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Exhaled nitric oxide before and after montelukast sodium therapy in school-age children with chronic asthma: a preliminary study.

Exhaled nitric oxide (ENO) is a surrogate marker of airway inflammation in asthma. In 12 children aged 6-11 years with mild to moderate persistent asthma, ENO concentrations were measured before and after 4 weeks of treatment with montelukast sodium, a leukotriene receptor antagonist, and 2 weeks after withdrawal of therapy. Baseline ENO levels (mean and 95% confidence interval) were significantly elevated in patients with asthma compared to age-matched nonasthmatic control subjects, with levels of 83 (42-123) vs. 13 (11-15) ppb (P < 0.001). After treatment with montelukast sodium, there was a significant (P < 0.01) reduction in ENO to 58 (27-89) ppb which again rose to 69 (38-99) ppb 2 weeks after treatment was withdrawn. During treatment, the fall in ENO was accompanied by nonsignificant improvements in prebronchodilator forced expiratory volume in 1 s (FEV(1)) from 81-85% predicted or reductions in use of albuterol from a mean of 2.5 to 1.6 puffs/day. Individual ENO measurements and change in ENO concentrations with treatment did not correlate with either pulmonary function changes or use of bronchodilator. These data show that ENO is elevated in children with relatively mild asthma treated with bronchodilator alone, and that treatment with montelukast sodium for 4 weeks results in a significant reduction in ENO concentrations, even in the absence of significant changes in pulmonary function. These findings suggest an anti-inflammatory role for leukotriene D(4) receptor antagonism in the treatment of children with mild to moderate asthma.

Acetates↗

Exhaled nitric oxide is reduced after sputum induction in asthmatic children.

Exhaled nitric oxide (ENO) and eosinophil sputum markers are considered noninvasive markers of airway inflammation in asthma. The aim of this study was to evaluate whether the procedure of sputum induction can affect the level of ENO. We measured ENO before and after sputum induction by inhalation of hypertonic saline solution in 22 asthmatic children and 9 healthy controls. The ENO mean (+/- S. E.M.) value in the group of asthmatic children was reduced from a baseline value of 20.8 (+/- 3.0) ppb to 17.4 (+/- 2.4) ppb after sputum induction (P = 0.0012). In the healthy controls, the mean baseline value of ENO was 9.1 (+/- 2.1) ppb and it was reduced to 4. 8 (+/- 1.1) ppb after induction of sputum (P < 0.01). We suggest that measurements of ENO should be performed after the induction of sputum in asthmatic patients whenever both tests are done in sequence.

Adolescent↗

Effect of 50 Hz sinusoidal electromagnetic field on the kinetics of 14CO2 exhalation after [14C]-N-nitrosodiethylamine administration in mice.

N-Nitrosodiethylamine (NDEA) has been identified as a typical environmental carcinogen. Its metabolism was studied in mice under the influence of an electromagnetic field (EMF). After intraperitoneal administration of [14C]-NDEA, 0.2 microCi/100 g body weight resulted in 22.8% of the total radioactivity exhaled as 14CO2 within 1 h. Mice were exposed to a 50 Hz, 2 mT (rms) electromagnetic field, 8 h/day for 8 weeks. There was a significant increase in the metabolic turnover of [14C]-NDEA into 14CO2 at the end of both 6 and 8 weeks of field exposure, i.e., 26.9% and 37.4% respectively. The enhanced capacity of mice to metabolize NDEA after the exposure to EMF may result in animals with a smaller amount of the bioactive carcinogen burden, thereby indicating a protective role of 2 mT EMF in a whole animal study.

Animals↗

Correlation between reversibility of airway obstruction and exhaled nitric oxide levels in children with stable bronchial asthma.

