Felineus Infection Research Articles

Inhibition of Opisthorchis felineus glutathione-dependent prostaglandin synthase by resveratrol correlates with attenuation of cholangiocyte neoplasia in a hamster model of opisthorchiasis

Food-borne trematodiases represent major neglected parasitic diseases. Trematodes of the family Opisthorchiidae including Opisthorchis felineus, Opisthorchis viverrini and Clonorchis sinensis are ranked eight on the global list of the 24 most prevalent food-borne parasites. Chronic O. felineus infection symptoms include precancerous lesions with the potential for malignancy. In recent decades, liver flukes of the family Opisthorchiidae have been extensively scientifically explored, however despite this the molecular mechanisms of O. felineus pathogenicity and its carcinogenic potential have not been studied. Opisthorchis felineus glutathione-dependent prostaglandin synthase (GST σ) is the major component of the excretory–secretory product of this liver fluke. We hypothesised that the activity of this enzyme is involved in the infection pathogenesis, including the formation of precancerous lesions. To test this hypothesis and to gain insights into the mechanisms of precancerous lesion formation, we (i) investigated whether excretory parasitic GST σ retains its enzymatic activity, (ii) tested resveratrol (RSV) as a possible inhibitor of this enzyme, and (iii) assessed biliary neoplasia and oxidative DNA damage as well as the expression of neoplasia and fibrogenesis marker genes after prolonged administration of RSV in a hamster model. RSV was found to inhibit GST σ enzymatic activity in a dose-dependent manner (R = 0.85, P < 0.001; half-maximal effective dose (ED50) = 48.6 μM). Prolonged administration of RSV significantly suppressed high-grade biliary neoplasia (P = 0.008), attenuated upregulation of hyperplasia and fibrogenesis-related genes (Tgfb, α-SMA and CK7), and decreased the elevated oxidative DNA damage. Taking into account that RSV can influence a wide range of pathways, further research is needed to confirm the role of GST σ in O. felineus pathogenicity. Nevertheless, the chemopreventive effect of RSV targeting biliary neoplasia formation might be useful for improving the outcomes in infected populations and represents a compelling rationale for RSV testing in combination chemotherapy of opisthorchiasis.

Prevalence and pathology of fish trematode Opisthorchis felineus in domestic cats in Mymensingh, Bangladesh

Background: Opisthorchis felineus infection is important both for health of cats and health of people. Objective of the research was to determine the prevalence of Opisthorchis felineus and the pathological lesions caused by Opisthorchis felineus infection in domestic and stray cats in Mymensingh district, Bangladesh. Methods: A total of 36 cats were examined. The study was carried out in the Department of Parasitology and the Department of Pathology, Bangladesh Agricultural University, Mymensingh, Bangladesh. Results: Out of 36 examined cats, only 30 (83%) were infected with Opisthorchis felineus. A lower prevalence was observed in young cats &lt;1 year old (66.66%) followed by adult cats &gt;1 year (91.66%). Older cats were 5.5 times more susceptible to Opisthorchis felineus infection than that of young cats. Females (85.0%) were 1.30 times more susceptible to Opisthorchis felineus infection than that of males (81.25%). The prevalence of Opisthorchis felineus was very high all the year round. The rate of infection was 92.00%, 83.33% and 72.72% during summer, rainy and winter seasons respectively. At necropsy, the affected liver was apparently normal in size and shape. But in one case, paler and cream-colourcd area were found on the surface of liver. This liver was infected with 376 Opisthorchis felineus. Microscopically, fatty changes and mild hepatitis were detected. Conclusion: The present study indicates that Opisthorchis felineus infection in domestic and stray cats is a problem irrespective of age, sex and season of the year.

Characteristics of liver fibrosis associated with chronic Opisthorchis felineus infection in Syrian hamsters and humans

The food-borne liver trematode Opisthorchis felineus causes severe liver damage, including fibrosis. This study shows a comparison of the characteristics between cholangiofibrosis and periductal fibrosis in infected people and in the golden hamster as an experimental model. Comparative evaluation was carried out regarding collagen composition, the number of basic-producing cells, and extracellular-matrix degradation. The results revealed that characteristics of chronic opisthorchiasis due to O. felineus infection in humans and in Syrian hamsters are similar and include well-pronounced development of fibrotic complications in the liver parenchyma. Besides, a difference in fibrogenesis development was demonstrated between chronic O. felineus infection and noninfectious cholecystitis. In this study, we for the first time compared fibrogenesis between humans and model animals against the background of chronic O. felineus infection.

