Markers In Exhaled Breath Condensate Research Articles

Inflammatory markers in exhaled breath condensate in nonasthmatic children with food allergy

Allergy is a systemic inflammation; therefore, although the allergic symptom may be seen in a specific organ system, the effects of this inflammation may be seen in other organs. interleukin (IL) IL4, IL5 and IL13 are the major Th2 cytokines and e-cadherin is an epithelial barrier protein. The objective of this research was to assess indicators of inflammation specific to Th2 responses and proteins related to the protective barrier of the airway’s inner lining. These assessments were conducted using exhaled breath condensate (EBC), which provides insights into peripheral airway conditions of children suffering from food allergies. The study had 24 patients with food allergy and 24 control individuals younger than three years of age with no history of food reaction. The diagnosis of food allergy was based on food allergen-specific IgE and skin prick test positivity in our clinic and oral food testing in selected cases. EBC samples were obtained by Ecoscreen (Jaegar, Hoechberg, Germany). IL4, IL5, IL13 and E-cadherin levels were measured in these samples by enzyme linked immunoassay. The group of children with food allergies, consisting mainly of 14 girls, had a median age of 16 months, whereas the control group, which included 11 girls, had a median age of 15 months (p = 0.89). Comparing the two groups, children with food allergies exhibited notably lower levels of IL-13 in the EBC compared to the control group (median values of 59.14 and 76.36, respectively, p = 0.02). Conversely, the concentration of IL-4 in the EBC was significantly higher in children with food allergies (median values of 1.94 and 1.29, respectively, p = 0.003). However, the levels of IL-5 and e-cadherin showed no significant differences between the two groups (with p-values of 0.74 and 0.09, respectively) as shown in table . High level of IL-4 despite the low level of IL-13 in the EBC of children having food allergy may be indicative of an early inflammatory phase that is not yet in the effector phase. Studies about the evolution of this process later in life are needed to assess the role of airway inflammation in children with food allergy who develop asthma.

Exhaled Breath Condensate and Respiratory Sequelae in Children Post-COVID-19

Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes an acute respiratory illness. A substantial proportion of adults experience persistent symptoms. There is a paucity of data on respiratory sequelae in children. Exhaled breath condensate (EBC) is a non-invasive tool used to assess airway inflammation. Objectives: This study aimed to evaluate EBC parameters, respiratory, mental and physical ability among children post COVID-19 infection. Methods: Observational study of confirmed SARS-CoV-2 infection cases among children, aged 5–18 years, evaluated once, 1–6 months post positive SARS-CoV-2 PCR testing. All subjects performed spirometry, 6-min walk test (6MWT), EBC (pH, interleukin-6), and completed medical history questionnaires, Depression, Anxiety, and Stress Scale (DASS-21), and physical activity scores. Severity of COVID-19 disease was classified according to WHO criteria. Results: Fifty-eight children were included and classified asymptomatic (n = 14), mild (n = 37), and moderate (n = 7) disease. The asymptomatic group included younger patients compared to the mild and moderate groups (8.9 ± 2.5y vs. 12.3 ± 3.6y and 14.6 ± 2.5y, respectively, p = 0.001), as well as lower DASS-21 total scores (3.4 ± 4 vs. 8.7 ± 9.4 and 8.7 ± 0.6 respectively, p = 0.056), with higher scores in proximity to positive PCR (p = 0.011). No differences were found between the 3 groups regarding EBC, 6MWT, spirometry, body mass index percentile, and activity scores. Conclusions: COVID-19 is an asymptomatic-mild disease in most young healthy children, with gradually diminishing emotional symptoms. Children without prolonged respiratory symptoms revealed no significant pulmonary sequelae as evaluated by EBC markers, spirometry, 6MWT, and activity scores. Larger studies are required to assess long-term pediatric consequences of post SARS‐CoV‐2 infection, to assess the need for pulmonology surveillance.

Diet Quality and Exhaled Breath Condensate Markers in a Sample of School-Aged Children.

