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ABSTRACT |
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TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
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Nitrosothiols (RS-NOs) are formed by interaction of nitric oxide
(NO) with glutathione and may limit the detrimental effect of NO.
Because NO generation is increased in airway inflammation, we
have measured RS-NOs in exhaled breath condensate in patients with asthma, cystic fibrosis, or chronic obstructive pulmonary disease (COPD). We also measured exhaled NO and nitrite (NO2
) in
the same subjects. RS-NOs were detectable in exhaled breath condensate of all subjects. RS-NOs were higher in subjects with severe
asthma (0.81 ± 0.06 µM) when compared with normal control subjects (0.11 ± 0.02 µM, p < 0.01) and with subjects with mild asthma (0.08 ± 0.01 µM, p < 0.01). Elevated RS-NOs values were also found in patients with cystic fibrosis (0.35 ± 0.07 µM, p < 0.01), in those with COPD (0.24 ± 0.04 µM, p < 0.01) and in smokers (0.46 ± 0.09 µM, p < 0.01). In current smokers there was a correlation (r = 0.8, p < 0.05) between RS-NOs values and smoking history (pack/year). We also found elevated concentrations of NO2 in patients with severe asthma, cystic fibrosis, or COPD, but not in smokers or patients with mild asthma. This suggests that exhaled NO2 is less sensitive than exhaled RS-NOs. This study has
shown that RS-NOs are detectable in exhaled breath condensate
of healthy subjects and are increased in patients with inflammatory airway diseases. As RS-NOs concentrations in exhaled breath
condensate vary in the different airway diseases and increase with
the severity of asthma, their measurement may have clinical relevance as a noninvasive biomarker of nitrosative stress.
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INTRODUCTION |
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TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
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Endogenous nitric oxide (NO) plays an important role in the physiological regulation of the airways and is implicated in the pathophysiology of airway diseases (1). NO is formed endogenously in human lung by NO synthase (NOS), of which three isoforms have been identified in human airways (2). Constitutive isoforms of NOS (cNOS) are expressed predominantly in pulmonary vasculature and in airway nerves (2). The inducible isoform of NOS (iNOS), which is expressed in both normal and pathologic lung tissues (2, 3), is induced by inflammatory stimuli, including proinflammatory cytokines and bacterial products (3). Induction of both iNOS and cNOS during airway inflammation may result in increased concentrations of exhaled NO, as demonstrated in inflammatory airway diseases such as asthma (4), bronchiectasis (4), and upper respiratory tract infection (4). In addition, with regard to asthma, it has been recently proposed that airway acidification may explain increased levels of exhaled NO (5). Exhaled NO is currently considered as a marker of airway inflammation (6).
NO is a free radical, by virtue of its unpaired electron. Under physiologic conditions, NO is unstable, reacting with oxygen to form oxides of nitrogen (NOx), such nitrite (NO2) and
nitrate (NO3), and with superoxide anion to form the potent
oxidant peroxynitrite (ONOO) (7). ONOO reacts with a
wide variety of compounds, including DNA, cellular lipids, and
sulphydryl groups on proteins and can thus promote nitrosative stress (8).
Little is known about the antioxidant response of the respiratory tract to chronic inflammation in human airways. Pulmonary antioxidant defenses are widely distributed and include both enzymatic and nonenzymatic systems (11). The major enzymatic antioxidants are based on thiols (11), which are classified according to their molecular weight. Low molecular weight thiols include glutathione (GSH), cysteine, homocysteine, albumin, and N-acetylcysteine; high molecular weight thiols include capropril and penicillamine (12); GSH is the predominant endogenous thiol in human lung (13). NO and NO metabolites interact readily with the thiol groups of thiols to produce S-nitrosothiols (RS-NOs) (14). The general formula of RS-NOs is R-S-N = O, where R is an alkyl or aryl group of thiols.
As NO itself is short-lived in vivo and its rapid reaction with superoxide results in its inactivation and in the generation of NOx, the formation of RS-NOs serve to stabilize NO in a form that is noncytotoxic (7). However, RS-NOs do not simply provide a means of protecting the NO group from reactions with superoxide, rather they endow NO with enhanced bioactivity. In fact, RS-NOs are biologically active as vasodilators and bronchodilators (13). Therefore, RS-NOs rather than NO are the major products of NOS (15), representing by far the largest pool of NO bioactivity in both the lung and the blood (13, 15).
