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First published online on June 15, 2007
Chest, doi:10.1378/chest.07-0215
doi:10.1378/chest.07-0215
(Chest. 2007; 132:589-596)
© 2007 American College of Chest Physicians
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Eicosanoids in Exhaled Breath Condensate and BAL Fluid of Patients With Sarcoidosis*

Wojciech J. Piotrowski, MD, PhD; Adam Antczak, MD, PhD; Jerzy Marczak, MD, PhD; Agnieszka Nawrocka, MD, PhD; Zofia Kurmanowska, MSc and Pawel Górski, MD, PhD

* From the Division of Pneumonology and Allergy, (Drs. Piotrowski, Antczak, Marczak, and Górski, and Ms. Kurmanowska) and the Department of Pathology (Dr. Nawrocka), Medical University of Lodz, Lodz, Poland.

Correspondence to: Pawel Górski, MD, PhD, Division of Pneumonology and Allergy, Medical University of Lodz, 22 Kopciñskiego Str 90, 153 Lodz, Poland; e-mail: p_gorski{at}toya.net.pl

Abstract

Background: Measurement of inflammatory mediators in exhaled breath condensate (EBC) is an easy and noninvasive diagnostic method, which has gained popularity in the past few years. However, the source of these mediators is not precisely defined. It has been only presumed that inflammatory cells present in the airway lumen are the main source. Therefore, the aim of this study was to verify the relationship between EBC and BAL fluid (BALF) eicosanoids, and the percentage, number, and activity of cells in BALF.

Methods: In 28 sarcoidosis patients and 17 healthy subjects, 8-isoprostane, cysteinyl leukotrienes (CysLTs), and leukotriene B4 (LTB4) were measured in EBC by enzyme immunoassay. Eicosanoids were also examined in BALF in the study group. Cell count, percentage, and superoxide production by BALF cells were estimated.

Results: The mean (± SEM) CysLT and 8-isoprostane concentrations were higher in the sarcoidosis group (6.5 ± 0 vs 27.82 ± 6.65 pg/mL, respectively; and 2.67 ± 0.16 vs 13.95 ± 2.59 pg/mL, respectively). There were positive correlations between EBC and BALF 8-isoprostane concentration (r = 0.68, p < 0.0001) and LTB4 concentration (r = 0.43; p = 0.026). EBC LTB4 levels correlated with the number of lymphocytes per milliliter of BALF. The percentage and number of eosinophils in BALF correlated with EBC 8-isoprostane and BALF CysLT concentrations. No positive correlation was found between concentrations of EBC eicosanoids and percentages BALF lymphocytes, BALF macrophages, or superoxide production.

Conclusions: The levels of 8-isoprostane and CysLT are elevated in EBC in sarcoidosis patients; however, a lack of correlation with BALF lymphocyte percentage does not encourage us to recommend the measurement of eicosanoids as activity markers. The positive correlation of EBC 8-isoprostane and BALF CysLT concentrations with the percentage of eosinophils in BALF, and the higher percentage of eosinophils in BALF from patients with grade 3 sarcoidosis, may suggest the possible prognostic value.

Key Words: BAL fluid • eicosanoids • exhaled breath condensate • 8-isoprostane • leukotrienes • sarcoidosis

Exhaled breath condensate (EBC) may be useful in the monitoring of inflammation in the lung. The levels of thiobarbituric acid-reactive products, hydrogen peroxide (H2O2), nitrogen-derived oxidant species, and other oxidative stress metabolites are elevated in patients with various respiratory diseases.1 Other inflammatory mediators, including leukotriene B4 (LTB4), cysteinyl leukotrienes, and other eicosanoids have been used to assess inflammation.23 8-Isoprostane, a prostaglandin-F2-like compound that is produced in vivo by the free radical-catalyzed peroxidation of arachidonic acid, has been proposed as a reliable marker of oxidative stress.45678

BAL enables the assessment of inflammation directly from the airways. It is especially valuable in the differentiation of alveolitis.910 The presence of many inflammatory mediators can be measured in BAL fluid (BALF).1112 However, due to its invasiveness, bronchoscopy cannot be performed in all patients. Therefore, the measurement of EBC seems to be interesting as a noninvasive method for estimating inflammation in the lung.

