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1.
Microbial volatiles as diagnostic biomarkers of bacterial lung infection in mechanically ventilated patients
Waqar M Ahmed, Dominic Fenn, Iain R. White, Breanna Dixon, Tamara M E Nijsen, Hugo H Knobel, Paul Brinkman, Pouline M P van Oort, Marcus J Schultz, Paul Dark, Royston Goodacre, Timothy Felton, Lieuwe D J Bos, Stephen J Fowler, 2022, original scientific article

Abstract: Background Early and accurate recognition of respiratory pathogens is crucial to prevent increased risk of mortality in critically ill patients. Microbial-derived volatile organic compounds (mVOCs) in exhaled breath could be used as non-invasive biomarkers of infection to support clinical diagnosis. Methods In this study, we investigated the diagnostic potential of in vitro confirmed mVOCs in the exhaled breath of patients under mechanically ventilation from the BreathDx study. Samples were analysed by thermal desorption-gas chromatography-mass spectrometry (TD-GC-MS). Results Pathogens from bronchoalveolar lavage (BAL) cultures were identified in 45/89 patients and S. aureus was the most commonly identified pathogen (n = 15). Out of 19 mVOCs detected in the in vitro culture headspace of four common respiratory pathogens (Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae and Escherichia coli), 14 were found in exhaled breath samples. Higher concentrations of two mVOCs were found in the exhaled breath of patients infected with S. aureus compared to those without (3-methylbutanal p < 0.01. AUROC = 0.81-0.87 and 3-methylbutanoic acid p = 0.01. AUROC = 0.79-0.80). In addition, bacteria identified from BAL cultures which are known to metabolise tryptophan (Escherichia coli, Klebsiella oxytoca and Haemophilus influenzae) were grouped and found to produce higher concentrations of indole compared to breath samples with culture-negative (p = 0.034) and other pathogen-positive (p = 0.049) samples. Conclusions This study demonstrates the capability of using mVOCs to detect the presence of specific pathogen groups with potential to support clinical diagnosis. Although not all mVOCs were found in patient samples within this small pilot study, further targeted and qualitative investigation is warranted using multi-centre clinical studies.
Keywords: Breath, VOCs, infection, respiratory pathogens, VAP
Published in RUNG: 28.11.2022; Views: 1164; Downloads: 0
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2.
Breath and plasma metabolomics to assess inflammation in acute stroke
Waqar Ahmed, Iain R. White, Maxim Wilkinson, Craig Johnson, Nicholas J. W. Rattray, Amit K. Kishore, Royston Goodacre, Craig J. Smith, Stephen J. Fowler, 2021, original scientific article

Abstract: Inflammation is strongly implicated in both injury and repair processes occurring after stroke. In this exploratory study we assessed the feasibility of repeated sampling of exhaled volatile organic compounds and performed an untargeted metabolomic analysis of plasma collected at multiple time periods after stroke. Metabolic profiles were compared with the time course of the inflammatory markers C-reactive protein (CRP) and interleukin-6 (IL-6). Serial breath sampling was well-tolerated by all patients and the measurement appears feasible in this group. We found that exhaled decanal tracks CRP and IL-6 levels post-stroke and correlates with several metabolic pathways associated with a post-stroke inflammatory response. This suggests that measurement of breath and blood metabolites could facilitate development of novel therapeutic and diagnostic strategies. Results are discussed in relation to the utility of breath analysis in stroke care, such as in monitoring recovery and complications including stroke associated infection.
Keywords: stroke, metabolomics, breath, VOCs, inflammation
Published in RUNG: 18.11.2021; Views: 1621; Downloads: 62
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3.
Effects of high relative humidity and dry purging on VOCs obtained during breath sampling on common sorbent tubes
Maxim Wilkinson, Iain R. White, Roy Goodacre, Tamara Nijsen, Stephen Fowler, 2020, original scientific article