Increased airway RS-NOs values have been measured in BAL fluid of subjects with infective pneumonia and in subjects with transplanted lung, suggesting that inflammation and/or immune activation may be important stimulants of RS-NOs generation (13).
Exhaled breath condensate (EBC), which is obtained by freezing exhaled air under conditions of spontaneous breathing, contains nonvolatile substances from the lower airways (18). Analysis of EBC may provide a noninvasive means of monitoring airway inflammation and cellular metabolism of the lung (19). We hypothesized that soluble RS-NOs are formed in aerosols in the breath and are then detectable in EBC.
The aim of the present study was to investigate whether RS-NOs could be detected in EBC of a spectrum of patients with inflammatory airway diseases and to compare RS-NOs values in these patients with those in healthy smoking and nonsmoking subjects.
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METHODS |
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TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
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Subject Characteristics
Six groups of subjects were recruited for this study: normal non-smoking control subjects (n = 10); smoking healthy subjects (n = 7); subjects with mild asthma (n = 9); subjects with severe asthma (n = 8); and those with cystic fibrosis (CF) (n = 10) or COPD (n = 7) (Table 1). All the patients with asthma, COPD or CF and all the normal subjects and smokers were recruited and examined at the Department of Thoracic Medicine/Brompton Hospital, London, and the study had Ethics Committee approval.
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Normal control subjects had no history of respiratory diseases and had normal lung function tests, and they formed the control group. Smokers (11.3 ± 1.8 pack/years) had normal lung function tests and were free of symptoms at the time the study.
All asthmatic conformed to the diagnostic criteria of the Global
Initiative for Asthma (20). Patients with mild intermittent asthma had
occasional symptoms of variable wheeze and dyspnea, controlled by
2-agonists alone, and none had suffered an asthma exacerbation
within the month preceding the study. All subjects demonstrated a > 15% improvement in their FEV1 after receiving 200 µg of nebulized
salbutamol and airway hyperresponsiveness to methacholine, with a
provocative concentration of methacholine producing a 20% fall in
FEV1 (PC20), < 8 mg/ml. None of the subjects with mild asthma studied had received inhaled or orally administered steroids in the preceding 3 mo. Patients with severe persistent asthma were patients seen
for severe, oral-steroid-dependent asthma with frequent hospitalization and emergency visits. They had severe persistent asthma symptoms despite chronic oral steroid treatment.
Nonsmoking patients with CF attending an outpatient clinic for a scheduled visit were tested. None of these patients presented with exacerbation of their respiratory symptoms at the time of the study.
All patients with COPD complained of morning cough with mucoid sputum production on most days of the month, for at least 3 mo per year during at least the previous 2 yr.
They met the following criteria: (1) stable airflow limitation with
FEV1 
80% of predicted value; (2) no improvement in FEV1 of
more than 15% after inhalation of 200 µg of salbutamol; (3) no exacerbation of their disease defined as a change in quality and quantity of
sputum with increased dyspnea in the previous month. All patients
with COPD were ex-smokers (stopped smoking 9.4 ± 1.3 yr).
Study Design
Subjects' details were obtained and then baseline spirometry and exhaled NO were measured, followed by collection of EBC. Bronchial reactivity was tested in the group with mild asthma the day before the study. Current smokers were asked to refrain from smoking at least 8 h before the EBC maneuver.
Pulmonary Function Tests
FEV1 was measured with a dry spirometer (Vitalograph, Buckingham, UK). The best value of the three maneuvers was expressed as a percentage of the predicted value.
Airway responsiveness was measured by methacholine challenge with doubling concentrations of methacholine (0.06 to 32 mg/ml) delivered by dosimeter11 (Mefar, Bovezzo, Italy) with an output of 10 µl per inhalation. The aerosols were inhaled at tidal breathing with the subject wearing a noseclip. Five inhalations of each concentration were administered (inhalation time, 1 s; breathholding time, 6 s). FEV1 was measured 2 min after the last inhalation until there was a fall in FEV1 of 20% (PC20) when compared with the control inhalation (0.9% saline solution) or until the maximal concentration was inhaled.
