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Motility and virulence of bacterial pathogens in patients with cystic fibrosis
Benešová, Nikol ; Melter, Oto (advisor) ; Smělíková, Eva (referee)
Cystic fibrosis is an autosomal recessive disease that mainly effects the Europoid race. The disease is caused by a mutation in the cystic fibrosis transmembrane conductance regulator gene. This gene encodes the formation of chloride channel proteins. The disease leads to the formation of highly viscous secretions from the exocrine glands. The most commonly affected organ is the lung, which accumulates mucus and bacteria. Bacteria that colonize the lungs of patients with cystic fibrosis are exposed to many stress factors such as large amounts of mucus, oxidative stress, antibiotics and immune system of the host. That leads to the selection of mutants that are better adapted to the environment of the lungs. In acute infection, one of the important virulence factors for bacteria is their motility. Motility is provided by flagella and allows the bacterium to move to sites with higher nutrient content and to colonise different parts of the lung. In chronic infection, we can see changes in the genotype and phenotype of bacteria. Nonmotile mutants are selected because they activate the immune system less than motile bacteria, that helps in the persistence of bacteria in the lungs. Chronic infections lead to a higher mortality rate. The aim of this bachelor's thesis is to compare the motility of bacteria...
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Bacterial adhesion to surfaces
Budil, Jakub ; Beranová, Jana (advisor) ; Kuthan, Martin (referee)
Bacterial adhesion is a process of attachment of a planktonic cell to the surface and the necessary step for further biofilm formation. Most bacterial species are capable of biofilm formation. Bacteria within biofilm exhibit increased resistance to antibiotics and other external factors, which makes the eradication of adhered bacteria rather difficult. Bacterial adhesion is very complex process affected by many physical and chemical factors as well as by the adhesive molecules present on the substrate and cell surface. Accurate description of these factors enables to design appropriate modifications of both the substrate and the environment with the aim to fabricate the antiadhesive and antibacterial surfaces or to increase the efficiency in bacteria-based biotechnological processes. Whereas in medical applications the main goal is to decrease bacterial adhesion, the increased adhesion is desirable in certain processes employing bacteria, as is for example the wastewater treatment. This thesis reviews main theories used for prediction of bacterial adhesion, describes the process of adhesion and physical, chemical and biological factors that influence it. It also contains an overview of antiadhesive and antibacterial surfaces. Key words: Bacteria, adhesion, pili, flagellum, antiadhesive and...
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Stability of protein complexes in the cytoskeleton of the eukaryotic flagellum
Pružincová, Martina ; Varga, Vladimír (advisor) ; Čajánek, Lukáš (referee)
The cilium/flagellum is a complex organelle protruding from the cell body and functioning in motility, sensing, and signalling. It is composed of hundreds of protein constituents, the majority of which comprise the flagellar cytoskeleton - the microtubule-based axoneme. Because the flagellum lacks ribosomes, its protein constituents have to be imported from the cell body and delivered to proper locations. Moreover, these proteins have to retain their function over a considerable length of time, despite the mechanical stress caused by flagellar beating and due to environmental exposure. This raises the question whether and where protein turnover occurs. Previously, it was established that Chlamydomonas reinhardtii flagella are dynamic structures (Marshall & Rosenbaum, 2001). In contrast, in the Trypanosoma brucei flagellum axonemal proteins are remarkably stable (Vincensini et al., 2018). However, the questions of axonemal assembly and stability were so far investigated only for a small number of proteins and during relatively short periods. Moreover, in these experiments expression of studied proteins was controlled by non-native regulatory elements. To elucidate the site of incorporation of proteins from all major axonemal complexes and to find out if and where the protein turnover occurs, T....