Recent trials measuring exhaled nitric oxide (eNO) concentrations have suggested that it may be a useful measure of ongoing airway inflammation in patients with asthma. The purpose of this study was to examine the relationship between eNO levels and baseline as well as postbronchodilator spirometry, a measurement commonly used in the clinical setting to determine the severity of asthma and as a guide to therapeutic decisions. Forty-nine patients between the ages of 5-16 years with physician-diagnosed asthma who attended the pediatric pulmonary clinic for a routine asthma visit with spirometric evaluation were recruited for the study. eNO levels prior to spirometry were obtained before and after receiving inhaled beta(2) agonist. eNO samples were collected in impermeable bags (Tedlar) and assayed within 24 hr by chemiluminescence. Regression analysis was used to assess the relationships between pre- and postbronchodilator eNO and spirometric variables. eNO was also compared in patients receiving and not receiving inhaled corticosteroids (ICS), as well as those whose therapy had been increased after evaluation by a pediatric pulmonologist or allergist. We found no significant difference between the levels of eNO before and after inhalation of beta(2) agonist (P = 0.60 paired t-test). Positive correlation was found between eNO vs. percentage change in FEV(1) (r = 0.35, P = 0.01) and percentage change in FEF(25-75% )(r = 0.29, P = 0.04). A negative correlation was found between prebronchodilator FEV(1) and eNO (r = -0.29, P = 0.03). Patients on ICS had lower mean eNO levels (29.9 vs. 47.6 parts per billion (ppb), P = 0.053) than those not receiving ICS. Patients whose ICS therapy was increased had higher mean eNO levels (47.2 vs. 26.9 ppb, P = 0.018) than those not having ICS therapy increased. We suggest that eNO levels could be a clinically useful measurement of asthma severity and could be used as an adjunct to spirometry to determine appropriate treatment plans. Longitudinal clinical trials are needed to determine if eNO can enhance therapeutic decisions for asthmatic children.

Adolescent↗

Protein pattern of exhaled breath condensate and saliva.

The proteins recovered in exhaled breath condensate (EBC) might be used to non-invasively monitor respiratory diseases. However, the range of proteins and their source are still unresolved and contamination by saliva or a similar protein pattern in the nasal and bronchial compartments may make interpretation of the data difficult. We studied nasal EBC (collected through a "free of touch" technique by negative pressure), oral tidal, and oral forced EBC (collected through a rebreathing valve as a saliva trap connected to tubing submerged into ice) and matched saliva samples from five healthy adult subjects. The protein samples were separated by two-dimensional electrophoresis and the silver stained gels were analyzed by Melanie 2 software. In both nasal and oral EBC, three spots (72 kDa/isoelectric point (pI) 6.6-7.0, 66 kDa/pI 5.9-6.7 and 45-48 kDa/pI 8.0-8.6) were consistently present in all subjects. Several other proteins were only sporadically detected. Despite improbable saliva contamination (no phosphorus contamination in the same oral and nasal EBC, no amylase activity in 10 pairs of nasal and oral EBC collected by the same technique), on average 63% and 71% of the spots identified in oral and nasal EBC were also found in the matched individual saliva samples. Compared to saliva, the range and amount of protein in all types of EBC was very small. Even when collected free of saliva contamination the majority of proteins present in EBC was also found in saliva, suggesting that these proteins are present in both compartments, e.g. saliva and secretions of the lower airspaces. The quantification and identification of specific proteins in the various compartments is warranted in future studies to determine the practical value of EBC.

Adult↗

Decreased exhaled nitric oxide in sickle cell disease: relationship with chronic lung involvement.

A deficiency in airway nitric oxide (NO) could contribute to pulmonary vaso-occlusion in sickle cell disease (SCD). We measured the fractional expired concentration of NO (FE(NO)) by chemiluminescence during a slow vital capacity maneuver against a positive pressure of 16 cm H(2)O at an expiratory flow rate of 50 mL/sec in 44 stable ambulatory adults with SCD and 30 healthy controls. A history of acute chest syndrome was present in 29 patients, and 22 complained of dyspnea. Mean +/- SD FE(NO) was significantly reduced in the SCD group compared with controls (14.8 +/- 8.4 vs. 24.9 +/- 13.5 ppb, P < 0.001). SCD patients with dyspnea had lower FE(NO) than those without dyspnea (10.1 +/- 5.7 vs. 19.6 +/- 8 ppb, P < 0.001) and those with a history of ACS had lower values than those no episodes of ACS (13.0 +/- 8.3 vs. 18.4 +/- 7.6 ppb, P < 0.05). There was a weak correlation between FE(NO) and percent-predicted DLCO (r = 0.4, P = 0.02) among the SCD patients. We conclude that exhaled NO is reduced in adults with SCD, and this may play a role in the pathogenesis of acute chest syndrome and chronic sickle cell lung disease.

