The use of adverse outcome pathways in the safety evaluation of food additives

Funder: ILSI EuropeAbstract: In the last decade, adverse outcome pathways have been introduced in the fields of toxicology and risk assessment of chemicals as pragmatic tools with broad application potential. While their use in the pharmaceutical and cosmetics sectors has been well documented, their application in the food area remains largely unexplored. In this respect, an expert group of the International Life Sciences Institute Europe has recently explored the use of adverse outcome pathways in the safety evaluation of food additives. A key activity was the organization of a workshop, gathering delegates from the regulatory, industrial and academic areas, to discuss the potentials and challenges related to the application of adverse outcome pathways in the safety assessment of food additives. The present paper describes the outcome of this workshop followed by a number of critical considerations and perspectives defined by the International Life Sciences Institute Europe expert group

Characterizing the coverage of critical effects relevant in the safety evaluation of food additives by AOPs

Abstract: There is considerable interest in adverse outcome pathways (AOPs) as a means of organizing biological and toxicological information to assist in data interpretation and method development. While several chemical sectors have shown considerable progress in applying this approach, this has not been the case in the food sector. In the present study, safety evaluation reports of food additives listed in Annex II of Regulation (EC) No 1333/2008 of the European Union were screened to qualitatively and quantitatively characterize toxicity induced in laboratory animals. The resulting database was used to identify the critical adverse effects used for risk assessment and to investigate whether food additives share common AOPs. Analysis of the database revealed that often such scrutiny of AOPs was not possible or necessary. For 69% of the food additives, the report did not document any adverse effects in studies based on which the safety evaluation was performed. For the remaining 31% of the 326 investigated food additives, critical adverse effects and related points of departure for establishing health-based guidance values could be identified. These mainly involved effects on the liver, kidney, cardiovascular system, lymphatic system, central nervous system and reproductive system. AOPs are available for many of these apical endpoints, albeit to different degrees of maturity. For other adverse outcomes pertinent to food additives, including gastrointestinal irritation and corrosion, AOPs are lacking. Efforts should focus on developing AOPs for these particular endpoints

VALUTAZIONE DEL DANNO PRIMARIO ED OSSIDATIVO AL DNA IN LINFOCITI SALIVARI PER IL MONITORAGGIO DEGLI EFFETTI BIOLOGICI PRECOCI CAUSATI DALL’INQUINAMENTO ATMOSFERICO NEI BAMBINI: STUDIO MAPEC

