|
|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
 |
ABSTRACT |
|---|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
The reaction of nitric oxide (NO) and superoxide anions (O2
) in
the airway results in the formation of peroxynitrite, a highly reactive oxidant species. Peroxynitrite reacts with tyrosine residues in
proteins to form the stable product nitrotyrosine. We investigated whether nitrotyrosine in exhaled breath condensates may be increased in patients with asthma. Four groups of nonsmoking subjects were studied. We measured exhaled NO, nitrotyrosine, and
leukotrienes concentrations in breath condensate in healthy nonatopic subjects (n = 15) and in patients with mild asthma (steroid
naive, n = 15), moderate asthma (inhaled steroid treatment, n = 12), and severe asthma (oral steroid treatment, n = 12). Exhaled
NO was increased significantly in patients with mild (19.2 ± 2.7 ppb, p < 0.01) and moderate asthma (14.0 ± 1.53 ppb, p < 0.05),
as compared with normal control (6.58 ± 0.61 ppb). The levels of LTC4/D4/E4 and LTB4 were increased significantly in patients with moderate and severe asthma treated with steroids. Nitrotyrosine concentrations were detectable (6.3 ± 0.8 ng/ml) in breath condensate of normal subjects, and were increased significantly in patients with mild asthma (15.3 ± 2.0 ng/ml, p < 0.01). However,
the levels of nitrotyrosine in exhaled condensate were lower in patients with moderate (5.0 ± 0.6 ng/ml) and severe asthma (3.3 ± 0.6 ng/ml, p < 0.05). There was a significant correlation between
nitrotyrosine in breath condensate and exhaled NO in patients
with mild asthma (r = 0.65, p < 0.05). We conclude that nitrotyrosine formation in exhaled breath condensates may be a marker
of oxidative stress in airways of asthma.
| |
INTRODUCTION |
|---|
|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
Oxidative stress, defined as an increased exposure to oxidants
and/or decreased antioxidant capacities, is implicated in airway inflammatory airway diseases, such as chronic obstructive
pulmonary disease (COPD) and asthma (1). Inflammatory
cells, especially eosinophils, which produce more superoxide
anions (O2) than neutrophils or macrophages, release several
reactive oxygen- and nitrogen-derived species such as nitric
oxide (NO). Levels of NO are elevated in the air exhaled by
patients with asthma (4), and may contribute to airway edema
and inflammation (5). High levels of O2 have also been detected in bronchoalveolar lavage of patients with asthma, and
the concentration was inversely correlated with forced expiratory volume in 1 s (FEV1) (6). A reaction between NO and
O2 results in the formation of peroxynitrite anions, a highly
reactive oxidant species (7). Peroxynitrite adds a nitro group
to the 3-position adjacent to the hydroxyl group of tyrosine to
produce the stable product nitrotyrosine (8). Peroxynitrite induces hyperresponsiveness in airways of guinea pigs (9), inhibits pulmonary surfactant (10), and damages pulmonary epithelial cells (11). The formation of peroxynitrite is now well
established in various forms of airway diseases in the lung, including endotoxic shock, adult respiratory distress syndrome
(ARDS), hyperoxia, and ischemia-reperfusion injury (7, 12,
13). Recent study also suggests that there was strong immunoreactivity for nitrotyrosine in the airway epithelium, lung
parenchyma, and inflammatory cells in the airways of patients
with asthma (14, 15), and that inhaled steroid treatment resulted in a significant reduction in nitrotyrosine immunoreactivity (14). Furthermore, a number of in vitro studies have established changes in enzyme activity on nitration of critical
tyrosine residues, which has raised suggestions that protein nitration in vivo may be causally linked to inflammation-related forms of lung injury (16, 17).
The cysteinyl-leukotrienes (Cys-LTs: LTC4/D4/E4) are released from chopped human lung from allergic subjects in response to challenge with allergen (18). Calhoun and coworkers reported that zafirlukast, a LTD4 receptor antagonist, significantly blunted the antigen-derived augmentation of superoxide release by alveolar macrophages after challenge in patients with asthma (19). This study provides evidence that Cys-LTs can also modify cell activation in allergic inflammation, and generate reactive oxygen species. By contrast, LTB4 is a potent chemoattractant and activator of neutrophils, without significant effects on airway smooth muscle (20). LTB4 also activates the NADPH oxidase in guinea pig eosinophils, causing hydrogen peroxide (H2O2) generation and enhancing inflammation by oxidative stress (21). These findings suggest that Cys-LTs and LTB4 may be important mediators in the pathogenesis of asthma, including bronchial smooth muscle contraction, mucus production, inflammatory cell recruitment, and enhancement of oxidative stress.