Adult↗

Multiplex analysis of cytokines in exhaled breath condensate.

BACKGROUND: To improve monitoring of lung diseases, we analyzed cytokines in exhaled breath condensate (EBC). The main challenge in measurement of cytokines in EBC is the low protein content, which requires concentration steps that conflict with the need for excessive fluid required by most commonly used kits. METHODS: Here, a multiplex bead array for the detection of interleukins (IL) -1beta, -6, -8, -10, TNF-alpha, and IL-12p70 was modified and validated for analysis in EBC samples. Furthermore, 33 healthy volunteers and 11 patients with acute lung injury were investigated. RESULTS: In patients with inflammatory lung diseases, cytokine levels for all investigated cytokines were higher in comparison to healthy smokers or healthy volunteers. DISCUSSION: Multiplexed immunoassays in highly sensitive approaches allow for cytokine detection in EBC. We found significant differences between patients and controls for all investigated cytokines.

Adult↗

Correlation between concentrations of n-hexane and toluene in exhaled and environmental air in an occupationally exposed population.

We determined the correlations between the concentrations of n-hexane and toluene in exhaled and environmental air in the shoe manufacturing industry. Data were collected in 1988 and in 1992 from a total of 265 subjects. Environmental air samples were collected with personal diffusive samplers by adsorption on activated charcoal during exposure and from end-expired air (alveolar air) on cartridges of activated charcoal after exposure. Both compounds were desorbed with carbon disulphide and analysed by gas chromatography. Linear regression analyses showed a good correlation between environmental and end-expired air concentrations (r = 0.82 for n-hexane and r = 0.81 for toluene). These correlations allowed us to calculate the concentrations in expired air corresponding to current environmental limit values. The calculated concentrations in end-expired air that correspond to current environmental threshold limit values of 176 mg m-3 for n-hexane and 377 mg m-3 for toluene are 28 mg m-3 (95% confidence limit, 27-29 mg m-3) and 40 mg m-3 (95% confidence limit, 39-41 mg m-3), respectively. Similar correlations were found when the data from the two study periods were analysed separately.

Adolescent↗

The effect of acute pain crisis on exhaled nitric oxide levels in children with sickle cell disease.

Exhaled nitric oxide (FE(NO)) has been shown to be decreased in children with sickle cell disease. We sought to evaluate the effect of sickle cell vaso-occlusive crisis (VOC) on FE(NO) levels. We measured FE(NO) levels in 42 children with sickle cell disease, 29 in their baseline health and 13 during an acute VOC. There was no difference in FE(NO) levels between children at baseline (15.12 +/- 9.32 ppb) and those during an acute VOC (15.68 +/- 7.26 ppb; P = 0.794). FE(NO) is not a useful marker of acute VOC in children with sickle cell disease.

Acute Disease↗

Exhaled nitric oxide concentrations: online versus offline values in healthy children.

Exhaled nitric oxide (FE(NO)) is a noninvasive and practical method to assess airway inflammation. We conducted this investigation to determine the most appropriate flow rate to measure FE(NO) and to obtain reference values for FE(NO) in children. FE(NO) was measured in 112 healthy 6-18 year olds (60 males) at 4 expiratory flow rates (46, 31, 23, and 15 mL/sec) using a chemiluminescent nitric oxide analyzer. Offline and online analyses were done to determine FE(NO) intraclass correlation coefficients, the relationship between FE(NO) and expiratory flow rates, and the effects of age and gender on these measurements. The major findings were: 1) intraclass correlation coefficients for FE(NO) and flow rates ranged from 0.92-0.99 for offline values, and 0.99 for all online values; 2) variation at an expiratory flow rate of 46 mL/sec (SD, 9.39) was considerably less than at other flows, especially at 15 mL/sec (SD, 26.55); 3) FE(NO) increased as flow rates decreased for both offline and online values; 4) there were no significant differences and good agreement between offline bag and online FE(NO) values at 31 and 46 mL/sec expiratory flows; and 5) using multiple regression, significant predictors of FE(NO) were flow, body surface area, age, and FEF(25-75). We have provided FE(NO) values in healthy children and propose that the ideal expiratory flow rate for FE(NO) measurements in children using the single breath technique is between 30-50 mL/sec.

Adolescent↗

Exhaled nitric oxide at school age in prematurely born infants with neonatal chronic lung disease.

