tests/*methods Humans Hydrogen Peroxide/analysis Inflammation/*diagnosis Inflammation Mediators/*analysis Isoprostanes/analysis Lung Diseases/*diagnosis Nitric Oxide/analysis Oxidative Stress Prostaglandins/analysis Research Support, Non-U.S. Gov'tMay;Several inflammatory mediators have been identified in the exhaled breath condensate (EBC) that is formed by breathing through a cooling system. Analysis of EBC is a noninvasive method that allows repeat measurements of lung inflammation and is potentially useful for monitoring drug therapy. Characterization of the profiles of exhaled markers could help to discriminate between different inflammatory lung diseases; thus, EBC might be a novel, noninvasive approach to monitoring lung diseases. However, several methodological issues, such as standardization of sample collection and validation of analytical techniques, need to be addressed before this method can be applied clinically. Controlled studies are needed to establish the utility of EBC markers for guiding pharmacological treatment in inflammatory lung diseases.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=12008001 (0165-6147 (Print) Journal Article Review12008001zDept of Pharmacology, School of Medicine, Catholic University of the Sacred Heart, 00168 Rome, Italy. p.montuschi@ic.ac.uk~?DShahid, S. K. Kharitonov, S. A. Wilson, N. M. Bush, A. Barnes, P. J.2002kIncreased interleukin-4 and decreased interferon-gamma in exhaled breath condensate of children with asthma1290-3Am J Respir Crit Care Med1659Adolescent Asthma/*diagnosis/immunology Biological Markers *Breath Tests Case-Control Studies Child Child, Preschool Female Humans Interferon Type II/*analysis/metabolism Interleukin-4/*analysis/metabolism Male Research Support, Non-U.S. Gov'tMay 1Exhaled breath condensate analysis for noninvasive quantification of airway inflammation in asthma is a potentially useful research tool in children. There is an imbalance between T-helper (Th)-2 cells, which secrete interleukin (IL)-4, and Th1 cells, which secrete interferon (IFN)-gamma, in asthma. We measured concentrations of IL-4 and IFN-gamma in breath condensates of 37 children (11 normal, 12 steroid-naive, and 14 steroid-treated children with asthma). Exhaled IFN-gamma was significantly lower in steroid-naive and steroid-treated children with asthma compared with normal control subjects (3.7 +/- 0.2 versus 5.1 +/- 0.4 pg/ml, p < 0.01 and 4.1 versus 5.1 pg/ml, p < 0.05). By contrast, mean exhaled IL-4 was elevated in asthma (53.7 +/- 4.2 pg/ml) compared with normal children (35.7 +/- 6.2 pg/ml, p < 0.05) and concentrations were lower with steroid treatment (37.5 +/- 5.6 pg/ml, p < 0.05). Exhaled IL-4 was significantly lower in children with asthma on more than 600 microg inhaled steroid/day. The IL-4/IFN-gamma ratio was significantly greater in children with asthma compared with control children and the children with asthma on inhaled steroid therapy. We have shown for the first time that IFN-gamma and IL-4 can be assayed in exhaled breath condensate and shows an increased ratio of IL-4/IFN-gamma, consistent with predominance of Th2 cells in airways of children with asthma. Exhaled breath condensate analysis may have a useful role in studying allergic inflammation in childhood asthma.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11991881 !1073-449X (Print) Journal Article11991881tDepartment of Thoracic Medicine, National Heart and Lung Institute, Royal Brompton Hospital, London, United Kingdom._~?Kharitonov, S. A. Barnes, P. J.2001Exhaled markers of inflammation217-24Curr Opin Allergy Clin Immunol13Biological Markers/analysis *Breath Tests Carbon Monoxide/analysis Humans Hydrocarbons/analysis Inflammation/*diagnosis/metabolism Leukotrienes/analysis Lung Diseases/diagnosis Nitric Oxide/analysis Oxidative Stress Prostaglandins/analysis TemperatureJunExhaled markers of inflammation allow completely noninvasive monitoring of inflammation and oxidative stress in the respiratory tract in inflammatory lung diseases, including asthma, chronic obstructive pulmonary disease, cystic fibrosis, bronchiectasis and interstitial lung diseases. Such noninvasive techniques are simple to perform, may be repeated frequently and can be applied in children, including neonates and patients with severe disease in whom more invasive procedures are not possible. Several volatile chemicals can be measured in the breath (nitric oxide, carbon monoxide, hydrocarbons), and many nonvolatile molecules (mediators, oxidation and nitration products, proteins) may be measured in exhaled breath condensate.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11964692 !1528-4050 (Print) Journal Article11964692VNational Heart and Lung Institute, Imperial College, London, UK. s.kharitonov@ic.ac.uk !~?Montuschi, P. Barnes, P. J.20021Exhaled leukotrienes and prostaglandins in asthma615-20J Allergy Clin Immunol10942Adult Asthma/diagnosis/*metabolism/physiopathology *Breath Tests Cross-Sectional Studies Female Humans Leukotriene B4/analysis/*metabolism Leukotriene E4/analysis/*metabolism Lung/physiopathology Male Nitric Oxide/analysis/metabolism Prostaglandins/analysis/*metabolism Research Support, U.S. Gov't, P.H.S.AprSBACKGROUND: Most of the studies investigating the role of leukotrienes (LTs) and prostaglandins (PGs) in asthma have used invasive (eg, bronchoalveolar lavage fluid) or semi-invasive (eg, sputum induction) techniques. Others have measured eicosanoids in plasma or urine, probably reflecting systemic rather than lung inflammation. Collection of exhaled breath condensate (EBC) is a noninvasive method to collect airway secretions. OBJECTIVE: We sought to investigate whether eicosanoids are measurable in EBC, to show possible differences in their concentrations in asthmatic patients and healthy subjects, and to investigate whether exhaled eicosanoids correlate with exhaled nitric oxide (NO), a marker of airway inflammation. METHODS: Twelve healthy nonsmokers and 15 steroid-naive patients with mild asthma were studied. Subjects attended on one occasion for pulmonary function tests, collection of EBC, and exhaled NO measurements. Exhaled LTB(4)-like immunoreactivity, LTE(4)-like immunoreactivity, PGE(2)-like immunoreactivity, PGD(2)-methoxime, PGF(2)(alpha)-like immunoreactivity, and thromboxane B(2)-like immunoreactivity were measured by means of enzyme immunoassays. RESULTS: LTE(4)-like immunoreactivity and LTB(4)-like immunoreactivity were detectable in EBC in healthy subjects, and their levels in asthmatic patients were increased about 3-fold (P <.0001) and 2-fold (P <.0005), respectively. Exhaled NO was increased in asthmatic patients compared with healthy subjects (P <.0001). There was a correlation between exhaled LTB(4) and exhaled NO (r = 0.56, P <.04) in patients with asthma. When measurable, prostanoid levels were similar in asthmatic patients and control subjects. CONCLUSIONS: Exhaled LTE(4) and LTB(4) are increased in steroid-naive patients with mild asthma. EBC may be proved to be a novel method to monitor airway inflammation in asthma.