The levels of H2O2 and thiobarbituric acid-reactive substances were only slightly elevated in the EBC of sarcoidosis patients.13 Elevated 8-isoprostane levels were found in the BALF6 and EBCs14 of these patients. Leukotrienes may play an important role in the regulation of chronic inflammation and fibrosis.151617 Increased production of LTB4 by macrophages isolated from the BALF of sarcoidosis patients was found,18 but have not been measured in EBC. Similarly, cysteinyl leukotriene (CysLT) levels in BALF and EBC have not been assessed in sarcoidosis patients.

It is generally assumed that oxidative stress metabolites are generated by activated inflammatory cells that are present in different compartments of the respiratory system, especially the airway lumen, as other inflammatory mediators. However, published data on the possible sources of mediators detected in EBC are minimal.

Therefore, we wanted to assess the following: (1) the differences in levels of eicosanoids (ie, CysLT, LTB4, and 8-isoprostane) in EBC of sarcoidosis patients and healthy subjects; (2) correlations between levels of eicosanoids in EBC, cells in BALF, superoxide production by BALF cells, and eicosanoid levels in BALF in patients with sarcoidosis.

Materials and Methods

Study Population
Bronchofiberoscopy with BAL was performed in 28 patients (10 women; mean [± SEM] age, 39.18 ± 2.00 years). Pulmonary sarcoidosis stage 1 was diagnosed in 8 patients, stage 2 was diagnosed in 10 patients, and stage 3 was diagnosed in 10 patients. The group was also divided according to the percentage of lymphocytes found in the BALF (ie, ≥ 18% and ≤ 17%).14 Sarcoidosis was confirmed by histopathologic examination in all patients (video-assisted thoracoscopy, five patients; transbronchial lung biopsy, nine patients; bronchial biopsy, 9 patients), except for five patients with stage 1 disease, in whom the diagnosis was based on clinical and radiologic criteria.19 The longest duration of clinical/radiologic symptoms was 3 months. Other respiratory disorders, especially bronchial tumor, asthma, COPD, bronchiectases, tuberculosis, acute infection (4 weeks), and atopy were excluded. None of the patients had ever smoked, and all were treatment-naive.

The control group consisted of 17 healthy, nonatopic never-smokers (8 women; mean [± SEM] age, 39.82 ± 2.62 years). Bronchoscopy and BAL were not performed in the control group. The protocol was approved by the local ethics committee at the Medical University of Lodz.

Lung Function Tests
Spirometry was performed according to American Thoracic Society (ATS)/European Respiratory Society (ERS) standards,20 on a computer-based spirometer (Jaeger; Hochberg, Germany). FVC, FEV1, and maximal midexpiratory flow were measured. The Tiffenau index (ie, FEV1/FVC ratio) was calculated. The results of the measurement of FVC, FEV1, and maximal midexpiratory flow were presented in liters and as the percentage of predicted values.21 The diffusing capacity of the lung for carbon monoxide was measured (Lungtest 1000 SB; MES; Krakow, Poland) with a single-breath method, according to ATS/ERS standards.22 The results of the measurement of diffusing capacity of the lung for carbon monoxide were corrected for hemoglobin concentration (DLCOc). Values were presented as the percentage of predicted values.

Bronchoscopy was performed with a flexible bronchoscope (Pentax; Tokyo, Japan) according to British Thoracic Society guidelines.23 BALF was collected from the medial lobe or lingula by the instillation and subsequent withdrawal of 4 x 50 mL portions of a 0.9% NaCl solution. The mean recovered portion of the instilled fluid was 52 ± 14%. The crude BALF was filtered through gauze and centrifuged, and the pellet was suspended in a phosphate buffer. The total cell count (TCC) was calculated (x106). Cytospin slides were stained with May-Grünwald-Giemsa stain. The numbers of macrophages, lymphocytes, neutrophils, and eosinophils were calculated under a light microscope, and were presented as a percentage of the TCC. In addition, the total number of cells and cell types were presented as the number of cells (x104) per milliliter of recovered fluid.

