Researchers in nanocomposite processing may inhale a variety of chemical agents, including nanoparticles. This study investigated airway oxidative stress status in the exhaled breath condensate (EBC). Nineteen employees (42.4 ± 11.4 y/o), working in nanocomposites research for 18.0 ± 10.3 years were examined pre-shift and post-shift on a random workday, together with nineteen controls (45.5 ± 11.7 y/o). Panels of oxidative stress biomarkers derived from lipids, nucleic acids, and proteins were analyzed in the EBC. Aerosol exposures were monitored during three major nanoparticle generation operations: smelting and welding (workshop 1) and nanocomposite machining (workshop 2) using a suite of real-time and integrated instruments. Mass concentrations during these operations were 0.120, 1.840, and 0.804 mg/m³, respectively. Median particle number concentrations were 4.8 × 10⁴, 1.3 × 10⁵, and 5.4 × 10⁵ particles/cm³, respectively. Nanoparticles accounted for 95, 40, and 61%, respectively, with prevailing Fe and Mn. All markers of nucleic acid and protein oxidation, malondialdehyde, and aldehydes C₆⁻C13 were elevated, already in the pre-shift samples relative to controls in both workshops. Significant post-shift elevations were documented in lipid oxidation markers. Significant associations were found between working in nanocomposite synthesis and EBC biomarkers. More research is needed to understand the contribution of nanoparticles from nanocomposite processing in inducing oxidative stress, relative to other co-exposures generated during welding, smelting, and secondary oxidation processes, in these workshops.

• Klíčová slova
• exhaled breath condensate, inhalation, nanocomposites, nanoparticles, occupational exposure, oxidative stress, workers,
• Publikační typ
• časopisecké články MeSH

Analysis of Exhaled breath condensate (EBC) is a re-discovered approach to monitoring the course of the disease and reduce invasive methods of patient investigation. However, the major disadvantage and shortcoming of the EBC is lack of reliable and reproducible standardization of the method. Despite many articles published on EBC, until now there is no clear consensus on whether the analysis of EBC can provide a clue to diagnosis of the diseases. The purpose of this paper is to investigate our own method, to search for possible standardization and to obtain our own initial experience. Thirty healthy volunteers provided the EBC, in which we monitored the density, pH, protein, chloride and urea concentration. Our results show that EBC pH is influenced by smoking, and urea concentrations are affected by the gender of subjects. Age of subjects does not play a role. The smallest coefficient of variation between individual volunteers is for density determination. Current limitations of EBC measurements are the low concentration of many biomarkers. Standardization needs to be specific for each individual biomarker, with focusing on optimal condensate collection. EBC analysis has a potential become diagnostic test, not only for lung diseases.

• Klíčová slova
• exhaled breath condensate, healthy subjects, standardization,
• MeSH
• biologické markery analýza   metabolismus   MeSH
• chloridy analýza   metabolismus   MeSH
• dechové testy metody   MeSH
• dospělí MeSH
• koncentrace vodíkových iontů MeSH
• kouření metabolismus   MeSH
• lidé středního věku MeSH
• lidé MeSH
• močovina analýza   metabolismus   MeSH
• odběr biologického vzorku MeSH
• pilotní projekty MeSH
• proteiny analýza   metabolismus   MeSH
• referenční hodnoty MeSH
• referenční standardy MeSH
• senioři MeSH
• sexuální faktory MeSH
• věkové faktory MeSH
• zdraví dobrovolníci pro lékařské studie MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• biologické markery MeSH
• chloridy MeSH
• močovina MeSH
• proteiny MeSH

Exhaled breath condensate (EBC) is an attractive, non-invasive sample for clinical diagnostics. During EBC collection, its composition is influenced by the collection temperature, a factor that is often not thoroughly monitored and controlled. In this study, we assembled a novel, simple, portable, and inexpensive device for EBC collection, able to maintain a stable temperature at any value between -7 °C and +12 °C. The temperature was controlled using a microcontroller and a thermoelectric cooler that was employed to cool the aluminum block holding the glass tube or the polypropylene syringe. The performance of the novel sampler was compared with the passively cooled RTube™ and a simple EBC sampler, in which the temperature was steadily increasing during sampling. The developed sampler was able to maintain a stable temperature within ±1 °C. To investigate the influence of different sampling temperatures (i.e., +12, -7, -80 °C) on the analyte content in EBC, inorganic ions and organic acids were analyzed by capillary electrophoresis with a capacitively coupled contactless conductivity detector. It was shown that the concentration of metabolites decreased significantly with decreasing temperature. The portability and the ability to keep a stable temperature during EBC sampling makes the developed sampler suitable for point-of-care diagnostics.

