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Background: The introduction of whole new foods in a population may lead to sensitization and food allergy. This constitutes a potential public health problem and a challenge to risk assessors and managers as the existing understanding of the pathophysiological processes and the currently available biological tools for prediction of the risk for food allergy development and the severity of the reaction are not sufficient. There is a substantial body of in vivo and in vitro data describing molecular and cellular events potentially involved in food sensitization. However, these events have not been organized in a sequence of related events that is plausible to result in sensitization, and useful to challenge current hypotheses. The aim of this manuscript was to collect and structure the current mechanistic understanding of sensitization induction to food proteins by applying the concept of adverse outcome pathway (AOP). Main body: The proposed AOP for food sensitization is based on information on molecular and cellular mechanisms and pathways evidenced to be involved in sensitization by food and food proteins and uses the AOPs for chemical skin sensitization and respiratory sensitization induction as templates. Available mechanistic data on protein respiratory sensitization were included to fill out gaps in the understanding of how proteins may affect cells, cell-cell interactions and tissue homeostasis. Analysis revealed several key events (KE) and biomarkers that may have potential use in testing and assessment of proteins for their sensitizing potential. Conclusion: The application of the AOP concept to structure mechanistic in vivo and in vitro knowledge has made it possible to identify a number of methods, each addressing a specific KE, that provide information about the food allergenic potential of new proteins. When applied in the context of an integrated strategy these methods may reduce, if not replace, current animal testing approaches. The proposed AOP will be shared at the www.aopwiki.org platform to expand the mechanistic data, improve the confidence in each of the proposed KE and key event relations (KERs), and allow for the identification of new, or refinement of established KE and KERs. © 2017 The Author(s).

Pollen is routinely monitored, but it is unknown whether pollen counts represent allergen exposure. We therefore simultaneously determined olive pollen and Ole e 1 in ambient air in C ordoba, Spain, and Evora, Portugal, using Hirst-type traps for pollen and high-volume cascade impactors for allergen. Pollen from different days released 12-fold different amounts of Ole e 1 per pollen (both locations P < 0.001). Average allergen release from pollen (pollen potency) was much higher in C ordoba (3.9 pg Ole e 1/pollen) than in Evora (0.8 pg Ole e 1/pollen, P = 0.004). Indeed, yearly olive pollen counts in C ordoba were 2.4 times higher than in Evora, but Ole e 1 concentrations were 7.6 times higher. When modeling the origin of the pollen, >40% of Ole e 1 exposure in Evora was explained by high-potency pollen originating from the south of Spain. Thus, olive pollen can vary substantially in allergen release, even though they are morphologically identical.

Introduction. Exposure to allergens is pivotal in determining sensitization and allergic symptoms in individuals. Pollen grain counts in ambient air have traditionally been assessed to estimate airborne allergen exposure. However, the exact allergen content in ambient air is unknown. HIALINE therefore monitored atmospheric concentrations of Betula, Poaceae and Olea pollen grains and matched their major allergens Bet v1, Phl p5 and Ole e1 across Europe. Monitoring the allergens themselves together with pollen in ambient air might be an improvement in allergen exposure assessment. New knowledge through the use of new experimental approaches in the field of aerobiological monitoring will enable better in the prevention and clinical management of pollinosis. In order to disseminate the knowledge of the project we present the results of first year of birch pollen grains and the matched major pollen allergen Bet v1 monitored in Parma (UNIPR), Italy with a short reference to the results obtained by the other participants and about developing models of dissemination and forecasts of pollen and allergens. Materials and methods. The pollen was sampled by a Hirst pollen trap. Allergens was collected with a CHEMVOL® high-volume cascade impactor, extracted from pollen and quantified by ELISA. Antibodies for analysis of Bet v1 are delivered by the industrial partner in this project. Quality control has been carried out for the monitoring pollen activities and allergen concentrations. Results. The project has highlighted that it is possible also to measure pollen allergen in ambient air in different European areas. The results obtained from the center of Parma and other European partners have highlighted the different allergenic powers of pollen of Betula, in different geographical areas. Moreover, daily in each area the allergenic power of pollen grains was very variable. Discussion. HIALINE has been a very important project to understand the mechanisms of sensitization, clinical management of pollinosis and to improve immunotherapy towards a tailored immunotherapy. The results of the project will help medical doctors, authorities and patients, to better manage the different aspects related to pollinosis.

