Methodology & structure
PATHWAY-27 strategy and methodology is based on 8 tightly connected workpackages, whose leaders have been selected for experience in transnational research activities, internationally recognized scientific knowledge in the area concerned, effective communication skills and enthusiasm.
WP1 Coordination and Management (leader Alessandra Bordoni - UNIBO)
The main objectives of WP1 are:
• Coordinate and supervise the research activities between Work Packages, monitor the overall Project achievements and control the scientific quality and timing of the Project results;
• Manage contacts with EU Commission and among the consortium;
• Implement the EU Grant Agreement and the Consortium Agreement;
• Supervise the foreground generated by the project activities monitoring that IP is properly managed;
• Ensure an appropriate overall financial and administrative management and reporting;
• Ethics management.
WP2 Feasibility of selected food matrices for the formulation of PATHWAY-27 bioactive-enriched foods (leader: Didier Dupont –INRA)
The overall objective of WP2 is to formulate 9 BEF enriched with DHA, BG, AC alone, or DHA in combination with BG, or DHA in combination with AC possessing optimised bioactive accessibility and sensory properties.
Three different matrices will be used for the development of BEF:
i) bakery products, BP;
ii) dairy products, DP;
iii) egg products, EP.
The interaction of the bioactive with the food matrix, together with the protective effect of the food microstructure on the resistance of bioactives during gastrointestinal tract transit, will determine the selection of optimized BEF. SMEs will be heavily involved in the formulation, design and production of the different BEF which, where necessary, will include stabilizers and protectives for bioactives, as well as balancing sensory properties. The stability of the bioactives and their effect on the food matrix at the end of the food production cycle, storage and shelf-life of the product will be determined. State-of-the-art food-omics techniques, such as metabonomics for the molecular profiling, and low field NMR for food matrix structure characterization, will be used to gain the most complete food description of the enriched and non-enriched foods.
The appropriate formulation for the correct inclusion of the 5 combinations of bioactive compounds in the 9 food products will be reached by factorial designs to simultaneously evaluate the effect of the following variables for product preparation: ingredients, stabilizers, salt concentration, pH, processing parameters like mixing time/speed, other processing times and temperatures, etc. Particular attention will be devoted to: the optimization of the ratio of bioactives, proteins, fats and other ingredients; the choice of suitable ionic strength/pH parameters to control the aggregation of hydrophilic macromolecules in order to obtain discrete colloidal particles; the selection of suitable excipients in order to enable equilibration of components to achieve colloidal particles; the selection of suitable stabilizers to control "host-guest" interaction between polysaccharides and AC. Particular emphasis will also be placed on studying the dissociation of colloidal structures at varying pH, ionic strength and enzymatic action in order to modulate the release of natural actives in chemically unaltered forms.
WP3 In vitro studies on the effects and mechanism of action of selected bioactives (leader: Lidia Tomas – AINIA)
The main aim of WP3 is to understand the protective role and mechanism of action of each bioactive compound (DHA, BG and AC), their interaction at the cellular level and their role in the aetiology and development of the Metabolic Syndrome (MS). WP3 will clarify the function and the compatibility of the combinations, investigating their possible synergistic, antagonistic or neutral effects.
Specific objectives are:
1. To study and to evaluate the impact of DHA, alone and in combination with BG or AC metabolites, on transcriptional and epigenetic regulation and on cellular metabolism and insulin resistance.
2. To provide scientific evidence for the health benefits of DHA, alone and in combination with BG or AC.
Results obtained will be used to select specific biomarkers to be measured in WP6 on volunteers recruited in the intervention study. Additionally, results obtained in WP6 will feed-back to WP3 for in-depth mechanistic analyses in vitro.
WP4 Industrial production of PATHWAY BEF for pilot and intervention studies (leader: Laura Flander – VTT)
The overall objective of WP4 is to extract and purify the selected bioactives from dietary sources, and to use them for the production of bioactive-enriched foods (BEF) for the pilot and intervention studies.
Docosahexaenoic acid (DHA), beta-glucan (BG) and anthocyanins (AC) extracts and enriched ingredients will be extracted and produced at the large scale by the responsible project partners (ASL, SOF and ABRO) respectively on the basis of formulation and processes set up in WP2 activities. The protocols for the formulation of BEF within the 3 selected food matrices (bakery, dairy and egg) delivered by WP2 will be adopted and BEF will be produced with a shelf life established, taking into consideration the safety issues. In particular, DPL will produce cereal-based BEF, ADX dairy-based and egg-based BEF.