Exhaled NO
Exhaled NO was measured using a chemiluminescence analyzer (Model LR 2000; Logan Research, Rochester, Kent, UK) sensitive to NO from 1 to 500 by volume parts per billion (ppb), and with a resolution of 0.3 ppb and response time of < 0.5 s. Measurements were made after inhalation to total lung capacity with slow exhalation through a wide-bore Teflon tube to residual volume.
According to European guidelines (4) an expiratory pressure of 3 (0.4) mm Hg was maintained by a visual display of expiratory flow, thus excluding nasal NO contamination. NO sampling by the analyzer was via a side arm with a constant flow rate of 250 ml/min. NO measurement was made at the end of exhalation when both NO and CO2 levels had achieved a stable plateau phase, representing alveolar sampling.
Exhaled Breath Condensate Collection
EBC was collected, as previously described (19) by using a glass-condensing device, with an inner glass chamber, which contained ice and was suspended in a larger chamber. Condensate was formed on the outside surface of the inside glass that was separated from ambient air.
After rinsing their mouths, each subject breathed tidally through a mouthpiece connected to the inlet for 15 min while wearing a noseclip. The mouthpiece was also used as a saliva trap.
Approximately 0.5 ml of EBC was collected and stored at 
70° C. Samples were frozen on liquid nitrogen immediately after collection and measured in 15 d without loss of activity.
RS-NOs Measurements
RS-NOs were measured using a commercially available colorimetric
assay kit (Oxonon, Emeryville, CA). The assay is based upon the classic reaction of Saville and Griess (21, 22). In essence, a cleavage reaction breaks the S-N bond of RS-NOs releasing NO, which oxides rapidly to NO2. NO2 is then detected colorimetrically using the Griess
reaction. Briefly, 200 µl of EBC were used for each assay, and 50 µL
of Griess 1 were added, followed by 50 µL of Griess 2. The product was measured spectrophotometrically (Model AR 8003; Labtech Int. Ltd., Uckfield, UK) at 540 nm. A standard curve of nitrosogluthatione (GS-NO) was performed for each assay. The detection limit of the kit
is 0.025 µM. Levels of RS-NOs were determined by interpolation from known standard curve, and were expressed as µM concentration. All the samples were run in duplicate, and mean values were
used for subsequent analysis.
Statistical Analysis
Data are expressed as mean ± SEM. Differences between groups were analyzed with the Mann-Whitney rank test. Spearman's test was used for correlation analysis. A p value less than 0.05 was considered significant.
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RESULTS |
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TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
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Exhaled RS-NOs
All of the subjects performed the EBC maneuver without discomfort, and RS-NOs were detectable in all subjects (Figure
1). No correlation was found between RS-NOs values and age
in any of the subjects (r = 0.1, p = 0.3) or in each subgroup:
controls (r = 0.2, p = 0.2), asthma (r = 0.2, p = 0.3), CF (r = 0.1, p = 0.6), COPD (r = 0.4, p = 0.2).
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RS-NOs were higher in subjects with severe asthma (0.81 ± 0.06 µM) than in control subjects (0.11 ± 0.02 µM, p < 0.01), or in patients with mild asthma (0.08 ± 0.01 µM, p < 0.01). RS-NOs values in subjects with mild asthma were not different from those of the control subjects.
Compared with control subjects, elevated RS-NOs values were observed in patients with CF (0.35 ± 0.07 µM, p < 0.01), with COPD (0.24 ± 0.04 µM, p < 0.01), and in smokers (0.46 ± 0.09 µM, p < 0.01).
In current smokers there was a correlation (r = 0.8, p < 0.05) between RS-NOs values and smoking history (pack/year).
Exhaled NO2
NO2 were detectable in all subjects (Figure 2). Compared
with normal values (0.45 ± 0.06 µM), exhaled NO2 levels
were higher in patients with severe asthma (1.04 ± 0.19 µM, p < 0.05), in those with CF (1.46 ± 0.3 µM, p < 0.01) and in those
with COPD (2.62 ± 0.52 µM, p < 0.01). No differences in exhaled NO2 values in patients with mild asthma (0.63 ±0.14 µM)
or in smokers (0.44 ±0.08 µM) were found when compared
with normal values.