Prematurely born infants with neonatal chronic lung disease (CLD) have increased respiratory morbidity and bronchial obstruction at school age. To evaluate the possible inflammatory basis of lung function abnormalities, we studied 40 children, 7.5-9.6 years of age, born very prematurely (birth weights, 600-1,575 g) and 14 nonatopic term-born controls, using flow-volume spirometry and exhaled nitric oxide (eNO) measurements. In children born prematurely, eNO was significantly higher in atopics than in nonatopics (respective means, 14.8 vs. 6.3 ppb, P = 0.02). Nonatopic prematurely born infants did not differ significantly from controls (means, 6.3 vs. 6.4 ppb, P = ns). Of the 27 nonatopic children not on regular glucocorticoid inhalations, 9 had a history of CLD. Spirometry indicated bronchial obstruction and values that were significantly lower in prematurely born infants with or without CLD than in controls, and they were lower in the CLD than the non-CLD group. However, no significant differences were observed in eNO levels between CLD, non-CLD, and control groups (means, 6.8, 5.9, and 6.4 ppb, P = ns). In nonatopic schoolchildren born very prematurely and with a history of CLD, we found no evidence of airway inflammation associated with increased eNO concentrations. Neither were eNO levels associated with severity of chronic lung disease, as determined by conventional lung function tests. eNO levels were higher in atopic children born prematurely than in controls.

Child↗

Correlations between exhaled nitric oxide levels, blood eosinophilia, and airway obstruction reversibility in childhood asthma are detectable only in atopic individuals.

The aim of this study was to compare in atopic and nonatopic asthmatic children correlations between two inflammation parameters, i.e., blood eosinophilia and exhaled nitric oxide (FE(NO)), and pulmonary function values, at baseline and after beta(2)-adrenergic bronchodilators. Ninety-two steroid-naive asthmatic children were evaluated: 26 were skin prick test- and RAST-negative (nonatopic subjects), whereas 66 were atopic, 15 being sensitized only to house dust mites (monosensitized) and 51 to mites and to at least one other class of allergens (polysensitized). Baseline spirometric values (FEV(1) and FEF(25-75%)) were similar in atopic and nonatopic groups (P > 0.1, each comparison). However, when compared to nonatopic subjects, atopic children showed a significantly higher degree of blood eosinophilia (3.0% and 6.7% white blood cell count, respectively; P = 0.0001) and higher FE(NO) levels (6.8 ppb and 16.0 ppb, respectively; P = 0.0001). While a positive correlation between FE(NO) levels and blood eosinophilia was observed in atopic children (r = 0.25, P = 0.041), no correlations between these two inflammation parameters and baseline pulmonary function values were demonstrated in any of the asthmatic groups. Inhalation of a beta(2)-agonist drug induced in the two asthmatic populations similar improvements in FEV(1) and FEF(25-75%) and no changes in FE(NO) levels or blood eosinophilia. However, only in atopic children positive correlations were found between percent variation in FEV(1) (delta%FEV(1)) and FE(NO) levels (r = 0.35, P = 0.006) or blood eosinophilia (r = 0.26, P = 0.04). Within the atopic group, no differences were found between mono- and polysensitized individuals in all parameters evaluated. Thus only in atopic children did parameters of inflammation correlate with airway obstruction reversibility.

Adrenergic beta-Agonists↗

Exhaled nitric oxide corresponds with office evaluation of asthma control.

Exhaled NO (ENO) has been studied as a noninvasive marker of airway inflammation, and has been shown to be elevated in asthma patients. The aim of this study was to investigate whether ENO measurements differ significantly between groups of asthmatic children with different disease control and to compare ENO measurements with the clinical assessment of asthma control. Seventy-three children between 5-18 years old with a diagnosis of asthma were recruited. ENO was measured online during a slow vital capacity maneuver. The mean of three plateau NO levels was used for analysis. Baseline and postbronchodilator spirometry were performed. The assessment of disease control was based on the frequency of use of beta2-agonists, occurrence of day- and nighttime asthma symptoms, and spirometry results. Twenty-one children (group 1) had good asthma control. In 31 patients (group 2), asthma control was acceptable. In 21 patients (group 3), asthma was insufficiently controlled. ENO levels were (median (quartiles)): group 1, 11 ppb (9-21); group 2, 15 ppb (11-26); and group 3, 28 ppb (19-33). Measurements were significantly different between all three groups (P = 0.009, Kruskal-Wallis), between groups 1 and 3 (P = 0.01, Mann-Whitney U test), and between groups 2 and 3 (P = 0.01, Mann-Whitney-U test). The same was true for reversibility testing. We found significantly different ENO levels between a group of pediatric asthma patients with insufficient and good/sufficient control, as defined by clinical assessment. These results suggest that ENO measurements may be useful for monitoring asthma patients.

