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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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How Intracellular Pathogens Manipulate Cellular Trafficking.
Petrů, Markéta ; Doležal, Pavel (advisor) ; Pyrih, Jan (referee)
Many intracellular single-celled organisms belong to medically important human pathogens. The selected parasites are subject of this thesis - Chlamydia spp., Legionella pneumophila, Trypanosoma cruzi and Toxoplasma gondii - as well as their interactions with the vesicular transport of the host cell. Basic pathways of vesicular transport are delineated and important participating molecules described. Furthermore, the effector proteins of pathogens that interact with these molecules are included. The special chapter is devoted to phenomenon of mimetics of SNARE proteins by bacteria. The manuscript concludes with a chapter on LpSNARE of Legionella pneumophila, which was found in our laboratory and which is a topic of my experimental work herein.
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The role of Toxoplasma gondii on the expression of Major Urinary Proteins in the house mouse
Hladovcová, Denisa ; Stopka, Pavel (advisor) ; Hrdý, Ivan (referee)
Major Urinary Proteins (MUP) are pheromonal transmitters involved in chemical communication in rodents. Complexes of MUPs and ligands mediate information about genetical background of an individual and co-create individual scent profile. They play a significant role in kinship determination, the crucial factor in the choice of a mating partner. It is assumed that the MUP production is energetically demanding due to the pressure of sexual selection, and the transmitted signal is thus supposed to be honest. This theory hasn't yet been experimentally tested, neither was proven the effect of another intense selection pressure- a parasitic infection. In my thesis, I describe the effect of an infection of a parasite Toxoplasma gondii on MUP production. The results suggest that the production is altered in both sexes, the production decreasing in males to the level of male castrates (or the female level). Considering the results of more detailed analysis of the infection we assume that the energetical demands of the MUP production doesn't allow the production of ordinary amounts if MUPs during the first phase of the infection as the energy is primarily devoted to the infection control. The increased production in latter phase can be attributed to the importance of scent communication. Decreased levels of...
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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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Sexual transmission of Toxoplasma gondii from males to females: experimental verification using laboratory animal model
Navrátil, Jiří ; Kodym, Petr (advisor) ; Votýpka, Jan (referee)
Toxoplasma gondii is cosmopolitly living parasite which prevalence in human extends to tens of percent. In its life cycle it uses any homoiothermic vertebrate as an intermediate host. The definitive host are felines from Felidae family. The acute phase of infection is medically important in immunocompromised pacients and by its risk of congenital toxoplasmosis in pregnant women who never suffered from this illness before. Infection could have serious and rarely even lethal consequences in both cases. This thesis focuses on experimental verification of theory of sexual transmission of toxoplasmosis from male to female on laboratory mice. Possible transmission was tested in acute phase and latent phase of infection. The result was negative in both cases. Moreover, we observed the parasite's affinity to tissue of organs in male mice by PCR technique. Particularly, our interest was in comparing genital organs with others. It was discovered that lungs and spleen are the most infected organs in acute phase of infection. Toxoplasma was also present in genital organs (especially in epididymis) but not more frequently than in others. We observed statistically significant difference between sexual and non-sexual organs in acute and latent toxoplasmosis - non-sexual organs were more infected in both phases....
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Sexual transmission of Toxoplasma gondii from males to females: experimental verification using laboratory animal model
Navrátil, Jiří ; Kodym, Petr (advisor) ; Votýpka, Jan (referee)
Toxoplasma gondii is cosmopolitly living parasite which prevalence in human extends to tens of percent. In its life cycle it uses any homoiothermic vertebrate as an intermediate host. The definitive host are felines from Felidae family. The acute phase of infection is medically important in immunocompromised pacients and by its risk of congenital toxoplasmosis in pregnant women who never suffered from this illness before. Infection could have serious and rarely even lethal consequences in both cases. This thesis focuses on experimental verification of theory of sexual transmission of toxoplasmosis from male to female on laboratory mice. Possible transmission was tested in acute phase and latent phase of infection. The result was negative in both cases. Moreover, we observed the parasite's affinity to tissue of organs in male mice by PCR technique. Particularly, our interest was in comparing genital organs with others. It was discovered that lungs and spleen are the most infected organs in acute phase of infection. Toxoplasma was also present in genital organs (especially in epididymis) but not more frequently than in others. We observed statistically significant difference between sexual and non-sexual organs in acute and latent toxoplasmosis - non-sexual organs were more infected in both phases....
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Influence of latent toxoplasmosis on human fertility
Hlaváčová, Jana ; Kaňková, Šárka (advisor) ; Calda, Pavel (referee)
Toxoplasma gondii is one of the most widespread parasitic protozoa in humans in developed countries. It has a wide range of hosts including various warm-blooded animals and humans. Recent studies suggest that it could influence the fertility of humans. The aim of this work was to find out, what is the prevalence of toxoplasmosis in groups of women and men with fertility problems and whether Toxoplasma gondii has an effect on specific fertility parameters. In women, Significant positive correlation between toxoplasmosis and the percentage of oocytes in the follicles was found among women. Group of men with semen pathology diagnosis revealed a statistically significant negative association between toxoplasmosis and the percentage of morphologically normal sperm. It seems that latent toxoplasmosis in men can also worsen the negative effect of tobacco smoking on fertility. The impairment of fertility in men could be the result of the manipulation activity of Toxoplasma gondii, which leads to more frequent sexual intercourse due to decreased ability to conceive. This could increase the likelihood of sexual transmission, which is currently suggested as one of the possible ways of transmission of toxoplasmosis.
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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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The relation between latent toxoplasmosis and schizophrenia
Sglundová, Markéta ; Flegr, Jaroslav (advisor) ; Holub, David (referee)
The relation between the intracellular parasite Toxoplasma gondii and schizophrenia was studied and confirmed in several studies. There are proofs that latent toxoplasmosis deteriorates the course of schizophrenia. The goal of this study was to clarify whether there are differences between subjectively perceived quality of life of infected and non-infected psychiatric patients. We tested men and women from the PCP treated for the diagnosis of schizophrenia, who undertook serology blood tests for detection of anamnestic T. gondii antibodies. Patients also filled out two questionaires that assessed subjectively perceived quality of life - SOS-10 and WHOQOL-BREF. Patients filled out these questionaires twice, both at the start of the study before coming to psychiatric therapeutical institution and after one year. During the first testing were the questionaires filled out by 171 persons, after one year by 108 persons. Based on the results of these tests we found out that latent toxoplasmosis had a negative impact on subjectively perceived quality of life of schizophrenia patients, particularly on infected women. On the other hand, the quality of life of non-infected patients improved during one year. The effect of toxoplasmosis on quality of life was gender related, which could be explained by already...
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