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Exploration of the tick-Borrelia molecular interactions by employing the transcriptomic approaches
MAHMOOD, Sazzad
Along with climate change and increased sharing of habitat, ticks are coming into more frequent contact with humans. The hard tick Ixodes scapularis and Ixodes ricinus are known disease vectors in Northern America and Europe, respectively. Along with many other pathogenic microorganisms, these ticks spread Borrelia sp. by ectoparasitic blood feeding. Borrelia afzelii is the major European Lyme disease pathogen spread by I. ricinus. Our study focuses on differential gene expression in I. ricinus salivary gland and midgut, induced in the nymphal stage by B. afzelii infection. Tick genes upregulated by infection are considered to play essential roles for the acquisition, persistence, and transmission of Borrelia. We have determined 32,897 full length sequences of tick mRNA from B. afzelii infected/noninfected tick salivary glands and the whole body. In addition, we have obtained MACEseq (Massive Analysis of cDNA Ends) from both midgut and salivary glands while the nymphs were non-infected or infected with B. afzelii during three different phases of blood-feeding. From the MACE database, we obtained 250-500 bp 3'-end sequences with raw quantitative expression values. Total reads, unique sequences and protein coding tick genes from midgut samples were 38,199,641, 88,825 and 24,276, and from salivary gland were 74,651,134, 93,096 and 26,179, respectively. After filtering, using several criteria, expression was validated by qPCR. Hence, the validated genes may most likely interact with Borrelia in its acquisition, persistence, or transmission to the vertebrate host. In our study, RNA interference approaches and vaccination were implemented in order to investigate the impact of upregulated tick midgut and salivary gland genes on Borrelia transmission to C3H mice.
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Borrelia - host interactions: zoom in on the big picture.
STRNAD, Martin
The thesis was written with the intention to bring together cutting-edge imaging methods and applications in order to illustrate how imaging can answer pathogenesis-related questions in Lyme disease at various resolution scale. Correlative light and electron microscopy, atomic force microscopy-based single-molecule force spectroscopy and solution nuclear magnetic resonance have been used to shed light on the underlying mechanisms associated with Lyme disease Borrelia infection. Specifically, the key molecular players and interactions responsible for the variance in the pathogenicity and disease outcome of Borrelia species have been studied. The rationale behind such studies was highlighted by review articles, which are part of the thesis.
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Knowledge of Pupils Secondary School in the Area of Selected Zoonoses
Svobodová, Barbora ; Pavlasová, Lenka (advisor) ; Říhová, Dagmar (referee)
The subject of the dissertation deals with the secondary school pupil's awareness of zoonoses. Zoonoses are the diseases transmited from animals to human beings. Those are one of the topics of biology study currently. The main aim of the dissertation is theoretical definition of the matter in the first place. For knowledge testing have been chosen these four illness: Toxoplasmosis, Lyme boreliossis, Rabies and Creuzfeldt-Jakobo disease. With these chosen illness the main research goal is to provide a comprehensive collection of information about the details such as disease transfer, spread of the disease, disease development, cure and avoidance. The research part of the dissertation tries to find out the extent of knowledge about the chosen kinds of zoonoses among the pupils at specialized secondary schools. As a tool of quantitative research has been used a pupil's didactic test. As based on the evaluated results we may say that the pupil's knowledge at explored schools is relatively satisfactory.
