Drivers of natural gas use in U.S. residential buildings.

Natural gas is the primary fuel used in U.S. residences, yet little is known about its consumption patterns and drivers. We use daily county-level gas consumption data to assess the spatial patterns of the relationships and the sensitivities of gas consumption to outdoor air temperature across U.S. households. We fitted linear-plus-plateau functions to daily gas consumption data in 1000 counties, and derived two key coefficients: the heating temperature threshold (Tcrit) and the gas consumption rate change per 1°C temperature drop (Slope). We identified the main predictors of Tcrit and Slope (like income, employment rate, and building type) using interpretable machine learning models built on census data. Finally, we estimated a potential 2.47 million MtCO2 annual emission reduction in U.S. residences by gas savings due to household insulation improvements and hypothetical behavioral change toward reduced consumption by adopting a 1°C lower Tcrit than the current value.

Complex adaptive systems-based framework for modeling the health impacts of climate change.

Introduction: Climate change is a global phenomenon with far-reaching consequences, and its impact on human health is a growing concern. The intricate interplay of various factors makes it challenging to accurately predict and understand the implications of climate change on human well-being. Conventional methodologies have limitations in comprehensively addressing the complexity and nonlinearity inherent in the relationships between climate change and health outcomes. Objectives: The primary objective of this paper is to develop a robust theoretical framework that can effectively analyze and interpret the intricate web of variables influencing the human health impacts of climate change. By doing so, we aim to overcome the limitations of conventional approaches and provide a more nuanced understanding of the complex relationships involved. Furthermore, we seek to explore practical applications of this theoretical framework to enhance our ability to predict, mitigate, and adapt to the diverse health challenges posed by a changing climate. Methods: Addressing the challenges outlined in the objectives, this study introduces the Complex Adaptive Systems (CAS) framework, acknowledging its significance in capturing the nuanced dynamics of health effects linked to climate change. The research utilizes a blend of field observations, expert interviews, key informant interviews, and an extensive literature review to shape the development of the CAS framework. Results and discussion: The proposed CAS framework categorizes findings into six key sub-systems: ecological services, extreme weather, infectious diseases, food security, disaster risk management, and clinical public health. The study employs agent-based modeling, using causal loop diagrams (CLDs) tailored for each CAS sub-system. A set of identified variables is incorporated into predictive modeling to enhance the understanding of health outcomes within the CAS framework. Through a combination of theoretical development and practical application, this paper aspires to contribute valuable insights to the interdisciplinary field of climate change and health. Integrating agent-based modeling and CLDs enhances the predictive capabilities required for effective health outcome analysis in the context of climate change. Conclusion: This paper serves as a valuable resource for policymakers, researchers, and public health professionals by employing a CAS framework to understand and assess the complex network of health impacts associated with climate change. It offers insights into effective strategies for safeguarding human health amidst current and future climate challenges.

Effects of Contagious Respiratory Pathogens on Breath Biomarkers.

Due to their immediate exhalation after generation at the cellular/microbiome levels, exhaled volatile organic compounds (VOCs) may provide real-time information on pathophysiological mechanisms and the host response to infection. In recent years, the metabolic profiling of the most frequent respiratory infections has gained interest as it holds potential for the early, non-invasive detection of pathogens and the monitoring of disease progression and the response to therapy. Using previously unpublished data, randomly selected individuals from a COVID-19 test center were included in the study. Based on multiplex PCR results (non-SARS-CoV-2 respiratory pathogens), the breath profiles of 479 subjects with the presence or absence of flu-like symptoms were obtained using proton-transfer-reaction time-of-flight mass spectrometry. Among 223 individuals, one respiratory pathogen was detected in 171 cases, and more than one pathogen in 52 cases. A total of 256 subjects had negative PCR test results and had no symptoms. The exhaled VOC profiles were affected by the presence of Haemophilus influenzae, Streptococcus pneumoniae, and Rhinovirus. The endogenous ketone, short-chain fatty acid, organosulfur, aldehyde, and terpene concentrations changed, but only a few compounds exhibited concentration changes above inter-individual physiological variations. Based on the VOC origins, the observed concentration changes may be attributed to oxidative stress and antioxidative defense, energy metabolism, systemic microbial immune homeostasis, and inflammation. In contrast to previous studies with pre-selected patient groups, the results of this study demonstrate the broad inter-individual variations in VOC profiles in real-life screening conditions. As no unique infection markers exist, only concentration changes clearly above the mentioned variations can be regarded as indicative of infection or colonization.