The aim of this study was to examine whether nitrotyrosine could be detected in breath condensate of patients with asthma, as a marker of oxidative stress in airways of patients with asthma, and to investigate the effect of steroid treatment on nitrotyrosine production in breath condensates of patients with asthma, comparing LT release by inflammatory cells.
| |
METHODS |
|---|
|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
Patients
Four groups of nonsmoking subjects were studied. Fifteen healthy nonatopic subjects and 15 patients with mild asthma, 12 with moderate
asthma, and 12 with severe asthma were recruited (Table 1). There
was no significant difference in ages among the subject groups. Atopy
was assessed by skin prick tests for common allergens. The diagnosis
of bronchial asthma was based on the criteria of the American Thoracic Society (22). Severity of asthma was classified according to the
National Institutes of Health/World Health Organization (NIH/
WHO) guidelines (23). Briefly, subjects with mild asthma had symptoms twice a week or less often, with an FEV1 
80% predicted, and
were taking regular medication, but used inhaled 
2-agonists as needed
for symptom relief. Subjects with moderate asthma had daily symptoms, used inhaled short-acting 2-agonists daily, had an FEV1 between 60% and 80% predicted, and were taking regular inhaled glucocorticoids (budesonide, 0.4 to 3.2 mg; fluticasone propionate, 0.5 to
2 mg; or beclomethasone, 1 to 2 mg). Subjects with severe asthma were
treated with oral prednisolone (4 to 50 mg/d) and inhaled steroids (fluticasone propionate: 1 to 4 mg/d, or budesonide: 0.8 to 4 mg/d). The
protocol was approved by the Ethics Committee of the Royal Brompton Hospital, and informed consent from each subject was obtained.
|
Study Design
Subjects' details were obtained and then baseline spirometry (Vitalograph Ltd., Buckingham, UK) and exhaled NO were measured, followed by collection of expired breath condensate.
Expired Breath Condensate
Expired breath condensate was collected by using a condenser, which
allowed the noninvasive collection of nongaseous components of the
expiratory air (EcoScreen, Jaeger, Würzburg, Germany). Subjects
breathed through a mouthpiece and a two-way nonrebreathing valve,
which also served as a saliva trap. They were asked to breathe at a
normal frequency and tidal volume, wearing a noseclip, for a period
of 10 min. The condensate, at least 1 ml, was collected as ice at 
20° C
and stored at 70° C immediately.
Nitrotyrosine and Leukotrienes Measurements
Nitrotyrosine was measured with a specific enzyme immunoassay (EIA) (Cayman Chemical, Ann Arbor, MI). Initially, assays were performed on unconcentrated condensate samples. The lower limit of detection for this assay was 3.9 ng/ml. If nitrotyrosine was not detected in unconcentrated condensate samples, the breath condensates were concentrated threefold, using a freeze dryer (Modulyo; Edwards, Crawley, UK), and reanalyzed. If nitrotyrosine was not detected even after threefold concentration, a value of 1.3 ng/ml (equal to the lower limit of detection, 3.9 ng/ml, divided by 3 to account for the threefold concentration of breath condensates) was arbitrarily assigned to it. LTC4/D4/E4 and LTB4 concentrations were also measured by EIA (LTC4/D4/E4; Amersham Pharmacia Biotech, Amersham, UK; LTB4, Cayman Chemical). The lower limits of detection for these assays were 15.0 pg/ml and 4.4 pg/ml, respectively. If LTs were not detected in the breath condensate samples, a value of 15.0 pg/ml for LTC4/D4/E4 or 4.4 pg/ml for LTB4 was arbitrarily assigned to it.
Exhaled NO Measurement
Exhaled NO was measured by a chemiluminescence analyzer (Model LR2000; Logan Research, Rochester, UK), sensitive to NO from 1 to 500 ppb by volume, and with a resolution of 0.3 ppb. The analyzer was designed for online recording of exhaled NO concentrations. It was calibrated with certified NO mixtures (90 and 436 ppb) in nitrogen (BOC Special Gases, Guilford, UK). Measurements of exhaled NO were made by slow exhalation (5 to 6 L/min) from TLC for 20 to 30 s against a resistance (3 ± 0.4 mm Hg), to prevent nasal contamination.
