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The aim of the present study was to determine the frequency of different allergies in horses from central Poland. Allergic diseases are common in horses all over the world. Common conditions that have been associated with immunoglobulin E (IgE)-mediated allergy in horses include insect bite hypersensitivity (IBH), urticaria and recurrent airway obstruction (RAO). A variety of serological assays are available to measure serum levels of allergen-specific IgE antibodies. They can provide supportive evidence for a diagnosis of equine allergic dermatitis associated with environmental allergens. This study was performed on clinically healthy, Polish noble half-breed horses. In this study, four groups of allergens were identified as commonly inducing allergic reactions: American house dust mites (D. farinae), cultivated plants (rape), English plantain (Plantago lanceolata) and biting midges (Culicoides nubeculosus). More than 50% of horses showed positive reactions to these groups of allergens. The factor that contributed most to allergy occurrence among horses was American house dust mites (D. farinae). In recent years, efforts have been made to identify new allergens and to better understand the specific pathogenesis of allergic diseases in animals. The development of allergic disease diagnostics has been facilitated by the availability of diagnostic methods along with enhanced awareness of allergic diseases in veterinary practice. Allergic disorders in horses are ultimately gaining the attention they deserve thanks to the fact that veterinary allergology has become a dedicated veterinary discipline., Cilj ove studije bio je ustvrditi učestalost različitih vrsta alergija u konja iz središnje Poljske. Alergijske bolesti su učestale u konja diljem svijeta. Uobičajena stanja koja su povezana s alergijskom reakcijom posredovanom imunoglobulinom E (IgE) u konja uključuju preosjetljivost na ubode insekata (IBH), urtikariju i povratnu opstrukciju dišnih putova (RAO). Dostupni su različiti serološki testovi za mjerenje razina specifičnog IgE antitijela na alergene u krvi. Oni mogu osigurati potkrepljujuće dokaze za dijagnozu alergijskog dermatitisa konja s alergenima iz okoliša. Ova studija provedena je na poljskim plemenitim, klinički zdravim, križanim pasmina konja. U ovoj studiji identificirane su četiri skupine alergena koje obično prouzroče alergijske reakcije: američke grinje iz kućne prašine (D. farinae), kultivirane biljke (repica), uskolisni trputac (Plantago lanceolata) i mušice (Culicoides nubeculosus). Više od 50 % konja pokazalo je pozitivne reakcije na ove skupine alergena. Čimbenik koji je najviše pridonio pojavnosti alergijskih reakcija među konjima bile su američke grinje iz kućne prašine (D. farinae). U posljednjih nekoliko godina, pojačanje napora u suzbijanju alergijskih bolesti dovelo je do identifikacije nekih novih alergena, kao i dubljeg razumijevanja patogeneze životinja specifične za alergijske bolesti. Razvoj u dijagnozi alergijskih bolesti olakšan je dostupnošću dijagnostičkih metoda uz povećanu svijest o alergijskim bolestima u veterinarskoj praksi. Alergijski poremećaji u konja konačno dobivaju pozornost koju zaslužuju zahvaljujući činjenici da je alergologija postala značajna specijalizirana veterinarska disciplina.

Alergije na hranu su imunološki posredovane reakcije organizma i bolesti koje se ponavljano javljaju nakon ingestije određene hrane. Predstavljaju značajan javnozdravstveni problem, naročito u ekonomski razvijenim zemljama. Češće su u dječjoj nego u odrasloj dobi, a mogu zahvatiti respiratorni, kardiovaskularni, gastrointestinalni sustav te kožu. Stupanj ozbiljnosti simptoma može varirati od blagih do opasnih po život. Radi se o gubitku oralne podnošljivosti alergena, a to dovodi do senzitizacije i alergije. Najčešći alergeni u dječjoj dobi su bjelančevine kravljeg mlijeka, soja, gluten, orašasti plodovi i jaja, ribe, rakovi te školjke. S vremenom će se na neke alergene razviti tolerancija. Razvoju alergije u djeteta, uz nasljeđe i način života, doprinosi i nedostatak vitamina D, kasno uvođenje dohrane, pretilost te promjene u načinu ishrane. Dijagnoza se postavlja na temelju obiteljske i osobne anamneze, in vivo i in vitro testovima, oralnim provokacijskim testom, gastroenterološkim pretragama te isključnom dijetom. Dugoročno liječenje alergija na hranu se svodi na eliminaciju alergene hrane iz prehrane. Posljednjih desetljeća se provode brojna istraživanja vezana za primjenu imunoterapije u liječenju alergija na hranu, kao i istraživanja za provođenje efikasnih postupaka prevencije senzitizacije djece na nutritivne alergene te time smanjenja rizika za pojavu alergije na hranu u dječjoj dobi. Food allergies are immune reactions of the body and diseases that recur after ingestion of a certain food. They represent a significant public health problem, especially in economically developed countries. They are more common among children than in adults and can affect the respiratory, cardiovascular, gastrointestinal system and skin. The severity of symptoms can range from mild to life-threatening. It is a loss of oral tolerance to allergens, which leads to sensitization and allergies. The most common allergens in childhood are cow's milk proteins, soy, gluten, nuts and eggs, fish, crabs and shellfish. Tolerance to some allergens will develop over time. In addition to heredity and lifestyle, vitamin D deficiency, late introduction of supplementation, obesity and changes in diet contribute to the development of allergies in children. The diagnosis is made on the basis of family and personal history, in vivo and in vitro tests, oral provocation test, gastroenterological tests and food elimination diet. Long-term treatment of food allergies comes down to eliminating food allergens from the diet. In recent decades, numerous studies related to the application of immunotherapy in the treatment of food allergies, as well as research to conduct effective procedures to prevent sensitization of children to nutritional allergens and thus reduce the risk of food allergies in childhood.

Sumpor se u vinarstvu koristi stoljećima, kako je vrijeme pokazalo, zbog svojih antioksidativnih i antiseptičkih svojstava. Zamjena mu još nije pronađena, koja bi udovoljila zahtjevima trajnosti i biološke stabilnosti. Negativni učinci upotrebe sumporovog dioksida uglavnom su vezani za njegovu primjenu u prekomjernim količinama. Visoke koncentracije sumporovog dioksida ugrožavaju miris i okus vina i znatno povećavaju rizik od stvaranja sumporovodika. S obzirom da može izazvati alergijske reakcije, sumporov dioksid je označen kao alergen i njegova koncentracija u vinima je na tržištu zakonski regulirana. Iz svega navedenog, nužno je da se sumporov dioksid kvantificira u vinu pomoću različitih metoda., Sulfur dioxide has been used in winemaking for centuries, as time has shown, for its antioxidant and antiseptic properties. No replacement has been found yet, which would meet the requirements of durability and biological stability. The negative effects of sulfur dioxide use are mainly related to its use in excessive amounts. High concentrations of sulfur dioxide compromise the aroma and taste of wine and significantly increase the risk of hydrogen sulfide formation. Because it can cause allergic reactions, sulfur dioxide is designated as an allergen and its concentration in wines is regulated by law. From all the above it is necessary to quantify sulfur dioxide in wine by different methods.