The observation in microscopy of histological sections allows us to evaluate structural differences, in pancreatic cells, between rats with normal glucose tolerance and with glucose intolerance (pre-diabetic) situation. Nevertheless, this pre-diabetic condition implies subtle changes in islets of Langerhans structure. This and the normal variability among sampled cells makes difficult the task of identifying glucose intolerance (pre-diabetic situation) with a low level of error. This paper presents preliminary results in the processing of histological pancreas images with the goal of identifying pre-diabetic situation in Wistar rats. The immediate goal of this work is to evaluate the performance of a classifier based in a morphometric measurement of the histological images and to assess the potential for image based automatic processing and classification. A set of 90 images, were used (58 from rats with normal glucose tolerance, and 32 from pre-diabetic ones). These images were segmented manually using ImageJ. This segmentation and area measurements have been speedup by the application of ImageJ macros which were defined for this purpose. The ratio, between the area of -cells and the islets of Langerhans , was used has the indicator of the prediabetic situation. Considering this feature, a receiver operating characteristic analysis has been performed. True positive rate, vs. false positive rate shows the predicted performance of a binary classifier as its discrimination threshold is varied.

Exposure to allergens is pivotal in determining sensitization and allergic symptoms in individuals. Pollen grain counts in ambient air have traditionally been assessed to estimate airborne allergen exposure. However, the exact allergen content of ambient air is unknown. We therefore monitored atmospheric concentrations of birch pollen grain and the matched major birch pollen allergen Bet v 1 simultaneously across Europe within the EU-funded project HIALINE (Health Impacts of Airborne Allergen\ud Information Network). Pollen count was assessed with Hirst type pollen traps at 10 l/min at sites in France,\ud United Kingdom, Germany, Italy and Finland. Allergen concentrations in ambient air were sampled at 800l/min with a Chemvol high-volume cascade impactor equipped with stages PM>10μm, 10 μm>PM>2.5μm, and in Germany also 2.5\ud μm>PM>0.12μm. The major birch pollen allergen Bet v 1 was determined with an allergen specific ELISA. Bet v 1 isoform patterns were analyzed by 2D-SDS-PAGE blots and mass spectrometric identification. Basophil activation was tested in an FcεR1-humanized rat basophil cell line passively sensitized with serum of a birch pollen lmptomatic patient. Compared to 10 previous years, 2009 was a representative birch pollen season for all stations. About 90% of the allergen was found in the PM>10μm fraction at all stations. Bet v 1 isoforms pattern did not varied substantially neither during ripening of pollen nor between different geographical locations. The average European\ud allergen release from birch pollen was 3.2 pg Bet v 1/pollen and did not vary much between the European countries. However, in all countries a >10-fold difference in daily allergen release per pollen was measured which could be explained by long range transport of pollen with a deviating allergen release. Basophil activation by\ud ambient air extracts correlated better with airborne allergen than with pollen concentration. Although Bet v 1 is a mixture of different isoforms, its fingerprint is constant across Europe. Bet v 1 was also exclusively linked to pollen. Pollen from different days varied >10-fold in allergen release. Thus exposure to allergen is inaccurately\ud monitored by only monitoring birch pollen grains. Indeed, a humanized basophil activation test correlated much better with allergen concentrations in ambient air than\ud with pollen count. Monitoring the allergens themselves together with pollen in ambient air might be an improvement in allergen exposure assessment.