Risk-benefit assessment of shifting from traditional meat-based diets to alternative dietary patterns.

The consumption of plant-based meat alternatives has recently transitioned from the niche market to the mainstream. However, changing the dietary patterns may lead to new health challenges referred to possible higher exposure to natural toxins found in plant-based food. The present project aimed to train the fellow in the field of chemical risk assessment and provide a comprehensive overview on how a change towards plant-based meat alternatives may represent a driver for emerging risks. Thus, within the EU-FORA programme the fellow engaged in the following activities: (i) perform a systematic review to analyse the distribution of natural toxins in the most common plant-based meat alternatives in Europe, (ii) risk-benefit assessment of shifting from meat-based diets to soy-based meat alternatives by modelling the substitution of meat with soy, and (iii) determine the occurrence of mycotoxins in plant-based meat alternatives from local markets in Parma, Italy. The fellow learnt the chemical risk assessment procedures applied by the researchers from the Department of Food and Drug of Parma University, thus gaining an in-depth expertise in all the steps. A risk-benefit assessment was performed modelling the intake of aflatoxin B1-contaminated soy-based meat analogues. The health impact due to intake of soy and exposure to aflatoxin B1 was estimated. Within the research group, the fellow also worked on developing a multi-mycotoxin determination method for plant-based meat alternatives matrices. The results of the project provide a picture reflecting the occurrence of natural toxins in plant-based meat alternatives and the need of upgraded regulation frameworks that take into account new products and dietary patterns. The EU-FORA fellowship was a great opportunity for the fellow to expand his professional network and increase his expertise in food safety by gaining new skills in chemical risk assessment, risk-benefit assessment and analytical chemistry.

Assessment of the Multifunctional Behavior of Lupin Peptide P7 and Its Metabolite Using an Integrated Strategy.

LTFPGSAED (P7) is a multifunctional hypocholesterolemic and hypoglycemic lupin peptide. While assessing its angiotensin-converting enzyme (ACE) inhibitory activity, it was more effective in intestinal Caco-2 cells (IC50 of 13.7 µM) than in renal HK-2 cells (IC50 of 79.6 µM). This discrepancy was explained by the metabolic transformation mediated by intestinal peptidases, which produced two main detected peptides, TFPGSAED and LTFPG. Indeed LTFPG, dynamically generated by intestinal dipeptidyl peptidase IV as well as its parent peptide P7 were linearly absorbed by mature Caco-2 cells. An in silico study demonstrated that the metabolite was a better ligand of the ACE enzyme than P7. These results are in agreement with an in vivo study, previously performed by Aluko et al., which has shown that LTFPG is an effective hypotensive peptide. Our work highlights the dynamic nature of bioactive food peptides that may be modulated by the metabolic activity of intestinal cells.

Toxicodynamics of Mycotoxins in the Framework of Food Risk Assessment-An In Silico Perspective.

Mycotoxins severely threaten the health of humans and animals. For this reason, many countries have enforced regulations and recommendations to reduce the dietary exposure. However, even though regulatory actions must be based on solid scientific knowledge, many aspects of their toxicological activity are still poorly understood. In particular, deepening knowledge on the primal molecular events triggering the toxic stimulus may be relevant to better understand the mechanisms of action of mycotoxins. The present work presents the use of in silico approaches in studying the mycotoxins toxicodynamics, and discusses how they may contribute in widening the background of knowledge. A particular emphasis has been posed on the methods accounting the molecular initiating events of toxic action. In more details, the key concepts and challenges of mycotoxins toxicology have been introduced. Then, topical case studies have been presented and some possible practical implementations of studying mycotoxins toxicodynamics have been discussed.

Hazard assessment through hybrid in vitro / in silico approach: The case of zearalenone.

Within the framework of reduction, refinement and replacement of animal experiments, new approaches for identification and characterization of chemical hazards have been developed. Grouping and read across has been promoted as a most promising alternative approach. It uses existing toxicological information on a group of chemicals to make predictions on the toxicity of uncharacterized ones. In the present work, the feasibility of applying in vitro and in silico techniques to group chemicals for read across was studied using the food mycotoxin zearalenone (ZEN) and metabolites as a case study. ZEN and its reduced metabolites are known to act through activation of the estrogen receptor α (ERα). The ranking of their estrogenic potencies appeared highly conserved across test systems including binding, in vitro and in vivo assays. This data suggests that activation of ERα may play a role in the molecular initiating event (MIE) and be predictive of adverse effects and provides the rationale to model receptor-binding for hazard identification. The investigation of receptor-ligand interactions through docking simulation proved to accurately rank estrogenic potencies of ZEN and reduced metabolites, showing the suitability of the model to address estrogenic potency for this group of compounds. Therefore, the model was further applied to biologically uncharacterized, commercially unavailable, oxidized ZEN metabolites (6α-, 6ß-, 8α-, 8ß-, 13- and 15-OH-ZEN). Except for 15-OH-ZEN, the data indicate that in general, the oxidized metabolites would be considered a lower estrogenic concern than ZEN and reduced metabolites.