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Effect of albendazole on the activity of selected enzymes in tapeworm Hymenolepis diminuta
Krejzová, Andrea ; Vokřál, Ivan (advisor) ; Szotáková, Barbora (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Andrea Krejzová Supervisor: PharmDr. Ivan Vokřál, Ph.D. Title of diploma thesis: Effect of albendazole on the activity of selected enzymes in tapeworm Hymenolepis diminuta The efficacy of anthelmintics used to treat diseases caused by helminths is not always sufficient, and in some cases, we are directly facing resistance to these drugs. Helminths, including tapeworms, are able to defend against the toxic effect of anthelmintics using several mechanisms. Xenobiotic metabolizing enzymes and transport proteins belong to these mechanisms. When xenobiotic metabolizing enzymes are induced, the efficacy of therapy may be significantly reduced. The effect of xenobiotic metabolizing enzymes on the drug resistance development has been already described in number of helminths. In tapeworms this information is still missing. Main aim of this study was to determine effect of drug albendazole on the activity of selected xenobiotic metabolizing enzymes in rat tapeworm (Hymenolepis diminuta). Tapeworms were incubated with albendazole (1 μM and 10 μM) for 24 hours. Then activities of selected enzymes in cytosol-like, microsome-like and mitochondria-like fractions were determined. This study is focused on...
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Zavedení helminta jako modelového organismu do kultury a charakterizace imunitní odpovědi hostitele
HLOŽKOVÁ, Oldřiška
The main goals of the present thesis were (i) to introduce a model commensal tapeworm, Hymenolepis diminuta, to the culture and (ii) characterization of a mucosal immune response in the host gut. Primarily, I focused on the establishment of a culture of H. diminuta under laboratory conditions based on its natural life cycle - grain beetles (Tenebrio molitor) as an intermediate host and outbred Wistar rats as definitive hosts. Infected beetles were bred at 28°C and due to this specific condition we were able to get infectious larvae three weeks after the first infection. To meet the second aim of this study, I examined a production of IgA antibodies against H. diminuta in the intestine of definitive host in the fecal samples using an indirect ELISA method. First, I performed several experiments focused on the optimization of H. diminuta antigen suitable for detection of IgA antibodies against H. diminuta in ELISA. Subsequently, I performed two experiments for detection of IgA fluctuation during H. diminuta infection in the rat model system. The strong production of IgA antibodies started always in the patent period of infection (i.e., when adult tapeworm is developed). In summary, my results suggest that H. diminuta has a strong immunomodulatory effect on the host organism.
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In vitro cultivation of tapeworm Hymenolepis diminuta - 2
Jandura, Dominik ; Vokřál, Ivan (advisor) ; Raisová Stuchlíková, Lucie (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Dominik Jandura Supervisor: PharmDr. Ivan Vokřál, Ph.D. Title of diploma thesis: In vitro cultivation of tapeworm Hymenolepis diminuta - 2 Aim of this diploma thesis was to obtain cycticercoids of the rat tapeworm (Hymenolepis diminuta), excyst them and find out the conditions for the maximal in vitro incubation period. As the intermediate host mealworm beetle (Tenebrio molitor) infected by the rat feces containing tapeworm eggs was used. Excystment was done using L-cystein and sodium tauroglycocholate. Excysted larvae were cultured in vitro (37 řC, 5 % CO2) in RPMI 1640 medium enriched with other substances chosen according previously published methods. Mainly sheep, mouse or rat liver extracts eventually in combination with yeast extract and sheep bile were used. The effect of tested substances on the cultivation was evaluated by measuring of the tapeworm's growth. The best effect on the grow of the tapeworms was observed using medium containing serum, yeast extract and sheep liver extract where tapeworms achieved length of 1561 µm after 16 days of incubation. The further growth was limited by appearance of pathologic formations.
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Molekulárně-fylogenetická charakteristika izolátu Hymenolepis diminuta udržovaného v laboratorních podmínkáh
ŘEŽÁBKOVÁ, Lucie
The main goal of the present study was to confirm the possibility of cryptic diversity within Hymenolepis diminuta group of tapeworms. We used molecular-fylogenetic analyses for characterization of the intraspecific genetic diversity of five isolates of Hymenolepis diminuta. Further, we wanted to compare our isolate of H. diminuta that is tested for purposes of the helminth therapy with other isolates obtained from collaborative institutions. Our results showed no cryptic diversity within H. diminuta clade of the five isolates and confirmed that our isolate is really H. diminuta previously characterized only based on its morphology.
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