Exhaled breath condensate (EBC) analysis is a recently developed, non-invasive method used to identify and quantify biomarkers, mainly those coming from the lower respiratory tract. It seems that diet can influence the airway's inflammation and change the exhaled breath composition. This study aimed to assess the association between diet quality intake and markers in EBC among school-aged children. A cross-sectional analysis included 150 children (48.3% females, aged 7-12 years, mean age: 8.7 ± 0.8 years) from 20 schools across Porto, Portugal. We assessed diet quality through the Healthy Eating Index (HEI)-2015, which was estimated based on a single 24 h food recall questionnaire. EBC samples were collected, and we assessed their ionic content (Na+, K+) and conductivity. The association between diet quality and Na+, K+, Na+/K+ ratio and conductivity was estimated using logistic regression models adjusted for potential confounders. After adjustment, a higher quality diet score increases the odds of higher conductivity values of the EBC (aOR = 1.04, 95%CI 1.00; 1.08). Our findings suggest that a higher diet quality in school-aged children is associated with higher conductivity levels of the EBC.

Inflammatory markers in exhaled breath condensate in bronchial asthma

Chronic respiratory diseases are among the most common non- infection diseases. In particular, it is bronchial asthma (BA), characterized by bronchial hyperreactivity and varying degrees of airway obstruction that is the cause of morbidity and mortality. The methods available for the information about the presence of inflammation in the airways, such as bronchoscopy and bronchial biopsy to be obtained have currently been invasive and difficult in everyday clinical practice, especially for children and seriously ill patients. In this regard, recently there has been an increase in the development of non-invasive methods for diagnosing the respiratory system, being comfortable and painless for trial subjects, especially children, also providing the inflammatory process control in the lungs, the severity assessment and monitoring the treatment process. The exhaled breath condensate (EBC) is of great attention, which is a source of various biomolecules, including nitric oxide (NO), leukotrienes, 8-isoprostane, prostaglandins, etc., being locally or systemically associated with disease processes in the body. Of particular interest is the presence of cytokines in EBC, namely the specific proteins produced by various cells of the body that play a key role in inflammatory processes in AD and provide cell communication (cytokine network). Thereby, it becomes possible for the severity and control level of childhood bronchial asthma using only the EBC analysis to be assessed. In addition, the non-invasiveness of this method allows it to be reused for monitoring lung diseases of even the smallest patients, including infants. Thus, the field of metabolite analysis in EBC has been developing and, in the near future, the given method is likely to be the most common for diagnosing the respiratory system diseases in both children and adults.

Markers of oxidative stress after three days of nanoTiO<sub>2</sub> sunscreen use in humans: a pilot study

Recent experimental studies point to a high reactivity of nanoparticles and the potential of sunscreens to penetrate the skin. We measured 20 markers of oxidative stress and inflammation to find out whether skin exposure to nanoTiO2 sunscreen may elevate the level of the markers in exhaled breath condensate (EBC) and urine of exposed subjects, as was suggested by our earlier study. Six volunteers (3 males and 3 females), with a mean age of 48.0 ± 6.7 years, used commercial sunscreen for three days continuously. The first samples were collected before the test. The second samples were collected on day 4, before the sunscreen was washed off, and the third samples on day 11. The following biomarkers were measured: malondialdehyde, 4-hydroxy-trans-hexenal, 4-hydroxy-trans-nonenal, aldehydes C6-C12, 8-isoProstaglandin F2α, o-tyrosine, 3-chlorotyrosine, 3-nitrotyrosine, 8-hydroxy-2-deoxyguanosine, 8-hydroxyguanosine, 5-hydroxymethyl uracil, and leukotrienes B4, C4, D4, and E4, using liquid chromatography-electrospray ionisation-tandem mass spectrometry. In the urine, 4-hydroxy-trans-hexenal was significantly higher in post-exposure sample 2, and the same trend was seen in all urinary markers. In EBC, no difference was seen between the mean values of 20 post-test markers as compared with pre-test samples. This study suggests potential side effects of the sunscreen - borderline elevation of markers of oxidative stress/inflammation - which may relate to the absorption of the nanoTiO2, and the non-significant difference may be explained by the small number of subjects. The effect was not seen in EBC, where nanoTiO2 was not found. A larger study is needed, as according to our previous study, the beneficial effect of the sunscreen to suppress oxidative stress caused by UV radiation may be questioned.