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11941309 !0091-6749 (Print) Journal Article11941309eDepartment of Pharmacology, School of Medicine, Catholic University of the Sacred Heart, Rome, Italy.B~?JFormanek, W. Inci, D. Lauener, R. P. Wildhaber, J. H. Frey, U. Hall, G. L.2002KElevated nitrite in breath condensates of children with respiratory disease487-91 Eur Respir J193Adolescent Analysis of Variance Asthma/*metabolism/physiopathology Biological Markers/analysis Breath Tests Case-Control Studies Child Child, Preschool Comparative Study Cough/*metabolism/physiopathology Cystic Fibrosis/*metabolism/physiopathology Female Humans Male Nitric Oxide/*analysis/metabolism Nitrites/*analysis Prognosis Prospective Studies Reference Values Research Support, Non-U.S. Gov't Respiratory Function Tests Sensitivity and SpecificityMarThe aim of the study was to determine the differences in nitrite, in the exhaled breath condensates of healthy children and those children with asthma, cystic fibrosis (CF) and nonasthmatic, episodic cough. Breath condensates were obtained from 66 children (43 males:23 females, 3.1-16 yrs) and included 29 asthmatics, 12 clinically stable CF patients, 12 children with cough but not asthma and 13 healthy volunteers. The collected condensate was assayed colourimetrically using the Griess reaction to determine nitrite concentrations. Patients with CF (median: 5-95% percentiles; 2.02: 0.43-6.37 microM) or asthma (2.10: 0.63-5.45 microM) had significantly higher levels of nitrite compared to healthy subjects (0.41: 0.13-1.83 microM; p<0.05) or subjects with cough (0.75: 0.03-1.75 microM; p<0.05). Airway inflammation, as assessed by the nitrite in breath condensates, is present in children with asthma and cystic fibrosis, but not those children with nonasthmatic, episodic cough. Nitrite can be conveniently, cheaply and rapidly measured in breath condensates of children as young as 3 yrs of age, and may prove useful for the assessment of airway inflammation in children with respiratory disease.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11936527 !0903-1936 (Print) Journal Article11936527RDept of Respiratory Medicine, University Children's Hospital, Zurich, Switzerland.~?dvan Beurden, W. J. Harff, G. A. Dekhuijzen, P. N. van den Bosch, M. J. Creemers, J. P. Smeenk, F. W.2002aAn efficient and reproducible method for measuring hydrogen peroxide in exhaled breath condensate197-203 Respir Med963Aged Breath Tests/instrumentation/*methods Cryopreservation Female Humans Hydrogen Peroxide/*analysis Male Middle Aged Pulmonary Disease, Chronic Obstructive/*diagnosis Reproducibility of Results Research Support, Non-U.S. Gov't Sensitivity and SpecificityMarWe investigated the sensitivity and reproducibility of a test procedure for measuring hydrogen peroxide (H202) in exhaled breath condensate and the effect of storage of the condensate on the H2O2 concentration, and compared the results to previous studies.Twenty stable COPD patients breathed into our collecting device twice for a period of 10 min.The total exhaled air volume (EAV) and condensate volume were measured both times and the H2O2 concentration of the condensate was determined fluorimetrically.The concentration was measured again after freezing the reaction product at -70 degrees C for a period of 10, 20 and 40 days. We collected 2-5 ml condensate in 10 min. The EAV and condensate volumes were strongly correlated. There was no significant difference between the mean H2O2 concentration of the first and second test. We obtained a detect on limit for the H2O2 concentration of 0.02 micromoll(-1). The H2O2 concentration appeared to remain stable for a period up to 40 days of freezing. Compared to previous studies we developed a more efficient breath condensate collecting device and obtained a lower H2O2 detection limit.The measurement of exhaled H2O2 was reproducible. In addition, storage of the samples up to 40 days showed no changes in H2O2 concentration.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11908513 !0954-6111 (Print) Journal Article11908513nDepartment of Pulmonology, Catharina Hospital, Eindhoven, The Netherlands. w.beurden@researchlab-long.demon.nl +~?dEffros, R. M. Hoagland, K. W. Bosbous, M. Castillo, D. Foss, B. Dunning, M. Gare, M. Lin, W. Sun, F.20026Dilution of respiratory solutes in exhaled condensates663-9Am J Respir Crit Care Med1655Acid-Base Equilibrium/*physiology Adult Breath Tests/*methods Female Humans Inflammation Mediators/analysis Lung Diseases/*diagnosis/physiopathology Male Middle Aged Pulmonary Disease, Chronic Obstructive/diagnosis/physiopathology Reference Values Research Support, U.S. Gov't, P.H.S. Respiratory Mucosa/*physiopathology Sensitivity and Specificity Sleep Apnea, Obstructive/diagnosis/physiopathology Tracheostomy Water-Electrolyte Balance/*physiologyMar 1hMost exhaled water is produced as gaseous water vapor, which can be collected in cooled condensers. The presence of nonvolatile solutes in these condensates suggests that droplets of respiratory fluid (RF) have also been collected. However, calculation of RF solute concentrations from condensates requires estimation of the dilution of RF droplets by water vapor. We used condensate electrolyte concentrations to calculate the dilution of RF droplets in condensates from 20 normal subjects. The total ionic concentration (conductivity) was 497 plus minus 68 (mean plus minus SEM) muM. Of this, 229 plus minus 43 muM was NH(4)(+), but little NH(4)(+) was collected from subjects with tracheostomies, indicating oral formation. The Na+ concentration in condensate ([Na+](cond)) averaged 242 plus minus 43 muM. Large variations in [Na(+)](cond) correlated well with variations of K+ in condensate ([K+](cond)) and Cl-) in condensate ([Cl-](cond)), and were attributed to differences in respiratory droplet dilution. Dividing condensate values of ([Na+] + [K+] ) by those of plasma indicated that RF represented between 0.01% and 2.00% of condensate volumes. Calculated values for Na+, K+, Cl-, lactate, and protein in RF were [Na+](RF) = 91 +/- 8 mM, [K+](RF) = 60 +/- 11 mM, [Cl-](RF) = 102 +/- 17 mM, [lactate](RF) = 44 +/- 17 mM, and [protein](RF) = 7.63 +/- 1.82 g/dl, respectively.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11874811 !1073-449X (Print) Journal Article11874811xDepartment of Medicine, Medical College of Wisconsin, 9200 West Wisconsin Ave., Milwaukee, WI 53226, USA. effros@mcw.edu~?INayeri, F. Millinger, E. Nilsson, I. Zetterstrom, Brudin, L. Forsberg, P.2002SExhaled breath condensate and serum levels of hepatocyte growth factor in pneumonia115-9 Respir Med962/Adult Aged Aged, 80 and over Analysis of Variance Biological Markers/analysis/blood Breath Tests Bronchi/chemistry Case-Control Studies Female Hepatocyte Growth Factor/*analysis/blood Humans Male Middle Aged Pneumonia/*metabolism Pulmonary Alveoli/chemistry Research Support, Non-U.S. Gov't Time FactorsFebHepatocyte growth factor (HGF) is a protein produced by mesenchymal cells in many organs, which can stimulate epithelial growth. An enhanced production and concentration of HGF is observed after injuries. The lung is one of the major sources of HGF. By cooling exhaled air, a condensate is formed containing molecules from bronchi and alveoli. In order to investigate HGF-concentration and time course in pneumonia, paired serum and exhaled breath condensate was collected from 10 patients with pneumonia, 10 patients with non-respiratory infections and 11 healthy controls. The concentration of HGF was measured by an immunoassay kit. In the acute phase HGF-levels in breath condensate and serum were significantly higher in the patients with pneumonia compared to the control groups. Similar concentrations in breath condensate were seen in healthy controls and in patients with non-respiratory infections. In the patients with pneumonia a decrease in serum HGF was seen already after 4-7 days while HGF values in breath condensate remained elevated even after 4-6 weeks. These results might imply local product on of HGF in the lungs and a long repair and healing process after pneumonia.