Modelling of Breath and Various Blood Volatilomic Profiles-Implications for Breath Volatile Analysis.

Researchers looking for biomarkers from different sources, such as breath, urine, or blood, frequently search for specific patterns of volatile organic compounds (VOCs), often using pattern recognition or machine learning techniques. However, they are not generally aware that these patterns change depending on the source they use. Therefore, we have created a simple model to demonstrate that the distribution patterns of VOCs in fat, mixed venous blood, alveolar air, and end-tidal breath are different. Our approach follows well-established models for the description of dynamic real-time breath concentration profiles. We start with a uniform distribution of end-tidal concentrations of selected VOCs and calculate the corresponding target concentrations. For this, we only need partition coefficients, mass balance, and the assumption of an equilibrium state, which avoids the need to know the volatiles' metabolic rates and production rates within the different compartments.

Blood and breath profiles of volatile organic compounds in patients with end-stage renal disease.

BACKGROUND: Monitoring of volatile organic compounds (VOCs) in exhaled breath shows great potential as a non-invasive method for assessing hemodialysis efficiency. In this work we aim at identifying and quantifying of a wide range of VOCs characterizing uremic breath and blood, with a particular focus on species responding to the dialysis treatment. METHODS: Gas chromatography with mass spectrometric detection coupled with solid-phase microextraction as pre-concentration method. RESULTS: A total of 60 VOCs were reliably identified and quantified in blood and breath of CKD patients. Excluding contaminants, six compounds (isoprene, dimethyl sulfide, methyl propyl sulfide, allyl methyl sulfide, thiophene and benzene) changed their blood and breath levels during the hemodialysis treatment. CONCLUSIONS: Uremic breath and blood patterns were found to be notably affected by the contaminants from the extracorporeal circuits and hospital room air. Consequently, patient exposure to a wide spectrum of volatile species (hydrocarbons, aldehydes, ketones, aromatics, heterocyclic compounds) is expected during hemodialysis. Whereas highly volatile pollutants were relatively quickly removed from blood by exhalation, more soluble ones were retained and contributed to the uremic syndrome. At least two of the species observed (cyclohexanone and 2-propenal) are uremic toxins. Perhaps other volatile substances reported within this study may be toxic and have negative impact on human body functions. Further studies are required to investigate if VOCs responding to HD treatment could be used as markers for monitoring hemodialysis efficiency.

Release and uptake of volatile organic compounds by human hepatocellular carcinoma cells (HepG2) in vitro.

BACKGROUND: Volatile organic compounds (VOCs) emitted by human body offer a unique insight into biochemical processes ongoing in healthy and diseased human organisms. Unfortunately, in many cases their origin and metabolic fate have not been yet elucidated in sufficient depth, thus limiting their clinical application. The primary goal of this work was to identify and quantify volatile organic compounds being released or metabolized by HepG2 hepatocellular carcinoma cells. METHODS: The hepatocellular carcinoma cells were incubated in specially designed head-space 1-L glass bottles sealed for 24 hours prior to measurements. Identification and quantification of volatiles released and consumed by cells under study were performed by gas chromatography with mass spectrometric detection (GC-MS) coupled with head-space needle trap device extraction (HS-NTD) as the pre-concentration technique. Most of the compounds were identified both by spectral library match as well as retention time comparison based on standards. RESULTS: A total of nine compounds were found to be metabolised and further twelve released by the cells under study (Wilcoxon signed-rank test, p<0.05). The former group comprised 6 aldehydes (2-methyl 2-propenal, 2-methyl propanal, 2-ethylacrolein, 3-methyl butanal, n-hexanal and benzaldehyde), n-propyl propionate, n-butyl acetate, and isoprene. Amongst the released species there were five ketones (2-pentanone, 3-heptanone, 2-heptanone, 3-octanone, 2-nonanone), five volatile sulphur compounds (dimethyl sulfide, ethyl methyl sulfide, 3-methyl thiophene, 2-methyl-1-(methylthio)- propane and 2-methyl-5-(methylthio) furan), n-propyl acetate, and 2-heptene. CONCLUSIONS: The emission and uptake of the aforementioned VOCs may reflect the activity of abundant liver enzymes and support the potential of VOC analysis for the assessment of enzymes function.

Stability of selected volatile breath constituents in Tedlar, Kynar and Flexfilm sampling bags.