Cilj: Odrediti profil alergijske senzitizacije i okidače alergijskih simptoma prema dobi djeteta i tipu alergijske bolesti. Ispitanici i metode: Restrospektivno su analizirani rezultati kožnih ubodnih testova na inhalatorne i nutritivne alergene u lokalnoj populaciji ispitanika (n=1102; dob 0-18 god; 58% (n=640) dječaka i 42% (n=462) djevojčice) koji boluju od: astme s pridruženim alergijskim rinitisom ili rinokonjunktivitisom 29% (n=323), alergijskog rinitisa 24% (n=269), atopijskog ekcema 21% (n=226), alergijskog rinokonjunktivitisa 13% (n=148), astme 8% (n=85), te akutne alergijske urtikarije 5% (n=51). Podaci o neposredno zamijećenim okidačima alergijskih simptoma dobiveni su analizom anamnestičkih podataka. Rezultati: U ispitanika dojenačke dobi značajno dominira senzitizacija na nutritivne alergene (P, Aim: To determine sensitization profile and allergic triggers related to allergic disease and the age of the child. Subjects and methods: Results of skin prick tests were retrospectively analyzed to inhalational and nutritive allergens in the local population of subjects (n=1102; age 0-18 years; 58% (n=640) boys and 42% (n=462) girls) who suffer from asthma associated with allergic rhinitis or rhinoconjunctivitis 29% (n=323), allergic rhinitis 24% (n=269), atopic eczema 21% (n=226), allergic rhinoconjunctivitis 13% (n=148), allergic asthma 8% (n=85) and acute allergic urticaria 5% (n=51). Data considering diretcly notable triggers of allergic symptoms were obtained from medical history. Results: Sensitization to nutritive allergens dominates in infants (P=0.001). Sensitization to inhalant allergens has siginificant frequency in children after three years of age Sensitization to mite allergens and hairy pets appeared firstly (P=0.003). Sensitization to pollen becomes significant at school age. In subjects with asthma and allergic rhinitis dominate sensitization to inhalant allergens, mostly on mites (P=0.007), except in allergic rhinoconjunctivitis where sensitization to grass pollen is dominant. Subjects with eczema and acute urticaria are mostly sensitized to nutritive allergens such as eggs, nuts and cow's milk. However, sensitization to mites seems to be also important. Allergens are seldom observed as direct triggers of allergic symptoms. In case they are mostly associated with pollination, with hairy pets and with the consumption of eggs. Conclusion: The results are of practical importance for everyday evaluation of allergic sensitization in local population of patients. They can help in the rational selection of current and relevant allergens for skin prick testing taking care of the child's age and its allergic disease or symptoms.

Cilj. Odrediti vrstu i učestalost alergijske preosjetljivosti na alergene u lokalnoj populaciji djece s alergijskim bolestima. Ispitanici i metode. Izvršili smo retrospektivnu analizu rezultata kožnih ubodnih testova na inhalacijske i nutritivne alergene u populaciji djece Šibensko-kninske županije (n = 704) u dobi od 0 do 18 godina. Od ispitane djece bilo je 57,14% (n = 411) muške djece i 42,85% (n = 293) ženske djece. Od ukupnoga broja ispitanika 37,5% (n = 264) djece imalo je dijagnozu astme, 22,0% (n = 155) virusima induciranih bronho-opstrukcija, 14,4% (n = 102) atopijskog dermatitisa, 20,0% (n = 141) alergijskog rinitisa/rinokonjunktivitisa i 5,9% (n = 42) alergijske urtikarije. Rezultati. U dojenačkoj dobi učestalija je preosjetljivost na nutritivne alergene (P < 0,01), u prvom redu na alergene cijelog kokošjeg jajeta. Od inhalacijskih alergena najprije se pojavljuje preosjetljivost na alergene grinja, dlakavih kućnih ljubimaca i peludi trava. Od treće godine učestalija je preosjetljivost na inhalacijske alergene (P < 0,001). U ispitanika s astmom prevladava preosjetljivost na inhalacijske alergene (P < 0,001). Grinje su značajno češći uzročnici astme u odnosu na ostale inhalacijske alergene (P < 0,001). U ispitanika s izoliranim alergijskim rinitisom/alergijskim rinokonjunktivitisom prednjači preosjetljivost na pelud trava. Zaključak. Ispitivanjem smo odredili učestalost alergijske preosjetljivosti na alergene u populaciji djece Šibensko-kninske županije. Dokazali smo visoku učestalost preosjetljivosti na grinje kućne prašine i alergene peludi trava. Sukladno dobivenim rezultatima prilikom procjenjivanja alergijske preosjetljivosti kod izbora inhalacijskih alargena, predlažemo djecu iznad treće godine testirati na grinje kućne prašine, pelud trava, pelud masline, alergene dlakavih kućnih ljubimaca, ambroziju, brezu, čempres i pelin., Goal. To determine hypersensitivity to allergens in the local child population with allergic diseases. Subjects and methods. We have carried out a retrospective analysis of the skin prick test results for inhalant and nutritional allergens in the local child population (n = 704) aged 0 to 18 years. 57.14% (n = 411) of the examined children were male and 42.85% (n = 293) female. 37.5% (n = 264) of the total number of subjects had a diagnosis of asthma, 22.0% (n = 155) of virus-induced bronchial obstructions, 14.4% (n = 102) atopic dermatitis, 20.0% (n = 141) allergic rhinitis/rhinoconjuctivitis and 5.9% (n = 42) allergic urticaria. Results. In infancy, hypersensitivity to nutritional allergens (P < 0.01) is more frequent, primarily to the allergens of the whole chicken egg. As for the inhalant allergens, hypersensitivity to mite, hairy pets and grass pollen allergens appears first. From the age of three, hypersensitivity to inhalant allergens (P < 0.001) is more frequent. Hypersensitivity to inhalant allergens prevails in subjects with asthma (P < 0.001). Mites are significantly more frequent asthma causes than other inhalant allergens (P < 0.001). Hypersensitivity to grass pollen predominates in subjects with isolated allergic rhinitis/rhinoconjuctivitis.Conclusion. In the study, we have determined the incidence of hypersensitivity to allergens in the child population of Šibenik-Knin County. We have proved a high incidence of hypersensitivity to house dust mites and grass pollen allergens. In accordance to the results obtained, when evaluating allergic hypersensitivity, we suggest that, in the selection of inhalant allergens, children above 3 years of age are tested for house dust mites, grass pollen, olive tree pollen, hairy pet allergens, Ambrosia (ragweed), birch, cypress and wormwood.

Alergije su jedan od najčešćih uzročnika mnogih zdravstvenih problema u ljudskoj populaciji, kako kod odraslih, tako i kod djece. Alergija se tumači kao reakcija organizma na neku njemu nepoznatu i inače neopasnu tvar iz neposredne okoline. Tvari koje izazivaju alergije nazivaju se alergeni i sve ih je više poznatih posljednjih godina. Organizam reagira na nepoznati alergen obrambenim mehanizmom koji stvara histamin, bjelančevinu kojom se bori protiv alergije. Alergije pogađaju jednu četvrtinu hrvatske populacije, dok je u Americi taj broj izražen u milijunima. Alergije na hranu manje su proširene – puno ljudi se žali kako imaju alergiju na hranu, no često se ona zamjenjuje sa intolerancijom na hranu. One mogu izazvati i više od puke nelagode i blažih simptoma poput hunjavice, svrbeža, kašlja i sl. Nekada alergijska reakcija može postati toliko snažna da izazove anafilaktički šok prilikom kojega može doći i do gušenja, kolapsa organizma i naposljetku do smrti. Veoma je važno na vrijeme primijetiti simptome alergije te početi koristiti terapiju koja će ublažiti simptome te olakšati život alergičara. Allergies are one of the most common causative agent most od the health problems in human population, by adults and by children. Allergy is interpreted as a reaction to some unknown and it normally harmless substance from the immediate surroundings. Substances that causes allergyes are called allergens and they are so more and more well known allergens in late years. The body responds to an unknown allergen defensive mechanism that produces histamine, a protein that fights allergies Allergies affects one quarter of Croatian population and in America, that number is expressed in milions. Food allergies are less enlarged - a lot of people complain that they have food allergies, but often it is replaced with a food intolerance They can cause more than just discomfort and gently symptomes like rhinitis, itching and coughing. Sometimes an allergy reaction can became so strong that cause anaphylaxis during which can come to choking, collapse of organism and death. It is very imortant on time notice the allergies and start to use theraphy to alleviate the symptoms of allergy and make life easier for those with allergies.