Superoxide anion (O2-.) production by BALF cells was measured colorimetrically, as previously described.24 The method is based on the reduction of cytochrome C by a O2-.. Superoxide dismutase was used to inhibit the reaction. Cells were stimulated with phorbol myristate acetate (1 ng/mL), and the difference between stimulated and spontaneous release of cells was taken for calculation of the concentration. Values were expressed in nanomoles of O2-. per 106 cells.

Collection of EBC
EBC was collected from patients and healthy control subjects using a commercial device (Ecoscreen; Jaeger). Patients were asked to breath out spontaneously for 10 min through a mouthpiece equipped with a saliva trap. The respiratory rate ranged from 15 to 20 breaths/min. All subjects wore a nose clip and rinsed their mouths with distilled water just before and in the seventh minute of collection in order to reduce nasal contamination. Samples were stored at –80°C. The collection of EBC was performed following available recommendations25 and always before bronchoscopy was performed.26

Measurement of Immunoreactive Eicosanoids
CysLT, LTB4, and 8-isoprostane concentrations in EBC and BALF were measured by a specific enzyme immunoassay kit (Cayman Chemical; Ann Arbor, MI), as previously described.2 The detection limits were 13 pg/mL for CysLT, 4.43 pg/mL for LTB4, and 5 pg/mL for 8-isoprostane. Levels of measured mediators below the detection limit were arbitrarily assumed to be half of the detection limit value.2 CysLT was measured in 25 EBC samples only, due to an insufficient volume of EBC (the measurement was not performed in two patients with stage 1 sarcoidosis and in one patient with stage 3 sarcoidosis).

Statistical Analysis
Data were expressed as the mean ± SEM. Median values with 25th and 75th percentiles were also provided when the data were not normally distributed. The Wilcoxon signed rank test with a theoretical median equal to the median of a control group was applied for the calculation of differences in EBC CysLT levels for the control and sarcoidosis groups (all data in the control group were equal). In all other instances, an unpaired t test (for normally distributed data) or Mann-Whitney U test (for data without normal distribution) was used. When more than two groups were compared (ie, patients in different stages of sarcoidosis), a one-way analysis of variance and posttest Bonferroni correction (for data with Gaussian distribution) or the Kruskal-Wallis test followed by Dunn multiple comparison test (for data without normal distribution) were used. The Spearman test was applied to assess correlations. A p value of ≤ 0.05 was assumed to be statistically significant.

Results

No statistical differences were found between the sarcoidosis and control groups in age and lung function test parameters, except for statistically lower, but not clinically relevant, mean percent predicted FEV1 values in the sarcoidosis group (104.2 ± 2.69% predicted vs 92.65 ± 3.52% predicted, respectively; p = 0.04). The composition of BALF is presented in Table 1 . In nine patients (32%), the percentage of lymphocytes in BALF was < 18%. Of those patients with a low percentage of lymphocytes in BALF, 78% had stage 2 and 3 sarcoidosis (22% and 56% of the subgroup, respectively). The number and percentage of eosinophils was higher in patients with stage 3 disease compared to those with stage 1 disease, but there were no differences in other cells found in BALF (Table 2 ). There were no differences in lung function test parameters between subgroups of patients with low and high percentages of lymphocytes in BALF, and between subgroups of patients in different stages of disease. The mean DLCOc was lower in stage 3 patients (73.99 ± 6.37% predicted) compared to stage 1 patients (98.28 ± 2.44% predicted; p < 0.05). DLCOc was not different between subgroups of patients with high and low percentages of lymphocytes in BALF.