Opisthorchis felineus infection provokes time-dependent accumulation of oxidative hepatobiliary lesions in the injured hamster liver.

Opisthorchiasis caused by food-borne trematode Opisthorchis felineus is a substantial public health problem, with 17 million persons infected worldwide. This chronic disease is associated with hepatobiliary inflammation, cholangiocyte dysplasia, cholangiofibrosis, intraepithelial neoplasia, and even cholangiocarcinoma among chronically infected individuals. To provide first insights into the mechanism by which O. felineus infection causes precancerous liver lesions, we investigated the level of oxidative stress (lipid peroxidation byproducts and 8-hydroxy-2′-deoxyguanosine) as well as the time course profiles of chronic inflammation and fibrogenesis markers in the dynamics of opisthorchiasis from 1 month to 1.5 years postinfection in an experimental model based on golden hamsters Mesocricetus auratus. For the first time, we showed that O. felineus infection provokes time-dependent accumulation of oxidative hepatobiliary lesions in the injured liver of hamsters. In particular, over the course of infection, lipid peroxidation byproducts 4-hydroxynonenal and malondialdehyde were upregulated; these changes in general correlate with the dynamics of hepatic histopathological changes. We detected macrophages with various immunophenotypes and elevated levels of CD68, COX2, and CD163 in the O. felineus–infected animals. Meanwhile, there was direct time-dependent elevation of TNF-α (R = 0.79; p < 0.001) and CD163 protein levels (R = 0.58; p = 0.022). We also provide quantitative data about epithelial hyperplasia marker CK7 and a marker of myofibroblast activation (α smooth muscle actin). Our present data provide first insights into the histopathological mechanism by which O. felineus infection causes liver injuries. These findings support the inclusion of O. felineus in Group 1 of biological carcinogens.

METACERCARIAE OPISTHORCHIDS IN COMMERCIAL CARP FISH FROM THE LOWER AND MIDDLE OB

This article shows the infection of commercial cyprinids from the Middle and Lower Ob with muscle trematodes that are dangerous to humans, collected in 2016, 2017 and 2018. From the point of view of human health, the study of carp fish infected with opistorchids is important. It is in the Ob – Irtysh basin that the world's largest opisthorchiasis is located. In the study of carp fish for infection with muscle trematodes, a compression method was used. Comparing the infection of carp fishes of different species with metacercariae opistorchids from different landscape-geographical zones, one can speak of the leading role of ide in maintaining the Ob - Irtysh center of opisthorchiasis.The role of the bream in maintaining the West Siberian focus of opisthorchiasis is gradually increasing due to the ever-growing herd of this fish. O. felineus and M. biliside infestation is most severe in Middle Ob, in the central taiga zone. In another subzone - the northern taiga (v. Hills) and the forest-tundra zone (v. Aksarka), the extensiveness of the O. felineus infection is either equal to 100% or approaches it. The intensity of infection in the ide from Middle Ob found maximum values. In other, more northern subzones, the intensity of infection with O. felineus metacercaria was significantly inferior to the infection of the Middle Ob’s ide. Based on the above, we can talk about the greatest infection of carp fish in Middle Ob. The center of the opisthorchiasis is situated exactly here.

Effects of supramolecular complexation of praziquantel with disodium glycyrrhizinate on the liver fluke Opisthorchis felineus: An in vitro and in vivo study

Millions of people worldwide have a chronic infection with the liver fluke Opisthorchis felineus, which causes opisthorchiasis in humans and animals. The only known effective drug for this disease is praziquantel (PrzQ); however, its efficacy is below 100%, and it has some adverse effects. In this study, a water-soluble complex of PrzQ with a disodium salt of glycyrrhizic acid (disodium glycyrrhizinate; Na2GA) in a 1:10 ratio (PrzQ:GA) allowed the PrzQ dose to be decreased 11-fold for effective killing of O. felineus. An in vitro experiment showed a sufficient anthelmintic efficiency of PrzQ:GA against both adult and juvenile O. felineus individuals. A Syrian golden hamster model of opisthorchiasis revealed a considerable anthelmintic effect at all tested PrzQ:GA doses (50, 100, 200, 400, and 1100 mg/kg) with the best performance at 400 and 1100 mg/kg. Further comparison of the effects of PrzQ (400 mg/kg) and PrzQ:GA (400 mg/kg) regarding the state of the host indicated significant advantages of the latter. Histological examination showed that PrzQ:GA was better at decreasing the O. felineus–induced inflammatory infiltration (as compared with PrzQ alone) as well as interfered with the development of epithelial dysplasia and metaplasia in large bile ducts and cholangiofibrosis. Both PrzQ and PrzQ:GA decreased the number of myeloid (CFU-GM) and erythroid (BFU-E + CFU-E) colonies induced by O. felineus infection. The drugs had no negative effect on the animal behavior in an open field test 2–4 h after administration. Thus, PrzQ:GA proved to be a good anthelmintic agent having no evident adverse effects on the host, thereby suggesting that further preclinical and clinical trials would be warranted.