Cilj: Odrediti profil alergijske senzitizacije i okidače alergijskih simptoma prema dobi djeteta i tipu alergijske bolesti. Ispitanici i metode: Restrospektivno su analizirani rezultati kožnih ubodnih testova na inhalatorne i nutritivne alergene u lokalnoj populaciji ispitanika (n=1102; dob 0-18 god; 58% (n=640) dječaka i 42% (n=462) djevojčice) koji boluju od: astme s pridruženim alergijskim rinitisom ili rinokonjunktivitisom 29% (n=323), alergijskog rinitisa 24% (n=269), atopijskog ekcema 21% (n=226), alergijskog rinokonjunktivitisa 13% (n=148), astme 8% (n=85), te akutne alergijske urtikarije 5% (n=51). Podaci o neposredno zamijećenim okidačima alergijskih simptoma dobiveni su analizom anamnestičkih podataka. Rezultati: U ispitanika dojenačke dobi značajno dominira senzitizacija na nutritivne alergene (P, Aim: To determine sensitization profile and allergic triggers related to allergic disease and the age of the child. Subjects and methods: Results of skin prick tests were retrospectively analyzed to inhalational and nutritive allergens in the local population of subjects (n=1102; age 0-18 years; 58% (n=640) boys and 42% (n=462) girls) who suffer from asthma associated with allergic rhinitis or rhinoconjunctivitis 29% (n=323), allergic rhinitis 24% (n=269), atopic eczema 21% (n=226), allergic rhinoconjunctivitis 13% (n=148), allergic asthma 8% (n=85) and acute allergic urticaria 5% (n=51). Data considering diretcly notable triggers of allergic symptoms were obtained from medical history. Results: Sensitization to nutritive allergens dominates in infants (P=0.001). Sensitization to inhalant allergens has siginificant frequency in children after three years of age Sensitization to mite allergens and hairy pets appeared firstly (P=0.003). Sensitization to pollen becomes significant at school age. In subjects with asthma and allergic rhinitis dominate sensitization to inhalant allergens, mostly on mites (P=0.007), except in allergic rhinoconjunctivitis where sensitization to grass pollen is dominant. Subjects with eczema and acute urticaria are mostly sensitized to nutritive allergens such as eggs, nuts and cow's milk. However, sensitization to mites seems to be also important. Allergens are seldom observed as direct triggers of allergic symptoms. In case they are mostly associated with pollination, with hairy pets and with the consumption of eggs. Conclusion: The results are of practical importance for everyday evaluation of allergic sensitization in local population of patients. They can help in the rational selection of current and relevant allergens for skin prick testing taking care of the child's age and its allergic disease or symptoms.

Nutritivna alergija je klinička rekacija na bjelančevine hrane. Za nastanak odgovoran je prvi tip preosjetljivosti posredovan IgE protutijelima. Najčešći alergeni su mlijeko, soja, ţitarice, riba, školjke, jaja, kikiriki i orašasti plodovi. Alergije zahvaćaju ljude iz cijeloga svijeta, no nejednakim intezitetom. U dječjoj dobi najučestalije su kod dojenčadi. Prevalencija se kreće 3 % - 10 %. Očituje se urtikarijom, angioedemom, eritemom uz svrbeţ koţe. Često dolazi do gastrointestinalnih simptoma, rijetko do respiratornih simptomima. Moguća je pojava generalizirane anafilaktičke reakcije. Dijagnoza se zasniva na anamnezi, fizikalnom pregledu, kliničkoj slici, koţnim testovima i laboratorijskoj dijagnostici. Liječenje nutritivnih alergija se sastoji od prekidanja izlaganja uzročnom alergenu, protuupalnog i simptomatskog liječnja. Sestrinska skrb usmjerena je na prevenciju, edukaciju te na zbrinjavanje djece i njihovih obitelji. Food allergy is a clinical reaction to food proteins. Responsible for its occurrence is the first type of hypersensitivity mediated by IgE antibodies. The most common allergens are milk, soy, wheat, fish, shellfish, eggs, peanuts and tree nuts. Allergies affect people from all over the world, but not in the equal intensity. In pediatric patients, infants are most likely to have a food allergy. Prevalence ranges from 3 % to 10 %. They manifest with urticaria, angioedema, erythema with itching skin.. It often leads to gastrointestinal symptoms, rarely to respiratory symptoms. There is a possibility of general anaphylactic reaction. Diagnosis is based on medical history, physical exam, clinical manifestations, skin tests and laboratory tests. Treatment of food allergies consists of avoidance of causing allergen, anti - inflammatory and symptomatic treatment. Nursing care is focused on prevention, education and care of children and their families.

Summary The purpose of this study was to analyze the relation between clinical picture of allergic disease and the level of pollen count the patients are exposed to. Material and method We analyzed the severity of allergic rhinitis, conjunctivitis and bronchial asthma in a group of 640 subjects from Warsaw area with hypersensitivity to hazel, alder, grass, mugwort, plantain, nettle, Alternaria and Cladosporium allergens. Aerobiological measurements, patients’ symptoms score cards and the results of physical examination were analyzed. Results First symptoms in patients allergic to grass were visible during exposure to the concentration of 20 pollen grains in 1 m3 of air (25% subjects sensitised to grass pollen). Symptoms were noted in all the subjects sensitized to grass pollen at a concentration of approximately 50 grains/m3 of air. During exposure to the concentration of 65 pollen grains per m3 the symptoms were strengthened. Several hours’ exposure to grass pollen concentration exceeding 120 grains/m3 cause dyspnoea in some patients. Similar intensive symptoms occurred after exposure to alder, birch and mugwort pollen. Patients with positive skin prick tests to plantain and nettle pollen allergens experienced symptoms of low intensity or even none symptoms during pollination period. Subjects with hypersensitivity to Alternaria allergens experienced symptoms during exposure to the concentration of approximately 80 spores in 1 m3 of air, while patients sensitised to Cladosporium allergens, during exposure to the concentration of over 2800 spores in 1 m3 of air. The dominant symptom occurring in patients sensitized to fungal allergens was loss of nasal patency, and dyspnoea as well as recurring cough at higher concentrations. Conclusions Clinical symptoms of allergic disease were connected to the concentration and the kind of aeroallergen the subjects were exposed to.