Protocol for visualizing complex volatile metabolomics data in clinical setups using EDaViS software.

Here, we present a protocol for using Early Data Visualization Script, a user-friendly software tool to visualize complex volatile metabolomics data in clinical setups. We describe steps for tabulating data and adjusting visual output to visualize complex time-resolved volatile omics data using simple charts and graphs. We then demonstrate possible modifications by detailing procedures for the adaptation of four basic functions. For complete details on the use and execution of this protocol, please refer to Sukul et al. (2022)1 and Remy et al. (2022).2.

Origin of breath isoprene in humans is revealed via multi-omic investigations.

Plants, animals and humans metabolically produce volatile isoprene (C5H8). Humans continuously exhale isoprene and exhaled concentrations differ under various physio-metabolic and pathophysiological conditions. Yet unknown metabolic origin hinders isoprene to reach clinical practice as a biomarker. Screening 2000 individuals from consecutive mass-spectrometric studies, we herein identify five healthy German adults without exhaled isoprene. Whole exome sequencing in these adults reveals only one shared homozygous (European prevalence: <1%) IDI2 stop-gain mutation, which causes losses of enzyme active site and Mg2+-cofactor binding sites. Consequently, the conversion of isopentenyl diphosphate to dimethylallyl diphosphate (DMAPP) as part of the cholesterol metabolism is prevented in these adults. Targeted sequencing depicts that the IDI2 rs1044261 variant (p.Trp144Stop) is heterozygous in isoprene deficient blood-relatives and absent in unrelated isoprene normal adults. Wild-type IDI1 and cholesterol metabolism related serological parameters are normal in all adults. IDI2 determines isoprene production as only DMAPP sources isoprene and unlike plants, humans lack isoprene synthase and its enzyme homologue. Human IDI2 is expressed only in skeletal-myocellular peroxisomes and instant spikes in isoprene exhalation during muscle activity underpins its origin from muscular lipolytic cholesterol metabolism. Our findings translate isoprene as a clinically interpretable breath biomarker towards potential applications in human medicine.

Advanced setup for safe breath sampling and patient monitoring under highly infectious conditions in the clinical environment.

Being the proximal matrix, breath offers immediate metabolic outlook of respiratory infections. However, high viral load in exhalations imposes higher transmission risk that needs improved methods for safe and repeatable analysis. Here, we have advanced the state-of-the-art methods for real-time and offline mass-spectrometry based analysis of exhaled volatile organic compounds (VOCs) under SARS-CoV-2 and/or similar respiratory conditions. To reduce infection risk, the general experimental setups for direct and offline breath sampling are modified. Certain mainstream and side-stream viral filters are examined for direct and lab-based applications. Confounders/contributions from filters and optimum operational conditions are assessed. We observed immediate effects of infection safety mandates on breath biomarker profiles. Main-stream filters induced physiological and analytical effects. Side-stream filters caused only systematic analytical effects. Observed substance specific effects partly depended on compound's origin and properties, sampling flow and respiratory rate. For offline samples, storage time, -conditions and -temperature were crucial. Our methods provided repeatable conditions for point-of-care and lab-based breath analysis with low risk of disease transmission. Besides breath VOCs profiling in spontaneously breathing subjects at the screening scenario of COVID-19/similar test centres, our methods and protocols are applicable for moderately/severely ill (even mechanically-ventilated) and highly contagious patients at the intensive care.