The relationship between oxidative stress markers in exhaled breath condensate and respiratory problems in patients with repaired esophageal atresia

AimTo evaluate the relationship between respiratory problems and oxidative stress markers in exhaled breath condensate (EBC) of patients with esophageal atresia (EA). MethodsEA cases with respiratory problems were evaluated retrospectively for age, gender, the type of atresia, surgical treatment, outcome and respiratory symptoms. The results of gastroesophageal reflux (GER) treatment including the use of proton pump inhibitor (PPI) and fundoplication were also documented. EBC samples of 500–1000 μl were obtained by Ecoscreen machine in all cases. The levels of Glutathione (Glut), 8-isoprostane (8-iso), cysteinyl-leukotriene (Cys-LT) were measured with ELISA. Results were compared with healthy control subjects (CG, n = 26) and the relationship between oxidative stress markers and respiratory symptoms was evaluated. The results of GER treatment and oxidative stress markers in EBC were also correlated. ResultsTwenty-nine patients with a mean age of 8.8 years (3–14 years) were included. The male/female ratio was 16:13. The EA presented with distal fistula in 27 cases. While no fistula was observed in 1 case, both proximal and distal fistulae were present in another single case. Associated anomalies, most of which were cardiovascular anomalies, were observed in 65.5% (n = 19) of cases. The median Glut level was 1.03 mM/ml (0.93–1.15), iso-8 was 38.8 pg/mL (32.03–76.2) and Cys-LT was 0.44 pg/mL (20.17–61.3) in patients with EA. The median levels of oxidative markers in CG were 1.23 mM/mL (1.13–1.36), 66.3 pg/mL (33.5–106.7), and 56.9 pg/mL (27.4–80.1), respectively. Glut levels were significantly lower in EA cases compared to CG (p = 0.01). There was no significant difference between the groups regarding 8-iso and CYS-LT levels (p = 0.9, p = 1.0). Cys-LT levels were significantly lower in patients with PPI treatment [21.7 pg/mL (18.6–48.1)], when compared to patients without PPI treatment [41.1 pg/mL (22.5–83.1)] (p = 0.04) and healthy subjects [56.9 pg/mL (27.4–80.1)] (p = 0.017). The 8-iso levels were significantly decreased in cases with fundoplication compared to the patients without fundoplication (p = 0.02). ConclusionGlut – an antioxidant agent – levels were significantly lower in EBC of EA cases. The decrease in Cys-LT levels in cases with PPI treatment and in 8-iso levels in patients with fundoplication suggests that the oxidative damage in EBC of EA cases may be correlated with GER and its management. Type of studyCase control study Level of evidenceLevel III

Exhaled Breath Condensate in Childhood Asthma: A Review and Current Perspective.

Exhaled breath condensate (EBC) was introduced more than two decades ago as a novel, non-invasive tool to assess airway inflammation. This review summarizes the latest literature on the various markers in EBC to predict asthma in children. Despite many recommendations and two comprehensive Task Force reports, there is still large heterogeneity in published data. The biggest issue remains a lack of standardization regarding EBC collection, preservation, processing, and analysis. As a result, published studies show mixed or conflicting results, questioning the reproducibility of findings. A joint, multicenter research study is urgently needed to address the necessary methodological standardization.

Exhaled breath condensate biomarkers reflect systemic changes in patients with chronic dioxin intoxication

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is highly toxic and affects the cardiovascular system, brain, and skin by AhR-dependent and other mechanisms, as well as causing metabolic impairments and cancer. The involvement of the respiratory system has not yet been studied. TCDD in the blood was measured and biomarkers of oxidative stress and inflammation were analysed in 2016 in the exhaled breath condensate (EBC) of the last eight male survivors (mean age 72.4 ± 1.3 years) from 80 workers intoxicated with TCDD during the production of herbicides from 1965 to 1968. The results were compared with their findings in 2010 to evaluate a trend. Malondialdehyde, 4-hydroxy-trans-nonenale, and 8-isoprostaglandin F2α (8-isoprostane), in addition to markers of the oxidation of nucleic acids and proteins 8-hydroxy-2-deoxyguanosine, 8-hydroxyguanosine, 5-(hydroxymethyl)uracil, o-tyrosine, and 3-nitrotyrosine, as well as markers of inflammation leukotrienes and anti-inflammatory lipoxins, were analysed in EBC by liquid chromatography–electrospray ionisation–tandem mass spectrometry. In addition, the patients underwent chest X-ray, spirometry and fractional exhaled nitric oxide (FeNO) examinations. The control group included 7 men (66 ± 16 years) with comparable lifestyle factors. The median plasma TCDD level lowered from 155 (28–553) ng/kg fat in 2010 to 112 (46–390) ng/kg fat in 2016, i.e., 50 years after exposure. The mean TCDD body deposit was 5.0 ± 3.7 µg. Serum TCDD level in the pooled sample of the controls was 12 ng/kg fat. All markers of oxidative stress, LTB4 and LTC4, remained overexpressed in patients and anti-inflammatory lipoxins were under-expressed compared to controls (all p < 0.01). The mean FeNO and spirometry results were within the reference values. Borderline X-ray findings and combined lung function impairments were seen in the patients with the lower TCDD plasma levels. Differences in the expression of the biomolecular markers in EBC as compared to controls were not associated with lung impairments and the respiratory parameters measured. Therefore, these EBC markers can be used to evaluate systemic oxidative stress and inflammation in tissues and the endovascular, atherosclerotic, neurotoxic, and metabolic effects of TCDD.