The stability of 41 selected breath constituents in three types of polymer sampling bags, Tedlar, Kynar, and Flexfilm, was investigated using solid phase microextraction and gas chromatography mass spectrometry. The tested molecular species belong to different chemical classes (hydrocarbons, ketones, aldehydes, aromatics, sulphurs, esters, terpenes, etc.) and exhibit close-to-breath low ppb levels (3-12 ppb) with the exception of isoprene, acetone and acetonitrile (106 ppb, 760 ppb, 42 ppb respectively). Stability tests comprised the background emission of contaminants, recovery from dry samples, recovery from humid samples (RH 80% at 37 °C), influence of the bag's filling degree, and reusability. Findings yield evidence of the superiority of Tedlar bags over remaining polymers in terms of background emission, species stability (up to 7 days for dry samples), and reusability. Recoveries of species under study suffered from the presence of high amounts of water (losses up to 10%). However, only heavier volatiles, with molecular masses higher than 90, exhibited more pronounced losses (20-40%). The sample size (the degree of bag filling) was found to be one of the most important factors affecting the sample integrity. To sum up, it is recommended to store breath samples in pre-conditioned Tedlar bags up to 6 hours at the maximum possible filling volume. Among the remaining films, Kynar can be considered as an alternative to Tedlar; however, higher losses of compounds should be expected even within the first hours of storage. Due to the high background emission Flexfilm is not suitable for sampling and storage of samples for analyses aiming at volatiles at a low ppb level.

Blood and breath levels of selected volatile organic compounds in healthy volunteers.

Gas chromatography with mass spectrometric detection (GC-MS) was used to identify and quantify volatile organic compounds in the blood and breath of healthy individuals. Blood and breath volatiles were pre-concentrated using headspace solid phase micro-extraction (HS-SPME) and needle trap devices (NTDs), respectively. The study involved a group of 28 healthy test subjects and resulted in the quantification of a total of 74 compounds in both types of samples. The concentrations of the species under study varied between 0.01 and 6700 nmol L(-1) in blood and between 0.02 and 2500 ppb in exhaled air. Limits of detection (LOD) ranged from 0.01 to 270 nmol L(-1) for blood compounds and from 0.01 to 0.7 ppb for breath species. Relative standard deviations for both measurement regimes varied from 1.5 to 14%. The predominant chemical classes among the compounds quantified were hydrocarbons (24), ketones (10), terpenes (8), heterocyclic compounds (7) and aromatic compounds (7). Twelve analytes were found to be highly present in both blood and exhaled air (with incidence rates higher than 80%) and for 32 species significant differences (Wilcoxon signed-rank test) between room air and exhaled breath were observed. By comparing blood, room air and breath levels in parallel, a tentative classification of volatiles into endogenous and exogenous compounds can be achieved.

Human blood and plasma partition coefficients for C4-C8 n-alkanes, isoalkanes, and 1-alkenes.

Human blood:air and plasma:air partition coefficients for C(4)-C(8) n-alkanes, isoalkanes, and 1-alkenes were determined using multiple headspace extraction coupled with solid phase microextraction and gas chromatography. Mean blood:air partition coefficients expressed in the form of dimensionless blood-to-air concentration ratio (g/mL(b)/g/mL(a)) were 0.183, 0.416, 1.08, 2.71, and 5.77 for C(4)-C(8) n-alkanes; 0.079, 0.184, 0.473, 1.3, and 3.18 for C(4)-C(8) isoalkanes; and 0.304, 0.589, 1.32, 3.5, and 7.01 for C(4)-C(8) 1-alkenes, respectively (n = 8). The reported partition coefficient values increased exponentially with boiling points, molecular weights, and the carbon atoms in the particle. The solubility of 1-alkenes in blood was higher than in plasma, whereas the blood:air and plasma:air partition coefficients of n-alkanes and isoalkanes did not differ significantly. Consequently, additional interactions of 1-alkenes with whole blood seem to occur. The presented findings are expected to be particularly useful for assessing the uptake, distribution, and elimination of hydrocarbons in human organism.

A mathematical model for breath gas analysis of volatile organic compounds with special emphasis on acetone.

