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The role of humoral factors in the snail immune response against schistosomes
Košťáková, Monika ; Horák, Petr (advisor) ; Dvořák, Jiří (referee)
Digenetic trematodes such as Schistosoma mansoni use molluscs, mainly Gastropoda in their life cycle, as their intermediary hosts. e internal defense system (IDS) of snails is composed of immune cells called hemocytes, which are the main effectors and act jointly with soluble components. Humoral factors could in uence directly the parasite's larval stage, the activity of hemocytes and also may serve in recognition of the parasite. Lectins are considered to be the main component of humoral immunity. ey have a primary role in non-self recognition. Other protein group with lectin-like activity called FRePs was found in Biomphalaria glabrata. eir unique structure contains a brinogen and an immunoglobulin-like domain. Cytokine-like molecules may play very important role in defense as well. Many molecules are present in hemolymph and their levels change during infection. e response to parasitosis is therefore very complex and still awaits further clari cation.
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Inhibitors of proteolytic enzymes of Trematodes
Šteiger, Vladimír ; Kašný, Martin (advisor) ; Salát, Jiří (referee)
i Abstract Trematodes are important parasites possessing various localization in the bodies of invertebrate and vertebrate hosts, including human; therefore they are subject of long time intensive worldwide research. Trematodes developed various adaptations and strategies (some of them have also molecular background) enable them to survive in the host bodies. Trematodes produce large amount of different molecules, which are involved in various physiological processes. Inhibitors of proteolytic enzymes form a large group of biologically active compounds, e.g. they regulate the activity of peptidases or modulate host immune response. Many of these inhibitors are investigated as potential candidates in chemotherapeutic fight against trematodes. This thesis reviews the information concerning the natural inhibitors produced by trematodes and also synthetic inhibitors. Key words: Inhibitor, trematode, peptidase, serpin, cystatin i
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Glycocalyx shedding by cercariae of bird schistosomes
Chaloupecká, Jana ; Mikeš, Libor (advisor) ; Štěrba, Ján (referee)
Trichobilharzia spp. are avian schistosomes related to medically important human parasites of the genus Schistosoma. Penetrating cercariae are well known as causative agent of cercarial dermatitis in humans. Cercariae actively penetrate the skin of definitive hosts and transform into schistosomula. This process is preceded by cercarial tail detachment and includes emptying of penetration glands and extensive surface changes. One of these changes is the loss of highly immunogenic glycocalyx which represents a protective coat in the aquatic environment. The glycocalyx has specific composition of saccharide molecules which are bound to lipids or proteins on the membrane of cercarial tegument. There is only limited information about the mechanism of shedding. Hypotheses based on indirect evidences suggest that peptidases or (phospho)lipases from penetration glands could be involved. This work describes the changes in surface glycosylation during transformation of cercariae into schistosomula by fluorescently labelled lectins and monoclonal antibodies against Lewis X antigen. Lectins UEA-I, LTA and PNA have been chosen as markers of transformation of T. regenti. Further, our experiments have been focused on shedding of cercarial glycocalyx. During in vitro induction of penetration gland emptying and...
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Features and functions of glycocalyx of trematode cercariae
Chaloupecká, Jana ; Mikeš, Libor (advisor) ; Kašný, Martin (referee)
Trematodes are parasites from phylum Platyhelminthes which have compex life cycles involving two to four hosts. This work focuses especially on trematodes of the family Schistosomatidae. Their cercariae which leave the snail intermediate host, actively penetrate the skin of definitive hosts and transform into schistosomula. This is accompanied by detachment of cercarial tail and emptying of penetration glands. During transformation, cercarial bodies undergo extensive ultrastructural and molecular changes. One of these changes is the loss of surface glycocalyx which represents a protective coat in the aquatic environment. In glycocalyx shedding, participation of proteolytic enzymes from cercarial penetration glands is expected during invasion of the host. Glycocalyx has specific composition of saccharide molecules which are bound to lipids or proteins on the membrane of cercarial tegument. This work describes the origin, ultrastructure, saccharide composition, function and shedding mechanism of cercarial glycocalyx.