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11860168 !0954-6111 (Print) Journal Article11860168bPulmonary Department, Allergy Center, University Hospital, Linkoping, Sweden. fariba.nayeri@lio.se ~?ELoukides, S. Bouros, D. Papatheodorou, G. Panagou, P. Siafakas, N. M.2002pThe relationships among hydrogen peroxide in expired breath condensate, airway inflammation, and asthma severity338-46Chest1212Asthma/pathology/*physiopathology Blood Proteins/analysis *Breath Tests Eosinophil Granule Proteins Eosinophils/metabolism/pathology Humans Hydrogen Peroxide/*analysis Inflammation Neutrophils/metabolism/pathology *Ribonucleases Severity of Illness Index Sputum/cytologyFeb STUDY OBJECTIVE: To investigate which cells are the main source of hydrogen peroxide (H(2)O(2)) production in stable patients with asthma and the associations among H(2)O(2) levels, airway inflammation, and disease severity. SETTING: Inpatient respiratory unit and outpatient clinic in tertiary-care hospital. PATIENTS: Fifty stable asthmatic patients with disease severity ranging from mild to moderate. METHODS: H(2)O(2) was measured in expired breath condensate and was correlated with variables expressing both asthma severity (ie, FEV(1) percent predicted, peak expiratory flow rate [PEFR] variability, symptom score, and histamine airways responsiveness) and airway inflammation (ie, differential cell counts from induced sputum and levels of eosinophil cationic protein [ECP]). RESULTS: The mean (95% confidence interval [CI]) concentration of H(2)O(2) was significantly elevated in patients with asthma compared to that in control subjects (mean, 0.67 microM [95% CI, 0.56 to 0.77 microM] vs 0.2 microM [95% CI, 0.16 to 0.24 microM]; p < 0.0001). The difference was primarily due to the elevation of H(2)O(2) in patients with moderate asthma whose expired breath H(2)O(2) level of 0.95 microM (95% CI, 0.76 to 1.12 microM) was significantly higher from that of patients with mild-persistent and mild-intermittent asthma (mean, 0.59 microM [95% CI, 0.47 to 0.7 microM] and 0.27 [95% CI, 0.23 to 0.32 microM], respectively; p < 0.0001). H(2)O(2) concentration was positively related to sputum eosinophilia as well as to ECP concentration. A similar correlation was found between H(2)O(2) and neutrophils in patients with moderate asthma. A positive correlation was observed between H(2)O(2) level, symptom score, and PEFR variability. H(2)O(2) level was negatively related to FEV(1) percent predicted. Further analysis showed that only patients with moderate asthma who were not receiving inhaled steroids were found to have a strong relationship with the variables tested. CONCLUSIONS: Eosinophils are the predominate cells that generate H(2)O(2) in all forms of the disease, while neutrophils might be responsible for the highest levels that are observed in the more severe forms of the disease. The role of H(2)O(2) concentration in predicting the severity of the disease as well as in the inflammatory process is limited and depends on the use of inhaled steroid therapy and the classification of the severity of the disease.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11834641 !0012-3692 (Print) Journal Article11834641pDepartment of Pneumonology and Clinical Research Unit, Athens Army General Hospital, Athens, Greece. ssat@hol.gr~?0Larstad, M. Ljungkvist, G. Olin, A. C. Toren, K.2002{Determination of malondialdehyde in breath condensate by high-performance liquid chromatography with fluorescence detection107-14-J Chromatogr B Analyt Technol Biomed Life Sci7661Asthma/*metabolism *Breath Tests Case-Control Studies Chromatography, High Pressure Liquid/*methods Female Humans Male Malondialdehyde/*metabolism Reproducibility of Results Research Support, Non-U.S. Gov't Sensitivity and Specificity Spectrometry, FluorescenceJan 5An automated and rapid method for quantifying malondialdehyde (MDA) in breath condensate was developed and validated. The method is based on derivatisation with thiobarbituric acid, HPLC separation and fluorescence detection and is optimised for determination of MDA in breath condensate. Sample collection is non-invasive and simple. The detection limit (4.1 nM) is low, precision is good and the analysis time is short. The response is linear in the concentration range of 0.020 to 1.0 microM. Samples could be stored for 1 month at -20 degrees C and for 3 months at -80 degrees C without losses. Using this method, there was no statistically significant difference between patients with asthma and patients without asthma. However, among females, subjects with asthma had higher MDA levels as compared to females without asthma (0.17 vs. 0.12 pmol/s, p=0.04). The use of the method when studying airway inflammation has to be further evaluated.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11820285 !1570-0232 (Print) Journal Article11820285Department of Occupational Medicine, The Sahlgrenska Academy at Goteborg University, Gothenburg, Sweden. monica.larstad@ymk.gu.seZ~?YJobsis, R. Q. Schellekens, S. L. Fakkel-Kroesbergen, A. Raatgeep, R. H. de Jongste, J. C.2001;Hydrogen peroxide in breath condensate during a common cold351-4Mediators Inflamm106Adult Breath Tests Common Cold/*diagnosis/*metabolism Female Humans Hydrogen Peroxide/*analysis Male Middle Aged Predictive Value of Tests Research Support, Non-U.S. Gov'tDecBACKGROUND: Hydrogen peroxide (H2O2) in exhaled air condensate is elevated in inflammatory disorders of the lower respiratory tract. It is unknown whether viral colds contribute to exhaled H2O2. AIM: To assess exhaled H2O2 during and after a common cold. METHODS: We examined H2O2 in the breath condensate of 20 normal subjects with acute symptoms of a common cold and after recovery 2 weeks later and, similarly, in 10 subjects without infection. H2O2 was measured with a fluorimetric assay. RESULTS: At the time of infection exhaled H2O2 (median, ranges) was 0.20 microM (0.03-1.2 microM), and this decreased to 0.09 microM (< 0.01-0.40 microM) after recovery (p = 0.006). There was no significant difference in lung function (forced vital capacity and forced expiratory volume in 1 sec) during and after colds. In the controls, exhaled H2O2 did not change over a 2-week period. CONCLUSIONS: H2O2 in exhaled air condensate is elevated during a common cold, and returns to normal within 2 weeks of recovery in healthy subjects. Hence, symptomatic upper respiratory tract infection may act as a confounder in studies of H2O2 as a marker of chronic lower airway inflammation.