• Klíčová slova
• Arduino, Capillary electrophoresis, Collection device, Exhaled breath condensate, Ion analysis, Temperature control,
• MeSH
• biologické markery MeSH
• dechové testy * MeSH
• elektroforéza kapilární MeSH
• point of care testing MeSH
• teplota MeSH
• vydechnutí * MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• biologické markery MeSH

Exhaled breath condensate is a promising, non-invasive, diagnostic sample obtained by condensation of exhaled breath. Starting from a historical perspective of early attempts of breath testing towards the contemporary state-of-the-art breath analysis, this review article focuses mainly on the progress in determination of non-volatile compounds in exhaled breath condensate. The mechanisms by which the aerosols/droplets of non-volatile compounds are formed in the airways are discussed with methodological consequences for sampling. Dilution of respiratory droplets is a major problem for correct clinical interpretation of the measured data and there is an urgent need for standardization of EBC. This applies also for collection instrumentation and therefore various commercial and in-house built devices are described and compared with regard to their design, function and collection parameters. The analytical techniques and methods for determination of non-volatile compounds as potential markers of oxidative stress and lung inflammation are scrutinized with an emphasis on method suitability, sensitivity and appropriateness. The relevance of clinical findings for each group of possible non-volatile markers of selected pulmonary diseases and methodological recommendations with emphasis on interdisciplinary collaboration that is essential for future development into a fully validated clinical diagnostic tool are given.

• Klíčová slova
• 3-NT, 3-nitrotyrosine, 5-HPETE, 5-LO, 5-hydroperoxyeicosatetraenoic acid, 5-lipoxygenase, 8-IP, 8-isoprostane, ALF, Analytical techniques, BAL, CF, COPD, COX, EBC, Exhaled breath condensate (EBC), FDA, GSH, GSSG, INF-γ, Ions, LT, Metabolites, NMR, PCA, PE, PG, PLS, PLS-DA, PP, PTFE, Proteins, ROS, RS–NO, S-nitrosothiols, SI, TNF-α, U.S. Food and Drug Administration, VOC, airway lining fluid, bronchoalveolar lavage, chronic obstructive pulmonary disease, cyclooxygenase, cystic fibrosis, exhaled breath condensate, glutathione, glutathione disulfide, interferon γ, leukotriene, nuclear magentic resonance, partial least squares, partial least squares discriminant analysis, polyethylene, polypropylene, polytetrafluoroethylene, principal component analysis, prostaglandin, reactive oxygen species, sputum induction, tumor necrosis factor α, volatile organic compounds,
• MeSH
• biologické markery analýza   metabolismus   MeSH
• dechové testy přístrojové vybavení   metody   MeSH
• imunoanalýza MeSH
• lidé MeSH
• plicní nemoci diagnóza   metabolismus   MeSH
• plynová chromatografie s hmotnostně spektrometrickou detekcí MeSH
• reaktivní formy dusíku analýza   metabolismus   MeSH
• vzduch analýza   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• biologické markery MeSH
• reaktivní formy dusíku MeSH

