Metabolomics studies are becoming increasingly common for understanding how plant metabolism responds to changes in environmental conditions, genetic manipulations and treatments. Despite the recent advances in metabolomics workflow, the sample preparation process still limits the high-throughput analysis in large-scale studies. Here, we present a highly flexible robotic system that integrates liquid handling, sonication, centrifugation, solvent evaporation and sample transfer processed in 96-well plates to automatize the metabolite extraction from leaf samples. We transferred an established manual extraction protocol performed to a robotic system, and with this, we show the optimization steps required to improve reproducibility and obtain comparable results in terms of extraction efficiency and accuracy. We then tested the robotic system to analyze the metabolomes of wild-type and four transgenic silver birch (Betula pendula Roth) lines under unstressed conditions. Birch trees were engineered to overexpress the poplar (Populus × canescens) isoprene synthase and to emit various amounts of isoprene. By fitting the different isoprene emission capacities of the transgenic trees with their leaf metabolomes, we observed an isoprene-dependent upregulation of some flavonoids and other secondary metabolites as well as carbohydrates, amino acid and lipid metabolites. By contrast, the disaccharide sucrose was found to be strongly negatively correlated to isoprene emission. The presented study illustrates the power of integrating robotics to increase the sample throughput, reduce human errors and labor time, and to ensure a fully controlled, monitored and standardized sample preparation procedure. Due to its modular and flexible structure, the robotic system can be easily adapted to other extraction protocols for the analysis of various tissues or plant species to achieve high-throughput metabolomics in plant research.

Isoprene is one of the most abundant and most frequently evaluated volatile organic compounds in exhaled breath. Recently, several individuals with background levels of exhaled isoprene have been identified. Here, case study data are provided for an individual, identified from a previous study, with this low prevalence phenotype. It is hypothesized that the individual will illustrate low levels of exhaled isoprene at rest and during exercise. At rest, the subject (7.1 ppb) shows background (μ= 14.2 ± 7.0 ppb) levels of exhaled isoprene while the control group illustrates significantly higher quantities (μ= 266.2 ± 72.3 ppb) via proton transfer reaction mass spectrometry (PTR-MS). The result, background levels of isoprene at rest, is verified by thermal desorption gas chromatography mass spectrometry (TD-GC-MS) collections with the individual showing -3.6 ppb exhaled isoprene while the room background containedμ= -4.1 ± 0.1 ppb isoprene. As isoprene has been shown previously to increase at the initiation of exercise, exercise bike experiments were performed with the individual identified with low isoprene, yielding low and invariant levels of exhaled isoprene (μ= 6.6 ± 0.1 ppb) during the exercise while control subjects illustrated an approximate 2.5-fold increase (preμ= 286.3 ± 43.8 ppb, exerciseμ= 573.0 ± 147.8 ppb) in exhaled isoprene upon exercise start. Additionally, exhaled breath bag data showed a significant decrease in isoprene (delta post/pre, p = 0.0078) of the control group following the exercise regimen. Finally, TD-GC-MS results for exhaled isoprene from the individual\'s family (mother, father, sister and maternal grandmother) illustrated that the mother and father exhibited isoprene values (28.5 ppb, 77.2 ppb) below control samples 95% confidence interval (μ= 166.8 ± 43.3 ppb) while the individual\'s sister (182.0 ppb) was within the control range. These data provide evidence for a large dynamic range in exhaled isoprene in this family. Collectively, these results provide additional data surrounding the existence of a small population of individuals with background levels of exhaled isoprene.

Theoretical consideration about the impact of methyl groups on the structure and vibrational properties of β-carotenoids, using medium size molecules of trans-butadiene and trans-isoprene, are reported. Density functional theory (DFT) calculations with correlation-consistent and polarization-consistent basis sets were applied to trans-1,3-butadiene and trans-isoprene as the smallest building bricks of β-carotenoids. Their structure and harmonic vibrations were estimated in the complete basis set limit (CBS) using the non-linear least square fit. Optimized geometries and harmonic frequencies, obtained with B3LYP and BLYP density functionals and large basis sets, were favorably reproduced by a significantly faster approach, using a recently modified STO(1M)-3G Slater-type basis set. Selected density functionals with STO(1M)-3G and 6-311++G** basis sets were also successful in predicting β-carotene structures and harmonic vibrations. This work demonstrates the potential applicability of the proposed level of theory for larger molecules, including β-carotenoids, present in numerous natural food sources. The proposed scheme of molecular modeling, applied to biologically active compounds in food, could provide a deeper insight into their function in vivo, which is directly related to their structure and spectroscopic properties. It could also support the experimental qualitative analysis, based on peak assignment of β-carotenoids in various food sources.