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11817678 !0962-9351 (Print) Journal Article11817678Department of Paediatrics, Division of Paediatric Respiratory Medicine, Erasmus University Medical Center/Sophia Children's Hospital, Rotterdam, The Netherlands. ~?XHunt, J. F. Erwin, E. Palmer, L. Vaughan, J. Malhotra, N. Platts-Mills, T. A. Gaston, B.2002SExpression and activity of pH-regulatory glutaminase in the human airway epithelium101-7Am J Respir Crit Care Med1651Acute Disease Adult Ammonia/analysis/metabolism Analysis of Variance Asthma/*enzymology/immunology/physiopathology Biopsy Blotting, Western Breath Tests Case-Control Studies Cell Survival Gene Expression Regulation/*physiology Glutaminase/analysis/chemistry/*physiology Humans Hydrogen-Ion Concentration Immunohistochemistry Inflammation Interferon Type II/pharmacology/physiology Isomerism RNA, Messenger/analysis Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Respiratory Mucosa/*enzymology Reverse Transcriptase Polymerase Chain Reaction Tumor Necrosis Factor-alpha/pharmacology/physiology Up-Regulation/drug effects/*physiologyJan 18Fluid condensed from the breath of patients with acute asthma is acidic. Several features of asthma pathophysiology can be initiated by exposure of the airway to acid. In renal tubular epithelium, glutaminase produces ammonia to buffer urinary acid excretion. We hypothesized that human airway epithelium could also express glutaminase. Here, we demonstrate that human airway epithelial cells in vitro have biochemical evidence for glutaminase activity and express mRNA for two glutaminase isoforms (KGA and GAC). Glutaminase activity increased in response to acidic stress (media pH 5.8) and was associated with both increased culture medium pH and improved cell survival. In contrast, activity was inhibited by interferon-gamma and tumor necrosis factor-alpha. Glutaminase protein was expressed in the human airway in vivo. Further, ammonia levels in the breath condensate of subjects with acute asthma were low (30 microM [range: 0-233], n = 18, age 23 +/- 2.5 yr) compared with control subjects (327 microM [14-1,220], n = 24, age 24 +/- 2.4 yr, p < 0.001), and correlated with condensate pH (r = 0.58, p < 0.001). These data demonstrate that glutaminase is expressed and active in the human airway epithelium and may be relevant both to the regulation of airway pH and to the pathophysiology of acute asthmatic airway inflammation.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11779738 !1073-449X (Print) Journal Article11779738Division of Pediatric Respiratory Medicine and Division of Allergy, Asthma and Immunology, The University of Virginia Health System, Charlottesville, Virginia 22908, USA.~? Wilson, N.2002,Measurement of airway inflammation in asthma25-32Curr Opin Pulm Med81(Asthma/diagnosis/*pathology/physiopathology/*therapy Biological Markers/analysis Blood Proteins/analysis Breath Tests Eosinophil Granule Proteins Eosinophils/physiology Glucocorticoids/therapeutic use Humans Inflammation Mediators/physiology Nitric Oxide/physiology *Ribonucleases Sputum/cytologyJanChronic airway inflammation is considered responsible for symptoms and disorders of airway function associated with asthma. This process is the target of anti-inflammatory therapy, so a number of standardized, noninvasive techniques have been developed to assess it. More recent approaches include the measurement of exhaled gases and nonvolatile substances in breath condensate. Results from studies using a wide variety of inflammatory markers have shown group differences between patients with asthma and healthy control subjects, but evidence for the diagnostic use of these markers in individual patients is scarce. Similarly, despite many studies demonstrating some correlation between markers of airway inflammation and a measure of disease control, none has yet convincingly shown a place for the use of these markers in an individual with corticosteroid-treated asthma. However, application of these markers continues to further our understanding of the disease process and provides the potential for more appropriate, customized therapy.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11753120 (1070-5287 (Print) Journal Article Review11753120Department of Paediatrics, Royal Brompton and Harefield National Health Service Trust, London, United Kingdom. n.wilson@rbh.nthames.nhs.uk~?Griese, M. Latzin, P. Beck, J.2001TA noninvasive method to collect nasally exhaled air condensate in humans of all ages915-20Eur J Clin Invest3110!Adolescent Adult Air Child Child, Preschool Female Humans Hydrogen Peroxide/analysis/metabolism Hypersensitivity, Immediate/diagnosis/metabolism Infant Infant, Newborn Lung/*metabolism Lung Diseases/diagnosis/metabolism Male Nose *Respiration Respiratory Function Tests/*methods Skin TestsOctBACKGROUND: The analysis of exhaled breath condensate may provide valuable insights into inflammatory and other metabolic processes of the lungs. However, its collection by active exhalation with conventional methods is cumbersome, demands a substantial level of co-operation with high motivation and is very difficult or impossible in children younger than about 4-6 years or in the elderly. A comfortable, noninvasive and efficient method is desirable. DESIGN AND PATIENTS: For collection a high-performance pump connected to a cold trap and nasal prongs were used. The volume of the condensate collected was assessed in 141 children of all ages and five adults. As an example for a low molecular component, H2O2 a marker of oxidative stress, was determined fluorometrically. RESULTS: On average, in healthy children from 4 weeks to 18 years of age, 84.0 (79.4, 87.3) microL min(-1) of nasally exhaled air condensate were collected. The volume obtained was about 45% less in 1-6-year-old children, increased linearly with collection time, and averaged about 20-30% of the exhaled water vapour. The concentration of H2O2 in the healthy children was 0.49 (0.48, 0.61) microM and did not depend on age, the time of the day, family, or personal history of atopy and sex. CONCLUSIONS: The method described is generally applicable, comfortable, noninvasive, safe and efficient and allows the collection of nasally exhaled air condensate for the evaluation of metabolic processes of the lungs.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11737231 !0014-2972 (Print) Journal Article11737231Kinderklinik and Kinderpoliklinik im Dr von Haunerschen Kinderspital, Ludwig-Maximilians-University, Munich, Germany. griese@pk-i.med.uni-muenchen.de <~?