Prehranska alergija je imunska reakcija na določeno živilo, ki lahko pri posamezniku izzove različne simptome, tudi smrt, zato je toliko bolj pomembno, da se posameznika zaščiti pred takšnimi reakcijami. V nalogi so obravnavane uredbe, direktive ter slovenski pravilnik s področja označevanja živil ter varnosti živil, s poudarkom na področju označevanja alergenov. S pomočjo anketnega vprašalnika objavljenega na spletu med februarjem in aprilom 2013, z vprašanji vezanimi na doživljanje potrošnikov na prehranske alergije, sem pridobila podatke, ki sem jih ovrednotila s programom SPSS. Rezultatov razširjenosti prehranskih alergij v Sloveniji ne moremo primerjati s splošno znanimi podatki, so se pa kot najpogostejše v naši anketni raziskavi pokazale alergije na mleko in mlečne izdelke, gluten, jajca ter oreške in arašide. Težave zaradi prehranskih alergij pri večini anketirancev vplivajo na kvaliteto življenja predvsem zaradi povečane skrbi glede označb na živilih, povečanega finančnega izdatka zaradi izbire primernih živil, strahu pred zaužitjem škodljivega živila ter omejenega prehranjevanja zunaj doma. Anketiranci so bili tudi večinskega mnenja, da živilski izdelki, ki vsebujejo alergene sestavine niso dovolj dobro označeni in da zakonodaja ne skrbi dovolj dobro za posameznike s prehranskimi alergijami. Obravnavana živilska zakonodaja se prilagaja potrebam posameznikov z alergijami vendar pa so spremembe še mogoče in tudi s strani dovzetnih posameznikov pričakovane. Food allergy is an immune reaction to a certain food that can cause a variety of symptoms, even death. Therefore it is all the more important to protect the individual against such reactions. In my thesis I will discuss the regulations, directives and Slovenian rules on food labelling and food safety, with an emphasis on the labelling of allergens. With the help of a questionnaire published on the Internet between February and April 2013, which included questions related to the perception of consumers to food allergies, I have gained valuable information that I have evaluated with SPSS. We cannot compare the prevalence of food allergies in Slovenia with the generally available information on the prevalence of food allergies. The results of our questionnaire showed that the most commonly manifested food allergies are allergy to milk and milk products, gluten, eggs, nuts and peanuts. Problems due to food allergies mostly affect the quality of life. This is mainly because people have to look through food labels on a daily basis, which increases their concerns. In addition people have increased financial expenses due to the selection of appropriate foods. Moreover, they are also limited when it comes to eating out and also fear to eat harmful foods outside their home. Respondents in majority thought that food products containing allergenic ingredients are not well marked and that the law does not care enough for individuals with food allergies. The present food law is trying to adapt to the needs of individuals with allergies but there is still, room for changes, which open-minded individuals also expect.

The subject matter of the bachelor thesis is specific allergen immunotherapy from the perspective of nurses. The work is divided into theoretical and practical parts. The concepts associated with the topic are explained in the theoretical section of the thesis. These concepts are as follows: allergy, allergen, immunity, sensitization, anaphylaxis and specific allergen therapy. Basic allergic diseases, work and role of the nurse are also described in the theoretical part. The practical part of the thesis contains four goals: the first three goals relate to nurses working in allergology ambulances; the fourth goal is focused on the needs of patients undergoing treatment with specific allergen immunotherapy. The first goal was to find out nurses' knowledge related to the specific allergen immunotherapy, second one dealt with the skills of nurses in the administration of specific allergen immunotherapy and the patients education process realised by nurses in allergology ambulances represented the third goal of the thesis. The fourth objective focused on the needs of patients undergoing treatment with specific allergen immunotherapy. To achieve these objectives the qualitative inquiry method of semi-structured interviews and observations were used. The research group consisted of four nurses working in private allergology ambulances in South Bohemia and four patients undergoing treatment with specific allergen immunotherapy. The research showed that nurses working in allergology ambulances have enough knowledge and skills in administration of specific allergen immunotherapy. The primary needs of patients can be determined as the need to be informed and to perceive the positive attitude of nurses and physicians to the patients.

Ph.D. THESIS TITLE Nutritional Biology of Synanthropic Mites (Acari: Acaridida) ABSTRACT Several attempts to describe the nutritional biology of acaridid mites were undertaken, however full understanding of these processes remains incomplete. The objective of this Ph.D. thesis was to expand our knowledge concerning digestive physiology of stored product and house dust mites and to apply this knowledge to their nutritional biology. The research approach adopted in this Ph.D. thesis includes in vitro characterization of enzymatic activity in whole mite extracts (WME) and spent growth medium extracts (SGME), evaluation of the enzyme activities with respect to the gut physiological pH, enzyme inhibition experiments, in vivo localization of enzyme activities in the mite gut, determination of effects of nutrient or antifeedant additives in experimental diets on mite population growth and determination of the feeding preferences of synanthropic mites as assessed by in vitro and in vivo analyses. The gut contents of twelve species of synanthropic acaridid mites were determined to be within a pH range of 4 to 7 and showed a pH gradient from the anterior to the posterior midgut. The pH in digestive tract of synanthropic acaridid mites corresponds to the activity of proteases, α-glucosidases, α-amylases and...

This diploma thesis was focused on the study of lysozyme and lysozyme-like proteins, either of similar function (antibacterial) or molecular weight (14 - 17 kDa), of synanthropic acaroid mites. In general, animals utilize lysozymes for defensive (antimicrobial) or digestive purposes but also as a digestive enzyme. Some chitinases or other enzymes that act similarly to lysozyme can be utilized for similar purposes. Chitinases belong to house dust mite allergens. One of major mite are historically named lysozyme-like proteins which name relates to their size similar to lysozyme. Bacteriolytic activity has also 14.5 kDa (UniprotKB Q8MWR6) protein. The species selected for the study were domestic mites Dermatophagoides farinae, D. pteronyssinus and Lepidoglyphus destructor. Presence of lysozyme was detected by direct detection with polyclonal antibody using immunohistochemistry and dot blots. Immunohistochemistry proved presence of lysozyme epitopes in the feces of D. farinae, D pteronyssinus a L. destructor. Dot blot analysis demonstrated the presence of imunoreactivity of antibody in spent growth medium extracts (SGME) of all three species. This implies that lysozyme is synthesized in the midgut. The presence of lysozyme and lysozyme-like proteins was proved using 2D electrophoresis and MALDI TOF/TOF...

As the subject of my Bachelor{\crq}s work I choose "Allergic diseases in relation to risk factors of the life style of children of younger school age in the region Karlovy Vary" because I think that allergies represent a topical problem which due to its nature affects not only children but also our whole society. The aim of the work is to find out how the selected life style factors share in the occurrence of allergies of children of younger school age in the region Karlovy Vary. The work comprises a theoretical and a practical part. The theoretical part comprises comprehensive theoretical knowledge from the field of allergology that should outline the conditions for the development of allergies, the occurrence of allergies in course of the life, the most common allergic diseases and their subsequent diagnosis and treatment. The practical part is devoted to the realised research which was carried out from November 2009 to January 2010 in regional towns and municipalities of the region Karlovy Vary. The research was performed using the form of the quantitative research, the inquiry method and as the data collection technique was used the anonymous questionnaire. The research file included children of younger school age from 4th and 5l classes of primary schools. As the results of the research show, the children who live in big cities have almost comparable occurrence of allergies as the children from small villages. The results also show that the children who are the most exposed to a smoky environment at home have also the biggest occurrence of allergies in contrast to the children who are not exposed to a tobacco environment at all and for whom predominate an absence of allergies. The investigation into whether the lack of microbes influences the occurrence of children allergies demonstrated in case of using disinfecting agents that shorter periods of their usage have an influence on the occurrence of allergies and to the contrary. The research also shows that elements which share in the character of housing have in case of allergies quite individual displays and therefore can not be denominated as significant factors in general. The results of this research can bring useful information to children allergists as well as wide public when the acquired knowledge can increase the interest in these problems and therefore improve preventive and repressive measures in this field.

