Nitric Oxide In Exhaled Breath Research Articles

Proteomic analysis of sputum reveals novel biomarkers for various presentations of asthma

BackgroundIt is now recognized that asthma can present in different forms. Typically, asthma present with symptoms of wheeze, breathlessness and cough. Atypical forms of asthma such as cough variant asthma (CVA) or chest tightness variant asthma (CTVA) do not wheeze. We hypothesize that these different forms of asthma may have distinctive cellular and molecular features.Methods30 patients with typical or classical asthma (CA), 27 patients with CVA, 30 patients with CTVA, and 30 healthy control adults were enrolled in this prospective study. We measured serum IgE, lung function, sputum eosinophils, nitric oxide in exhaled breath (FeNO). We performed proteomic analysis of induced-sputum supernatants by mass spectrometry.ResultsThere were no significant differences in atopy and FEV1 among patients with CA, CVA, and CTVA. Serum IgE, sputum eosinophil percentages, FeNO, anxiety and depression scores were significantly increased in the three presentations of asthmatic patients as compared with healthy controls but there was no difference between the asthmatic groups. Comprehensive mass spectrometric analysis revealed more than a thousand proteins in the sputum from patients with CA, CVA, and CTVA, among which 23 secreted proteins were higher in patients than that in controls.ConclusionsPatients with CA, CVA, or CTVA share common clinical characteristics of eosinophilic airway inflammation. And more importantly, their sputum samples were composed with common factors with minor distinctions. These findings support the concept that these three different presentations of asthma have similar pathogenetic mechanism in terms of an enhanced Th2 associated with eosinophilia. In addition, this study identified a pool of novel biomarkers for diagnosis of asthma and to label its subtypes.Trial registrationhttp://www.chictr.org.cn (ChiCTR-OOC-15006221)

Bayesian estimation of physiological parameters governing a dynamic two-compartment model of exhaled nitric oxide.

The fractional concentration of nitric oxide in exhaled breath (fe NO) is a biomarker of airway inflammation with applications in clinical asthma management and environmental epidemiology. fe NO concentration depends on the expiratory flow rate. Standard fe NO is assessed at 50 mL/sec, but “extended NO analysis” uses fe NO measured at multiple different flow rates to estimate parameters quantifying proximal and distal sources of NO in the lower respiratory tract. Most approaches to modeling multiple flow fe NO assume the concentration of NO throughout the airway has achieved a “steady‐state.” In practice, this assumption demands that subjects maintain sustained flow rate exhalations, during which both fe NO and expiratory flow rate must remain constant, and the fe NO maneuver is summarized by the average fe NO concentration and average flow during a small interval. In this work, we drop the steady‐state assumption in the classic two‐compartment model. Instead, we have developed a new parameter estimation approach based on measuring and adjusting for a continuously varying flow rate over the entire fe NO maneuver. We have developed a Bayesian inference framework for the parameters of the partial differential equation underlying this model. Based on multiple flow fe NO data from the Southern California Children's Health Study, we use observed and simulated NO concentrations to demonstrate that our approach has reasonable computation time and is consistent with existing steady‐state approaches, while our inferences consistently offer greater precision than current methods.

Cytomegalovirus DNA is highly prevalent in the blood of patients with asthma and is associated with age and asthma traits.

Cytomegalovirus (CMV) IgG antibodies have been associated with inflammaging and immunosenescence. We aimed to assess the presence of CMV DNA in the blood of adult and elderly patients with bronchial asthma to establish potential association of CMV DNAemia with asthma and asthma characteristics. Eighty-five elderly asthmatics, 74 younger asthma patients, and 114 age-matched controls were recruited. The CMV DNA was detected using commercial artus assay in 10.7% of asthma patients, but was negative in all control individuals. The secondary assay identified CMV DNA in 41.5% of asthmatics and 13.3% of control subjects (P < .001). Presence of CMV DNA was associated with an increased risk of asthma and CMV DNA copy numbers correlated with some asthma traits, including respiratory parameters and exhaled breath nitric oxide. We conclude that CMV infection is associated with asthma and may contribute to the pathogenesis of asthmatic inflammation.