Albuterol↗

Effect of montelukast on time-course of exhaled nitric oxide in asthma: influence of LTC4 synthase A(-444)C polymorphism.

Leukotrienes (LT) mediate inflammation in asthma. The fraction of exhaled nitric oxide (FE(NO)) is thought to be a sensitive and reproducible method for assessing airway inflammation in asthmatics and the anti-inflammatory effects of drugs. A number of factors are known to contribute to intrapatient variation in FE(NO) which can confound interpretation. The aims of this study were to characterize the time-course of FE(NO), determine the effect of montelukast on the time-course of FE(NO), and evaluate the influence of the LTC(4) synthase A(-444)C polymorphism on montelukast-evoked changes in FE(NO). Following a 2-week run-in, 7 males and 5 females with asthma, 10-16 years old, received 5 or 10 mg of montelukast or an identical placebo at bedtime for 7 days in double-blind, crossover fashion, followed by a 7-day washout. FE(NO)was quantified every 30 min for 3 or 6 hr at baseline and on days 1, 2, 3, and 7 of treatment. A time-averaged value for FE(NO) was calculated (FE(NO)*), and % changes in FE(NO)* relative to baseline vs. time following placebo and montelukast were compared. The genotype of the A(-444)C polymorphism was determined by PCR and RFLP. FE(NO) varied markedly as a function of time in each patient. Time-averaged values of FE(NO) (FE(NO)*) during placebo and montelukast treatment were similar. Montelukast significantly reduced the slope of the % change in FE(NO)* vs. time curve in heterozygotes (n = 4), but not in A/A homozygotes (n = 8). These data suggest that heterozygotes respond better to montelukast compared to A/A homozygotes, at least with respect to changes in FE(NO). We conclude that assessment of inflammation or the anti-inflammatory effects of drugs in asthma based on single determinations of FE(NO) can be misleading. We further conclude that the A(-444)C polymorphism in the LTC(4) synthase gene probably contributes to interpatient variability in montelukast-evoked changes in FE(NO)* and warrants further study.

Acetates↗

Repeatability of sodium and chloride in exhaled breath condensates.

Exhaled breath condensate (EBC) has been proposed as a noninvasive tool to study airway inflammation. The reproducibility of breath condensates was recently questioned. We therefore measured sodium and chloride concentrations in EBC and assessed the repeatability of these measurements in healthy adults and children with airway disease. We investigated technical repeatability and within-day repeatability in five healthy adults, and compared these results with those of 10 asthmatic children and 9 children with cystic fibrosis (CF). We also assessed within-period repeatability in the healthy controls. We report that the variability of measurements was similar for within sample, within day, and between visits, for both normals and children with asthma and CF, and that the major source of variability of sodium and chloride measurements is restricted by the reproducibility of the measurement assay method used. The wide use of EBC is more likely to depend on the development of highly sensitive and reproducible assays, rather than further refinements of the collection technique.

Adult↗

Exhaled nitric oxide and asthma in young children.