Statistical Analysis
One-way analysis of variance (ANOVA) with the Newman-Keuls test for multiple comparisons was used to compare groups. Linear regression analysis was used to assess the relationship between nitrotyrosine concentrations in breath condensate and exhaled NO. Data were expressed as means ± SEM, and significance was defined as a value of p < 0.05.
| |
RESULTS |
|---|
|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
Nitrotyrosine concentrations were detectable (6.3 ± 0.8 ng/ml) in breath condensate of normal subjects, and were increased significantly in patients with mild asthma (15.3 ± 2.0 ng/ml, p < 0.01), who were not treated with steroids, as compared with normal control (Figure 1). However, the levels of nitrotyrosine in exhaled condensate were lower in patients with moderate (5.0 ± 0.6 ng/ml) and severe asthma (3.3 ± 0.6 ng/ml, p < 0.05), who were treated with inhaled or oral corticosteroids, than those of control subjects (Figure 1). There was no correlation between nitrotyrosine level and age in subjects in any group. Exhaled NO was increased significantly in patients with mild (19.2 ± 2.7 ppb, p < 0.01) and moderate asthma (14.0 ± 1.53 ppb, p < 0.05), as compared with normal control (6.58 ± 0.61 ppb) (Figure 1). But there was no significant difference in exhaled NO between normal control subjects and patients with severe asthma with steroid treatment. In patients with mild asthma, the levels of nitrotyrosine in exhaled breath condensate correlated with exhaled NO (r = 0.65, p < 0.05) (Figure 2). No correlation was found between nitrotyrosine concentrations in breath condensate and FEV1 in patients with mild asthma (data not shown). The levels of LTC4/D4/E4 and LTB4 in breath condensate were elevated significantly in patients with moderate (19.5 ± 1.3 and 161.0 ± 31.2 pg/ml, respectively, p < 0.01) and severe asthma (21.2 ± 1.7 and 186.8 ± 31.2 pg/ml, p < 0.01), compared with normal subjects (15.5 ± 0.2 and 63.1 ± 17.3 pg/ml) and patients with mild asthma (15.8 ± 0.3 and 79.1 ± 20.9 pg/ml) (Figure 3). There was no correlation between LTC4/D4/E4 or LTB4 and FEV1 in patients with moderate and severe asthma, however (data not shown).
|
|
|
| |
DISCUSSION |
|---|
|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
We demonstrated that exhaled nitrotyrosine was increased in
patients with mild asthma who were not treated with corticosteroids. Saleh and coworkers reported strong immunoreactivity for nitrotyrosine in the airway epithelium and inflammatory cells in bronchial biopsies of patients with asthma, which
was not seen in normal control subjects, and was reduced by
inhaled corticosteroid treatment (14). A recent study also demonstrated intense nitrotyrosine immunoreactivity was in the
airways and lung parenchyma of patients with asthma who
died of status asthmaticus despite steroid treatment, and suggested that status asthmaticus is characterized by a failure of
corticosteroids to control the formation of reactive nitrogen species (15). In addition, nitration of proteins in bronchoalveolar lavage fluid was increased in patients with acute respiratory distress syndrome receiving inhaled nitric oxide (24). This evidence suggests that high levels of NO and O2 produced in
the airways may react to form the potent oxidant, peroxynitrite (12). Peroxynitrite causes hyperresponsiveness in airways
of guinea pigs (9) and respiratory epithelial damage (11). Peroxynitrite-induced protein nitration may also alter protein
function; peroxynitrite inactivates surfactant (25) and inhibits
protein phosphorylation by tyrosine kinases, thus interfering
with signal transduction mechanisms (26). Furthermore, it has
been suggested that eosinophil peroxidase (EPO) and myeloperoxidase (MPO)-catalyzed nitration in the presence of
hydrogen peroxide (H2O2) to form nitrating intermediates
from nitrite (NO2), a main end product of NO, as an alternative mechanism of protein nitration, independent of peroxynitrite (27, 28). Recent studies suggest that tyrosine nitration by
peroxynitrite on eosinophil chemoattractant such as RANTES
and interleukin-5 (IL-5) may be a mechanism altering their
binding and chemoatactic function (29). These findings have
supported the hypothesis that tyrosine nitration might result
not only in the formation of inactive "footprints" of reactive
nitrogen intermediates, but might also be functionally related
to the pathobiology of airway inflammatory diseases. In this
study we have provided strong evidence that oxidative stress induced by inflammation produces nitrotyrosine, which presumably reflects increased peroxynitrite formation or direct
nitration by granulocyte peroxidases. The increased nitrated
proteins are presumably found in the airway lumen and collected in the expired condensate of patients with mild asthma
who are not treated with corticosteroids.