Imbalance in the glutathione system in Opisthorchis felineus infected liver promotes hepatic fibrosis

Although data on oxidative stress during liver fluke infection have been previously presented, a comprehensive study of the glutathione system that plays a crucial role in scavenging of reactive oxygen species (ROS) and detoxification of primary and secondary oxidation products has not been addressed yet. In the present study, the hepatic glutathione system was investigated in a hamster model of experimental opisthorchiasis infection. It was shown that chronic oxidative stress in an Opisthorchis felineus infected liver, evidenced by abundant hydroperoxide accumulation, leads to strong imbalance in the hepatic glutathione system, namely the depletion of reduced form of glutathione (GSH), lowering of the GSH/GSSG ratio, and a decrease in the glutathione peroxidase and glyoxalase 1 activity. O. felineus infection provokes hepatocellular damage that results in the progression of liver fibrosis, accompanied by an increase in collagen deposition in the hepatic tissue. Modulation of hepatic GSH levels in the O. felineus infected liver through N-acetylcysteine (NAC) or l-buthionine-S, R-sulfoxinine (BSO) treatments lead to changes in expression and activity of glutathione S-transferase and glyoxalase I as well as markedly decreases or increases collagen content in the O. felineus infected liver and the severity of liver fibrosis, respectively. Thus, the glutathione system can be considered as a target for liver protection from O. felineus-induced injury.

Fundamental Approaches to Optimize the Diagnostics and Treatment of the Opisthorchis felineus Infection in Endemic Regions: Study Methodology

Fundamental Approaches to Optimize the Diagnostics and Treatment of the Opisthorchis felineus Infection in Endemic Regions: Study Methodology

Exploratory metabolomics study of the experimental opisthorchiasis in a laboratory animal model (golden hamster, Mesocricetus auratus).

BackgroundOpisthorchiasis is a parasitic infection caused by the liver flukes of the Opisthorchiidae family. Both experimental and epidemiological data strongly support a role of these parasites in the etiology of the hepatobiliary pathologies and an increased risk of intrahepatic cholangiocarcinoma. Understanding a functional link between the infection and hepatobiliary pathologies requires a detailed description a host-parasite interaction on different levels of biological regulation including the metabolic response on the infection. The last one, however, remains practically undocumented. Here we are describing a host response on Opisthorchiidae infection using a metabolomics approach and present the first exploratory metabolomics study of an experimental model of O. felineus infection.Methodology and Principal findingsWe conducted a Nuclear Magnetic Resonance (NMR) based longitudinal metabolomics study involving a cohort of 30 animals with two degrees of infection and a control group. An exploratory analysis shows that the most noticeable trend (30% of total variance) in the data was related to the gender differences. Therefore further analysis was done of each gender group separately applying a multivariate extension of the ANOVA—ASCA (ANOVA simultaneous component analysis). We show that in the males the infection specific time trends are present in the main component (43.5% variance), while in the females it is presented only in the second component and covers 24% of the variance. We have selected and annotated 24 metabolites associated with the observed effects and provided a physiological interpretation of the findings.ConclusionsThe first exploratory metabolomics study an experimental model of O. felineus infection is presented. Our data show that at early stage of infection a response of an organism unfolds in a gender specific manner. Also main physiological mechanisms affected appear rather nonspecific (a status of the metabolic stress) the data provides a set of the hypothesis for a search of the more specific metabolic markers of the Opisthorchiidae infection.