Determination of Normative Values of Fractional Nitric Oxide in Exhaled Breath (FeNO) in Healthy Indian Population

SESSION TITLE: Allergy - Asthma Slide SESSION TYPE: Original Investigation Slide PRESENTED ON: Wednesday, October 26, 2016 at 07:30 AM - 08:30 AM PURPOSE: Fractional nitric oxide in exhaled breath (FENO) is increased in diseases with airway inflammation, especially eosinophilic inflammation. To assess the utility of FENO in disease we need to know its normal value in healthy individuals. American thoracic society (ATS) has given upper limit of normal FENO values as 50 ppb but studies have shown variability in FENO values among different populations. There is no data available about FENO values in healthy Indians. METHODS: Study was conducted at a tertiary care centre in Delhi, India. Four hundred healthy non-smoking adults (age ≥18 years) with normal spirometry were enrolled. The subjects included medical students, hospital staff and patient relatives. Individuals with previous history of respiratory illness, allergic rhinits, atopy and family history of asthma were excluded. All underwent anthropometry and fractional exhaled nitric oxide measurement using hand held NIOX MINO - point of care device (electrochemical sensor based gas analyser). Population was divided into three arbitrary age groups - ≤ 30 years, 31-49 years and ≥50 years. Mann-Whitney U test was done for in-between group comparisons and Spearman’s correlation coefficient was assessed between FENO and various parameters. RESULTS: The mean age of sampled population was 39.6 ± 15.1 years and 263 (65.7%) were males. The median FENO value was 21 ppb (IQR 16-26, range 5-106) with upper and lower limit of normal being 41.5 ppb and 10 ppb respectively. The number of subjects in ≤ 30 years, 30-49 years and > 50 years age groups were 144, 123 and 133 respectively. The median (range, ULN) FENO value in subjects ≤ 30 years, 30-49 years and > 50 years age groups was 20 (6-106, 45) ppb, 22 (5-57, 41) ppb and 22 (7-103, 41) ppb respectively (p = 0.09). FENO values were significantly higher in males [median (IQR) 22 (18-28) ppb] compared to females [median (IQR) 18 (13-23) ppb] (p = 0.001). FENO correlated significantly with height, weight, FEV1 and FVC with correlation coefficients of 0.21 (p<0.001), 0.11 (p=0.02), 0.16 (p<0.001) and 0.18 (p<0.001) respectively. CONCLUSIONS: Upper limit of normal cut-off for FENO in healthy Indians was found to be 41.5 ppb and values above this in an individual will merit further investigation to assess for airway inflammatory states. CLINICAL IMPLICATIONS: Elucidation of normal FENO value in healthy Indian population will help in identifying individuals with airway inflammation among those presenting with respiratory symptoms. DISCLOSURE: The following authors have nothing to disclose: Saurabh Mittal, Anant Mohan, Randeep Guleria, Karan Madan, Vijay Hadda, Gopi Khilnani No Product/Research Disclosure Information

Study of the correlations between fractional exhaled nitric oxide in exhaled breath and atopic status, blood eosinophils, FCER2 mutation, and asthma control in Vietnamese children.

