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Background: Clinical and experimental analyses indicate a pathognomonic role for allergen IgE crosslinking through epitope–paratope interactions as a major initial step in the cascade leading to effector cell activation and clinical manifestations of lgE-mediated food allergies. We aimed to undertake the initial development and assessment of Ara h 2-specific IgE epitope-like peptides that can bind to allergenspecific IgE paratopes and suppress effector cell activation. Methods: We performed biopanning, screening, IgE binding, selection and mapping of peptides. We generated synthetic peptides for use in all functional experiments. ImmunoCAP inhibition, basophil and mast cell activation tests, with LAD2 cells, a human mast cell line were performed. Twenty-six children or young adults who had peanut allergy were studied. Results: We identified and selected three linear peptides (DHPRFNRDNDVA, DHPRYGP and DHPRFST), and immunoblot analyses revealed binding to lgE from peanut-allergic individuals. The peptide sequences were aligned to the disordered region corresponding to the loop between helices 2 and 3 of Ara h 2, and conformational mapping showed that the peptides match the surface of Ara h 2 and h 6 but not other peanut allergens. In ImmunoCAP inhibition experiments, the peptides significantly inhibit the binding of IgE to Ara h 2 (p < .001). In basophil and mast cell activation tests, the peptides significantly suppressed Ara h 2-induced effector cell activation (p < .05) and increased the half-maximal Ara h 2 effective concentration (p < .05). Binding of the peptides to specific IgEs did not induce activation of basophils or mast cells. Conclusions: These studies show that the indicated peptides reduce the allergenic activity of Ara h 2 and suppress lgE-dependent basophil and mast cell activation. These observations may suggest a novel therapeutic strategy for food allergy based on epitope–paratop blocking.

Alergija je bolezensko stanje, ki se pojavi kot posledica neustrezno uravnavanega imunskega odziva. Povezana je z motnjami imunske tolerance, kjer neškodljivi antigeni postanejo alergeni. Alergeni so odgovorni za sprožitev T celičnega imunskega odziva tipa 2. Alergijski imunski odziv je škodljiv za naš organizem in lahko sproži življenjsko ogrožajoče sistemske reakcije. Specifična imunoterapija je edini pristop zdravljenja vzroka alergijske bolezni in ne le njenih simptomov. Z uporabo naravnih ali rekombinatnih alergenov v specifični imunoterapiji induciramo imunsko toleranco na alergene. Toda uporaba celotnih alergenov lahko predstavlja nevarnost sprožitve anafilaktičnega šoka. Zaradi tega so v zadnjih desetletjih raziskave usmerjene v iskanje molekul z zmanjšano ali popolnoma odstranjeno alergogenostjo. Raziskave potekajo na spremenjenih alergenih, delih alergenov, peptidnih fragmentih alergenov in mimotopih alergenov. Prvi del doktorske disertacije obsega pregled alergenskih mimotopov, ki so bili odkriti z uporabo tehnologije bakteriofagnega prikaza. Pregled se nanaša predvsem na uporabnost bakteriofagnih peptidnih knjižnic. S pomočjo teh knjižnic lahko odkrijemo mimotope s potencialom za razvoj imunoterapije. Knjižnice nam pomagajo pri identifikaciji prevladujočih B celičnih epitopov in določanju IgE profilov posameznih pacientov. Odkritje epitopov in individualnih IgE profilov nam služi kot pomoč pri razvoju diagnostike in imunoterapije tako za splošno uporabo kot tudi za individualizirano zdravljenje. Z bakteriofagnimi peptidnimi knjižnicami si pomagamo tudi pri preučevanju navzkrižne reaktivnosti med alergeni. Vsi omenjeni prispevki bakteriofagnih peptidnih knjižnic so nepogrešljivi sestavni deli za razumevanje alergijske patogeneze ter za razvoj novih imunoterapij. Poleg uporabe v bakteriofagnih peptidnih knjižnicah se bakteriofagi vedno bolj uporabljajo in testirajo kot imunogeni nosilci različnih peptidov. Bakteriofagi tako združujejo odkrivanje mimotopov in imunizacijo v eno skupno platformo. V drugem delu doktorske disertacije sem se osredotočil na alergijo za mačke, ki pri alergičnemu posamezniku pogosto vodi v razvoj astme. Za zdravljenje alergije za mačke se trenutno uporabljajo le pripravki s celotnimi alergeni, kar lahko pri prevelikem odmerku vodi do sprožitve anafilaksije. Zaradi tega predstavlja identifikacija epitopov in mimotopov mačjih alergenov pomembno odkritje za razvoj učinkovite in varne specifične imunoterapije. Z uporabo tehnologije bakteriofagnega prikaza smo identificirali pet strukturnih mimotopov glavnega mačjega alergena Fel d 1. Identificirane mimotope smo računalniško prilegali na površino alergena in definirali možno epitopno mesto na samem alergenu. Z vezavo specifičnih protiteles razreda IgE iz serumov pacientov alergičnih za mačke smo potrdili IgE reaktivnost mimotopov. Mimotope smo v večjem številu izrazili na površini nitastih bakteriofagov. Bakteriofage smo izbrali kot imunogeni nosilec, ki je zmožen spodbuditi ustrezen imunski odziv. Dokazali smo, da sami mimotopi niso alergogeni, saj niso aktivirali bazofilcev, kar kaže na njihov potencial za razvoj hipoalergenih terapevtskih produktov. Bakteriofagi z izraženimi mimotopi so spodbudili močan T celični imunski odziv usmerjen v tip 1. Sami mimotopi so prispevali k tipu 1 T celičnega odziva, saj so povečali produkcijo interlevkina 2. Identificirani mimotopi in izbrani nosilec lahko pripomorejo k razumevanju in načrtovanju razvoja hipoalergenih pripravkov za zdravljenje alergije za mačke. Tema tretjega dela doktorske disertacije je bila alergija za arašide, ki predstavlja veliko nevarnost za sprožitev anafilaksije. Anafilaksija je pri alergijski reakciji posledica sistemskih odzivov in predstavlja enega glavnih razlogov, da so raziskave usmerjenje v iskanje varnih in učinkovitih produktov za peroralno imunoterapijo. S tem razlogom smo se odločili identificirati mimotope z znižano alergogenostjo v primerjavi s celotnimi alergeni. Z uporabo tehnologije bakteriofagnega prikaza smo uspešno identificirali mimotope Ara h 2, ki je glavni alergen v arašidih. Odkriti mimotopi so se prilegali na površino strukture Ara h 2, kjer se nahaja tudi dominantni epitop. Združene s sidrno domeno smo mimotope izrazili na površini Lactoccoccus lactis, katere smo uporabili kot imunogene nosilce. Tako pripravljeni konstrukti so vezali protitelesa razreda IgE iz serumov pacientov alergičnih za arašide. Identificirani mimotopi so imeli znižano alergogenost z izjemo mimotopa L7-N40, ki je ohranil delno alergogenost. Omenjeni mimotop je izkazal največjo podobnost z dominantnim epitopom. Naši konstrukti, izrazito konstrukt z mimotopom L7-N40, so bili zmožni sprožiti T celični imunski odziv. V primerjavi s celotnim alergenom so signifikantno povišali produkcijo IFN-γ, kar kaže na usmeritev imunskega odziva v tip 1 T celični odziv. Naši konstrukti z mimotopi predstavljajo zanimiv dostavni sistem z imunomodulatornimi lastnostmi za ciljanje mukoznih površin v gastrointestinalnem traktu. Doktorska disertacija je pomoč k razumevanju in razvoju varne in učinkovite specifične imunoterapije, ki bo pripomogla k zdravljenju alergije za mačke in arašide z uporabo hipoalergenih terapevtskih pripravkov. Allergy is a disease state occuring as an effect of unsuitable regulated immune response. It is connected with dysfunction of immune tolerance where harmless antigens become allergens. Allergens are responsible for eliciting type 2 T cell immune response. Allergic immune response is harmful to our organism and can trigger life-threatening systemic reactions. Specific immunotherapy is the only way to cure allergic disease. The use of natural and recombinant allergens in specific immunotherapy induces immune tolerance to allergens but represents a danger for triggering of anaphylactic shock. Hence, investigations in the last decades have been directed to finding substances with lower or completely removed allergenic activity. Research is underway on altered allergens, parts of allergens, peptides derived from allergens, and mimotopes of allergens. The first part of doctoral thesis includes review of allergen mimotopes discovered by phage display technology. Review mostly refers to usefulness of phage display peptide libraries. With the help of these libraries we can discover mimotopes with potential to develop immunotherapy. The libraries help us to identify dominant B-cell epitopes and to determine IgE profiles of individual patients. Epitope discovery and individual IgE profiles serve as help to develop diagnostics and immunotherapy for common use as for individual treatment. Phage display peptide libraries also help us in studying of cross-reactivity among allergens. All these different contributions of phage display peptide libraries are indispensable components for understanding allergic pathogenesis and for development of new immunotherapies. Beside function of bacteriophages in peptide libraries, they are increasingly used and tested as immunogenic carriers of different peptides. Bacteriophages combine mimotope discovery and immunization into one complete platform. In the second part of doctoral thesis I focused on cat allergy which often leads to asthma development in allergic individual. At present used preparations for treatment of cat allergy contain whole allergens which can lead to triggering anaphylaxis as effect of an overdose. Epitope and mimotope identification of cat allergens represents important discovery for development of effective and safe specific immunotherapy. We identified five conformational mimotopes of the major cat allergen Fel d 1 by using phage display technology. Identified mimotopes were mapped to the surface of the allergen and potential epitope spot was defined. Binding of specific IgE from sera of cat-allergic patients confirmed IgE reactivity of mimotopes. Mimotopes were displayed in multiple copies on the surface of filamentous bacteriophages which were chosen as immunogenic carriers able to elicit suitable immune response. We demonstrated that mimotopes alone are not allergenic because they did not activate basophils. This shows their potential for development of hypoallergenic therapeutical products. Prepared construct elicited strong type 1 directed T cell immune response. Mimotopes alone contributed to type 1 T cell response by increasing production of interleukin 2. Identified mimotopes and selected carriers can contribute to progress in development of hypoallergenic preparations for treatment of cat allergy. Subject of the third part of doctoral thesis was peanut allergy which represents high risk of triggering anaphylaxis in allergic individuals. Anaphylaxis, a systemic respons in allergic reaction, is a reason researches are directed into discovery of safe and effective products for oral immunotherapy. This was the reason we decided to investigate mimotopes which have lower allergenic activity in comparison with whole allergens. We successfully identified mimotopes of Ara h 2, the major peanut allergen, by using phage display technology. Discovered mimotopes were mapped to the dominant surface epitope of Ara h 2. Fused with anchoring domain we displayed mimotopes on the surface of Lactococcus lactis used as immunogenic carriers. Prepared constructs bound IgE antibodies from sera of peanut-allergic patients. Mimotopes had lower allergenic activity except for mimotope L7-N40, which partially preserved allergenic activity. Mentioned mimotope showed the highest similarity with dominant epitope. Our constructs, especially construct with mimotope L7-N40, were able to elicit T cell immune response. In comparison with allergen they had significantly higher production of IFN-γ demonstrating type 1 directed T cell response. Our constructs with mimotopes present interesting delivery system with immunomodulatory characteristics to target mucosal surfaces in gastrointestinal tract. Doctoral thesis is helping to understand and develop safe and effective specific immunotherapy which will contribute to treatment of cat and peanut allergy by using hypoallergenic therapeutical preparations.