Profiling of exhaled volatile organics in the screening scenario of a COVID-19 test center.

Breath volatile organics (VOCs) may provide immediate information on infection mechanisms and host response. We conducted real-time mass spectrometry-based breath profiling in 708 non-preselected consecutive subjects in the screening scenario of a COVID-19 test center. Recruited subjects were grouped based on PCR-confirmed infection status and presence or absence of flu-like symptoms. Exhaled VOC profiles of SARS-CoV-2-positive cases (n = 36) differed from healthy (n = 256) and those with other respiratory infections (n = 416). Concentrations of most VOCs were suppressed in COVID-19. VOC concentrations also differed between symptomatic and asymptomatic cases. Breath markers mirror effects of infections onto host's cellular metabolism and microbiome. Downregulation of specific VOCs was attributed to suppressive effects of SARS-CoV-2 onto gut or pulmonary microbial metabolism. Breath analysis holds potential for monitoring SARS-CoV-2 infections rather than for primary diagnosis. Breath profiling offers unconventional insight into host-virus cross-talk and infection microbiology and enables non-invasive assessment of disease manifestation.

Real-time metabolic monitoring under exhaustive exercise and evaluation of ventilatory threshold by breathomics: Independent validation of evidence and advances.

Breath analysis was coupled with ergo-spirometry for non-invasive profiling of physio-metabolic status under exhaustive exercise. Real-time mass-spectrometry based continuous analysis of exhaled metabolites along with breath-resolved spirometry and heart rate monitoring were executed while 14 healthy adults performed ergometric ramp exercise protocol until exhaustion. Arterial blood lactate level was analyzed at defined time points. Respiratory-cardiac parameters and exhalation of several blood-borne volatiles changed continuously with the course of exercise and increasing workloads. Exhaled volatiles mirrored ventilatory and/or hemodynamic effects and depended on the origin and/or physicochemical properties of the substances. At the maximum workload, endogenous isoprene, methanethiol, dimethylsulfide, acetaldehyde, butanal, butyric acid and acetone concentrations decreased significantly by 74, 25, 35, 46, 21, 2 and 2%, respectively. Observed trends in exogenous cyclohexadiene and acetonitrile mimicked isoprene profile due to their similar solubility and volatility. Assignment of anaerobic threshold was possible via breath acetone. Breathomics enabled instant profiling of physio-metabolic effects and anaerobic thresholds during exercise. Profiles of exhaled volatiles indicated effects from muscular vasoconstriction, compartmental distribution of perfusion, extra-alveolar gas-exchange and energy homeostasis. Sulfur containing compounds and butyric acid turned out to be interesting for investigations of combined diet and exercise programs. Reproducible metabolic breath patterns have enhanced scopes of breathomics in sports science/medicine.

Non-Invasive O-Toluidine Monitoring during Regional Anaesthesia with Prilocaine and Detection of Accidental Intravenous Injection in an Animal Model.

Regional anaesthesia is well established as a standard method in clinical practice. Currently, the local anaesthetics of amino-amide types such as prilocaine are frequently used. Despite routine use, complications due to overdose or accidental intravenous injection can arise. A non-invasive method that can indicate such complications early would be desirable. Breath gas analysis offers great potential for the non-invasive monitoring of drugs and their volatile metabolites. The physicochemical properties of o-toluidine, the main metabolite of prilocaine, allow its detection in breath gas. Within this study, we investigated whether o-toluidine can be monitored in exhaled breath during regional anaesthesia in an animal model, if correlations between o-toluidine and prilocaine blood levels exist and if accidental intravenous injections are detectable by o-toluidine breath monitoring. Continuous o-toluidine monitoring was possible during regional anaesthesia of the cervical plexus and during simulated accidental intravenous injection of prilocaine. The time course of exhaled o-toluidine concentrations considerably differed depending on the injection site. Intravenous injection led to an immediate increase in exhaled o-toluidine concentrations within 2 min, earlier peak and higher maximum concentrations, followed by a faster decay compared to regional anaesthesia. The strength of correlation of blood and breath parameters depended on the injection site. In conclusion, real time monitoring of o-toluidine in breath gas is possible by means of PTR-ToF-MS. Since simulated accidental intravenous injection led to an immediate increase in exhaled o-toluidine concentrations within 2 min and higher maximum concentrations, monitoring exhaled o-toluidine may potentially be applied for the non-invasive real-time detection of accidental intravenous injection of prilocaine.