^Emelyanov, A. Fedoseev, G. Abulimity, A. Rudinski, K. Fedoulov, A. Karabanov, A. Barnes, P. J.2001JElevated concentrations of exhaled hydrogen peroxide in asthmatic patients1136-9Chest1204Adolescent Adult Airway Obstruction/diagnosis/physiopathology Asthma/*diagnosis/physiopathology Blood Proteins/physiology *Breath Tests Bronchial Hyperreactivity/diagnosis/physiopathology Bronchial Provocation Tests Eosinophil Granule Proteins Female Forced Expiratory Volume/physiology Humans Hydrogen Peroxide/*analysis Male Middle Aged Research Support, Non-U.S. Gov't *Ribonucleases Risk FactorsOctBACKGROUND: Airway inflammation is important in the development and progression of asthma. Activation of inflammatory cells induces a respiratory burst resulting in the production of reactive oxygen species, such as H(2)O(2). The aim of this study was to measure the concentration of H(2)O(2) in exhaled breath condensate and its correlation with airway obstruction, airway hyperresponsiveness, and concentration of eosinophil cationic protein (ECP) in serum in 70 steroid-naive, atopic patients with unstable asthma (20 men; age range, 18 to 62 years) and 17 normal subjects (7 men; age range, 19 to 34 years). METHODS: Exhaled H(2)O(2) was measured using a colorimetric assay, and the concentration of ECP in serum was measured using radioimmunoassay. Airway hyperresponsiveness was expressed as the provocative concentration of inhaled histamine causing a 20% fall in FEV(1) (PC(20)). RESULTS: In patients with asthma, the mean H(2)O(2) concentration was significantly elevated compared to values in normal subjects: 0.127 +/- 0.083 mol/L vs 0.024 +/- 0.016 mol/L (p < 0.001). There was a significant correlation among H(2)O(2) concentration, FEV(1), PC(20), and ECP in serum. CONCLUSION: We conclude that exhaled H(2)O(2) is significantly elevated in asthmatic patients. This is correlated with disease severity and indirect markers of airway inflammation. Measurement of exhaled H(2)O(2) may be useful to assess airway inflammation and oxidative stress in asthmatic patients.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11591550 !0012-3692 (Print) Journal Article11591550cHospital Therapeutic Clinic, Pavlov Medical University, St. Petersburg, Russia. emelav@netscape.net~?DKhasina, M. A. Vas'kova, N. A. Soboleva, I. V. Solomiannikova, T. F.2001r[Informative value of bronchoalveolar lavage and expired air vapor condensate in the diagnosis of acute pneumonia]25-7 Probl Tuberk5Acute Disease Adolescent Adult Breath Tests Bronchoalveolar Lavage Fluid Comparative Study Humans Male Pneumonia/*diagnosis/metabolism/physiopathology Pulmonary Surfactants/metabolismfhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11588954 !0032-9533 (Print) Journal Article11588954tInformativnost' bronkhoal'veoliarnogo smyva i kondensata parov vydykhaemogo vozdukha v diagnostike ostroi pnevmonii. ~?DFerreira, I. M. Hazari, M. S. Gutierrez, C. Zamel, N. Chapman, K. R.2001Exhaled nitric oxide and hydrogen peroxide in patients with chronic obstructive pulmonary disease: effects of inhaled beclomethasone1012-5Am J Respir Crit Care Med1646Administration, Topical Analysis of Variance Anti-Inflammatory Agents/*administration & dosage/*pharmacology Beclomethasone/*administration & dosage/*pharmacology Breath Tests Chemiluminescent Measurements Comparative Study Cross-Over Studies Double-Blind Method Female Glucocorticoids Humans Hydrogen Peroxide/*analysis Male Nitric Oxide/*analysis Placebos Prospective Studies Pulmonary Disease, Chronic Obstructive/*drug therapy/physiopathology Research Support, Non-U.S. Gov't *Respiratory Therapy Spectrophotometry SpirometrySep 15There is controversy about the role of inhaled corticosteroids in chronic obstructive pulmonary disease (COPD). Although they appear to have little impact on airways obstruction or its progression, their use may reduce the frequency and/or severity of exacerbations in a subset of patients. We undertook the following study to determine the impact of inhaled corticosteroid on two noninvasive markers of airways inflammation. We assigned 20 stable nonsmoking patients with COPD in random, double-blind crossover fashion to two 2-wk treatment periods with inhaled beclomethasone 500 microg twice daily or matching placebo, followed by a 2-wk washout period. We measured exhaled nitric oxide (ENO), breath condensate H(2)O(2), and flow volume spirometry at weekly intervals. Median baseline ENO was 26.2 (19.3 to 54.8) ppb and fell significantly following 1 and 2 wk of beclomethasone (-10.6 ppb, p = 0.002, and -6.3 ppb, p = 0.013, respectively) but was unchanged by placebo inhalation. Breath condensate H(2)O(2) levels did not change significantly with inhaled beclomethasone or placebo. Although there were no significant changes in FEV(1) with BDP therapy, there was a moderate inverse correlation between changes in ENO and changes in FEV(1) (r -0.50). We conclude that inhaled beclomethasone reduces ENO levels in stable nonsmoking patients with COPD, a finding compatible with an antiinflammatory mechanism of action.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11587988 L1073-449X (Print) Clinical Trial Journal Article Randomized Controlled Trial11587988tAsthma Centre, University of Toronto, Toronto Western Hospital, University Health Network, Toronto, Ontario, Canada.~?GMutlu, G. M. Garey, K. W. Robbins, R. A. Danziger, L. H. Rubinstein, I.2001>Collection and analysis of exhaled breath condensate in humans731-7Am J Respir Crit Care Med1645 Asthma/metabolism Breath Tests/*methods Forecasting Humans Pulmonary Disease, Chronic Obstructive/metabolism Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Respiratory Distress Syndrome, Adult/metabolism Smoking/metabolism Specimen HandlingSep 1fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11549524 (1073-449X (Print) Journal Article Review11549524Pulmonary and Critical Care Medicine, Evanston-Northwestern Healthcare and Northwestern University Medical School, Evanston, Illinois, USA. O~?QGessner, C. Kuhn, H. Seyfarth, H. J. Pankau, H. Winkler, J. Schauer, J. Wirtz, H.2001,Factors influencing breath condensate volume414-9 Pneumologie559 Asthma/*diagnosis Breath Tests/*instrumentation Humans Humidity Lung Volume Measurements Nitric Oxide/analysis Proteins/*analysis Pulmonary Disease, Chronic Obstructive/*diagnosis Reference Values Research Support, Non-U.S. Gov't Sensitivity and Specificity TemperatureSepAnalysis of breath condensate (BC) has received interest recently due to the need for easy and repetitive monitoring or airway and pulmonary disease. While many authors have used custom built systems, commercial systems are now available and will probably be used more widely. Early studies of markers and mediators in BC have reported concentrations following varying periods of sampling time. However, factors that influence the generation of BC have not been analysed and it is unclear whether breathing rate, tidal volume, lung function, body weight, height or age influence the amount of BC collected. We therefore studied the influence of these factors on breath condensate volume and breath condensate urea and protein concentrations in 22 healthy volunteers and 23 COPD patients. A strong correlation of total respired volume and breath condensate volume was observed for both groups (volunteers: r=0.952, p < 0.0001, COPD: r=0.883, p < 0.001) while no significant correlation existed for breath condensate volume and TLC, RV, Vc, FEV1, R(tot), height or body weight, As long as ventilation remained fairly constant, breath condensate volume increased linearly with time. The fraction of breath condensate extracted from total vapour contained in the exhalate was estimated by measuring relative atmospheric humidity before and after the collecting tube. The amount calculated by the change in temperature and saturation corresponded closely to the amount actually collected. We conclude from these results that breath condensate volume is primarily dependent on V(E), and does not seem to depend on lung function parameters. For standardisation it is suggested to report breath condensate measurements per volume respired. Both, urea and protein are present in measurable quantities in breath condensate and protein as well as BCV may be helpful denominators for comparison with e.g. cytokines in lung disease.