Abstract: Offline breath analysis by thermal desorption gas chromatography mass spectrometry (TD-GC-MS) requires the use of sorbent traps to concentrate and store volatile compounds. The selection of which sorbent to use and best practices for managing high relative humidity are important considerations to allow for reproducible, untargeted, biomarker discovery in water saturated breath samples. This work aims to assess three commonly used sorbent materials for their use in breath volatile sampling and determine how the high relative humidity inherent in such samples effects the capture of volatile compounds of interest. TenaxGR, TenaxTA/Carbograph1TD and TenaxTA/Carbograph5TD tubes were selected as they are the most commonly used sorbents in the breath sampling literature. The recovery of 29 compounds in a standard mix loaded using high humidity gas was tested for each sorbent and compared to loading in dry gas. Water retention and dry purge rates were determined for each sorbent for 500 ml and 1000 ml breath collections. Finally, breath samples were collected simultaneously on to each sorbent type using the ReCIVA and analysed by TD-GC-MS. All three sorbents exhibited acceptable reproducibility when loaded with the standard mix in dry gas (RSD < 10%). Loading the standard mix in humid gas led to reduced recovery of compounds based on their chemical properties. Dry purging performance for each sorbent material was assessed and was shown to be 1.14, 1.13 and 0.89 mg H2O min−1 for TenaxGR, TenaxTA/Carbograph1TD and TenaxTA/Carbograph5TD respectively when flushed with 50 ml min−1 of N2. A comparison of breath profiles on different sorbents showed differences in background artefacts (sulfur dioxide, cyclopenten-1-one and 3-nonene) and endogenous breath compounds (2-methyl-furan and furfural). This work demonstrates that high relative humidity during sampling reduces the ability of sorbent tubes to capture volatile compounds and could impact method detection limits during breath sampling. Sufficient water to impair accurate analysis was retained on all tubes. Minimal differences were observed between sorbent materials when used to sample breath, however, suggestions are provided for sorbent selection for future studies.
Keywords: VOCs, Breath sampling, ReCIVA
Published in RUNG: 27.07.2020; Views: 2832; Downloads: 108
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4.
Under fungal attack on a metalliferous soil: ROS or not ROS? Insights from Silene paradoxa L. growing under copper stress
Cosimo Taiti, Elisabetta Giorni, Ilaria Colzi, Sara Pignattelli, Nadia Bazihizina, Antonella Buccianti, Simone Luti, Luigia Pazzagli, Stefano Mancuso, Cristina Gonnelli, 2016, original scientific article

Abstract: We investigated how the adaptation to metalliferous environments can influence the plant response to biotic stress. In a metallicolous and a non-metallicolous population of Silene paradoxa the induction of oxidative stress and the production of callose and volatiles were evaluated in the presence of copper and of the PAMP fungal protein cerato-platanin, separately and in combination. Our results showed incompatibility between the ordinary ROS-mediated response to fungal attack and the acquired mechanisms of preventing oxidative stress in the tolerant population. A similar situation was also demonstrated by the sensitive population growing in the presence of copper but, in this case, with a lack of certain responses, such as callose production. In addition, in terms of the joint behaviour of emitted volatiles, multivariate statistics showed that not only did the populations respond differently to the presence of copper or biotic stress, but also that the biotic and abiotic stresses interacted in different ways in the two populations. Our results demonstrated that the same incompatibility of hyperaccumulators in ROS-mediated biotic stress signals also seemed to be exhibited by the excluder metallophyte, but without the advantage of being able to rely on the elemental defence for plant protection from natural enemies.
Keywords: Biotic interactions Callose Heavy metals Oxidative stress VOCs
Published in RUNG: 20.04.2020; Views: 3297; Downloads: 0
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5.
A microbiome and metabolomic signature of phases of cutaneous healing identified by profiling sequential acute wounds of human skin: An exploratory study
Mohammed Ashrafi, Yun Xu, Howbeer Muhamadali, Iain R. White, Maxim Wilkinson, Mohamed Baguneid, Roy Goodacre, Ardeshir Bayat, 2020, original scientific article

Abstract: Profiling skin microbiome and metabolome has been utilised to gain further insight into wound healing processes. The aims of this multi-part temporal study in 11 volunteers were to analytically profile the dynamic wound tissue and headspace metabolome and sequence microbial communities in acute wound healing at days 0, 7, 14, 21 and 28, and to investigate their relationship to wound healing, using non-invasive quantitative devices. Metabolites were obtained using tissue extraction, sorbent and polydimethylsiloxane patches and analysed using GCMS. PCA of wound tissue metabolome clearly separated time points with 10 metabolites of 346 being involved in separation. Analysis of variance-simultaneous component analysis identified a statistical difference between the wound headspace metabolome, sites (P = 0.0024) and time points (P<0.0001), with 10 out of the 129 metabolites measured involved with this separation between sites and time points. A reciprocal relationship between Staphylococcus spp. and Propionibacterium spp. was observed at day 21 (P<0.05) with a statistical correlation between collagen and Propionibacterium (r = 0.417; P = 0.038) and Staphylococcus (r = -0.434; P = 0.03). Procrustes analysis showed a statistically significant similarity between wound headspace and tissue metabolome with non-invasive wound devices. This exploratory study demonstrates the temporal and dynamic nature of acute wound metabolome and microbiome presenting a novel class of biomarkers that correspond to wound healing, with further confirmatory studies now necessary.
Keywords: metabolomics, skin, volatile organic compounds, VOCs, wound healing
Published in RUNG: 03.03.2020; Views: 2943; Downloads: 0
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6.
Circadian rhythm of exhaled biomarkers in health and asthma
Max Wilkinson, Robert Maidstone, Andrew Loudon, John Blaikley, Iain R. White, Dave Singh, David Ray, Royston Goodacre, Stephen Fowler, Hannah Durrington, 2019, original scientific article