Exhaled nitric oxide (eNO) has been used to diagnose asthma in adults and children using either the slow vital capacity method (SVCm) or, in younger children, the tidal breathing method (TBm). Adenosine 5'-monophosphate (AMP) challenge also has been found to be a sensitive and specific test for the diagnosis of asthma. In the present study, we used the AMP provocation concentration that caused wheezing (PCW) to confirm the diagnosis of asthma (PCW < or = 200 mg/mL). We studied 36 children (2-7 years) with mild intermittent asthma, 13 children (3-7 years) with moderate persistent asthma treated with inhaled steroids, 20 nonasthmatic children (2-7 years) with chronic cough and recurrent pneumonia, and 15 healthy children (4-6 years). Expired gas was collected in collection bags by the TBm, and eNO was measured. We evaluated the efficacy of eNO values in diagnosing asthma. The mean eNO level of the mild intermittent asthmatic children (5.6 +/- 0.4 ppb) not receiving inhaled corticosteroids was significantly higher (ANOVA P < 0.0001) than that of the moderate persistent asthmatics who were treated with inhaled steroids, the nonasthmatic children with chronic cough, and the group of healthy children (3.7 +/- 0.6 ppb, P < 0.05; 3.2 +/- 0.3 ppb, P < 0.001; 2.2 +/- 0.2 ppb, P < 0.001, respectively). The points of intersection for sensitivity and specificity curves of eNO to differentiate mild intermittent asthmatics from nonasthmatic children with chronic cough and from healthy children were 77% and 88% for eNO values of 3.8 ppb and 2.9 ppb, respectively. We conclude that eNO collected by the TBm can differentiate steroid-naive young children with intermittent asthma from healthy children, from nonasthmatic children with chronic cough, and from asthmatic children treated with inhaled steroids.

Analysis of Variance↗

Exhalation time effects on arterial and venous blood oxygen content and arterial PCO2 during high frequency jet ventilation of surfactant-depleted cats.

Since high frequency jet ventilation (HFJV) relies on lung mechanics for the passive removal of expiratory gas, one would predict that the time allowed for exhalation would have serious effects on cardiopulmonary function. To document these effects we lavaged the lungs of ten cats with 30 ml/kg of saline six times, then sampled arterial and venous blood while the animals were ventilated conventionally at 30 BPM and then with HFJV at 600 BPM, varying inspiratory/expiratory ratios (I/E) from 1:1 to 1:5. The animals breathed 100% O2 throughout the study, and the mean airway pressure was held constant for each animal when the I/E was varied during HFJV. Decreasing the I/E from 1:1 to 1:5 during HFJV resulted in an increase of arterial oxygen content (Cao2) from 11.3 +/- 1.2S E to 13.6 +/- 1.2 ml O2/100 ml blood (P less than 0.01), a decrease of PaCO2 from 43 +/- 6 to 27 +/- 4 mm Hg, and an increase of alveolar to arterial oxygen gradient from 351 +/- 49 to 377 +/- 49 mm Hg. The ratio of systemic blood flow to oxygen consumption (Q/VO2) was similar during conventional ventilation and with HFJV at I/E of 1:1 (18.9 +/- 3.7 vs 18.0 +/- 2.9) but decreased when I/E was reduced to 1:5 during HFJV (13.9 +/- 2.1). The ratio of the product of CaO2 and Q (systemic oxygen availability) to VO2 (SO2 T/VO2) remained unchanged during all modes of ventilation (1.75 +/- 0.15). The increase in CaO2 observed when I/E was reduced from 1:1 to 1:5 during HFJV was counterbalanced by a decrease in Q/VO2 such that SO2 T/VO2 remained relatively constant.

Animals↗

Effect of inhalation times on exhaled NO.

Recent work suggests that exhaled nitric oxide (eNO) can be divided into airway and alveolar components, using varying expiratory flow rates. It is known that higher values of eNO are found with lower expiratory flow rates, thought to be due to prolonged contact of the expirate with the bronchial epithelium. However, whether a prolonged inspiratory time could contaminate the alveolar signal with bronchial derived NO, by the same mechanism, is unknown. We aimed to study the effect, including repeatability, of inhalation time on eNO measurements performed at three different flow rates. We measured eNO in 21 children with asthma of different severity (no steroids, n = 2; inhaled corticosteroid (IC) dose up to 200 mcg/day, n = 3; IC up to 500 mcg/day, n = 3; IC >500 mcg/day, n = 6; IC >500 mcg/day and oral steroid, n = 7) and 24 normal adult controls at 50, 100, and 200 ml/s expiratory flow rate. The effect of either a rapid or a slow inhalation on measured eNO was studied at each flow rate. Furthermore, 12 asthmatic children and 12 adults had repeated measurements 1 hr apart. Repeatability within 1 day was assessed by calculating the single-determination standard deviation (SD) and 95% range. Our results showed that repeatability was equally good for the three expiratory flow rates, and inhalation time had no influence on the results. The inhalation maneuver does not influence eNO measurements using the variable expiratory flow technique.

Administration, Inhalation↗