Praca przedstawia przebieg sezonu pylenia topoli w wybranych miastach Polski w 2008 roku. Pomiary wykonano w Białymstoku, Bydgoszczy, Drawsku Pomorskim, Krakowie, Lublinie, Olsztynie, Szczecinie i Warszawie. Badania prowadzono metodą objętościową przy wykorzystaniu aparatów firmy Burkard i Lanzoni 2000. Sezon pyłkowy wyznaczono jako okres, w którym w powietrzu występuje 95% rocznej sumy ziaren pyłku. Początek sezonu pyłkowego wyznaczono również metodą 30 ziaren. Indeks SPI obliczono jako sumę średnich dobowych stężeń pyłku w danym sezonie. Najwyższe wartości średniodobowych stężeń pyłku topoli odnotowano w Lublinie, gdzie 11 kwietnia stężenie pyłku wyniosło 273 z/m3 powietrza. This paper presents the course of poplar pollination season in selected cities of Poland in 2007. The measurements were performed in Bialystok, Bydgoszcz, Krakow, Lublin, Lody, Olsztyn, Sosnowiec, Szczecin, Warsaw and Wroclaw. Volumetric method with the use of Volumetric Spore Trap (Burkard, Lanzoni 2000) was implemented. Pollen season was defined as the period in which 95% of the annual total catch occurred. The start of the season was also determined by the 30 grains method. Seasonal Pollen Index (SPI) was estimated as the annual sum of daily average pollen concentrations. The highest 24-hour average pollen count was recorded in Lublin on 11 April (273 poplar pollen grains/1 m3).

Praca przedstawia przebieg sezonu pylenia dębu w wybranych miastach Polski w 2008 r. Pomiary wykonano w Białymstoku, Bydgoszczy, Krakowie, Lublinie, Łodzi, Olsztynie, Sosnowcu, Szczecinie, Warszawie i we Wrocławiu. Badania prowadzono metodą objętościową przy wykorzystaniu aparatów firmy Burkard i Lanzoni 2000. Sezon pyłkowy wyznaczono jako okres, w którym w powietrzu wystąpiło 95% rocznej sumy ziaren pyłku. Początek sezonu pyłkowego wyznaczono również metodą 30 ziaren. Indeks SPI obliczono jako sumę średnich dobowych stężeń pyłku w danym sezonie. Najwcześniej sezon pylenia dębu rozpoczął się we Wrocławiu (25 kwietnia). Najwyższe wartości średniodobowych stężeń pyłku dębu odnotowano w Lublinie, gdzie 5 maja stężenie wyniosło 212 z/m3 powietrza. This paper presents the course of oak pollination season in selected cities of Poland in 2007. The measurements were performed in Bialystok, Bydgoszcz, Krakow, Lublin, Lodz, Olsztyn, Sosnowiec, Szczecin, Warsaw and Wroclaw. Volumetric method with the use of Volumetric Spore Trap (Burkard, Lanzoni 2000) was implemented. Pollen season was defined as the period in which 95% of the annual total catch occurred. The start of the season was also determined by the 30 grains method. Seasonal Pollen Index (SPI) was estimated as the annual sum of daily average pollen concentrations. Pollen season of oak in 2008 started first in Lublin (25 April). The highest 24-hour average pollen count was recorded in Lublin on 5 May (212 oak pollen grains/m3).

Praca przedstawia przebieg sezonu pylenia cisu/jałowca w wybranych miastach Polski w 2007 r. Pomiary wykonywano w Białymstoku, Bydgoszczy, Lublinie, Sosnowcu, Szczecinie, Warszawie i we Wrocławiu. Badania prowadzono metodą objętościową przy wykorzystaniu aparatów firmy Burkard i Lanzoni 2000. Za sezon pyłkowy uznano okres, w którym w powietrzu występuje 95% rocznej sumy ziaren pyłku cisu/jałowca. Najwcześniej sezon pylenia cisu rozpoczął się we Wrocławiu, bo już 1 marca. Najpóźniej pyłek cisu pojawił się w Białymstoku – 9 marca. Najwyższe wartości średniodobowych stężeń pyłku cisu odnotowano w Szczecinie, gdzie 13 marca zanotowano stężenie 239 ziaren pyłku w 1m3 powietrza. his paper presents the course of yew/juniper pollination season in selected cities of Poland in 2007. The measurements were performed in Bialystok, Bydgoszcz, Lublin, Sosnowiec, Szczecin, Warsaw and Wroclaw. Volumetric method with the use of Volumetric Spore Trap (Burkard, Lanzoni 2000) was implemented. Pollen season was defined as the period in which 95% of the annual total catch occurred. Pollen season of yew /juniper in 2007 started first in Wroclaw (01 March). The highest 24-hour average pollen count was recorded in Szczcin on 13 March (239 pollen grains/1m3).

Praca przedstawia przebieg sezonu pylenia traw w roku 2007 w Białymstoku, Bydgoszczy, Krakowie, Lublinie, Łodzi, Rzeszowie, Sosnowcu, Szczecinie, Warszawie i we Wrocławiu. Badania prowadzono metodą objętościową przy zastosowaniu aparatów typu Burkard i Lanzoni. Sezon pyłkowy wyznaczono metodą 98%. Najwcześniej pyłek traw zarejestrowano we Wrocławiu i w Krakowie (20, 30 kwietnia), najpóźniej w Białymstoku (13 maja). Największą maksymalną koncentrację pyłku zanotowano w Łodzi (513 ziaren/m3), najniższą w Krakowie (106 ziaren/m3). Okres maksymalnych stężeń pyłku traw rejestrowano między 29 maja i 21 czerwca. This paper presents the curse of grass pollination seasons in the cities Białystok, Bydgoszcz, Kraków, Lublin, Łódź, Rzeszów, Sosnowiec, Szczecin, Warszawa and Wrocław in 2007. Measurements were performed by the volumetric method with the use of a Burkard and Lanzoni Spore Trap. The duration of the pollen seasons was defined as the period in which 98 % of the annual total catch occurred. The pollen seasons of grass started first in Wrocław and Kraków (20, 30 April), the latest in Białystok (13 May). The highest concentration of pollen grains was noted in Łódź (513 z/m3), the lowest in Kraków (106 z/m3). The period of the maximum concentrations was recorded in 29 May and the off 21 June.

Nickel is the most common substance in our environment. It is known as commonest couse of allergic contact dermatitis. Similar to chromium, cobalt and zinc, nickel may also be a cause of allergy in the upper and lower airways. It may lead to diseases described as "hard metal asthma" and "hard metal rhinitis". The study aimed to determine the prevelance of systemic nickel induced reaction in patients sensitized to nickel and suffering from atopic dermatitis, allergic contact dermatitis and urticaria. In all patients during remission of symptoms were performed patch tests with European Standard contact allergens, serial dilution patch tests and oral provocation test with increasing doses of nickel sulphate. Our studies indicate that oral nickel challenge can provoke symptoms of skin allergy type I and type IV.

