Occupational asthma and allergic diseases of the upper airway play an important role in occupational diseases. The most frequent allergens, which causes occupational respiratory diseases in the Czech Republic, are isocyanates and wheat and rye flour. Isocyanates are the substances used in car industry during production of polyurethanes, they are in colours and glues. Occupational allergic asthma and rhinitis are diagnosed using specific challenge tests with substances from the workplace. These tests can be done in a special exposure box in a lab with substances taken by hygiene specialists from the patient´s workplace or using direct exposure at the workplace. Spirometry and anterior active rhinomanometry are used as objective methods during these tests. For prognosis the early diagnosis and withdrawal from the occupational allergen exposure are essential. Keywords: allergic asthma, allergic rhinitis, irritant-induced asthma, irritant-induced rhinitis, isocyanate, occupational disease, specific inhalation challenge.

• Klíčová slova
• allergic asthma, allergic rhinitis, irritant-induced asthma, irritant-induced rhinitis, isocyanate, occupational disease, specific inhalation challenge,
• MeSH
• lidé MeSH
• pracovní expozice * MeSH
• profesionální astma * diagnóza   terapie   MeSH
• rýma * diagnóza   etiologie   terapie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

OBJECTIVES: To search for optimal markers in the exhaled breath condensate (EBC), plasma and urine that would reflect the activity/severity of occupational asthma (OA) after the withdrawal from the exposure to the allergen. MATERIAL AND METHODS: Markers of oxidative stress: 8-iso-prostaglandin F2α (8-isoprostane, 8-ISO), malondialdehyde (MDA), 4-hydroxy-trans-2-nonenale (HNE), cysteinyl leukotrienes (LT) and LTB4 were determined using liquid chromatography and mass spectrometry in 43 subjects with immunological OA (49.3 ± 11.8 years), removed from the exposure to the sensitizing agent 10.5 ± 6.5 years ago; and in 20 healthy subjects (49.0 ± 14.9 years). EBC was harvested both before and after the methacholine challenge test. In parallel, identical markers were collected in plasma and urine. The results were analyzed together with forced expiratory volume in one second (FEV1), blood eosinophils, immunoglobulin E (IgE) and eosinophilic cationic protein (ECP) and statistically evaluated (Spearman rank correlation rS, two- or one-sample t tests and alternatively Kruskal Wallis or pair Wilcoxon tests). RESULTS: Several parameters of lung functions were lower in the patients (FEV1% predicted, MEF25% and MEF50%, Rtot%, p < 0.001). Shorter time interval since the removal from the allergen exposure correlated with higher ECP (rS = 0.375) and lower FEV1%, MEF25% and MEF50% after methacholine challenge (rS = -0.404, -0.425 and -0.532, respectively). In the patients, IgE (p < 0.001) and ECP (p = 0.009) was increased compared to controls. In EBC, 8-ISO and cysteinyl LTs were elevated in the asthmatics initially and after the challenge. Initial 8-ISO in plasma correlated negatively with FEV1 (rS = -0.409) and with methacholine PD20 (rS = -0.474). 8-ISO in plasma after the challenge correlated with IgE (rS = 0.396). CONCLUSIONS: The improvement in OA is very slow and objective impairments persist years after removal from the exposure. Cysteinyl LTs and 8-ISO in EBC and 8-ISO in plasma might enrich the spectrum of useful objective tests for the follow-up of OA.

• MeSH
• biologické markery analýza   metabolismus   MeSH
• cystein analýza   MeSH
• dechové testy MeSH
• dinoprost analogy a deriváty   analýza   krev   MeSH
• dospělí MeSH
• eozinofilní kationtový protein krev   MeSH
• leukotrieny analýza   MeSH
• lidé středního věku MeSH
• lidé MeSH
• maximální exspirační průtok ve střední části MeSH
• následné studie MeSH
• profesionální astma metabolismus   patofyziologie   MeSH
• studie případů a kontrol MeSH
• stupeň závažnosti nemoci MeSH
• usilovný výdechový objem MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 8-epi-prostaglandin F2alpha MeSH Prohlížeč
• biologické markery MeSH
• cystein MeSH
• cysteinyl-leukotriene MeSH Prohlížeč
• dinoprost MeSH
• eozinofilní kationtový protein MeSH
• leukotrieny 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