Particle Image Velocimetry (PIV) measurements with the aim of providing experimental data for the SimInhale benchmark case are presented within this work. We, therefore, present a refractive index matched, transparent model of the benchmark geometry, in which the velocity and turbulent kinetic energy fields are examined at flow rates comparable to 15, 30 and 60 L/min (Re ≈ 1000-4500) in air. Furthermore, these results are compared with Large Eddy Simulations (LES). The results reveal a Reynolds number independence of the qualitative velocity field in the range covered within this work. Good agreement is found between the PIV and LES data, with a slight over-prediction of turbulent kinetic energies by the simulations. The obtained experimental data will be part of a common, publicly accessible ERCOFTAC database along with additional results published recently.

Remediation of dense non-aqueous phase liquids (DNAPLs) represents a challenging issue because of their persistent behaviour in the environment. This pilot-scale study investigates, by means of in situ experiments and numerical modelling, the feasibility of the pulsed pumping process of a large amount of a DNAPL in an alluvial aquifer. The main compound of the DNAPL is hexachlorobutadiene (HCBD), added in 2015 to the persistent organic pollutants list (POP). A low-permeability keyed enclosure was built at the location of the DNAPL source zone in order to isolate a finite volume of soil and a 3-month pulsed pumping process was applied inside the enclosure to exclusively extract the DNAPL. The water/DNAPL interface elevation at both the pumping well and an observation well was recorded. The cumulated pumped volume of DNAPL was also monitored. A total volume of about 20 m3 of pure DNAPL was recovered since no water was extracted during the process. The three-dimensional and multiphase flow simulator TMVOC was used and a conceptual model was elaborated and generated with the pre/post-processing tool mView. Numerical simulations reproduce the pulsed pumping process and show an excellent match between simulated and field data of DNAPL cumulated pumped volume and a reasonable agreement between modelled and observed data for the evolution of the water/DNAPL interface elevations at the two wells. This study offers a new perspective in remediation since DNAPL pumping system optimisation may be performed where a large amount of DNAPL is encountered.

The potential role of isoprene oxidative processes, as well as the possible impact of air pollution on isoprene emissions, are more important in tropical cities, surrounded by rainforests. In this study, the contribution of isoprene to ozone formation was determined considering different scenarios, mainly volatile organic compounds/NO x (VOC/NO x ) ratios, and typical atmospheric conditions for the city of Rio de Janeiro, where more than 36% of the urbanized area is covered by vegetation. Ozone isopleths and incremental reactivity coefficients (IR) were evaluated to understand the direct contribution of isoprene to ground-level ozone formation and the negative impact of anthropogenic NO x emissions on the natural atmospheric balance. Although isoprene accounted for only 2.7% of the total VOC mass, excluding the isoprene concentration from the model reduced the maximum ozone value by 14.1%. The calculated IR coefficient (grams of O3 formed per gram of added isoprene) was 2.2 for a VOC/NO x ratio of 8.86.

In the efforts at controlling automobile emissions, it is important to know in what extent air pollutants from on-road vehicles could be truly reduced. In 2014 we conducted tests in a heavily trafficked tunnel in south China to characterize emissions of volatile organic compounds (VOC) from on-road vehicle fleet and compared our results with those obtained in the same tunnel in 2004. Alkanes, aromatics, and alkenes had average emission factors (EFs) of 338, 63, and 42 mg km-1 in 2014 against that of 194, 129, and 160 mg km-1 in 2004, respectively. In 2014, LPG-related propane, n-butane and i-butane were the top three non-methane hydrocarbons (NMHCs) with EFs of 184 ± 21, 53 ± 6 and 31 ± 3 mg km-1; the gasoline evaporation marker i-pentane had an average EF of 17 ± 3 mg km-1; ethylene and propene were the top two alkenes with average EFs of 16 ± 1 and 9.7 ± 0.9 mg km-1, respectively; isoprene had no direct emission from vehicles; toluene showed the highest EF of 11 ± 2 mg km-1 among the aromatics; and acetylene had an average EF of 7 ± 1 mg km-1. While EFs of total NMHCs decreased only 9% from 493 ± 120 mg km-1 in 2004 to 449 ± 40 mg km-1 in 2014, their total ozone formation potential (OFP) decreased by 57% from 2.50 × 103 mg km-1 in 2004 to 1.10 × 103 mg km-1 in 2014, and their total secondary organic aerosol formation potential (SOAFP) decreased by 50% from 50 mg km-1 in 2004 to 25 mg km-1 in 2014. The large drop in ozone and SOA formation potentials could be explained by reduced emissions of reactive alkenes and aromatics, due largely to fuel transition from gasoline/diesel to LPG for taxis/buses and upgraded vehicle emission standards.