Abstract: Circadian rhythms control many biological processes in the body in both health and disease. Greater understanding of diurnal variability in disease related biomarkers is crucial for their application in clinical practice and biomarkers of circadian rhythm are required to facilitate further research into disturbed chronicity. To determine if fractional exhaled nitric oxide and breath volatile biomarkers vary rhythmically during the day in healthy and asthmatic individuals. Ten individuals with moderate, atopic asthma (on regular inhaled corticosteroids) and 10 healthy volunteers (all non-smokers) completed an overnight visit where their exhaled breath volatiles and forced exhaled nitric oxide levels were collected every 6 h. Breath volatiles were analysed using gas chromatography mass spectrometry, after trapping these volatiles on sorbent materials for thermal desorption. Nine breath volatiles (including acetone and isoprene) exhibit diurnal variation across all individuals. Furthermore the circadian pattern of several VOCs is altered in individuals with asthma and fractional exhaled nitric oxide is rhythmic in asthma but not in healthy controls. Markers of circadian rhythm can be identified in breath and may offer insight into circadian profiling to help treat disease. Additionally this work suggests that time of day must be controlled when designing future biomarker discovery studies. Further work is required with larger cohorts to validate and extend these findings.
Keywords: VOCs, breath, asthma, circadian
Published in RUNG: 21.10.2019; Views: 3182; Downloads: 0
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7.
Breathomics and its Application for Disease Diagnosis: A Review of Analytical Techniques and Approaches
David J Beale, Oliver A H Jones, Avinash V Karpe, Ding Y Oh, Iain R. White, Konstantinos A Kouremenos, Enzo A Palombo, 2018, independent scientific component part or a chapter in a monograph

Abstract: The application of metabolomics to an ever-greater variety of sample types is a key focus of systems biology research. Recently, there has been a strong focus on applying these approaches toward the rapid analysis of metabolites found in non-invasively acquired samples, such as exhaled breath (also known as ‘breathomics’). The sampling process involved in collecting exhaled breath is nonintrusive and comparatively low-cost. It uses a series of globally approved methods and provides researchers with easy access to the metabolites secreted by the human body. Owing to its accuracy and rapid nature, metabolomic analysis of breath is a rapidly growing field that has proven effective in detecting and diagnosing the early stages of numerous diseases and infections. This review discusses the various collection and analysis methods currently applied in breathomics research. Some of the salient research completed in this field to date is also assessed and discussed in order to provide a basis for possible future scientific directions.
Keywords: Metabolomics, breath research, VOCs, breathomics
Published in RUNG: 22.07.2019; Views: 3064; Downloads: 0
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8.
Metabolite profiling of the ripening of Mangoes Mangifera indica L. cv. ‘Tommy Atkins’ by real-time measurement of volatile organic compounds
Iain R. White, Robert S Blake, Andrew J Taylor, Paul S Monks, 2016, original scientific article

Abstract: Real-time profiling of mango ripening based on proton transfer reaction-time of flight-mass spectrometry (PTR–ToF–MS) of small molecular weight volatile organic compounds (VOCs), is demonstrated using headspace measurements of ‘Tommy Atkins’ mangoes. VOC metabolites produced during the ripening process were sampled directly, which enabled simultaneous and rapid detection of a wide range of compounds. Headspace measurements of ‘Keitt’ mangoes were also conducted for comparison. A principle component analysis of the results indicated that several mass channels were not only key to the ripening process but could also be used to distinguish between mango cultivars. The identities of 22 of these channels, tentatively speciated using contemporaneous GC–MS measurements of sorbent tubes, are rationalized through examination of the biochemical pathways that produce volatile flavour components. Results are discussed with relevance to the potential of headspace analysers and electronic noses in future fruit ripening and quality studies.
Keywords: Mangifera indica, Tommy Atkins, PTR–ToF–MS, VOCs, Ripening, Mango
Published in RUNG: 18.07.2019; Views: 2846; Downloads: 0
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9.
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