Uvod : Astma je kronična upala dišnih putova. Uzroci su alergije, virusne respiratorne infekcije i nadražujuće tvari iz zraka. Broj oboljelih povećava se, a pritom prednjače djeca. Ishrana majke tijekom trudnoće te ishrana djeteta u prvoj godini života imaju važan utjecaj na razvoj i tijek bolesti. Metode : Upotrijebljena je narativna analiza, pregled stručne i znanstvene literature. Pretraživanje literature temeljilo se na istraživačkom pitanju Kako ishrana u ranom djetinjstvu utječe na rizik za razvoj astme i alergijskih bolesti?. Rezultati: Analizom stručne literature utvrđeno je da ishrana majke ima važnu ulogu pri razvoju astme i alergijskih bolesti kod djece. Dokazi upućuju na to da je pravilno uravnotežena i raznolika ishrana važna za djetetovo zdravlje te da smanjuje rizik od nastajanja bolesti. Stručnjaci tijekom trudnoće ne preporučuju izbjegavanje određenih namirnica kao što su kikiriki, orasi i školjke, osim u slučaju ako je majka alergična na njih. Rasprava i zaključak: Zbog povećane učestalosti astme i alergijskih bolesti, osobito kod djece, potrebno je istaknuti važnost i utjecaj raznolike ishrane tijekom trudnoće te dojenja tijekom prvih šest mjeseci djetetova života. Različite dijete tijekom i nakon trudnoće te tijekom dojenja nisu se pokazale učinkovitima u sprječavanju razvoja astme i alergijskih bolesti kod djece.

Asthma is a severe and chronic disabling disease affecting more than 300 million people worldwide. Although in the past few drugs for the treatment of asthma were available, new treatment options are currently emerging, which appear to be highly effective in certain subgroups of patients. Accordingly, there is a need for biomarkers that allow selection of patients for refined and personalized treatment strategies. Recently, serological chip tests based on microarrayed allergen molecules and peptides derived from the most common rhinovirus strains have been developed, which may discriminate 2 of the most common forms of asthma, that is, allergen- and virus-triggered asthma. In this perspective, we argue that classification of patients with asthma according to these common trigger factors may open new possibilities for personalized management of asthma., La lista completa de autores que integran el documento puede consultarse en el archivo., Facultad de Ciencias Exactas

Alergija ili preosjetljivost je pretjerana reakcija organizma izazvana tvarima koji dovode do oštećenja organizma. Reakcije preosjetljivosti u dentalnoj medicini su rijetke i najčešće se pojavljuju kontaktne ili odgođene reakcije preosjetljivosti (tip IV) na različite metale, akrilate ili ostale komponente dentalnih materijala u obliku lezija vidljivih na sluznici usne šupljine ili koži, a mogu se manifestirati i u sustavnom obliku. Legure korištene u dentalnoj medicini uslijed interakcija u usnoj šupljini podliježu elektrokemijskoj koroziji i tribokorozijskom trošenju, a brojni gradivni i pomoćni materijali klasificirani su alergeni s velikom mogućnošću izazivanja alergijskih reakcija. Korištenjem biokompatibilnih materijala čija je sigurnost potvrđena laboratorijskim i kliničkim istraživanjima želi se smanjiti mogućnost nastanka neželjenih reakcija. Najčešća metoda dokazivanja preosjetljivosti na neki materijal jest epikutano ili patch testiranje, a ukoliko pacijent u svojoj anamnezi navodi alergiju na metale ili neke druge materijale ili je preosjetljivost dokazana epikutanim testiranjem, navedeni alergogeni materijal je potrebno zamijeniti nekim manje potentnim. Allergy or hypersensitivity is an excessive reaction of the organism caused by substances that cause damage to the organism. Hypersensitivity reactions in dental medicine are rare and most often are contact or delayed hypersensitivity reactions (type IV) to various metals, acrylates or other components of dental materials in the form of lesions visible on the oral mucosa or skin, and may also manifest in systemic form. Alloys used in are subject to electrochemical corrosion and tribocorrosion wear dentistry due to interactions in the oral cavity, and a number of materials are classified as allergens with a high potential to cause allergic reactions. By using biocompatible materials whose safety has been confirmed by laboratory and clinical research, we want to reduce the possibility of adverse reactions. The most common method of proving hypersensitivity to a material is epicutaneous or patch testing, and if the patient has a history of allergy to metals or other materials, or hypersensitivity is proven by epicutaneous testing, the allergenic material should be replaced with a less potent one.

Ragweed is a prominent cause of seasonal allergies. Thus far, information on IgE-binding sites of major allergen in ragweed pollen, Amb a 1, is very limited. A powerful experimental method to gain insights on the allergen epitopes is the selection of peptides from biological libraries that bind to anti-allergen antibodies. In this work, we aimed to map IgE epitopes of Amb a 1 using epitope-mimicking short peptides % mimotopes that were affinity-selected from phage-displayed random peptide libraries. The peptides weakly aligned with the Amb a 1 primary sequence, thus suggesting that the epitopes are conformational. When the peptides were mapped onto the surface of Amb a 1 homology model, the EpiSearch analysis predicted the location of four potential epitopic sites on surface patches centred at residues K104, S110, H214, and W312. The peptides matching to the predicted epitopes bound selectively to the IgE from pool of ragweed-allergic patients% sera and therefore represent mimetics of Amb a 1 IgE epitopes. The knowledge of IgE epitopes is a prerequisite for the rational design of molecular-based approaches to diagnosis and immunotherapy of allergic diseases. Ambrozija je pomemben vzrok sezonskih alergij. Do sedaj imamo zelo malo informacij o IgE vezavnih mestih na glavnem alergenu iz cvetnega prahu ambrozije, Amb a 1. Afinitetna selekcija peptidov iz bioloških knjižnic z uporabo specifičnih protiteles proti alergenu je uporabna laboratorijska metoda za določevanje epitopov. V tej raziskavi smo s pomočjo mimotopov, kratkih peptidov, ki posnemajo epitope, izoliranih iz bakteriofagnih knjižnic naključnih peptidov, določali IgE epitope Amb a 1. Izbrani peptidi so se le šibko ujemali s primarnim zaporedjem Amb a 1, kar je nakazovalo, da so epitopi konformacijski. Da bi jih določili, smo izdelali homologni model tridimenzionalne strukture Amb a 1, na katerega smo prilegali izbrane peptide. S pomočjo programa EpiSearch smo identificirali štiri potencialne epitope na površini alergena okoli aminokislinskih ostankov K104, S110, H214 in W312. Peptidni mimetiki predvidenih epitopov so se specifično vezali na IgE iz zmesi serumov za ambrozijo alergičnih pacientov in tako predstavljajo mimetike IgE epitopov Amb a 1. Poznavanje IgE epitopov je predpogoj za racionalno načrtovanje molekularnih pristopov v diagnostiki in imunoterapiji alergijskih bolezni.

