Immunoproteomics of food allergens, modified allergen derivates and allergenic digestion resistant peptides.

Alergeni hrane predstavljaju jedan od uzroka anafilaksije u svetu, a samim tim i javni zdravstveni problem. U cilju razvoja efikasnije i bezbednije imunoterapije pacijenata alegičnih na hranu, neophodno je da se radi na identifikaciji i karakterizaciji uzročnika. U molekulskoj alergologiji imunoproteomika je pronašla savršenu primenu kako za identifikaciju, tako i za bolje razumevanje alergena, što tradicionalne tehnike nekada nisu bile u mogućnosti.U ovoj tezi imunoproteomikom su ispitivani alergeni hrane. Kod novog sindroma alergije na hranu, pokazali smo da IgE antitela iz seruma pacijenta alergičnih na crveno meso prepoznaju više proteina iz različito termički obrađenog goveđeg mesa, od kojih su mnogi proteini prepoznati od strane anti-α-Gal antitela. MS/MS analiza proteinskih tačaka koje su vezivale IgE rezultiralo je u identifikaciji 18 proteina. Od ovih 18 proteina identifikovano je sedam novih IgE-vezujućih proteina koji sadrže α-Gal epitop, od kojih je četiri stabilno na termičku obradu. Samim tim, alergenost proteina iz crvenog mesa je očuvana čak i nakon različite termičke obrade.Ovim je pokazano da je α-Gal epitop uobičajeno prisutan u IgE-reaktivnim proteinima iz goveđeg mesa, prepoznatih od strane seruma pacijenata alergičnih na crveno meso.Alergeni kikirikija predstavljaju najznačajnije uzročnike alergije na hranu, od kojih su konglutini, Ara h 2 i Ara h 6 najrelevantniji. Ovi proteini sadrže visoko-strukturirano jezgro, otporno na digestiju i visoku temperaturu, koje se drži zahvaljujući nekoliko disulfidnih mostova. U cilju razvoja imunoterapije za tretman alergije na kikiriki optimizovana je metoda za proizvodnju alergoida. Redukovanjem disulfidnih mostova, zatim alkilovanjem, pokazali smo da su sve disulfidne veze prekinute. Kao rezultat toga došlo je do promene u sekundarnoj i trecijarnoj strukturi proteina, kao i do gotovo gubitka IgE-vezivanja. Međutim, korišćenjem limitiranih uslova modifikacije dolazi do stvaranja intermedijernih produkata u kojima dolazi do izmene disulfida, i koji su...Food allergens are the one of the cause for development of anaphylaxis in the world, and therefore the public health problem. For better management of patients it is necessary to work on the identification and characterization of causative agents in order to find an approach that would help in the development of immunotherapy. Immunporoteomics in molecular allergology found the perfect application for identifying and better understanding of allergens, which traditional techniques sometimes were not able to.In this thesis we examined food allergens by immunoproteomics. In the new syndrome of food allergy, we have shown that IgE antibodies from the sera of the patients allergic to red meat recognize multiple proteins from different thermally processed beef, many of which have been recognized by anti-α-Gal antibodies. MS/MS analysis of protein spots, which were able to bind IgE, resulted in the identification of 18 proteins. Seven novel α-Gal-containing IgE-binding proteins were identified, of which four were stable to heat treatment. Thus, the allergenicity of red meat proteins is preserved even upon different thermal cooking. By this study we demonstrated that the α-Gal epitope is commonly present in IgE-reactive beef proteins recognized by meat-allergic patients.Peanut allergens are the most important causes of food allergies, where conglutins, Ara h 2 and Ara h 6 are the most relevant allergens. These proteins contain a highly-structured protein core, resistant to digestion and a high temperature, which is held together by several disulfide bridges. In order to develop immunotherapy for peanut allergy we optimized the method for production of allergoids. By conglutin treatment with reduction agent, and subsequently with alkylation agent, we have shown that all disulfide bonds were reduced. Disulfide bond reduction resulted in a change in the protein secondary and tertiary structure, as well as in the loss of IgE-binding. However,..