1. To produce chemically well-characterised and safe bioactives to be added to selected foods.
2. To produce safe BEF containing the concentrations of bioactives required for food functionality in subsequent WPs.
WP5 Dietary pilot and intervention studies (leader: Luigi Ricciardiello – UNIBO)
The main aim of WP5 is to perform the intervention studies which are essential to provide information on the effectiveness of bioactive-enriched foods (BEF). Within WP5, pilot studies will be performed using the best BEFs from the three different matrices, selected in WP2 and produced in WP4. This will identify which enrichment process within each food matrix is the most effective in reducing blood triglycerides (TG) and increasing HDL-cholesterol (HDL-C) levels. The most active BEF within each matrix will be then used in a larger randomized, placebo-controlled intervention study. Since pilot and intervention studies involve human beings, particular care will be devoted by PATHWAY-27 consortium to the respect of all human rights and general ethical principles.
The specific objectives of this WP will be to:
1. Evaluate, for each food matrix, the possible synergistic effect of DHA and other selected bioactives;
2. Select through pilot studies, the most effective BEF within each food matrix;
3. Describe detailed best practices for the intervention study (including SOPs for harmonizing analytical methodologies, determination of primary and secondary endpoints, power calculations, recruitment methodology, inclusion and exclusion criteria, baseline measurements, interventions, final measurements and statistical analyses);
4. Conduct larger dietary intervention studies using the best BEF (one for each matrix, identified by the pilot studies) so as to evaluate the impact of the food matrix on the effectiveness of bioactives.
WP6 Understanding the mechanisms underlying the effects of the PATHWAY-27 bioactive enriched foods (leader: Clarissa Gerhauser – DKFZ)
The main aim of WP 6 is to understand the mechanisms underlying the effects observed on primary and secondary endpoints related to the consumption of bioactive-enriched foods (BEF).
WP6 will clarify how the different food matrices may influence the effects of the bioactives. For this, digestibility of BEF and bioavailability of bioactives will be determined in vivo in pigs, and bioactive concentrations will be evaluated in blood and faeces of volunteers. WP6 will also evaluate whether single nucleotide polymorphisms (SNPs) in selected genes related to MS will influence the response to BEF.
The use of different advanced omics techniques will help clarify whether mechanisms observed or previously described in vitro have a role also in vivo. Results obtained in WP3 will be used as a guide for WP6 which, in turn, will give important inputs for WP3 activities.
Some tasks in WP6 will be performed in subgroups of volunteers and their selection will be done in double-blind using a dedicated software designed in WP5. This will allow examination of blind balanced subgroups of volunteers and the start analyses before the completion of the intervention study.
WP7 Guidelines for the substantiation of nutritional and health claims on bioactive enriched foods (leader: Stephane Vidry – ILSI)
Preparation, publication and implementation of guidance documents that will inform and assist the food industry sector, especially SMEs, to produce bioactive-enriched foods (BEF) with supportive health claims according to the EU legislation, thereby supporting implementation of the legislation claim framework and enhancing cooperation between complementary scientific disciplines, public-private partnership in the food sector, stakeholder interactions, and - ultimately - the health of consumers. Guidelines and best practice on preparation of scientifically-correct standard dossiers necessary to obtain a positive evaluation by EFSA will also consider legal and labeling aspects, as well as gender and age issues, whenever appropriate. The results of WP7 will be readily exploited and disseminated via WP8.
WP8 Dissemination, exploitation and technology transfer (leader: Andras Sebok – CBHU)
The general objective of this work package is to ensure that the results of the project will be extensively communicated, disseminated and transferred to the main stakeholders: industry, SMEs, policymakers and regulators, food safety and food control authorities, retailers, intermediaries, NGO, consumers, etc. The dissemination tasks will be performed in an easily-understandable format facilitating the practical application of project results. The main focus of this WP will be aimed at promoting and ensuring the effective transfer of foreground to the European food industry, SMEs, Policy Makers, and the general public, with the main aim of increasing the European innovation potential and competitiveness.
The WP8 specific objectives include:
• To promote widely the communication of the project's research findings in dedicated events (e.g. Conferences and workshops across Europe, final project open meeting, etc.)
• To communicate the results in a targeted manner to professional stakeholders and the general public by means of reports, brochures, and dedicated communication tools;
• To ensure and present the results of dedicated training courses and internet-based applications (e.g. knowledge portal) from RTD experts to industry technicians;
• To organize joint trainings for scientists and technologists regarding processes and protocols developed to ensure the correct combination of the bioactives demonstrated having the features for health claims into appropriate food matrices (so ensuring also their nutritional claims for uptake by the market).
To pursue these goals the consortium will dedicate special attention to the rules of the Annex II to the Grant Agreement and of the Consortium Agreement related to IPR matters and publications.