The formation of host-parasite relationships in an experimental mixed pathology of opisthorchiasis-tuberculosis as dependent on the phase of the Opisthorchis infestation]

Superinfection of animals having Opisthorchis invasion with Mycobacterium tuberculosis leads to destabilization of host-parasite relationships in opisthorchiasis and formation of new host-parasitocenotic interrelations whose manifestation depends on the phase of the invasive process. At the acute invasive phase of mixed pathology (2 weeks) the activity of the host's immune system increases, while the biological activity of helminths and the number of M. tuberculosis colonies decrease. And on the contrary, at the subacute phase of mixed pathology (2.5 months) the activity of the host's immune system decreases, while the reproductive activity of helminths and the isolation rate of M. tuberculosis from pulmonary tissue increases in comparison with the groups of animals with monoinvasion and monoinfection.     

Susceptibility of Bithynia inflata mollusks from different populations to Opisthorchis felineus infestation

The paper reports the results of experimental infection of a specific intermediate host, molluscs Bithynia inflata from differently susceptible populations, with Opisthorchis felineus in one opisthorchis nidus. Non-susceptibility of molluscs from resistant and hyperinfected populations to O. felineus infection is shown.     

Comparative histopathology of Opisthorchis felineus and Opisthorchis viverrini in a hamster model: an implication of high pathogenicity of the European liver fluke

European liver fluke (Opisthorchis felineus) and Asian liver fluke (Opisthorchis viverrini) are similar in morphology but comparative pathology of the infections has not been described. We therefore did comparative histopathology of both parasites in an experimental animal model. The study was conducted in 3 groups of 105 Syrian golden hamsters; the first and second groups fed with 50 metacercariae of O. felineus (OF) or O. viverrini (OV) and the last group was uninfected controls. Five hamsters in each group were euthanized on weeks 1, 2, 4, 8, 12 and 24 post-infection. The liver tissue was fixed and processed for routine histopathology and immunohistochemistry for proliferation markers (BrdU or PCNA). Overall, the liver histopathology of O. felineus and O. viverrini infection was generally similar. However, various histopathogical features including intense inflammation, fibrosis, biliary and goblet cell hyperplasia and dysplasia occurred earlier in the OF group. In addition, the existence of precancerous lesions such as cholangiofibrosis in a long-term infection was observed only in this group. O. felineus is larger in size than O. viverrini which, together with its excreted and secreted antigens, likely is crucial in the induction of liver fluke induced disease. The differences in nature and timing of the histopathological profile indicate that opisthorchiasis caused by the European liver fluke O. felineus is more pathogenic than its Asian relative O. viverrini.     

Infection with the carcinogenic human liver fluke, Opisthorchis viverrini

Throughout Southeast Asia there is a strikingly high incidence of cholangiocarcinoma (CCA--hepatic cancer of the bile duct epithelium), particularly in people from rural settings in Laos and Northeast Thailand who are infected with the liver fluke, Opisthorchis viverrini, one of only three carcinogenic eukaryotic pathogens. More ubiquitous carcinogenic microbes, such as Helicobacter pylori, induce cancer in less than 1% of infected people, while as many as one-sixth of people with opisthorchiasis will develop CCA. The mechanisms by which O. viverrini causes cancer are multi-factorial, involving mechanical irritation from the activities and movements of the flukes, immunopathology, dietary nitrosamines and the secretion of parasite proteins that promote a tumourigenic environment. Genomic and proteomic studies of the liver fluke secretome have accelerated the discovery of parasite proteins with known/potential roles in pathogenesis and tumourigenesis, establishing a framework towards understanding, and ultimately preventing, the morbidity and mortality attributed to this highly carcinogenic parasite.     

8-nitroguanine formation in the liver of hamsters infected with Opisthorchis viverrini

Nucleic acid damage by reactive nitrogen and oxygen species may contribute to the carcinogenesis associated with chronic infection and inflammation. We examined 8-nitroguanine and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) formation and nitric oxide (NO) production in hamsters infected with Opisthorchis viverrini (OV). Formation of 8-nitroguanine was assessed immunohistochemically with an antibody specific for 8-nitroguanine. 8-nitroguanine formation was found mainly in the cytoplasm and slightly in the nucleus of inflammatory cells and epithelial lining of bile duct at inflammatory areas in the liver. 8-nitroguanine immunoreactivity reached the highest intensity on day 30. A time profile of 8-nitroguanine formation was closely associated with that of plasma nitrate/nitrite. HPLC with an electrochemical detector revealed that the amount of 8-oxodG in the liver reached the maximal level on day 21. The mechanisms of 8-oxodG and 8-nitroguanine formation via O2*- and NO production triggered by OV infection were discussed in relation to cholangiocarcinoma development.     

Susceptibility of Bithynia inflata mollusks from discrete populations for Opisthorchis felineus infestation from different foci of opisthorchiasis

Experiments were conducted on crossed infection of the specific intermediate host of Opisthorchis, molluscs of Bithynia inflata from different and remote populations, with eggs of O. felineus from different nidi of the disease. The strains of the agent were found to be heterogeneous that is expressed in different degree of compatibility with the intermediate host.     