Exhaled breath condensate (EBC) is assumed to reflect processes in the lungs, yet it is unknown whether oxidative stress markers in EBC are affected by systemic disorders (atherosclerosis, hypertension, diabetes) or whether lung diseases increase markers in plasma and urine. 8-isoprostane, 4-hydroxy-trans-2-nonenale (HNE) and malondialdehyde (MDA) were measured using liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS/MS) in EBC, plasma and urine in 82 patients (45 with asbestosis and hyalinosis, and 37 with silicosis) and in 29 control subjects. 8-isoprostane and HNE in EBC, and HNE in urine were higher in both groups of patients. In addition, 8-isoprostane in plasma and urine, and MDA in urine were higher in asbestos-exposed patients and MDA in plasma in silicotics, with this marker in plasma correlated with the grade of silicosis. In all subjects, 8-isoprostane in EBC correlated with urine (r=0.38, p<0.001) and plasma levels (r=0.28, p=0.003), and HNE and MDA with urine levels (r=0.31, p<0.001; r=0.23, p=0.016, respectively). Most markers positively correlated with lung function impairment, EBC markers negatively with vitamin E supplementation. To conclude: The influence of satisfactorily controlled systemic disorders on markers in EBC in patients with pneumoconioses is not significant. In addition to oxidative stress markers in EBC, lung fibroses may increase oxidative stress markers in plasma and urine.

• MeSH
• biologické markery krev   moč   MeSH
• dechové testy MeSH
• fibróza diagnóza   patofyziologie   MeSH
• lidé MeSH
• oxidační stres fyziologie   MeSH
• plicní nemoci diagnóza   MeSH
• senioři MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• biologické markery MeSH

A rapid and precise method for the identification and quantification of cysteinyl leukotrienes (leukotriene C(4), leukotriene D(4) and leukotriene E(4)), essential markers of bronchial asthma, in exhaled breath condensate was developed. The protocol consists of immunoaffinity separation and a detection step, liquid chromatography combined with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). In particular, the selected reaction monitoring mode was used for its extremely high degree of selectivity and the stable-isotope-dilution assay for its high precision of quantification. The developed method was characterized with a high precision (≤ 7.7%, determined as RSD), an acceptable accuracy (90.4-93.7%, determined as recovery), a low limit of detection (≤ 2 pg/ml EBC) and a low limit of quantification (≤ 10 pg/ml EBC). It was compared to other simple, clinically appropriate combinations of pre-treatment methods (solid phase extraction and lyophilization) with LC/MS. Finally, the method (a combination of immunoaffinity separation with LC-MS) was successfully tested in a clinical study where a significant difference was found in the concentration levels of cysteinyl leukotrienes between patients with occupational bronchial asthma and healthy subjects.

• MeSH
• bronchiální astma diagnóza   MeSH
• chromatografie kapalinová metody   MeSH
• cystein analýza   MeSH
• dechové testy metody   MeSH
• hmotnostní spektrometrie s elektrosprejovou ionizací metody   MeSH
• imunoanalýza metody   MeSH
• leukotrieny analýza   MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• vydechnutí MeSH
• Check Tag
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cystein MeSH
• cysteinyl-leukotriene MeSH Prohlížeč
• leukotrieny MeSH

Sensitive assay method was developed for a parallel, rapid and precise determination of the most prominent oxidative stress biomarkers: 8-iso-prostaglandin F(2alpha), malondialdehyde and 4-hydroxynonenal. The method consisted of a pre-treatment part a solid-phase extraction, for rapid and effective isolation of biomarkers from body fluids (exhaled breath condensate, plasma and urine) and the detection method LC-ESI-MS/MS, where the selected reaction monitoring mode was used for its extremely high degree of selectivity and the stable-isotope-dilution assay for its high precision of quantification. The developed method was characterized by the following parameters: the imprecision was below 14.3%, the mean inaccuracy was determined to be lower than 13.1%. The method was tested on samples obtained from patients diagnosed with asbestosis, pleural hyalinosis or silicosis, i.e. occupational lung diseases caused by fibrogenic dusts, inducing oxidative stress in the respiratory system, and then compared to samples from healthy subjects. The difference in concentration levels of biomarkers between the two groups was perceptible in all the body fluids (the difference observed in an exhaled breath condensate was statistically most significant).

• MeSH
• aldehydy analýza   krev   moč   MeSH
• azbest toxicita   MeSH
• biologické markery analýza   krev   moč   MeSH
• dinoprost analogy a deriváty   analýza   krev   moč   MeSH
• extrakce na pevné fázi metody   MeSH
• lidé středního věku MeSH
• lidé MeSH
• malondialdehyd analýza   krev   moč   MeSH
• oxid křemičitý toxicita   MeSH
• oxidační stres * MeSH
• plicní nemoci krev   etiologie   metabolismus   moč   MeSH
• senioři MeSH
• studie případů a kontrol MeSH
• tandemová hmotnostní spektrometrie metody   MeSH
• tělesné tekutiny chemie   metabolismus   MeSH
• vysokoúčinná kapalinová chromatografie metody   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• senioři MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 4-hydroxy-2-nonenal MeSH Prohlížeč
• 8-epi-prostaglandin F2alpha MeSH Prohlížeč
• aldehydy MeSH
• azbest MeSH
• biologické markery MeSH
• dinoprost MeSH
• malondialdehyd MeSH
• oxid křemičitý MeSH

