breath condensate
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Molecules ◽  
2022 ◽  
Vol 27 (2) ◽  
pp. 549
Mariana Santos-Rivera ◽  
Amelia R. Woolums ◽  
Merrilee Thoresen ◽  
Florencia Meyer ◽  
Carrie K. Vance

Bovine respiratory syncytial virus (BRSV) is a major contributor to respiratory disease in cattle worldwide. Traditionally, BRSV infection is detected based on non-specific clinical signs, followed by reverse transcriptase-polymerase chain reaction (RT-PCR), the results of which can take days to obtain. Near-infrared aquaphotomics evaluation based on biochemical information from biofluids has the potential to support the rapid identification of BRSV infection in the field. This study evaluated NIR spectra (n = 240) of exhaled breath condensate (EBC) from dairy calves (n = 5) undergoing a controlled infection with BRSV. Changes in the organization of the aqueous phase of EBC during the baseline (pre-infection) and infected (post-infection and clinically abnormal) stages were found in the WAMACS (water matrix coordinates) C1, C5, C9, and C11, likely associated with volatile and non-volatile compounds in EBC. The discrimination of these chemical profiles by PCA-LDA models differentiated samples collected during the baseline and infected stages with an accuracy, sensitivity, and specificity >93% in both the calibration and validation. Thus, biochemical changes occurring during BRSV infection can be detected and evaluated with NIR-aquaphotomics in EBC. These findings form the foundation for developing an innovative, non-invasive, and in-field diagnostic tool to identify BRSV infection in cattle.

2022 ◽  
Vol 11 (1) ◽  
pp. 252
Joanna Połomska ◽  
Barbara Sozańska

(1) Background: L-arginine (L-ARG) and its metabolites are involved in some aspects of asthma pathogenesis (airway inflammation, oxidative stress, bronchial responsiveness, collagen deposition). Published data indicate that lungs are a critical organ for the regulation of L-ARG metabolism and that alterations in L-ARG metabolism may be significant for asthma. The aim of this study was to assess the levels of L-ARG and its metabolites in pediatric patients with asthma in serum and exhaled breath condensate (EBC) by mass spectrometric analysis and compare them with non-asthmatic children. (2) Methods: Sixty-five children (37 pediatric patients with bronchial asthma and 28 healthy control subjects) aged 6–17 participated in the study. All participants underwent a clinical visit, lung tests, allergy tests with common aeroallergens, and serum and EBC collection. The levels of biomarkers were determined in both serum and EBC. Analytical chromatography was conducted using an Acquity UPLC system equipped with a cooled autosampler and an Acquity HSS T3 column. Mass spectrometric analysis was conducted using the Xevo G2 QTOF MS with electrospray ionization (ESI) in positive ion mode. (3) Results: Asymmetric dimethylarginine (ADMA) and symmetric dimethylarginine (SDMA) levels in serum and EBC did not differ significantly in asthmatic children and healthy control subjects. We found no correlation between forced expiratory volume in one second (FEV1) and L-ARG and its metabolites, as well as between interleukin-4 (IL-4) serum level and L-ARG and its metabolites. Concentrations of ADMA, SDMA, citrulline (CIT), and ornithine (ORN) were higher in serum than EBC in asthmatics and non-asthmatics. By contrast, concentrations of dimethylarginine (DMA) were higher in EBC than serum. ADMA/L-ARG, SDMA/L-ARG, and DMA/L-ARG ratios were significantly higher in EBC than in serum in asthmatics and in non-asthmatics. (4) Conclusions: Serum and EBC concentrations of L-ARG and its metabolites were not an indicator of pediatric bronchial asthma in our study.