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11536064 !0934-8387 (Print) Journal Article11536064hMedizinische Klinik und Poliklinik I, Universitatsklinikum Leipzig, Germany. gesc@medizin.uni-leipzig.de ~?GGanas, K. Loukides, S. Papatheodorou, G. Panagou, P. Kalogeropoulos, N.2001JTotal nitrite/nitrate in expired breath condensate of patients with asthma649-54 Respir Med958AAdministration, Topical Adrenergic beta-Antagonists/therapeutic use Adult Anti-Inflammatory Agents/therapeutic use Asthma/drug therapy/*metabolism Biological Markers/analysis Breath Tests Case-Control Studies Glucocorticoids Humans Hydrogen Peroxide/analysis Nitrates/*analysis Nitrites/*analysis Oxidative Stress SmokingAugProduction of nitric oxide (NO) is generally increased during inflammatory diseases including asthma. The eventual fate of NO is oxidation to nitrite (NO2) and nitrate (NO3), both of which are end-products of NO metabolism. Hydrogen Peroxide (H2O2) is increased in exhaled breath condensate of asthmatic subjects and may be used as a non-invasive marker of oxidative stress. NO has in some cases been shown to attenuate oxidant-induced lung injury. Total NO2/NO3 concentration and H2O2 levels were measured in expired breath condensate in 50 clinically stable asthmatics [all males, all atopics, mean age 22 (3) SD yrs, forced expiratory volume in 1 sec (FEV1) 91 (10)% predicted, PD20 to histamine 0.262 (0.16) mg 20 on inhaled steroids, 20 smokers, all steroid-naive] and in 10 normal, non-atopic subjects [all males, age 23 (4) yrs, FEV1 101 (14)% predicted, PD20 to histamine 1.3 (0.55) mg]. NO2/NO3 levels were significantly higher in patients with asthma than in normal subjects (1.08, 95% CI 0.86-1.3 microM vs. 0.6; 95% CI 0.46-0.8, P < 0.001). Patients who were on inhaled steroids had significantly ower values compared to steroid-naive (0.71, 95% CI 0.55-0.87 microM vs. 133, 95% CI 1-1.65 microM, P < 0.001). Similar results were observed between smokers and non-smokers (1.11, 95% CI 0.74-1.47 microM vs. 1.77, 95% CI 1.1-24 microM, P < 0.0001).There was a significant positive correlation between NO2/NO3 levels and H2O2 concentration in expired breath condensate (r = 0.48, P < 0.0001). No correlation was observed between NO2/NO3 levels, airway obstruction and bronchial hyper-reactivity as assessed by PD20 to histamine. Total NO2/NO3 levels in expired breath condensate are raised in patients with stable asthma and are significantly related to oxidative stress as assessed by H2O2 concentration. Measurement of expired breath NO2/NO3 and H2O2 levels may be clinically useful in the management of oxidation and inflammation mediated lung injury.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11530952 !0954-6111 (Print) Journal Article11530952CThoracic Medicine Department, Athens Army General Hospital, Greece.~?Horvath, I. MacNee, W. Kelly, F. J. Dekhuijzen, P. N. Phillips, M. Doring, G. Choi, A. M. Yamaya, M. Bach, F. H. Willis, D. Donnelly, L. E. Chung, K. F. Barnes, P. J.2001"Haemoxygenase-1 induction and exhaled markers of oxidative stress in lung diseases", summary of the ERS Research Seminar in Budapest, Hungary, September, 1999420-30 Eur Respir J182Antioxidants/pharmacology Biological Markers/analysis Breath Tests Carbon Monoxide/metabolism Enzyme Induction/drug effects Heme Oxygenase (Decyclizing)/*biosynthesis Humans Lung Diseases/enzymology/*metabolism *Oxidative Stress/drug effects Research Support, Non-U.S. Gov'tAugXIn recent years, there has been increasing interest in noninvasive monitoring of airway inflammation and oxidative stress. Several volatile and nonvolatile substances can be measured in exhaled breath and have been suggested as potential biomarkers of these events. Exhaled gases, including carbon monoxide (CO), alkanes (ethane, pentane), and substances measured in breath condensate, such as hydrogen peroxide (H2O2) and isoprostanes were all suggested as potential markers of oxidative stress in the lung. A European Respiratory Society (ERS) International Research Seminar entitled "Haemoxygenase-1 induction and exhaled markers of oxidative stress in lung diseases" was organized by the Airway Regulation and Provocation Group of the Clinical Allergy and Immunology Assembly in Budapest, Hungary in September, 1999 to integrate the latest knowledge on these issues and accelerate further improvement in this area. During this 2-day event several issues were raised about: the use and standardization of measurements in exhaled breath; problems of measuring expired H2O2 and other mediators in breath condensate; role and regulation of haemoxygenase (HO)-1 in the lung; and conditions and factors influencing exhaled CO. This report is a summary of the main presentations at the seminar, together with the current areas of research in this rapidly expanding field.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11529303 0903-1936 (Print) Congresses11529303WNational Koranyi Institute for Pulmonology, Dept of Pathophysiology, Budapest, Hungary.~?YZappacosta, B. Persichilli, S. Mormile, F. Minucci, A. Russo, A. Giardina, B. De Sole, P.2001TA fast chemiluminescent method for H(2)O(2) measurement in exhaled breath condensate187-91Clin Chim Acta3102Adult *Chemiluminescent Measurements Humans Hydrogen Peroxide/*analysis Middle Aged Pulmonary Alveoli/metabolism Reference Values *Respiration Sensitivity and Specificity Smoking/metabolismAug 20BACKGROUND: Breath condensate can give useful information on volatile compounds produced at alveolar level. Actual concentration of H(2)O(2) in breath condensate is dependent on its production at alveolar level and on the efficacy of the detoxifying systems, catalase, glutathione peroxidase, etc. METHODS: In the present paper, a simple chemiluminescent method for the determination of the H(2)O(2) collected in exhaled breath is shown and data of both smokers and nonsmokers volunteers are presented. RESULTS: The chemiluminescent response is linear up to 100 micromol/l H(2)O(2). The analytical sensitivity is about 0.01 micromol/l. Most of the nonsmokers have a H(2)O(2) content lower than 0.05 micromol/l, while smokers have a content ranging from 0.1 to 0.6 micromol/l.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11498084 !0009-8981 (Print) Journal Article11498084{Istituto di Chimica e Chimica Clinica and CNR Centro Chimica dei Recettori, Rome, Italy. b.zappacosta@uniserv.ccr.rm.cnr.it~?