This study used toxicogenomics to identify the complex biological response of human lung BEAS-2B cells treated with organic components of particulate matter in the exhaust of a diesel engine. First, we characterized particles from standard diesel (B0), biodiesel (methylesters of rapeseed oil) in its neat form (B100) and 30% by volume blend with diesel fuel (B30), and neat hydrotreated vegetable oil (NEXBTL100). The concentration of polycyclic aromatic hydrocarbons (PAHs) and their derivatives in organic extracts was the lowest for NEXBTL100 and higher for biodiesel. We further analyzed global gene expression changes in BEAS-2B cells following 4 h and 24 h treatment with extracts. The concentrations of 50 µg extract/mL induced a similar molecular response. The common processes induced after 4 h treatment included antioxidant defense, metabolism of xenobiotics and lipids, suppression of pro-apoptotic stimuli, or induction of plasminogen activating cascade; 24 h treatment affected fewer processes, particularly those involved in detoxification of xenobiotics, including PAHs. The majority of distinctively deregulated genes detected after both 4 h and 24 h treatment were induced by NEXBTL100; the deregulated genes included, e.g., those involved in antioxidant defense and cell cycle regulation and proliferation. B100 extract, with the highest PAH concentrations, additionally affected several cell cycle regulatory genes and p38 signaling.

Center of Vehicles for Sustainable Mobility Faculty of Mechanical Engineering Czech Technical University Prague Technicka 4 166 07 Prague Czech Republic

Department of Chemistry and Toxicology Veterinary Research Institute Hudcova 296 70 621 00 Brno Czech Republic

Department of Cybernetics Faculty of Electrical Engineering Czech Technical University Prague Karlovo namesti 13 121 35 Prague Czech Republic

Department of Genetic Toxicology and Nanotoxicology Institute of Experimental Medicine AS CR Videnska 1083 142 20 Prague Czech Republic

Department of Genetic Toxicology and Nanotoxicology Institute of Experimental Medicine AS CR Videnska 1083 142 20 Prague Czech Republic Faculty of Safety Engineering VSB Technical University of Ostrava Lumirova 13 700 30 Ostrava Czech Republic

Department of Genetic Toxicology and Nanotoxicology Institute of Experimental Medicine AS CR Videnska 1083 142 20 Prague Czech Republic Institute for Environmental Studies Faculty of Science Charles University Prague Benatska 2 128 01 Prague 2 Czech Republic

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$a Libalova, Helena $u Department of Genetic Toxicology and Nanotoxicology, Institute of Experimental Medicine AS CR, Videnska 1083, 142 20 Prague, Czech Republic. libalova@biomed.cas.cz.

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$a Comparative Analysis of Toxic Responses of Organic Extracts from Diesel and Selected Alternative Fuels Engine Emissions in Human Lung BEAS-2B Cells / $c H. Libalova, P. Rossner, K. Vrbova, T. Brzicova, J. Sikorova, M. Vojtisek-Lom, V. Beranek, J. Klema, M. Ciganek, J. Neca, K. Pencikova, M. Machala, J. Topinka,

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$a This study used toxicogenomics to identify the complex biological response of human lung BEAS-2B cells treated with organic components of particulate matter in the exhaust of a diesel engine. First, we characterized particles from standard diesel (B0), biodiesel (methylesters of rapeseed oil) in its neat form (B100) and 30% by volume blend with diesel fuel (B30), and neat hydrotreated vegetable oil (NEXBTL100). The concentration of polycyclic aromatic hydrocarbons (PAHs) and their derivatives in organic extracts was the lowest for NEXBTL100 and higher for biodiesel. We further analyzed global gene expression changes in BEAS-2B cells following 4 h and 24 h treatment with extracts. The concentrations of 50 µg extract/mL induced a similar molecular response. The common processes induced after 4 h treatment included antioxidant defense, metabolism of xenobiotics and lipids, suppression of pro-apoptotic stimuli, or induction of plasminogen activating cascade; 24 h treatment affected fewer processes, particularly those involved in detoxification of xenobiotics, including PAHs. The majority of distinctively deregulated genes detected after both 4 h and 24 h treatment were induced by NEXBTL100; the deregulated genes included, e.g., those involved in antioxidant defense and cell cycle regulation and proliferation. B100 extract, with the highest PAH concentrations, additionally affected several cell cycle regulatory genes and p38 signaling.

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$a Brzicova, Tana $u Department of Genetic Toxicology and Nanotoxicology, Institute of Experimental Medicine AS CR, Videnska 1083, 142 20 Prague, Czech Republic. tana.brzicova@biomed.cas.cz. Faculty of Safety Engineering, VSB-Technical University of Ostrava, Lumirova 13, 700 30 Ostrava, Czech Republic. tana.brzicova@biomed.cas.cz.

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$a Vojtisek-Lom, Michal $u Center of Vehicles for Sustainable Mobility, Faculty of Mechanical Engineering, Czech Technical University in Prague, Technicka 4, 166 07 Prague, Czech Republic. michal.vojtisek@fs.cvut.cz.

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