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Students' knowledge of upper primary school about diseases transmitted by ticks
Michálková, Pavla ; Hanušová, Jaroslava (advisor) ; Thorovská, Alena (referee)
The main research problem concerns the characteristics of tick-borne disease and determine the quantity and quality of information that students get in health education and biology of these diseases. The theoretical part deals with the history, etiology, clinical features, diagnosis, treatment and prevention of tick-borne encephalitis and Lyme disease. It also discusses the general problems of tick and other diseases that ticks carry and cause. Also is mentioned the inclusion of tick-borne diseases in the educational program. The practical part is devoted to the awareness of upper primary school students and changes in their knowledge about the issue during their elementary education. The aim of this thesis is to determine how the students are informed about diseases transmitted by ticks, especially about Lyme disease and tick-borne encephalitis. This method is used questionnaire, which includes questions regarding this issue. Although the results of the research show a certain degree of knowledge of students at upper primary school, especially in some classes is unsatisfactory. Based on research, there is provided a recommendation for pedagogical practices. KEYWORDS: tick, tick-borne encephalitis, Lyme disease, knowledge, disease, upper primary school
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Molecular Docking Study Of Potential Drug Candidates Against Borreliosis
Koščová, Pavlína
Lyme borreliosis is one of the most common tick-borne infections, for which there is an extensive need to find a new drug. For this purpose our in silico docking study was carried out to identify drug-likeness of chosen small molecules – potential borreliosis drugs. Its results revealed that BesA compound (C2 form) – a membrane fusion protein present in Borrelia burgdorferi, can play a significant role as a possible drug target compound and therefore it should be further examined in development of potential drugs for Lyme borreliosis treatment.
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Metamorphoses of \kur{Borrelia burgdorferi} sensu lato spirochetes: from dormant to motile forms
MORÁVKOVÁ, Veronika
The aim of the study was to obtain and elaborate information focused on metamorphoses of Borrelia burgdorferi sensu lato (s.l.) spirochetes. The research included detection of various stress conditions for production of dormant forms of Borrelia burgdorferi s.l. spirochetes in vitro. Subsequently, metamorphoses from dormant to motile stages was observed under favourable conditions. Proper PCR method for primers aimed to detect dormant forms of Borrelia was delevoped. The infectious potential of dormant spirochetes of Borrelia burgdorferi sensu stricto (s.s.) complex was observed in vivo. Transformations of spirochetes have also been observed in real time and individual stages have been recorded.
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Comparative Analysis of Common and Unique Targets in Drug Resistant Strain of Borrelia Burgdorferi
Koščová, Pavlína
The number of drug-resistant strains of Borrelia burgdorferi necessitated the identification of potential drug targets specific to the strain of interest. The chromosomal and plasmid genes of B. burgdorferi strain B31 were compared with erythromycin-resistant B. burgdorferi strain N40 to find common (core) and unique (strain-specific) genes in present study. In silico analysis of genomic data showed total number of unique genes higher in strain N40. The presence of higher number of unique genes in N40 signifies their role in drug resistance mechanism. Furthermore, human proteome was compared with proteome of these strains to find target protein specific to the strain of interest and not present in host. In conclusion, identification of unique genes in these strains provided on differences in drug resistance potential.
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Studie antimikrobiálního účinku včelího jedu na druhy borelie z komplexu \kur{B. burgdorferi} sensu lato: izolace genu kódujícího antimikrobiální peptid melitin z jedové žlázy včely medonosné \kur{(Apis mellifera)}
DAVIDOVÁ, Marie
The incidence of tick borne diseases has drastically increased over the past few years. Control of tick borne pathogens, especially spirochetes from \kur{B. bugdorferi} sensu lato complex, is complicated due to a lack of vaccines against them. The emergence of bacterial strains to well established and widely used antibiotics has become a serious public health concern. Analysis of new classes of antimicrobial peptides became the subject of new investigation due to their efficacy against disease causing pathogens. Among possible candidates, melittin from honey bee venom is of particular interest. The aim of this work was to study the effect of whole bee venom on Lyme borreliosis spirochetes. The minimum concentration of the venom that possesed the inhibitory effect on borrelia growth was identified. The gene enconding melittin was isolated from bee venom gland and recombinant melittin was produced in bacterial expression system. The effect of recombinant melittin on live borrelia culture was studied.
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Gene expression and infectivity of \kur{Borrelia afzelii} in the course of tick feeding
POSPÍŠILOVÁ, Tereza
Borrelia afzelii differential gene expression in the course of tick blood-feeding, and during chronic infection in mice was studied. Temperature effect on B. afzelii gene expression and infectivity was investigated. Infection rates of mice immunized with B. afzelii tick gut antigen at various stages of tick blood-intake were analyzed. This work was funded by the Grant Agency of the Czech Republic, Project No. 17-27393S to Radek Šíma.
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