$Khyshiktuev, B. S. Maksimenia, M. V.2001y[Diagnostic value of the determination of fatty acid spectrum of expired air condensate in the workers of coal stripping]38-40 Gig Sanit2$Adult Age Factors *Breath Tests *Coal *Coal Mining Comparative Study Dust/*adverse effects English Abstract Fatty Acids/*analysis Female Humans Lipid Peroxidation Male Middle Aged Occupational Diseases/*etiology *Occupational Exposure Respiratory Tract Diseases/*etiology Siberia Time FactorsMar-AprThe fatty acid spectrum of an expired air condensate was studied in the workers of the Kharanor coal stripping and in the dwellers of Chita (a control group). Regularities in the changes of the expired air fatty acid profile were assessed by the degree of exposure to coal dust and by the length of service. The findings indicated the relationship of profile changes with the degree of exposure to occupational noxious agents: the proportion of saturated fatty acids decreases much more significantly in workers who were directly exposed to the dust than in those were indirectly done. It is concluded that the fatty acid composition of an expired air condensate adequately reflects the changes occurring in the respiratory system upon exposure to dust.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11494488 !0016-9900 (Print) Journal Article11494488Diagnosticheskie znacheniia opredeleniia zhirno-kislotnogo spektra kondensata vydykhaemogo vozdukha u rabochikh ugol'nogo razreza. Q~?[Linnane, S. J. Thin, A. G. Keatings, V. M. Moynihan, J. B. McLoughlin, P. FitzGerald, M. X.2001QGlucocorticoid treatment reduces exhaled nitric oxide in cystic fibrosis patients1267-70 Eur Respir J176Adult *Breath Tests Cross-Over Studies Cystic Fibrosis/*drug therapy Double-Blind Method Female Humans Lung Volume Measurements Male Nitric Oxide/*analysis Prednisolone/*therapeutic use Research Support, Non-U.S. Gov't SpirometryJunCIn cystic fibrosis (CF), low concentrations of exhaled nitric oxide (NO) and reduced expression of inducible nitric oxide synthase (iNOS) in airway epithelium have been reported. However, abundant iNOS expression has been found in the subepithelial tissues and elevated concentrations of NO metabolites in breath condensate and sputum. These conflicting results may be explained by increased scavenging of NO by superoxide radicals, resulting in rapid conversion to peroxynitrite, so that only a small proportion of the NO produced in the lung tissue reaches the airway lumen. If iNOS were active in the CF lung, exhaled NO would be further reduced by glucocorticoid treatment. CF patients (n = 13) were recruited to a double-blind, placebo-controlled study with crossover. Treatment comprised prednisolone or placebo for 5 days with a 9 day washout. After each treatment, exhaled NO was measured, spirometry performed and blood collected for measurement of serum nitrogen dioxide/nitrous oxide (NO2/NO3). Ten patients (8 male) completed the study. Following prednisolone treatment (mean +/- SD) exhaled NO concentration (3.1 +/- 1.6 parts per billion (ppb)) was significantly reduced versus placebo treatment (4.9 +/- 4.2 ppb; p<0.05, Wilcoxon signed-rank test). Spirometric indices and serum NO2/NO3 concentration were unchanged. These findings support the hypothesis that glucocorticoids suppress nitric oxide production in cystic fibrosis airways by reducing inducible nitric oxide synthase expression or by inhibiting recruitment of neutrophils, cells which express inducible nitric oxide synthase.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11491175 L0903-1936 (Print) Clinical Trial Journal Article Randomized Controlled Trial11491175qDept of Medicine and Therapeutics, Conway Institute of Biomolecular Medicine, University College Dublin, Ireland.~?aBalint, B. Kharitonov, S. A. Hanazawa, T. Donnelly, L. E. Shah, P. L. Hodson, M. E. Barnes, P. J.2001GIncreased nitrotyrosine in exhaled breath condensate in cystic fibrosis1201-7 Eur Respir J176Adult *Breath Tests Cystic Fibrosis/*diagnosis Female Humans Lung Volume Measurements Male Oxidative Stress Reactive Nitrogen Species/analysis Reference Values Research Support, Non-U.S. Gov't Tyrosine/*analogs & derivatives/*analysisJunExhaled nitric oxide (ENO), a marker of inflammation in airway diseases is decreased in cystic fibrosis (CF) patients, perhaps because nitric oxide (NO) is metabolized to oxidative end-products. A stable product, 3-nitrotyrosine, may indicate local formation of reactive nitrogen species. Whether NO metabolites in exhaled breath condensate may be increased in CF patients was investigated. The fractional concentration of ENO (Feno), nitrotyrosine and oxides of nitrogen in exhaled breath condensate from 36 stable CF patients were compared to 14 normal subjects using an enzyme immunoassay and fluorescence assay. Nitrotyrosine levels in breath condensate were increased significantly in stable CF patients, compared with normal subjects (25.3 +/- 1.5 versus 6.3 +/- 0.8 ng x mL(-1), p<0.0001). There was an inverse correlation between the levels of nitrotyrosine and the severity of lung disease. Feno levels were significantly lower in CF patients than in normal subjects (4.4 +/- 0.3 versus 5.6 +/- 0.4 (parts per billion), p<0.05). No correlation was found between nitrotyrosine and Feno levels in CF. There was no significant difference in the levels of nitrite and nitrate between CF patients and normals. The elevation in nitrotyrosine may reflect increased formation of reactive nitrogen species such as peroxynitrite or direct nitration by granulocyte peroxidases, indicating increased oxidative stress in airways of cystic fibrosis patients.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11491165 !0903-1936 (Print) Journal Article11491165tDept of Thoracic Medicine, Imperial College School of Medicine at the National Heart and Lung Institute, London, UK. ~?Kasielski, M. Nowak, D.2001Long-term administration of N-acetylcysteine decreases hydrogen peroxide exhalation in subjects with chronic obstructive pulmonary disease448-56 Respir Med956Acetylcysteine/*therapeutic use Adult Aged Analysis of Variance Breath Tests Double-Blind Method Female Free Radical Scavengers/*therapeutic use Humans Hydrogen Peroxide/*metabolism Lipid Peroxidation/drug effects Lung Diseases, Obstructive/*drug therapy/metabolism Male Middle Aged Research Support, Non-U.S. Gov't Spectrometry, Fluorescence Statistics, Nonparametric Thiobarbituric Acid Reactive Substances/analysis Treatment OutcomeJun Patients with chronic obstructive pulmonary disease (COPD) exhale more hydrogen peroxide (H2O2) and lipid peroxidation products than healthy subjects. This may reflect oxidative stress in the airways that plays important role in the development and progression of COPD. N-acetylcysteine (NAC), a mucolytic drug, possesses antioxidant properties as it is a precursor of reduced glutathione that together with glutathione peroxidase may decompose H2O2 and lipid peroxides. We aimed to determine the effect of NAC, 600 mg effervescent tablets (Fluimucil), once a day for 12 months, and placebo on the concentration of H2O2 and thiobarbituric acid reactive substances (TBARs) in expired breath condensate and serum levels of two lipid peroxidation products (TBARs, lipid peroxides) in patients with COPD. The study was performed as a double-blind, double-dummy comparison between active drug and placebo in two parallel groups. Forty-four outpatients with stable COPD (22 in the NAC group and 22 in the placebo group) completed the study. Specimens of expired breath condensate and serum were collected at the randomization visit and then every 