Recommended standardized procedures for determining exhaled lower respiratory nitric oxide and nasal nitric oxide (NO) have been developed by task forces of the European Respiratory Society and the American Thoracic Society. These recommendations have paved the way for the measurement of nitric oxide to become a diagnostic tool for specific clinical applications. It would be desirable to develop similar guidelines for the sampling of other trace gases in exhaled breath, especially volatile organic compounds (VOCs) which may reflect ongoing metabolism. The concentrations of water-soluble, blood-borne substances in exhaled breath are influenced by: (i) breathing patterns affecting gas exchange in the conducting airways, (ii) the concentrations in the tracheo-bronchial lining fluid, (iii) the alveolar and systemic concentrations of the compound. The classical Farhi equation takes only the alveolar concentrations into account. Real-time measurements of acetone in end-tidal breath under an ergometer challenge show characteristics which cannot be explained within the Farhi setting. Here we develop a compartment model that reliably captures these profiles and is capable of relating breath to the systemic concentrations of acetone. By comparison with experimental data it is inferred that the major part of variability in breath acetone concentrations (e.g., in response to moderate exercise or altered breathing patterns) can be attributed to airway gas exchange, with minimal changes of the underlying blood and tissue concentrations. Moreover, the model illuminates the discrepancies between observed and theoretically predicted blood-breath ratios of acetone during resting conditions, i.e., in steady state. Particularly, the current formulation includes the classical Farhi and the Scheid series inhomogeneity model as special limiting cases and thus is expected to have general relevance for a wider range of blood-borne inert gases. The chief intention of the present modeling study is to provide mechanistic relationships for further investigating the exhalation kinetics of acetone and other water-soluble species. This quantitative approach is a first step towards new guidelines for breath gas analyses of volatile organic compounds, similar to those for nitric oxide.

Expanding theory-based evaluation: Incorporating value creation in a theory of change.

Evaluations of policies and programs often use a theory of change to articulate how the intervention is intended to function and the mechanisms by which it is supposed to generate outcomes. When an evaluation includes cost and efficiency considerations, economic and other concepts can be added to a theory of change to articulate a theory of value creation that articulates the mechanisms by which the intervention should use resources efficiently, effectively and create sufficient value to justify the resource use. This paper introduces some theories of value creation that are often implicit in program designs. Making these theories explicit can support clearer evaluative thinking about value for money - including specification of criteria and standards that are aligned with the theory, methods of inquiry that test the theory, and well-reasoned judgements that answer evaluative questions about value for money. Implications for evaluation practice will be discussed.

Physiological modeling of isoprene dynamics in exhaled breath.

Human breath contains a myriad of endogenous volatile organic compounds (VOCs) which are reflective of ongoing metabolic or physiological processes. While research into the diagnostic potential and general medical relevance of these trace gases is conducted on a considerable scale, little focus has been given so far to a sound analysis of the quantitative relationships between breath levels and the underlying systemic concentrations. This paper is devoted to a thorough modeling study of the end-tidal breath dynamics associated with isoprene, which serves as a paradigmatic example for the class of low-soluble, blood-borne VOCs. Real-time measurements of exhaled breath under an ergometer challenge reveal characteristic changes of isoprene output in response to variations in ventilation and perfusion. Here, a valid compartmental description of these profiles is developed. By comparison with experimental data it is inferred that the major part of breath isoprene variability during exercise conditions can be attributed to an increased fractional perfusion of potential storage and production sites, leading to higher levels of mixed venous blood concentrations at the onset of physical activity. In this context, various lines of supportive evidence for an extrahepatic tissue source of isoprene are presented. Our model is a first step towards new guidelines for the breath gas analysis of isoprene and is expected to aid further investigations regarding the exhalation, storage, transport and biotransformation processes associated with this important compound.

Effect of inhaled acetone concentrations on exhaled breath acetone concentrations at rest and during exercise.

Real-time measurements of the differences in inhaled and exhaled, unlabeled and fully deuterated acetone concentration levels, at rest and during exercise, have been conducted using proton transfer reaction mass spectrometry. A novel approach to continuously differentiate between the inhaled and exhaled breath acetone concentration signals is used. This leads to unprecedented fine grained data of inhaled and exhaled concentrations. The experimental results obtained are compared with those predicted using a simple three compartment model that theoretically describes the influence of inhaled concentrations on exhaled breath concentrations for volatile organic compounds with high blood:air partition coefficients, and hence is appropriate for acetone. An agreement between the predicted and observed concentrations is obtained. Our results highlight that the influence of the upper airways cannot be neglected for volatiles with high blood:air partition coefficients, i.e. highly water soluble volatiles.

Toward harmonizing prospective effectiveness assessment for road safety: Comparing tools in standard test case simulations.

