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- How can multi-national occupational studies support policy making in Europe? Experiences from HBM4EU and PARC occupational studiesPublication . Santonen, Tiina; Louro, Henriqueta; Bocca, Beatrice; Bousoumah, Radia; Corneliu Duca, Radu; Fucic, Aleksandra; Galea, Karen S.; Godderis, Lode; Göen10, Thomas; Iavicoli, Ivo; Janasik, Beata; Jones, Kate; Leese, Elizabeth; Leso, Veruscka; Ndaw, Sophie; Poels, Katrien; Porras, Simo P.; Ruggieri, Flavia; Silva, Maria João; Van Nieuwenhuyse, An; Verdonck, Jelle; Wasowicz, Wojciech; Tavares, Ana; Sepai, Ovnair; Scheepers, Paul T.J.; Viegas, SusanaProviding policy relevant data on chemical exposures was a major aim of the EU biomonitor-ing initiative, HBM4EU. Similarly, the recently launched “Partnership for the risk assessment of chemicals” (PARC) emphasizes the relevance of the research findings to regulatory deci-sion making. Occupational biomonitoring studies performed under HBM4EU were focused on HBM4EU priority substances with specific regulatory relevance. Chromates and diisocya-nate studies were developed since these substances are of high concern for workers and to support regulatory measures under both EU chemicals regulation (REACH) and occupational safety and health (OSH) legislation. E-waste study was targeted to support European circular economy strategies and identify issues e.g. under OSH legislation. Based on the results of the chromates study, we drew several policy relevant conclusions which support the future up-dating of occupational exposure limits and the development of monitoring and risk man-agement practices in industry. A survey to policy makers on the usefulness of the study re-sults gave us confidence. We emphasise the importance of early and continued engagement of policy makers to ensure the usability of the results. Early communication with regulators of the anticipated benefits and formatting outputs appropriately are important and have been considered and improved within the EU PARC project.
- ToxApp4nanoCELFI – integrated view on the safety assessment of nanocellulosesPublication . Silva, Maria JoãoBackground: The expanding production and application of cellulose nanofibrils (CNF) has increased the risk of human exposure (workers and consumers); It is mandatory to ensure the safety of CNF for human health and the environment in a costeffective way – relevant to public and occupational health and also to industries; Because CNF are nanofibers with a high aspect ratio like MWCNT and crocidolite, the main question is whether some of the CNF synthesized may have deleterious effects on human health; Limited toxicological data is available for CNF given that most studies have focused on bacterial or nanocrystalline celulose; A previous study by the team showed that a CNF produced by TEMPO-mediated oxidation of an industrial bleached Eucalyptus globulus kraft pulp was genotoxic to alveolar cells (Ventura et al., 2018); The assessment of potential toxic effects of the newly synthesized CNF must be addressed in an early phase of the value chain to allow changes to be made before the production scale up (the safer-by-design approach).
- Overview of Adverse Outcome Pathways and Current Applications on NanomaterialsPublication . Rolo, Dora; Tavares, Ana; Vital, Nádia; Silva, Maria João; Louro, HenriquetaNanomaterials (NMs) have important and useful applications in chemical industry, electronics, pharmaceuticals, food and others. Their rapid proliferation presents a dilemma to regulators regarding hazard identification and increased concerns for public health. The Adverse Outcome Pathways (AOPs) are innovative central elements of a toxicological knowledge framework, developed for supporting chemical risk assessment based on mechanistic reasoning. AOPs describe a sequence of causally linked events at different levels of biological organisation, triggered by exposure to a stressor (like chemicals or NMs) leading to an adverse health effect in humans or wildlife. The integrative analysis of the cellular and molecular mechanisms of nanotoxicity towards the identification of connected adverse outcomes drives a sequential line – an AOP landscape definition. Each defined AOP is available for crossing data, linking known and unknown landscapes, reducing the reliance on animal studies, associated costs and ethical issues. NMs have unique properties, with specific associated toxicological challenges, which may represent unknown AOP landscapes. In this chapter, an overview of AOPs as important novel strategic tools in nanotoxicology is presented, highlighting the current applications in hazard identification and human health risk assessment.