Life by the river: neglected worm infection in Western Siberia and pitfalls of a one-size-fits-all control approach

The One Health movement aims to provide integrated responses to problems that emerge at the intersections of human, animal, and ecological health. However, it risks derailment due to over-reliance on top-down global responses and generalisations that often fail to fit real-world settings. In this article, we examine the case of parasitic Opisthorchis felineus infection in Western Siberia and argue that understanding the local social dynamics of disease exposure and transmission, and how people live their lives in interactions with other species is crucial for making One Health work in practice. This qualitative study was conducted in preparation for developing an opisthorchiasis control programme and involved 20 in-depth interviews, participant observation, and multiple unstructured interviews in the village of Melnikovo. We explored the social dynamics of O. felineus transmission and designed a participatory model of these dynamics. This model attests to the specificity of transmission dynamics in Western Siberia and demonstrates the importance of understanding this public health issue as embedded in social networks and animated by a variety of locally-specific linkages between ways of life, food and health cultures, health-care systems, economies, and landscapes. Our work highlights that such participatory approaches have to be an integral part of One Health interventions if these interventions are to be effective and legitimate.

Magnetic resonance imaging and spectroscopy for differential assessment of liver abnormalities induced by Opisthorchis felineus in an animal model.

BackgroundEuropean liver fluke Opisthorchis felineus, causing opisthorchiasis disease, is widespread in Russia, Ukraine, Kazakhstan and sporadically detected in the EU countries. O. felineus infection leads to hepatobiliary pathological changes, cholangitis, fibrosis and, in severe cases, malignant transformation of bile ducts. Due to absence of specific symptoms, the infection is frequently neglected for a long period. The association of opisthorchiasis with almost incurable bile duct cancer and rising international migration of people that increases the risk of the parasitic etiology of liver fibrosis in non-endemic regions determine high demand for development of approaches to opisthorchiasis detection.Methodology/Principal findingsIn vivo magnetic resonance imaging and spectroscopy (MRI and MRS) were applied for differential assessment of hepatic abnormalities induced by O. felineus in an experimental animal model. Correlations of the MR-findings with the histological data as well as the data of the biochemical analysis of liver tissue were found. MRI provides valuable information about the severity of liver impairments induced by opisthorchiasis. An MR image of O. felineus infected liver has a characteristic pattern that differs from that of closely related liver fluke infections. 1H and 31P MRS in combination with biochemical analysis data showed that O. felineus infection disturbed hepatic metabolism of the host, which was accompanied by cholesterol accumulation in the liver.ConclusionsA non-invasive approach based on the magnetic resonance technique is very advantageous and may be successfully used not only for diagnosing and evaluating liver damage induced by O. felineus, but also for investigating metabolic changes arising in the infected organ. Since damages induced by the liver fluke take place in different liver lobes, MRI has the potential to overcome liver biopsy sampling variability that limits predictive validity of biopsy analysis for staging liver fluke-induced fibrosis.

Infection with Opisthorchis felineus induces intraepithelial neoplasia of the biliary tract in a rodent model.

The liver fluke Opisthorchis felineus is a member of the triad of epidemiologically relevant species of the trematode family Opisthorchiidae, and the causative agent of opisthorchiasis felinea over an extensive range that spans regions of Eurasia. The International Agency for Research on Cancer classifies the infection with the liver flukes Opisthorchis viverrini and Clonorchis sinensis as group 1 agents and a major risk factor for cholangiocarcinoma. However, the carcinogenic potential of the infection with O. felineus is less clear. Here, we present findings that support the inclusion of O. felineus in the Group 1 list of biological carcinogens. Two discrete lines of evidence support the notion that infection with this liver fluke is carcinogenic. First, novel oxysterol-like metabolites detected by liquid chromatography-mass spectroscopy in the egg and adult developmental stages of O. felineus, and in bile, sera, and urine of liver fluke-infected hamsters exhibited marked similarity to oxysterol-like molecules known from O. viverrini. Numerous oxysterols and related DNA-adducts detected in the liver fluke eggs and in bile from infected hamsters suggested that infection-associated oxysterols induced chromosomal lesions in host cells. Second, histological analysis of liver sections from hamsters infected with O. felineus confirmed portal area enlargement, inflammation with severe periductal fibrosis and changes in the epithelium of the biliary tract characterized as biliary intraepithelial neoplasia, BilIN. The consonance of these biochemical and histopathological changes revealed that O. felineus infection in this rodent model induced precancerous lesions conducive to malignancy.