IntroductionFractional exhaled nitric oxide (FENO) is a biomarker of airway inflammation in asthma. The measurement of FENO is utilized to assist in the diagnosis and treatment of children with asthma, especially for those treated with inhaled corticosteroids.ObjectivesThe aims of this study were to evaluate the correlations between FENO and atopic status, blood eosinophil levels, FCER2 mutation, and asthma control in Vietnamese children.Subjects and methodsThis was a prospective and descriptive study approved by the local Ethical Board. All children with uncontrolled asthma, seen in the National Hospital of Pediatrics (Hanoi, Vietnam), were included. Exhaled breath FENO, blood eosinophils, skin prick test, total IgE, asthma control test (ACT), and FCER2 gene polymorphism were performed at inclusion. They were followed up at 3 months to evaluate clinical status, FENO levels, and ACT.ResultsForty-two children with uncontrolled asthma with a mean age of 10±3 years (6–16 years) were included. The male/female ratio was 2.5/1. The mean FENO levels were 26±25 ppb. FENO was significantly higher in patients with a positive skin prick test for respiratory allergens (P<0.05). FENO was significantly correlated with blood eosinophil levels (r=0.5217; P=0.0004). Five of the 32 subjects (15.6%) had a mutation of FCER2 gene (rs28364072 SNP). In this group, the levels of FENO were highest (37±10 ppb; P<0.05). The levels of FENO were significantly decreased after 3 months of treatment (17±8 ppb vs 26±25 ppb; P<0.05). Significant correlations between inhaled corticosteroid doses and FENO levels occurred at 1 and 3 months (r=0.415, P=0.007; r=0.396, P=0.010; respectively). There were no correlations between FENO levels, ACT, and daily use of salbutamol. After 3 months, asthma remained uncontrolled in 22.2% of children.ConclusionThe measurement of FENO levels is a useful and feasible tool to predict clinical, biological, and asthma control in Vietnamese children.

Reduced clot retraction rate and altered platelet energy production in patients with asthma

Objective: Asthma enhances the risk of pulmonary embolism. The mechanism of this phenomenon is unclear. Methods: We evaluated the kinetics of clot formation, clot retraction rate (CRR), clot volume at 40 min, the rate of lactate production (a marker of aerobic glycolysis in platelets in contracting clots), blood eosinophil count (EOS), nitric oxide in exhaled breath (FENO), and spirometry (FEV1) in 50 healthy controls and in 81 allergic asthmatics (41 subjects with steroid-naïve asthma and 40 with steroid-treated asthma). Results: Thromboelastometry revealed that only steroid-treated asthmatics had slightly activated coagulation. Compared with healthy controls, whole asthmatics demonstrated (p < 0.05) reduced CRR, higher clot volume at 40 minutes, higher FENO, decreased FEV1, elevated EOS, and augmented lactate production in retracting clots. Reduced CRR was observed also in the absence of native plasma. In whole study population (asthmatics and healthy controls), CRR positively correlated with spirometry (rS = 0.668, p = <0.001) and negatively with FENO (rS = −0.543; p < 0.001), EOS (rS = −0.367, p < 0.002), and lactate production (rS = −0.791; p < 0.001). However, in steroid-treated asthmatics, the CRR did not correlate with FENO and EOS. In all study patients lactate production negatively correlated with FEV1 and positively with FENO. Conclusion: Collectively, this data is consistent with the hypothesis that, in asthmatics, reactive nitrogen species produced in the lungs may reduce platelet contractility (and CRR) through the diminution of platelet energy production. CRR inhibition would predispose asthmatics to pulmonary embolism.

Evaluation of Acute Exogenous Hypoxia Impact on the Fraction of Exhaled Nitric Oxide in Healthy Males.

Exogenous hypoxia increases ventilation and contracts the pulmonary vessels. Whether those factors change the values of nitric oxide in exhaled air has not yet been evaluated. To examine the effect of exogenous normobaric hypoxia on the values of the fraction of nitric oxide in exhaled breath (FeNO). Subjects аnd Methods: Twenty healthy non-smoker males at mean age of 25.4 (SD = 3.7) were tested. The basal FeNO values were compared with those at 7 min. and 15 min. after introducing into the hypoxic environment (hypoxic tent), imitating atmospheric air with oxygen concentration corresponding to 3200 m above sea level. Exhaled breath temperature was measured at baseline and at 10-12 min. of the hypoxic exposition. Heart rate and oxygen saturation were registered by pulse-oximetry. All the subjects had FeNO values in the reference range. The mean baseline value was 14.0 ± 3.2 ppb, and in hypoxic conditions - 15.5 ± 3.8 ppb (7 min.) and 15.3 ± 3.6 ppb (15 min.), respectively, as the elevation is statistically significant (p = 0.011 and p = 0.008). The values of exhaled breath temperature were 33.79 ± 1.55°С and 33.87 ± 1.83°С (p = 0.70) at baseline and in hypoxic conditions, respectively. Baseline oxygen saturation in all subjects was higher than that, measured in hypoxia (96.93 ± 1.29% vs. 94.27 ± 2.53%; p < 0.001). Exogenous hypoxia leads to an increase of FeNO values, but does not affect the exhaled breath temperature.

