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Section 6. Medical science
Abdullaeva Dilafruz, Tashkent Institute of Postgraduate Medical Education, assistant E-mail: [email protected]
The new features of in vitro allergodiagnostics of food allergy in hot climate conditions
Abstract: We implemented immunoblotting panels of allergens into clinical practice for diagnosis of food allergy and cross-reactions to food, pollen, fungi and latex. These panels are widely used in the republican allergy center, allergy departments of Khorezm, Bukhara, Surkhandarya, Kashkadarya, and also in clinical centers of Samarkand and Andijan regions. Keywords: Food allergy, allergenic proteins, allergenic foodstuff, diet therapy.
Introduction
There is presently a tendency to an increasing frequency of allergic diseases. About 30 % of the world's population suffers from various forms of allergy [2; 3].
Currently, there is no accurate food allergy prevalence data for our country. On the one hand, this is due to numerous disease manifestations that affect many organs and systems. On the other hand, this is connected with difficulties with accurate diagnosis verification in clinic practice. There are approximately 220-250 million people suffering from allergy diseases [11].
The objective of our study was an examination of the food allergy diet therapy in a view of food properties sensitizing by "in vitro" methods.
Material and methods
We have introduced "Rida qLine" allergy panels (R-Biopharm, Germany) to the republican allergy center, allergy departments of Khorezm, Bukhara, Surkhandarya, Kashkadarya and in clinical centers of Samarkand and Andijan regions for food allergy and cross-reactions to food, pollen, fungi and latex diagnosis. This test is based on the principle of immunoassay on a nitrocellulose membrane (immunoblot) for a quantitative determination of allergen's specific IgE antibodies of the patent's serum and plasma. The allergens that correspond to the panel composition are applied to the surface of nitrocellulose membranes. Results are expressed in IU/ml and RAST (radioallergosorbent test) classes (0-6). There are four different panels. All results with IgE greater than 0.35 IU/ml were considered to be positive. Food panels contain the most common in our region food allergens and their mixes.
Results and discussion
Allergy diagnosis is difficult due to the lack of uniform methodological approaches and diagnosis methods that allow to identify allergy development mechanisms. In pseudo allergic reactions, it is impossible to detect specific antibodies that participate in true allergic reactions. The reaction is dose dependent, and unlike to allergic reactions, pseudo allergic reactions invoke during the first contact [4].
Taking into account climatic- geographical features and food traditions of the population, we suggested another allergens panel composition for our country. This composition consists of 2 types of food allergens, children and mixed allergen panels. Every panel has 20 allergens and 5 standards. All standards are calibrated by the reference to international protocols «1st WHO IRP 67/86 for
human IgE». The results are expressed in IU/ml (0-100 IU/ml) and in RAST classes (0-6).
In immunoblot panels we included typical for our region food products (strawberry, lemon, peach, cow's milk, egg, tomato, pea, peanut, carrot, walnut, honey, chicken meat, beef, mutton, horse-meat, cherry, wheat flour, oak flour, buckwheat flour, banana, yogurt, gluten, mixtures of nuts, fruit, vegetables, meat, legumes), epidemical (bovine serum albumin, animals' epithelium, plumage), fungi (Cladosporiumherbarum, Penicilliumnotatum, Candida albicans, Alternariaalternata, смеси Aspergilli, Mucormucedo, Rhizopusni-gricans), professional (latex) and pollen (tree pollen, meadow pollen, weeds pollen) allergens.
We tested the blood of 65 children at the age under 18 by the use of a pediatric panel and 32 adults at the age from 18 to 60 by the use of a food panel. According to our research results, the most allergic food products in a hot climate conditions among children are buckwheat flour in 26.1 % cases; gluten — 24.6 %; nuts, wheat flour — 16.9 %; rice, yogurt — 13.8 %; cow's milk — 12.3 %; eggs — 10.7 %; oat flour — 4.6 %. 23.1 % of patients under 18 had reaginic antibodies E to a grass mix (meadow fescue, timothy, rye) and 15.4 % to birch. We have not detected any antibodies to banana, meat mix (beef, lamb, pork) and soya protein.
