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Biogenesis and function of peroxisomes in parasitic protists
Nguyen, Thu Duong ; Tachezy, Jan (advisor) ; Verner, Zdeněk (referee)
Peroxisomes are found in majority of eukaryotes and possibly originate from the last eukaryotic common ancestor, as the proteins involved in their biogenesis are highly conserved. Enzymatic content of peroxisomes is highly diverse and their metabolic activity may vary depending on the cell type. Metabolically, peroxisomes interact with mitochondria and these organelles are functionally linked. Peroxisomes in unicellular eukaryotes are very diverse. The peroxisomes in kinetoplastids are named glycosomes because they contain several glycolic enzymes. The loss of peroxisomes occurs in certain representatives of Alveolata, and peroxisomes are entirely absent in Metamonada group. Anaerobic peroxisomes were found in Archamoebidae. They are metabolically quite distinct from their aerobic counterparts. The properties of these peroxisomes are related to the lifestyle of these protists. This work briefly summarizes information regarding the biogenesis of peroxisomes and their functions, their functional connection with mitochondria and then provide characteristics of peroxisomes in selected protists.
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Double expression systems with respect to parasitic protozoa
Gromyko, Anastasiia ; Verner, Zdeněk (advisor) ; Kraeva, Natalia (referee)
Protozoan parasite infections continue to pose a significant health challenge in developing countries, resulting in hundreds of thousands of deaths each year. These parasites exhibit a complex multi-stage life cycle and possess unique cellular structures. However, many of their biological processes remain poorly understood. Multigene expression is a promising approach to address this knowledge gap, as it enables the expression of functional protein complexes in vivo, the addition of fluorescent protein tags for visualization of protein localization within the cell, and the study of protein-protein interactions. This bachelor's thesis reviews the current knowledge on available systems and approaches for studying key model parasitic protozoan species. Keywords: expression systems, Trypanosoma brucei, Leishmania tarentolae, Entamoeba histolytica, Giardia intestinalis, Trichomonas vaginalis, Toxoplasma gondii, Plasmodium falciparum, pET-Duet, CRISPR-Cas9
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Pentoses as energy sources of Mastigamoeba balamuthi
Novotná, Monika ; Verner, Zdeněk (advisor) ; Čepička, Ivan (referee)
Mastigamoeba balamuthi is a free-living amoeba, a sister group to one of the most widespread parasitic agent Entamoeba histolytica. During its lifetime, M. balamuthi can utilize at least four different energy sources to produce ATP. In addition to glucose, a standard cultivation energy source, the amoeba can utilize pentoses xylose and arabinose, as well as an amino acid L-arginine. In this work, energy source preference was assessed. Morphological and physiological characteristics including cell growth and size, motility, number of nuclei and metabolic end products were evaluated. It was concluded that glucose appears to be a less preferred energy source than pentose upon evaluating statistically significant differences. Therefore, it was proposed to replace the commonly used hexose with the preferred xylose in the culture medium of M. balamuthi.
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Modes of resolution of a debtor's insolvency
Verner, Zdeněk ; Pohl, Tomáš (advisor) ; Macková, Alena (referee)
The methods of solving the debtor' s insolvency using the "sanitation" principle It has been 5 years since the Czech Insolvency Act was enacted. Therefore is it a good opportunity to summarize the pros and cons of the current situation in this branch of law. The Insolvency Act introduced the concept of two approaches to dealing with the debtor' s insolvency which are new in the Czech legal system: the restructuring and the discharge. Both of them share the "sanitation" principle which enables the debtor to continue their existence after having adopted certain measures. The thesis consists of five major chapters. First section provides the basic explanation of the concept of insolvency and the reasons for the special treatment for the bankruptcy of those who have more creditors. There is also a brief description of the insolvency' s procedure. The second chapter deals with the restructuring, which is the method of solving the insolvency of big companies (entrepreneurs). The third chapter focuses on discharge which is intended for those who are not engaged in business activities (non-entrepreneurs) and which enables them to get rid of their debts. The discharge can be performed in two modes: either by the liquidation of the assets or through the performance of the payment calendar. The chapter four and...
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The role of mitochondria in the pathogenesis
Marková, Lenka ; Doležal, Pavel (advisor) ; Verner, Zdeněk (referee)
This thesis is focused on the existing data referring to role of mitochondria in the pathogenesis of several important diseases. It introduces mitochondrial proteins, their role and metabolism, dysfunction of which is behind pathogenetic processes. The thesis also summarizes possible mitochondrial damage, its progress and consequences, which can lead to diseases or aggravate their process. It also pays attention to the role of mitochondria during oncogenesis as well as important neurodegenerative disorders such as Alzheimer's or Parkinson's disease, Charcot-Marie-Tooth disease, amyotrophic lateral sclerosis or autosomal dominant optic atrophy. The thesis mentions the role of disrupted mitochondrial dynamics in type 2 Diabetes. In conclusion the thesis mentions the role of mitochondria and their damage in relation to infection by the parasites Toxoplasma gondii and Trypanosoma cruzi.