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Cathepsins L of Diplostomum pseudospathaceum cercariae
Perháčová, Terézia ; Mikeš, Libor (advisor) ; Hartmann, David (referee)
This study is focused on cercarial cysteine peptidases of the trematode Diplostomum pseudospathaceum. It follows previous research which confirmed the presence of a 24kDa cysteine peptidase in cercariae biochemically and by mass spectrometry. It was postulated, that the function of this peptidase is histolytic, when cercariae penetrate the tissues. During an attempt to purify this peptidase and characterize its peptidolytic activity, it was found out that the cercarial homogenate containsmore different peptidases varying in their pI. Tests of peptidolytic activity and inhibition have shown that these peptidases are cathepsin L-like. They are active over a broad spectrum of pH with optima of activities in weakly acidicor neutral pH. Using degenerate primers based on conserved motifs of cysteine pepridases, partial sequences of three genes for cathepsin L of D. pseudospataceum (DpCL1, 2 a 3) were obtained. Then the complete sequences of DpCL2 and 3 genes and partial sequence (without 5'end) of DpCL1 were obtained by RACE PCR. To confirm function of these peptidases we tried to immunolocalize them. We assumed that they are localized in penetration glands. Preliminary results suggested that some of the cathepsins could be also localized in the gut of cercariae. For more detailed biochemical...
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Aporocotyle simplex fluke from the flounder Hippoglossoides platessoides in Svalbard.
HÁJKOVÁ, Šárka
In my bachelor thesis I studied trematodes from plaice Hippoglossoides platessoides obtained during expeditions to Svalbard in 2014 and 2018. My goal was morphological and molecular processing of the obtained material, which allowed me to confirm the identification of the flukes as Aporocotyle simplex Odhner, 1900. Aporocotyle simplex is a common parasite of plaice, ocurring mainly in Hippoglossoides platessoides, but also in Limanda limanda and Pleuronectes platessa. The species A. simplex is geographically distributed in the Gulf of St. Lawrence in Canada, off the west coast of Sweden, in the Barents Sea and Bering Sea, off the coast of Kamchatka and Greenland. My results showed that the species A. simplex is widespread in Svalbard. In the practical part, I dealt with methods that helped me to identify the studied specimens. I observed the trematode tegumental surface with aid of a scanning electrom microscope (JEOL JSM-7401F). The trematode internal organs were stained with Mayer-Schuberg carmine, which allowed me to observe and draw the mounted specimens using a light microscope (Olympus BX51) equipped with a drawing attachment. Furthermore, for molecular analysis, the DNA was isolated using the commercial Exgene Tissue SV mini kit (GeneAll). The isolated DNA (the gene for the large ribosomal subunit, 28S rRNA) was amplified by PCR, the obtained PCR products were sequenced on an automated sequencer (ABI Prism 3130xl or 3730xl, by SEQme). The newly acquired sequences (three in total) were aligned in the program Geneious 8.0.5with sequences available from GenBank (64 sequences). Phylogenetic analysis of the species relationships within the family Aporocotylidae was performed using the Maximum Likelihood method with TPM2u + F + G4 used as the best model. Aporocotyle simplex falls into a separate branch together with A. michaudi, from which however, it differs substantially by its morphology (distribution of tegumental spines on body and their number in the clusters, number of testes, ratio of esophagus length to body length) and geographical distribution (A. michaudi occurs in the South Atlantic Ocean).