Markers of lipid oxidative damage among office workers exposed intermittently to air pollutants including nanoTiO2 particles.

Nanoscale titanium dioxide (nanoTiO2) is a commercially important nanomaterial used in numerous applications. Experimental studies with nanotitania have documented lung injury and inflammation, oxidative stress, and genotoxicity. Production workers in TiO2 manufacturing with a high proportion of nanoparticles and a mixture of other air pollutants, such as gases and organic aerosols, had increased markers of oxidative stress, including DNA and protein damage, as well as lipid peroxidation in their exhaled breath condensate (EBC) compared to unexposed controls. Office workers were observed to get intermittent exposures to nanoTiO2 during their process monitoring. The aim of this study was to investigate the impact of such short-term exposures on the markers of health effects in office workers relative to production workers from the same factory. Twenty-two office employees were examined. They were occupationally exposed to (nano)TiO2 aerosol during their daily visits of the production area for an average of 14±9 min/day. Median particle number concentration in office workers while in the production area was 2.32×104/cm3. About 80% of the particles were <100 nm in diameter. A panel of biomarkers of lipid oxidation, specifically malondialdehyde (MDA), 4-hydroxy-trans-hexenal (HHE), 4-hydroxy-trans-nonenal (HNE), 8-isoprostaglandin F2α (8-isoprostane), and aldehydes C6-C12, were studied in the EBC and urine of office workers and 14 unexposed controls. Nine markers of lipid oxidation were elevated in the EBC of office employees relative to controls (p<0.05); only 8-isoprostane and C11 were not increased. Significant association was found in the multivariate analysis between their employment in the TiO2 production plant and EBC markers of lipid oxidation. No association was seen with age, lifestyle factors, or environmental air contamination. The EBC markers in office employees reached about 50% of the levels measured in production workers, and the difference between production workers and office employees was highly significant (p<0.001). None of these biomarkers were elevated in urine. The approach presented here seems to be very sensitive and useful for non-invasive monitoring of employees exposed to air pollutants, including gases, organic aerosols, and nanoTiO2, and may prove useful for routine biomonitoring purposes. Among them, aldehydes C6, C8, C9, and C10 appear to be the most sensitive markers of lipid oxidation in similar occupational cohorts. One major challenge with sensitive biomonitoring techniques, however, is their non-specificity and difficulty in interpreting the meaning of their physiological values in the context of chronic disease development and damage-repair kinetics.

Serum tryptase level and inflammatory markers in exhaled breath condensate of children with exercise-induced symptoms.

