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Biotransformation and Transport of Xenobiotics in Helminths
Bártíková, Hana ; Szotáková, Barbora (advisor) ; Horák, Petr (referee) ; Machala, Miroslav (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Department of Biochemical Sciences Candidate Mgr. Hana Bártíková Supervisor Doc. Ing. Barbora Szotáková, Ph.D. Title of Doctoral Thesis Biotransformation and transport of xenobiotics in helminths Infectious diseases caused by parasitic helminths are a major problem threatening health of domestic and wild animals and affecting agricultural industry worldwide. Treatment of helminthoses is based on administration of anthelmintic drugs, with benzimidazoles being the most important group. Unfortunately, the irrational use of similar anthelmintics has led to the development of drug resistance in helminths, thus causing a serious problem in the veterinary practice. Possible mechanisms of drug resistance development include changes of pharmacokinetic processes (changes in drug transport or increased drug deactivation), which are based on an increased activity of biotransformation enzymes and transporters in helminths. Understanding the mechanisms of drug resistance and defence strategies of parasites against drugs can prolong the efficacy of current anthelmintics and help to find new strategies for the control of helminthoses. Although drug metabolizing enzymes and transporters of helminths serve as an efficient defence...
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Dual role of CD9 protein in mast cell activation
Machyna, Martin ; Dráber, Petr (advisor) ; Černý, Jan (referee)
Mast cells are well known effector cells in immune system. They have been implicated in such important processes as host defense against bacteria, toxins or parasites. However, in some cases they can develop improper reaction against harmless environmental antigens and thus causing allergies. It is therefore essential to understand signaling events that lead to activation of these cells in order to develop new treatment strategies. Newly prepared rat monoclonal antibody of IgG1 subtype raised against murine mast cells was characterized and found suitable for flow cytometry, immunoblotting and immunoprecipitation. Employing of optimized procedure for immunopurification in combination with mass spectrometry led to identification of its target cluster of differentiation (CD)9 protein. CD9 is a member of large protein family called tetraspanins. Functional studies showed that binding of this antibody to mast cells induced degranulation and early activation events such as increased tyrosine phosphorylation and enhanced levels of free cytoplasmic calcium. Interestingly, subsequent activation of these cells via antigen-mediated aggregation of the high-affinity IgE receptor (FcεRI) led to decreased degranulation, calcium response and tyrosine phosphorylation of several substrates. Importantly, anti-CD9 antibody did...
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Fungi associated with tree roots
Charvátová, Markéta ; Baldrian, Petr (advisor) ; Gryndler, Milan (referee)
In contrast to the bulk soil environment, plant roots represent a habitat with higher nutrient availability due to the supply of photosynthesis-derived C-containing compounds. The roots thus support the life of various microorganisms that use such compounds, but the root-associated microbes in the same time may face a limitation in N and P availability. Unlike bacteria, many filamentous fungi are able to transport these compounds from soil and their mycelia thus typically extend into this environment. Ectomycorrhizal fungi are a typical example of this nutrition strategy. Tree roots produce exudates, that differ in thein composition for each species and they can also differ among individuals. It causes a high diversity of root-associated fungi. Soil is also inhabited by parasitic and saprotrophic fungi. Composition and activity of microbial communisies in the rhizosphere differ from the bulk soil. This work points at the differences of the rhizosphere and builk soil, the importance mycorrhizal fungi and it presents an overview of fungal species that have been found on the roots of spruce - Picea abies. Key words: fungi, mycorrhiza, soil, rhizosphere, Picea abies
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Extra-pair fertilizations and the opportunity for sexual selection in Barn swallows
Michálková, Romana ; Albrecht, Tomáš (advisor) ; Šálek, Miroslav (referee)
Generally it is assumed, that sexual selection drives the evolution of elaborate traits. The strength of sexual selection has traditionally been attributed to varinace in reproductive success among individuals. This variance is mainly caused by the number of mating partners and the number of sired offspring. Both males and females can increase their reproductive success by alternative reproductive strategies - extrapair copulations and intraspecific brood parasitism. In our study we tested the relationship between male ornamentation and success in extra-pair and within-pair paternity and the total male reproductive success in barn swallow (Hirundo rustica), a socially monogamous species with high levels of extrapair paternity. The results suggest more colourful throat to indicate male reproductive success. Dark rusty males were less often cuckolded and they have higher total fertilization success than drab ones. Tail length did not affect neither within- nor extra-pair fertilization success, however, previous studies have concluded that reproductive success of male barn swallows is positively related to the tail ornaments. Our results confirm that extra-pair paternity considerably increase variance in reproductive success among barn swallow males. We show that within-pair success may increase the...