Between 1992 and 2005, 72 cases of occupational hypersensitive pneumonitis were reported to the Czech National Registry of Occupational Diseases. This represented 0.24% of all occupational diseases reported in the Czech Republic during that period. The greatest number of cases occurred in 1997 (10 cases), of which men constituted 58.3% (42 cases) and women 41.7% (30 cases). The most prevalent cases were farmer's lung (50 cases), malt worker's lung (7 cases) and chemical worker's lung (6 cases). Agriculture was the most common economic activity (total 48 cases), with cattleman and dairyman (total 26 cases) the most frequent occupations; less common were tractor driver (8 cases) and maltster (7 cases). Typical case reports after different exposures are presented. A peak in frequency was observed in the age groups of 45-49 and 50-54 yr (20.8% and 19.4% of cases, respectively) and within the first four years of employment (22.2% of cases). Median age was 48 yr and median exposure 12.5 yr. Incidence was in the range of 0.00-0.20 per 100,000 workers, which appears rather low. Due to the difficult diagnostics of hypersensitive pneumonitis, the actual number of cases is undoubtedly higher, and this disease is probably under-reported.

• MeSH
• dospělí MeSH
• hypersenzitivní pneumonitida epidemiologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• pracovní expozice * MeSH
• registrace * MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH

Asbestosis and pleural plaques exhibit unpredictable but progressive development, and there are no markers routinely available to measure their prognosis. Asbestos exposure induces the generation of reactive oxygen species, and 8-isoprostane is involved in experimental asbestos-related lung toxicity. This oxidative stress marker was measured in exhaled breath condensate (EBC) in 92 former asbestos workers with mean age 68.8+/-1.7 yr and mean duration of asbestos exposure 24.1+/-2.0 yr. The control group had 46 subjects with mean age 65.2+/-3.3 yr. The mean level of 8-isoprostane, analyzed by liquid chromatography-electrospray ionization-mass spectrometry, was higher in asbestos-exposed subjects (69.5+/-6.6 pg/ml, p=0.0001) compared with the control group, where the concentration was 47.0+/-7.8 pg/ml. The results presented support the hypothesis that oxidative stress due to asbestos is the main cause of increased 8-isoprostane in EBC. Measurement of 8-isoprostane in EBC is a promising non-invasive means for assessing the activity of asbestos-induced diseases.

• MeSH
• azbest metabolismus   otrava   MeSH
• azbestóza metabolismus   MeSH
• biologické markery analýza   MeSH
• dechové testy metody   MeSH
• dinoprost analogy a deriváty   analýza   metabolismus   MeSH
• hodnocení rizik MeSH
• lidé středního věku MeSH
• lidé MeSH
• oxidační stres MeSH
• pracovní expozice MeSH
• prognóza MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• studie případů a kontrol MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 8-epi-prostaglandin F2alpha MeSH Prohlížeč
• azbest MeSH
• biologické markery MeSH
• dinoprost MeSH

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

Rapid and precise method for the determination of 8-iso-prostaglandin F(2alpha), an essential marker of the oxidative stress, in exhaled breath condensate (EBC) was developed. The protocol consisted of stable isotope dilution, immunoseparation combined with selective and sensitive LC-ESI-MS/MS operated in multiple reaction monitoring (MRM) mode. The imprecision of the developed method was below 8.8%, the parameter of mean inaccuracy was determined as <9.6% (0-250pg of 8-iso-prostaglandin F(2alpha)/ml EBC). The limit of detection (LOD) was 1 pg/ml EBC and limit of quantification (LOQ) 5 pg/ml EBC. A significant difference in 8-iso-prostaglandin F(2alpha) content between the group of asbestosis patients and healthy volunteers was found.

• MeSH
• dechové testy * MeSH
• dinoprost analogy a deriváty   analýza   MeSH
• hmotnostní spektrometrie s elektrosprejovou ionizací metody   MeSH
• kalibrace MeSH
• lidé MeSH
• reprodukovatelnost výsledků MeSH
• senzitivita a specificita MeSH
• tandemová hmotnostní spektrometrie metody   MeSH
• vysokoúčinná kapalinová chromatografie metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• validační studie MeSH
• Názvy látek
• 8-epi-prostaglandin F2alpha MeSH Prohlížeč
• dinoprost MeSH