GOAL ORIENTATION PRESCHOOL PROGRAMS (CURRICULUMS) IN THE REPUBLIC OF SLOVENIA REPUBLIC OF CROATIA AND REPUBLIC OF SERBIA

U radu analiziramo i kompariramo strukturu i sadržaj aktuelnih predškolskih programa (kurikuluma) u R Sloveniji, R Hrvatskoj i R Srbiji kroz detaljan prikaz ciljnih orijentacija u njima. Primenom deskriptivne metode i postupaka analize sadržaja dokumenata, aktuelnih predškolskih programa (kurikuluma) u R Sloveniji, R Hrvatskoj i R Srbiji ciljne orijentacije ovih programskih koncepcija smo pokušali sagledati kroz utvrđena polazišta za njihovu izradu i tumačenje prirode deteta. Rezultati našeg istraživanja ukazuju na različit sadržaj i strukturu ovih predškolskih programa (kurikuluma), i na slične programske koncepcije odnosno na istu humanističko vrednosnu orijentaciju predškolskih programa (kurikuluma), tj.identične ciljne orijentacije programa.This paper analyzes and comare the structure and content of current preschool programs (curriculums) in the Republic of Slovenia, Republic Croatia and Rebublic of Serbia with a detailed review of goal orientation in them. By using descriptive method and content analysis procedure documents, analyzes of structure of current preschool programs (curriculums ) in the of Republic of Slovenia, Republic of Croatia and Republic of Serbia, goal orientation of the program concepts we have tried to examine the established starting point for their design and interpretation of the nature of the child. The results of our survey show that there is a different content and structure of these preschool programs (curriculums), and similar program concepts, that is, to the same humanistic-valued orientation of preschool programs that identical goal orientation program

L\u27ANALISI COMPARATIVA DEGLI SCOPI E COMPITI DELL\u27 EDUCAZIONE PRESCOLASTICA E DELL\u27 ISTRUZIONE SECONDO I CONCETTI PROGRAMMATICI IN SLOVENIA, CROAZIA E SERBIA

U radu su predstavljeni programi (kurikulumi) predškolskog odgoja i obrazovanja u Sloveniji, Hrvatskoj i Srbiji kroz detaljni prikaz ciljeva i zadataka predškolskog odgoja u ovim društvima. Primjenjena je deskriptivna metoda i postupak raščlanjivanja sadržaja aktuelnih programskih dokumenata. Polazeći od činjenice da se programi (kurikulumi), po sadržaju i strukturi međusobno razlikuju, rezultati naših teorijskih razmatranja ukazuju na iste temeljne ciljeve institucionalnog predškolskog odgoja i obrazovanja u ovim društvima. Javljaju se i razlike u konkretizaciji općih, globalnih i pojedinačnih ciljeva u ovim programima (kurikulumima), kojima se potiče razvitak djeteta, čime potvrđujemo u predškolskim programima (kurikulumima) u Sloveniji, Hrvatskoj i Srbiji utvrđenost različito postavljenih odgojno-obrazovnih ciljeva, u odnosu na dijete i njegove aspekte razvitka.U radu su predstavljeni programi (kurikulumi) predškolskog odgoja i obrazovanja u Sloveniji, Hrvatskoj i Srbiji kroz detaljni prikaz ciljeva i zadataka predškolskog odgoja u ovim društvima. Primjenjena je deskriptivna metoda i postupak raščlanjivanja sadržaja aktualnih programskih dokumenata. Polazeći od činjenice da se programi (kurikulumi), po sadržaju i strukturi međusobno razlikuju, rezultati naših teorijskih razmatranja ukazuju na iste temeljne ciljeve institucionalnog predškolskog odgoja i obrazovanja u ovim društvima. Javljaju se i razlike u konkretizaciji općih, globalnih i pojedinačnih ciljeva u ovim programima (kurikulumima) kojima se potiče razvitak djeteta, čime potvrđujemo u predškolskim programima (kurikulumima) u Sloveniji, Hrvatskoj i Srbiji utvrđenost različito postavljenih odgojno-obrazovnih ciljeva u odnosu na dijete i njegove aspekte razvitka.This paper presents programs (curricula) of preschool upbringing and education in Slovenia, Croatia and Serbia through detailed overview of aims and goals of preschool education in these societies. Descriptive method and the method of content analysis of current program documents were applied. Based on the fact that the programs (curricula) are different in content and structure, the results of our theoretical considerations point to the same basic goals of institutional preschool upbringing and education in these societies. There are also differences in the concretization of general, global and individual goals in these programs (curricula) that encourage child\u27s development, confirming that in Slovenia, Croatia and Serbia different educational goals in relation to the child and its development aspects are set.Nella tesi sono presentati i programmi (curricula) di educazione e formazione prescolare nella Slovenia, Croazia e Serbia, attraverso la rappresentazione dettagliata degli obiettivi e dei compiti di educazione prescolare nelle società sopraindicate. È stato applicato il metodo descrittivo e l’analisi del contenuto di attuali documenti del programma. Partendo dal fatto che i programmi (curricula) sono diversi per quanto riguarda il contenuto e la struttura, il risultato delle nostre riflessioni teoriche puntano sugli stessi obiettivi di base della formazione ed educazione istituzionale prescolare nelle società menzionate. Troviamo le differenze nel processo di concretizzazione di obiettivi generali, globali e quelli particolari in questi programmi (curricula) che stimolano lo sviluppo del bambino, confermando così che gli nei programmi (curricula) prescolari nella Slovenia, Croazia e Serbia, obiettivi di educazione e formazione rispetto al bambino e agli aspetti del suo sviluppo sono posti in modo diverso