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Exhaled NO
Exhaled NO levels are shown in Figure 3. Exhaled NO levels were significantly higher in patients with mild asthma (17.9 ± 1.8 ppb) than in control subjects (6.8 ± 0.3 ppb, p < 0.01).
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Exhaled NO was higher in patients with severe asthma (12.01 ± 2.1 ppb) than in control subjects (p < 0.05), but lower than levels in subjects with mild asthma (p < 0.05). No significant differences in exhaled NO values between patients with CF and control subjects. Exhaled NO was higher in those with COPD (12.01 ± 1.3 ppb) than in control subjects (p < 0.01). Exhaled NO in smokers was lower (5.8 ± 0.3 ppb) when compared with control subjects (p < 0.05).
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DISCUSSION |
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TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
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In the present study we found that RS-NOs are detectable in EBC of humans and that RS-NOs values are increased in different inflammatory airway diseases. Our results show high RS-NOs levels in EBC of patients with severe asthma, COPD, or CF, and in smokers, but not in patients with mild asthma.
EBC is a noninvasive method to collect airway secretions. The measurement of RS-NOs in an easily accessible fluid such as EBC may have applications in the search for the role of nitrosative stress/antioxidant defense imbalance in the pathogenesis of inflammatory lung diseases may have clinical relevance as a noninvasive biomarker of nitrosative stress.
In patients with asthma, the higher RS-NOs values in severe compared with mild asthma suggest that the measurement of RS-NOs in EBC may be helping to classify asthma severity. We speculate that in severe asthma nitrosative stress is greater than in mild asthma and consequently, there is a stronger adaptive response via the oxidation of NO. This supports other evidence for increased oxidant stress in patients with severe asthma (19). In addition, because RS-NOs depends on the interaction between NO and its metabolites with GSH, it must be taken into account that GSH level in bronchial fluid is increased in patients with asthma (23). Furthermore, seven of eight patients with severe persistent asthma had received chronic oral steroid treatment, and corticosteroids have been demonstrated to increase GSH synthase in both animal models and in vitro systems (24, 25). Finally, airway acidification may be an additional stimulus leading to RS-NOs formation (5).
Our findings in patients with asthma are not in line with the report of Gaston and colleagues (26), which showed a RS-NOs deficiency in asthmatic children with respiratory failure. Several points may explain this discrepancy such as different biologic samples studied (tracheal specimens versus EBC), different techniques for measuring RS-NOs (chemiluminescence versus colorimetric) and different age of patients. Moreover, the asthmatic children studied by Gaston and colleagues were not chronically treated with steroids as ours were, and corticosteroids increase GSH production (24, 25).
As far as NO2 values in EBC, we did not find increased
levels of NO2 in EBC of patients with mild asthma, and this
observation accords with a recent report by Nightingale and
colleagues (27). On the contrary, in patients with severe
asthma NO2 levels were higher than those observed in the
control group, but no differences between NO2 values in
mild and in severe asthma were observed. We therefore suggest that RS-NOs may be a better index of NO biologic pathway than NO2 in studying asthma severity.
Exhaled NO values in mild and severe asthma measured in our patients confirm previous observation of elevated levels in asthma (4, 28). This is in agreement with the current view of exhaled NO as a useful new lung function test in asthma (6).
In normal smokers, we also found high RS-NOs values in EBC compared with low exhaled NO levels. The reason for low NO values in exhaled air of smokers is still debated (29). It is possible that high NO levels in cigarette smoke may induce the production of GSH, which reacts with NO to form GS-NO. In this regard, we found a positive correlation between smoking history (pack/year) and RS-NOs levels suggesting that nitrosative stress induced by smoke stimulates RS-NOs production. GSH may protect cells against smoke-induced injury (30).
The normal values of NO2 in smokers were unexpected.
Nevertheless, this may be related to the fact that NO2 is rapidly oxidized into stable bioactive oxidation end-products of
NO pathway such as NO3 and RS-NOs. The chemistry of NO
suggests that NO2 is a weak base with a short lifetime in
aqueous solution because of its rapid oxidation to NO3 and
RS-NOs (7). In smokers, the high concentration of free radicals inhaled by cigarette smoking may force the transformation of NO2 into RS-NOs.