Pollens from Ambrosia species have a strong allergenic potential and they represent a complex mixture of at least 52 different antigens. This weed is widespread in Serbia and has a long pollination period (appr. 100 days). Pollens are present in very high concentrations during the pollination season (up to 300 pg/m3 of air). Fifty dogs with a clinical diagnosis of canine atopic dermatitis (CAD) and positive intradermal skin test (IDST) reactions to other environmental allergens were included in this study along with 30 dogs without clinical signs of CAD. Both groups were tested intradermally with Ambrosia artemisiifolia extract in concentration of 1000 PNU/mL (provided by ''Institute of virology, vaccines and sera - Torlak''). We have found that Ambrosia extract was second most common allergen to yield positive intradermal test reactions just after house dust mites. The positive reaction of IDST was observed in 66% of the atopic dogs and in 10% of clinically healthy dogs. Dogs that had a positive reaction on intradermal skin test to Ambrosia pollens, mostly showed secondary skin lesions on the pads and perioral skin. In 88% of cases, Malassezia pachydermatis have been isolated from the skin in an increased number and 64% of dogs had otitis externa. These results indicate, that allergens from Ambrosia artemisifolia are of great importance in the etiopathogenesis of atopic dermatitis in dogs originating from the Belgrade region. Polen ambrozije ima jaka alergijska svojstva i sastoji se od kompleksne mešavine od do sada poznatih 52 antigena. Ova korovska biljka je znatno rasprostranjena u Srbiji i ima dugačak period polinacije od prosečno 100 dana. Polen je prisutan u izrazito visokim koncentracijama tokom avgusta i septembra ide i do 300 polenovih zrna u m3 vazduha. Ovim ispitivanjem obuhvaćeno je 50 pasa kod kojih je dijagnostikovan atopijski dermatitis na osnovu kliničkih simptoma i pozitivne reakcije na intradermalnom testu, kao i 30 pasa bez klinički manifestnih simptoma atopijskog dermatitisa. Kod obe grupe pasa uradjeno je intradermalno testiranje standardnim rastvorima proizvođača Instituta za imunologiju i virusologiju Torlak, kao i vodenim rastvorom alergena ambrozije u koncentraciji 1000 PNU/ml. U našem ispitivanju polen ambrozije predstavlja drugi alergen po učestalosti pozitivne reakcije na intradermalnom testu kod ispitivanih pasa, odmah posle kućnih grinja. Pozitivna reakcija zabeležena je kod 66% pasa iz grupe atopičara i kod 10% klinički zdravih pasa. Kod pasa koji su pokazali pozitivnu reakciju na intradermalnom testu na polen ambrozije, sekundarne lezije kože su se najčešće javljale na šapama i oko usana. Kod 88% ovih pasa na koži je u povećanom broju dijagnostikovana Malassezia pachydermatis, a kod 64% jedinki je registrovan otitis externa različite etiologije. Na osnovu dobijenih rezultata se može zaključiti da je alergen biljke Ambrosia artemisiifolia veoma značajan u indukciji atopijskog dermatitisa kod populacije pasa sa šireg područja Beograda.

Art v 1 is the major allergen of mugwort (Artemisia vulgaris) pollen, a significant cause of hay fever all over Europe. Specific immunotherapy is the only treatment modality for allergic disease. Application of modified allergens makes the treatment safer and more efficient. In this work, two out of three (citraconic anhydride, cis-aconitic anhydride, 2,3-dimethylmaleic anhydride) tested anhydrides were proven to be suitable for chemical modifications of allergens. Art v 1 was modified by cis-aconitylation and citraconylation in order to obtain derivatives of Art v 1 that may be suitable for further immunological testing. Acylation of Art v 1 gave derivatives (caaArt v 1 and citArt v 1) with about 80 % modified amino groups. The derivatives were in the monomeric form and had dramatically reduced pI values. Both derivatives were relatively stable at neutral pH values, while the acyl groups undergo hydrolysis under acidic conditions. Modification of allergens by cis-aconitylation and citraconylation could be a new tool for obtaining allergoids. Art v1 je glavni alergen polena crnog pelina (Artemisia vulgaris), značajnog uzročnika polenske groznice širom Evrope. Alergen-specifična imunoterapija je za sada jedini delotvoran način za tretiranje alergija, pri čemu primena modifikovanih alergena čini ovakav tretman bezbednijim i efikasnijim. U ovom radu, dva od tri (anhidrid cis-akonitne, citrakonske i 2,3-dimetilmaleinske kiseline) ispitivana anhidrida su se pokazala pogodnim za hemijske modifikacije alergena. Art v 1 je modifikovan cis-akonitilovanjem i citrakonilovanjem u cilju dobijanja derivata Art v 1 pogodnih za dalje imunološke testove. Acilovanjem Art v 1 dobijeni su derivati (caaArt v 1 i citArt v 1) sa oko 80 % izmodifikovanih amino grupa. Dobijeni derivati su monomerni, sa molekulskom masom sličnom nativnom Art v 1, ali sa dramatično smanjenim pI vrednostima. Oba derivata su relativno stabilna u neutralnoj, dok se u kiseloj sredini acil grupe hidrolizuju. Modifikacija alergena cis-akonitilovanjem i citrakonilovanjem može biti novi način za dobijanje alergoida.

House dust mites (HDM) represent a major source of allergens, contributing to the increasing incidence of type I hypersensitivity disease worldwide. Over 30 different IgE-binding proteins from the HDM extract were detected. Although group 1 and 2 have been identified as major allergens, due to the safety and efficacy of allergy diagnosis and immunotherapy, there is a need to carefully evaluate the clinical relevance of other allergens present in the HDM extract. In regard to this, a high molecular mass allergen of about 68 kD was purified from the HDM extract using a combination of gel permeation chromatography and reversed-phase chromatography. The IgG and IgE reactivity of the purified protein were preserved during the purification process, as confirmed by Western blot analysis with polyclonal rabbit antibodies and dot blot analysis with a pool of sera from subjects with house dust mite allergy, respectively. In addition, the IgE reactivity was confirmed using ELISA testing with nine patient sera. The biological potency of the 68 kD allergen was confirmed by skin prick testing in five allergic subjects, suggesting that the high molecular mass allergen is a good candidate for component-resolved diagnosis of house dust mite allergy and eventual therapeutic treatment. Grinje iz kućne prašine predstavljaju jedan od glavnih izvora alergena koji su u značajnoj meri doprineli porastu prvog tipa preosetljivosti. Preko 30 IgE-vezujućih proteina iz kućne prašine je detektovano do danas. Alergeni grupe 1 i 2 označeni su kao glavni alergeni kućne prašine. Međutim, da bi se poboljšala sigurnost i efikasnost dijagnoze i terapije alergijskih oboljenja izazvanih grinjama iz kućne prašine, neophodno je odrediti klinički značaj svih alergena iz ovog alergenskog izvora. U ovom radu izolovan je alergen visoke molekulske mase od 68 kD iz ekstrakta kućne prašine kombinovanjem gel-permeacione hromatografije i reversno-fazne hromatografije. IgG i IgE reaktivnost prečišćenog proteina je proverena u 'Western blot'-u i 'dot blot'-u sa poliklonskim zečijim antitelima na ekstrakt kućne prašine i 'pool'-om seruma osoba alergičnih na kućnu prašinu, redom. 64 % pacijenata je pokazalo IgE reaktivnost na prečišćeni protein u ELISA testu. Biološka reaktivnost prečišćenog alergena je potvrđena u kožnim probama na pet pacijenata, ukazujući da je prečišćen alergen dobar kandidat za dijagnozu alergije na kućnu prašinu pojedinačnim komponentama i eventualni terapeutski tretman.