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  		Table 1.. Composition of BALF in Patients With Sarcoidosis*

 

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  		Table 2.. Characteristics and Results of Measurements in Sarcoidosis Patients in Relation to Disease Stages*

 
The production of O2-. by cells in BALF was not different between patients in different disease stages (Table 2) or between patients with low and high percentages of lymphocytes in BALF (Table 3 ); the mean was 9.04 ± 2.3 nmol O2-. per 106 cells in the whole group (median, 0; 25th percentile, –0; 75th percentile, 19.32). In 54% of patients, BALF cells did not produce detectable levels of O2-. after stimulation with phorbol myristate acetate.


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  		Table 3.. Characteristics and Results of Measurements in Sarcoidosis Patients in Relation to Disease Stages*

 
The mean level of LTB4 in EBC was not different between groups (control subjects: mean, 20.09 ± 4.85 pg/mL; median, 15.44 pg/mL; 25th percentile, 3.61; 75th percentile, 30.78 pg/mL; sarcoidosis patients: mean, 23.14 ± 3.35 pg/mL; median, 21.05 pg/mL; 25th percentile, 6.73 pg/mL; 75th percentile, 36.4 pg/mL; p = 0.49). Four of 17 control subjects (24%) and 6 of 28 sarcoidosis patients (21%) had LTB4 concentrations below the detection limit. The mean CysLT level was higher in sarcoidosis patients (6.5 ± 0 vs 27.82 ± 6.65 pg/mL; median, 16.30 pg/mL; 25th percentile, 6.50 pg/mL; 75th percentile, 32.45 pg/mL; p < 0.0001) [Fig 1, top, A]. All subjects in a control group had CysLT levels below the detection limit, compared to 7 of 28 sarcoidosis patients (25%). The mean 8-isoprostane concentration EBC was also higher in the sarcoidosis group (13.95 ± 2.59 pg/mL; median, 8.35 pg/mL; 25 th percentile, 2.5 pg/mL; 75th percentile, 19.3 pg/mL) than in the control group (2.67 ± 0.16 pg/mL; median, 2.5 pg/mL; 25th percentile, 2.5 pg/mL; 75th percentile, 2.5 pg/mL; p = 0.0003) [Fig 1, bottom, B]. Sixteen of 17 control subjects (94%) had 8-isoprostane concentrations below the detection limit, compared to 9 of 28 sarcoidosis patients (32%).


Figure 1
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  		Figure 1.. Top, A: comparison of CysLT concentrations in the EBC of patients with sarcoidosis (n = 25) and a control group (n = 17). The thin horizontal line describes the detection limit (13 pg/mL). Bottom, B: comparison of 8-isoprostane levels in the EBC of patients with sarcoidosis (n = 28) and a control group (n = 17). The thin horizontal line describes the detection limit (5 pg/mL).

 
The concentrations of eicosanoids in BALF were not different between patients in different sarcoidosis stages (Table 2) and between subgroups of patients with low and high percentages of lymphocytes (Table 3). The concentrations of eicosanoids in BALF in the whole group are presented in Table 1.

Correlations were found between eicosanoid concentrations in EBC and BALF (8-isoprostane, Fig 2 top, A; LTB4, Fig 2, bottom, B). All significant correlations between eicosanoids in EBC and BALF and cellular components are presented in Table 4 .


Figure 2
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  		Figure 2.. Top, A: correlation between LTB4 concentrations in EBC and BALF in patients with sarcoidosis (n = 28). Bottom, B: correlation between 8-isoprostane levels in EBC and BALF in patients with sarcoidosis (n = 28).

 

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  		Table 4.. Statistically Significant Correlations Between Eicosanoids in BALF and EBC and Cells in BALF

 
There were no correlations between DLCOc and eicosanoid concentrations in EBC. The negative correlation was found among DLCOc percent predicted, and eosinophil and CysLT levels in BALF (Table 4).

Discussion

Our study confirms the presence of elevated levels of 8-isoprostane in the EBC of patients with sarcoidosis. To our knowledge, this is the first study to show increased concentrations of CysLT in the EBC of these patients.