In our study there was a positive correlation between exhaled NO and nitrotyrosine level in breath condensate. There
is now persuasive evidence that levels of NO are increased in
association with airway inflammation and are decreased by
antiinflammatory treatments (30). Exhaled NO has a positive
correlation with eosinophil counts in induced sputum in patients with asthma (31). We therefore investigated whether nitrotyrosine concentrations were related with any other markers of airway inflammation, which was detectable in breath
condensates and contributed to oxidative stress in airways of
asthma, and found significant increases in Cys-LTs and LTB4
in breath condensates of patients with moderate and severe
asthma, compared with normal subjects and subjects with mild
asthma. The Cys-LTs, generated predominantly by mast cells
and eosinophils, possess various bronchoactive properties in
vitro, inducing airway smooth muscle contraction (32, 33), microvascular leakage, and mucus hypersecretion (32). A recent study has provided evidence that zafirlukast, the potent Cys-LT receptor antagonist, reduced the number of eosinophils
and basophils recovered in bronchoalveolar lavage fluid in patients with asthma after allergen challenge, and blunted the
antigen-derived augmentation of O2 release by alveolar macrophages after challenge in patients with asthma (19). In addition, LTB4 is a potent proinflammatory mediator and an attractant for neutrophils. Although LTB4 has not been closely
linked to asthma in contrast to the Cys-LTs, there is increasing
evidence that neutrophils may play a role in more severe asthma. Neutrophil number and activation are increased in
the airways of subjects with severe persistent asthma and during exacerbations of asthma (34, 35). In our study the levels of
LTB4 in breath condensate were significantly higher in patients with moderate and severe asthma than those of normal
subjects and patients with mild asthma. This suggests that
LTB4 may be involved in exacerbations of asthma, probably in
neutrophil recruitment. LTB4 also directly promotes a receptor-mediated activation of the NADPH oxidase in guinea pig
eosinophils, to cause the formation of H2O2 (21). LTB4 presumably induces neutrophil recruitment and activation of
MPO, resulting in nitrotyrosine production in airways of patients with asthma. However, there was no significant increase
in either exhaled NO or nitrotyrosine in breath condensates of
patients with moderate and severe asthma whose levels of
Cys-LTs and LTB4 in condensates were increased significantly. This finding suggests that an increase in NO production plays a critical role in nitrotyrosine production in airways
of patients with asthma.
We found a significant reduction in nitrotyrosine production in breath condensates of patients with severe asthma who required systemic treatment with corticosteroids, compared with healthy control subjects. In contrast, there was no significant difference in exhaled NO between normal control subjects and subjects with severe asthma. This finding suggests that systemic steroid treatment may inhibit oxidative stress induced by inflammation and also inhibits the inflammatory response in the airways. The finding that levels of exhaled nitrotyrosine were lower than in normal subjects suggests that there may be some inflammation and oxidative stress in normal airways as a result of inhalation of oxidants in the urban environment. The recognition of asthma as a chronic inflammatory disease (3), along with the recommendation by asthma management guidelines (23) for early intervention with antiinflammatory agents, has resulted in inhaled corticosteroid therapy becoming the mainstay of steroid treatment for patients with asthma. Despite the availability and use of inhaled corticosteroids for the treatment of asthma, a proportion of patients require long-term adjunct therapy with systemic corticosteroids to achieve control of symptoms. Because of the recognized risk of side effects associated with the use of systemic corticosteroids, the development of therapeutic options that can eliminate the dependence of these patients on oral corticosteroids is important. Our data suggest that expired nitrotyrosine may be a useful marker to control the dose of systemic corticosteroids, or to combine new antiasthma drugs with inhaled steroids, keeping nitrotyrosine in breath condensate at normal levels.
In summary, our study has demonstrated that nitrotyrosine in exhaled breath condensate was increased in patients with mild asthma who were not treated with corticosteroids, compared with normal control subjects, and was reduced in patients with severe asthma receiving steroid therapy. We have also provided evidence that nitrotyrosine formation in exhaled breath condensate may be a more sensitive marker to evaluate the contribution of oxidative stress to airway inflammation of asthma than exhaled NO.
| |
Footnotes |
|---|
Correspondence and requests for reprints should be addressed to Peter J. Barnes, D.M., Department of Thoracic Medicine, Imperial College School of Medicine at the National Heart and Lung Institute, Dovehouse Street, London SW3 6LY, UK. E-mail: p.j.barnes{at}ic.ac.uk
(Received in original form December 14, 1999 and in revised form March 17, 2000).
| |
References |
|---|
|
TOP
ABSTRACT
INTRODUCTION
METHODS
RESULTS
DISCUSSION
REFERENCES
|
|---|
1. Rahman, I., D. Morrison, K. Donaldson, and W. MacNee. 1996. Systemic oxidative stress in asthma, COPD and smokers. Am. J. Respir. Crit. Care Med. 154: 1055-1060 [Abstract].