Laboratory Safety of Capsaicin Inhalation in Healthy Younger and Older Populations Potential Template for Inhalation Research

Introduction: There are inconsistencies as to the effects of age on the human cough reflex. The investigation speaks to the Federal Drug Agency’s (FDA) trepidations for conducting inhalation experiments with non-approved medications/chemicals. The investigation addresses the accuracy of the mixing methodology and analysis of capsaicin stability during seven months of storage. Methods: There is stringent safety monitoring while conducting 12 serial (0.49-1000 uMol) single breath capsaicin inhalation challenge testing (CICT) on 20 younger and 20 older healthy research volunteers using pharmaceutical-grade capsaicin (i.e., Investigational New Drug protocol-IND 69,642). The research design and subject safety measures are recommended by the Food & Drug Agency (FDA) and approved by the Institutional Review Board (IRB). Mixing of inhaled capsaicin solutions are by a Registered Pharmacist and concentration verifications are by high performance liquid chromatography (HPLC). Potency of stored capsaicin (i.e., refrigerated and shielded from UV light) is examined over 7 months. Results: There are neither adverse reactions nor statistically significant difference in capsaicin cough parameters for older and younger volunteers at any dose of capsaicin. Physiologic monitoring by spirometry, impulse oscillometry, exhaled breath nitric oxide, electrocardiography, blood pressure, pulse and oxygen saturation measurements do not change at any dose. There are differences between the concentrations of capsaicin solutions mixed by a Registered Pharmacist and actual capsaicin determination by HPLC. The differences in capsaicin concentrations are 28.1% lower for 0.49 uMol compared to a 2.2% lesser concentration for 1000 uMol solution. During storage, capsaicin remains stable for 3-months but substantially falls by six (p<0.03) and seven (p<0.004) months, especially for the lower concentrations. Conclusion: The results of this investigation embrace the safety of serial Inhalations of dilute pharmacologic grade capsaicin aerosol among older and younger normal volunteers. Incorporating rigorous FDA recommendations, inclusion of IRB oversight and monitoring by spirometry, impulse oscillometry, exhaled breath nitric oxide, electrocardiography, blood pressure, pulse and oxygen saturation measurements document inhalation safety at any dilute capsaicin aerosol dose. However, there are lower capsaicin values determined by HPLC compared to expected capsaicin concentrations when mixed by a Registered Pharmacist or after six months of storage. The investigation champions the opinion that inhalation studies involving FDA non-approved drugs or chemicals/ medications can be safely conducted when following the appropriate safety procedures such as in this research protocol.

Clinical Effects, Exhaled Breath Condensate pH and Exhaled Nitric Oxide in Humans After Ethyl Acrylate Exposure.

