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Encystation and life cycle of free living amoebae of the genus Acanthamoeba spp.
Bínová, Eva ; Nohýnková, Eva (advisor) ; Dyková, Iva (referee) ; Ondriska, František (referee)
Amoebae of the genus Acanthamoeba spp. are free-living unicellular organisms found in disparate ecosystems all over the world. Due to their ability to invade human body, evade its defensive mechanisms and cause extensive tissue damage, Acanthamoeba infection can lead to serious, if rare, diseases, affecting most commonly the eye and the central nervous system. Specific therapy for Acanthamoeba infections is not available. A major reason for therapeutic failure in ameobiasis is the ability of the protist to differentiate into resistant stages. These are cysts, known to be formed under prolonged unfavorable conditions, both in the environment and the infected tissues, and the pseudocysts, less durable but rapidly formed under acute stress. The present thesis focuses on as yet unexplored mechanisms of resistance of cysts and pseudocysts. Moreover, further characteristics distinguishing cysts and pseudocysts as well as the processes involved in their formation are investigated. One of the issues addressed is a presence of protective carbohydrate compounds mannitol and trehalose that participate in defensive reactions against abiotic stress in many organisms. Although putative genes for enzymes of the trehalose and mannitol synthetic pathways are present in the genome of Acanthamoeba, only one of the...
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Live cycle of the free-living amoeba. Differentiation of amoebae of the genera Acanthamoeba and Balamuthia
Klieščiková, Jarmila ; Nohýnková, Eva (advisor) ; Ondriska, František (referee) ; Walochnik, Julia (referee)
Free-living pathogenic amoebae Acanthamoeba spp. and Balamuthia mandrillaris are causative agents of important diseases of human: rarely occurring but highly fatal granulomatous amoebic encephalitis (both) and keratitis (Acanthamoeba). One of the reasons for the problematic therapy is differentiation into highly resistant cysts often found in affected tissues. In our study we have found that correct encystation in Acanthamoeba requires apart from others, the presence of functioning Golgi apparatus transporting the cyst wall material to the cell surface; glycogen phosphorylase degrading glycogen into glucose which seems to be further used for cellulose synthesis and two non-constitutive cellulose synthases. Acanthamoeba cellulose synthases seem not to be inhibited by known herbicides. In the cyst wall of acanthamoebae we detected cellulose, -mannan, and -1, 3-1, 4-linked glucan [lichenin or mixed-linkage glucan (MLG)]. Cellulose is present in the inner (endocyst) and the outer (exocyst) layers of the cyst wall, whereas-mannan and MLG are found in the endocyst. In a protozoan organism, MLG was detected for the first time. The MLG of Acanthamoeba has a similar composition to that found in barley with high amount of cellobiosyl and cellotriosyl followed by cellotetraosyl units. In contrast, with...
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