Background: Google Trends (GT) searches trends of specific queries in Google, which potentially reflect the real-life epidemiology of allergic rhinitis. We compared GT terms related to ragweed pollen allergy in American and European Union countries with a known ragweed pollen season. Our aim was to assess seasonality and the terms needed to perform the GT searches and to compare these during the spring and summer pollen seasons. Methods: We examined GT queries from January 1, 2011, to January 4, 2017. We included 15 countries with a known ragweed pollen season and used the standard 5-year GT graphs. We used the GT translation for all countries and the untranslated native terms for each country. Results: The results of “pollen,” “ragweed,” and “allergy” searches differed between countries, but “ragweed” was clearly identified in 12 of the 15 countries. There was considerable heterogeneity of findings when the GT translation was used. For Croatia, Hungary, Romania, Serbia, and Slovenia, the GT translation was inappropriate. The country patterns of “pollen,” “hay fever,” and “allergy” differed in 8 of the 11 countries with identified “ragweed” queries during the spring and the summer, indicating that the perception of tree and grass pollen allergy differs from that of ragweed pollen. Conclusions: To investigate ragweed pollen allergy using GT, the term “ragweed” as a plant is required and the translation of “ragweed” in the native language needed.

Genetski modificirani organizmi su organizmi promijenjenog genetičkog materijala koji se posljednjih tridesetak godina stvaraju tehnologijom rekombinantne DNA (genetičkim inženjerstvom). U stanicu određenog organizma unosi se gen (fragment DNA) za željenu osobinu koji se može preuzeti od druge srodne ili nesrodne vrste. Nastaju genetički preinačeni (modificirani) organizmi s novim, poboljšanim svojstvima, koji omogućuju napredak u raznim područjima ljudske djelatnosti, posebice u poljoprivredi i biotehnologiji. I dok se produkti genetički modificiranih organizama poput različitih proteina, pametnih lijekova, hormona i cjepiva prihvaćaju bez većih kritika i primjenjuju se u medicini i farmakologiji, uvođenje genetski modificirane hrane u prehranu ljudi u nekim zemljama, posebice u Europi, nailazi na velike otpore i kritike. S ciljem utvrđivanja potencijalnih rizika za zdravlje ljudi, prije izdavanja dozvole za komercijalnu proizvodnju i prodaju, svaki GM kultivar se temeljito testira i uspoređuje sa svojim konvencionalnim ekvivalentom, pa ako se utvrde razlike, rade se daljnja toksikološka i alergološka testiranja. Pritom se osobito vodi računa da se otkriju potencijalno alergeni proteini i drugi spojevi koji bi mogli djelovati toksično odnosno kancerogeno ili uzrokovati rezistentnost na antibiotike. Brojni znanstveni radovi objavljeni su na temu mogućeg rizika od konzumacije GM hrane za zdravlje ljudi. Iako jedan manji broj znanstvenih članaka ukazuje na određene rizike, prvenstveno zbog dugotrajne izloženosti GM proizvodima, ipak, ogroman broj znanstvenih radova ne nalazi dokaze o statistički značajnom riziku za zdravlje ljudi. Svjetska zdravstvena organizacija, stoga, zauzela je stav kako nema dokaza da je GM hrana opasnija za zdravlje ljudi od konvencionalne. Istovremeno više od 170 dobitnika Nobelove nagrade zalažu se za nastavak istraživanja i proizvodnju GM hrane, prvenstveno zbog brojnih koristi koje ta hrana može donijeti, kako onim zemljama koje se bore s nestašicama hrane, tako i u situacijama gdje GM hrane može unaprijediti zdravlje i opću dobrobit ljudi kao što su povećani unos nutrijenata, eliminacija postojećih alergena i toksičnih komponenti iz hrane te brojne druge prednosti., Genetically modified organisms are organisms with altered genetic material, which have been developed for nearly three decades using recombinant DNA technology (genetic engineering). Genes (fragments of DNA) containing the desired traits from either a related or unrelated species are introduced into specific organisms, thereby creating genetically changed (modified) organisms with improved features, which in turn enable significant improvements to various areas of human activity, especially in agriculture and biotechnology. While the products of genetically modified organisms, such as various proteins, smart drugs, hormones and vaccines, are generally accepted and applied in medicine and pharmacology without much criticism, in some countries, especially in Europe, the introduction of genetically modified food in nutrition has been met with great apprehension and criticism. Bearing possible health risks in mind, each genetically modified cultivar has to undergo rigorous testing, before a permit for commercial production and distribution is issued. Each genetically modified product is then also compared to its equivalent conventional counterpart, as possible differences lead to further toxicological and allergy testing. Special attention is thereby being paid to discovering potentially allergenic proteins and other compounds, which could act toxic, carcinogenic or even cause resistance to antibiotics. While numerous scientific papers pertaining to the possible health risks of consuming genetically modified food have been published, the large majority of them have not established any evidence of a statistically significant risk to human health. However, a smaller fraction of such scientific papers does highlight certain health risks with regard to the long-term exposure to genetically modified products. Consequently, The WHO has aligned itself with these findings, proclaiming that there has not been any evidence suggesting that genetically modified food is more dangerous than conventional food. Concurrently, more than 170 Nobel Prize laureates support the continuation of the research and production of genetically modified food, thereby underscoring the numerous benefits, which such food can bring about, especially in countries battling with food shortage. However, there are also situations in which genetically modified food can improve the health and general welfare of humans: an increased nutrient intake, the elimination of existing allergens and toxic compounds, as well as numerous other advantages.

Alergijske bolesti smatraju se bolestima modernog doba jer su veliki javnozdravstveni problem u industrijski razvijenim zemljama, ali i u zemljama u razvoju, posebno u djece i mlađih odraslih osoba. Uzroci porasta alergijskih bolesti još nisu dovoljno objašnjeni, no rizičnim čimbenicima smatraju se nasljedna predispozicija, izloženost alergenima, onečišćenje okoliša, smanjena stimulacija imunološkog sustava u ranim razvojnim stadijima i suvremeni način života. Alergijske bolesti treba tretirati kao općeniti poremećaj zdravlja, te se one mogu javiti u bilo koje doba života, u različitim organima istodobno ili u različitom vremenskom periodu, dugotrajne su i zahtjevaju kompleksnu medicinsku skrb. Značajno utječu na kvalitetu života, i smanjenje radne sposobnosti oboljelih te su veliko socioekonomsko opterećenje za pojedinca i društvo. U radu su teorijski obrađeni pojmovi atopijskih (alergijskih) bolesti, te pojam alergijskog rinitisa koji se smatra najčešćom manifestacijom atopije. Zatim su obrazloženi pojmovi alergije, astme i atopijskog dermatitisa. U radu su opisani načini izvođenja dijagnostičkog PRICK testa te ukazane moguće nuspojave prilikom izvođenja. Pretragom rezultata kliničkih testova ukazano je na najčešće inhalacijske alergene u sjevernom dijelu Republike Hrvatske. Objašnjena je uloga medicinske sestre u edukaciji bolesnika u atopijskim bolestima te su navedene sestrinske dijagnoze. Allergic diseases are considered to be diseases of modern time as they are a great public health problem in developed countries, but also in developing countries, especially among children and young adults. Causes of increase of allergic diseases are still not explained in detail. However, the presumption is that the risk factors are hereditary predisposition, allergen exposure, environmental pollution, decreased stimulation of immune system in early stages of development and modern lifestyle. Allergic diseases should be treated as general health problem disord, and they can appear in any period of life, in different organs at the same time or in different time period. Moreover, their duration is prolonged and they require a complex medical care. They also significantly affect the quality of life, and decrease in work ability of the ill, and they are a great socio-economic burden for the individual and the society. This paper atopic (allergic) diseases are being theoretically explained, mostly allergic rhinitis which is considered to be one of the most common manifestation of atopy. Furthermore, the terms allergy, asthma and atopic dermatitis are being explained. In the paper the way of performing the diagnostic PRICK test is pointed out as well and also the possible side effects during test performance. Statistical analysis of the skin PRICK test pointed out the most common inhalation allergens in the north part of Republic of Croatia. The role of medical nurse in education of the ill about atopic diseases is explained, and the nursing diagnoses are adduced as well.

Background: N-linked glycans present in venoms, pollen and mites are recognized by IgE antibodies from more than 20% of allergic patients but have low or no allergenic activity. Objectives: To engineer recombinant glycoproteins resembling carbohydrate-specific IgE epitopes from venoms, pollen and mites which can discriminate carbohydrate-specific IgE from allergenic, peptide-specific IgE. Methods: One or two N-glycosylation sites were engineered into the N-terminus of the non-allergenic protein horse heart myoglobin (HHM) using synthetic gene technology. HHM 1 and HHM 2 containing one or two N-glycosylation sites were expressed in baculovirus-infected High-FiveTM insect cells and a non-glycosylated version (HHM 0) was obtained by mutating the glycosylation motif. Recombinant HHM proteins were analyzed regarding fold and aggregation by circular dichroism and gel filtration, respectively. IgE reactivity was assessed by ELISA, immunoblotting and quantitative ImmunoCAP measurements. IgE inhibition assays were performed to study cross-reactivity with venom, plant and mite-derived carbohydrate IgE epitopes. Results: HHM-glycovariants were expressed and purified from insect cells as monomeric and folded proteins. The HHM-glycovariants exhibited strictly carbohydrate-specific IgE reactivity, designed to quantify carbohydrate-specific IgE and resembled IgE epitopes of pollen, venom and mite-derived carbohydrates. IgE-reactivity and inhibition experiments established a hierarchy of plant glcyoallergens (nPhl p 4>nCyn d 1>nPla a 2>nJug r 2>nCup a 1>nCry j 1) indicating a hitherto unknown heterogeneity of carbohydrate IgE epitopes in plants which were completely represented by HHM 2. Conclusion: Defined recombinant HHM-glycoproteins resembling carbohydrate-specific IgE epitopes from plants, venoms and mites were engineered which made it possible to discriminate carbohydrate- from peptide-specific IgE reactivity. Funding Statement: This study was supported by the grants P26728, F4605, F4604 and DK W1248 from the Austrian Science Fund (FWF). Rudolf Valenta is recipient of a Megagrant of the Government of the Russian Federation, grant number 14.W03.31.0024. Declaration of Interests: Rudolf Valenta has received research grants from Biomay AG, Vienna, Austria and Viravaxx, Vienna, Austria. He serves as a consultant for Biomay AG and Viravaxx. Dr. Lupinek reports personal fees from Thermo Fisher Scientific, outside the submitted work. All other authors have nothing to disclose. Ethics Approval Statement: The anonymized analysis of the sera was approved by the ethical committee of the Medical University of Vienna (EK1641/2014).