The increased production of hydrogen peroxide by BALF macrophages and the presence of oxidized BALF proteins were found previously in sarcoidosis patients.2728 Elevated concentrations of 8-isoprostane in EBC14 and BALF6 have been reported by other authors. In the cited study,6 there were no correlations found between 8-isoprostane levels and BALF cell types. Psathakis et al14 found the positive correlation with serum concentrations of angiotensin-converting enzymes, but not with serum calcium levels and pulmonary function test parameters.

Increased concentrations of LTB4 in EBC have been found in patients in different disease states with significant neutrophil involvement.2930 LTB4 can also be released by activated alveolar macrophages in sarcoidosis patients.18 Gaber et al31 reported that the main source of LTB4 detected in EBC was saliva. We have found a positive correlation between EBC and BALF LTB4 concentrations, which stands in opposition to the cited authors’ conclusions.31

So far, elevated CysLT levels have been reported in patients with asthma,32 where mastocytes and eosinophils are its main sources. In our study, we found elevated concentrations of CysLT in EBC. Levels of CysLT in BALF also correlated with the percentage and number of eosinophils per milliliter of BALF. We have also found a negative correlation between DLCOc and the percentage and number of eosinophils in BALF and the level of CysLT in BALF. However, we have not found a correlation between CysLT levels in EBC and BALF.

There is common agreement that the most probable source of these mediators is BALF cells. Neutrophils and eosinophils are the most potent cellular sources of reactive oxygen species on a per-cell basis. The positive correlation between 8-isoprostane levels in EBC and the numbers and percentages of eosinophils in BALF may confirm this observation. Taking into account the higher eosinophil percentage in patients with stage 3 sarcoidosis compared to those with stage 1 sarcoidosis, and the negative correlation between eosinophils and DLCOc, this observation is consistent with the report of Ziegenhagen et al,33 who found that a higher percentage of eosinophils (and neutrophils) in BALF in newly diagnosed sarcoidosis patients was associated with a significantly higher risk of necessity for steroid therapy, and may be a good marker of progressive disease. Macrophages are thought to be the most probable cellular source of oxidative stress markers and other inflammatory mediators, due both to the per-cell potential and the prevalence in BALF. But, an increased percentage of lymphocytes in BALF is one of best predictors of disease activity19; an increased percentage of lymphocytes always results in a decreased percentage of macrophages. Therefore, the activity of macrophages should increase several-fold in order to cause elevated concentrations of these mediators in EBC. In our study, we not only found a lack of positive correlation with lymphocytes, but also no correlation with both the number of cells per milliliter and the percentage of macrophages in BALF, and, most importantly, with superoxide production by BALF cells.

Besides the above-mentioned correlations, we have found positive correlations between BALF and EBC eicosanoids (ie, 8-isoprostanes and LTB4). Jackson et al34 measured the levels of 8-isoprostane (and other mediators) in a heterogeneous group of patients, and they found no correlation between biomarkers in EBC and those in BALF. The authors compared 8-isoprostane levels in EBC and BALF, both of which were standardized for protein concentration. In our study, we have used data that were standardized for the volume of recovered fluid for all calculations. This way of presenting data referred to all of the correlated values (ie, EBC/BALF eicosanoids and cellular components in BALF). Protein, albumin, and urea concentrations in BALF may be increased when cell permeability is increased, for instance, in patients with chronic inflammation and sarcoidosis.35 The use of correction markers is even more doubtful in EBC, as the source of mediators is not only alveolar space, and eicosanoids and dilution markers have completely different physical properties. Correction for dilution is also not recommended by the ERS/ATS task force on EBC.25 Also, ERS guidelines36 for the measurement and standardization of acellular BALF components state that the empirical expression of BALF results as "per milliliter of aspirate" remains the most popular way of expressing data since it avoids potentially confounding assumptions. However, it is worth adding that when we standardized our concentrations of eicosanoids in BALF, either for albumin or urea, the correlation between 8-isoprostane levels in BALF and EBC, and the majority of other correlations were preserved (data not shown). As a result, the following two questions arise: (1) what is the real cellular source of mediators detected both in EBC and BALF?; and (2) can these EBC mediators be used as predictors of disease activity?