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Inhibitors of proteolytic enzymes of Trematodes
Šteiger, Vladimír ; Kašný, Martin (advisor) ; Salát, Jiří (referee)
i Abstract Trematodes are important parasites possessing various localization in the bodies of invertebrate and vertebrate hosts, including human; therefore they are subject of long time intensive worldwide research. Trematodes developed various adaptations and strategies (some of them have also molecular background) enable them to survive in the host bodies. Trematodes produce large amount of different molecules, which are involved in various physiological processes. Inhibitors of proteolytic enzymes form a large group of biologically active compounds, e.g. they regulate the activity of peptidases or modulate host immune response. Many of these inhibitors are investigated as potential candidates in chemotherapeutic fight against trematodes. This thesis reviews the information concerning the natural inhibitors produced by trematodes and also synthetic inhibitors. Key words: Inhibitor, trematode, peptidase, serpin, cystatin i
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Cathepsins L of Diplostomum pseudospathaceum cercariae
Perháčová, Terézia ; Mikeš, Libor (advisor) ; Hartmann, David (referee)
This study is focused on cercarial cysteine peptidases of the trematode Diplostomum pseudospathaceum. It follows previous research which confirmed the presence of a 24kDa cysteine peptidase in cercariae biochemically and by mass spectrometry. It was postulated, that the function of this peptidase is histolytic, when cercariae penetrate the tissues. During an attempt to purify this peptidase and characterize its peptidolytic activity, it was found out that the cercarial homogenate containsmore different peptidases varying in their pI. Tests of peptidolytic activity and inhibition have shown that these peptidases are cathepsin L-like. They are active over a broad spectrum of pH with optima of activities in weakly acidicor neutral pH. Using degenerate primers based on conserved motifs of cysteine pepridases, partial sequences of three genes for cathepsin L of D. pseudospataceum (DpCL1, 2 a 3) were obtained. Then the complete sequences of DpCL2 and 3 genes and partial sequence (without 5'end) of DpCL1 were obtained by RACE PCR. To confirm function of these peptidases we tried to immunolocalize them. We assumed that they are localized in penetration glands. Preliminary results suggested that some of the cathepsins could be also localized in the gut of cercariae. For more detailed biochemical...
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Orientation of helminths during host seeking
Vojtová, Terezie ; Mikeš, Libor (advisor) ; Horák, Petr (referee)
The parasitic helminths are often forced in the environment to search for their host quickly in order to continue the ontogenetic development, or else to finish it. To search for and indentify the host, they use various types of receptors which allow them to provoke a taxes corresponding to the oncoming stimulus and to react regarding the current situation. In respect to the concept relevant to the helminths, this work concerns trematodes, whose freely movable stadia are miracidium and cercacia, and then monogeneans which are viparous or oviparous with oncomiracidium larva, and nematodes during which the third larva stage moves. The representatives of the groups of Myxozoans and Ciliophorea are also mentionned, since they use similar mechanisms to look for hosts as helminths. This thesis includes the summary of the existing knowledge related to the orientation of helminths in the environment. On the basis of the data that is already known, it is possible to compare which mechanisms are used for host-seeking by particular species, and according to which signals from the surroundings they most often orientate themselves. Powered by TCPDF (www.tcpdf.org)
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Orientation of helminths during host seeking
Vojtová, Terezie ; Mikeš, Libor (advisor) ; Horák, Petr (referee)
Helminths are often forced in the environment to search for their host quickly in order to continue the ontogenetic development, or else to finish it. To search for and indentify the host, they use various types of receptors which allow them to provoke a taxes corresponding to the oncoming stimulus and to react to the presence of the host. This thesis includes the summary of the existing knowledge related to the orientation of helminths, the comparison of the mechanisms used by the searching of a host and signals in the environment. In respect to the concept relevant to the helminths, this work concerns trematodes, whose freely movable stadia are miracidium and cercaria, and then monogeneans which are viparous or oviparous with oncomiracidium larva, and nematodes with the third infective larva stage. The representatives of fish parasites of Myxozoans and Ciliophorea are also mentionned, since they use similar mechanisms to look for hosts as helminths. Powered by TCPDF (www.tcpdf.org)
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