V hrani se nahajajo snovi, natančneje proteini, na katere telo pri nekaterih ljudeh reagira s preobčutljivostno reakcijo. Te so lahko blage, kot trebušni krči ali driska, lahko pa pride do življenje ogrožajočega anafilaktičnega šoka. Alergiki se hrani, ki pri njih izzove reakcijo, izogibajo. Da je izogibanje uspešno in je hrana varna za vsakega posameznika, je pomembno dosledno in enotno označevanje alergenov. Označevati je treba predpakirana živila, nepredpakirana živila in jedi v gostinskih obratih. Med te spadajo tudi planinske koče, ki so po Zakonu o gostinstvu dolžne označevati alergene. Z ogledom jedilnikov na kočah in anketnim vprašalnikom za upravnike koč smo preverili pravilnost označevanja jedilnikov in poznavanje alergenov s strani upravnikov planinskih koč. Dobljene podatke smo statistično analizirali s Kruskal-Wallisovim testom. Oblikovali smo faktor poznavanja alergenov upravnikov glede na pravilne in nepravilne odgovore in jih analizirali glede na štiri neodvisne spremenljivke: starost, izobrazbo, izkušnje in kategorijo koče. Posebej smo analizirali tudi pravilnost vsakega odgovora glede na štiri neodvisne spremenljivke. Pokazali smo, da mlajše osebje bolje pozna problematiko alergenov. Raziskava je pokazala, da je treba osebje na planinskih kočah bolje usposobiti za poznavanje alergenov in s tem za zagotavljanje varne hrane vsem potrošnikom. Food contains specific substances, namely proteins, to which the body may react with sensitization. This reaction can be mild, such as stomach cramps or diarrhoea, or life-threatening in the form of an anaphylactic shock. People with allergies avoid the food which causes their allergies to flare up. In order to ensure they are able to do so, and that the food is safe for each individual, it is important that allergens are consistently and uniformly marked. Pre-packaged foodstuffs, non-pre-packaged foodstuffs and dishes in hospitality operations must be marked. Hospitality operations include mountain cottages which are required by the Hospitality Industry Act to mark allergens. By inspecting the menus at mountain cottages and distributing questionnaires among the keepers of mountain cottages, we checked whether the menus are adequately marked and whether the keepers of mountain cottages know and understand allergens. We statistically analysed the obtained data with the Kruskal-Wallis test. We formed a factor of understanding allergens based on correct and incorrect answers, which we analysed by considering four independent variables. We also specifically analysed the correctness of each answer by considering the four independent variables. We proved that younger people are more familiar with the allergen issue. Our research shows that the staff of mountain cottages should be better trained in understanding allergens in order to ensure the provision of safe food for all consumers.

Cilj istraživanja: Cilj ovog rada je upoznavanje sa alergijskim kontaktnim dermatitisom kao jednom od mogućih nuspojava kozmetičkih pripravaka te sastavnicama kozmetičkih pripravaka koje ga najčešće uzrokuju. Ljekarnik je često prvi zdravstveni radnik kojemu se pacijent obrati za savjet. Stoga su znanja o mogućim alergenima u kozmetičkim pripravcima nužna pri skrbi i savjetovanju bolesnika i korisnika, bilo da se radi o prepoznavanju alergijskog kontaktnog dermatitisa ili izboru prikladnih pripravaka za prevenciju ili pri liječenju. Materijali i metode: Literatura se pretraživala prema temi istraživanja, predmetu istraživanja, autorima i časopisu. Pretraživala se od općih prema specijaliziranim člancima pri čemu su se odabrali članci relevantni za problematiku ovoga specijalističkog rada. Relevantni članci proučavali su se na analitički i kritički način s obzirom na definiranje znanstvenog i/ili stručnog problema, istraživanje postojećih znanja o definiranom problemu (literaturni navodi), oblikovanje radne hipoteze, odabir metoda za ispitivanje hipoteze, prikaz i analizu rezultata te izvedene zaključke. Rezultati: Jedna od mogućih nuspojava kozmetičkih pripravaka je alergijski kontaktni dermatitis koji je tema ovog specijalističkog rada. Kontaktni alergijski dermatitis (dermatitis allergica e contactu) upalna je dermatoza koja nastaje kao posljedica kasne alergijske reakcije preosjetljivosti tipa IV po Coombsu i Gellu. Na pojavu kontaktnog alergijskog dermatitisa utječu različiti čimbenici kao što je senzibilizirajući potencijal kontaktnog alergena, oštećena zaštitna barijera kože, učestalost izloženosti alergenu, koncentracija alergena, stanje imunosnog sustava. Alergijski kontaktni dermatitis na kozmetičke pripravke čini 2-4% svih posjeta dermatologu. Najčešći alergeni u kozmetičkim pripravcima su mirisi i konzervansi, ali potencijalni alergen može biti bilo koja sastavnica kozmetičkog pripravka. U radu su obrađene sastavnice kozmetičkih pripravaka koji su prepoznati kao najčešći alergeni i koji su dio standardne europske serije kontaktnih alergena prilikom epikutanog testiranja: mirisi, konzervansi, peruvijanski balzam, lanolin, p-fenilendiamin. Zaključak: Unatoč kontroli proizvodnje kozmetičkih pripravaka i ograničenjima primjene određenih sastojaka, alergijski kontaktni dermatitis uzrokovan sastavnicama kozmetičkih pripravaka čest je i rastući problem u općoj populaciji koji može značajno narušiti kvalitetu života pacijenta. Najbolja prevencija je izbjegavanje ponovnog kontakta sa alergenom. Bolesnike alergične na određene sastavnice kozmetičkih pripravaka treba upozoriti na potrebu pažljivog čitanja deklaracije o sastavu pojedinoga pripravka, te informirati o dostupnosti baza podataka sastojaka i potencijalnih alergena u kozmetičkim pripravcima. S obzirom na postojanje križne reaktivnosti, osobi preosjetljivoj na određeni alergen savjetovati koje su i druge tvari na koje potencijalno može biti alergična, i koje bi bilo najbolje izbjegavati. Objectives: The objective of this paper is to introduce allergic contact dermatitis as one of the possible side effects of cosmetic products and ingredients of cosmetic products that commonly cause allergic contact dermatitis. A pharmacist is often the first health worker whom patients ask for advice. Therefore, knowledge of possible allergens in cosmetic products is necessary for the care and counseling of patients and users, whether it is recognition of allergic contact dermatitis or the choice of suitable preparations for prevention or treatment. Materials and methods: The literature search was performed by topic of research, case studies, the authors and the journal. The search was conducted from general to more specialized articles and articles relevant for this work were selected. Relevant articles were studied analytically and critically with regard to the definition of a scientific and /or technical problem, the research of existing knowledge on a defined problem (literary citations), the formulation of a work hypothesis, the selection of hypothesis testing methods, the presentation and analysis of the results and the derived conclusions. Results: One of the possible side effects of cosmetics is allergic contact dermatitis which is the subject of this work. Contact allergic dermatitis (dermatitis allergica e contactu) is an inflammatory dermatosis resulting from the late allergic type IV hypersensitivity classified by Coombs and Gell. The appearance of contact allergic dermatitis depends on various factors such as sensitizing potential of contact allergen, damaged skin barrier, allergen exposure frequency, allergen concentration, and immune system status. Allergic contact dermatitis caused by cosmetics accounts for 2% to 4% of all visits to the dermatologist. Fragrances and preservatives are the most common cause of allergy to cosmetics, but all ingredients should be considered as potential sensitizers. This paper deals with ingredients of cosmetic products that are recognized as the most common allergens and are a part of the European standard patch test series: fragrances, preservatives, balsamum peruvianum, lanolin, p-phenylenediamine. Conclusion: Despite strict and regulated production of cosmetics and restrictions on the use of certain ingredients, allergic contact dermatitis caused by components of cosmetics is a common and growing problem in the general population that can significantly impair the quality of life of the patient. The best prevention is avoidance of contact with an allergen. Patients who are allergic to certain components of cosmetics should be warned about the need for carefully reading the cosmetic label of the ingredients and be given information on databases of cosmetic ingredients and potential allergens. Considering the existence of cross reactivity, a person who is sensitive to a particular allergen should be advised about other substances they may be potentially allergic to and which they should avoid.