Mechanisms of trematodiases pathogenicity: the presence of the secretory proteins from the liver fluke Opisthorchis felineus in the gallbladder tissues of the patients with chronic opisthorchiasis

Opisthorchiasis caused by the liver fluke Opisthorchis felineus infection remains a serious public health problem in the former USSR and Eastern European countries. O. felineus infests the bile ducts, the liver and gallbladder of many fish-eating mammalian species, including humans. Opisthorchiasis leads to a number of related diseases of the liver and pancreas: liver fibrosis, cholangitis, cholecystitis, liver cysts and pancreatitis. Excretory-secretory products of the parasite are considered to be key factors in host-parasite relationships and mediate pathogenic pleiotropic effects on the host organism. The aim of this study was to determine the helminthic proteins (thioredoxin peroxidase and glutathione-S-transferase) in the gallbladder tissues of the experimental animals and patients with opisthorchiasis disease. We demonstrated by immunohistochemistry assay using antibodies against recombinant O. felineus proteins that thioredoxin peroxidase and glutathione-S-transferase could be detected in the biliary duct epithelium of the experimental animals and in human gallbladder tissues. Moreover, these proteins could also be detected in human gallbladder infiltrated cells and underlying connective tissues. The results are important for understanding the molecular mechanisms of opisthorchiasis pathogenesis, as well as for improvement of the immunodiagnostics of the opisthorchiasis and opisthorchiasis-related diseases.

Opisthorchiasis in Western Siberia: Epidemiology and distribution in human, fish, snail, and animal populations

Opisthorchiasis is a widespread helminth infection in Russia. The largest opisthorchiasis endemic focus in the world is the Ob river watershed in Western Siberia. The main causative agent of this condition is the liver fluke, Opisthorchis felineus. In addition, another liver fluke species in the Opisthorchiidae family, Metorchis bilis, causes a symptomatically similar disease, metorchiasis. Despite a long research history going back to 1927, opisthorchiasis remains a serious problem in Russia, and numerous questions related to the epidemiology of these liver fluke infections and their patterns of distribution in Western Siberia, the causes of high prevalence in different populations, and the prognosis of the epidemiological situation remain to be answered.In this review, we first briefly describe the life cycle of O. felineus and then summarize the available published data on the epidemiological aspects of O. felineus infection among populations in Western Siberia. Additionally, the geographical distribution and rates of infection with the two major small liver flukes, O. felineus and M. bilis, in the intermediate (Bithyniidae snails and cyprinid fish) and definitive (humans, wild and domestic carnivorous animals and birds) hosts are described to assess their role in the transmission cycle. Moreover, species in the genus Opisthorchis and the genus Metorchis that have been reported in carnivorous mammals and birds in Western Siberia are listed and their potential to serve as the agents of opisthorchiasis transmission is discussed.

Opisthorchis felineus infection and cholangiocarcinoma in the Russian Federation: A review of medical statistics

Opisthorchis felineus (O. felineus) occurs in Western Siberia and many other parts of the Russian Federation (RF). The true extent of its distribution is not known. Chronic infection may lead to severe hepatobiliary morbidity. According to surgical and experimental reports, long-term infestation might significantly increase the risk for cholangiocarcinoma (CCA). To date, no association between O. felineus infection and CCA has been demonstrated. The objective of this study was to review existing health data on the incidence of O. felineus infection and on the incidence of CCA in the RF. We reviewed the official medical statistics on reported O. felineus infection and CCA in 83 political/geographical units of the RF, covering the period January 2011–December 2013. Annual incidence data were obtained from Rospotrebnadzor and from official medical statistics. We calculated the average annual incidence of infection and cancer. The average annual incidence of O. felineus was 24.7±9.0 cases per 100,000 population. The highest incidence was observed in Khanty-Mansiysk district (599.7 cases per 100,000 population per year). In 27 geographical units, no O. felineus cases were reported. The incidence of liver and intrahepatic bile duct cancers was 4.8±0.2 cases per 100,000 population; the highest rate was reported in Sakha Republic and Tomsk Oblast (14.5 and 9.3 cases per 100,000 population), and the lowest in Yamalo-Nenets Autonomous Okrug (0.9 cases per 100,000 population). O. felineus incidence was not associated with the mean annual incidence of liver and intrahepatic bile duct cancers (r=0.20, p=0.07). This study documents the importance of opisthorchiasis in certain endemic areas and presents the best available data on associations between O. felineus infection and liver/intrahepatic bile duct cancers in RF. The findings support the need to implement a public health control programme against liver fluke infections and to increase the availability of anthelmintic treatment. Further studies are warranted to assess the contribution of opisthorchiasis to the CCA in RF.