In patients with CF, our data confirm the previous observation that exhaled NO2 is raised in contrast to exhaled NO
(31). This could be related to the fact that exhaled NO is rapidly degraded by oxidation to NOx. We further support this
hypothesis by observing high levels of RS-NOs in EBC of patients with CF. These findings suggest that NOx may be more
valuable than exhaled NO in assessing airway inflammation in
patients with CF. The increased levels of RS-NOs and NO2
we found in patients with CF seem to contradict the findings of Kelley and coworkers (32), which showed that iNOS is essentially absent in the epithelium of CF airways. A possible
explanation is that activated cNOS, rather than iNOS, are responsible for NO production in epithelial cells, or that the production of NOx is mainly derived from inflammatory cells infiltrating airway mucosa of these patients (31).
In patients with COPD, RS-NOs and NO2 values were
higher than in control subjects. This may be related to an enhanced nitrosative stress. In fact, in airways of patients with
COPD there is an increased number of neutrophils, which
produce numerous free radicals (33), causing the oxidation of
part of NO gas into NOx such as RS-NOs and NO2. This fits
nicely with the demonstration of high levels of NOx in sputum
of patients with clinically stable COPD (34). In patients with
COPD, exhaled NO values tended to be higher than in control subjects, but less than in patients with steroid-naive asthma. For this reason, though exhaled NO seems to be a good
marker of airway inflammation in asthma management (6),
the clinical utility of exhaled NO in COPD is still debated (35).
We are aware that in our study three groups of subjects (controls versus severe asthma and versus COPD) were not age-matched. This makes it difficult to compare biologic parameters, in particular oxidative stress related parameters since oxidative burden in the lung increases with age (36). Nevertheless, we in EBC and other investigators in BAL (13) did not find any correlation between RS-NOs levels and age.
In conclusion, our study has shown that RS-NOs are detectable in EBC of healthy subjects and of patients with various inflammatory airway diseases. Whether RS-NOs measurement represents merely an adaptive response to nitrosative stress into airways, or whether RS-NOs may play a more active role in protecting the airways against nitrosative stress, needs to be further studied.
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Footnotes |
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Correspondence and requests for reprints should be addressed to Prof. Peter J. Barnes, Department of Thoracic Medicine, National Heart & Lung Institute, Dovehouse Street, London SW3 6LY, UK. E-mail: p.j.barnes{at}ic.ac.uk
(Received in original form January 20, 2000 and in revised form January 9, 2001).
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References |
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ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
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M. Cazzola, W. MacNee, F. J. Martinez, K. F. Rabe, L. G. Franciosi, P. J. Barnes, V. Brusasco, P. S. Burge, P. M. A. Calverley, B. R. Celli, et al. Outcomes for COPD pharmacological trials: from lung function to biomarkers Eur. Respir. J., February 1, 2008; 31(2): 416 - 469. [Abstract] [Full Text] [PDF]
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 P. Montuschi Review: Analysis of exhaled breath condensate in respiratory medicine: methodological aspects and potential clinical applications Therapeutic Advances in Respiratory Disease, October 1, 2007; 1(1): 5 - 23. [Abstract] [PDF]
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 Y. Y. Tyurina, L. V. Basova, N. V. Konduru, V. A. Tyurin, A. I. Potapovich, P. Cai, H. Bayir, D. Stoyanovsky, B. R. Pitt, A. A. Shvedova, et al. Nitrosative Stress Inhibits the Aminophospholipid Translocase Resulting in Phosphatidylserine Externalization and Macrophage Engulfment: IMPLICATIONS FOR THE RESOLUTION OF INFLAMMATION J. Biol. Chem., March 16, 2007; 282(11): 8498 - 8509. [Abstract] [Full Text] [PDF]