VALUTAZIONE DEL DANNO PRIMARIO ED OSSIDATIVO AL DNA IN LINFOCITI SALIVARI PER IL MONITORAGGIO DEGLI EFFETTI BIOLOGICI PRECOCI CAUSATI DALL’INQUINAMENTO ATMOSFERICO NEI BAMBINI: STUDIO MAPEC Samuele Vannini (1) - Sara Levorato (1) - Elisabetta Ceretti (2) - Milena Villarini (1) - Silvia Bonetta (3) - Cristina Fatigoni(1) - Annalaura Carducci (4) - Tania Salvatori (1) - Maria Rosaria Tumolo (5) - Alessio Perotti (6) - Silvia Bonizzoni (7) - Alberto Bonetti (8) - Massimo Moretti (1) - Umberto Gelatti (2) Dipartimento di Scienze Farmaceutiche, Università degli Studi di Perugia, Perugia, Italia (1) - Dipartimento di Specialità Medico-Chirurgiche, Scienze Radiologiche e Sanità Pubblica, Università degli Studi di Brescia, Brescia, Italia (2) - Dipartimento di Scienze della Sanità Pubblica e Pediatriche, Università degli Studi di Torino, Torino, Italia (3) - Dipartimento di Biologia, Università degli Studi di Pisa, Pisa, Italia (4) - Dipartimento di Scienze e Tecnologie Biologiche ed Ambientali, Università del Salento, Lecce, Italia (5) - Dipartimento di Bioscienze, Università del Studi di Parma, Parma, Italia (6) - Ufficio Minori, Comune di Brescia, Brescia, Italia (7) - Centro Servizi Multisettoriale e Tecnologico, Csmt Gestione S.c.a.r.l., Brescia, Italia (8) Il particolato è l’inquinante atmosferico che provoca i maggiori danni alla salute umana in Europa (Agenzia Europea dell’Ambiente; www.eea.europa.eu). Tra le prime 30 città più inquinate del continente, oltre la metà sono italiane, in particolare quelle situate nella pianura Padana, con concentrazioni di PM10, PM2,5 e NOx ben al di sopra dei valori di riferimento europei. Studi epidemiologici hanno rilevato un’associazione positiva tra esposizione ad inquinamento atmosferico, soprattutto al PM, ed incidenza e mortalità per diverse malattie cronico-degenerative, come cancro al polmone, malattie cardiovascolari e diabete. I bambini, in particolar modo, risultano esposti ad un elevato rischio per quanto riguarda gli effetti a breve e lungo termine dell’inquinamento atmosferico. Dati recenti suggeriscono che alcune alterazioni genetiche che si verificano nella prima infanzia posso incrementare il rischio di malattie cronico-degenerative in età adulta. L’obiettivo dello studio MAPEC (Monitoring Air Pollution Effects on Children for supporting Public Health policy) è quello di valutare l’associazione tra inquinamento atmosferico e biomarcatori di effetti genotossici precoci nei bambini, e di proporre un modello per la stima del rischio globale causato dagli inquinanti atmosferici. Lo studio ha previsto il reclutamento di circa 1.000 bambini, di età compresa tra 6-8 anni, in cinque città italiane (200 bambini per città) caratterizzate da diverse concentrazioni di inquinanti atmosferici: Brescia, Torino, Pisa, Perugia e Lecce. Campioni di saliva sono stati raccolti in due differenti stagioni, inverno e primavera, caratterizzate da differenze qualitative e quantitative degli inquinanti atmosferici (1.000 × 2 = 2.000 campioni). Nei linfociti salivari è stato valutato il danno primario e ossidativo al DNA mediante il test della microgel elettroforesi su singole cellule (comet assay). Alla conferenza verranno presentati i risultati preliminari relativi al campionamento invernale

The use of adverse outcome pathways in the safety evaluation of food additives

Funder: ILSI EuropeAbstract: In the last decade, adverse outcome pathways have been introduced in the fields of toxicology and risk assessment of chemicals as pragmatic tools with broad application potential. While their use in the pharmaceutical and cosmetics sectors has been well documented, their application in the food area remains largely unexplored. In this respect, an expert group of the International Life Sciences Institute Europe has recently explored the use of adverse outcome pathways in the safety evaluation of food additives. A key activity was the organization of a workshop, gathering delegates from the regulatory, industrial and academic areas, to discuss the potentials and challenges related to the application of adverse outcome pathways in the safety assessment of food additives. The present paper describes the outcome of this workshop followed by a number of critical considerations and perspectives defined by the International Life Sciences Institute Europe expert group

Characterizing the coverage of critical effects relevant in the safety evaluation of food additives by AOPs

Abstract: There is considerable interest in adverse outcome pathways (AOPs) as a means of organizing biological and toxicological information to assist in data interpretation and method development. While several chemical sectors have shown considerable progress in applying this approach, this has not been the case in the food sector. In the present study, safety evaluation reports of food additives listed in Annex II of Regulation (EC) No 1333/2008 of the European Union were screened to qualitatively and quantitatively characterize toxicity induced in laboratory animals. The resulting database was used to identify the critical adverse effects used for risk assessment and to investigate whether food additives share common AOPs. Analysis of the database revealed that often such scrutiny of AOPs was not possible or necessary. For 69% of the food additives, the report did not document any adverse effects in studies based on which the safety evaluation was performed. For the remaining 31% of the 326 investigated food additives, critical adverse effects and related points of departure for establishing health-based guidance values could be identified. These mainly involved effects on the liver, kidney, cardiovascular system, lymphatic system, central nervous system and reproductive system. AOPs are available for many of these apical endpoints, albeit to different degrees of maturity. For other adverse outcomes pertinent to food additives, including gastrointestinal irritation and corrosion, AOPs are lacking. Efforts should focus on developing AOPs for these particular endpoints