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Working with ethnicity in low-threshold facilities for children and youth
KAŇKOVÁ, Tereza
Everyone brings some dispositions ( factors) into his life, which cannot be swayed. In these factors belong for example skin colour, genetic dispositions or ethnicity. Some of these differences is possiibe to see on the first look and some follow up during the person´s life. It has connection with how we are felt with other people .It is very important how we feel ourselves. On base of some signs we categorize people according to stereotyped used phyziognomic signs. But the identity of the person can be in his conception totally different than it is felt by the other people. Ethnicity is one of the signs which we take in our lifes from the beginning of our existance. Concept of ethnicity correspond with name of race but it´s other meanings follow in the field of culture. Ethnicity is concentrated around same cultur conviction and practice. Forming of the "ethnic groups" consists in sharing of culture signs, which were developed in specific historical, social and political contexts and support the feeling of belonging. The meaning of ethnicity is partly from mythologic origin. Ethnicity is beeing formed on the base of how we express culture identity with the help of the language sings. Therefore is the ethnicity relative and it correlatives with categories of self interpretation and acquire of social status. The dissertation Perception of ethnicity in Low-treshold treatment programe for kids and young people is concentrated on the social workers´s view on social work with ethnicity of users the Low-treshold treatment program. These programes say thet the main group of their users are gypsies. In dissertation I work with the question if the social workers who work with the gypsies are swayed by their ehtnicity. If the social workers have to use some different methods when they work with the gypsies. In the Low-treshold treatment programe work the social workers mostly with children and young people who are social handicapped. Often there are children and young people who only drift aroudn the street. Often there are also children and young people who live in weak social environment and they live in bad areas where they have no opportunity for their personal progress. Quality these children´s life is very low even if we talk about the material aspect or their family background. I concentrate on explaining the word Ethnicity and ethnic minority in the theoretic part. How are the gypsies swayed with their own enthnicity and how do they live in the Czech Republic. Also I talk about the Low-treshold treatment programe for children and young people. Next I write about theories and methods of the social work. I decided for strategy of qualitative research in the empirical part. Here I used the method of questioning, technique of halfstandardized interview. I chose this technique becasuse it is possible to change it and modify during the analysis. Quality dates I recived , I adapted with the help of coding. The social workers in Low-treshold treatment programe made the basic complex for collecting the dates. They have been chosen because of their direct work with the users of the programe. The target of my dissertation was to identify how the user´s ehtnicity sways the work of the social workers. It follows from the resaults that the ethnicity is not important for the social workers. How they said, they feel the user´s ethnicity but in the mutual interaction with users they find some other , more important characteristics. One of the important attribute is the area which the users come from. Next factor is the bahaving of the children. Or the way how are the gypsies behaved. How the social workers said, they feel a little uselessness to change the habits. It followed from the interview that the success of their work is very low. If we talk about some social work specification it is necessary to work with the gypsies according to their social differences. Social workers choose the attitud.
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Black Economic Empowerment - successful strategy or not?
Repatá, Kateřina ; Němečková, Tereza (advisor) ; Adamcová, Lenka (referee)
Republic of South Africa underwent through very significant changes. The discriminatory laws and provisions were cancelled, there were free elections maintained. The government is trying to empower black people. Black Economic Empowerment (BEE) is governmental strategy which seeks to diminish differences between white and so called historically disadvantaged inhabitants. This master thesis provides broad survey of this strategy and analyses its current results.