2.
Repine, J. E.,
A. Bast,
I. Lankhorst, and
the Oxidative Stress Study
Group.
1997.
Oxidative stress in chronic obstructive pulmonary disease. State of art.
Am. J. Respir. Crit. Care Med.
156:
341-357
3. Djukanovic, F., W. R. Roche, J. W. Wilson, C. R. W. Beasley, O. P. Twentyman, P. H. Howarth, and S. T. Holgate. 1990. Mucosal inflammation in asthma. Am. Rev. Respir. Dis. 142: 434-457 [Medline].
4.
Barnes, P. J., and
S. A. Kharitonov.
1996.
Exhaled nitric oxide
a new
lung function test.
Thorax
51:
233-237
5. Barnes, P. J.. 1996. Pathophysiology of asthma. Br. J. Clin. Pharmacol. 42: 3-10 [Medline].
6. Jarjour, N. N., and W. J. Calhoun. 1994. Enhanced production of oxygen radicals in asthma. J. Lab. Clin. Med. 123: 131-136 [Medline].
7.
Beckman, J. S.,
T. W. Beckman,
J. Chen,
P. A. Marshall, and
B. A. Freeman.
1990.
Apparent hydroxyl radical production by peroxynitrite:
implication for endothelial injury from nitric oxide and superoxide.
Proc. Natl. Acad. Sci. U.S.A.
87:
1620-1624
8. Ischiropoulos, H., L. Zhu, J. Chen, M. Tsai, J. C. Martin, C. D. Smith, and J. S. Beckman. 1992. Peroxynitrite-mediated tyrosine nitration catalyzed by superoxide dismutase. Arch. Biochem. Biophys. 298: 438-445 [Medline].
9. Sadeghi-Hashjin, G., G. Folkerts, P. A. Henricks, A. K. C. P. Verheyen, H. J. Linde, I. Ark, A. Coene, and F. P. Nijkamp. 1996. Peroxynitrite induces airway hyperresponsiveness in guinea pigs in vitro and in vivo. Am. J. Respir. Crit. Care Med. 153: 1697-1701 [Abstract].
10.
Haddad, I. Y.,
H. Ischiropoulos,
B. A. Holm,
J. S. Beckman,
J. R. Baker, and
S. Matalon.
1993.
Mechanisms of peroxynitrite induced injury to
pulmonary surfactants.
Am. J. Physiol.
265:
L555-L564
11. Matalon, S., P. Hu, H. Ischiropoulos, and J. S. Beckman. 1994. Peroxynitrite inhibition of oxygen consumption and ion transport in alveolar type II pneumocytes. Chest 105: 74S .
12. Wizemann, T., C. Gardner, J. Laskin, S. Quinones, S. Durham, N. Goller, T. Ohnishi, and D. Laskin. 1994. Production of nitric oxide and peroxynitrite in the lung during acute endotoxemia. J. Leukocyte Biol. 56: 759-768 [Abstract].
13.
Ischiropoulos, H.,
A. B. al-Mehdi, and
A. B. Fisher.
1995.
Reactive species in ischemic rat lung injury: contribution of peroxynitrite.
Am. J. Physiol.
269:
L158-L164
14.
Saleh, D.,
P. Ernst,
S. Lim,
P. J. Barnes, and
A. Giaid.
1998.
Increased
formation of the potent oxidant peroxynitrite in the airways of asthmatic patients is associated with induction of nitric oxide synthase: effect of inhaled glucocorticoid.
FASEB J.
12:
929-937
15. Kaminsky, D. A., J. Mitchell, N. Carroll, A. James, R. Soultanakis, and Y. Janssen. 1999. Nitrotyrosine formation in the airways and lung parenchyma of patients with asthma. J. Allergy Clin. Immunol. 104: 747-754 [Medline].
16. Yermilov, V., J. Rubio, and H. Ohshima. 1995. Formation of 8-nitroguanine in DNA treated with peroxynitrite in vitro and its rapid removal from DNA by depurination. FEBS Lett. 376: 207-210 [Medline].
17. Juedes, M. J., and G. N. Wogan. 1996. Peroxynitrite-induced mutation spectra of pSP189 following replication in bacteria and in human cells. Mutation Res. 349: 51-61 .