Amr Kamel Khalil Ahmed ◽  
Mahmoud El Kazzaz

COVID-19 has emerged as a global pandemic. It mainly manifests as pneumonia, which may deteriorate into severe respiratory failure. The major hallmark of the disease is the systemic inflammatory immune response characterized by cytokine storm (CS). CS is marked by elevated levels of inflammatory cytokines, mainly interleukin-6 (IL-6), IL-8, IL-10, tumor necrosis factor-α (TNF-α) and interferon-γ (IFNγ). Of these, IL-6 is found to be significantly associated with higher mortality. IL-6 is also a robust marker for predicting disease prognosis and deterioration of the clinical profile. (1) IL-6 was detectable in the breath condensate of all healthy nonsmokers but was significantly higher in COPD patients. Exhaled breath condensate is totally noninvasive and highly acceptable to patients. The collection procedure has no effect on airway function or inflammation, and there is growing evidence that abnormalities in condensate composition may reflect biochemical changes in airway lining fluid. This method has been successfully used in previous studies to investigate several inflammatory markers in COPD and asthmatic patients. (2) Il-6 is produced in the lung by interstitial fibroblasts, alveolar macrophages, and large-vessel and bronchial epithelial cells.These studies and our novel method after clinical trials may open the field for future therapies for COVID-19 and post-COVID-19 lung fibrosis by inhaler transport medicines as a new challenge for overcoming sequelae of this pandemic. This suggested a new procedure for the measurement of exhaled IL-6. We studied which IL-6 is risky is IL-6 produced from airway or endocrine IL-6 or immune IL-6. We can classify IL-6 as cytokine storm under pathological conditions to three mains of sources as immune IL-6 produced from respiratory system like interstitial fibroblasts and bronchial epithelial cells and fibroblast, second endocrine IL-6 produced from zona glomerulosa of adrenal glands stimulated by ACTH and immune IL-6 from macrophages and other immune cells. From previous studies we need a procedure acceptable to patients and easy, noninvasive, sensitive so we plan for clinical trial registered at about measurement of interleukin-6 at exhaled condensate of covid-19 patients and post-19 covid patients with lung fibrosis as a novel study which may be a useful tool, easy, sensitive for early intervention with anti-il6 medications and determine the degree of severity by inflammatory markers for intensity of airway inflammation.

S Riscassi ◽  
M Corradi ◽  
R Andreoli ◽  
C Maccari ◽  
F Mercolini ◽  

Metabolites ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 847
Elettra Barberis ◽  
Elia Amede ◽  
Shahzaib Khoso ◽  
Luigi Castello ◽  
Pier Paolo Sainaghi ◽  

Infection from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can lead to severe respiratory tract damage and acute lung injury. Therefore, it is crucial to study breath-associated biofluids not only to investigate the breath’s biochemical changes caused by SARS-CoV-2 infection, but also to discover potential biomarkers for the development of new diagnostic tools. In the present study, we performed an untargeted metabolomics approach using a bidimensional gas chromatography mass spectrometer (GCxGC-TOFMS) on exhaled breath condensate (EBC) from COVID-19 patients and negative healthy subjects to identify new potential biomarkers for the noninvasive diagnosis and monitoring of the COVID-19 disease. The EBC analysis was further performed in patients with acute or acute-on-chronic cardiopulmonary edema (CPE) to assess the reliability of the identified biomarkers. Our findings demonstrated that an abundance of EBC fatty acids can be used to discriminate COVID-19 patients and that they may have a protective effect, thus suggesting their potential use as a preventive strategy against the infection.

Shuang Hu ◽  
Mitchell M McCartney ◽  
Juan Arredondo ◽  
Sumathi Sankaran-Walters ◽  
Eva Borras ◽  

Abstract Exhaled breath condensate (EBC) is routinely collected and analyzed in breath research. Because it contains aerosol droplets, EBC samples from SARS-CoV-2 infected individuals harbor the virus and pose the threat of infectious exposure. We report for the first time a safe and consistent method to fully inactivate SARS-CoV-2 in EBC samples and make EBC samples safe for processing and analysis. EBC samples containing infectious SARS-CoV-2 were treated with several concentrations of acetonitrile. The most commonly used 10% acetonitrile treatment for EBC processing failed to completely inactivate the virus in samples and viable virus was detected by the assay of SARS-CoV-2 infection of Vero E6 cells in a BSL-3 laboratory. Treatment with either 50% or 90% acetonitrile was effective to completely inactivate the virus, resulting in safe, non-infectious EBC samples that can be used for metabolomic analysis. Our study provides SARS-CoV-2 inactivation protocol for the collection and processing of EBC samples in the clinical setting and for advancing to metabolic assessments in health and disease.

2021 ◽  
Parisa Eydi ◽  
Elaheh Rahimpour ◽  
Maryam Khoubnasabjafari ◽  
Vahid Jouyban-Gharamaleki ◽  
Abolghasem Jouyban