Ethyl acrylate is an irritant known to affect the upper airways and eyes. An increase of the eye blink frequency in humans was observed during exposure to 5ppm. Studies on the lower airways are scant and our study objective was the evaluation of pH in exhaled breath condensate (EBC-pH) and nitric oxide in exhaled breath (FeNO) as markers of inflammation. Sixteen healthy volunteers were exposed for 4h to ethyl acrylate at a concentration of 5ppm and to sham (0.05ppm) in an exposure laboratory. Clinical irritation symptoms, EBC-pH (at a pCO2 of 5.33kPa) and FeNO were assessed before and after exposure. Differences after ethyl acrylate exposure were adjusted for those after sham exposure. 5ppm ethyl acrylate induced clinical signs of local irritation in the nose and eyes, but not in lower airways. Exposure produced a subtle, but statistically significant, decrease in breathing frequency (1breath/min; p = 0.017) and a lower EBC-pH (by 0.045 units; p = 0.037). Concerning FeNO, we did not observe significant changes compared to sham exposure. We conclude that local effects induced by 5ppm ethyl acrylate consist of sensory irritation of eyes and nose. In addition, acute ethyl acrylate exposure to 5ppm resulted in a net decrease of EBC-pH. Whether that can be interpreted in terms of additional lower airway irritation or already inflammatory alterations set in needs further investigations.

Exhaled NO: Determinants and Clinical Application in Children With Allergic Airway Disease.

Nitric oxide (NO) is endogenously released in the airways, and the fractional concentration of NO in exhaled breath (FeNO) is now recognized as a surrogate marker of eosinophilic airway inflammation that can be measured using a noninvasive technique suitable for young children. Although FeNO levels are affected by several factors, the most important clinical determinants of increased FeNO levels are atopy, asthma, and allergic rhinitis. In addition, air pollution is an environmental determinant of FeNO that may contribute to the high prevalence of allergic disease. In this review, we discuss the mechanism for airway NO production, methods for measuring FeNO, and determinants of FeNO in children, including host and environmental factors such as air pollution. We also discuss the clinical utility of FeNO in children with asthma and allergic rhinitis and further useful directions using FeNO measurement.

Development of a respiratory sensitization/elicitation protocol of toluene diisocyanate (TDI) in Brown Norway rats to derive an elicitation-based occupational exposure level

Toluene diisocyanate (TDI), a known human asthmagen, was investigated in skin-sensitized Brown Norway rats for its concentration×time (C×t)-response relationship on elicitation-based endpoints. The major goal of study was to determine the elicitation inhalation threshold dose in sensitized, re-challenged Brown Norway rats, including the associated variables affecting the dosimetry of inhaled TDI-vapor in rats and as to how these differences can be translated to humans. Attempts were made to duplicate at least some traits of human asthma by using skin-sensitized rats which were subjected to single or multiple inhalation-escalation challenge exposures. Two types of dose-escalation protocols were used to determine the elicitation-threshold C×t; one used a variable C (Cvar) and constant t (tconst), the other a constant C (Cconst) and variable t (tvar). The selection of the “minimal irritant" C was based an ancillary pre-studies. Neutrophilic granulocytes (PMNs) in bronchoalveolar lavage fluid (BAL) were considered as the endpoint of choice to integrate the allergic pulmonary inflammation. These were supplemented by physiological measurements characterizing nocturnal asthma-like responses and increased nitric oxide in exhaled breath (eNO). The Cconst×tvar regimen yielded the most conclusive dose–response relationship as long C was high enough to overcome the scrubbing capacity of the upper airways. Based on ancillary pre-studies in naïve rats, the related human-equivalent respiratory tract irritant threshold concentration was estimated to be 0.09ppm. The respective 8-h time-adjusted asthma-related human-equivalent threshold C×t-product (dose), in ‘asthmatic’ rats, was estimated to be 0.003ppm. Both thresholds are in agreement of the current ACGIH TLV® of TDI and published human evidence. In summary, the findings from this animal model suggest that TDI-induced respiratory allergy is likely to be contingent on two interlinked, sequentially occurring mechanisms: first, dermal sensitizing encounters high enough to cause systemic sensitization. Second, when followed by inhalation exposure(s) high enough to initiate and amplify an allergic airway inflammation, then a progression into asthma may occur. This bioassay requires an in-depth knowledge on respiratory tract dosimetry and irritation of the involved test substance to clearly understand the dosimetry causing C- and/or C×t-dependent respiratory tract irritation and eventually asthma.