Physiological and metabolic effects of healthy female aging on exhaled breath biomarkers.

Healthy aging driven physio-metabolic events in females hold the key to complex in vivo mechanistic links and systemic cross talks. Effects from basic changes at genome, proteome, metabolome, and lipidome levels are often reflected at the upstream phenome (e.g., breath volatome) cascades. Here, we have analyzed exhaled volatile metabolites (measured via real time mass spectrometry based breathomics) data from 204 healthy females, aged between 07 and 80 years. Age related substance-specific differences were observed in breath biomarkers. Exhalation of blood-borne endogenous organosulfur, short-chain fatty acids, alcohols, aldehydes, alkene, ketones and exogenous nitriles, terpenes, and aromatics have denominated interplay between endocrine differences, energy homeostasis, systemic microbial diversity, oxidative stress, and lifestyle. Overall marker expressions were suppressed under daily oral contraception. Young homosexual/lesbian adults turned out as breathomic outliers. Previously proposed disease-specific breath biomarkers should be reevaluated upon aging effects. Breathomics offers a noninvasive window toward system-wide understanding and personalized monitoring of aging i.e., translatable to gerontology.

Effects of COVID-19 protective face masks and wearing durations on respiratory haemodynamic physiology and exhaled breath constituents.

BACKGROUND: While assumed to protect against coronavirus transmission, face masks may have effects on respiratory-haemodynamic parameters. Within this pilot study, we investigated immediate and progressive effects of FFP2 and surgical masks on exhaled breath constituents and physiological attributes in 30 adults at rest. METHODS: We continuously monitored exhaled breath profiles within mask space in older (age 60-80 years) and young to middle-aged (age 20-59 years) adults over the period of 15 and 30 min by high-resolution real-time mass-spectrometry. Peripheral oxygen saturation (S pO2 ) and respiratory and haemodynamic parameters were measured (noninvasively) simultaneously. RESULTS: Profound, consistent and significant (p≤0.001) changes in S pO2 (≥60_FFP2-15 min: 5.8±1.3%↓, ≥60_surgical-15 min: 3.6±0.9%↓, <60_FFP2-30 min: 1.9±1.0%↓, <60_surgical-30 min: 0.9±0.6%↓) and end-tidal carbon dioxide tension (P ETCO2 ) (≥60_FFP2-15 min: 19.1±8.0%↑, ≥60_surgical-15 min: 11.6±7.6%↑, <60_FFP2- 30 min: 12.1±4.5%↑, <60_surgical- 30 min: 9.3±4.1%↑) indicate ascending deoxygenation and hypercarbia. Secondary changes (p≤0.005) to haemodynamic parameters (e.g. mean arterial pressure (MAP) ≥60_FFP2-15 min: 9.8±10.4%↑) were found. Exhalation of bloodborne volatile metabolites, e.g. aldehydes, hemiterpene, organosulfur, short-chain fatty acids, alcohols, ketone, aromatics, nitrile and monoterpene mirrored behaviour of cardiac output, MAP, S pO2 , respiratory rate and P ETCO2 . Exhaled humidity (e.g. ≥60_FFP2-15 min: 7.1±5.8%↑) and exhaled oxygen (e.g. ≥60_FFP2-15 min: 6.1±10.0%↓) changed significantly (p≤0.005) over time. CONCLUSIONS: Breathomics allows unique physiometabolic insights into immediate and transient effects of face mask wearing. Physiological parameters and breath profiles of endogenous and/or exogenous volatile metabolites indicated putative cross-talk between transient hypoxaemia, oxidative stress, hypercarbia, vasoconstriction, altered systemic microbial activity, energy homeostasis, compartmental storage and washout. FFP2 masks had a more pronounced effect than surgical masks. Older adults were more vulnerable to FFP2 mask-induced hypercarbia, arterial oxygen decline, blood pressure fluctuations and concomitant physiological and metabolic effects.