Astma je kronično vnetje dihalnih poti zaradi alergije, virusnih infekcij dihal in dražilnih snovi v zraku. Število obolelih z astmo in alergijskimi bolezni vedno bolj narašča, predvsem pri otrocih. Izpostavljenost ploda in zgodnja življenjska doba sta še posebej pomembna pri razvoju astme in alergijskih bolezni. Menijo, da prehranjevanje matere med nosečnostjo in potem prehranjevanje otroka v zgodnjem otroštvu, ne vplivajo kritično na razvoj in potek bolezni. Namen diplomskega dela je ugotoviti ali prehrana v zgodnjem življenju vpliva na razvoj astme in alergijskih bolezni.Iskanje literature je potekalo po vnaprej zastavljenih vključitvenih in izključitvenih kriterijih, glede na zastavljeno raziskovalno vprašanje: »Kako prehrana v zgodnjem življenju vpliva na tveganje za razvoj astme in alergijskih bolezni?«S pomočjo analize strokovne literature smo ugotovili, da obstajajo številne raziskave, ki menijo, da materinska prehrana igra pomembno vlogo na razvoj astme pri otrocih. Dokazi kažejo, da sta ravnovesje hranil v obdobju fetusa ter raznovrstna prehrana v zgodnjem življenju, še posebej pomembna za kratkoročno in dolgotrajno zdravje. Strokovnjaki v času nosečnosti ne priporočajo izogibanje določeni hrani, kot so arašidi, orehi in školjke, razen pri materah, ki so same astmatiki, ker omenjena hrana ne poveča možnosti nastanka alergij pri otrocih.Zaradi povečane incidence astme in alergijskih bolezni, predvsem pri otrocih, poudarjamo na pomen in vpliv raznovrstne prehrane, v nosečnosti pri materah ter polno dojenje otrokov prvih šest mesecev starosti. Dietne omejitve med nosečnostjo in kasneje med dojenjem se niso pokazale učinkovitim v preventivi razvoja alergij pri otroku. Asthma is a chronic inflammation of the respiratory tract because of allergies, virus infections of the respiratory tract and other stimulus particles in the air. The number of the patients with asthma and allergic diseases are increasing very rapidly, especially children. Exposition of fetus and early life period is especially important for the development of asthma and allergic diseases. Experts suppose, that the food that mothers eat during pregnancy and food their child eats later in the early childhood is not a determining influence on the development and course of the illness. The purpose of the final work is to determine whether the food in the first years of a child influences the development of asthma and allergic disease.In the final part there is a narrative review of the professional and scientific literature. The search for literature was based on predetermined inclusion and exclusion criteria, according to the research question: “How does early childhood With help of analysing professional literature we realised that mother’s food plays an important role on developing asthma on children. Evidence shows facts that balanced nutrients in the fetal period and food in the first years is very important for short-term and long-term health. Experts non recommend avoiding certain food during the pregnancy, like peanuts, nuts and shellfish because it not increases the chance for developing childhood allergiesnutrition affect the risk of developing asthma and allergic diseases?”Due to the increased incidence of asthma and allergic diseases, especially in children, we emphasize the importance and impact of diverse nutrition, pregnancy and full breastfeeding of children for the first six months of age. Diet restrictions during pregnancy and later during breastfeeding have not been shown to be effective in preventing the development of childhood allergies.

Cilj: Objasniti zašto postoje serije alergena te pregledno prikazati one serije koje se koriste u Dječjoj bolnici Srebrnjak u Zagrebu. Opisati izradu epikutanih testova u bolničkoj ljekarni i raspraviti probleme vezane uz taj postupak. Prikazati i povezati različite nomenklature alergena Europske bazične serije i drugih serija koje se primjenjuju u Dječjoj bolnici Srebrnjak. Nakon završenog testiranja i očitanja rezultata pomoći pacijentima da nauče kako izbjegavati alergene na koje imaju dokazanu preosjetljivost. Materijal i metode: Pretraživala se literatura u bibliografskim bazama podataka (PubMed, Ovid) prema osnovnim ključnim riječima. Članci prikupljeni pretraživanjem razvrstani su od općih prema specijaliziranim pri čemu su odabrani članci relevantni za ovo istraživanje te su proučeni na analitički i kritički način. U radu su navedeni najvažniji rezultati i saznanja, kao i vlastita razmatranja. Rezultati: U Republici Hrvatskoj je najvažnija europska bazična serija (EBS) koja se i najčešće koristi za izvođenje bolničkih alergijskih testova. Bazične serije se stalno ažuriraju i objavljuju se kriteriji za uključivanje novih alergena koji postaju rizični za određenu populaciju. Probirne serije haptena se koriste u kombinaciji sa bazičnim serijama da bi se olakšalo testiranje u pacijenata kod kojih postoji sumnja na izloženost alergenima uslijed njihove profesije ili iz neke druge specifične okoline. Razmotrena je uporaba različitih koncentracija pri izradi epikutanih testova na lijekove. Izrađena je lista svih alergena koji se koriste za epikutano testiranje u Dječjoj bolnici Srebrnjak na kojoj se nalaze njihovi brojni sinonimi i pregled mogućih izvora tih alergena koje pacijenti s dokazanom preosjetljivošću moraju izbjegavati. Zaključak: Postojanje nacionalnih i internacionalnih bazičnih serija haptena i, uz njih, probirnih serija razrađenih po profesijama i drugim kriterijima vrlo je važno jer su one stvorene prema posebnim rizicima karakterističnim za svako geografsko područje i druga područja izloženosti, naročito profesionalna. Izrada bolničkih epikutanih testova može biti od velike pomoći u dijagnostici specifične populacije, a pri tom procesu važno je koristiti smjernice Europskog društva za kontaktni dermatitis te razviti metode izrade koje će imati što manje fluktuacija u koncentracijama djelatnih tvari. Na kraju cijelog procesa dijagnostike vrlo je važno pacijentima pružiti pisane podatke i upute o izbjegavanju „njihovih“ alergena. Objectives: Explain why there are a series of allergens and clearly show a series that is being used at the Children's Hospital Srebrnjak in Zagreb. Describe the production of patch tests at a hospital pharmacy and discuss the problems associated with this procedure. Display and link different nomenclatures of the European baseline series allergen and other series applied at the Children's Hospital Srebrnjak. After completing the test and reading the results help patients learn how to avoid allergens with proven hypersensitivity. Material and Methods: Literature was searched in bibliographic databases (PubMed, Ovid) according to basic keywords. The articles collected by searching are classified from general to specialized, with selected articles relevant to this research and analyzed in an analytical and critical manner. The paper presents the most important results and findings as well as my own deliberations. Results: For us, the most important is European baseline series (EBS) which is used in the Republic of Croatia. Baseline series is constantly updated and criteria are published for the inclusion of new allergens that become risky for a particular population. Hapten screening series are used in combination with baseline series to facilitate testing in patients suspected of allergens from their profession or other specific environment. Different concentrations of patch drug tests were considered. A list of allergens used for patch testing in the Children's Hospital Srebrnjak is made, which contains their numerous synonyms and an overview of possible sources of these allergens that patients with proven hipersensitivity have to avoid. Conclusion: The existence of national and international baseline hapten series and, beside them, screening series composed according to occupations and other criteria is very important because they are created according to the special risks characteristic of each geographic area and other areas of exposure, particularly professional. The production of hospital patch tests can be of great help in the diagnosis of a specific population and during this process it is important to use the European Society for Contact Dermatitis guidelines and develop methods of production that will have as little fluctuation as possible in concentrations of active substances. At the end of the entire diagnostic process, it is very important for patients to provide written information and instructions on avoiding "their" allergens.

Tijekom ovog istraživanja određeni su potencijalni alergeni kikirikija u trideset i devet uzoraka sladoleda prikupljenih u slastičarnicama i trgovačkim lancima pomoću ELISA (engl. Enzyme-Linked Immunosorbent Assay) metode te su prikupljeni stavovi i mišljenja potrošača o kikirikiju i proizvodima od kikirikija kao tvarima ili proizvodima koji uzrokuju alergije. Alergeni kikirikija detektirani su u petnaest od ukupno devetnaest uzoraka sladoleda iz slastičarnica, a udio se kretao u rasponu od 0,15 mg kg-1 do 12,22 mg kg-1. Između dvadeset analiziranih uzoraka sladoleda iz trgovačkih lanaca, alergeni kikirikija detektirani su kod njih dvanaest, a udio je iznosio od 0,20 mg kg-1 do 5,44 mg kg-1. Rezultati određivanja alergena kikirikija imunoenzimskom ELISA metodom uspoređeni su također sa informacijama o hrani analiziranih uzoraka, a vezanim uz prisutnost alergena kikirikija, te se podudaraju u četiri slučaja od devetnaest nepretpakiranih uzoraka sladoleda iz slastičarnica i dvanaest od dvadeset zapakiranih uzoraka sladoleda iz trgovačkih lanaca. Većina ispitanika (84,4 %) smatra da sladoled može sadržavati tragove kikirikija ukoliko kikiriki nije jedan od sastojaka sladoleda, a obzirom na dodatne navode o mogućoj prisutnosti alergena kikirikija, 40,1 % ispitanika smatra da proizvod kao što je sladoled ne sadrži tragove kikirikija ukoliko na deklaraciji nema dodatnog navoda o njegovoj mogućoj prisutnosti. During this research, potential peanut allergens were detected in thirty-nine ice cream samples collected from pastry shops and supermarkets using the ELISA (Enzyme-Linked Immunosorbent Assay) method, and the views and opinions of consumers about peanuts and peanut products as substances or products that cause allergies was examined. Peanut allergens were detected in fifteen of nineteen ice cream samples, and the content ranged from 0.15 mg kg-1 to 12.22 mg kg-1. Between twenty analysed ice cream samples, in twelve peanut allergens were detected, and the content was between 0.20 mg kg-1 to 5.44 mg kg-1. The results of peanut allergen determination by the immunoenzymatic ELISA method were also compared with the food information of the analysed samples, related to the presence of peanut allergens, and matched in four cases of nineteen non-prepacked ice cream samples from pastry shops and twelve out of twenty packed ice cream samples from supermarkets. Most respondents (84.4%) thought that ice cream could contain traces of peanuts if peanuts were not one of the ingredients of ice cream, and given the additional allegations of possible presence of peanut allergens, 40.1% of respondents thought that a product such as ice cream did not contain traces of peanuts if there is no additional indication of its possible presence on the declaration.