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Monoxenous trypanosomatids of invertebrates
Havlová, Jolana ; Votýpka, Jan (advisor) ; Varga, Vladimír (referee)
The class Kinetoplastea contains free-living and parasitic species. One of the most dominant group within the class is the order Trypanosomatida which includes obligate parasites (Trypanosoma, Leishmania) infecting a wide range of hosts. Some species are serious pathogens of humans and domestic animals and cause considerable losses. However, the majority of trypanosomatids belongs to monoxenous parasites of insect which are usually harmless to their hosts. Monoxenous trypanosomatids predominantly infect Hemiptera and Diptera. This diploma thesis is focused on the detection of monoxenous trypanosomatids in cockroaches captured in the Czech Republic and cockroaches from different breedings. Cockroaches are very suitable mechanical vectors of many different pathogens (including parasites) and are significant health threat for humans and animals. First trypanosomatids in cockroaches were documented at the beginning of the 20th century, but there is no study focused on this topic specifically. Another aim of this thesis is morphological and ultramicroscopic analysis and the study of the host specificity of the recently described species Herpetomonas tarakana, isolated from a cockroach. My findings were partly used in the already published study "Diversity of trypanosomatids in cockroaches and the...
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Isolation of bacterial flagellin as an immunogen for the preparation of antibodies for cystic fibrosis patients
Supová, Veronika ; Hodek, Petr (advisor) ; Kubíčková, Božena (referee)
Cystic fibrosis (CF) is an autosomal recessive disease caused by CFTR gene mutation. The consequence of this mutation is an improper function of the chloride channel consisting of CFTR protein. Due to this abnormality, the transport of chloride ions is either reduced or inhibited completely, which leads to a mucus secretion. Mucus is mostly created in the lungs and it is the ideal environment for pathogenic bacteria like Pseudomonas aeruginosa (PA). Pseudomonas aeruginosa is an aerobic, gram-negative conditioned pathogen occurring in patients with weakened immunity such as patients with CF, who are often hospitalized. PA has one polar flagellum which contains filament composed of a protein called flagellin. The flagellum is one of the most important virulence factors of PA bacteria. This thesis focused on the isolation of flagellin from Pseudomonas aeruginosa flagella. The isolated flagellin will serve as an antigen for the preparation of prophylactic antibodies for CF patients. The isolation of flagellin was carried out by four methods using combinations of precipitation and centrifugation. The published methods were optimized to achieve the isolation of flagellin. Final samples and intermediates were analyzed by SDS-electrophoresis on polyacrylamide gel and the presence of flagellin was verified...
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Sperm centrioles and their role in reproduction
Vlčková, Monika ; Frolíková, Michaela (advisor) ; Liška, František (referee)
Centrioles are evolutionarily conserved protein structures composed of microtubules. In somatic cells, centrioles serve as the basal body of cilia and flagella and allow the assembly of pericentriolar material, thereby creating the centrosome. Without centrosome, animal cells are not capable of nuclear division. Centrioles do not arise de novo and their formation always requires the presence of a preexisting centriole. Since there are no centrioles in the egg at the time of fertilization, unlike spermatozoa, sperm is the carrier of centrioles and therefore all of the centrioles of the emerging organism are of paternal origin. There are two centrioles in the sperm - a cylindrical shape proximal and a distal one, which is perpendicular to the proximal. The sperm centrioles are the basis for the formation of the sperm flagellum and after fertilization form the mitotic spindle of the zygote, necessary for equal DNA and cell distribution. It follows from the above that the presence of centrioles in sperm is essential in mammals and defects in their structure may lead to male sterility or embryo development disorders. However, sperm centrioles differ from somatic centrioles in their structure and behavior and understanding these differences is one of the important tasks of reproductive biology.