The pathogenesis of exercise-induced bronchoconstriction (EIB) is poorly understood. To evaluate the biomarkers concentration in exhaled breath condensate (EBC) in schoolchildren with postexercise symptoms. We also evaluated changes in fractional exhaled nitric oxide (FeNO) value and the serum tryptase level after exercise. One hundred children with postexercise symptoms were included. Methacholine challenge testing (MCT) was performed at visit 2, and exercise challenge testing (ECT) was performed at visit 3. Before and after ECT serum tryptase levels and FeNO values were measured. EBC was collected after ECT from 10 randomly selected children from each group. The children were assigned to the following groups: ECT(+) MCT(+), ECT(+) MCT(-), ECT(-) MCT(+), ECT(-) MC(-). We measured the following molecules: eotaxin, interleukin (IL) 8, IL-1ra, IL-1 beta, IL-6, IL-1 alpha, IL-12(p40), IL-5, granulocyte-macrophage colony-stimulating factor, IL-7, IL-15, IL-4, IL-2, IL-10, tumor necrosis factor alpha, interferon gamma, IL-13, tumor necrosis factor beta, monocyte chemoattractant protein-1, IL-17A, macrophage inflammatory proteins-1 alpha, macrophage inflammatory proteins-1 beta, IL-12(p70), and regulated on activation, normal T-cell expressed and secreted by using a multiplex immunoassay. Prostaglandin E2 (PGE2), leukotriene B4, and cysteinyl leukotriene were analyzed by using separate enzyme-linked immunosorbent assay kits. In the MCT(+) group, a detectable level of IL4 in EBC and detectible levels of eicosanoids were seen in the ECT(+) group. We observed the opposite direction of ECT-induced changes in FeNO and serum tryptase concentrations in patients with detectable compared with patients without detectable levels of cytokines in EBC. We showed ECT-induced reduction in the tryptase level in patients with a nondetectable PGE2 level in EBC and an increase in tryptase levels in patients who had detectable levels of PGE2 in EBC. EBC was a useful method to estimate inflammation but only in children with symptoms and with EIB shown by a positive ECT. Children with a positive ECT had detectable levels of eicosanoids in EBC; the opposite direction of ECT-induced changes in FeNO and serum tryptase concentrations was observed. The results of above study confirm the role of mast cells and eicosanoids in the pathogenesis of EIB in children.

Biomarkers in Exhaled Breath Condensate Are Not Predictive for Pulmonary Exacerbations in Children with Cystic Fibrosis: Results of a One-Year Observational Study.

BackgroundCystic Fibrosis (CF) is characterized by chronically inflamed airways, and inflammation even increases during pulmonary exacerbations. These adverse events have an important influence on the well-being, quality of life, and lung function of patients with CF. Prediction of exacerbations by inflammatory markers in exhaled breath condensate (EBC) combined with early treatment may prevent these pulmonary exacerbations and may improve the prognosis.AimTo investigate the diagnostic accuracy of a set of inflammatory markers in EBC to predict pulmonary exacerbations in children with CF.MethodsIn this one-year prospective observational study, 49 children with CF were included. During study visits with an interval of 2 months, a symptom questionnaire was completed, EBC was collected, and lung function measurements were performed. The acidity of EBC was measured directly after collection. Inflammatory markers interleukin (IL)-6, IL-8, tumor necrosis factor α (TNF-α), and macrophage migration inhibitory factor (MIF) were measured using high sensitivity bead based flow immunoassays. Pulmonary exacerbations were recorded during the study and were defined in two ways. The predictive power of inflammatory markers and the other covariates was assessed using conditionally specified models and a receiver operating characteristic curve (SAS version 9.2). In addition, k-nearest neighbors (KNN) algorithm was applied (SAS version 9.2).ResultsSixty-five percent of the children had one or more exacerbations during the study. The conditionally specified models showed an overall correct prediction rate of 55%. The area under the curve (AUC) was equal to 0.62. The results obtained with the KNN algorithm were very similar.ConclusionAlthough there is some evidence indicating that the predictors outperform random guessing, the general diagnostic accuracy of EBC acidity and the EBC inflammatory markers IL-6, IL-8, TNF-α and MIF is low. At present it is not possible to predict pulmonary exacerbations in children with CF with the chosen biomarkers and the method of EBC analysis. The biochemical measurements of EBC markers should be improved and other techniques should be considered.

Relationships between adult asthma and oxidative stress markers and pH in exhaled breath condensate: a systematic review.

Oxidative stress has a recognized role in the pathophysiology of asthma. Recently, interest has increased in the assessment of pH and airway oxidative stress markers. Collection of exhaled breath condensate (EBC) and quantification of biomarkers in breath samples can potentially indicate lung disease activity and help in the study of airway inflammation, and asthma severity. Levels of oxidative stress markers in the EBC have been systematically evaluated in children with asthma; however, there is no such systematic review conducted for adult asthma. A systematic review of oxidative stress markers measured in EBC of adult asthma was conducted, and studies were identified by searching MEDLINE and SCOPUS databases. Sixteen papers met the inclusion criteria. Concentrations of exhaled hydrogen ions, nitric oxide products, hydrogen peroxide and 8-isoprostanes were generally elevated and related to lower lung function tests in adults with asthma compared to healthy subjects. Assessment of EBC markers may be a noninvasive approach to evaluate airway inflammation, exacerbations, and disease severity of asthma, and to monitor the effectiveness of anti-inflammatory treatment regimens. Longitudinal studies, using standardized analytical techniques for EBC collection, are required to establish reference values for the interpretation of EBC markers in the context of asthma.

