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Molecular characterization of NADPH oxidase in the gut of the tick \kur{Ixodes ricinus}
KUČERA, Matěj
This thesis focuses on characterization of one member of NADPH oxidases - dual oxidase (DUOX) which has been described as the main factor of epithelial immunity in the gut of model organism Drosophila melanogaster. We have identified an orthologous gene coding for DUOX in the tick I. ricinus and described its tissue expression profile. The DUOX is mainly expressed in the gut of unfed ticks and seems to be downregulated upon artificial microbial infection. A fragment of tick DUOX was prepared as recombinant protein and used for preparation of specific antibodies to be used for further characterization of the enzyme. Our main aim is to highlight the importance of tick DUOX producing the reactive oxygen species and their role in the defence against pathogenic organisms within the tick gut.
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Presence of tick-borne encephalitis in Pribram district with its level of awareness and general attitude to vaccination against it in selected populations
KUCHAŘOVÁ, Eliška
Central European tick-borne encephalitis is a disease belonging to the group of zoonosis, which is transmitted by the tick Ixodes ricinus. It is an acute neuroinfection caused by the RNA virus that can seriously endanger people?s lives. Tick-borne encephalitis is one of the diseases typical for its natural pocket. The most common way of transmission onto a human being is by the bite of an infected tick. However, there have been cases where the contagion took place by consuming unpasteurized milk or dairy products. Although it is a disease which can be easily prevented, vaccination is still very low in our country. If we do not press people to improve this situation, we can expect frequency of tick-borne encephalities will be increased.
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Preparation of recombinant inhibitor of serine proteases from the tick \kur{Ixodes ricinus}
VLNOVÁ, Ivana
Tick serine protease inhibitors could be important anti-tick vaccines targets because of their properties and functions. The aim of this work was to prepare recombinant inhibitor of serine proteases from the tick Ixodes ricinus in baculovirus expression system. Two tick saliva proteins of the serpine superfamily were selected for this purpose and transformed into plasmids. One recombinant protein was expressed in baculovirus expression system, purified and its biochemical analyses were done.
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Vaccine potentiality of cystatin from tick \kur{Ixodes ricinus}
HARCUBOVÁ, Adéla
Ticks belong to the ectoparasites which are dangerous for the human beings because of the transmission of bacterial, viral, and protozoan pathogens. The development of a vaccine against tick is very important. Cystatins play important role in tick digestion and they have an immunomodulation effects. For this reason cystatins are possible candidates for this vaccines. This thesis focuses on experimental mice vaccination with recombinant cystatin from Ixodes ricinus.
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The effect of tick´s serpin IRS-2 on dendritic cells activated by TLR4 ligand
POSPÍŠILOVÁ, Šárka
IRS-2 is the inhibitor of serine proteases from the Ixodes ricinus tick. My task in this thesis was to find out the effect of the IRS-2 on dendritic cells activated by TLR4 ligand or by Borrelia afzelii. This effect was studied on several levels. I focused on the cytokine production, the expression of costimulatory molecules and cell signaling pathways. The results show that the IRS-2 may inhibit the expression of costimulatory molecules CD-80 a CD-86 on the cell surface, but this finding needs to be confirmed again. The production of cytokines was not affected by the IRS-2. The effect of the IRS-2 on the activity of p38, Erk1/2 nor NF-?B in LPS stimulated cells vas not observed. The fosforylation of STAT 3 in cells activated by the B. afzelii was lowered by the IRS-2.
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Dynamics of digestive enzymes in the gut of ticks \kur{Ixodes ricinus} during blood feeding on the host.
KONVIČKOVÁ, Jitka
Proteolysis of ingested blood proteins is a crucial physiological process in ticks. This thesis focuses on profiling of dynamics of digestive enzymes in the ticks Ixodes ricinus during the course of blood-feeding on the host and during digestive phase after detachment from the host. We have measured the activity of digestive peptidases (using specific fluorescent substrates and inhibitors). The enzyme activities were negligible at the early stage of bloodfeeding than increased dramatically towards the end of the feeding period and dropped down immediately after detachment from the host. The activity of most enzymes increased again during the digestive phase before oviposition of females or metamorphosis of nymphs. The protein level of each peptidase was confirmed by immunolocalization using Western blot analyses.
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Characterization of a defensin of the tick \kur{Dermacentor marginatus}
LEŠTINOVÁ, Kateřina
Antimicrobial peptides (AMPs), as a part of innate immune system of ticks and other living organisms, are able to eliminate pathogens. In ticks the most important group of AMPs is defensin family. In this work, defensin from the tick D. marginatus was studied. The defensin gene was isolated from D. marginatus fed females. Using RT-PCR the gene expression was detected in salivary glands and mitgut. Recombinant protein was expressed in the procaryotic expression system, purified and tested for its antimicrobial activity. Specific polyclonal rabbit antibodies (anti DR IgG) were prepared and tested for their specifity and sensitivity.
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The transmission of significant pathogens activated by tick saliva
HUSPEKOVÁ, Helena
Ticks are one of the most common ectoparasites in the world and they are vectors of many tick-borne diseases. These diseases are caused by pathogens, including bacteria and viruses. The main point of the thesis is focused on enhancement of pathogens transmission, (especially Borrelia burgdorferi sensu lato, the causative agent of Lyme disease) by tick saliva or salivary glands extracts, and I tried to refer the main molecules, which have immunomodulatory or inhibition effect on the immunity of host. This effect can lead to the enhancement of pathogens transmission and contributes to their extension in host. The second part of the thesis is based on molecules, which can lead to the creation of vaccines against tick-borne diseases.
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