3 months over 1 year. The concentration of TBARs and H2O2 in expired breath condensate was measured spectrofluorimetrically by the thiobarbituric acid and homovanillic acid methods, respectively. Serum levels of lipid peroxides were determined spectrophotometrically after extraction with butanol and pyridine. Initially, H2O2 exhalation did not differ between the placebo and NAC groups up to 6 months of treatment. After this the significant differences were observed. After 9 and 12 months of treatment NAC group exhaled 2.3-fold (0.17+/-0.33 microM vs. 041+/-0.26 microM, P<0.04) [median 0.01 microM, quartile range (qr)=0.22 vs. median 0.15 microM, qr =0.43] and 2.6-fold (0.15+/-0.23 microM vs. 0.40+/-0.25 microN, P<0.05) median = 0.00 microM, qr = 0.23 vs. median = 0.36 microM, qr = 0.51] less H2O2 than placebo receivers, respectively. No significant effect of NAC administration on TBARs exhalation and serum levels of TBARs and lipid peroxides were noted over the whole treatment period. Also no significant associations between exhaled H2O2 and concentrations of lipid peroxidation products were noted in both treatment groups at any time-point. These results indicate that long-term oral administration of NAC attenuates H2O2 formation in the airways of COPD subjects and prove anti-oxidant action of drug. However, further studies are necessary to estimate the clinical significance of this finding.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11421501 ^0954-6111 (Print) Clinical Trial Journal Article Multicenter Study Randomized Controlled Trial11421501dCentre of Medical Education, Practical Clinical Training Centre, Medical University of Lodz, Poland. ~?Nevzorova, V. A. Prosekova, E. V. Gel'tser, B. I. Luk'ianov, P. A. Shestovskaia, T. N. Maistrovskaia Iu, V. Arzamastseva, A. I.2001z[Changes in biochemical inflammation markers in evaluation of the effectiveness of basic chemotherapy in bronchial asthma]24-7Ter Arkh733PAdolescent Adult Age Factors Androstadienes/administration & dosage/therapeutic use Anti-Asthmatic Agents/administration & dosage/*therapeutic use Anti-Inflammatory Agents/administration & dosage/*therapeutic use Asthma/diagnosis/*drug therapy/metabolism Biological Markers Breath Tests Child Child, Preschool Comparative Study Cromolyn Sodium/administration & dosage/therapeutic use English Abstract Female Humans Inflammation/diagnosis/metabolism Ketotifen/administration & dosage/therapeutic use Male Nedocromil/administration & dosage/therapeutic use Nitric Oxide/*metabolism Time Factors~AIM: To analyse informative value of monitoring of NO metabolites concentration in condensate of expired air vapor (EAVC) for definition of bronchial asthma (BA) severity and antiinflammatory effect of basic therapy. MATERIAL AND METHODS: NO metabolites concentration in EAVC was measured with Grace's reagent in 76 adults and 180 children in the course of basic therapy with flixotide, tailed, ketotiphen with intal. RESULTS: NO metabolites occurred in high concentrations in EAVC of both children and adults. These concentrations were the highest in attacks and in severe BA and were reduced by basic therapy. This effect differed with the drug and its dose, e.g. flixotide and tailed given to children for a month reduced NO metabolites close to concentrations observed in healthy subjects. Ketotiphen and intal for 6 months failed to low NO metabolites significantly. CONCLUSION: NO secretion monitoring is sensitive in assessment of respiratory inflammation in BA and is informative in assessment of effectiveness of the basic therapy. In moderate BA children on flixotide and tailed were the first to achieve normal NO secretion in the airways.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11417175 !0040-3660 (Print) Journal Article11417175wDinamika biokhimicheskikh markerov vospaleniia v otsenke effektivnosti bazisnoi farmakoterapii pri bronkhial'noi astme.<~?Nikitin, A. V. Zuikova, A. A.2001[Clinical effectiveness of superoxide inhalations and their effects on crystalline structure and antiradical defense of blood serum and expired air condensate in patients with bronchial asthma]20-3Ter Arkh733Z*Antioxidants Asthma/*drug therapy Breath Tests Ceruloplasmin/*analysis Comparative Study Crystallization English Abstract Free Radical Scavengers/*blood Glutathione Reductase/*blood Humans Lipid Peroxidation Malondialdehyde/blood Oxidative Stress *Respiratory Therapy Superoxide Dismutase/*blood Superoxides/*administration & dosage Time FactorsHAIM: To raise efficiency of combined treatment of bronchial asthma (BA) by application of intranasal inhalations of gaseous superoxide (GSO) basing on clinicolaboratory, functional and biochemical indices. MATERIAL AND METHODS: The study entered 20 healthy subjects and 57 BA patients. All of the patients received conventional treatment, but 30 of them additionally inhaled GSO. The effect of inhalations was assessed by parameters of LPO products, enzymatic and nonenzymatic antiradical defense (ARD) in the blood and expired air condensate (EAC), morphometry of EAC crystalline structures. RESULTS: GSO intranasal inhalations improve treatment of BA as it produces positive effect on endogenic oxidative stress, LPO, enzymatic and nonenzymatic ARD, blood and EAC tesigraphy. CONCLUSION: Antiinflammatory effect of GSO in BA is suggested.fhttp://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=11417174 00040-3660 (Print) Clinical Trial Journal Article11417174Klinicheskaia effektivnost' ingaliatsii superoksida i ikh vliianie na kristallicheskuiu strukturu i sistemu antiradikal'noi zashchity syvorotki krovi i kondensata vydykhaemogo vozdukha u bol'nykh bronkhial'noi astmoi. ~?GBalint, B. Donnelly, L. E. Hanazawa, T. Kharitonov, S. A. Barnes, P. J.2001_Increased nitric oxide metabolites in exhaled breath condensate after exposure to tobacco smoke456-61Thorax566Adult Analysis of Variance Breath Tests/methods Carbon Monoxide/metabolism Female Humans Male Nitrates/metabolism Nitric Oxide/*metabolism Nitrites/metabolism Nitroso Compounds/metabolism Oxidative Stress Research Support, Non-U.S. Gov't Smoking/adverse effects/*metabolismJunBACKGROUND: Cigarette smoking reduces the level of exhaled nitric oxide (NO) in healthy subjects, although the mechanism is unclear. NO is a highly reactive molecule which can be oxidised or complexed with other biomolecules, depending on the microenvironment. The stable oxidation end products of NO metabolism are nitrite and nitrate. This study investigated the effect of smoking on NO metabolites in exhaled breath condensate. METHODS: Fifteen healthy current smokers were recruited together with 14 healthy non-smokers. Measurement of exhaled NO, lung function, and collection of exhaled breath condensate were performed. Nitrite, nitrite + nitrate, S-nitrosothiols, and nitrotyrosine levels were measured. The effect of inhaling two cigarettes in smokers was also evaluated. The mean level of exhaled NO in smokers was significantly lower than in non-smokers (4.3 (0.3) ppb v 5.5 (0.5) ppb, p<0.05). RESULTS: There was no difference in the levels of nitrite, nitrite + nitrate, S-nitrosothiols, and nitrotyrosine in the exhaled breath condensate at the baseline visit between smokers and non-smoke