Experimental opisthorchiasis: a study of blood cells, hematopoiesis and startle reflex in laboratory animals

One of the species of the family Opisthorchiidae, Opisthorchis felineus (O. felineus), causes severe disturbances in humans and animals, and so it is the subject of important research studies. Two weeks after infection we compared the impact of O. felineus invasion on the changes in blood cells composition, bone marrow hematopoiesis and behavioral startlereflex in inbred C57BL/6 male mice and Syrian hamsters (Mesocricetus auratus). Considerable interspecies differences were revealed for many parameters estimated. It was found that the relative weight of the main organ of the peripheral immune system – spleen, is significantly larger in mice than in hamsters. Moreover, the infection with O. felineus caused a significant enlargement of the spleen only in mice. More pronounced changes in the blood cells composition, which was accompanied by activation of hematopoietic stem cells of myeloid and erythroid set, were determined in hamsters. Blood changes in the response to infection in mice were less severe and were not accompanied by the changes in colony formation. Mouse acoustic startle reaction differed from hamster one too. The expression of the startle reaction and the value of pre-pulse inhibition were discriminated in animals of two species. Infected hamsters had no reaction of habituation to the sound stimulus. In addition, the maturation of O. felineus worms was faster in hamsters than in mice. Data obtained suggest a greater resistance of mice to O. felineus infection, but do not exclude the availability of mice as a model in the study of processes taking place in the host during the development of experimental opisthorchiasis.

Analysis of Opisthorchis felineus infection and liver and intrahepatic bile ducts cancer incidence rate in Russian Federation

The purpose of this paper is to perform the pilot review of the official medical statistics on reported Opisthorchis felineus (O. felineus) infection and liver and intrahepatic bile duct cancer in Russian Federation, covering the period 2011–2013. Materials and methods. Statistical data collected from 81 regions of Russia, reflecting the incidence of opisthorchiasis over a period from the 1st of January 2011 to the 31st of December 2013 was analyzed. The analysis of official health statistics of malignant hepatic lesions and intrahepatic bile duct was conducted in 82 regions of Russia over the same period. The data was obtained from the annual reports of the Russian center of information technologies and epidemiological studies in the field of oncology. Results. The average annual incidence of O. felineus was 24,7 ± 9,0 cases per 100,000 population. The highest incidence was observed in Khanty-Mansiysk Autonomous Okrug, Yamalo-Nenets Autonomous Okrug, Tyumen Oblast, Tomsk Oblast, Omsk Oblast and Novosibirsk Oblast. We did not observe any significant change in O. felineus infection rates within last 20 years. However, we found that infection incidence had increased in some geographical units of Western Siberia. The incidence of liver and intrahepatic bile duct cancers was 4,8 ± 0,2 cases per 100,000 population; the highest rate was reported in Far East regions, Siberia and Volgograd Oblast. This study documents the importance of opisthorchiasis in certain endemic areas and presents the best available data on associations between O. felineus infection and liver/intrahepatic bile duct cancers in endemic areas of Russian Federation.

P158: Metabolic profiling of bile juice from the patients with cholangiocarcinoma-associated diseases