In accordance with the results, 18.7 % of adult patients (18-60) had allergen specific antibodies E to pea; 15.6 % to tomato, carrot; 12.5 % to honey, chicken, wheat flour, potato, apricot; 9.3 % to watermelon, strawberry; 6.2 % to goat's milk, walnut, orange, peach, egg yolk, apple, pineapple, celery, beef, lamb, cow's milk, egg white; 3.1 % to sesame. There have not been detected any antibodies above normal to coffee.
Furthermore, we tested 90 patients by the use of a mix panel. By analyzing the results, we got the following percentage. 24.4 % of patients had antibodies to nuts (peanut, hazelnut, almond, Brazil nut, coconut, walnut), 26.6 % to milk mix (casein, cow's milk, milk powder), 18.8 % to birch, 17.7 % to mould fungi (Mucor mucedo, Rhizopus nigricans), aspergillii mix (Aspergillus flavus, Aspergillus fumigatus, Aspergillus niger, Aspergillus versicolor) and 12.2 % to latex. All the latex sensibilized patients (100 % cases) had reaginic antibodies over 1 RAST to birch and nuts; 45.4 % of them had antibodies to mould fungi.
After allergy product determination, we offer an elimination of the cause allergen out of patient's ration. The knowledge of food
The new features of in vitro allergodiagnostics of food allergy in hot climate conditions
protein's structure and cross properties with other allergens groups helps dietitians and patients with a creation of a diet ration. Structural studies often reveal features of biologic importance that might not be apparent from biologic assays. Allergens belong to protein families with diverse biologic functions that can be summarized as follows: Indoor allergens (enzymes (especially proteases), ligand bilding proteins or lipocalins, albumins, tropomyosins, and calcium-binding proteins), pollen allergens (pathogenesis-related proteins, calcium-binding proteins, proteins, pectate lyases, ^-expansins, and inhibitors), plant and animal food allergens (LTP- lipid transfer protein profilins, seed storage proteins, and tropomyosins) [8].
Plant food allergens include the seed storage 2S albumins found in tree-nuts and seeds, the defense-related nonspecific lipid transfer proteins found in soft fruits and vegetables, and cereal a-amylase/trypsin inhibitors [5; 8]. The second major superfamily of plant food allergens, the cupin family contains 2 groups of seed storage proteins called vicilins and legumins, which are important peanut and tree nut allergens. The profiling and Bet v 1 family includes tree pollinosis-associated food allergens with low stability that induce symptoms of the oral allergy syndrome. These 4 protein families contain approximately 65 % of all plant food allergens. Of the remaining 27 allergen-containing protein families, more than 50 % harbor allergenic proteins of the plant defense system or pathogenesis-related proteins, such as the cysteine proteinases, thaumatin-like proteins, or chitninases [5; 9].
The most important animal food allergens are present in milk, egg, and seafood. Mammalian milk allergens are found predominantly in 3 protein families, a-lactalbumin, which is essential for milk production, is a member of glyosyl hydrolase family. ^-lactoglobulin is a lypocalin, and the casein family harbors the major constituents of milk. Ovomucoid, the most important egg allergen, is a serine protease [9].
Phytalbumin and animal protein dominated amongst the food allergy ill in causal food allergens spectrum. Cow's milk and chicken protein are the most common cause of food allergy among animal proteins, and cereals proteins are widespread among phytalbumins, especially for children. Cereals intolerance can appear both as a food allergy and as a gluten enteropathy that often makes diagnosis and treatment complex. It was detected that 30 % of allergic children had allergen IgE-antibodies characterized by digestive system and skin symptoms [1].