Lifestyles and socio-cultural factors among children aged 6-8 years from five Italian towns: The MAPEC-LIFE study cohort

Background: Lifestyles profoundly determine the quality of an individual’s health and life since his childhood. Many diseases in adulthood are avoidable if health-risk behaviors are identified and improved at an early stage of life. The aim of the present research was to characterize a cohort of children aged 6–8 years selected in order to perform an epidemiological molecular study (the MAPEC_LIFE study), investigate lifestyles of the children that could have effect on their health status, and assess possible association between lifestyles and socio-cultural factors. Methods: A questionnaire composed of 148 questions was administered in two different seasons to parents of children attending 18 primary schools in five Italian cities (Torino, Brescia, Pisa, Perugia and Lecce) to obtain information regarding the criteria for exclusion from the study, demographic, anthropometric and health information on the children, as well as some aspects on their lifestyles and parental characteristics. The results were analyzed in order to assess the frequency of specific conditions among the different seasons and cities and the association between lifestyles and socio-economic factors. Results: The final cohort was composed of 1,164 children (50.9 boys, 95.4% born in Italy). Frequency of some factors appeared different in terms of the survey season (physical activity in the open air, the ways of cooking certain foods) and among the various cities (parents’ level of education and rate of employment, sport, traffic near the home, type of heating, exposure to passive smoking, ways of cooking certain foods). Exposure to passive smoking and cooking fumes, obesity, residence in areas with heavy traffic, frequency of outdoor play and consumption of barbecued and fried foods were higher among children living in families with low educational and/or occupational level while children doing sports and consuming toasted bread were more frequent in families with high socio-economic level. Conclusions: The socio-economic level seems to affect the lifestyles of children enrolled in the study including those that could cause health effects. Many factors are linked to the geographical area and may depend on environmental, cultural and social aspects of the city of residence

Novel approaches to derive points of departure for food chemical risk assessment

Food chemical risk assessment characterizes the potential hazards and the associated health risks resulting from exposure of humans to chemicals present in food over a specified period. This requires the identification of points of departure, which are usually derived from apical toxicity endpoints in in vivo studies. However, a shift in the traditional paradigm in toxicity testing for risk assessment has started, and increasing attention is now being paid to the use of mechanistic-based and exposure-based approaches, including toxicogenomics and quantitative in vitro-to-in vivo extrapolation. This review outlines the most recent examples of application of these new approaches to derive points of departure in food chemical risk assessment, as well as the challenges limiting their full application and future directions.</p

The use of adverse outcome pathways in the safety evaluation of food additives.

In the last decade, adverse outcome pathways have been introduced in the fields of toxicology and risk assessment of chemicals as pragmatic tools with broad application potential. While their use in the pharmaceutical and cosmetics sectors has been well documented, their application in the food area remains largely unexplored. In this respect, an expert group of the International Life Sciences Institute Europe has recently explored the use of adverse outcome pathways in the safety evaluation of food additives. A key activity was the organization of a workshop, gathering delegates from the regulatory, industrial and academic areas, to discuss the potentials and challenges related to the application of adverse outcome pathways in the safety assessment of food additives. The present paper describes the outcome of this workshop followed by a number of critical considerations and perspectives defined by the International Life Sciences Institute Europe expert group

Primary and oxidative DNA damage in salivary leukocytes as a tool for the evaluation of air pollution early biological effects in children: current status of the MAPEC (Monitoring Air Pollution Effects on Children for supporting public health policy) study