Alergija na hrano je potencialno smrtno nevarna imunska reakcija, ki jo običajno sprožijo neškodljivi antigeni proteinov hrane. Razširjenost alergij na hrano raste, do danes pa ne obstaja nobena odobrena terapija zdravljenja razen izogibanja alergena hrane ter zdravljenja akutnih alergijskih reakcij z vbrizganjem epinefrina. Oralna imunoterapija predstavlja obetajočo terapevtsko alternativo, kjer pacienti uživajo majhne, a postopoma večje količine alergena tekom več mesecev, vendar je povezana s pogostimi alergijskimi reakcijami. Medtem ko je desenzibilizacija ali zmanjšana reaktivnost na alergen hrane, dosežena pogosto, indukcija dolgotrajne tolerance še ni bila vzpostavljena. V delu se bom osredotočila na glavne dosežke in nedavne napredke v oralni imunoterapiji za zdravljenje pogostih alergij na hrano, kot so arašidi, kravje mleko, jajca in pšenica. Food allergy is potentially life-threatning immune reaction, triggered by normally innocous food protein antigens. The prevalence of food allergy is rising, but there is no approved therapy apart from avoidance of allergenic foods and treatment of acute allergic reactions with injectable epinephrine. Oral immunotherapy is a promising therapeutic alternative, in which patients consume small, but gradually increasing alergen doses over many months, but it is associated with frequent allergic reactions. While desensitization, or reduced reactivity to food alergens, is often achieved, the induction of long term tolerance has not yet been established. In this work I will focus on main acchivements and recent progres in oral immunotherapy for treatment of common food alergies, such as peanut, cow's milk, hen's egg and wheat.

Tijekom ovog istraživanja određeni su potencijalni alergeni lješnjaka u sedam istovrsnih uzoraka čokolada s različitim datumom proizvodnje i sedam različitih uzoraka čokoladnih proizvoda pomoću ELISA (engl. Enzyme-Linked ImmunoSorbent Assay) metode te je ispitan stav potrošača o navodima potencijalne prisutnosti alergena lješnjaka na deklaracijama ispitivanih proizvoda. U svim analiziranim uzorcima detektirana je prisutnost alergena lješnjaka, a određeni udjel nije prelazio >20 mg kg-1. Između analiziranih uzoraka u kojima su određeni alergeni lješnjaka, svi proizvodi su sadržavali navod o potencijalnoj prisutnosti alergena lješnjaka na deklaraciji. Iako samo mali broj ispitanika čita deklaracije na proizvodima, utvrđeno je da navod „Može sadržavati tragove lješnjaka“ 63,5 % ispitanika izdvaja kao najpoznatiji. U ovom istraživanju, u šest analiziranih istovrsnih čokoladnih uzoraka i tri uzorka različitih čokoladnih proizvoda, ELISA metodom analizirani su alergeni lješnjaka. During this research potential hazelnut alergens are defined in seven of the same chocolate samples with different date of production and seven different chocolate samples using ELISA (Enzyme-Linked ImmunoSorbent Assay) method and consumer attitude on allegations of potential presence of hazelnut allergens on declarations of the tested products was examined.. In all analyzed samples hazelnut allergens were detected, but determined content did not exceed amount of >20 mg kg-1. Between analyzed samples in which hazelnut allergens are determinated, all products contained a claim on potential presence of hazelnut allergens on declaration. Although only a small number of respondents read product declarations, it was found that the quote „May contain traces of hazelnuts“ 63,5% of respondents claim as best known. In this research, in six of the same analyzed chocolate samples and three samples of different chocolate products, hazelnut allergnes are analyzed by ELISA method.

Cilj ovog istraživanja bio je odrediti udio potencijalnih alergena kikirikija imunoenzimskom ELISA metodom (engl. Enzyme-Linked ImmunoSorbent Assay) u 8 uzoraka sladoleda od čokolade, odnosno četiri uzorka sladoleda iz različitih slastičarnica te četiri uzorka sladoleda različitih proizvođača iz trgovačkih lanaca. Analitički rezultati uspoređeni su s navodima u okviru informacija o hrani vezanim uz prisustvo alergena. U sedam od ukupno osam analiziranih uzoraka sladoleda, detektirani su alergeni kikirikija. Raspon unutar kojeg se kretao udio alergena kikirikija u analiziranim uzorcima sladoleda od čokolade iz slastičarnica iznosi od 0,27 do 0,62 mgkg-1, dok se u uzorcima sladoleda iz trgovačkih lanaca udio kretao u rasponu od 0,12 do 0,35 mgkg-1. Najviši udio alergena kikirikija određen je u uzorku ''SS4'' iz slastičarnice te iznosi 0,62 mgkg- 1, pri čemu je u okviru informacija o hrani naveden sadržaj alergena među kojima nije istaknut kikiriki. The purpose of this study was to determine the content of potential peanut allergens by using the immunoenzyme ELISA method (Enzyme-Linked ImmunoSorbent Assay) in eight samples of chocolate ice cream, from which four of samples were from different pastry shops, and the other four were ice creams of different manufacturers from a retail chain. The analytical results were compared with the food information claims related to the presence of allergens. Peanut allergens were detected in seven out of eight analysed samples. In samples of chocolate ice cream from pastry shops, the range of determined peanut allergens was from 0,27 to 0,62 mgkg-1, and in samples from a retail chains from 0,12 to 0,35 mgkg-1. The highest content of peanut allergens was 0,62 mgkg- 1 and it was determined in a sample from a pastry shop ''SS4'' which contained an advisory label related to the presence of allergens, but didn't contain a claim related to the presence of peanut.

Tijekom ovog istraživanja, imunoenzimskom ELISA (engl. Enzyme-Linked Immunosorbent Assay) metodom analizirani su udjeli potencijalnih alergena kikirikija u 28 uzoraka keksa i čajnih peciva iz trgovačkih lanaca, pekarnica i slastičarnica te su prikupljeni stavovi i mišljenja potrošača o kikirikiju i proizvodima od kikirikija kao tvarima ili proizvodima koji uzrokuju alergije. U svih deset analiziranih uzoraka keksa i čajnih peciva iz trgovačkih lanaca detektirani su alergeni kikirikija, a prosječni udio iznosio je 0,37 mgkg-1. Alergeni kikirikija detektirani su u devet od deset uzoraka keksa i čajnih peciva iz pekarnica, a njihov udio iznosio je prosječno 0,38 mgkg-1. Prosječni udio alergena kikirikija, detektiranih u osam analiziranih uzoraka keksa i čajnih peciva iz slastičarnica, iznosio je 0,28 mgkg-1. Rezultati ispitivanja stavova i mišljenja potrošača pokazali su da je 84,8 % ispitanika upoznato sa pojmom alergije te također 39,2 % ispitanika smatra kako se kikiriki ubraja u orašaste plodove. Od dvadeset i osam analiziranih uzoraka keksa i čajnih peciva, ELISA metodom detektirani su alergeni kikirikija u dvadeset sedam uzoraka pri čemu su samo dva uzorka iz trgovačkog lanca sadržavala navod o mogućoj prisutnosti alergena kikirikija. During this study, the content of potential peanut allergens in 28 samples of cookies and tea biscuits from supermarkets, bakeries and pastry shops was analysed by ELISA (Enzyme-Linked Immunosorbent Assay) method, and also the consumers opinion on peanut and peanut products as substances or products that cause allergies was examined. The presence of peanut allergens was detected in ten analysed samples of cookies and tea biscuits from supermarkets, with an average value of 0,37 mgkg-1. Peanut allergens were detected in nine of ten samples of cookies and tea biscuits from bakeries, with an average value of 0,38 mgkg-1. The average content of peanut allergens, detected in eight analysed samples of cookies and tea biscuits from pastry shops, was 0,28 mgkg-1. The results of consumers attitudes and opinions showed that 84,8 % of respondents are familiar with the term of allergy and also 39,2 % of respondents consider peanuts as tree nuts. Among twenty eight analysed samples of cookies and tea biscuits by ELISA method, peanut allergens were detected in twenty seven samples of which only two from supermarkets had advisory label related to the presence of peanut allergens.