Fruit and vegetable allergies are the most prevalent food allergies in adolescents and adults. The identification of the allergens involved and the elucidation oftheir intrinsic properties and cross-reactivity patterns has helped in the understanding of the mechanisms of sensitisation and how the allergen profiles determine the different phenotypes. The most frequent yet contrasting fruit and vegetable allergies are pollen-food syndrome (PFS) and LTP syndrome. LTP syndrome results from a primary sensitisation to LTPs, which are stable plant food allergens, inducing frequent systemic reactions and even anaphylaxis [6]. PFS usually presents with pruritus and swelling of the mouth and throat during or just after ingestion of fresh, uncooked fruits and vegetables [7]. LTP can be an important cause of allergy given their stability and high degree of protein sequence homology. LTP allergy should be considered when anaphy-laxis is inconsistent, such as in patients who can tolerate fruit pulp but react to fresh whole fruit juices [10].
Fruit family Rosacea is one of the most common fruit allergens in hot climate conditions (peach, apple, strawberry, apricot). Allergy on apples was combined with allergy on birch pollen for 25 % of patients. Presumably, this combination was observed due to some
similarities between allergens, and the allergy was expressed in a form of allergic rhinitis, bronchial asthma. Only 2.5 % of patients had oropharyngeal affection [1].
In our case 12.5 % of patients had sensibilization to walnut and 5 % to peanut. It should be noted that allergic properties ofpea-nut increase by culinary processing. Two main peanut allergens are coctostable and resistant to digestive enzyme's actions. It ought to be observed that allergy to peanut and to other legumes often begin in the childhood and remain constant for a long time. The allergy can be observed even in adulthood [1; 4].
The lack of uniform diagnostic criteria, the variety of clinical manifestations of food intolerance, conditioned by different (immunological and non-immunological) mechanisms and of development of allergic cross-reactions for food products, often leads to misdiagnosis, and therefore to wrong diet therapy.
Eliminating diet prescription tactics depends on examination period, severity of clinic manifestations, causal allergens spectrum, and patient's age. Diet therapy effect is determined not only by an accurate identification of food product that causes the development of allergic reactions, and by its ability to cross reactions, but also by a correct determination of reasonability and elimination duration depending on allergic properties of food allergens.
The usage of "in vitro" methods in specific diagnosis may be the only possible measure in cases when, for instance, skin tests are dangerous or impossible (with an occurrence of skin allergic processes, for example, atopic dermatitis, urticaria) in the period of bronchial asthma exacerbation or pollinosis with no option of antihistamine withdrawal after having an anaphylactic shock in the past. In addition, "in vitro" methods are useful when clinical methods appeared to be not informative enough. The test is informative in the period of allergic diseases exacerbation, and the test does not need a special patient's preparation. Moreover, there is no risk of systemic reactions occurrence and allergic diseases exacerbation because of the test. The reason is that a contact between a patient and an allergen is eliminated.
There are several advantages of "in vitro" method over skin tests: the examination can be conducted in the period of exacerbation; there are no age restrictions (it is possible to use "in vitro" method for infants, pregnant women, in the period of lactation and for elderly people); there is no necessity in antiallergic treatment withdrawal the period of examination; it is possible to detect a sensibilization to a number of allergens at once.
It is considered to be a disadvantage of the method that the "in vitro" examination takes more time than skin tests. Furthermore, the method may show a negative take for the patients with atopic diseases due to a short life span of circulating IgE or because of IgG-condi-tioned reaction occurrence. Before a specific allergic diagnosis method prescription, an immunological mechanism treats of allergic reaction realization needs to be taken into consideration in order to determine a list of possible allergens. For this purpose, a doctor has to thoroughly scrutinize an anamnesis, clinic manifestations of a disease in the past and the presence. Analyzing this the doctor should assume allergens spectrum and type (types) of patient's immune system.
