Ordinary Gasoline Emissions Induce a Toxic Response in Bronchial Cells Grown at Air-Liquid Interface.

Gasoline engine emissions have been classified as possibly carcinogenic to humans and represent a significant health risk. In this study, we used MucilAir™, a three-dimensional (3D) model of the human airway, and BEAS-2B, cells originating from the human bronchial epithelium, grown at the air-liquid interface to assess the toxicity of ordinary gasoline exhaust produced by a direct injection spark ignition engine. The transepithelial electrical resistance (TEER), production of mucin, and lactate dehydrogenase (LDH) and adenylate kinase (AK) activities were analyzed after one day and five days of exposure. The induction of double-stranded DNA breaks was measured by the detection of histone H2AX phosphorylation. Next-generation sequencing was used to analyze the modulation of expression of the relevant 370 genes. The exposure to gasoline emissions affected the integrity, as well as LDH and AK leakage in the 3D model, particularly after longer exposure periods. Mucin production was mostly decreased with the exception of longer BEAS-2B treatment, for which a significant increase was detected. DNA damage was detected after five days of exposure in the 3D model, but not in BEAS-2B cells. The expression of CYP1A1 and GSTA3 was modulated in MucilAir™ tissues after 5 days of treatment. In BEAS-2B cells, the expression of 39 mRNAs was affected after short exposure, most of them were upregulated. The five days of exposure modulated the expression of 11 genes in this cell line. In conclusion, the ordinary gasoline emissions induced a toxic response in MucilAir™. In BEAS-2B cells, the biological response was less pronounced, mostly limited to gene expression changes.

The Biological Effects of Complete Gasoline Engine Emissions Exposure in a 3D Human Airway Model (MucilAirTM) and in Human Bronchial Epithelial Cells (BEAS-2B).

The biological effects induced by complete engine emissions in a 3D model of the human airway (MucilAirTM) and in human bronchial epithelial cells (BEAS-2B) grown at the air-liquid interface were compared. The cells were exposed for one or five days to emissions generated by a Euro 5 direct injection spark ignition engine. The general condition of the cells was assessed by the measurement of transepithelial electrical resistance and mucin production. The cytotoxic effects were evaluated by adenylate kinase (AK) and lactate dehydrogenase (LDH) activity. Phosphorylation of histone H2AX was used to detect double-stranded DNA breaks. The expression of the selected 370 relevant genes was analyzed using next-generation sequencing. The exposure had minimal effects on integrity and AK leakage in both cell models. LDH activity and mucin production in BEAS-2B cells significantly increased after longer exposures; DNA breaks were also detected. The exposure affected CYP1A1 and HSPA5 expression in MucilAirTM. There were no effects of this kind observed in BEAS-2B cells; in this system gene expression was rather affected by the time of treatment. The type of cell model was the most important factor modulating gene expression. In summary, the biological effects of complete emissions exposure were weak. In the specific conditions used in this study, the effects observed in BEAS-2B cells were induced by the exposure protocol rather than by emissions and thus this cell line seems to be less suitable for analyses of longer treatment than the 3D model.

Markers of lipid oxidation and inflammation in bronchial cells exposed to complete gasoline emissions and their organic extracts.

Road traffic emissions consist of gaseous components, particles of various sizes, and chemical compounds that are bound to them. Exposure to vehicle emissions is implicated in the etiology of inflammatory respiratory disorders. We investigated the inflammation-related markers in human bronchial epithelial cells (BEAS-2B) and a 3D model of the human airways (MucilAir™), after exposure to complete emissions and extractable organic matter (EOM) from particles generated by ordinary gasoline (E5), and a gasoline-ethanol blend (E20; ethanol content 20% v/v). The production of 22 lipid oxidation products (derivatives of linoleic and arachidonic acid, AA) and 45 inflammatory molecules (cytokines, chemokines, growth factors) was assessed after days 1 and 5 of exposure, using LC-MS/MS and a multiplex immunoassay, respectively. The response observed in MucilAir™ exposed to E5 gasoline emissions, characterized by elevated levels of pro-inflammatory AA metabolites (prostaglandins) and inflammatory markers, was the most pronounced. E20 EOM exposure was associated with increased levels of AA metabolites with anti-inflammatory effects in this cell model. The exposure of BEAS-2B cells to complete emissions reduced lipid oxidation, while E20 EOM tended to increase concentrations of AA metabolite and chemokine production; the impacts on other inflammatory markers were limited. In summary, complete E5 emission exposure of MucilAir™ induces the processes associated with the pro-inflammatory response. This observation highlights the potential negative health impacts of ordinary gasoline, while the effects of alternative fuel are relatively weak.

