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Transport derived Ultrafines and the Brain Effects

Periodic Reporting for period 3 - TUBE (Transport derived Ultrafines and the Brain Effects)

Reporting period: 2022-05-01 to 2023-10-31

In recent years, the indications showing that air pollution has effects beyond the lung are becoming more evident and neurological diseases, namely Alzheimer’s disease (AD) has shown to be associated with living near traffic. While the association of air pollutants with cognitive decline and neurodegenerative diseases has been discussed, the underlying molecular and cellular mechanisms and the components of air pollution that are responsible for these effects remain unknown. It is particularly important to understand the health impact of extremely small ultrafine particles (UFPs) as they readily cross bodily barriers and enter the brain, and as monitoring of UFPs is not mandatory and their emission limits are not regulated by legislation.

TUBE-project brings together interdisciplinary expertise to study the adverse effects of the extremely small UFPs in human lung and in the brain. It will focus on the effects of extremely small particles as well as (S)VOCs from combustion engines. TUBE-project will combine state of the art in vitro and in vivo models and epidemiological studies to disentangle the effects of transport-derived UFPs on health. The TUBE consortium has established a strong, multi-disciplinary, international group of experts to meet its objectives. The TUBE consortium is uniquely placed to decipher the effects of the extremely fine UFPs on biological processes leading to health impairment in the respiratory system and the brain to support planning a future traffic policy across the EU.

• Find out which emission sources or source environments cause the greatest exposure to UFPs.
• Recognize which components of air pollutants from transport are responsible for the adverse health effects seen in humans.
• Identify biomarkers and biological pathways.
• Understand the movement and clearance of particles in the brain.
• Resolve how UFPs affect brain health in humans.
• Provide mitigation strategies for emissions of road traffic and non-road equipment.
• Perform data integration and concomitant risk assessment.
• Provide data that will be used to support planning future EU policies.
We have optimized several methods and done sample collections for the testing of different emissions. Moreover, characterization of the emission environments and composition of the emissions has been started.
Cell-based methods have been optimized in several locations.

Sample collection (VTT, UEF):
VTT has collected several samples to be analysed in the project. We have now samples from current fleet technology with high aromatic and non-aromatic fuel. The same fuels have been collected for their semivolstile compounds.
In addition, UEF has participated in the same measurement campaigns with ALI system. This system was also used in previous measurement campaign at TAU, where VSParticle generator was used. VTT has also collected sample from the most recent technology (Euro 6d) in diesel car, and the collections will continue with other new technologies.

Cell experiments:
All partners involved, have been optimizing their cell culture models (UEF, UU, RIVM, IEM, MIM, BIOT).
This work is essential for the forthcoming experiments with exhaust samples. The consortium has also agreed to use the VSParticle generator samples as the reference for comparison between laboratories.
The fors experiments have been done by UEF (inhal tox), UU and UEF (neuro) with the collected material. UEF has done ALI exposures with various exhaust emissions as well.

In Vivo experiments:
Most of the in vivo experiments are planned later in the project. There has been, however, comtinuos discussion between the involved partners (UEF, Southampton, IUF) on planning the experiments and adjusting the research plan. University of Copenhagen will join in the second half of the project. In vivo experiments are planned for supproting the cell based experiments.

Human based approaches:
University of Umeå has started the experiments with healthy volunteers. However, this work has has slightly delayed due to Covid-19 situation. The analyses of the previous epidemiological results are on-going. The comparison of the results fomr parallel Chinese project will be conducted later, since the start of the project was somewhat delayed due to Covid. We will also receive the samples from China for analyses in toxicological experiments.

Air quality research:
The first measurement campaigns have been conducted, but the next ones had to be postponed due to Covid-19 situation. The measurement campaigns in Chile are being planned and they should be implemented next year.
We have developed the beyond state of the art approaches in all the areas of the project. The impact of these actions will be implemented in later stage of the project. Below the more detailed description of the WPs including scientific studies.

In WP1, the DDA system for separating small nanoparticles from the emissions for characterisation has been developed. We have also gained knowledge on the air pollution properties in different emission environments, including traffic environment in different locations, harbor environment, and airport location. The characterization has been done in comprehensive manner, using both stationary, and mobile laboratory. It has also been seen, how the latest emission technologies, fuels as well as DPF regeneration effects the emissions. The samples from the different cars operated with various aftertreatments and fuels are provided for analyses in WPs focusing on biological responses and health.

In WP2, the samples collected from WP1, including the clean car- campaign Euro 6d samples have been processed, and distributed to other partners, doing biological testing of the samples. Thus far, there has not been as extensive evaluation of different latest emission classification technologies as in TUBE. The emissions have also been analyzed using air-liquid interface exposure system, where also DDA was used in some of the experiments, as well as in different submerged cell cultures. WP2 has also performed a comparison study of three different ALI-systems, used in the project, to increase comparability between the results in different laboratories.

WP3 has continued the work on neuronal cell cultures, using the samples provided by WP1 and 2. The results have thus far revealed differences between fuels with different aromatic content, as well as difference between exhaust treatment technology. Extensive work on different cell models advances the research on air pollution effects to nervous system to a new level beyond state of the art.

In WP4, the experiments have been initiated to study the effect of engine exhaust exposure on clearance mechanisms from brain. The clearance pathways are utmost important in prevention of accumulation of harmful components to brain, which could then lead to e.g. AD development. Before this project, the information on these effects has been very limited. The work will be continuing by the analyses of the brain from exposed mice, and the work is on-going.

In WP5, COVID-19 pandemic delayed the work significantly, but now both the epidemiological and exposure work is on-going. This WP will give significant new information on the human effects of traffic-related air-pollutants.
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