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 P. J. Barnes, B. Chowdhury, S. A. Kharitonov, H. Magnussen, C. P. Page, D. Postma, and M. Saetta Pulmonary Biomarkers in Chronic Obstructive Pulmonary Disease Am. J. Respir. Crit. Care Med., July 1, 2006; 174(1): 6 - 14. [Abstract] [Full Text] [PDF]
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H. Kanazawa and J. Yoshikawa Elevated Oxidative Stress and Reciprocal Reduction of Vascular Endothelial Growth Factor Levels With Severity of COPD Chest, November 1, 2005; 128(5): 3191 - 3197. [Abstract] [Full Text] [PDF]
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I. Horvath, J. Hunt, P. J. Barnes, and On behalf of the ATS/ERS Task Force on Exhaled Bre Exhaled breath condensate: methodological recommendations and unresolved questions Eur. Respir. J., September 1, 2005; 26(3): 523 - 548. [Abstract] [Full Text] [PDF]
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 W. MacNee Treatment of stable COPD: antioxidants Eur. Respir. Rev., September 1, 2005; 14(94): 12 - 22. [Abstract] [Full Text] [PDF]
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 W. MacNee Pulmonary and Systemic Oxidant/Antioxidant Imbalance in Chronic Obstructive Pulmonary Disease Proceedings of the ATS, April 1, 2005; 2(1): 50 - 60. [Abstract] [Full Text] [PDF]
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R. Buhl and S. G. Farmer Future Directions in the Pharmacologic Therapy of Chronic Obstructive Pulmonary Disease Proceedings of the ATS, April 1, 2005; 2(1): 83 - 93. [Abstract] [Full Text] [PDF]
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 H Marteus, D C Tornberg, E Weitzberg, U Schedin, and K Alving Origin of nitrite and nitrate in nasal and exhaled breath condensate and relation to nitric oxide formation Thorax, March 1, 2005; 60(3): 219 - 225. [Abstract] [Full Text] [PDF]
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 P. J. Barnes Mediators of Chronic Obstructive Pulmonary Disease Pharmacol. Rev., December 1, 2004; 56(4): 515 - 548. [Abstract] [Full Text] [PDF]
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S. A. Kharitonov and P. J. Barnes Effects of Corticosteroids on Noninvasive Biomarkers of Inflammation in Asthma and Chronic Obstructive Pulmonary Disease Proceedings of the ATS, November 1, 2004; 1(3): 191 - 199. [Abstract] [Full Text] [PDF]
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K. M. Beeh, J. Beier, N. Koppenhoefer, and R. Buhl Increased Glutathione Disulfide and Nitrosothiols in Sputum Supernatant of Patients With Stable COPD Chest, October 1, 2004; 126(4): 1116 - 1122. [Abstract] [Full Text] [PDF]
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C. R. Morris, M. Poljakovic, L. Lavrisha, L. Machado, F. A. Kuypers, and S. M. Morris Jr. Decreased Arginine Bioavailability and Increased Serum Arginase Activity in Asthma Am. J. Respir. Crit. Care Med., July 15, 2004; 170(2): 148 - 153. [Abstract] [Full Text] [PDF]
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 N. L. Reynaert, K. Ckless, S. H. Korn, N. Vos, A. S. Guala, E. F. M. Wouters, A. van der Vliet, and Y. M. W. Janssen-Heininger From the Cover: Nitric oxide represses inhibitory {kappa}B kinase through S-nitrosylation PNAS, June 15, 2004; 101(24): 8945 - 8950. [Abstract] [Full Text] [PDF]
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G. E. Carpagnano, P. J. Barnes, J. Francis, N. Wilson, A. Bush, and S. A. Kharitonov Breath Condensate pH in Children With Cystic Fibrosis and Asthma: A New Noninvasive Marker of Airway Inflammation? Chest, June 1, 2004; 125(6): 2005 - 2010. [Abstract] [Full Text] [PDF]
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P.G.A. Van Hoydonck, W.A. Wuyts, B.M. Vanaudenaerde, E.G. Schouten, L.J. Dupont, and E.H.M. Temme Quantitative analysis of 8-isoprostane and hydrogen peroxide in exhaled breath condensate Eur. Respir. J., February 1, 2004; 23(2): 189 - 192. [Abstract] [Full Text] [PDF]
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 H.-W. Shin, C. M. Rose-Gottron, D. M. Cooper, R. L. Newcomb, and S. C. George Airway diffusing capacity of nitric oxide and steroid therapy in asthma J Appl Physiol, January 1, 2004; 96(1): 65 - 75. [Abstract] [Full Text] [PDF]
