Project description:Opisthorchis felineus is one of the three most medically important species belonging to the family of fish-borne zoonotic trematodes known as Opisthorchiidae. O. felineus is endemic to the river plains of Western Siberia and Eastern Europe and it is estimated that more than 1.6 million people could be infected with this parasite. To aid in the development of better control and diagnosis strategies for opisthorchiasis, we set out to deduce the secreted proteome of O. felineus. Adult flukes were collected from experimentally infected hamsters and cultured in vitro in serum-free media. We extracted proteins from different compartments of the O. felineus secretome, including (i) soluble excretory/secretory (ES) products; (ii) secreted microvesicles (MVs); and (iii) tegument. The tegument was further separated into three fractions of varying solubility via sequential extraction.
Project description:Opisthorchis felineus is one of the three most medically important species belonging to the family of fish-borne zoonotic trematodes known as Opisthorchiidae. O. felineus is endemic to the river plains of Western Siberia and Eastern Europe and it is estimated that more than 1.6 million people could be infected with this parasite. To aid in the development of better control and diagnosis strategies for opisthorchiasis, we set out to deduce the secreted proteome of O. felineus. Adult flukes were collected from experimentally infected hamsters and cultured in vitro in serum-free media. We extracted proteins from different compartments of the O. felineus secretome, including (i) soluble excretory/secretory (ES) products; (ii) secreted microvesicles (MVs); and (iii) tegument. The tegument was further separated into three fractions of varying solubility via sequential extraction.
Project description:A significant number of pathological conditions, accompanied by chronic non-healing wounds, demands searching for new modern therapeutic approaches. Well-documented ability of O. felineus to initiate extracellular matrix (ECM) remodeling and liver epithelium regeneration suggests that its bioactive molecules may stimulate skin wound healing processes. The aim of this study was to investigate the wound healing potential of the Opisthorchis felineus excretory-secretory and lysate proteins on a murine model. The following methods were used for the study: histological (wounded skin condition), immunohistochemical (ECM, neoangiogenesis, O. felineus GST and TPx proteins), gene expression analysis (inflammation, angiogenesis, ECM condition). O. felineus excretory-secretory product (ESP) and lysate proteins have revealed wound-healing potential, they: i) reduce inflammation levels, ii) modulate vascular response, iii) stimulate collagen deposition and dermal ECM remodeling. Additional proteomic analysis of adult O. felineus ESP and lysate samples was conducted. Proteomic analysis approach called GeLC-MS / MS was chosen to study of the excretory-secretory product and lysate proteins. This approach is based on one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), in-gel protein digestion with trypsin followed by liquid chromatography-tandem mass spectrometry. The SDS-PAGE step allow to removes hemozoin also as detergents, buffers and salts from the protein extract that may interfere with mass spectrometry analysis
Project description:Background: Treatment and morbidity control of schistosomiasis relies on a single drug, praziquantel. Therefore, there is a pressing need to explore the effects of PZQ on the parasites at the molecular level and to pursue alternative and/or or synergistic drugs against schistosomiasis. Methodology: We used a custom-designed Schistosoma mansoni oligo-microarray to explore the effects of a sublethal dose of praziquantel on S. mansoni adult worms’ gene expression. We used functional analysis of gene interaction networks to identify differentially expressed genes that are known targets of other drugs already tested in humans for diverse disease conditions. Omeprazole, a proton pump inhibitor drug that is widely prescribed, was tested in combination with praziquantel. We comparatively assessed the efficacy of praziquantel or omeprazole alone and in combination against S. mansoni adult worms in vitro over a period of 120 hours. Principal Findings: We identified sets of genes that were affected by praziquantel on both paired and unpaired females, however with opposite gene expression patterns (up-regulated in paired and down-regulated in unpaired females), indicating that the transcriptomics changes induced by praziquantel are heavily influenced by the mating status. We also identified genes that were similarly affected by praziquantel in males and females. The use of sublethal doses of praziquantel together with omeprazole showed a significantly enhanced worm mortality in vitro compared to praziquantel alone, thus evidencing a synergic effect. Conclusions: Functional analysis of gene interaction networks is an important approach to identify genes whose expression is affected by one drug and that are at the same time known targets of other drugs already tested in humans for diverse disease conditions, thus pointing the latter as possible synergic drugs. Combined treatment with omeprazol increases the efficacy of praziquantel against S. mansoni adult worms in vitro, and additional in vivo studies are warranted.