Background: Lamotrigine is widely used in the management of partial epilepsy, generalized tonic-clonic epilepsy and Lenox-Gastut syndrome and an add-on therapy in the treatment of complex and simple partial seizures and secondarily generalized tonic-clonic seizures resistant to multiple drug therapy. Methods: In the current study, a fluorometric nanoprobe based on metal–organic frameworks (MOF) was designed for the determination of lamotrigine in exhaled breath condensate (EBC). The MOF nanoprobe consisted of Tb3+ ions as metal part and dimethylformamide (DMF) and 1,10-phenanthroline (Phen) as organic parts of nanoprobe. Results: The used probe shows a week fluorescence in alkaline media owing to an energy transfer from nitrogen groups of DMF and Phen on carbonyl group of DMF as an antenna for Tb3+ luminescence. However, its fluorescence is enhanced in acidic conditions by protonation of DMF nitrogen atoms and Phen and deactivation of energy transfer pathways of nitrogen groups to carbonyl group. Lamotrigine addition to this fluorescent system leads to quenching in the fluorescence intensity due to reactivation of the above mentioned energy transfer pathways resulting in competitive interaction with H+ ions. Moreover, the inner filter effect (IFE) of lamotrigine on DMF–Tb–Phen MOF NPs is considered as another reason for the observed quenching in the fluorescence of DMF–Tb–Phen MOF NPs. The intensity of the fluorescence was recorded at λem = 545 nm and the difference between fluorescence signal in the absence and presence of lamotrigine was the analytical response. The factors affected on experimental conditions were optimized utilizing a multivariate optimization technique. The validation of nanoprobe response to lamotrigine gives a linear relationship in the range of 0.05 to 2.0 µg⋅mL‒1 with a detection limit of 11.0 ng⋅mL‒1 for lamotrigine. Conclusion: The developed method reveals good repeatability and selectivity for lamotrigine in real samples.

2021 ◽  
Vol 10 (21) ◽  
pp. 5165
Inger Lise Gade ◽  
Jacob Gammelgaard Schultz ◽  
Rasmus Froberg Brøndum ◽  
Benedict Kjærgaard ◽  
Jens Erik Nielsen-Kudsk ◽  

Current diagnostic markers for pulmonary embolism (PE) are unspecific. We investigated the proteome of the exhaled breath condensate (EBC) in a porcine model of acute PE in order to identify putative diagnostic markers for PE. EBC was collected at baseline and after the induction of autologous intermediate-risk PE in 14 pigs, plus four negative control pigs. The protein profiles of the EBC were analyzed using label-free quantitative nano liquid chromatography–tandem mass spectrometry. A total of 897 proteins were identified in the EBCs from the pigs. Alterations were found in the levels of 145 different proteins after PE compared with the baseline and negative controls: albumin was among the most upregulated proteins, with 14-fold higher levels 2.5 h after PE (p-value: 0.02). The levels of 49 other proteins were between 1.3- and 17.1-fold higher after PE. The levels of 95 proteins were lower after PE. Neutrophil gelatinase-associated lipocalin (fold change 0.3, p-value < 0.01) was among the most reduced proteins 2.5 h after PE. A prediction model based on penalized regression identified five proteins including albumin and neutrophil gelatinase-associated lipocalin. The model was capable of discriminating baseline samples from EBC samples collected 2.5 h after PE correctly in 22 out of 27 samples. In conclusion, the EBC from pigs with acute PE contained several putative diagnostic markers of PE.

2021 ◽  
pp. 212-223
S. Yu. Tereshchenko ◽  
M. A. Malinchik ◽  
M. V. Smolnikova

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.

Antioxidants ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 1661
Maud Hemmendinger ◽  
Jean-Jacques Sauvain ◽  
Nancy B. Hopf ◽  
Pascal Wild ◽  
Guillaume Suárez ◽  

There are several methods for quantifying malondialdehyde (MDA), an oxidative stress biomarker, in exhaled breath condensate (EBC). However, due to the very diluted nature of this biological matrix, a high variability is observed at low concentrations. We aimed to optimize a 2,4-dinitrophenylhydrazine-based method using liquid chromatography coupled to tandem mass spectrometry and characterize the uncertainty associated with this method. We investigated the following parameters for the method validation: calibration linearity, limit of detection (LOD), precision, recovery, and matrix effect. The results were used to identify the main sources of uncertainty and calculating the combined uncertainty. The applicability of this method was evaluated in an ongoing epidemiological study by analyzing 164 EBC samples collected from different professional groups in subway environments. The optimized method was sensitive (LOD: 70 pg/mL), precise (inter-day variation < 19%) and accurate (recovery range: 92–106.5%). The calculated analytical uncertainty was the highest at the LOQ level and reached 23%. Although the analytical uncertainty was high at low MDA concentrations, it was significantly lower than that the observed inter-individual variability. Hence, this method performs sufficiently well and can be recommended for future use in epidemiological researches relying on between-subject differences.

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