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A.W. Boots, G.R.M.M. Haenen, and A. Bast Oxidant metabolism in chronic obstructive pulmonary disease Eur. Respir. J., November 2, 2003; 22(46_suppl): 14s - 27s. [Abstract] [Full Text] [PDF]
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E. Baraldi, L. Ghiro, V. Piovan, S. Carraro, G. Ciabattoni, P. J. Barnes, and P. Montuschi Increased Exhaled 8-Isoprostane in Childhood Asthma Chest, July 1, 2003; 124(1): 25 - 31. [Abstract] [Full Text] [PDF]
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V. L. Kinnula and J. D. Crapo Superoxide Dismutases in the Lung and Human Lung Diseases Am. J. Respir. Crit. Care Med., June 15, 2003; 167(12): 1600 - 1619. [Abstract] [Full Text] [PDF]
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M. Corradi, I. Rubinstein, R. Andreoli, P. Manini, A. Caglieri, D. Poli, R. Alinovi, and A. Mutti Aldehydes in Exhaled Breath Condensate of Patients with Chronic Obstructive Pulmonary Disease Am. J. Respir. Crit. Care Med., May 15, 2003; 167(10): 1380 - 1386. [Abstract] [Full Text] [PDF]
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H Kanazawa, S Shiraishi, K Hirata, and J Yoshikawa Imbalance between levels of nitrogen oxides and peroxynitrite inhibitory activity in chronic obstructive pulmonary disease Thorax, February 1, 2003; 58(2): 106 - 109. [Abstract] [Full Text] [PDF]
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S A Kharitonov, L E Donnelly, P Montuschi, M Corradi, J V Collins, and P J Barnes Dose-dependent onset and cessation of action of inhaled budesonide on exhaled nitric oxide and symptoms in mild asthma Thorax, October 1, 2002; 57(10): 889 - 896. [Abstract] [Full Text] [PDF]
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E. Demoncheaux, D. Crowther, A. C. Spivey, T. W. Higenbottam, P. J. Barnes, and S. A. Kharitonov Monitoring reactive nitrogen species in biological milieu: A difficult journey Am. J. Respir. Crit. Care Med., June 15, 2002; 165(12): 1670 - 1671. [Full Text] [PDF]
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W. Formanek, D. Inci, R.P. Lauener, J.H. Wildhaber, U. Frey, and G.L. Hall Elevated nitrite in breath condensates of children with respiratory disease Eur. Respir. J., March 1, 2002; 19(3): 487 - 491. [Abstract] [Full Text] [PDF]
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M. J. TOBIN Asthma, Airway Biology, and Nasal Disorders in AJRCCM 2001 Am. J. Respir. Crit. Care Med., March 1, 2002; 165(5): 598 - 618. [Full Text] [PDF]
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R. M. EFFROS, K. W. HOAGLAND, M. BOSBOUS, D. CASTILLO, B. FOSS, M. DUNNING, M. GARE, W. LIN, and F. SUN Dilution of Respiratory Solutes in Exhaled Condensates Am. J. Respir. Crit. Care Med., March 1, 2002; 165(5): 663 - 669. [Abstract] [Full Text] [PDF]
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H.-W. SHIN, C. M. ROSE-GOTTRON, R. S. SUFI, F. PEREZ, D. M. COOPER, A. F. WILSON, and S. C. GEORGE Flow-independent Nitric Oxide Exchange Parameters in Cystic Fibrosis Am. J. Respir. Crit. Care Med., February 1, 2002; 165(3): 349 - 357. [Abstract] [Full Text] [PDF]
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G. L. Hall, B. Reinmann, J. H. Wildhaber, and U. Frey Tidal exhaled nitric oxide in healthy, unsedated newborn infants with prenatal tobacco exposure J Appl Physiol, January 1, 2002; 92(1): 59 - 66. [Abstract] [Full Text] [PDF]
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O. W. Griffith Glutaminase and the Control of Airway pH . Yet Another Problem for the Asthmatic Lung? Am. J. Respir. Crit. Care Med., January 1, 2002; 165(1): 1 - 2. [Full Text] [PDF]
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G. M. MUTLU, K. W. GAREY, R. A. ROBBINS, L. H. DANZIGER, and I. RUBINSTEIN Collection and Analysis of Exhaled Breath Condensate in Humans Am. J. Respir. Crit. Care Med., September 1, 2001; 164(5): 731 - 737. [Full Text] [PDF]
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