Effect of growth hormone on rat liver N-acetyl-L-glutamate

Earlier studies have revealed, upon hypophysectomy, a specific increase in mitochondrial urea cycle enzymes, namely carbamyl phosphate synthetase and ornithine transcarbamylase. Administration of growth hormone to hypophysectomized rats brought these enzyme activities back to normal. Since growth hormone plays a role in the formation of citrulline and ultimately urea, in the present study its effect on the levels of N-acetyl-L-glutamate, an allosteric activator of carbamyl phosphate synthetase has been investigated. A significant increase in N-acetyl-L-glutamate concentration in rat liver on hypophysectomy and its reversal back to normal levels on growth hormone administration was reported. These results suggest that the lack of growth hormone tends to amplify urea production by the liver.     

Effects of gall midge infestation on plant growth and growth regulating substances

The biological association between plant and gall-inducing insects can be characterized by growth and developmental abnormalities of the plant tissues caused by the feeding activities of the insects. The relationship between Wachtliella persicariae L. (Diptera: Cecidomyiidae) and Polygonum amphibium L. has been studied.The gall-making larvae settle only on the undifferentiated parts of the leaf and prevent the normal unfolding of the lamina; and there is a close developmental synchronization between the two organisms.The essential reaction of the leaf tissue to the continuous feeding activity of the larvae is primarily a disturbance of the normal development, characterized by an imbalance in growth-regulating substances. Bioassays showed greater amounts of auxin-like substances in the galled tissues. The way in which feeding behaviour, feeding activity and nutritional physiology possibly interfere with the growth-regulating factors of the leaf is discussed.The resulting abnormal leaf growth and development is essential for the normal growth and development of the larvae.

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Zusammenfassung Die biologische Verbindung zwischen Pflanzen und gallenerzeugenden Insekten kann durch Wachstums-und Entwicklungsabnormalitäten der Pflanzengewebe gekennzeichnet werden, welche durch die Fraßtätigkeit der Insekten hervorgerufen werden. Hier wurde die Beziehung zwischen Wachtliella persicariae (Diptera: Cecidomyiidae) und Polygonum amphibium untersucht.Die gallenerzeugende Larve siedelt sich nur auf undifferenzierten Teilen des Blattes an und verhindert die normale Entfaltung der Lamina; es besteht eine enge Synchronisation der Entwicklung beider Organismen.Die wesentliche Reaktion des Blattgewebes auf die fortdauernde Fraßaktivität der Larve stellt zunächst eine Störung der normalen Entwicklung dar, die durch ein Ungleichgewicht der Stoffe charakterisiert ist, die das Pflanzenwachstum kontrollieren. Biotests ergaben in den vergallten Geweven größere Beträge auxinartiger Stoffe. Der Weg, auf dem Fraßverhalten, Fraßaktivität und Ernährungsphysiologie mit den wachstumkontrollierenden Faktoren des Blattes ineinandergreifen, wird diskutiert.Das entstehende anomale Blattwachstum und seine Entwicklung sind für das normale Wachstum und die Ontogenese der Larve wesentlich.

     

RNA interference targeting cathepsin B of the carcinogenic liver fluke, Opisthorchis viverrini

Functional genomics have not been reported for Opisthorchis viverrini or the related fish-borne fluke, Clonorchis sinensis. Here we describe the introduction by square wave electroporation of Cy3-labeled small RNA into adult O. viverrini worms. Adult flukes were subjected to square wave electroporation employing a single pulse for 20 ms of 125 V in the presence of 50 μg/ml of Cy3-siRNA. The parasites tolerated this manipulation and, at 24 and 48 h after electroporation, fluorescence from the Cy3-siRNA was evident throughout the parenchyma of the worms, with strong fluorescence evident in the guts and reproductive organs of the adult worms. Second, other worms were treated using the same electroporation settings with double stranded RNA targeting an endogenous papain-like cysteine protease, cathepsin B. This manipulation resulted in a significant reduction in specific mRNA levels encoding cathepsin B, and a significant reduction in cathepsin B activity against the diagnostic peptide, Z-Arg-Arg-AMC. This appears to be the first report of introduction of reporter genes into O. viverrini and the first report of experimental RNA interference (RNAi) in this fluke. The findings indicated the presence of an intact RNAi pathway in these parasites which, in turn, provides an opportunity to probe gene functions in this neglected tropical disease pathogen.     

Effect of epidermal chalones on induction and growth of tumors of the cervicovaginal epithelium of mice

Tumors of the cervicovaginal epithelium were induced in BALB/c female mice with local application of dimethylbenzanthra cene. In the course of inducing tumors or after their appearance the mice were treated intravaginally with 55-81% ethanolic extract of the rat skin or liver (control) containing 1% chalones. The skin extract rather than liver one delays slightly the growth and incidence of cervical and vaginal tumours, and increases the survival of animals.     

Proliferation of the intestinal epithelium and of the regenerating liver with a deficiency of the epidermal growth factor

To determine the role of the epidermal growth factor (EGF) in the regenerative stimulation of intestinal epithelium and hepatocyte proliferation after partial resection of these organs the labeling index of the intestine and liver in sialadenectomized rats was studied. EGF concentration in the saliva and serum was determined using radioimmunoassay. The decrease in EGF concentration after the removal of submandibular salivary glands was shown to slower hepatocytes entering the mitotic cycle and to inhibit the activity of enterocyte proliferation in the small intestine. The data show pronounced in vivo mitogenic effect of EGF on the liver and intestinal epithelium.