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Functional study of the SUF pathway in the cell of Monocercomonoides exilis and Paratrimastix pyriformis
Zelená, Marie ; Hampl, Vladimír (advisor) ; Verner, Zdeněk (referee)
The synthesis of iron-sulfur clusters is an essential cellular process, which depends on complex biosynthetic pathways. In model eukaryotes, these pathways are the ISC pathway in the mitochondria and the CIA pathway in the cytosol. A recent genome and transcriptome analysis showed, that an amitochondriate protist Monocercomonoides exilis lacks the canonical ISC pathway, which has been replaced by a bacterial SUF pathway. A close free-living relative of M. exilis, Paratrimastix pyriformis possesses a mitochondrion-related organelle, yet also possesses a SUF pathway instead of ISC. The acquisition of the SUF pathway has been suggested as the primordial cause for mitochondrial loss in M. exilis, which is the first documented eukaryotic organism without a mitochondrion. The SUF pathway has been the subject of numerous studies in bacteria, however, its role as the core provider of iron-sulfur clusters for eukaryotic cells has been reported in merely a handful of eukaryotes and was based predominantly on genomic data. This thesis focuses on the putative ATPase SufC and the putative scaffold protein SufB. Both proteins were successfully produced in recombinant forms. SufC has been found to possess ATPase activity in vitro, which was increased upon interaction with SufB. The conditions for theATPase...
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Dynamics of Giardia intestinalis encystation.
Vinopalová, Martina ; Doležal, Pavel (advisor) ; Verner, Zdeněk (referee)
Giardia intestinalis is an anaerobic parasite, that colonizes the small intestine of humans and other vertebrate hosts. This cosmopolitan parasite, which causes diarrhoea, is transmitted by contaminated water or food via a resistant stage, the cyst. The encystation process involves a number of events that lead to a complete reconstruction of the cell into the form of infectious cyst. The aim of this work was to visualize these modifications in vivo by means of enzymatic labelling of proteins. For the purposes of this work, enzymatic tags Y-FAST and HaloTag were chosen, as they enable visualizing live cells under anaerobic conditions. Chimeric protein constructs were created to visualize the dynamics of the encystation vesicles, the structures of endoplasmic reticulum, the adhesive disc and mitosis. Using the developed constructs, we successfully followed the dynamics of the encystation vesicles and the adhesive disc in vivo. Finally, this work has provided novel molecular tools, which will be used to follow the overall redesign of the parasite cell during encystation.
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The role of mitochondria in the pathogenesis
Marková, Lenka ; Doležal, Pavel (advisor) ; Verner, Zdeněk (referee)
This thesis is focused on the existing data referring to role of mitochondria in the pathogenesis of several important diseases. It introduces mitochondrial proteins, their role and metabolism, dysfunction of which is behind pathogenetic processes. The thesis also summarizes possible mitochondrial damage, its progress and consequences, which can lead to diseases or aggravate their process. It also pays attention to the role of mitochondria during oncogenesis as well as important neurodegenerative disorders such as Alzheimer's or Parkinson's disease, Charcot-Marie-Tooth disease, amyotrophic lateral sclerosis or autosomal dominant optic atrophy. The thesis mentions the role of disrupted mitochondrial dynamics in type 2 Diabetes. In conclusion the thesis mentions the role of mitochondria and their damage in relation to infection by the parasites Toxoplasma gondii and Trypanosoma cruzi.
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Modes of resolution of a debtor's insolvency
Verner, Zdeněk ; Pohl, Tomáš (advisor) ; Macková, Alena (referee)
The methods of solving the debtor' s insolvency using the "sanitation" principle It has been 5 years since the Czech Insolvency Act was enacted. Therefore is it a good opportunity to summarize the pros and cons of the current situation in this branch of law. The Insolvency Act introduced the concept of two approaches to dealing with the debtor' s insolvency which are new in the Czech legal system: the restructuring and the discharge. Both of them share the "sanitation" principle which enables the debtor to continue their existence after having adopted certain measures. The thesis consists of five major chapters. First section provides the basic explanation of the concept of insolvency and the reasons for the special treatment for the bankruptcy of those who have more creditors. There is also a brief description of the insolvency' s procedure. The second chapter deals with the restructuring, which is the method of solving the insolvency of big companies (entrepreneurs). The third chapter focuses on discharge which is intended for those who are not engaged in business activities (non-entrepreneurs) and which enables them to get rid of their debts. The discharge can be performed in two modes: either by the liquidation of the assets or through the performance of the payment calendar. The chapter four and...
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