The peppermint breath test benchmark for PTR-MS and SIFT-MS.

A major challenge for breath research is the lack of standardization in sampling and analysis. To address this, a test that utilizes a standardized intervention and a defined study protocol has been proposed to explore disparities in breath research across different analytical platforms and to provide benchmark values for comparison. Specifically, thePeppermint Experimentinvolves the targeted analysis in exhaled breath of volatile constituents of peppermint oil after ingestion of the encapsulated oil. Data from thePeppermint Experimentperformed by proton transfer reaction mass spectrometry (PTR-MS) and selected ion flow tube mass spectrometry (SIFT-MS) are presented and discussed herein, including the product ions associated with the key peppermint volatiles, namely limonene,α- andß-pinene, 1,8-cineole, menthol, menthone and menthofuran. The breath washout profiles of these compounds from 65 individuals were collected, comprising datasets from five PTR-MS and two SIFT-MS instruments. The washout profiles of these volatiles were evaluated by comparing the log-fold change over time of the product ion intensities associated with each volatile. Benchmark values were calculated from the lower 95% confidence interval of the linear time-to-washout regression analysis for all datasets combined. Benchmark washout values from PTR-MS analysis were 353 min for the sum of monoterpenes and 1,8-cineole (identical product ions), 173 min for menthol, 330 min for menthofuran, and 218 min for menthone; from SIFT-MS analysis values were 228 min for the sum of monoterpenes, 281 min for the sum of monoterpenes and 1,8-cineole, and 370 min for menthone plus 1,8-cineole. Large inter- and intra-dataset variations were observed, whereby the latter suggests that biological variability plays a key role in how the compounds are absorbed, metabolized and excreted from the body via breath. This variability seems large compared to the influence of sampling and analytical procedures, but further investigations are recommended to clarify the effects of these factors.

Detection of Paratuberculosis in Dairy Herds by Analyzing the Scent of Feces, Alveolar Gas, and Stable Air.

Paratuberculosis is an important disease of ruminants caused by Mycobacterium avium ssp. paratuberculosis (MAP). Early detection is crucial for successful infection control, but available diagnostic tests are still dissatisfying. Methods allowing a rapid, economic, and reliable identification of animals or herds affected by MAP are urgently required. This explorative study evaluated the potential of volatile organic compounds (VOCs) to discriminate between cattle with and without MAP infections. Headspaces above fecal samples and alveolar fractions of exhaled breath of 77 cows from eight farms with defined MAP status were analyzed in addition to stable air samples. VOCs were identified by GC-MS and quantified against reference substances. To discriminate MAP-positive from MAP-negative samples, VOC feature selection and random forest classification were performed. Classification models, generated for each biological specimen, were evaluated using repeated cross-validation. The robustness of the results was tested by predicting samples of two different sampling days. For MAP classification, the different biological matrices emitted diagnostically relevant VOCs of a unique but partly overlapping pattern (fecal headspace: 19, alveolar gas: 11, stable air: 4-5). Chemically, relevant compounds belonged to hydrocarbons, ketones, alcohols, furans, and aldehydes. Comparing the different biological specimens, VOC analysis in fecal headspace proved to be most reproducible, discriminatory, and highly predictive.