These inconsistent data should stimulate the search for other potential sources of EBC mediators. Oxidative stress metabolites and other inflammatory mediators may be released by the alveolar epithelium. Type II pneumocytes produce nitric oxide, eicosanoids, and other mediators.373839 They may be stimulated to produce oxidants.40 Their membrane lipids may undergo peroxidation when exposed to oxidative stress.41 They are progenitors of type I pneumocytes.42 These data gain special power when we consider the huge area of alveoli covered by alveolar cells.

In conclusion, although we found elevated concentrations of 8-isoprostane and CysLT in the EBCs of sarcoidosis patients and some correlations of 8-isoprostane levels in EBC with the number and percentage of eosinophils in BALF, due to the lack of correlations with the percentage of lymphocytes in BALF these biomarkers should not be recommended as predictors of disease activity. Further studies on the source and reliability of these EBC mediators for the estimation of inflammation and oxidative stress in the lung are needed. In view of our results, it will be especially interesting to verify the usefulness of eicosanoids in EBC as prognostic markers.

Footnotes

Abbreviations: ATS = American Thoracic Society; BALF = BAL fluid; CysLT = cysteinyl leukotriene; DLCOc = diffusing capacity of the lung for carbon monoxide corrected for hemoglobin concentration; EBC = exhaled breath condensate; ERS = European Respiratory Society; LTB4 = leukotriene B4; TCC = total cell count

This study was supported by Medical University of Lódz individual grants No. 502-11-048 and 502-11-451 (to Dr. Piotrowski).

The authors have reported to the ACCP that no significant conflicts of interest exist with any companies/organizations whose products or services may be discussed in this article.

Received for publication January 24, 2007. Accepted for publication April 16, 2007.

