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      Particulate Matter from Both Heavy Fuel Oil and Diesel Fuel Shipping Emissions Show Strong Biological Effects on Human Lung Cells at Realistic and Comparable In Vitro Exposure Conditions

      research-article
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          Abstract

          Background

          Ship engine emissions are important with regard to lung and cardiovascular diseases especially in coastal regions worldwide. Known cellular responses to combustion particles include oxidative stress and inflammatory signalling.

          Objectives

          To provide a molecular link between the chemical and physical characteristics of ship emission particles and the cellular responses they elicit and to identify potentially harmful fractions in shipping emission aerosols.

          Methods

          Through an air-liquid interface exposure system, we exposed human lung cells under realistic in vitro conditions to exhaust fumes from a ship engine running on either common heavy fuel oil (HFO) or cleaner-burning diesel fuel (DF). Advanced chemical analyses of the exhaust aerosols were combined with transcriptional, proteomic and metabolomic profiling including isotope labelling methods to characterise the lung cell responses.

          Results

          The HFO emissions contained high concentrations of toxic compounds such as metals and polycyclic aromatic hydrocarbon, and were higher in particle mass. These compounds were lower in DF emissions, which in turn had higher concentrations of elemental carbon (“soot”). Common cellular reactions included cellular stress responses and endocytosis. Reactions to HFO emissions were dominated by oxidative stress and inflammatory responses, whereas DF emissions induced generally a broader biological response than HFO emissions and affected essential cellular pathways such as energy metabolism, protein synthesis, and chromatin modification.

          Conclusions

          Despite a lower content of known toxic compounds, combustion particles from the clean shipping fuel DF influenced several essential pathways of lung cell metabolism more strongly than particles from the unrefined fuel HFO. This might be attributable to a higher soot content in DF. Thus the role of diesel soot, which is a known carcinogen in acute air pollution-induced health effects should be further investigated. For the use of HFO and DF we recommend a reduction of carbonaceous soot in the ship emissions by implementation of filtration devices.

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          Most cited references29

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          Evidence on the impact of sustained exposure to air pollution on life expectancy from China's Huai River policy.

          This paper's findings suggest that an arbitrary Chinese policy that greatly increases total suspended particulates (TSPs) air pollution is causing the 500 million residents of Northern China to lose more than 2.5 billion life years of life expectancy. The quasi-experimental empirical approach is based on China's Huai River policy, which provided free winter heating via the provision of coal for boilers in cities north of the Huai River but denied heat to the south. Using a regression discontinuity design based on distance from the Huai River, we find that ambient concentrations of TSPs are about 184 μg/m(3) [95% confidence interval (CI): 61, 307] or 55% higher in the north. Further, the results indicate that life expectancies are about 5.5 y (95% CI: 0.8, 10.2) lower in the north owing to an increased incidence of cardiorespiratory mortality. More generally, the analysis suggests that long-term exposure to an additional 100 μg/m(3) of TSPs is associated with a reduction in life expectancy at birth of about 3.0 y (95% CI: 0.4, 5.6).
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            Mortality from ship emissions: a global assessment.

            Epidemiological studies consistently link ambient concentrations of particulate matter (PM) to negative health impacts, including asthma, heart attacks, hospital admissions, and premature mortality. We model ambient PM concentrations from oceangoing ships using two geospatial emissions inventories and two global aerosol models. We estimate global and regional mortalities by applying ambient PM increases due to ships to cardiopulmonary and lung cancer concentration-risk functions and population models. Our results indicate that shipping-related PM emissions are responsible for approximately 60,000 cardiopulmonary and lung cancer deaths annually, with most deaths occurring near coastlines in Europe, East Asia, and South Asia. Under current regulation and with the expected growth in shipping activity, we estimate that annual mortalities could increase by 40% by 2012.
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              Transformation of human bronchial epithelial cells by infection with SV40 or adenovirus-12 SV40 hybrid virus, or transfection via strontium phosphate coprecipitation with a plasmid containing SV40 early region genes.