Association between exhaled inflammatory markers and asthma control in children

The relationship between exhaled inflammatory markers and asthma control in children is unclear. (1) To explore the association between inflammatory markers in exhaled breath (fractional nitric oxide (FeNO), volatile organic compounds (VOCs), cytokines/chemokines) and asthma control. (2) To assess whether exhaled inflammatory markers are able to discriminate between children with persistently controlled/uncontrolled asthma. 96 asthmatic children were followed-up in a one-year observational study. Every 2 months, the following parameters were assessed: asthma control, FeNO, lung function (forced expiratory volume in 1 s (FEV1) and forced vital capacity (FVC), exhaled VOCs, and cytokines/chemokines in exhaled breath condensate (EBC). Random Forest was used to analyse the relationship between exhaled inflammatory markers and asthma control. For each model, patients were randomly selected for a training set and validation set. To assess the accuracy of the classification models, receiver operating characteristic-curves (ROC-curves) were generated. No significant association was found between the exhaled inflammatory markers (FeNO, markers in EBC, VOCs) and asthma control (area under the ROC-curve 49%). However, 15 exhaled VOCs could discriminate between subgroups of children with persistently controlled and uncontrolled asthma during all clinical visits (area under the ROC-curve 86%). Adding FeNO and markers in EBC to this model, did not lead to a more accurate classification (area under the ROC-curve 87%). There was no association between exhaled inflammatory markers and asthma control in children. However, children with persistently controlled or uncontrolled asthma during the 12 month study period could be discriminated by a set of VOCs.

Prediction of asthma exacerbations in children by innovative exhaled inflammatory markers: results of a longitudinal study.

BackgroundIn asthma management guidelines the primary goal of treatment is asthma control. To date, asthma control, guided by symptoms and lung function, is not optimal in many children and adults. Direct monitoring of airway inflammation in exhaled breath may improve asthma control and reduce the number of exacerbations.Aim1) To study the use of fractional exhaled nitric oxide (FeNO) and inflammatory markers in exhaled breath condensate (EBC), in the prediction of asthma exacerbations in a pediatric population. 2) To study the predictive power of these exhaled inflammatory markers combined with clinical parameters.Methods96 asthmatic children were included in this one-year prospective observational study, with clinical visits every 2 months. Between visits, daily symptom scores and lung function were recorded using a home monitor. During clinical visits, asthma control and FeNO were assessed. Furthermore, lung function measurements were performed and EBC was collected. Statistical analysis was performed using a test dataset and validation dataset for 1) conditionally specified models, receiver operating characteristic-curves (ROC-curves); 2) k-nearest neighbors algorithm.ResultsThree conditionally specified predictive models were constructed. Model 1 included inflammatory markers in EBC alone, model 2 included FeNO plus clinical characteristics and the ACQ score, and model 3 included all the predictors used in model 1 and 2. The area under the ROC-curves was estimated as 47%, 54% and 59% for models 1, 2 and 3 respectively. The k-nearest neighbors predictive algorithm, using the information of all the variables in model 3, produced correct predictions for 52% of the exacerbations in the validation dataset.ConclusionThe predictive power of FeNO and inflammatory markers in EBC for prediction of an asthma exacerbation was low, even when combined with clinical characteristics and symptoms. Qualitative improvement of the chemical analysis of EBC may lead to a better non-invasive prediction of asthma exacerbations.

Pulmonary inflammation in foundry workers.

To assess whether cumulative dust exposure in foundry work is associated with airway inflammation measured by the analysis of fractionated exhaled nitric oxide (NO) concentration, or by inflammatory markers in exhaled breath condensate or serum. We examined 476 dust-exposed and nonexposed foundry workers, and assessed the individual cumulative exposure to dusts and respirable quartz. Bronchial and alveolar NO production and inflammatory markers in exhaled breath condensate and in serum samples were also analyzed. After adjusting for pack-years of smoking, increased levels of alveolar NO, serum C-reactive protein, and interleukin-8 were associated with a higher level of cumulative exposure to dust. The referents had higher serum myeloperoxidase levels, bronchial NO output, and 8-isoprostane levels in exhaled breath condensate than in the dust-exposed groups. Dust exposure in foundry work may induce both systemic and alveolar inflammation.