Background - Air pollution is a global problem: airborne or deposited pollutants are present everywhere on the planet, from highly polluted to remote areas. Twenty per cent of the EU urban population lives in areas where the EU air quality 24-hour limit value for particulate matter (PM10) is exceeded. At present, PM is Europe's most problematic pollutant in terms of harm to health, as reported by European Environmental Agency (EEA) in the EEA Technical Report on Air quality in Europe, 2011. Among the top 30 most polluted cities in Europe, more than half are Italian; in particular, the Po Valley is the most polluted area in Europe, with the concentrations of PM10, PM2,5 and NOx being well above the EU reference values for many days of the year. Epidemiological studies have found a consistent association between exposure to air pollution, especially to PM, and the incidence and mortality for several chronic diseases such as lung cancer, cardiovascular diseases and diabetes (Lanki et al., 2015). Among the mechanisms responsible for these adverse effects, genotoxic damage is of particular concern. Children are a high risk group with respect to the short- and long-term effects of air pollution. Indeed recent data suggest that genetic damage occurring early in childhood can increase the risk of chronic diseases, including cancer, in adulthood. The aim of the MAPEC (Monitoring Air Pollution Effects on Children for supporting public health policy) study is to evaluate the associations between air pollution and biomarkers of early biological effects in children, and to propose a model for estimating the global risk of early biological effects due to air pollutants and other factors in children (Feretti et al., 2013). Methods and analysis – Enrollment of children was performed in five Italian towns characterized by different concentrations of air polluntants: Brescia and Torino, in the Po Valley, in the Northern Italy, Pisa and Perugia located in Central Italy, and Lecce, in Southern Italy. The towns in Central and Southern Italy show lower PM values than towns in Northern Italy allowing us to make valuable comparisons between areas with higher and lower air pollution. About 1,000 children (200 children per town) aged 6-8 years were recruited in the study from first grade schools. The sample size of the study was determined in order to detect statistically significant differences in the studied parameters (i.e., primary DNA damage in lymphocytes from saliva and MN in buccal mucosa cells) among children living in towns with high air pollution and those living in less polluted towns. Biological samples were collected (1,000 × 2 = 2,000 samples) at two separate times, in winter and in late spring. In order to collect the cells for the comet assay, the children were asked to rinse their mouths twice with mineral water and the mouthwashes were then collected in tubes containing 25 ml of saline solution (NaCl 0.9%) to obtain leukocytes (Osswald et al., 2003). Leukocytes were processed in order to evaluate primary and oxidative (FPG) DNA damage caused by exposure to air pollutants. The comet assay in alkaline conditions (pH>13) was performed according to the standard protocol as well as to detect oxidative damage using FPG incubation (Azqueta et al., 2013). A medium-throughput approach (Shaposhnikov et al., 2010) was applied to the comet assay by using 12-gel units (Severn Biotech Ltd., UK). The slides (coded) were sent to the University of Parma for microscope analysis (blind). Results - The 12-gel slides were stained with ethidium bromide and examined under a fluorescence microscope; 50 randomly selected comets per spot (two spots/subject) were analysed. DNA damage was measured using the tail intensity (% DNA) parameter (Comet Assay IV, Perceptive Instruments, UK. Microscope analysis of cells sampled on winter 2014 is still ongoing. Preliminary data will be presented at the conference. Conclusions - The main objective of the MAPEC study is to evaluate the associations in children between air pollutants and early biological effects, and to propose a model for estimating the global genotoxic risk

Characterizing the coverage of critical effects relevant in the safety evaluation of food additives by AOPs.

There is considerable interest in adverse outcome pathways (AOPs) as a means of organizing biological and toxicological information to assist in data interpretation and method development. While several chemical sectors have shown considerable progress in applying this approach, this has not been the case in the food sector. In the present study, safety evaluation reports of food additives listed in Annex II of Regulation (EC) No 1333/2008 of the European Union were screened to qualitatively and quantitatively characterize toxicity induced in laboratory animals. The resulting database was used to identify the critical adverse effects used for risk assessment and to investigate whether food additives share common AOPs. Analysis of the database revealed that often such scrutiny of AOPs was not possible or necessary. For 69% of the food additives, the report did not document any adverse effects in studies based on which the safety evaluation was performed. For the remaining 31% of the 326 investigated food additives, critical adverse effects and related points of departure for establishing health-based guidance values could be identified. These mainly involved effects on the liver, kidney, cardiovascular system, lymphatic system, central nervous system and reproductive system. AOPs are available for many of these apical endpoints, albeit to different degrees of maturity. For other adverse outcomes pertinent to food additives, including gastrointestinal irritation and corrosion, AOPs are lacking. Efforts should focus on developing AOPs for these particular endpoints