Detection of Mycobacterium avium ssp. paratuberculosis in Cultures From Fecal and Tissue Samples Using VOC Analysis and Machine Learning Tools.

Analysis of volatile organic compounds (VOCs) is a novel approach to accelerate bacterial culture diagnostics of Mycobacterium avium subsp. paratuberculosis (MAP). In the present study, cultures of fecal and tissue samples from MAP-infected and non-suspect dairy cattle and goats were explored to elucidate the effects of sample matrix and of animal species on VOC emissions during bacterial cultivation and to identify early markers for bacterial growth. The samples were processed following standard laboratory procedures, culture tubes were incubated for different time periods. Headspace volume of the tubes was sampled by needle trap-micro-extraction, and analyzed by gas chromatography-mass spectrometry. Analysis of MAP-specific VOC emissions considered potential characteristic VOC patterns. To address variation of the patterns, a flexible and robust machine learning workflow was set up, based on random forest classifiers, and comprising three steps: variable selection, parameter optimization, and classification. Only a few substances originated either from a certain matrix or could be assigned to one animal species. These additional emissions were not considered informative by the variable selection procedure. Classification accuracy of MAP-positive and negative cultures of bovine feces was 0.98 and of caprine feces 0.88, respectively. Six compounds indicating MAP presence were selected in all four settings (cattle vs. goat, feces vs. tissue): 2-Methyl-1-propanol, 2-methyl-1-butanol, 3-methyl-1-butanol, heptanal, isoprene, and 2-heptanone. Classification accuracies for MAP growth-scores ranged from 0.82 for goat tissue to 0.89 for cattle feces. Misclassification occurred predominantly between related scores. Seventeen compounds indicating MAP growth were selected in all four settings, including the 6 compounds indicating MAP presence. The concentration levels of 2,3,5-trimethylfuran, 2-pentylfuran, 1-propanol, and 1-hexanol were indicative for MAP cultures before visible growth was apparent. Thus, very accurate classification of the VOC samples was achieved and the potential of VOC analysis to detect bacterial growth before colonies become visible was confirmed. These results indicate that diagnosis of paratuberculosis can be optimized by monitoring VOC emissions of bacterial cultures. Further validation studies are needed to increase the robustness of indicative VOC patterns for early MAP growth as a pre-requisite for the development of VOC-based diagnostic analysis systems.

Deficiency and absence of endogenous isoprene in adults, disqualified its putative origin.

BACKGROUND: Isoprene (C5H8) is a clinically important breath metabolite. Although, hundreds of studies have reported differential expressions in isoprene exhalation as breath biomarker for diverse diseases, the substance couldn't enter to clinical practice as diagnostic marker. Moreover, many experimental/basic observations upon breath isoprene remained unrelated to the corresponding pathophysiological effects on its putative metabolic origin (i.e. mevalonate pathway). Here, we investigated the fundamental reason that hindered the rational interpretation and translation of this marker from basic to clinical science. METHODS: Via high-resolution mass-spectrometry based breathomics in 1026 human subjects, we discovered adults with significant deficiency (order of magnitude lower than the normal) and complete absence of breath isoprene. We prospectively applied real-time breathomics, quantitative gene expression analysis of the mevalonate pathway enzymes, lipid-profiling and hemodynamic monitoring on those isoprene deficient subjects and controls. Additionally, the subject with absence of isoprene was followed up throughout different phases of her womanhood. RESULTS: In contrast to convention, we witnessed that adults can live healthy without exhaling isoprene or with significant deficiency. This rare phenotype represents a recessive inheritance. Despite physio-metabolic changes during menstrual cycle (that is known to profoundly affect isoprene exhalation) and profoundly increased plasma cholesterol during pregnancy and after childbirth, isoprene remained absent. All genes of mevalonate pathway enzymes were normally expressed in all participants, without any down-regulation or compensatory up-regulation. CONCLUSIONS: Absence/deficiency of isoprene despite normal lipid profiles and no mevalonate pathway malfunction disqualifies the long-believed metabolic origin of isoprene from cholesterol biosynthesis. Thus, clinical translation of breath isoprene expressions should not be generally attributed to corresponding pathophysiological effects onto mevalonate/cholesterol pathway. Our finding has refined and optimized the clinical interpretation of isoprene as biomarker in volatile metabolomics and breathomics. Future studies will address the correct metabolic origin of isoprene to imply this important marker to routine practice.