              Normal human bronchial epithelial cells were infected with SV40 virus or an adenovirus 12-SV40 hybrid virus, or transfected via strontium phosphate coprecipitation with plasmids containing the SV40 early region genes. Colonies of morphologically altered cells were isolated and cultured; these cells had extended culture lifespans compared to normal human bronchial epithelial cells. All cultures eventually underwent senescence, with the exception of one which appears to have unlimited proliferative potential. Colonies arising after viral infection were screened for virus production by cocultivation with Vero cells; only viral nonproducer cultures were analyzed further. The cells retained electron microscopic features of epithelial cells, and keratin and SV40 T-antigen were detected by indirect immunofluorescence. All of the cultures were aneuploid with karyotypic abnormalities characteristic of SV40-transformed cells. No tumors formed after s.c. injection of the cells in nude mice. These cells should be useful for studies of multistage bronchial epithelial carcinogenesis.
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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                3 June 2015
                2015
                : 10
                : 6
                : e0126536
                Affiliations
                [1 ]HICE—Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health—Aerosols and Health, www.hice-vi.eu, Neuherberg, Rostock, Munich, Karlsruhe, Berlin, Waldkirch, Germany, Kuopio, Finland, Cardiff, UK, Esch-Belval, Luxembourg
                [2 ]Center of Allergy and Environment (ZAUM), Helmholtz Zentrum München and Technische Universität München, Member of the German Center for Lung Research (DZL), Munich, Germany
                [3 ]CK-CARE, Christine Kühne Center for Allergy Research and Education, Davos, Switzerland
                [4 ]Mass Spectrometry Core Unit, Max Delbrück Center for Molecular Medicine Berlin-Buch, Germany
                [5 ]University of Eastern Finland, Department of Environmental Science, P.O. Box 1627, FI-70211 Kuopio, Finland
                [6 ]Luxembourg Centre for Systems Biomedicine, University of Luxembourg, L-4362 Esch-Belval, Luxembourg
                [7 ]Joint Mass Spectrometry Centre, Chair of Analytical Chemistry, Institute of Chemistry, University Rostock, Rostock, Germany
                [8 ]Joint Mass Spectrometry Centre, CMA—Comprehensive Molecular Analytics, Helmholtz Zentrum München, Neuherberg, Germany
                [9 ]Institute for Technical Chemistry (ITC), Karlsruhe Institute of Technology, Campus North, Karlsruhe, Germany
                [10 ]Institute of Toxicology and Genetics (ITG), Karlsruhe Institute of Technology, Campus North, Karlsruhe, Germany
                [11 ]Chair of Piston Machines and Internal Combustion Engines, University Rostock, Rostock, Germany
                [12 ]VTT Technical Research Centre of Finland, P.O. Box 1000, FI-02044 VTT, Espoo, Finland
                [13 ]National Institute for Health and Welfare, Department of Environmental Health, P.O. Box 95, FI-70701, Kuopio, Finland
                [14 ]Institute of environmental medicine, UNIKA-T, Technische Universität, Munich, Germany
                [15 ]Lung and Particle Research Group, School of Biosciences, Cardiff University, Cardiff, Wales, United Kingdom
                [16 ]Research Unit Analytical BioGeoChemistry, Helmholtz Zentrum München—German Research Center for Environmental Health GmbH, Neuherberg, Germany
                [17 ]Vitrocell GmbH, Waldkirch, Germany
                [18 ]Laboratory of Atmospheric Chemistry, Paul Scherrer Institute (PSI), Villigen, Switzerland
                [19 ]Institute of Physics, University Rostock, Rostock, Germany
                University of Alabama at Birmingham, UNITED STATES
                Author notes

                Competing Interests: Tobias Krebs is an employee of Vitrocell GmbH, Tübingen, Germany. This does not alter the authors' adherence to PLOS ONE policies on sharing data and materials.

                Conceived and designed the experiments: SO T. Kanashova OS SCS T. Streibel JP MD HRP SM SD CW HH JKJ MRH KAB MK EK GJ MS JO LM ME AR TG CR KH JB GD RZ. Performed the experiments: SO T. Kanashova OS SCS JP MD CS BS RR TT AJW ZP BM AP MK EK GJ JL GM MS JO PR LM ME AR BW T. Schwemer HC CPR GA CR. Analyzed the data: SO T. Kanashova OS SCS T. Streibel JMAS JP MD SD CW BS RR TT AJW ZP BM AP MK JT EK GJ JSK JL GM MS SK JO PR LM ME AR TG BW T. Schwemer HC CPR GA CR KH JB GD RZ. Contributed reagents/materials/analysis tools: SO T. Kanashova OS SCS T. Streibel JMAS JP MD HRP CS SM SD CW BS RR HH TT JKJ MRH CSW CTH KAB AJW ZP BM T. Krebs AP MK JT EK GJ SS JSK JL GM MS SK JO PR LM ME AR TG BW T. Schwemer HC CPR GA CR KH JB GD RZ. Wrote the paper: SO T. Kanashova OS SCS T. Streibel JMAS JP MD SM BS KAB AJW ZP JT EK GJ SS JSK JL GM MS SK JO LM ME AR TG BW T. Schwemer HC CPR GA CR KH JB GD RZ. Initially conceived and designed the study: T. Streibel HRP CW HH JKJ MRH KAB T. Krebs JT EK JSK JL KM MS TG KH JB GD RZ.

                ‡ These authors also contributed equally to the manuscript.

                Article
                PONE-D-14-45336
                10.1371/journal.pone.0126536
                4454644
                26039251
                bd427413-dcb0-4c48-8b05-dc409fa1329d
                Copyright @ 2015

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

                History
                : 14 October 2014
                : 2 April 2015
                Page count
                Figures: 3, Tables: 1, Pages: 17
                Funding
                HICE partners received funding from the Impulse and Networking Funds (INF) of the Helmholtz Association (HGF), Berlin, Germany. The support of HICE by the Helmholtz Zentrum München and University of Rostock is gratefully acknowledged. Sebastian Oeder also received funding from CK-CARE Teilbereich A. Sean Sapcariu and Karsten Hiller acknowledge financial support from the Fonds National de la Recherche (FNR), specifically the ATTRACT program Metabolomics Junior Group. Funding from the Academy of Finland (Grant No: 258315 & 259946), Saastamoinen foundation and the strategic funding of the University of Eastern Finland for project “sustainable bioenergy, climate change and health” is acknowledged. Funding from the German Science Foundation (DFG ZI 764/5-1, ZI 764/3-1, INST 264/56-1 and 264/77-1) helped to achieve the presented results. We also thank SNSF and DFG for funding for the DACH project WOOSHI. Vitrocell GmbH provided support in the form of a salary for author T. Krebs, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of the authors are articulated in the ‘author contributions’ section.
                Categories
                Research Article
                Custom metadata
                All relevant data are within the paper and its Supporting Information files. Additionally, transcriptomics data are available from Gene Expression Omnibus (accession number GSE63962). Proteomics data are available from ProteomicsDB (ID: PRDB004215).

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