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Proteomic analysis of lysozyme and lysozyme-like proteins of synanthropic mites
Chum, Tomáš ; Erban, Tomáš (advisor) ; Mikeš, Libor (referee)
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...
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Nutritional biology of synanthropic mites (Acari: Acaridida)
Erban, Tomáš ; Smrž, Jaroslav (advisor) ; Grubhoffer, Libor (referee) ; Šustr, Vladimír (referee)
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...
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Proteomic analysis of lysozyme and lysozyme-like proteins of synanthropic mites
Chum, Tomáš ; Erban, Tomáš (advisor) ; Mikeš, Libor (referee)
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...
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Gluten-free diet and current situation in the restaurants
KUCHAŘOVÁ, Miroslava
The theses called "Gluten-free diet and current situation in the restaurants" is focused on the issues associated with the availability of gluten-free meals in eating facilities of various types and with the possibilities of customers with gluten intolerance when eating out. In the first theoretical part, a "celiac" disease is described. It is compared with other forms of gluten intolerance there, too. Next part is dedicated to food allergen labelling because this topic is very close to the gluten-free diet. This part also characterizes various types of eating facilities and I depicts the territories where my research was made (the city of České Budějovice and the city of Klatovy), finally. The aim of this thesis was to take a close look at the offer of gluten-free meals in various types of eating facilities in the above mentioned cities and to examine the cooks´ knowledge of the principles of gluten-free diet. Here I compared nutritional values of gluten-free diet one-week menu with balanced diet recommendations, too. At the very end of the theoretical part I transformed some meals from restaurants´ standard menus into gluten-free version. In the practical part, a combination of quantitative and qualitative research methods was applied. There were 20 eating facilities included in the research 10 in the city of České Budějovice and 10 in the city of Klatovy. The quantitative method was used to analyze the menus of the given eating facilities, the qualitative method, on the other hand, in the form of semi-structured interviews with the representative of these facilities. The results of the analysis of the menus were put into the Excel chart. The interviews were interpreted through mind maps. When compiling a one-week gluten-free diet I used a computer program "Nutriservis Professional". Finally, I compared the data from the above mentioned program with the figures recommended for the balanced diet. The analysis of the restaurants´ menus brought the following results normally, eating facilities don´t offer as many gluten-free meals as meals containing gluten. However, a number of naturally gluten-free meals in restaurants´ menus is not so bad. Moreover, the analysis of the interviews suggests that there is no problem to agree with the cooks on preparing a gluten-free meal if it is not in the menu. What is not satisfying is allergen labelling in restaurants´ menus and the cooks´ knowledge of gluten-free diet, too. The comparison of gluten-free and balanced diets implies that they do not differ too much as for the basic nutrients, energy, proteins, carbohydrates or fats. To conclude, this fact proves that gluten-free diet is adequate to the balanced diet.
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Allergen-specific immunotherapy from the perspective of nurses.
BEJTICOVÁ, Lenka
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.
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Allergic diseases in relation to lifestyle risk factors by children in younger school age in the Region of Karlovy Vary
RYBOVÁ, Eliška
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.
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