Analysis of Short and Long Term Variability of Nasal Mucus versus Breath Condensate Inflammation Markers in Healthy Individuals

Objective: Inflammation markers were measured in nasal mucus and exhaled breath condensate (EBC) to assess the normal variability of these markers among healthy individuals. The measurement of protein markers in those matrices is valuable to provide information on inflammation endotypes and local reaction of the immune system. Methods: Inflammation markers (IFN-γ, IL-1β, IL-8, IL-10, IL-13 and TNF-α) in nasal mucus and EBC from six healthy adults were measured by Meso Scale Discovery technology. Nasal mucus was collected with sponges of polyurethane foam, which is a recently developed sampling technique. EBC was sampled via an optimized breath manoeuver with the RTUBETM device. Both collections were done within a limited sampling time of two minutes. The samples were taken on the same day and within a time period of twenty days for the determination of short and long term variability, respectively. Results: All inflammation markers were detectable in nasal mucus. This was in contrast with the measurements in EBC, in which only IFN-γ, IL-8 and IL-13 were detectable in the majority of the samples. EBC collected with the applied breathing technique gave a repeatable almost constant volume (448 μL (95% CI: 429 to 467 μL)). The collected volume of nasal mucus was more variable (from 0 to 200 μL). The variability of nasal analytes was similar over the short and long time period. The nasal inflammation marker levels were more stable within an individual than between different individuals (0.72 ≤ ICC ≤ 0.96). This was in contrast with the measurements in EBC that were highly variable over time (0.08 ≤ ICC ≤ 0.51). Conclusion: We indicate – given the good detectability and low variability of the inflammation markers determined in nasal mucus – that nasal mucus, more than EBC, is potentially a good matrix to assess inflammation in the respiratory system.

Analysis of inflammatory cytokines in human blood, breath condensate, and urine using a multiplex immunoassay platform

A change in the expression of cytokines in human biological media indicates an inflammatory response to external stressors and reflects an early step along the adverse outcome pathway (AOP) for various health endpoints. To characterize and interpret this inflammatory response, methodology was developed for measuring a suite of 10 different cytokines in human blood, exhaled breath condensate (EBC), and urine using an electrochemiluminescent multiplex Th1/Th2 cytokine immunoassay platform. Measurement distributions and correlations for eight interleukins (IL) (1β, 2, 4, 5, 8, 10, 12p70 and 13), interferon-γ (IFN-γ), and tumor necrosis factor-α (TNF-α) were evaluated using 90 blood plasma, 77 EBC, and 400 urine samples collected from nominally healthy adults subjects in North Carolina in 2008–2012. The in vivo results show that there is sufficient sensitivity for characterizing all 10 cytokines at levels of 0.05–0.10 ρg/ml with a dynamic range up to 100 ng/ml across all three of these biological media. The measured in vivo results also show that the duplicate analysis of blood, EBC and urine samples have average estimated fold ranges of 2.21, 3.49, and 2.50, respectively, which are similar to the mean estimated fold range (2.88) for the lowest concentration (0.610 ρg/ml) from a series of spiked control samples; the cytokine method can be used for all three biological media. Nine out of the 10 cytokines measured in EBC were highly correlated within one another with Spearman ρ coefficients ranging from 0.679 to 0.852, while the cytokines measured in blood had a mix of negative and positive correlations, ranging from −0.620 to 0.836. Almost all correlations between EBC and blood were positive. This work also represents the first successful within- and between-person evaluation of ultra trace-level inflammatory markers in blood, EBC, and urine.

Exhaled breath condensate as a matrix for combustion-based nanoparticle exposure and health effect evaluation.