Nezaželene snovi v hrani, denimo alergeni in toksini, so nam poznani že dlje časa. Število ljudi, ki so nagnjeni k alergijam na hrano, iz leta v leto narašča. Zato smo se s časom določene hrane naučili izogibati ali jo procesirati na določene načine, da nam je postala užitna in smo se tako izognili alergijskim reakcijam ter drugim nevšečnostim. Da se izognemo tveganju, smo do nedavnega izvajali tudi izboljšave rastlin s selektivnim križanjem in mutacijami ter na ta način iskali sorto, ki bi morda ustrezala zastavljenim ciljem. V sodobnem času to rešujemo drugače, denimo z novimi biotehnološkimi metodami, ki nam omogočajo modifikacijo specifične regije v genomu. Z njimi lahko nezaželene snovi izločimo, tako da izločimo njihov gen ali pa preprečimo njegovo transkripcijo. Z RNAi metodo je znanstvenikom uspelo doseči kar nekaj prepričljivih rezultatov, vendar so zaradi boljše učinkovitosti, nižje cene in predvidene lažje sprejemljivosti v javnosti, danes v ospredju metode ZFN, TALEN in CRISPR/Cas9. Primerov izločitev nezaželenih snovi s temi metodami je manj, vseeno pa obstaja kar nekaj prepričljivih dosežkov izzvane tarčne modifikacije. Omenjena je tudi problematika GSO po svetu. Predvidevam, da se bo v prihodnosti uporaba biotehnoloških metod zaradi efektivnosti še povečala, genski inženiring pa bo kmalu bolje sprejet tudi s strani javnosti. Nondesired substances in food such as allergens and toxins have been known to us for many years. The number of people that are prone to allergies of certain foods is increasing from year to year. Therefore, we have learnt to avoid or to process them in certain ways in order to prevent allergic reactions and other inconveniences. Until recently the improvements of plants with selective breeding and mutations and searching for an appropriate variety was also in progress. Nowadays this is solved by new biotechnological methods which allow us to mutate specific region in the genome. These methods eliminate unwanted substances with elimination of their gene or prevention of transcription. With RNAi method scientists in this field managed to reach quite a few impressive accomplishments, but nowadays ZFN, TALEN and CRISPR/Cas9 methods are becoming more popular because of better effectiveness, lower prices and desired public acceptance. Scientists managed to make some impressive achievements of induced target mutagenesis. We assume that in the future the use of genetic engineering will increase because of its effectiveness and genetic engineering will soon become better accepted by general public.

Kontaktni dermatitis je upala kože koja nastaje nakon kontakta s tvarima koje mogu nadražiti kožu ili izazvati alergijsku reakciju. Praćen je svrbežom, osipom, crvenilom, otokom, a mogu se javiti i bule koje pucaju te kruste koje predstavljaju sasušeni sadržaj puknutog vezikula. Razlikujemo sljedeće tipove kontaktnog dermatitisa: kontaktni nealergijski dermatitis (akutni i kronični), kontaktni alergijski dermatitis (akutni i kronični), fotoalergijski kontaktni dermatitis, fototoksični kontaktni dermatitis te sindrom kontaktne urtikarije. Kliničke su slike različite. Broj oboljelih u stalnom je porastu. Kontaktni nealergijski odnosno iritativni dermatitis kožna je upala koja nastaje nakon izlaganja koncentriranim lužinama i kiselinama prilikom čega se oštećuje gornji sloj epidermisa bez sudjelovanja imunosnih mehanizama. Kontaktni alergijski dermatitis upalna je dermatoza koja nastaje kao posljedica alergijske reakcije tipa IV, posredovane limfocitima. Podjednako se pojavljuje kod muškaraca i žena. Temeljni test za dokaz alergijske preosjetljivosti je epikutani ili patch test. U dijagnostici je također bitna klinička slika te anamneza. Jačina upale strogo je promjenjiva te ovisi o nizu različitih čimbenika kao što su količina i snaga određenog iritansa ili alergena, duljina i frekvencija izloženosti, okolišni čimbenici itd. Izuzetno je važno izbjegavati kontakt s uzročnim alergenom i upotrebljavati zaštitna sredstva. Profesionalni kontaktni dermatitis može biti alergijske i nealergijske geneze, a nastaje kao posljedica kontakta kože s različitim tvarima u radnoj sredini. U nastanku profesionalnog dermatitisa bitni su podatci o vrsti posla i tvarima u radnome prostoru, o uvjetima rada, stanju kože prije zaposlenja, o primjeni zaštitne odjeće i drugih zaštitnih sredstava. Ukoliko se kontakt s inkrimiranom tvari na radnom mjestu ne može izbjeći, poželjna je promjena radnog mjesta. Contact dermatitis (CD) is skin inflammation that occurs after contact with substances that can irritate skin or cause allergic reaction. It is characterised by itch, rash, redness, swelling and sometimes there can also be vesicles that burst and crusts with content of broken blister. According to etiological division there are different types of contact dermatitis: contact non-allergic dermatitis (acute and chronic), contact allergic dermatitis (acute and chronic), photoallergic contact dermatitis, phototoxic contact dermatitis and contact urticaria syndrome. Clinical features are different. Number of people suffering from CD is increasing. Contact non-allergic or irritative dermatitis is a skin inflammation that occurs after exposure to concentrated alkalis and acids which cause damage to the upper layer of epidermis, without immune mechanisms. Contact allergic dermatitis is an inflammatory skin disease produced by IV type allergic reaction, mediated by lymphocytes. It is equally distributed among males and females. The most important diagnostic methods for allergic hypersensitivity are: anamnesis, clinical picture and skin patch test. The intensity of inflammation depends on many different factors: amount and strenght of the irritant, length and frequency of the exposure, environmental factors etc. It is extremely important to avoid any contact with the allergen and use protective gloves, masks and clothings. Professional contact dermatitis can be of allergic and non-allergic origin, and it is caused by skin contact with many different substances in the working environment. In the development of professional dermatitis informations about profession, potential alergens or irritant, substances in the working place, working conditions, skin condition before starting specific job, protective clothing and other protective things are very important. If allergens cannot be removed or avoided the change of job position or professional retraing might be needed.

Kikiriki se ubraja u najčešće nutritivne alergene koji kod alergičnih osoba može izazvati teške reakcije. Cilj ovog istraživanja bio je odrediti udio potencijalnih alergena kikirikija u šesnaest uzoraka kolača koji sadrže kakaove dijelove pomoću ELISA testa te prikupiti stavove i mišljenja potrošača o kikirikiju i proizvodima od kikirikija kao tvarima ili proizvodima koji uzrokuju alergije. Alergeni kikirikija detektirani su u analiziranim uzorcima nepretpakiranih kolača iz slastičarnice, a njihov udio iznosio je prosječno 0,64 mgkg-1. Prosječni udio alergena kikirikija u analiziranim uzorcima kolača iz objekta javne prehrane iznosio je 0,13 mgkg-1, a u uzorcima iz trgovačkog lanca 0,62 mgkg-1. Većina ispitanika (82,9 %) upoznata je s pojmom alergije, no njihova percepcija o tome u koju skupinu alergena se ubraja kikiriki često je pogrešna pa mnogi (57,9 %) smatraju kako se kikiriki ubraja u orašaste polodove. Od šesnaest ukupno analiziranih uzoraka nepretpakirane hrane, u jedanaest uzoraka bez navoda o prisutnosti kikirikija, ELISA metodom detektirani su alergeni kikirikija. Peanut is one of the most common food allergens which in allergic people can cause life- threatening hypersensitivity reactions. In this study the commercial ELISA (Enzyme-Linked ImmunoSorbent Assay) test was used for determination of potential peanut allergens in sixteen samples of nonpacked cakes with cocoa parts and also the consumers opinion on peanut and peanut products as substances or products that cause allergies was examined. Peanut allergens were detected in analyzed samples of nonpacked cakes from pastry shop with average value of 0,64 mgkg-1. The average value of peanut allergens in the analyzed cake samples from the public eating facility was 0,13 mgkg-1 and from supermarket 0,62 mgkg-1. Most of the respondents (82,9 %) are familiar with term 'allergy' but their perception about peanut is often wrong and many (57,9 %) consider that it belongs to nuts. In this study, among eleven samples of nonpacked cocoa cakes, which had no advisory label, peanut allergens were detected by ELISA method.

Tijekom ovog istraživanja prisutnost alergena kikirikija je analizirana ELISA (Enzyme-linked immunosorbent assay) metodom u uzorcima čokolade s lješnjakom i krem-proizvoda iz trgovačkih lanaca te uzorcima kolača koji sadrže kakaove dijelove iz trgovačkog lanca, slastičarnice i objekta javne prehrane, a dobiveni rezultati uspoređeni su sa informacijama o hrani vezanim uz prisutnost alergena kikirikija. U petnaest od šesnaest analiziranih uzoraka zapakirane hrane (čokolada s lješnjakom i krem-proizvodi) detektirani su alergeni kikirikija pri čemu je navod o sadržaju alergena kikirikija, u okviru informacija o hrani, sadržavalo šest takovih proizvoda. Alergeni kikirikija detektirani su i u četrnaest od šesnaest analiziranih uzoraka nepretpakirane hrane, odnosno kolača koji sadrže kakaove dijelove iz trgovačkog lanca, slastičarnice i objekta javne prehrane pri čemu su informacije o hrani ukazivale na sadržaj kikirikija u tri takova proizvoda. Rezultati istraživanja ukazuju na značaj detekcije alergena kikirikija kao skrivenog sastojka u hrani, obzirom da je podatke o tvarima ili proizvodima koji uzrokuju alergije ili netolerancije koji između ostalog uključuju kikiriki i proizvode od kikirikija, potrebno navoditi u okviru obveznih informacija o hrani.