- Cellular and Molecular Mechanisms of Toxicity of Ingested Titanium Dioxide NanomaterialsPublication . Vieira, Adriana; Gramacho, Ana; Rolo, Dora; Vital, Nádia; Silva, Maria João; Louro, HenriquetaAn exponential increase in products containing titanium dioxide nanomaterials (TiO2), in agriculture, food and feed industry, lead to increased oral exposure to these nanomaterials (NMs). Thus, the gastrointestinal tract (GIT) emerges as a possible route of exposure that may drive systemic exposure, if the intestinal barrier is surpassed. NMs have been suggested to produce adverse outcomes, such as genotoxic effects, that are associated with increased risk of cancer, leading to a concern for public health. However, to date, the differences in the physicochemical characteristics of the NMs studied and other variables in the test systems have generated contradictory results in the literature. Processes like human digestion may change the NMs characteristics, inducing unexpected toxic effects in the intestine. Using TiO2 as case-study, this chapter provides a review of the works addressing the interactions of NMs with biological systems in the context of intestinal tract and digestion processes, at cellular and molecular level. The knowledge gaps identified suggest that the incorporation of a simulated digestion process for in vitro studies has the potential to improve the model for elucidating key events elicited by these NMs, advancing the nanosafety studies towards the development of an adverse outcome pathway for intestinal effects.
- Risk assessment of combined occupational exposure to hexavalent chromium, nickel, and PAHs: a literature-based approachPublication . Tavares, Ana Maria; Viegas, Susana; Louro, Henriqueta; Silva, Maria JoãoOccupational exposure is usually characterized by a complex mixture of chemicals, originating from different raw materials and transformation processes. Co-exposure to hexavalent chromium (Cr(VI)), nickel (Ni) and polycyclic aromatic hydrocarbons (PAHs) can occur in some workplaces. These substances display well-known genotoxic and carcinogenic effects, especially in the respiratory tract, sharing similar modes of action. However, reference values for occupational exposure only account for individual components exposure and not for a potential mixture effect. In the scope of HBM4EU Initiative, we performed a mixtures risk assessment (MRA) based on literature from occupational studies conducted in the European Union that contain human biomonitoring (HBM) data on Cr(VI), Ni and/or PAHs. After HBM data extraction, Hazard Quotient (HQ) and Hazard Index (HI) were calculated for binary and tertiary mixtures. Exposure was considered of concern if HI>1. Twenty-four articles were selected, most (n=18, 75%) presenting Cr(VI) and Ni exposure biomarkers. Among these, HI>1 was obtained for all studies on welding activities, in which chromium was the main driver of toxicity with HQ>1 in most measurements. Only two studies in waste incineration setting reported exposure to the three substances, and again all HI>1. Noteworthy, for some of the analysed studies, although exposure levels were below the reference values, still the mixture was considered of concern (HI>1). Our findings show the limitations of applying occupational exposure reference values defined on a single substance basis to workplaces, highlighting the relevance of MRA as a more realistic approach to provide more suitable risk management measures in occupational settings.
- Investigation of potential respiratory adverse effects of micro/nanofibrillated cellulose and cellulose nanocrystals using human lung cell linesPublication . Pinto, Fátima; Louro, Henriqueta; Silva, Maria JoãoAbout cellulose nanomaterials (CNMs).
- Hazard Assessment of Benchmark Metal-Based Nanomaterials Through a Set of In Vitro Genotoxicity AssaysPublication . Vital, Nádia; Pinhão, Mariana; Yamani, Naouale El; Rundén-Pran, Elise; Louro, Henriqueta; Dušinská, Maria; Silva, Maria JoãoFor safety assessment of nanomaterials (NMs), in vitro genotoxicity data based on welldesigned experiments is required. Metal-based NMs are amongst the most used in consumer products. In this chapter, we report results for three metal-based NMs, titanium dioxide (NM- 100), cerium dioxide (NM-212) and silver (NM-302) in V79 cells, using a set of in vitro genotoxicity assays covering different endpoints: the medium-throughput comet assay and its modified version (with the enzyme formamidopyrimidine DNA glycosylase, Fpg), measuring DNA strand beaks (SBs) and oxidized purines, respectively; the micronucleus (MN) assay, assessing chromosomal damage; and the Hprt gene mutation test. The results generated by this test battery showed that all NMs displayed genotoxic potential. NM-100 induced DNA breaks, DNA oxidation damage and point mutations but not chromosome instability. NM-212 increased the level of DNA oxidation damage, point mutations and increased the MN frequency at the highest concentration tested. NM-302 was moderately cytotoxic and induced gene mutations, but not DNA or chromosome damage. In conclusion, the presented in vitro genotoxicity testing strategy allowed the identification of genotoxic effects caused by three different metal-based NMs, raising concern as to their impact on human health. The results support the use of this in vitro test battery for the genotoxicity assessment of NMs, reducing the use of more expensive, time-consuming and ethically demanding in vivo assays, in compliance with the 3 R’s.