BACKGROUND: Inflammatory markers in exhaled breath condensate (EBC) are investigated as a non-invasive approach to monitoring of inflammation in the respiratory tract. EBC concentrations of nitrite and nitrate, the stable end products of oxidative metabolism of nitric oxide, are increased in patients with asthma, especially during acute exacerbations. OBJECTIVES: To examine methodological aspects of nitrite and nitrate measurements in EBC such as sample collection, storage and analysis. METHODS: In a randomized study, EBC was collected twice within 1 h (with and without a nose clip) in 20 healthy adults and 20 patients with well-controlled asthma and no symptoms of allergic rhinitis. Nitrite and nitrate were assayed by ionex chromatography and fluorimetrically after derivatization with diaminonaphthalene. RESULTS: The geometric mean [exp (mean +/- SD)] EBC levels of nitrite and nitrate in healthy subjects [4.3 (3.0-6.1) and 11.0 (5.3-22.7) micromol/l] and patients [4.6 (2.6-7.3) and 8.7 (3.2-23.8) micromol/l] did not differ (p = 0.13). Wearing a nose clip (p = 0.3) did not influence nitrite and nitrate concentrations. The mean intra-subject %CVs of EBC concentrations of nitrite were 26 and 21% in healthy subjects and patients, while those of nitrate achieved 49 and 88%, respectively. CONCLUSIONS: Ionex chromatography of nitrite and nitrate requires no sample pretreatment and provides comparable results as a more laborious diaminonaphthalene method. EBC samples should be kept cold (8 degrees C) and analyzed for nitrite and nitrate within 24 h of collection or stored in the freezer and thawed preferably only once. Wearing a nose clip during EBC collection has no influence on nitrite and nitrate concentrations. Short-term repeatability of nitrite and nitrate measurements was worse compared to published data on exhaled nitric oxide.

• MeSH
• bronchiální astma metabolismus   MeSH
• chromatografie MeSH
• dechové testy přístrojové vybavení   metody   MeSH
• dospělí MeSH
• dusičnany metabolismus   MeSH
• dusitany metabolismus   MeSH
• fluorometrie MeSH
• klinické křížové studie MeSH
• lidé MeSH
• odběr biologického vzorku MeSH
• plíce metabolismus   MeSH
• vydechnutí * MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• randomizované kontrolované studie MeSH
• validační studie MeSH
• Názvy látek
• dusičnany MeSH
• dusitany MeSH

Leukotrienes (LTs) are involved in the pathogenesis of lung fibrosis and were increased in exhaled breath condensate (EBC) of the patients with pneumoconiosis. However the possible influence of extra-pulmonary disorders on the EBC markers is not known. Therefore in parallel with EBC, LTs' levels in the plasma and urine were measured in patients with pneumoconiosis (45 × asbestos exposure, 37 × silica exposure) and in 27 controls. Individual LTs B4, C4, D4 and E4 were measured by liquid chromatography - electrospray ionization - tandem mass spectrometry (LC-ESI-MS/MS). In EBC, LT D4 and LT E4 were increased in both groups of patients (p<0.001 and p<0.05), comparing with the controls. Both LT B4 and cysteinyl LTs were elevated in asbestos-exposed subjects (p<0.05). Asbestosis with more severe radiological signs (s1/s2-t3/u2) and lung functions impairment has shown higher cysteinyl LTs and LT C4 in the EBC (p<0.05) than mild asbestosis (s1/s0-s1/s1). In addition, in the subjects with asbestosis, cysteinyl LTs in EBC correlated with TLC (-0.313, p<0.05) and TLCO/Hb (-0.307, p<0.05), and LT C4 with TLC (-0.358, p<0.05). In pneumoconioses, EBC appears the most useful from the 3 fluids studied.

• MeSH
• azbestóza diagnostické zobrazování   metabolismus   MeSH
• dechové testy * MeSH
• leukotrien B4 analýza   krev   moč   MeSH
• leukotrien C4 analýza   krev   moč   MeSH
• leukotrien D4 analýza   krev   moč   MeSH
• leukotrien E4 analýza   krev   moč   MeSH
• leukotrieny analýza   krev   moč   MeSH
• lidé středního věku MeSH
• lidé MeSH
• radiografie MeSH
• respirační funkční testy MeSH
• senioři MeSH
• silikóza diagnostické zobrazování   metabolismus   MeSH
• stupeň závažnosti nemoci MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• leukotrien B4 MeSH
• leukotrien C4 MeSH
• leukotrien D4 MeSH
• leukotrien E4 MeSH
• leukotrieny MeSH

