AAethodo ogoica improvements for measuring eicosanoids and cytokines in exhaled breath condensate

ArticleinRespiratory Medicine 100(1):34-8 · February 2006with28 Reads
DOI: 10.1016/j.rmed.2005.04.007 · Source: PubMed
Abstract
Exhaled breath condensate (EBC) is simple to collect and as such a non-invasive method that has attracted substantial interest in the last few years. However, several methodological concerns have been raised and it has been difficult to reproduce results between different centres. Because of low concentrations of inflammatory markers, potential loss in the sampling system may have great influence. The aim of the present study was to se if evaporation and plastic coating could facilitate detection. Through methodological improvements, we have now made it possible to measure EBC concentrations of eicosanoids and cytokines in our system. Due to absorbance of both fatty acid derivates and proteins to several plastics, the first step is coating of all surfaces with bovine serum albumin and Tween 20. Since several assays are sensitive to these factors, the methodology has to be standardised to avoid false results. Secondly, larger amounts of EBC have to be vacuum-dried, and thereafter resolved in the respective assay buffers. The EBCs have to be concentrated 5-10 times, depending on samples and assay sensitivity. Due to these improvements we can measure, for example, cysteinyl-leukotrienes, leukotriene B4, prostaglandin E and 8-isoprostane. High sensitivity assays have also made it possible to measure cytokines, for example, interleukin (IL)-1beta, IL-8 and IL-13. We are aware of different results from other labs. However, it seems essential to coat and to concentrate the samples in order to achieve reliable and measurable results.
    • "Exhaled breath condensate (EBC). EBC was collected using a breath condenser (EuroScreen; Jaeger, Wuerzburg, Germany) according to existing recommendations [20]. In order to avoid loss of molecules from inflammatory markers to the surface of the sampling tubes, the tubes were coated with 1% bovine serum albumin and 0.01% Tween 20 for 30 minutes. "
    [Show abstract] [Hide abstract] ABSTRACT: Introduction and objective: Swine confinement buildings are known to contain large concentrations of airway irritants, and a number of studies have shown acute inflammatory effects in previously unexposed subjects when introduced to the environment in such buildings. However, studies comparing different methods of assessing such reactions are lacking, and it is not known if a measurable response could be found at lower exposure rates. The purpose of this study was to investigate exposure levels in a Norwegian swine confinement building, the airway response to such exposure, and to compare invasive and non-invasive methods of response measurement. Materials and method: Twelve medical students who were previously unexposed to swine dust stayed in a swine confinement building in Norway for 4 hours, and underwent measurements before and after the start of exposure. The same measurements were also performed beforehand, on the same weekday without exposure, in such a manner that the subjects were their own controls. Results: The exposure assessment showed considerably lower concentrations of organic dust and endotoxin than earlier studies. However, small, but significant increases in markers of airway inflammation, were found. Conclusions: Airway response can be measured after lower exposure to airborne irritants in swine confinement buildings than previously known. Further research is needed to assess whether this finding can be utilized for preventive purposes.
    Full-text · Article · Sep 2014
    • "In contrast to the numerous small molecule biomarkers identified, such as leukotrienes (LTB 4 , CysLT) and nitric oxide (NO), proteomic analysis of EBC has had a limited number of potential protein biomarkers identified due to methodological and instrumental hurdles[4,8,32,33]. The analytical difficulties of EBC proteomics can be attributed to several factors including, low protein concentration, inconsistent sample preparation and sample loss and instrumental sensitivities343536. Although present, the limitations surrounding EBC proteomics can be potentially overcome. "
    [Show abstract] [Hide abstract] ABSTRACT: Exhaled breath condensate (EBC) has been established as a potential source of respiratory biomarkers. Compared to the numerous small molecules identified, the protein content of EBC has remained relatively unstudied due to the methodological and technical difficulties surrounding EBC analysis. In this review, we discuss the proteins identified in EBC, by mass spectrometry, focusing on the significance of those proteins identified. We will also review the limitations surrounding mass spectral EBC protein analysis emphasizing recommendations to enhance EBC protein identifications by mass spectrometry. Finally, we will provide insight into the future directions of the EBC proteomics field.
    Full-text · Article · Sep 2014
    • "Recently the first results of study researches of these processes in biological fluids such as exhaled breath condensate or saliva that can be obtained by non-invasive methods, have been appeared [7, 8]. It has been studied that under bronchial asthma considerable changes in exhaled breath condensate appeared , including its physical and chemical properties , acid-base balance, accumulation both nitric oxide and hydrogen peroxide, as well as aldehydes, prostanoids and cytokines [5, 9, 10]. However, most studies of exhaled breath condensate have been carried out in adults, but in children are almost absent, which assign the topicality of this research, aimed at study of inflammatory processes and improvement of individualized treatment for children with different phenotypes of bronchial asthma. "
    [Show abstract] [Hide abstract] ABSTRACT: Summary. The indices of exhaled air condensate have been studied in 160 children with different phenotypes of bronchial asthma. It has been demonstrated that there occur changes in children with the noneosinophilic phenotype of bronchial asthma as compared with the patients with the eosinophilic phenotype of the disease that are indicative of a higher activity of inflammatory processes in the respiratory tracts. The proteolytic activity according to azocasein lysis more than 1,5 ml/h had the highest diagnostic value of detection of noneosinophilic phenotype: spesiphity 84,7 %, the predictable value of a positive result 79,1 %, the odds ratio 7,5, the posttesting probability of positive result 79,1 %. Key words: children, bronchial asthma, exhaled breath condensate.
    Full-text · Article · Jun 2013 · Annals of agricultural and environmental medicine: AAEM
Show more