Objective: With the overall goal to harmonize prospective effectiveness assessment of active safety systems, the specific objective of this study is to identify and evaluate sources of variation in virtual precrash simulations and to suggest topics for harmonization resulting in increased comparability and thus trustworthiness of virtual simulation-based prospective effectiveness assessment. Methods: A round-robin assessment of the effectiveness of advanced driver assistance systems was performed using an array of state-of-the-art virtual simulation tools on a set of standard test cases. The results were analyzed to examine reasons for deviations in order to identify and assess aspects that need to be harmonized and standardized. Deviations between results calculated by independent engineering teams using their own tools should be minimized if the research question is precisely formulated regarding input data, models, and postprocessing steps. Results: Two groups of sources of variations were identified; one group (mostly related to the implementation of the system under test) can be eliminated by using a more accurately formulated research question, whereas the other group highlights further harmonization needs because it addresses specific differences in simulation tool setups. Time-to-collision calculations, vehicle dynamics, especially braking behavior, and hit-point position specification were found to be the main sources of variation. Conclusions: The study identified variations that can arise from the use of different simulation setups in assessment of the effectiveness of active safety systems. The research presented is a first of its kind and provides significant input to the overall goal of harmonization by identifying specific items for standardization. Future activities aim at further specification of methods for prospective assessments of the effectiveness of active safety, which will enhance comparability and trustworthiness in this kind of studies and thus contribute to increased traffic safety.

Modeling-based determination of physiological parameters of systemic VOCs by breath gas analysis, part 2.

In a recent paper (Unterkofler et al 2015 J. Breath Res. 9 036002) we presented a simple two compartment model which describes the influence of inhaled concentrations on exhaled breath concentrations for volatile organic compounds (VOCs) with small Henry constants. In this paper we extend this investigation concerning the influence of inhaled concentrations on exhaled breath concentrations for VOCs with higher Henry constants. To this end we extend our model with an additional compartment which takes into account the influence of the upper airways on exhaled breath VOC concentrations.

Nurse-led school-based clinics for rheumatic fever prevention and skin infection management: evaluation of Mana Kidz programme in Counties Manukau.

AIM: To evaluate registered nurse-led school clinics in 61 primary and intermediate schools in Counties Manukau. METHODS: The evaluation (conducted August­December, 2014) collated evidence concerning service delivery, outcomes, value for money and effectiveness. RESULTS: 97% (23,756/24,497) of eligible children were consented, 11% (20,696/191,423) of throat swabs taken (February 2013­September 2014) were culture positive for Group A Streptococcus (GAS); 20,176 were treated. Mana Kidz teams treated (includes cleaning and covering alone) 17,593 skin infections and actioned 4,178 school health referrals. A pre-programme cross sectional GAS pharyngeal prevalence demonstrated a relative risk 1.8 (1.3­2.3) (95%CI) of being pharyngeal GAS positive in 2013 compared to 2014. Hospitalisations for acute rheumatic fever (ARF) and skin infections for children aged 5­12 years living in Counties Manukau are declining and this appears to be temporally related to the introduction of the Mana Kidz programme. Effective engagement with children, parents/ whanau and improved health literacy was demonstrated, especially knowledge about sore throats, ARF, medication adherence and skin infection. The programme was delivered at $280 per participating child in the 2013/14 financial year. CONCLUSION: Mana Kidz is an effective programme with a substantial contribution to health care for children, aged 5­12 years, identified at increased risk of poor health outcomes.

Modeling of breath methane concentration profiles during exercise on an ergometer.

We develop a simple three compartment model based on mass balance equations which quantitatively describes the dynamics of breath methane concentration profiles during exercise on an ergometer. With the help of this model it is possible to estimate the endogenous production rate of methane in the large intestine by measuring breath gas concentrations of methane.

The effect of alcohol price on dependent drinkers' alcohol consumption.

AIMS: To investigate the current purchasing behaviours of a group of dependent drinkers and their potential response to future increases in the price of alcohol. METHODS: 115 clients undergoing medical detoxification completed an anonymous survey about their daily alcohol consumption, its cost, their response to potential price increases and strategies previously used when unable to afford alcohol. RESULTS: Mean and median number of standard drinks consumed per day was 24, at a median cost of $25 NZD (95%CI $22, $30). Thirty-six per cent (95%CI 26%, 46%) of the group bought alcohol at $1 or less per standard drink, and the median number of drinks consumed per day (30) by this group was significantly higher (p=0.0028) than the rest of the sample (22.5). The most common strategy used if no money was available to purchase alcohol was to forgo essentials. If facing a potential price rise, 77% (95%CI 69%, 85%) would switch wholly or partially to a cheaper product and 13% (95%CI 8%, 21%) would cut down their drinking. CONCLUSIONS: Although the majority of our group would be financially impacted by an increase in the minimum price per standard drink, any potential impacts would be most significant in those buying the cheapest alcohol (who also drink the most), suggesting that minimum pricing may be an important harm minimisation strategy in this group. A minimum price per standard drink would limit the possibility of switching to an alternate cheaper product and likely result in an overall reduction in alcohol consumption in this group. Stealing alcohol, or the use of non-beverage alcohol, were seldom reported as previous strategies used in response to unaffordable alcohol and fears of such are not valid reasons for rejecting minimum pricing to reduce general population consumption.