References

  1. Kharitonov, SA, Barnes, PJ (2006) Exhaled biomarkers. Chest 130,1541-1546[Abstract/Free Full Text]
  2. Antczak, A, Montuschi, P, Kharitonov, S, et al Increased exhaled cysteinyl-leukotrienes and 8-isoprostane in aspirin-induced asthma. Am J Respir Crit Care Med 2002;165,1-6[Free Full Text]
  3. Mondino, C, Ciabattoni, G, Koch, P, et al Effects of inhaled corticosteroids on exhaled leukotrienes and prostanoids in asthmatic children. J Allergy Clin Immunol 2004;114,761-767[CrossRef][ISI][Medline]
  4. Montuschi, P, Corradi, M, Ciabattoni, G, et al Increased 8-isoprostane, a marker of oxidative stress, in exhaled condensate of asthma patients. Am J Respir Crit Care Med 1999;16,216-220
  5. Antczak, A, Kharitonov, SA, Montuschi, P, et al Inflammatory response to sputum induction measured by exhaled markers. Respiration 2005;26,523-548
  6. Montuschi, P, Ciabattoni, G, Paredi, P, et al 8-Isoprostane as a biomarker of oxidative stress in interstitial lung diseases. Am J Respir Crit Care Med 1998;158,1524-1527[Abstract/Free Full Text]
  7. Montuschi, P, Kharitonov, SA, Ciabattoni, G, et al Exhaled 8-isoprostane as a new non-invasive biomarker of oxidative stress in cystic fibrosis. Thorax 2000;55,205-209[Abstract/Free Full Text]
  8. Montuschi, P, Collins, JV, Ciabattoni, G, et al Exhaled 8-isoprostane as an in vivo biomarker of lung oxidative stress in patients with COPD and healthy smokers. Am J Respir Crit Care Med 2000;162,1175-1177[Abstract/Free Full Text]
  9. Hunninghake, GW, Crystal, RG Pulmonary sarcoidosis: a disorder mediated by excess helper T lymphocyte activity at sites of disease activity. N Engl J Med 1981;305,429-434[Abstract]
  10. Costabel, U, Bross, JK, Marxen, J, et al T lymphocytes in broncholaveolar lavage fluid of hypersensitivity pneumonitis: changes in profile of T cell subsets during the course of the disease. Chest 1984;85,514-518[Abstract/Free Full Text]
  11. Suga, M, Iyonaga, K, Ichiyasu, N, et al Clinical significance of MCP-1 levels in BALF and serum in patients with interstitial lung disease. Eur Respir J 1999;14,376-382[Abstract]
  12. Moodley, YP, Dorasamy, T, Venketasamy, S, et al Correlation of CD4: CD8 ratio and tumour necrosis factor (TNF) {alpha} levels in induced sputum with bronchoalveolar lavage fluid in pulmonary sarcoidosis. Thorax 2000;55,696-699[Abstract/Free Full Text]
  13. Kwiatkowska, S, Luczyñska, M, Grzelewska-Rzymowska, I, et al Comparison of oxidative stress markers in exhaled breath condensate and in serum of patients with tuberculosis and sarcoidosis. Pol Merkur Lekarski 2005;19,37-40[Medline]
  14. Psathakis, K, Papatheodorou, G, Plataki, M, et al 8-isoprostane, a marker of oxidative stress, is increased in the expired breath condensate of patients with pulmonary sarcoidosis. Chest 2004;125,1005-1011[Abstract/Free Full Text]
  15. Nieves, D, Moreno, JJ Role of 5-lipoxygenase pathway in the regulation of RAW 264.7 macrophage proliferation. Biochem Pharmacol 2006;72,1022-1030[CrossRef][ISI][Medline]
  16. Serezani, CH, Aranoff, DM, Jancar, S, et al Leukotrienes enhance the bactericidal activity of alveolar macrophages against Klebsiella pneumoniae through the activation of NADPH oxidase. Blood 2005;106,1067-1075[Abstract/Free Full Text]
  17. Beller, TC, Friend, DC, Maekawa, A, et al Cysteinyl leukotriene 1 receptor controls the severity of chronic pulmonary inflammation and fibrosis. Proc Natl Acad U S A 2004;101,3047-3052[Abstract/Free Full Text]
  18. De Rose, V, Trentin, L, Crivellari, MT, et al Release of prostaglandin E2 and leukotriene B4 by alveolar macrophages from patients with sarcoidosis. Thorax 1997;52,76-83[Abstract/Free Full Text]
  19. Statement on sarcoidosis.. The joint statement of ATS, ERS and WASOG. Am J Respir Crit Care Med 1999;160,736-755[Free Full Text]
  20. American Thoracic Society.. Standardization of spirometry, 1994 update. Am J Respir Crit Care Med 1995;152,1107-1136[ISI][Medline]
  21. Miller, MR, Hankinson, J, Brusasco, V, et al ATS/ERS Task Force: standardisation of lung function testing; standardisation of spirometry. Eur Respir J 2005;26,319-338[Abstract/Free Full Text]