Health assessment and medical surveillance of workers exposed to combustion nanoparticles are challenging. The aim was to evaluate the feasibility of using exhaled breath condensate (EBC) from healthy volunteers for (1) assessing the lung deposited dose of combustion nanoparticles and (2) determining the resulting oxidative stress by measuring hydrogen peroxide (H(2)O(2)) and malondialdehyde (MDA). Fifteen healthy nonsmoker volunteers were exposed to three different levels of sidestream cigarette smoke under controlled conditions. EBC was repeatedly collected before, during, and 1 and 2 hr after exposure. Exposure variables were measured by direct reading instruments and by active sampling. The different EBC samples were analyzed for particle number concentration (light-scattering-based method) and for selected compounds considered oxidative stress markers. Subjects were exposed to an average airborne concentration up to 4.3×10(5) particles/cm(3) (average geometric size ∼60-80 nm). Up to 10×10(8) particles/mL could be measured in the collected EBC with a broad size distribution (50(th) percentile ∼160 nm), but these biological concentrations were not related to the exposure level of cigarette smoke particles. Although H(2)O(2) and MDA concentrations in EBC increased during exposure, only H2O2 showed a transient normalization 1 hr after exposure and increased afterward. In contrast, MDA levels stayed elevated during the 2 hr post exposure. The use of diffusion light scattering for particle counting proved to be sufficiently sensitive to detect objects in EBC, but lacked the specificity for carbonaceous tobacco smoke particles. Our results suggest two phases of oxidation markers in EBC: first, the initial deposition of particles and gases in the lung lining liquid, and later the start of oxidative stress with associated cell membrane damage. Future studies should extend the follow-up time and should remove gases or particles from the air to allow differentiation between the different sources of H(2)O(2) and MDA.

A point-of-care testing system with Love-wave sensor and immunogold staining enhancement for early detection of lung cancer.

It has been reported that detection of exhaled breath condensate (EBC) is available for studies of pulmonary diseases, especially lung disease. In order to detect lung cancer (LC) at early stage, a point-of-care testing system suitable for measurement of tumor markers in EBC is developed. The assay, based on gold nanoparticle sandwich immunoassay and subsequent gold staining, was performed on a Love-wave sensor packaged inside a chip cartridge. Benefit from high sensitivity of Love-wave sensor, oriented immobilization of coating antibodies and immunogold staining enhancement, the present immunosensor could provide a sensitive, specific and rapid measurement. Carcinoembryonic antigen (CEA), neuron specific enolase (NSE) and squamous cell carcinoma antigen (SCC) in EBC collected from 17 patients with LC and 13 healthy volunteers were detected by this system. Results were compared with commercial chemiluminescence immunoassay and showed high correlation between two methods. Additionally, it revealed significantly statistical differences existing between two groups of subjects. These results indicate that the present system is suitable for detection of tumor markers in EBC and could be used as assistant tools for early detection of LC.

Air pollution and subclinical airway inflammation in the SALIA cohort study

BackgroundThe association between long-term exposure to air pollution and local inflammation in the lung has rarely been investigated in the general population of elderly subjects before. We investigated this association in a population-based cohort of elderly women from Germany.MethodsIn a follow-up examination of the SALIA cohort study in 2008/2009, 402 women aged 68 to 79 years from the Ruhr Area and Borken (Germany) were clinically examined. Inflammatory markers were determined in exhaled breath condensate (EBC) and in induced sputum (IS). We used traffic indicators and measured air pollutants at single monitoring stations in the study area to assess individual traffic exposure and long-term air pollution background exposure. Additionally long-term residential exposure to air pollution was estimated using land-use regression (LUR) models. We applied multiple logistic and linear regression analyses adjusted for age, indoor mould, smoking, passive smoking and socio-economic status and additionally conducted sensitivity analyses.ResultsInflammatory markers showed a high variability between the individuals and were higher with higher exposure to air pollution. NO derivatives, leukotriene (LT) B4 and tumour necrosis factor-α (TNF-α) showed the strongest associations. An increase of 9.42 μg/m3 (interquartile range) in LUR modelled NO2 was associated with measureable LTB4 level (level with values above the detection limit) in EBC (odds ratio: 1.38, 95% CI: 1.02 -1.86) as well as with LTB4 in IS (%-change: 19%, 95% CI: 7% - 32%). The results remained consistent after exclusion of subpopulations with risk factors for inflammation (smoking, respiratory diseases, mould infestation) and after extension of models with additional adjustment for season of examination, mass of IS and urban/rural living as sensitivity analyses.ConclusionsIn this analysis of the SALIA study we found that long-term exposure to air pollutants from traffic and industrial sources was associated with an increase of several inflammatory markers in EBC and in IS. We conclude that long-term exposure to air pollution might lead to changes in the inflammatory marker profile in the lower airways in an elderly female population.