Common ragweed (Ambrosia atremisiifolia) is one of the most frequent causes of pollen-induced allergic reactions both in humans and dogs. It has not been defined yet, what is the major allergen(s) to which most dogs allergic to ragweed show a positive result on intradermal skin test (IDST). In the present study sensitization to Ambrosia artemisiifolia pollen allergens in dogs with atopic dermatitis was examined with both in vivo and in vitro tests, including IDST and serum allergen specific IgE test. Detection of specific-IgE antibodies against ragweed allergens by immunoblotting in the sera of allergic dogs was optimized, as well. Dogs that were positive, as judged by IDST reactions to ragweed pollen allergens, also had alergen specific IgE antibodies in their sera. Results indicate that major allergens of A. artemisifolia pollen in dogs are Amb a 1 and Amb a 2. Further characterization of ragweed allergens is needed before they could potentially be used in intradermal testing or allergen immunotherapy in affected dogs. Also, we evaluated new Favrots diagnostic criteria for canine atopic dermatitis in dogs allergic to Ambrosia atremisiifolia pollen. It might be concluded that proposed criteria are of great assistance for seting up suspected diagnosis of canine atopic dermatitis, after ruling out other pruritic dermatoses. Kratka ambrozija (Ambrosia artemisiifolia) je jedan od najčešćih uzročnika alergijskih reakcija izazvanih polenom kod ljudi i pasa. Još uvek nije definisano koji je glavni alergen (i), na koji, većina pasa alergičnih na polen ambrozije, ispoljava pozitivnu reakciju na intradermalnom testu. U ovoj studiji je ispitana senzibilizacija na polen ove biljke kod pasa sa simptomima atopijskog dermatitisa in vivo i in vitro testovima, uključujući intradermalni test i dokazivanje prisustva alergen specifičnih IgE antitela u serumu. Optimizovani su uslovi za detekciju IgE specifičnih antitela iz seruma pasa alergičnih na polen ambrozije imunoblot tehnikom. Psi koji su imali pozitivnu reakciju na polen ove biljke na intradermalnom testu, takođe su imali specifična IgE antitela u serumu. Dobijeni rezultati ukazuju da su glavni alergeni Ambrosia artemisiifolia kod pasa Amb a 1 i Amb a 2. Neophodna je dalja karakterizacija alergena ambrozije kako bi se oni mogli primeniti pri rutinskom intradermalnom testiranju ili u alergen specifičnoj imunoterapiji obolelih pasa. Takođe je razmatrana i validnost Favrotovih dijagnostičkih kriterijuma kod pasa alergičnih na polen ambrozije. Može se zaključiti da su predloženi kriterijumi od velike pomoći u postavljanju suspektne dijagnoze atopijskog dermatitisa pasa, nakon isključenja drugih pruritičnih dermatoza.

Allergic condition such as itching of the skin, or runny nose and sneezing are widely known, but different case of allergic reaction in the mouth. Are still unknown, one of them is Oral Allergy Syndrome (OAS). Almost everyone knows food allergens such as shrimp, or marine fish, but actually fresh fruits and vegetables also can trigger allergic reactions like OAS in the form of itching or swelling of the lips, tongue, palate, and pharynx. Author intends to provide further information about the OAS in this paper. The mechanism of OAS is a type I, immunoglobulin E-mediated hypersensitivity reaction, common in atopic people with history of atopic rhinitis, bronchial asthma, or urticaria. Food allergen that causes OAS have a cross reaction/ homolog with pollen from protein pathogen respons / PR-10 family, such as Bet v 1 (Birch) and Bet v 2 (Birch). Some OAS allergens from vegetables that homolog with Bet v 1 includes celery (Api g 1) and soybean (Gly m 4), and other allergens from fruits homolog with Bet v 2, such as Ana c1 alergen pineapple. The diagnosis of OAS based on medical history, clinical examination and diagnostic tests includes skin prick test, specific IgE serum, or basophil activation test, and cellular antigen stimulation test with an enzyme-linked immunosorbent assay. Management of OAS consists of non-pharmacological includes information and education to avoid the causes, and give suggestion to eat cooked food. Pharmacologically are prescription of antihistamines or epinephrine injection in case of emergencies. Signs and symptoms of OAS in fact have long been found, but its diagnosis and management are still not widely known. Dentist should know about signs and symptoms of food allergy in the oral mucosa include OAS which caused by fruits and vegetables such as celery, soybean, carrot, apple, pineapple, and strawberry, so finally dentist will provide good management for the patient..

beta-Lactoglobulin (BLG) is an important nutrient of dairy products and an important allergen in cow's milk allergy. The aim of this study was to investigate the potential of laccase to cross-link BLG in the presence of an apple phenolic extract (APE) and to characterize the obtained products for their digestibility by pepsin and pancreatin. The composition of the apple phenolics used for cross-linking was determined by liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS). The apple phenolic extract contained significant amounts of quercetin glycosides, catechins and chlorogenic acid. The laccase cross-linked BLG in the presence of apple phenolics. The polymerization rendered the protein insoluble in the reaction mixture. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the cross-linking reaction mixture revealed a heterogeneous mixture of high molecular masses (cross-linked BLG), with a fraction of the BLG remaining monomeric. Enzymatic processing of BLG by laccase and apple polyphenols as mediators can decrease the biphasal pepsin pancreatin digestibility of the monomeric and cross-linked protein, thus decreasing its nutritional value. In addition, reduced BLG digestibility can decrease its allergenic potential. Apple polyphenols can find usage in the creation of new, more functional food products, designed to prevent obesity and hypersensitivity-related disorders. β-Laktoglobulin (BLG) je važan nutrijent mlečnih proizvoda i važan alergen kod alergija na kravlje mleko. Cilj ove studije je bilo ispitivanje potencijala lakaze da unakrsno poveže BLG u prisustvu fenolnog ekstrakta jabuke (APE), kao i karakterizacija dobijenih proizvoda sa aspekta njihove digestibilnosti pepsinom i pankreatinom. Kompozicija fenola jabuke korišćenih za unakrsno povezivanje određena je pomoću LC-ESI-MS. Fenolni ekstrakt jabuke sadrži znatne količine glikozida kvercetina, katehine i hlorogensku kiselinu. BLG je unakrsno povezan lakazom u prisustvu fenola jabuke, pri čemu je polimerizacija učinila BLG nerastvornim u reakcionoj smeši. SDS-PAGE analiza pokazala je da reakciona smeša sadrži heterogenu smešu velikih molekulskih masa (unakrsno povezan BLG), kao i deo zaostalog monomernog BLG. Enzimsko procesovanje BLG lakazom, u prisustvu polifenola jabuke kao medijatora, može smanjiti bifaznu pepsin-pankreatinsku digestibilnost kako monomernog, tako i unakrsno povezanog BLG, i na taj način smanjiti njegovu nutritivnu vrednost. Takođe, smanjena digestibilnost BLG može smanjiti njegov alergeni potencijal. Polifenoli jabuke mogu se koristiti za kreiranje novih, funkcionalnijih prehrambenih proizvoda, dizajniranih za prevenciju kako gojaznosti, tako i bolesti vezanih za preosetljivost.