Conclusion
All things considered, it is reasonable to note the actuality and availability of the usage of immunoblot allergens panels for eliminating diets planning. Diets therapy is based on the detailed analysis of a composition and allergic properties of the food product. Apart from that, climatic diet features are taken into account. "In vitro" diagnostics is safe for patients, and it creates conditions for an early causal allergens diagnosis and for a latent sensibilization determination.
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Akhmedova Sayora Muhamadovna, Samarkand State Medical Institute, Uzbekistan, Senior researcher of Department Human Anatomy E-mail: [email protected]
Histological structure of rat heart in the early stages of ontogeny
Abstract: In order to identify the features of morphogenesis in postnatal ontogenesis examined histological structure of different departments and shells in the heart of rats 1-22 days after birth. There alternating periods of acceleration and deceleration of the growth rate increase in the thickness of the atrium and the ventricles. The thickness of the endocardial and epicardial increases less significantly. The growth rate of the ventricular myocardium thickness observed in rats 6 and 16 days of age. Structural changes occur due to the growth of the organism. A special feature of the structure and topography of the heart microvessels is their distribution in the course of the cardiomyocytes and the relationship with the fibrous structures of the connective tissue of cardiomyocytes.
Keywords: heart rats, postnatal ontogenesis, cardiomyocytes, fibrous structure of the atrium and the ventricles.
Relevance of the topic
Analysis of recent data shows that cardio vascular disease remains the leading cause of death in the developed world [1; 2; 3; 5]. Heart frequently undergoes pathological effects of endogenous and exogenous stimuli, resulting in not only a violation of its function, but also develop various pathological processes in the body. Heart animals and humans and can be adapted to change depending on lifestyles and total body burden [7]. The age structure of the heart is of some importance in the disclosure of certain pathological processes developing in it. Also in the pathogenesis of many diseases of the cardiovascular system it plays a major role component connective infarction, it is the most dynamic component in the heart during morphogenesis and pathogenesis [4; 6; 8]. Therefore, a deep and comprehensive study of the structure of the heart walls and connective tissue component, is relevant and reasonably identified the choice of the direction of our research. The heart of the rat is a convenient model for experimental studies, as well as to obtain enough material to study.
Materials and methods
Studied 50 heart rats at 1, 6, 11, 16, 22 hours after birth. In each age group studied 10 rats heart. The animals were kept under standard vivarium conditions at t 21-22 °C and natural photoperiod on a normal diet. Experimental studies were carried out in accordance with the "Rules of work with the use of experimental animals". Slaughter of rats was performed under ether anesthesia. After
opening the chest and abdominal cavities of the material was fixed in 12 % neutral formalin solution, then isolated heart. After fixation, the heart was withdrawn, and washed for 1 -x days in running water. After the body was carried out by increasing the concentration of alcohols. Then the organ divided by the atria and ventricles, and embedded in paraffin. Histological sections 8-10 microns were prepared from paraffin blocks. Apply stain sections with hematoxylin and eosin, van Gieson techniques, Weigert and impregnation method Foote modification Yurina. Morphometric measurements of the wall thickness of the atrium and ventricle are made via the line by increasing ocular ob. 90 microscope ok. 7. Mathematical processing of data was performed using Microsoft Excel 2010 application programs in the topic of descriptive statistics, determining the standard deviation, the arithmetic mean M, the average error of the relative values of m. Authenticity of the received data at P < 0.05.
The wall of the atrial heart rats is characterized by uneven thickness. Analysis of the table shows that the thickness of the left and right atrium for the entire study period gradually increases. The 6-day age rats in the left and right atrial wall thickness increase compared to infants is 14 % and 11 %. At 11 days of age, compared to 6 daytime pups left atrial thickness increase of 21 % and 14 % of the right atrium. In 16-day old rats 18 % and 13 %. The growth rate of the left and right atria thickness in rats 22 days of age increased by almost the same and amounts to 20 % and 19 %. Thickness endocardial right