- Exploring BPA alternatives - Environmental levels and toxicity reviewPublication . Adamovsky, Ondrej; Groh, Ksenia J.; Białk-Bielińska, Anna; Escher, Beate I.; Beaudouin, R.; Mora Lagares, Liadys; Tollefsen, Knut Erik; Fenske, Martina; Mulkiewicz, Ewa; Creusot, Nicolas; Sosnowska, Anita; Loureiro, Susana; Beyer, Jonny; Repetto, Guillermo; Štern, Alja; Lopes, Isabel; Monteiro, Marta; Zikova-Kloas, Andrea; Eleršek, Tina; Vračko, Marjan; Zdybel, Szymon; Puzyn, Tomasz; Koczur, Weronika; Ebsen Morthorst, Jane; Holbech, Henrik; Carlsson, Gunnar; Örn, Stefan; Herrero, Óscar; Siddique, Ayesha; Liess, Matthias; Braun, Georg; Srebny, Vanessa; Žegura, Bojana; Hinfray, Nathalie; Brion, François; Knapen, Dries; Vandeputte, Ellen; Stinckens, Evelyn; Vergauwen, Lucia; Behrendt, Lars; Silva, Maria João; Blaha, Ludek; Kyriakopoulou, KaterinaBisphenol A alternatives are manufactured as potentially less harmful substitutes of bisphenol A (BPA) that offer similar functionality. These alternatives are already in the market, entering the environment and thus raising ecological concerns. However, it can be expected that levels of BPA alternatives will dominate in the future, they are limited information on their environmental safety. The EU PARC project highlights BPA alternatives as priority chemicals and consolidates information on BPA alternatives, with a focus on environmental relevance and on the identification of the research gaps. The review highlighted aspects and future perspectives. In brief, an extension of environmental monitoring is crucial, extending it to cover BPA alternatives to track their levels and facilitate the timely implementation of mitigation measures. The biological activity has been studied for BPA alternatives, but in a non-systematic way and prioritized a limited number of chemicals. For several BPA alternatives, the data has already provided substantial evidence regarding their potential harm to the environment. We stress the importance of conducting more comprehensive assessments that go beyond the traditional reproductive studies and focus on overlooked relevant endpoints. Future research should also consider mixture effects, realistic environmental concentrations, and the long-term consequences on biota and ecosystems.
- SOP3 Blood Sampling - Partnership for the Assessment of the Risks from ChemicalsPublication . Silva, Maria João; Henriqueta LouroSOP3 Blood Sampling - presentation for PARC Training for Waste Management Survey. Aim of the presentation: Training dedicated to introduce contents of SOP for blood sampling, transport and storage, to ensure its correct implementation during the waste management occupational study. This Standard Operating Procedure (SOP) for blood sampling provides the general procedure for the collection, storage and transfer of human blood samples to be analysed within the e-waste occupational study under HBM4EU and is based on a SOP developed within a previous occupational study.
- SOP9 Buccal Cells Sampling - Partnership for the Assessment of the Risks from ChemicalsPublication . Louro, Henriqueta; Silva, Maria JoãoSOP9 Buccal Cells Sampling - presentation for PARC Training for Waste Management Survey / Partnership for the Assessment of the Risks from Chemicals. Aim of presentation: Provide the general procedure for the collection, storage, and transfer of buccal cell samples for micronucleus analysis in exfoliated buccal cells. The buccal micronucleus assay is a minimally invasive approach for measuring DNA damage, cell proliferation, cell differentiation and cell death in exfoliated buccal cells (Bolognesi et al., 2015). It offers a great opportunity to evaluate in a clear and precise way the appearance of genetic damage whether it is present as a consequence of occupational or environmental risk, being reliable, fast, relatively simple, cheap, and minimally invasive and causes no pain. This Standard Operating Procedure (SOP) for buccal cells sampling is intended to be used in the PARC waste management survey. The SOP provides the general procedure for the collection, storage, and transfer of buccal cell samples.