18.
Dahlen, S. E.,
G. Hasson,
P. Hedqvist,
T. Bjorck,
E. Gramstrom, and
B. Dahlen.
1983.
Allergen challenge of lung tissue from asthmatics elicits
bronchial contraction that correlates with the release of leukotrienes
C4, D4, and E4.
Proc. Natl. Acad. Sci. U.S.A.
80:
1712-1716
19.
Calhoun, W. J.,
B. J. Lavins,
M. C. Minkwitz,
R. Evans,
G. J. Gleich, and
J. Cohn.
1998.
Effect of zafirlukast (Accolate) on cellular mediators of
inflammation.
Am. J. Respir. Crit. Care Med.
157:
1381-1389
20. Claesson, H. E., B. Odlander, and P. J. Jakobssen. 1992. Leukotriene B4 in the immune system. Int. J. Immunopharmacol. 14: 441-449 [Medline].
21.
Lindsay, M. A.,
R. S. Perkins,
P. J. Barnes, and
M. A. Giembycz.
1998.
Leukotriene B4 activates the NADPH oxidase in eosinophils by a pertussis toxin-sensitive mechanism that is largely independent of arachidonic acid mobilization.
J. Immunol.
160:
4526-4534
22. American Thoracic Society. 1987. Standards for the diagnosis and care of patients with chronic obstructive pulmonary disease (COPD) and asthma. Am. Rev. Respir. Dis. 136: 225-244 [Medline].
23. National Institutes of Health: National Heart, Lung, and Blood Institute. 1997. Guidelines for the diagnosis and management of asthma. National Institutes of Health, Washington, DC. Publication No. 97-4051.
24.
Lamb, N. J.,
G. J. Quinlan,
S. T. Westerman,
J. M. C. Gutteridge, and
T. W. Evans.
1999.
Nitration of proteins in bronchoalveolar lavage
fluid from patients with acute respiratory distress syndrome receiving
inhaled nitric oxide.
Am. J. Respir. Crit. Care Med.
160:
1031-1034
25.
Haddad, I. Y.,
J. Crow,
Y. Yoazu,
J. S. Beckman, and
S. Matalon.
1994.
Concurrent generation of nitric oxide and superoxide damages surfactant protein A (SP-A).
Am. J. Physiol.
267:
L242-L249
26.
Grow, A. J.,
D. G. Buerk, and
H. Ischiropoulos.
1997.
A novel reaction
mechanism for the formation of S-nitrosothiol in vivo.
J. Biol. Chem.
272:
2841-2845
27.
Wu, W.,
Y. Chen, and
S. L. Hazen.
1999.
Eosinophil peroxidase nitrates
protein tyrosyl residues: implications for oxidative damage by nitrating intermediates in eosinophilic inflammatory disorders.
J. Biol.
Chem.
274:
25833-25944
28. Eiserich, J. P., M. Hristova, C. E. Cross, A. D. Jones, B. A. Freeman, B. Halliwell, and A. van der Vliet. 1998. Formation of nitric oxide-derived inflammatory oxidants by myeloperoxidase in neutrophils. Nature 391: 393-397 [Medline].
29.
Sato, E.,
K. L. Simpson,
M. B. Grisham,
S. Koyama, and
R. A. Robbins.
1999.
Effects of reactive oxygen and nitrogen metabolites on
RANTES- and IL-5-induced eosinophil chemoatactic activity in vitro.
Am. J. Pathol.
155:
591-598
30. Kharitonov, S. A., D. H. Yates, and P. J. Barnes. 1996. Inhaled glucocorticoids decrease nitric oxide in exhaled air of patients with asthma. Am. J. Respir. Crit. Care Med. 153: 454-457 [Abstract].
31. Jatakanon, A., S. Lim, S. A. Kharitonov, K. F. Chung, and P. J. Barnes. 1998. Correlation between exhaled nitric oxide, sputum eosinophils, and methacholine responsiveness in patients with mild asthma. Thorax 53: 91-95 [Abstract].
32. Lewis, R. A., K. F. Austen, and R. J. Soberman. 1990. Leukotrienes and other products of the 5-lipoxygenase pathway: biochemistry and relation to pathobiology in human disease. N. Engl. J. Med. 323: 645-655 [Medline].
33. Rabe, K. F., N. M. Munoz, A. J. Vita, B. E. Morton, H. Magnussen, and A. R. Leff. 1994. Contraction of human bronchial smooth muscle caused by activated human eosinophils. Am. J. Physiol. 267: 1326-1334 .
