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:BackgroundThere is increasing interest in the microbiome of the hepatobiliary system. This study investigated the influence of infection with the fish-borne liver fluke, Opisthorchis felineus on the biliary microbiome of residents of the Tomsk region of western Siberia.Methodology/principal findingsSamples of bile were provided by 56 study participants, half of who were infected with O. felineus, and all of who were diagnosed with gallstone disease. The microbiota of the bile was investigated using high throughput, Illumina-based sequencing targeting the prokaryotic 16S rRNA gene. About 2,797, discrete phylotypes of prokaryotes were detected. At the level of phylum, bile from participants with opisthorchiasis showed greater numbers of Synergistetes, Spirochaetes, Planctomycetes, TM7 and Verrucomicrobia. Numbers of > 20 phylotypes differed in bile of the O. felineus-infected compared to non-infected participants, including presence of species of the genera Mycoplana, Cellulosimicrobium, Microlunatus and Phycicoccus, and the Archaeans genus, Halogeometricum, and increased numbers of Selenomonas, Bacteroides, Rothia, Leptotrichia, Lactobacillus, Treponema and Klebsiella.Conclusions/significanceOverall, infection with the liver fluke O. felineus modified the biliary microbiome, increasing abundance of bacterial and archaeal phylotypes.
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