Different Patterns in Changes of Exhaled Breath Condensate pH and Exhaled Nitric Oxide After Ozone Exposure.

Study objective was the evaluation of pH in exhaled breath condensate (EBC-pH) and nitric oxide in exhaled breath (FeNO) as biomarkers of ozone induced inflammation. We recently demonstrated that an ozone exposure of 240 ppb is sufficient to reduce lung function indices. We enrolled ten healthy subjects exposed in an intermittent exercise protocol to ozone concentrations of 240 ppb and 40 ppb (sham exposure). EBC-pH and FeNO were assessed before (pre), immediately post (post), and 16 h after exposure (16 h). Findings are that compared to baseline, EBC-pH was significantly higher immediately after sham and ozone exposures, but not 16 h later. There was a negative net change in EBC-pH after adjusting for effects after sham exposure (net-ΔpHpost -0.38%, net-ΔpH16h -0.23%). Concerning FeNO, we observed no changes of values after sham exposure compared to baseline, but measured a significant lower net response at the end of exposure (net-ΔFeNOpost -17.5%) which was transient within 16 h (net-ΔFeNO16h -9.4%). We conclude that exercise known to enhance EBC-pH may compensate for EBC acidification associated with inflammation resulting in diminished change of this biomarker. Ozone imposes an oxidative burden and reactions between reactive oxygen species and NO might be an explanation for reduced FeNO levels.

P87 Is fraction of exhaled nitric oxide (FeNO) in asymptomatic older teenagers related to preschool wheeze and chronic cough?

BackgroundMany children with preschool wheeze and chronic cough become asymptomatic in later childhood, only for asthma to be diagnosed in adulthood (1). A previous study found in individuals with apparently...

Relations between isoprene and nitric oxide in exhaled breath and the potential influence of outdoor ozone: a pilot study

The role of endogenous isoprene in the human body, if any, is unclear because previous research is inconsistent and mechanistic evidence for the biologic function of isoprene is lacking. Given previous evidence that exhaled isoprene is elevated in systemic inflammatory states, we hypothesized that exhaled isoprene would be positively associated with a breath biomarker of airway inflammation, the fractional concentration of exhaled nitric oxide (FENO). We examined relationships of exhaled breath isoprene with FENO and with outdoor ozone given that ozone chemically reacts with isoprene and has been positively associated with FENO in past studies. Sixteen elderly subjects were followed with ≤12 weekly exhaled hydrocarbon and FENO collections at the subjects’ retirement community. Outdoor ozone concentrations were measured continuously on site. Mixed-effects regression analyses tested relations of FENO with isoprene, and FENO and isoprene with ozone, adjusted for temperature. We found FENO was inversely associated with isoprene, and this was not confounded by ozone. Isoprene was inversely related to ozone. FENO was positively related to ozone and this relation was not confounded by isoprene. In contrast to hypothesized relations, we conclude that exhaled isoprene is inversely associated with FENO as well as outdoor ozone, which suggests possible protective ozone-scavenging functions of endogenous isoprene. Findings may indicate chemical reactions of isoprene oxidation by ozone and by hydroxyl radicals in the presence of O2 that is dependent on NO concentration. These preliminary results need to be confirmed in additional studies of human subjects, particularly as they apply to FENO monitoring in asthma.