Spatial mapping of VOC exhalation by means of bronchoscopic sampling.

Breath analysis holds promise for non-invasive in vivo monitoring of disease related processes. However, physiological parameters may considerably affect profiles of exhaled volatile organic substances (VOCs). Volatile substances can be released via alveoli, bronchial mucosa or from the upper airways. The aim of this study was the systematic investigation of the influence of different sampling sites in the respiratory tract on VOC concentration profiles by means of a novel experimental setup. After ethical approval, breath samples were collected from 25 patients undergoing bronchoscopy for endobronchial ultrasound or bronchoscopic lung volume reduction from different sites in the airways. All patients had total intravenous anaesthesia under pressure-controlled ventilation. If necessary, respiratory parameters were adjusted to keep PETCO2 = 35-45 mm Hg. 30 ml gas were withdrawn at six sampling sites by means of gastight glass syringes: S1 = Room air, S2 = Inspiration, S3 = Endotracheal tube, S4 = Trachea, S5 = Right B6 segment, S6 = Left B6 segment (S4-S6 through the bronchoscope channel). 10 ml were used for VOC analysis, 20 ml for PCO2 determination. Samples were preconcentrated by solid-phase micro-extraction (SPME) and analysed by gas chromatography-mass spectrometry (GC-MS). PCO2 was determined in a conventional blood gas analyser. Statistically significant differences in substance concentrations for acetone, isoprene, 2-methyl-pentane and n-hexane could be observed between different sampling sites. Increasing substance concentrations were determined for acetone (15.3%), 2-methyl-pentane (11.4%) and n-hexane (19.3%) when passing from distal to proximal sampling sites. In contrast, isoprene concentrations decreased by 9.9% from proximal to more distal sampling sites. Blank bronchoscope measurements did not show any contaminations. Increased substance concentrations in the proximal respiratory tract may be explained through substance excretion from bronchial mucosa while decreased concentrations could result from absorption or reaction processes. Spatial mapping of VOC profiles can provide novel insights into substance specific exhalation kinetics and mechanisms.

Exhaled breath compositions under varying respiratory rhythms reflects ventilatory variations: translating breathomics towards respiratory medicine.

Control of breathing is automatic and its regulation is keen to autonomic functions. Therefore, involuntary and voluntary nervous regulation of breathing affects ventilatory variations, which has profound potential to address expanding challenges in contemporary pulmonology. Nonetheless, the fundamental attributes of the aforementioned phenomena are rarely understood and/or investigated. Implementation of unconventional approach like breathomics may leads to a better comprehension of those complexities in respiratory medicine. We applied breath-resolved spirometry and capnometry, non-invasive hemodynamic monitoring along with continuous trace analysis of exhaled VOCs (volatile organic compounds) by means of real-time mass-spectrometry in 25 young and healthy adult humans to investigate any possible mirroring of instant ventilatory variations by exhaled breath composition, under varying respiratory rhythms. Hemodynamics remained unaffected. Immediate changes in measured breath compositions and corresponding variations occurred when respiratory rhythms were switched between spontaneous (involuntary/unsynchronised) and/or paced (voluntary/synchronised) breathing. Such changes in most abundant, endogenous and bloodborne VOCs were closely related to the minute ventilation and end-tidal CO2 exhalation. Unprecedentedly, while preceded by a paced rhythm, spontaneous rhythms in both independent setups became reproducible with significantly (P-value ≤ 0.005) low intra- and inter-individual variation in measured parameters. We modelled breath-resolved ventilatory variations via alveolar isoprene exhalation, which were independently validated with unequivocal precision. Reproducibility i.e. attained via our method would be reliable for human breath sampling, concerning biomarker research. Thus, we may realize the actual metabolic and pathophysiological expressions beyond the everlasting in vivo physiological noise. Consequently, less pronounced changes are often misinterpreted as disease biomarker in cross-sectional studies. We have also provided novel information beyond conventional spirometry and capnometry. Upon clinical translations, our findings will have immense impact on pulmonology and breathomics as they have revealed a reproducible pattern of ventilatory variations and respiratory homeostasis in endogenous VOC exhalations.