34.
Jatakanon, A.,
C. Uasuf,
W. Maziak,
S. Lim,
K. F. Chung, and
P. J. Barnes.
1999.
Neutrophilic inflammation in severe persistent asthma.
Am.
J. Respir. Crit. Care Med.
160:
1532-1539
35. Fahy, J. V., K. W. Kim, J. Liu, and H. A. Boushey. 1995. Prominent neutrophilic inflammation in sputum from subjects with asthma exacerbation. J. Allergy Clin. Immunol. 95: 843-852 [Medline].
This article has been cited by other articles:
 |
|
 |
|
  H. Maarsingh, B. E. Bossenga, I. S. T. Bos, H. H. Volders, J. Zaagsma, and H. Meurs L-Arginine deficiency causes airway hyperresponsiveness after the late asthmatic reaction Eur. Respir. J., July 1, 2009; 34(1): 191 - 199. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. O. Osoata, T. Hanazawa, C. Brindicci, M. Ito, P. J. Barnes, S. Kharitonov, and K. Ito Peroxynitrite Elevation in Exhaled Breath Condensate of COPD and Its Inhibition by Fudosteine Chest, June 1, 2009; 135(6): 1513 - 1520. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Brindicci, K. Ito, O. Torre, P. J. Barnes, and S. A. Kharitonov Effects of Aminoguanidine, an Inhibitor of Inducible Nitric Oxide Synthase, on Nitric Oxide Production and Its Metabolites in Healthy Control Subjects, Healthy Smokers, and COPD Patients Chest, February 1, 2009; 135(2): 353 - 367. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Barreto, M. P. Villa, C. Olita, S. Martella, G. Ciabattoni, and P. Montuschi 8-Isoprostane in Exhaled Breath Condensate and Exercise-Induced Bronchoconstriction in Asthmatic Children and Adolescents Chest, January 1, 2009; 135(1): 66 - 73. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Ckless, A. Lampert, J. Reiss, D. Kasahara, M. E. Poynter, C. G. Irvin, L. K. A. Lundblad, R. Norton, A. van der Vliet, and Y. M. W. Janssen-Heininger Inhibition of Arginase Activity Enhances Inflammation in Mice with Allergic Airway Disease, in Association with Increases in Protein S-Nitrosylation and Tyrosine Nitration J. Immunol., September 15, 2008; 181(6): 4255 - 4264. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. L. Borrill, K. Roy, and D. Singh Exhaled breath condensate biomarkers in COPD Eur. Respir. J., August 1, 2008; 32(2): 472 - 486. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Nadeem, A. Masood, and N. Siddiqui Review: Oxidant--antioxidant imbalance in asthma: scientific evidence, epidemiological data and possible therapeutic options Therapeutic Advances in Respiratory Disease, August 1, 2008; 2(4): 215 - 235. [Abstract] [PDF]
|
|
|
|
|
|
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]
|
|
|
|
 |
|
 S. Saito, A. Yamamoto-Katou, H. Yoshioka, N. Doke, and K. Kawakita Peroxynitrite Generation and Tyrosine Nitration in Defense Responses in Tobacco BY-2 Cells Plant Cell Physiol., June 1, 2006; 47(6): 689 - 697. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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]
|
|
|
|
|
|
A. Baydur Not All That Comes Out Is Hot Air Chest, May 1, 2005; 127(5): 1482 - 1485. [Full Text] [PDF]
|
|
|
|
|
|
K. Kostikas, M. Gaga, G. Papatheodorou, T. Karamanis, D. Orphanidou, and S. Loukides Leukotriene B4 in Exhaled Breath Condensate and Sputum Supernatant in Patients With COPD and Asthma Chest, May 1, 2005; 127(5): 1553 - 1559. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 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]
|
|
|
|
 |
|
 P Cap, J Chladek, F Pehal, M Maly, V Petru, P J Barnes, and P Montuschi Gas chromatography/mass spectrometry analysis of exhaled leukotrienes in asthmatic patients Thorax, June 1, 2004; 59(6): 465 - 470. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. D. Moloney, S. E. Mumby, R. Gajdocsi, J. H. Cranshaw, S. A. Kharitonov, G. J. Quinlan, and M. J. Griffiths Exhaled Breath Condensate Detects Markers of Pulmonary Inflammation after Cardiothoracic Surgery Am. J. Respir. Crit. Care Med., January 1, 2004; 169(1): 64 - 69. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. L. David, I. Romieu, J. J. Sienra-Monge, W. J. Collins, M. Ramirez-Aguilar, B. E. del Rio-Navarro, N. I. Reyes-Ruiz, R. W. Morris, J. M. Marzec, and S. J. London Nicotinamide Adenine Dinucleotide (Phosphate) Reduced:Quinone Oxidoreductase and Glutathione S-Transferase M1 Polymorphisms and Childhood Asthma Am. J. Respir. Crit. Care Med., November 15, 2003; 168(10): 1199 - 1204. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Sandrini, I. M. Ferreira, C. Gutierrez, J. R. Jardim, N. Zamel, and K. R. Chapman Effect of Montelukast on Exhaled Nitric Oxide and Nonvolatile Markers of Inflammation in Mild Asthma Chest, October 1, 2003; 124(4): 1334 - 1340. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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]