Modeling-based determination of physiological parameters of systemic VOCs by breath gas analysis: a pilot study.

In this paper we develop a simple two compartment model which extends the Farhi equation to the case when the inhaled concentration of a volatile organic compound (VOC) is not zero. The model connects the exhaled breath concentration of systemic VOCs with physiological parameters such as endogenous production rates and metabolic rates. Its validity is tested with data obtained for isoprene and inhaled deuterated isoprene-D5.

Emission rates of selected volatile organic compounds from skin of healthy volunteers.

Gas chromatography with mass spectrometric detection (GC-MS) coupled with solid phase micro-extraction as pre-concentration method (SPME) was applied to identify and quantify volatile organic compounds (VOCs) emitted by human skin. A total of 64 C4-C10 compounds were quantified in skin emanation of 31 healthy volunteers. Amongst them aldehydes and hydrocarbons were the predominant chemical families with eighteen and seventeen species, respectively. Apart from these, there were eight ketones, six heterocyclic compounds, six terpenes, four esters, two alcohols, two volatile sulphur compounds, and one nitrile. The observed median emission rates ranged from 0.55 to 4,790 fmol cm(-2)min(-1). Within this set of analytes three volatiles; acetone, 6-methyl-5-hepten-2-one, and acetaldehyde exhibited especially high emission rates exceeding 100 fmol cm(-2)min(-1). Thirty-three volatiles were highly present in skin emanation with incidence rates over 80%. These species can be considered as potential markers of human presence, which could be used for early location of entrapped victims during Urban Search and Rescue Operations (USaR).

ABA-Cloud: support for collaborative breath research.

This paper introduces the advanced breath analysis (ABA) platform, an innovative scientific research platform for the entire breath research domain. Within the ABA project, we are investigating novel data management concepts and semantic web technologies to document breath analysis studies for the long run as well as to enable their full automatic reproducibility. We propose several concept taxonomies (a hierarchical order of terms from a glossary of terms), which can be seen as a first step toward the definition of conceptualized terms commonly used by the international community of breath researchers. They build the basis for the development of an ontology (a concept from computer science used for communication between machines and/or humans and representation and reuse of knowledge) dedicated to breath research.

Measurement of endogenous acetone and isoprene in exhaled breath during sleep.

This explorative study aims at characterizing the breath behavior of two prototypic volatile organic compounds, acetone and isoprene, during normal human sleep and to possibly relate changes in the respective concentration time courses to the underlying sleep architecture. For this purpose, six normal healthy volunteers (two females, four males, age 20-29 years) were monitored over two consecutive nights (the first one being an adaption night) by combining real-time proton-transfer-reaction mass spectrometry measurements from end-tidal exhalation segments with laboratory-based polysomnographic data. Breath acetone concentrations increased overnight in all measurements, with an average relative change by a factor of up to 4 (median 2.5). Nighttime concentration maxima were usually recorded 2-3 h before lights on. For breath isoprene, a nocturnal increase in baseline concentrations of about 74% was observed, with individual changes ranging from 36-110%. Isoprene profiles exhibited pronounced concentration peaks, which were highly specific for leg movements as scored by tibial electromyography. Furthermore, relative to a linear trend, baseline isoprene concentrations decreased during the transition from the NREM to the REM phase of a complete sleep cycle.

Breath gas analysis for estimating physiological processes using anesthetic monitoring as a prototypic example.

Analysis of exhaled trace gases is a novel methodology for gaining continuous and non-invasive information on the clinical state of an individual. This paper serves to explore some potential applications of breath gas analysis in anesthesia, describing a monitoring scheme for target site concentrations and cardiac output via physiological modeling and real-time breath profiles of the anesthetic agent. The rationale given here is mainly simulation-based, however, the underlying concepts are directly applicable to a routine clinical setting.