Nanoscale titanium dioxide (nanoTiO2) is a commercially important nanomaterial. Animal studies have documented lung injury and inflammation, oxidative stress, cytotoxicity and genotoxicity. Yet, human health data are scarce and quantitative risk assessments and biomonitoring of exposure are lacking. NanoTiO2 is classified by IARC as a group 2B, possible human carcinogen. In our earlier studies we documented an increase in markers of inflammation, as well as DNA and protein oxidative damage, in exhaled breath condensate (EBC) of workers exposed nanoTiO2. This study focuses on biomarkers of lipid oxidation. Several established lipid oxidative markers (malondialdehyde, 4-hydroxy-trans-hexenal, 4-hydroxy-trans-nonenal, 8-isoProstaglandin F2α and aldehydes C6-C12) were studied in EBC and urine of 34 workers and 45 comparable controls. The median particle number concentration in the production line ranged from 1.98 × 104 to 2.32 × 104 particles/cm3 with ∼80% of the particles <100 nm in diameter. Mass concentration varied between 0.40 and 0.65 mg/m3. All 11 markers of lipid oxidation were elevated in production workers relative to the controls (p < 0.001). A significant dose-dependent association was found between exposure to TiO2 and markers of lipid oxidation in the EBC. These markers were not elevated in the urine samples. Lipid oxidation in the EBC of workers exposed to (nano)TiO2 complements our earlier findings on DNA and protein damage. These results are consistent with the oxidative stress hypothesis and suggest lung injury at the molecular level. Further studies should focus on clinical markers of potential disease progression. EBC has reemerged as a sensitive technique for noninvasive monitoring of workers exposed to engineered nanoparticles.

• Klíčová slova
• Nanoparticles, TiO2, aldehydes, exhaled breath condensate, monitoring, occupational exposure, oxidative stress,
• MeSH
• biologické markery analýza   moč   MeSH
• chemický průmysl MeSH
• dechové testy MeSH
• dinoprost analogy a deriváty   analýza   moč   MeSH
• lidé MeSH
• malondialdehyd analýza   moč   MeSH
• metabolismus lipidů MeSH
• monitorování životního prostředí metody   MeSH
• nanočástice analýza   toxicita   MeSH
• oxidace-redukce MeSH
• oxidační stres účinky léků   MeSH
• peroxidace lipidů účinky léků   MeSH
• poškození DNA MeSH
• pracovní expozice škodlivé účinky   analýza   MeSH
• spektrofotometrie atomová MeSH
• titan analýza   toxicita   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• 8-epi-prostaglandin F2alpha MeSH Prohlížeč
• biologické markery MeSH
• dinoprost MeSH
• malondialdehyd MeSH
• titan MeSH
• titanium dioxide MeSH Prohlížeč

Human health data regarding exposure to nanoparticles are extremely scarce and biomonitoring of exposure is lacking in spite of rodent pathological experimental data. Potential markers of the health-effects of engineered nanoparticles were examined in 30 workers exposed to TiO2 aerosol, 22 office employees of the same plant, and 45 unexposed controls. Leukotrienes (LT) B4, C4, E4, and D4 were analysed in the exhaled breath condensate (EBC) and urine via liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). Fractional exhaled nitric oxide (FeNO) and spirometry was also measured. The median particle number concentration of the aerosol in the production ranged from 1.98 × 10(4) to 2.32 × 10(4) particles cm(-3); about 80% of the particles were <100 nm in diameter. Median total mass concentration varied between 0.4 and 0.65 mg m(-3). All LT levels in workers' EBC were elevated relative to the controls (p < 0.01). LTs in the EBC sample were correlated with titanium levels. Urinary LTs were not elevated in the workers and office employees. Office workers had higher LTB4 in EBC (p < 0.05), and higher levels of FeNO (p < 0.01). FeNO was higher in office employees with allergic diseases and was negatively correlated with smoking (p < 0.01). In spirometry significant impairment in the workers was seen only for %VCIN and %PEF (both p < 0.01). Multiple regression analysis confirmed a significant association between production of TiO2 and all cysteinyl LTs in EBC (p < 0.01) and impaired %VCIN and %PEF (both p < 0.01). LTB4 was also associated with smoking (p < 0.01). LT levels complemented our earlier findings of DNA, protein, and lipid damage in the EBC of workers with nanoTiO2 exposures. Cysteinyl LTs in EBC analysis suggest inflammation and potential fibrotic changes in the lungs; they may be helpful for monitoring the biological effect of (nano)TiO2 on workers. Spirometry was not sensitive enough.