Cilj ovog rada je utvrditi prisutnost sljedećih alergena: glutena, soje, mlijeka i gorušice u različitim vrstama mesnih proizvoda te istražiti sukladnost sa oznakama na proizvodu odnosno udovoljavanje zahtjevima zakonodavstva s obzirom na utvrđene količine. Ispitivanje je provedeno na 51 uzorku mesnih proizvoda proizvedenih od strane različitih mesnih industrija i zastupljenih na hrvatskom tržištu. Količina alergena (mg/kg) određivana je primjenom validiranih ELISA (enzyme-linked immunosorbent assay) metoda. Rezultati istraživanja pokazuju da je najveća količina glutena prisutna u hrenovkama (63,7 mg/kg), proteina mlijeka (380,4 mg/kg) i soje (25,7 mg/kg) u paštetama te gorušice (14,0 mg/kg) u kobasicama za pečenje. U šest mesnih proizvoda (11,8%) određeni su alergeni veći od limita kvantifikacije (LOQ) primjenjenih analitičkih metoda, a da njihova prisutnost nije bila označena. Rezultati istraživanja upućuju na važnost sustavne kontrole alergena u mesnim proizvodima u svrhu zaštite zdravlja i interesa potrošača., The goal of this paper is to establish the presence of the following allergens: gluten, soy, milk and mustard in various types of meat products and research their compliance with the product labels, i.e. meeting the legal requirements with regard to the established quantities. The test was conducted on 51 meat product samples produced by different meat industries represented on the Croatian market. The quantity of the allergens (mg/kg) was defined by applying validated ELISA (enzyme-linked immunosorbent assay) methods. The research results show that the largest amount of gluten is present in frankfurters (63.7 mg/kg), milk protein (380.4 mg/kg) and soy (25.7 mg/kg) in pates and mustard (14.0 mg/kg) in grill sausages. In six meat products (11.8%), allergens have been established above the quantification limit (LOQ) of the applied analytical methods, while their presence was not indicated on the label. The research results point to the importance of systemic control of allergens in meat products in order to protect consumer interests and health., Ziel dieser Arbeit ist es, die Anwesenheit der Allergene Gluten, Soja, Milch und Senf in diversen Fleischprodukten festzustellen und die Übereinstimmung mit der Produktkennzeichnung bzw. mit den gesetzlichen Anforderungen in Abhängigkeit von den festgelegten Mengen zu überprüfen. Die Untersuchung wurde an 51 Fleischproben durchgeführt, die aus diversen Fleischbetrieben stammen und auf dem kroatischen Markt vertreten sind. Die Allergenen-Menge (mg/kg) wurde mittels validierter ELISA (enzyme-linked immunosorbent assay) Methoden festgelegt. Die Ergebnisse der Untersuchung zeigen, dass der höchste Gehalt von Gluten in Würstchen (63,7 mg/kg), von Milchproteinen (380,4 mg/kg) und Soja (25,7 mg/kg) in Leberpasteten und von Senf (14,0 mg/kg) in Bratwürsten festgestellt wurde. Bei sechs Fleischprodukten (11,8%) wurde ein Anteil von Allergenen ermittelt, der das Quantifikationslimit (LOQ) der angewandten analytischen Methoden übersteigt, ohne dass deren Gehalt ausgewiesen wurde. Die Untersuchungsergebnisse weisen auf die Bedeutung einer systematischen Kontrolle der Allergene in Fleischprodukten zum Schutz der Gesundheit und der Interessen der Verbraucher hin., Obiettivo di questo studio è accertare la presenza dei seguenti allergeni: glutine, soia, latte e senape in vari tipi di prodotti di carne e fare luce sulla conformità delle etichette sul prodotto, ossia la soddisfazione dei requisiti di legge rispetto alle quantità accertate. Il test ha interessato 51 campioni di prodotti di carne provenienti da diverse industrie alimentari e presenti sul mercato croato. La quantità di allergeni (mg/kg) è stata determinata applicando i metodi verificati ELISA (enzyme-linked immunosorbent assay). I risultati del test hanno dimostrato che il glutine è presente in maggior quantità nei wurstel (63,7 mg/kg), le proteine del latte (380,4 mg/kg) e della soia (25,7 mg/kg) nei pâté di carne e la senape (14,0 mg/kg) nelle salsicce da cuocere alla piastra. In sei prodotti di carne (11,8%) è stata accertata una presenza di allergeni superiore al limite di quantificazione (LOQ) dei metodi analitici applicati e che la loro presenza non era stata indicata nell’etichetta. I risultati della ricerca rimandano all’importanza dei controlli sistematici per individuare la presenza di allergeni nei prodotti di carne al fine di tutelare la salute e gli interessi dei consumatori., El objetivo principal de este trabajo es determinar la presencia de los siguientes alérgenos: gluten, soya y sinapis en diferentes tipos de productos cárnicos e investigar su correspondencia con los datos en las etiquetas de los productos, en otras palabras, el cumplimiento de los requisitos de la legislación con respecto a las cantidades determinadas. La investigación fue realizada en 51 muestras de productos cárnicos de varios productores de la industria cárnica presentes en el mercado croata. La cantidad de alérgenos (mg/kg) se determinó usando la validación de método ELISA (enzyme-linked immunosorbent assay). Los resultados demuestran que la mayor cantidad de gluten está presente en salchichas de tipo Frankfurt (63,7 mg/kg); la de proteínas lácteas (380,4 mg/kg) y soya (25,7 mg/kg) en patés; y la de sinapis, (14,0 mg/kg) en salchichas para freír. En seis productos cárnicos (11,8) ciertos alérgenos sobrepasan el límite de cuantificación (LOQ) de los métodos analíticos usados, mientras que su presencia no se especificaba en la etiqueta. Los resultados de investigación destacan la importancia del control sistemático de alérgenos en los productos cárnicos con el fin de proteger la salud y los intereses de los consumidores.

Cilj ovog rada bio je analizirati prisutnost alergena glutena, soje, proteina mlijeka i gorušice u različitim vrstama mesnih proizvoda te istražiti označavaju li proizvođači eventualno prisutne alergene u skladu sa važećom zakonskom regulativom. Određivanje alergena provedeno je primjenom ELISA metode na pedeset i jednom uzorku različitih mesnih proizvoda različitih proizvođača zastupljenih na tržištu Republike Hrvatske. Rezultati istraživanja pokazali su kako se najviša razina glutena nalazi u hrenovkama (63,73 mgkg-1), proteina mlijeka u paštetama (380,42 mgkg-1), soje također u paštetama (25,7 mgkg-1) i gorušice u kobasicama za pečenje (14 mgkg-1). U šest proizvoda određeni su alergeni, a da njihova prisutnost nije bila navedena na deklaraciji. Rezultati istraživanja upućuju na važnost sustavne kontrole alergena u mesnim proizvodima u svrhu zaštite zdravlja i interesa potrošača. The aim of this study was to determine the presence of allergens gluten, soy, milk and mustard in different samples of meat products and compare compliance beetween the informations on declarations and the current legislative. Determination of allergens was carried out with ELISA method on fifthy-one sample of different meat products from Croatian market. The results of study showed that the highest concentration of gluten were found in hot dog sausages (63,73 mgkg-1), milk in pâtés (380,42 mgkg-1), soy also in pâtés (25,7 mgkg-1) and mustard in grill sausages (14 mgkg-1). In six products, allergens were found but not declared on package. The results point out the importance of systematic analysis of meat products on market in order to improve protection of consumers health and interest.

Cilj ovog rada je dati pregled eksperimentalnih i kliničkih podataka o poznatim i potencijalno alergenim tvarima u vinarstvu, kao i trenutne zakonske regulative za takve tvari. Tijekom vinifikacije, grožđe, mošt i vino su osjetljivi na promjene i defekte koji bitno utječu na prihvatljivost kod potrošača s alergijama. Alergeni aditivi, kao sulfiti i lizozim iz jaja se primjenjuju radi kontrole procesa fermentacije i kvarenja vina. Biogeni amini nastali u visokim koncentracijama mogu direktno ili indirektno utjecati na toksičnost i alergijski potencijal vina. Proteinski proizvodi se koriste kao sredstva za bistrenje, za uklanjanje neželjenih netopljivih čestica i koloidnnih tvari mošta ili vina, radi poboljšanjea njihove stabilnosti i senzorskih svojstava. Neki od njih (bjelanjak, kazein, riblja želatina) imaju alergijski potencijal i prisutnost njihovih ostataka u konačnom proizvodu može predstavljati rizik za alergične osobe. The aim of this work was to review the experimental and clinical data about known and potentially allergenic substances in wine, as well as the current legislation for such substances. During vinification, grapes, grape must and wine are sensitive to changes and defects that materially affect the acceptability among consumers with allergies. Allergens additives, like sulfites and lysozyme, are included among the wine supplements in terms of control of the fermentation process and the perishability of wine. Biogenic amines in high concentrations can directly or indirectly influence the toxicity and allergic potential of wine. Proteinous products are widely used as fining agents in wine to remove unwanted insoluble particles and colloidal materials from the must or wine, and to improve stability organoleptic properties. Some of them (egg, casein, fish gelatin) have allergic potential and the presence of their residues in the final product may pose a risk for allergic people.