Effects of modular ion-funnel technology onto analysis of breath VOCs by means of real-time mass spectrometry.

Proton transfer reaction time-of-flight mass spectrometry (PTR-ToF-MS) is a powerful tool for real-time monitoring of trace concentrations of volatile organic compounds (VOCs). The sensitivity of PTR-ToF-MS also depends on the ability to effectively focus and transmit ions from the relatively high-pressure drift tube (DT) to the low-pressure mass analyzer. In the present study, a modular ion-funnel (IF) is placed adjacent to the DT of a PTR-ToF-MS instrument to improve the ion-focusing. IF consists of a series of electrodes with gradually decreasing orifice diameters. Radio frequency (RF) voltage and direct current (DC) electric field are then applied to the electrodes to get the ions focused. We investigated the effect of the RF voltage and DC field on the sensitivity of a pattern of VOCs including hydrocarbons, alcohols, aldehydes, ketones, and aromatic compounds. In a proof-of-concept study, the instrument operating both as normal DT (DC-mode) and at optimal IF conditions (RF-mode) was applied for the breath analysis of 21 healthy human subjects. For the range of investigated VOCs, an improvement of one order of magnitude in sensitivity was observed in RF-mode compared with DC-mode. Limits of detection could be improved by a factor of 2-4 in RF-mode compared with DC-mode. Operating the instrument in RF-mode allowed the detection of more compounds in the exhaled air compared with DC-mode. Incorporation of the IF considerably improved the performance of PTR-ToF-MS allowing the real-time monitoring of a larger number of potential breath biomarkers. Graphical abstract.

Changes of Exhaled Volatile Organic Compounds in Postoperative Patients Undergoing Analgesic Treatment: A Prospective Observational Study.

Assessment and treatment of postoperative pain can be challenging as objective examination techniques to detect and quantify pain are lacking. We aimed to investigate changes of exhaled volatile organic compounds (VOCs) in patients with postoperative pain before and after treatment with opioid analgesics. In an observational study in 20 postoperative patients, we monitored for postoperative pain, hemodynamic parameters, and catecholamines before and during treatment. VOCs in the patients were determined by direct real-time proton transfer reaction time-of-flight mass spectrometry prior (0 min) and after piritramide application (15 min as well as 30 min). Cardiovascular variables changed and norepinephrine levels decreased during treatment. The VOCs acetonitrile (<0.001), acetaldehyde (p = 0.002), benzopyran (p = 0.004), benzene (p < 0.001), hexenal (p = < 0.001), 1-butanethiol (p = 0.004), methanethiol (p < 0.001), ethanol (p = 0.003), and propanol (p = < 0.001) changed significantly over time. Patients with Numeric Rating Scale (NRS) < 4 showed a significantly lower concentration of hexenal compared to patients with NRS > 4 at the time points 15 min (45.0 vs. 385.3 ncps, p = 0.047) and 30 min (38.3 vs. 334.6 ncps, p = 0.039). Breath analysis can provide additional information for noninvasive monitoring for analgesic treatment in postoperative patients.

Author Correction: Exhaled volatile substances in children suffering from type 1 diabetes mellitus: results from a cross-sectional study.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.