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The course of microsporidiosis caused by Encephalitozoon cuniculi in experimental model
VOTOČKOVÁ, Tereza
Microsporidia are obligate intracellular parasites causing disease called microsporidiosis. The infectious strategy is a spore - a highly organized cell which gives a rise to the infection of the host. This bachelor thesis records the progress of microsporidiosis provoked by the infection of the experimental model of the species of microsporidia Encephalitozoon cuniculi, namely of the genotype ECIII. The theoretical part includes the familiarization with microsporidia. In this part, there are first records leading to paying more attention to them and initiating their closer examination. This section describes both the biological characteristics and the systematic categorization of microsporidia and the description of a spore as the infectious stage of life cycle, which enables get into the host body where microsporidia reproduce and cause diseases. The sources of microsporidia are mentioned as well as the ways of transmission and microsporidia species most commonly detected in human whose attention is devoted mainly to Encephalitozoon cuniculi - its history and development, the infectious progress in hosts, the detection, and the treatment. The practical part describes the methodical process of the way of monitoring of microsporidiosis on experimental models in a form of laboratory mice of inbred immunocompetent BALB/c and the mice with severe combined immunodeficiency SCID. The mice were orally infected with spores of Encephalitozoon cuniculi genotype ECIII, which was isolated from steppe lemming (Lagurus lagurus). The progress of infection was monitored both on the basis of coprological examination of mouse feces and on the basis of the examination of tissue and body fluid samples. The processing of individual samples was carried out with the aid of molecular diagnostics in the parasitological laboratory AV ČR in České Budějovice. The DNA from individual tissues and body fluids was isolated by using a commercially supplied kit, QIAamp DNeasy Blood & Tissue Kit ( QIAGEN), the feces were treated with a commercially supplied QIAamp DNA Stool Mini Kit (QIAGEN). The principle was to destroy the individual spores and to obtain the pure DNA which was suitable for further processing. Subsequently, the two-step polymerase chain reaction was carried out. providing rapid multiplication of DNA. The next step was a gel electrophoresis which arouses the division of the DNA into individual fragments on the basis of different molecular weights due to the effect of unidirectional electrical current. The fragments were visualized by UV transilluminator at a wavelength of 312 nm linked to a computer. The part of the research was the albendazole treatment of BALB/c mice. The drug was orally administered in the range from 28th to 42nd day after the infection. The results on progress of microsporidiosis through various organs and tissues, including the involved treatment, were recorded in well arranged tables. At the end of this bachelor thesis, there is a summary of the research and its comparison with previous studies. The results showed that microsporidia may be a threat not only for people suffering from immune deficiency, but thanks to their successful survival in the organs of immunocompetent hosts and their ability of activation from the undetectable level, they emphasize the danger of latent microsporidiosis as a risk and life-threatening factor for groups of people undergoing chemotherapy or organ transplantation where the patients can receive inflected graft from a donor.
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Evolution of the tetrapyrrole synthesis in eukaryotes
KOŘENÝ, Luděk
This thesis focuses on the nature of heme metabolism in various eukaryotes. One of the aims was the elucidation of the origin of the unique heme biosynthesis pathway in apicomplexan parasites through a comparative study of their photosynthetic relative Chromera velia combining molecular biology, biochemistry and bioinformatics approach. Using similar strategy, I have also investigated the origin and spatial organization of tetrapyrrole biosynthesis in Euglena gracilis. Based on the phylogenetic data I described the complex evolution of heme metabolism in kinetoplastid flagellates including pathogenic trypanosomes. I revealed that one of them (Phytomonas) does not require heme for viability by the combination of various biochemical and molecular biology experiments and bioinformatic analyses.
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The role of nest in reproduction of the Great Reed Warbler
Jelínek, Václav ; Procházka, Petr (advisor) ; Krist, Miloš (referee) ; Trnka, Alfréd (referee)
7 Abstract Bird nests protect eggs and nestlings, allowing the parents to leave their offspring and subsequently return to them. Their thermoregulatory properties reduce energetic costs of incubation and brooding of nestlings. For all these reasons, nests are key structures for the reproduction of a majority of avian species and as such they should be subject to natural selection. Several hypotheses describing selection pressures which affect the size of nests or some of their parts have been suggested. In my PhD thesis, I investigated some of them in the great reed warbler (Acrocephalus arundinaceus) - an open nesting passerine species with very variable nest size. For this purpose, I used a large dataset of several hundred measured great reed warbler nests, nest enlargement experiments and an experiment with artificial nests. In accordance with previous studies, we did not find that nest size affects the probability of common cuckoo (Cuculus canorus) brood parasitism, while it was influenced by other factors, such as timing of breeding, reed density around the nest and nest visibility from the nearest potential cuckoo perch site. More interestingly, we found that cuckoos adjust their nest-searching strategy in relation to availability of host nests. When host nests were scarce, cuckoos parasitized all of...
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Alternative reproductive strategies in Common Pochards (Aythya ferina)
Šťovíček, Ondřej ; Albrecht, Tomáš (advisor) ; Bryja, Josef (referee)
Conspecific brood parasitism (CBP) is one of few alternative reproductive strategies exmployed by females. CBP is common in birds because of their mode of reproduction (ovipary), which allows relatively simple dumping own eggs to foreign nest. This strategy is especially common in birds with precocial young, such as waterfowl. In this study, common pochard (Aythya ferina) was the model organism for study of CBP via molecular methods. It is the very first evidence of this kind of parasitism in non-colonial and non-cavity nesting duck species, based on detection of parasitic eggs via microsatellites markers. A set of 17 microsatellites markers (10 of these were newly found and specific for our species) was optimised for detection of rate of parasitism and relatedness between females in whole population. It was found that CBP is surprisingly high in our species, the parasitic eggs (39 % of 252 eggs) were found in 89% of nests. Despite the fact that females preferentially nest near relatives individuals, parasitic females parasitize equally nests nearby and nests in longer distance. However, no preferential parasitism of related individuals was found. Cost of parasitism seems relatively low for both, host and parasite, in our species. This is in accordance with previous finding that common pochard's...
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