• MeSH
• aerosoly analýza   MeSH
• biologické markery analýza   MeSH
• dechové testy metody   MeSH
• dospělí MeSH
• koncentrace vodíkových iontů MeSH
• leukotrieny analýza   moč   MeSH
• lidé MeSH
• nanočástice škodlivé účinky   MeSH
• oxid dusnatý analýza   MeSH
• pracoviště MeSH
• pracovní expozice analýza   MeSH
• regresní analýza MeSH
• respirační funkční testy MeSH
• studie případů a kontrol MeSH
• tandemová hmotnostní spektrometrie MeSH
• titan škodlivé účinky   MeSH
• vydechnutí * MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aerosoly MeSH
• biologické markery MeSH
• leukotrieny MeSH
• oxid dusnatý MeSH
• titan MeSH
• titanium dioxide MeSH Prohlížeč

Exhaled breath condensate (EBC) contains many substances, which could help in diagnosis of occupational asthma. The aim of the study is to monitor leukotrienes (LT) and 8-isoprostane from EBC in bronchoprovocation tests with allergens in 47 patients with suspected occupational asthma. Forty-one patients were tested negative. In negative bronchoprovocation tests, no significant differences (P<0.05) were seen between the five measurements during and after the test. In control measurements (without provocation), significant differences were found among four measurements done within 24h for 8-isoprostane (P=0.0138). The relationship between the log transformed ratios of the EBC parameters and FEV(1) was never significant at the 5% level in control measurements, while in negative tests, statistical significance was recorded for LTB(4) (P=0.0299) before and 5h after the test. Six of 47 patients were tested positive. Such a small number of patients did not allow proper statistical analysis and therefore, the results are described separately for each patient.

• MeSH
• alergeny imunologie   MeSH
• dechové testy MeSH
• dinoprost analogy a deriváty   analýza   metabolismus   MeSH
• dospělí MeSH
• látky znečišťující vzduch v pracovním prostředí imunologie   MeSH
• leukotrieny analýza   metabolismus   MeSH
• lidé MeSH
• vydechnutí fyziologie   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• 8-epi-prostaglandin F2alpha MeSH Prohlížeč
• alergeny MeSH
• dinoprost MeSH
• látky znečišťující vzduch v pracovním prostředí MeSH
• leukotrieny MeSH

Human data concerning exposure to nanoparticles are very limited, and biomarkers for monitoring exposure are urgently needed. In a follow-up of a 2016 study in a nanocomposites plant, in which only exhaled breath condensate (EBC) was examined, eight markers of oxidative stress were analyzed in three bodily fluids, i.e., EBC, plasma and urine, in both pre-shift and post-shift samples in 2017 and 2018. Aerosol exposures were monitored. Mass concentration in 2017 was 0.351 mg/m3 during machining, and 0.179 and 0.217 mg/m3 during machining and welding, respectively, in 2018. In number concentrations, nanoparticles formed 96%, 90% and 59%, respectively. In both years, pre-shift elevations of 50.0% in EBC, 37.5% in plasma and 6.25% in urine biomarkers were observed. Post-shift elevation reached 62.5% in EBC, 68.8% in plasma and 18.8% in urine samples. The same trend was observed in all biological fluids. Individual factors were responsible for the elevation of control subjects' afternoon vs. morning markers in 2018; all were significantly lower compared to those of workers. Malondialdehyde levels were always acutely shifted, and 8-hydroxy-2-deoxyguanosine levels best showed chronic exposure effect. EBC and plasma analysis appear to be the ideal fluids for bio-monitoring of oxidative stress arising from engineered nanomaterials. Potential late effects need to be targeted and prevented, as there is a similarity of EBC findings in patients with silicosis and asbestosis.

• Klíčová slova
• biomarkers, controls, exhaled breath condensate, nanoparticles, oxidative stress, plasma, urine, workers,
• Publikační typ
• časopisecké články MeSH