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Recombinant aspartic proteases of blood-feeding parasites
Váchová, Jana ; Konvalinka, Jan (advisor) ; Entlicher, Gustav (referee)
The blood fluke Schistosoma mansoni and the hard tick Ixodes ricinus produce an aspartic protease cathepsin D which initiates degradation of hemoglobin, their key nutrient. First, in the presented work, the protocol for refolding and activation of the zymogen of cathepsin D from I. ricinus (IrCatD) was developed and optimized. In acidic pH the propeptide of IrCatD zymogen was removed by an auto-activation mechanism. Further, a kinetic assay with fluorogenic substrates was employed to study functional properties of IrCatD including pH optimum, substrate and inhibition specificities. Second, two isoforms of cathepsin D from S. mansoni (SmCatD) were produced using recombinant expression in E. coli. These recombinant proteases were isolated from inclusion bodies using affinity chromatography under denaturating conditions, and protocol for their refolding was developed and optimized. The studied aspartic proteases are pharmacological targets: inhibitors of SmCatD represent potential chemotherapeutics for the treatment of schistosomiasis, and IrCatD is a candidate antigen for the development of novel anti-tick vaccines.
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The Castor Bean Tick (Ixodes ricinus) in Education
Němec, Radek ; Říhová, Dagmar (advisor) ; Hanel, Lubomír (referee)
The aim of this work is to point out the interconnectedness of human life with parasitic organisms, especially with the Castor bean tick (Ixodes ricinus). This tick forms an integral part of our rich invertebrate fauna and has become an increasingly frequent interest of media in the last few decades. This thesis attempts to bring closer the relationship between tick and man from different perspectives, and to reveal essence as well as pitfalls of tick life. For many people, this arthropod represents an insurmountable resource of aversion and fear. Hence, an integral part of the thesis is the introduction of prevention methods and treatment of parasitic diseases transmitted by the tick, as well as monitoring of new trends in the therapy and development of vaccines. Also, the text offers a proposal how to bring the tick closer to the pupils in teaching Biology. KEYWORDS The Castor bean tick, parasite-host relationship, life cycle, encephalitis, borreliosis, collection methods
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Methodology of detection of tick-borne pathogens and statistics of their occurrence in the Czech Republic
BROŽKOVÁ, Zdeňka
This bachelor thesis deals with tick-borne transmitted pathogens and with ticks alone. In general, it focuses on ticks species, that are able to infect people. The Thesis is divided into three parts. The goal of the theoretical part is to refer that borreliosis and tick encephalitis are not unique infection diseases endangering human health. In this thesis, infection diseases are divided into bacterial, viral and protozoan. The core of my work is the empirical part, that uses the method of tick-borne pathogen diagnosis. In the practical part I have tried to detect pathogens in ticks by means of PCR and RT-PCR. The third part is based on statistical indexes, where I have compared laboratory results from years 2008-2018. I have analysed the results for a period of 10 years of testing and simultaneously the results of various infection diseases. Statistical data were compared with reports of infection diseases occurrence from the National Institute of Public Health in the time period.
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Detekce a kvantifikace inhibitorů proteáz v klíštěti \kur{Ixodes ricinus} pomocí monoklonálních protilátek
VANÍČKOVÁ, Martina
Inhibitors of proteases in tick saliva play an important role during tick feeding. Tick saliva contains a wide range of bioactive components which are able to modulate host imunity. Therefore, ticks are able to feed for a long time and transfer tick-borne diseases pathogens. The risk of transfer can be significantly reduced by deactivation of theese protease inhibitors. In this study I made monoclonal antibodies for detection and quantification of two serine protease inhibitors in tick saliva and other tick-body parts.
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Initial knowledge of kindergarten pupils about natural phenomena.
FULMEKOVÁ, Kateřina
This thesis focuses on knowledge and notions children attending Kindergarten have about certain Invertebrate species, specifically an ant, an earthworm, a hard-bodied tick and a butterfly. A special interest is being taken in observing artistic expressions of the kids where these animals are concerned. The thesis also describes approaches used to educate children on the Invertebrate in Kindergarten and later in elementary school
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Recombinant aspartic proteases of blood-feeding parasites
Váchová, Jana ; Entlicher, Gustav (referee) ; Konvalinka, Jan (advisor)
The blood fluke Schistosoma mansoni and the hard tick Ixodes ricinus produce an aspartic protease cathepsin D which initiates degradation of hemoglobin, their key nutrient. First, in the presented work, the protocol for refolding and activation of the zymogen of cathepsin D from I. ricinus (IrCatD) was developed and optimized. In acidic pH the propeptide of IrCatD zymogen was removed by an auto-activation mechanism. Further, a kinetic assay with fluorogenic substrates was employed to study functional properties of IrCatD including pH optimum, substrate and inhibition specificities. Second, two isoforms of cathepsin D from S. mansoni (SmCatD) were produced using recombinant expression in E. coli. These recombinant proteases were isolated from inclusion bodies using affinity chromatography under denaturating conditions, and protocol for their refolding was developed and optimized. The studied aspartic proteases are pharmacological targets: inhibitors of SmCatD represent potential chemotherapeutics for the treatment of schistosomiasis, and IrCatD is a candidate antigen for the development of novel anti-tick vaccines.
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