Background Opisthorchiasis is a form of foodborne trematodiasis which is caused by liver flukes. It has been shown that a chronic Opisthorchiasis infection increases a risk of cholagiocarcinoma of liver. It is commonly believed that a gradual change of homeostasis in a parasite microenvironment (bile) leads to liver fluke-induced cancer. Nevertheless, no systematic, analytically driven studies confirming this hypothesis have been published yet. The restricted access to clinical material and extreme complexity of the biological matrix (bile) both are the important “rate limiting factors” for a progress in the field. Here we present for the first time a cross-platform mass spectrometric analysis of bile juice collected from the patients with cholangiocarcinoma-associated diseases. We show that an effective analysis of such complex biological matrix as bile juice requires a combination of orthogonal analytical platforms (e.g. RPLC–MS and HILIC–MS) maximizing coverage of the metabolic space. Materials and methods 28 patients with O. felineus infection and 30 negative controls were included in the study. The infection status was confirmed using microscopy analysis of the bile. Bile samples were collected from the gallbladder using sterile puncture, directly frozen and stored at −80 °C until analysis. The samples were randomized and organized into the acquisition blocks consisting of the samples and quality controls (QC). Experiments were carried out with a Dionex Ultimate 3000 LC system (Thermo Scientific/Dionex, The Netherlands) equipped with a Dual Gradient Separation pump allowing for parallel LC analysis, and hyphenated to an Impact UHR-qTOF mass analyzer (Bruker Daltonics, Germany). Reversed-phase experiments (RPLC) were performed with an UHPLC BEH Shield RP18 column 100 × 2.1 mm, 1.7 μm (Waters) and HILIC experiments with a Luna HILIC column (Phenomenex, The Netherlands) of 100 × 2.00 mm, 3 μ m. RPLC data were acquired in ESI positive mode and HILIC in negative mode, respectively. The data acquisition rate was set to 1 Hz over a mass range of m / z 50–1000. The LC–MS data files were aligned by using the in-house developed alignment algorithm MS-Align 2 tool ( www.ms-utils.org/msalign2 ). Results After the data prepressing, which includes alignment, noise filtering and peak picking two data matrixes costing of 412 features (metabolites) for RPLC and 428 ones for HILIC were generated. To evaluate a degree of similarity between the two data matrixes the RV coefficient (a multivariate extension of correlation coefficient) was used. The coefficient has flattened at 0.58 showing that despite a strong overlap between the datasets there is a substantial number of the “platform specific” metabolites. Those structures will certainly be missed if a single platform strategy is applied. Conclusion Here we present for the first time a cross-platform mass spectrometric analysis of bile juice collected from the patients cholangiocarcinoma-associated diseases. We show that a combination of the two platforms greatly improves the coverage of the metabolome and as such should be a firstchoice for exploratory studies of the complex biological matrixes. Work was conducted with the application of the Tomsk regional common use center technical equipment acquired thanks to a grant of the Russian Ministry of the Agreement No. 14.594.21.0001 (RFMEFI59414X0001). This project is supported by “The Tomsk State University Academic D.I. Mendeleev Fund Program” under Grant (No. 18.1.52.2015).

Effect of Opisthorchis felineus infection and dimethylnitrosamine administration on the induction of cholangiocarcinoma in Syrian hamsters

The food-borne liver trematode Opisthorchis felineus is an emerging source of biliary tract diseases on the territory of the former Soviet Union and Eastern Europe. This parasite along with trematodes Opisthorchis viverrini and Clonorchis sinensis belong to the triad of epidemiologically important liver flukes of the Opisthorchiidae family.It is known that O. viverrini and С. sinensis are the main risk factors of cholangiocarcinoma (CCA) in the endemic regions. The carcinogenic potential of О. felineus has not been well researched because of the absence of systematic pathomorphological, clinical, and epidemiological studies on O. felineus opisthorchiasis.In the present study, we show the results of detailed histopathological analysis and comprehensive evaluation of inflammation, bile duct dysplasia, periductal fibrosis, bile duct hyperplasia, bile duct proliferation, egg granuloma, cysts, cholangiofibrosis, and CCA from 10 to 30weeks following infection of Syrian hamsters with O. felineus accompanied by oral administration of dimethylnitrosamine (DMN). The results revealed that O. felineus contributes to bile duct cancer development in the hamster model. During the combined action of O. felineus and DMN, morphological features of the liver underwent dramatic changes at the cellular and organ levels. Already in the early stages of the experiment, we observed extensive periductal fibrosis, active inflammation, proliferation of the bile duct, bile duct dysplasia and egg granulomas. Later, against the background of all these changes, cholangiofibrosis and CCA were found.Our work is the first step in the study of carcinogenic potential of O. felineus. Obtained data indicate the risk of CCA of patients having chronic О. felineus opisthorchiasis, and underscore the need for the development of programs for control of this helminthiasis.

&lt;i&gt;Opisthorchis felineus&lt;/i&gt; Infection and Harmful Metals Bio-Concentration: A Pilot Study in Hamster Model

Nowadays, the question about the negative impacts of liver fluke on the host organism and mechanisms of this damage is open. The bio-concentration of some heavy and toxic metals in tissue of adult forms of Opisthorchis felineus and its accumulation in liver tissue and cardiac muscle in the Syrian Golden Hamster (Mesocricetus auratus) model was analyzed. It is outlined, that Opisthorchis felineus infection leads to accumulating aluminum in a liver tissue of the host. It was also found, that adult fluke bio-concentrates iron and aluminum in their tissue. The shortage of such essential elements as Mn, Zn and Сu in infected organism is discussed.