  22. Pellegrino, R, Viegi, G, Brusasco, V, et al ATS/ERS Task Force: standardisation of lung function testing; interpretative strategies for lung function tests. Eur Respir J 2005;26,948-968[Free Full Text]
  23. British Thoracic Society guidelines on diagnostic flexible bronchoscopy.. British Thoracic Society Bronchoscopy Guidlines Committee, a Subcommittee of the Standards of Care Committee of the British Thoracic Society. Thorax 2001;56(suppl),i1-i21[Free Full Text]
  24. Suzuki, Y, Lehrer, RI NAD(P)H oxidase activity in human neutrophils stimulated by phorbol myristate acetate. J Clin Invest 1980;66,1409-1418[ISI][Medline]
  25. Horvath, I, Hunt, J, Barnes, PJ Exhaled breath condensate: methodological recommendations and unresolved questions. Eur Respir J 2005;26,523-548[Abstract/Free Full Text]
  26. Piotrowski, WJ, Antczak, A, Marczak, J, et al The influence of bronchoscopy and bronchoalveolar lavage (BAL) on cysteinyl leukotrienes and 8-isoprostanes in exhaled breath condensate [abstract].Am J Respir Crit Care Med 2002;165(suppl),A805
  27. Lenz, AG, Costabel, U, Maier, KL Oxidized BAL fluid proteins in patients with interstitial lung diseases. Eur Respir J 1996;9,307-312[Abstract]
  28. Fels, AOS, Nathan, CF, Cohn, ZA Hydrogen peroxide release by alveolar macrophages from sarcoid patients and by alveolar macrophages from normals after exposure to recombinant interferons {alpha}A, B, and {gamma} and 1,25-dihydroxyvitamin D3. J Clin Invest 1987;80,381-386[ISI][Medline]
  29. Ham, EA, Soderman, DD, Zanetti, ME, et al Inhibition by prostaglandins of leukotriene B4 release from activated neutrophils. Proc Natl Acad Sci U S A 1983;80,4349-4353[Abstract/Free Full Text]
  30. Montuschi, P, Kharitonov, SA, Ciabattoni, G, et al Exhaled leukotrienes and prostaglandins in COPD. Thorax 2003;58,585-588[Abstract/Free Full Text]
  31. Gaber, F, Acevedo, F, Delin, I, et al Saliva is one likely source of leukotriene B4 in exhaled breath condensate. Eur Respir J 2006;28,1229-1235[Abstract/Free Full Text]
  32. Cap, P, Chladek, J, Pehal, F, et al Gas chromatography/mass spectrometry analysis of exhaled leukotrienes in asthmatic patients. Thorax 2004;59,465-470[Abstract/Free Full Text]
  33. Ziegenhagen, MW, Rothe, ME, Schlaak, M, et al Bronchoalveolar and serological parameters reflecting the severity of sarcoidosis. Eur Respir J 2003;21,407-413[Abstract/Free Full Text]
  34. Jackson, AS, Sandrini, A, Campbell, C, et al Comparison of biomarkers in exhaled breath condensate and bronchoalveolar lavage. Am J Respir Crit Care Med 2007;175,222-227[Abstract/Free Full Text]
  35. Roberts, CM, Caims, D, Bryant, DH, et al Changes in epithelial lining fluid albumin associated with smoking and interstitial lung disease. Eur Respir J 1993;6,110-115[Abstract]
  36. Ward, C, Effros, RM, Walters, EH Assessment of epithelial lining fluid dilution during bronchoalveolar lavage. Eur Respir Rev 1999;9,32-37
  37. Pawliczak, R, Huang, XL, Nanvaty, UB, et al Oxidative stress induces arachidonate release from human lung cells through the epithelial growth factor receptor pathway. Am J Respir Cell Mol Biol 2002;27,722-731[Abstract/Free Full Text]
  38. Punjabi, CJ, Laskin, JD, Pendino, KJ, et al Production of nitric oxide by type II pneumocytes: increased expression of inducible nitric oxide synthase following inhalation of a pulmonary irritant. Am J Respir Cell Mol Biol 1994;11,165-172[Abstract]
  39. Grimminger, F, von Kurten, I, Walmrath, D, et al Type II alveolar epithelial eicosanoid metabolism: predominance of cyclooxygenase pathways and transcellular lipooxygenase metabolism in co-culture with neutrophils. Am J Respir Cell Mol Biol 1992;6,9-16[ISI][Medline]
  40. Piotrowski, WJ, Marczak, J, Dinsdale, D, et al Release of hydrogen peroxide by rat type II pneumocytes in the prolonged culture. Toxicol In Vitro 2000;14,85-93[CrossRef][ISI][Medline]
  41. Piotrowski, WJ, Marczak, J, Kurmanowska, Z, et al Concentration of TBA-reactive substances in type II pneumocytes exposed to oxidative stress. Arch Immunol Ther Exp 2004;52,435-440
  42. Paine, R, Gaposchkin, D, Kelly, C, et al Regulation of cytokeratin expression in rat lung alveolar epithelial cells in vitro. Am J Physiol 1995;269,L536-L544[ISI][Medline]




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