On-road detection of trucks with high NOx emissions from a patrol vehicle with on-board FTIR analyzer.

Technological advances in heavy-duty vehicle engines, allowing them to reach NOx emissions comparable to European diesel passenger cars per km driven, are being compromised by aftermarket defeat devices such as selective catalytic reduction (SCR) emulators, many of which can be quickly deactivated by the driver. In a pilot study, the prevalence of trucks with excess NOx emissions on Czech motorways was evaluated using an ordinary Customs Administration patrol vehicle temporarily fitted with a portable fast-response Fourier Transform Infra Red (FTIR) analyzer, acting as an impromptu chase vehicle. The Euro emissions category of the truck was provided from the motorway toll collection transponders. A total of 222 unique trucks were measured during a one-week pilot project. Of these, 66% were Euro VI, 25% were Euro V, and 9% were older categories. NO/CO2 ratios were calculated as a ratio of numerical integrals of the peaks of measured concentrations, as a ratio of maximum measured concentrations, and by linear regression, with the regression approach yielding most realistic results and mean calculated error of 0.2 g/kWh NO. At assumed 85% NO in NOx and 634 g/kWh mean CO2 emissions, the mean emissions of the cleanest 83% of Euro V and cleanest 63% of Euro VI trucks were within the corresponding NOx limit (2 g/kWh for Euro V, 0.46 g/kWh for Euro VI) multiplied by a factor of 1.5. Providing for some allowance for legitimate occurrences of high NOx emissions, about 10-15% of Euro V and about 10-25% of Euro VI trucks are believed to be excess emitters, with no SCR functionality on about 10-15% of Euro VI trucks. The portable FTIR, temporarily mounted on a law enforcement vehicle, can be readily used as a screening tool, identifying vehicles to be stopped for additional inspection, but also during roadside emissions inspections.

Comparison of hydrogenated vegetable oil and biodiesel effects on combustion, unregulated and regulated gaseous pollutants and DPF regeneration procedure in a Euro6 car.

The effects of traditional biodiesel (fatty acid methyl-esters, FAME) and a hydrotreated vegetable oil (HVO) were comprehensively investigated on a production Euro 6 diesel car, including fuel injection rate and timing, combustion analysis, emissions of regulated and unregulated pollutants, and regeneration of the diesel particle filter. The use of both biofuels is a part of the efforts to reduce emissions of greenhouse gases and health-relevant pollutants and to improve energy security and sustainability. HVO, albeit more expensive, offers benefits relative to FAME in terms of oxidation stability, injector fouling, energy content and cetane number. The car was fitted with an on-board instrumentation and subjected to a range of driving cycles on a chassis dynamometer. The fuel consumption calculated from instantaneous emissions data based on exhaust gas composition measured by an on-board FTIR and calculated exhaust flow matched directly measured fuel consumption within several percent on all fuels; differences in the consumption among the fuels correspond to different heating values. The combustion onset and maximum heat release rate were comparable for diesel and FAME but were advanced on HVO due to its higher cetane number, causing, at times, multiple distinct heat release peaks, suggesting that optimization of fuel injection timing for HVO might be beneficial. Emissions of methane and ammonia were negligible, of N2O were measurable and slightly lower for HVO than for other fuels, of formaldehyde were limited to cold engine accelerations and highest for FAME and negligible for HVO, of NO and NO2 were high on all fuels during all operating conditions except for the type approval test. The results confirm several relative advantages of HVO over RME, with penetration into engine lubricating oil during particle filter regeneration to be further investigated. The effects of HVO lubricity and other long-term effects were not evaluated here.