UNASSIGNED: Three-dimensional (3D) printing is being increasingly used in manufacturing and by small business entrepreneurs and home hobbyists. Exposure to airborne emissions during 3D printing raises the issue of whether there may be adverse health effects associated with these emissions.
UNASSIGNED: We present a case of a worker who developed asthma while using 3D printers, which illustrates that respiratory problems may be associated with 3D printer emissions.
UNASSIGNED: The patient was a 28-year-old self-employed businessman with a past history of asthma in childhood, which had resolved completely by the age of eight. He started using 10 fused deposition modelling 3D printers with acrylonitrile-butadiene-styrene filaments in a small work area of approximately 3000 cubic feet. Ten days later, he began to experience recurrent chest tightness, shortness of breath and coughing at work. After 3 months, his work environment was modified by reducing the number of printers, changing to polylactic acid filaments and using an air purifier with an high-efficiency particulate air filter and organic cartridge. His symptoms improved gradually, although he still needed periodic treatment with a salbutamol inhaler. While still symptomatic, a methacholine challenge indicated a provocation concentration causing a 20% fall in FEV1 (PC20) of 4 mg/ml, consistent with mild asthma. Eventually, his symptoms resolved completely and a second methacholine challenge after symptom resolution was normal (PC20 > 16 mg/ml).
UNASSIGNED: This case indicates that workers may develop respiratory problems, including asthma when using 3D printers. Further investigation of the specific airborne emissions and health problems from 3D printing is warranted.

Traffic exhaust, refineries and industrial facilities are major sources of air toxics identified by the U.S. Environmental Protection Agency (U.S. EPA) for their potential risk to human health. In utero and early life exposures to air toxics such as benzene and 1,3-butadiene, which are known leukemogens in adults, may play an etiologic role in childhood leukemia that comprises the majority of pediatric cancers. We conducted a population based case-control study to examine individual effects of benzene, 1,3-butadiene and polycyclic organic matter (POM) in ambient residential air on acute lymphocytic leukemia (ALL) diagnosed in children under age 5 years in Texas from 1995-2011.
Texas Cancer Registry cases were linked to birth records and then were frequency matched by birth month and year to 10 population-based controls. Maternal and infant characteristics from birth certificates were abstracted to obtain information about potential confounders. Modelled estimates of benzene, 1,3-butadiene and POM exposures at the census tract level were assigned by linking geocoded maternal addresses from birth certificates to U.S. EPA National-Scale Air Toxics Assessment data for single and co-pollutant statistical analyses. Mixed-effects logistic regression models were applied to evaluate associations between air toxics and childhood leukemia.
In adjusted single pollutant models, odds of childhood leukemia among mothers with the highest ambient air exposures compared to those in the lowest quartile were 1.11 (95 % CI: 0.94-1.32) for POM, 1.17 (95 % CI: 0.98-1.39) for benzene and 1.29 (95 % CI: 1.08-1.52) for 1,3-butadiene. In co-pollutant models, odds ratios for childhood leukemia remained elevated for 1,3-butadiene but were close to the null value for benzene and POM.
We observed positive associations between 1,3-butadiene and childhood leukemia in single and co-pollutant models whereas effect estimates from single pollutant models were diminished for benzene and POM in co-pollutant models. Early life exposure to 1,3-butadiene rather than benzene or POM appears to increase early childhood risk of acute lymphocytic leukemia.

Isoprene is emitted in large quantities by vegetation, exhaled by human beings and released in small quantities by road traffic. As a result of its high reactivity, isoprene is an important ozone precursor in the troposphere and can play a key role in atmospheric chemistry. Measurements of isoprene in urban areas in Central Europe are scarce. Thus, in Essen, Germany, the isoprene concentration was measured at various sites during different seasons using two compact online GC-PID systems. Isoprene concentrations were compared with those of benzene and toluene, which represent typical anthropogenic VOCs. In the summer, the diurnal variation in isoprene concentration was dependent on the biogenic emissions in the city. It was found that its maximum concentration occurred during the day, in contrast to the benzene and toluene concentrations. During the measurement period in the summer of 2012, the average hourly isoprene concentrations reached 0.13 to 0.17 ppb between 10 and 20 LST. At high air temperatures, the isoprene concentration exceeded the benzene and toluene concentrations at many of the sites. Isoprene became more important than toluene with regard to ozone formation in the city area during the afternoon hours of summer days with high air temperatures. This finding was demonstrated by the contributions to OH reactivity and ozone-forming potential. It contradicts the results of other studies, which were based on daily or seasonal average values. With an isoprene/benzene ratio of 0.02, the contribution of anthropogenic isoprene decreased substantially to a very low level during the last 20 years in Central Europe due to a strong reduction in road traffic emissions. In the vicinity of many people, isoprene concentrations of up to 0.54 ppb and isoprene/benzene ratios of up to 1.34 were found in the atmosphere due to isoprene exhaled by humans.