The value of FeNO measurement in childhood asthma: uncertainties and perspectives

Asthma is considered an heterogeneous disease, requiring multiple biomarkers for diagnosis and management. Fractional exhaled nitric oxide in exhaled breath (FeNO) was the first useful non-invasive marker of airway inflammation in asthma and still is the most widely used. The non-invasive nature and the relatively easy use of FeNO technique make it an interesting tool to monitor airway inflammation and rationalize corticosteroid therapy in asthmatic patients, together with the traditional clinical tools (history, physical examination and lung function tests), even if some controversies have been published regarding the use of FeNO to support the management of asthma in children. The problem of multiple confounding factors and overlap between healthy and asthmatic populations preclude the routine application of FeNO reference values in clinical practice and suggest that it would be better to consider an individual “best”, taking into account the context in which the measurement is obtained and the clinical history of the patient. Besides, there is still disagreement about the role of FeNO as a marker of asthma control, due to the complexity of balance among the different items involved in its determination and the lack of homogeneity in the population groups studied in the few studies conducted so far. Heterogeneity of problematic severe asthma greatly limits utility of FeNO alone as a biomarker of inflammation to optimize the disease management on an individual basis. None of the studies conducted so far demonstrated that the use of FeNO was better than current asthma guidelines in controlling asthma exacerbations. In summary, there is a large variation in FeNO levels between individuals, which may reflect the natural heterogeneity in baseline epithelial nitric oxide synthase activity and/or the contribution of other noneosinophilic factors to epithelial nitric oxide synthase activity. FeNO is a promising biomarker, but at present some limits are highlighted. We would recommend that further research can be carried out by organizing studies aimed to obtain reliable reference values of FeNO and in order to better interpret FeNO measurements in clinical settings, taking also into account the influence of genetic and environmental factors.

Nitric oxide in the breath of bottlenose dolphins: Effects of breath hold duration, feeding, and lung disease

AbstractBreath analysis, including measurement of nitric oxide (NO), is a noninvasive diagnostic tool that may help evaluate cetacean health. This is the first report on the effects of breath hold duration, feeding, and lung disease on NO in dolphin exhaled breath. Three healthy dolphins were trained to hold their breath for 30, 60, 90, and 120 s and then exhale into an underwater funnel. Exhaled NO values from 157 breath samples were compared among three healthy dolphins by breath hold time and after fasting and feeding. Exhaled NO values were also measured in two dolphins with pulmonary disease. NO in dolphin breath was higher compared to ambient air; healthy dolphins had higher NO concentrations in their breath after feeding compared to after overnight fasting; and there were no significant differences in exhaled NO levels by breath hold duration. A dolphin with Mycoplasma‐associated pneumonia and chronic gastrointestinal disease had higher postprandial exhaled NO levels compared to healthy controls. This study demonstrates, contrary to previous publications, that dolphins exhale NO. Given the high standard deviations present in exhaled breath NO values, future studies are needed to further standardize collection methods or identify more reliable samples (e.g., blood).

When are biomarkers useful in the management of airway diseases?

Biomarkers are characteristics that are objectively measured and evaluated as indicators of biological or pathogenic processes, or responses to therapeutic interventions, and may provide information on the prognosis or progression of the disease and response to treatment. They are likely to be helpful in the management of airway diseases because of the heterogeneity of their pathobiology. Most biomarkers have been developed and evaluated to assess the airway inflammation (or bronchitis) associated with airway diseases. These include quantitative cell counts in sputum, fraction of nitric oxide in exhaled breath, and various metabolites in exhaled breath. This review provides a brief description of these biomarkers with a particular emphasis on how eosinophil and neutrophil counts in sputum could be used to manage airway diseases such as asthma, chronic obstructive pulmonary disease, and chronic cough.

Methods of NO detection in exhaled breath

There is still an unexplored potential for exhaled nitric oxide (NO) in many clinical applications. This study presents an overview of the currently available methods for monitoring NO in exhaled breath and the use of the modelling of NO production and transport in the lung in clinical practice. Three technologies are described, namely chemiluminescence, electrochemical sensing and laser-based detection with their advantages and limitations. Comparisons are made in terms of sensitivity, time response, size, costs and suitability for clinical purposes. The importance of the flow rate for NO sampling is discussed from the perspective of the recent recommendations for standardized procedures for online and offline NO measurement. The measurement of NO at one flow rate, such as 50 ml s−1, can neither determine the alveolar site/peripheral contribution nor quantify the difference in NO diffusion from the airways walls. The use of NO modelling (linear or non-linear approach) can solve this problem and provide useful information about the source of NO. This is of great value in diagnostic procedures of respiratory diseases and in treatment with anti-inflammatory drugs.