Supplementary data for article: Stanić-Vučinić, D.; Prodić, I.; Apostolović, D.; Nikolić, M.; Ćirković-Veličković, T. Structure and Antioxidant Activity of Beta-Lactoglobulin-Glycoconjugates Obtained by High-Intensity-Ultrasound-Induced Maillard Reaction in Aqueous Model Systems under Neutral Conditions. Food Chemistry 2013, 138 (1), 590–599. https://doi.org/10.1016/j.foodchem.2012.10.087

Supplementary material for: [https://doi.org/10.1016/j.foodchem.2012.10.087]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/1583

Enterocytes in Food Hypersensitivity Reactions

Food hypersensitivity reactions are adverse reactions to harmless dietary substances, whose causes are hidden within derangements of the complex immune machinery of humans and mammals. Until recently, enterocytes were considered as solely absorptive cells providing a physical barrier for unwanted lumen constituents. This review focuses on the enterocytes, which are the hub for innate and adaptive immune reactions. Furthermore, the ambiguous nature of enterocytes is also reflected in the fact that enterocytes can be considered as antigen-presenting cells since they constitutively express major histocompatibility complex (MHC) class II molecules. Taken together, it becomes clear that enterocytes have an immense role in maintaining oral tolerance to foreign antigens. In general, the immune system and its mechanisms underlying food hypersensitivity are still unknown and the involvement of components belonging to other anatomical systems, such as enterocytes, in these mechanisms make their elucidation even more difficult. The findings from studies with animal models provide us with valuable information about allergic mechanisms in the animal world, while on the other hand, these models are used to extrapolate results to the pathological conditions occurring in humans. There is a constant need for studies that deal with this topic and can overcome the glitches related to ethics in working with animals

Post-translational modifications (PTMs) of major peanut allergen epitopes occurring as a result of purification process

Peanut allergy affects a large portion of world population causing reactions ranging from mild to severe. Seed storage proteins Ara h 1, Ara h 2, Ara h 3 and Ara h 6 are peanut allergens, with well characterized IgE epitopes but little is known about PTMs effect on their properties. Our aim was to investigate PTMs present on known epitopes of said purified proteins using bottom-up proteomcs methods. Out of the 4 allergens, Ara h 2 has the highest number of PTMs (14), while the most common are hydroxylation Pro, dehydration and deamidation (N, Q). Peanut allergen epitopes are indeed carriers of PTMs that could affect protein allergenicity and digestibility. Further investigation on peanut extracts prepared in different ways is necessary in order to fully understand the impact protein modifications could have on their allergenic potential

Supplementary data for article: Vesic, J.; Stambolic, I.; Apostolovic, D.; Milcic, M.; Stanic-Vucinic, D.; Cirkovic Velickovic, T. Complexes of Green Tea Polyphenol, Epigalocatechin-3-Gallate, and 2S Albumins of Peanut. Food Chemistry 2015, 185, 309–317. https://doi.org/10.1016/j.foodchem.2015.04.001

Supplementary material for: [https://doi.org/10.1016/j.foodchem.2015.04.001]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/1718

Major peanut allergen Ara h1 and Ara h 3 epitope post-translational modifications (PTMs)

Objective: Peanuts are widely used for the preparation of a variety of foods and are also relied on as a protein extender. Peanut allergies affect a large portion of world population causing reactions ranging from mild to severe that can lead to anaphylaxis and even death. Seed storage proteins Ara h 1 and Ara h 3 are known as major peanut allergens. IgE epitopes of these allergens have been characterized, but little is known about how post-translational modifications (PTMs) affect their allergenicity and digestibility. Our aim was to investigate PTMs present on known epitopes of said proteins using bottom-up proteomcs methods. Material and Methods: Purified 2S albumins (Ara h 1 and Ara h 3) were analysed by a Top5 nLC- MS/MS method by LTQ Orbitrap XL (Thermo Fischer Scientific, Germany). Spectra were compared to Uniprot derived Peanut protein database, hybridized with the Repository of Adventitious Proteins (cRAP), using Peaks 8.5 software package (BSI, Canada). Epitopes were searched for possible PTMs by matching PEAKS PTM results with mapped positions of epitope sequences (found in the Immune Epitope Database – IEDB www.iedb.org). Results: According to IEDB Ara h 1 contains 327 peptide epitopes, within which we detected 8 likely PTMs. Hydroxylation Pro and pyro-glu from Q were found as most common in Ara h 1 epitopes. Ara h 3 has only 110 epitopes, according to IEDB with 10 likely PTMs. Hydroxylation Pro, dehydration and methylation (KR) were found as most frequent in Ara h 3 epitopes. PTMs could be found in the vicinity of trypsin cleavage sites, which could have an impact on digestibility. Conclusions: Peanut allergen epitopes are indeed carriers of PTMs. These results show promise in revealing a possible role PTMs could have on protein allergenicity and digestibility. Further investigation is necessary in order to fully understand the impact protein modifications could have on their allergenic potentialBook of Abstract