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Stability of protein complexes in the cytoskeleton of the eukaryotic flagellum
Pružincová, Martina ; Varga, Vladimír (advisor) ; Čajánek, Lukáš (referee)
The cilium/flagellum is a complex organelle protruding from the cell body and functioning in motility, sensing, and signalling. It is composed of hundreds of protein constituents, the majority of which comprise the flagellar cytoskeleton - the microtubule-based axoneme. Because the flagellum lacks ribosomes, its protein constituents have to be imported from the cell body and delivered to proper locations. Moreover, these proteins have to retain their function over a considerable length of time, despite the mechanical stress caused by flagellar beating and due to environmental exposure. This raises the question whether and where protein turnover occurs. Previously, it was established that Chlamydomonas reinhardtii flagella are dynamic structures (Marshall & Rosenbaum, 2001). In contrast, in the Trypanosoma brucei flagellum axonemal proteins are remarkably stable (Vincensini et al., 2018). However, the questions of axonemal assembly and stability were so far investigated only for a small number of proteins and during relatively short periods. Moreover, in these experiments expression of studied proteins was controlled by non-native regulatory elements. To elucidate the site of incorporation of proteins from all major axonemal complexes and to find out if and where the protein turnover occurs, T....
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Monoxenous trypanosomatids of invertebrates
Havlová, Jolana ; Votýpka, Jan (advisor) ; Varga, Vladimír (referee)
The class Kinetoplastea contains free-living and parasitic species. One of the most dominant group within the class is the order Trypanosomatida which includes obligate parasites (Trypanosoma, Leishmania) infecting a wide range of hosts. Some species are serious pathogens of humans and domestic animals and cause considerable losses. However, the majority of trypanosomatids belongs to monoxenous parasites of insect which are usually harmless to their hosts. Monoxenous trypanosomatids predominantly infect Hemiptera and Diptera. This diploma thesis is focused on the detection of monoxenous trypanosomatids in cockroaches captured in the Czech Republic and cockroaches from different breedings. Cockroaches are very suitable mechanical vectors of many different pathogens (including parasites) and are significant health threat for humans and animals. First trypanosomatids in cockroaches were documented at the beginning of the 20th century, but there is no study focused on this topic specifically. Another aim of this thesis is morphological and ultramicroscopic analysis and the study of the host specificity of the recently described species Herpetomonas tarakana, isolated from a cockroach. My findings were partly used in the already published study "Diversity of trypanosomatids in cockroaches and the...
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Bacterial adhesion to surfaces
Budil, Jakub ; Beranová, Jana (advisor) ; Kuthan, Martin (referee)
Bacterial adhesion is a process of attachment of a planktonic cell to the surface and the necessary step for further biofilm formation. Most bacterial species are capable of biofilm formation. Bacteria within biofilm exhibit increased resistance to antibiotics and other external factors, which makes the eradication of adhered bacteria rather difficult. Bacterial adhesion is very complex process affected by many physical and chemical factors as well as by the adhesive molecules present on the substrate and cell surface. Accurate description of these factors enables to design appropriate modifications of both the substrate and the environment with the aim to fabricate the antiadhesive and antibacterial surfaces or to increase the efficiency in bacteria-based biotechnological processes. Whereas in medical applications the main goal is to decrease bacterial adhesion, the increased adhesion is desirable in certain processes employing bacteria, as is for example the wastewater treatment. This thesis reviews main theories used for prediction of bacterial adhesion, describes the process of adhesion and physical, chemical and biological factors that influence it. It also contains an overview of antiadhesive and antibacterial surfaces. Key words: Bacteria, adhesion, pili, flagellum, antiadhesive and...
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The Demembranation as a Tool to Study a Movement and Physiology of Activation of Fish Spermatozoa
BLAŽKOVÁ, Jaroslava
The aim of the thesis was the summarization of information about the demembranation method as a means of study of the physiology of activation and motility of fish spermatozoa. A simple demonstration of the method is also presented. The demembranation of fish sperm is a method, in which cytoplasmic membrane (the phospholipid bilayer, which separate intercellular and extracellular environments) is destroyed. A mild non-ionic detergent, TrintonX-100, was used in the study. Common carp (Cyprinus carpio) as a representative member of Teleost fish and sterlet as a member of Chondrostean (elasmobranch) fish were studied in the thesis.
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