The value of FeNO measurement in childhood asthma: uncertainties and perspectives.

Asthma is considered an heterogeneous disease, requiring multiple biomarkers for diagnosis and management. Fractional exhaled nitric oxide in exhaled breath (FeNO) was the first useful non-invasive marker of airway inflammation in asthma and still is the most widely used. The non-invasive nature and the relatively easy use of FeNO technique make it an interesting tool to monitor airway inflammation and rationalize corticosteroid therapy in asthmatic patients, together with the traditional clinical tools (history, physical examination and lung function tests), even if some controversies have been published regarding the use of FeNO to support the management of asthma in children. The problem of multiple confounding factors and overlap between healthy and asthmatic populations preclude the routine application of FeNO reference values in clinical practice and suggest that it would be better to consider an individual “best”, taking into account the context in which the measurement is obtained and the clinical history of the patient. Besides, there is still disagreement about the role of FeNO as a marker of asthma control, due to the complexity of balance among the different items involved in its determination and the lack of homogeneity in the population groups studied in the few studies conducted so far. Heterogeneity of problematic severe asthma greatly limits utility of FeNO alone as a biomarker of inflammation to optimize the disease management on an individual basis. None of the studies conducted so far demonstrated that the use of FeNO was better than current asthma guidelines in controlling asthma exacerbations. In summary, there is a large variation in FeNO levels between individuals, which may reflect the natural heterogeneity in baseline epithelial nitric oxide synthase activity and/or the contribution of other noneosinophilic factors to epithelial nitric oxide synthase activity. FeNO is a promising biomarker, but at present some limits are highlighted. We would recommend that further research can be carried out by organizing studies aimed to obtain reliable reference values of FeNO and in order to better interpret FeNO measurements in clinical settings, taking also into account the influence of genetic and environmental factors.

Exhaled NO is a poor marker of asthma control in children with a reported use of asthma medication: a pharmacy‐based study

A high fraction of nitric oxide in exhaled breath (FeNO) has been suggested to be a marker of ongoing airway inflammation and poorly controlled disease in asthma. The usefulness of FeNO to monitor asthma control is still debated today. To assess the validity of FeNO as a marker of asthma control in children with reported use of asthma medication. Fraction of nitric oxide in exhaled breath was measured in 601 children (aged 4-12 yr) with reported use of asthma medication in the past 6 months and in 63 healthy non-asthmatic children (aged 5-12). Asthma control was assessed by the Asthma Control Questionnaire (ACQ). A receiver-operator characteristics (ROC) curve was generated to assess the accuracy of FeNO as a marker for asthma control. Logistic regression analysis was used to study whether clinical, healthcare, medication, and environmental factors are associated with high FeNO levels (>25 ppb). Fraction of nitric oxide in exhaled breath had a poor accuracy to discriminate well-controlled from not well-controlled asthma [area under the ROC curve: 0.56 (95% CI: 0.52-0.61, p = 0.008)]. In addition, high FeNO (>25 ppb) was associated with lower medication adherence rates (OR: 0.4; 95% CI 0.3-0.6), fewer antibiotic courses in the past year (OR: 0.6; 95% CI: 0.4-0.9), fewer leukotriene antagonists use in the past year (OR: 0.4; 95% CI: 0.2-0.9), and fewer visits to a (pulmonary) pediatrician (OR: 0.6; 95% CI: 0.4-0.9). Children living in a non-urban environment had more often high FeNO levels (OR: 1.7; 95% CI: 1.1-2.6). High FeNO is a poor marker of asthma control in children with reported use of asthma medication. Various other factors, including medication adherence and medication use, are associated with increased FeNO levels.