|
|
|
|
|
|
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]
|
|
|
|
|
|
E Baraldi, S Carraro, R Alinovi, A Pesci, L Ghiro, A Bodini, G Piacentini, F Zacchello, and S Zanconato Cysteinyl leukotrienes and 8-isoprostane in exhaled breath condensate of children with asthma exacerbations Thorax, June 1, 2003; 58(6): 505 - 509. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Nabeyrat, G. E. Jones, P. S. Fenwick, P. J. Barnes, and L. E. Donnelly Mitogen-activated protein kinases mediate peroxynitrite-induced cell death in human bronchial epithelial cells Am J Physiol Lung Cell Mol Physiol, June 1, 2003; 284(6): L1112 - L1120. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E Baraldi, L Ghiro, V Piovan, S Carraro, F Zacchello, and S Zanconato Safety and success of exhaled breath condensate collection in asthma Arch. Dis. Child., April 1, 2003; 88(4): 358 - 360. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Vass, E. Huszar, E. Barat, M. Valyon, D. Kiss, I. Penzes, M. Augusztinovicz, and I. Horvath Comparison of Nasal and Oral Inhalation during Exhaled Breath Condensate Collection Am. J. Respir. Crit. Care Med., March 15, 2003; 167(6): 850 - 855. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L.G. Wood, P.G. Gibson, and M.L. Garg Biomarkers of lipid peroxidation, airway inflammation and asthma Eur. Respir. J., January 1, 2003; 21(1): 177 - 186. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Paredi, S. A. Kharitonov, and P. J. Barnes Analysis of Expired Air for Oxidation Products Am. J. Respir. Crit. Care Med., December 15, 2002; 166(12): S31 - 37. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. P. Bowler and J. D. Crapo Oxidative Stress in Airways: Is There a Role for Extracellular Superoxide Dismutase? Am. J. Respir. Crit. Care Med., December 15, 2002; 166(12): S38 - 43. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Csoma, S. A. Kharitonov, B. Balint, A. Bush, N. M. Wilson, and P. J. Barnes Increased Leukotrienes in Exhaled Breath Condensate in Childhood Asthma Am. J. Respir. Crit. Care Med., November 15, 2002; 166(10): 1345 - 1349. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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]
|
|
|
|
|
|
L. Ghiro, S. Zanconato, O. Rampon, V. Piovan, M.F. Pasquale, and E. Baraldi Effect of montelukast added to inhaled corticosteroids on fractional exhaled nitric oxide in asthmatic children Eur. Respir. J., September 1, 2002; 20(3): 630 - 634. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Antczak, P. Montuschi, S. Kharitonov, P. Gorski, and P. J. Barnes Increased Exhaled Cysteinyl-Leukotrienes and 8-Isoprostane in Aspirin-induced Asthma Am. J. Respir. Crit. Care Med., August 1, 2002; 166(3): 301 - 306. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
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]
|
|
|
|
|
|
M. J. TOBIN Asthma, Airway Biology, and Allergic Rhinitis in AJRCCM 2000 Am. J. Respir. Crit. Care Med., November 1, 2001; 164(9): 1559 - 1580. [Full Text] [PDF]
|
|
|
|
|
|
C. E. Cross, A. van der Vliet, and J. P. Eiserich Peroxidases Wheezing Their Way into Asthma Am. J. Respir. Crit. Care Med., October 1, 2001; 164(7): 1102 - 1103. [Full Text] [PDF]
|
|
|
|
|
|
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]
|
|
|
|
|
|
S. A. KHARITONOV and P. J. BARNES Exhaled Markers of Pulmonary Disease Am. J. Respir. Crit. Care Med., June 1, 2001; 163(7): 1693 - 1722. [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| Proc. Am. Thorac. Soc. | Am. J. Respir. Cell Mol. Biol. |