Zamena alergenih ekstrakata pojedinačnim prirodnim i rekombinantnim alergenima u dijagnostici alergije je obećavajući koncept i naziva se dijagnostika razdvojena na komponente CRD (engl. component-resolved diagnosis). U tom smislu, neophodno je proceniti alergeni potencijal svakog pojedinačnog kandidata za primenu u CRD. U ovoj tezi ocenjivane su biohemijske i alergene osobine cistein proteaznog inhibitora (nCPI) iz ploda zelenog kivija. Prečišćeni nCPI pokazao je IgE reaktivnosti in vitro (dot blot) i in vivo (kožne probe i aktivacija bazofila), dok je IgG reaktivnost pokazana u Western blotu i ELISA-i. Biološka aktivnost je ocenjena ispitivanjem antimikrobnog potencijala nCPI naspram tri soja fitopatogenih bakterija (A. tumefaciens, B. cepacia i u manjem obimu bakterije E. carotovora) kao i dva izolata fitopatogenih gljiva (A. radicina i B.cinerea), kako in vitro, tako i in vivo. Zbog slabe zastupljenosti nCPI u plodu kivija kodirajuća sekvenca nCPI je efikasno eksprimirana u Escherichia coli. Molekulska identičnost rCPI u odnosu na prirodni molekul pokazana je strukturnim (maseni otisak prsta) i imunohemijskim (imunoblot i ELISA) i biološkim (inhibicija proteazne aktivnosti papaina i rasta fitopatogenih gljiva) testovima. Na osnovu dobijenih rezultata jasno je da je CPI klinički relevantan alergen i kao takav trebalo bi da bude uključen u dijagnostiku alergije na kivi razdvojenu na komponente. Standardni imunoterapijski pristup u tretmanu alergije na hranu nije adekvatan zbog visokog rizika, pa je primena hipoalergenih molekula koji smanjeno vezuju IgE obećavajući koncept. Diverzifikacija gena za aktinidin rađena je lančanom reakcijom polimeraze sklone greškama epPCR (engl. error-prone PCR) koja je praćena kloniranjem biblioteke u Aga2 ekspresioni sistem za ekspresiju na površini ćelija kvasca. Nakon tri runde sortiranja, nasumično odabrani klonovi su dalje analizirani dot blotom i ELISA testom koristeći serum 6 osoba alergičnih na kivi U poređenju sa prirodnim i rekombinantnim aktinidinom, dva mutanata su pokazala smanjenu sposobnost aktivacije bazofila... The replacement of fruit allergen extracts with component-resolved allergens seems to be a promising concept. In this regard, the evaluation of allergenic properties of a particular allergen candidate for component-resolved diagnosis needs to be performed. In this study evaluation of the biochemical and allergenic properties of cysteine proteinase inhibitor purified from green kiwifruit (nCPI) was performed. The IgE reactivity was detected both in vitro (dot blot) and in vivo (skin prick test and basophile activation assay), while IgG rectivity in both Western blot analysis nd ELISA test. Biological activity was assesed by examining the growth inhibition of three plant pathogenic bacterial strains (A.tumefaciens, B. cepacia and, to a lesser extent, E. carotovora) and two phytopathogenic fungi (A. radicina and B. cinerea) both in vitro and in vivo. Due to low abundance of nCPI molecule in kiwifruit the coding sequence of kiwi CPI was efficiently expressed in Escherichia coli. Molecular identity of rCPI to its natural counterpart has been clearly demonstrated in structural (mass fingerprint), immunological (immunoblot and ELISA inhibition) and biological sense (inhibition of papain proteolytic activity and antifungal activity). Based on the obtained results it is clear that nCPI is a clinically relevant kiwifruit allergen, and as such should be involved in the component-resolved diagnosis of kiwifruit allergy. Standard immunotherapeutic approaches in food allergy are not adequate due to unacceptably high risk-to-benefit ratio, however hypoallergenic forms of molecules with reduced IgE binding potential seem to be promising concept. Diversification of the gene encoding the mature form of the actinidin was done via error prone PCR, followed by cloning of the library into the Aga2 yeast surface display expression system. After three rounds of sorting by FACS, randomly picked colonies were further analyzed by dot blot and ELISA using pool of 6 kiwifruit allergic patients. Compared to both natural and recombinant wild type actinidin two of the mutants exhibited diminished ability to activate basophils..

Allergen immonotherapy in historical perpective definition of an allergen aerobiology allergen nomenclature tree pollen allergens grass pollen allergens weed pollen allergens mold allergens biting insect allergens mite allergens cockroach and other inhalent insect allergens food allergens latex allergens drug allergens, haptens and anaphylatoxins manufacturing and standardizing allergen extracts alergen extract unitage and its clinical significance immunologic responses to immunotherapy efficacy - clinical responses to immunotherapy patient selection for allergen immunotherapy preparing and mixing office extracts administering of inhalent allergenic extracts administering hymenoptera venom and biting insect pharmacoeconomic considerations for allergen immunotherapy potential new forms of immunotherapy recominant allergens, a new form of immunotherapy? unproven and controversial forms of immunotherapy in vitro tests to monitor efficacy of immunotherapy in IgE medicated disease parameters to monitor clinical efficiacy of immunotherapy in IgE medicated disease preventing adverse effects and fatalities associated with allergen skin testing and immunotherapy treatment of anaphylaxis and anaphylactoid reactions suggested forms for use during skin testing and immunotherapy.

The characterization of an allergen is a troublesome and difficult process, as it requires both the precise biochemical characterization of a (glyco)protein molecule and the establishment of its susceptibility to IgE antibodies, as they are the main link to histamine release in some hypersensitivity states (type I allergies). As the characterization of an allergen includes molecular weight determination of the allergenic molecule, its structure determination, physicochemical properties, IgE binding properties of the allergen molecule, and its allergenicity, an overal review of which biochemical and immunochemical methods are used in achieving this goal are presented in this paper. The information on the molecular level on the stuctures of allergens indicates that allergens are considerably heterogeneous protein structures, and that there is no particular aminoacid sequence which is responsible for the allergenicity. Therefore, information gained from detailed structural, functional and immunochemical studies of these intriguing molecules, which nowadays modulate a variety of pathophysiological conditions, would greatly improve our understanding of the underlying disease mechanisms, and the way to handle them. Okarakterisati alergen je težak i mukotrpan zadatak, jer zahteva preciznu biohemijsku karakterizaciju (gliko)proteinskog molekula, kao i ustanovljavanje njegove sposobnosti da vezuje IgE, jer je to glavna spona ka oslobađanju histamina u nekim stanjima preosetljivosti (alergije tipa I). Karakterizacija alergena podrazumeva određivanje molekulske mase, određivanje strukture, fizikohemijskih svojstava, IgE vezujućih osobina i njegovu alergenost. Ovaj rad daje pregled koje se biohemijske i imunohemijske metode najčešće koriste radi postizanja tog cilja. Informacije koje su do sada dobijene o strukturi alergena pokazuju da su ovi molekuli izuzetno heterogene strukture i da ne postoji određena aminokiselinska sekvencija koja bi mogla da predvidi alergenost datog molekula. Međutim, informacije dobijene iz detaljnih strukturnih studija ovih molekula će doprineti našem razumevanju patofizioloških procesa koji su u osnovi alergijskih oboljenja i unaprediće način na koji ih tretiramo.