Alveolar echinococcosis is a worldwide zoonosis of great public health concern. Analysis of genome data for Echinococcus multilocularis has identified antigen families that can be used in diagnostic assays and vaccine development. However, little gene expression data is available for antigens of the egg and early larval stages. To address this information gap, we used a Next-Generation Sequencing approach to investigate three different stages (non-activated and activated oncospheres, and early stage metacestodes) of E. multilocularis (Nemuro strain). Transcriptome data analysis revealed that some diagnostic antigen gp50 isoforms and the antigen Eg95 family dominated in activated oncospheres, and the antigen B family dominated in early stage metacestodes. Furthermore, heat shock proteins and antigen II/3 are constantly expressed in the three stages. The expression pattern of various known antigens in E. multilocularis may give fundamental information for choosing candidate genes used in diagnosis and vaccine development.
Cl 2 -ATPase in the CNS is a candidate for an outwardly directed neuronal Cl 2 transporter requiring phosphatidylinositol-4-phosphate (PI4P) for its optimal activity. To test its pathophysiological changes in a phosphatidylinositol (PI) metabolism disorder, the effects of neurotoxic factors in Alzheimer's disease (AD), amyloid b proteins (Abs), on the Cl 2 -ATPase activity were examined using primary cultured rat hippocampal neurons. Amyloid b proteins (1±40, 1±42 and 25±35) concentration-dependently (1±100 nM) and timedependently (from 1 h to 6 day) decreased Cl 2 -ATPase activity and elevated intracellular Cl 2 concentrations ([Cl 2 ] i ), Ab25±35 being the most potent. Addition of inositol or 8-Brcyclic GMP completely reversed these Ab-induced changes. The recoveries in enzyme activity were attenuated by an inhibitor of PI 4-kinase, 10 mM wortmannin or 20 mM quercetin, but not by a PI 3-kinase inhibitor, 50 nM wortmannin or 10 mM LY294002. The PI, PIP and PIP2 levels of the plasma membrane-rich fraction were lower in the Ab-treated cells as compared with each control. In the Ab-exposed culture, but not in control, stimulation by 10 mM glutamate for 10 min signi®cantly increased fragmentation of DNA and decreased cell viability. Addition of inositol or 8-Br-cyclic GMP prevented the effect of Ab-treatment on the neurotoxicity of glutamate. Thus, Abs reduce neuronal Cl 2 -ATPase activity, resulting in an increase in [Cl 2 ] i probably by lowering PI4P levels, and this may re¯ect a pre-apoptotic condition in early pathophysiological pro®les of AD.
The cotton rat (Sigmodon hispidus) is a laboratory rodent that has been used for studies on human infectious diseases. In the present study, we observed that female cotton rats, not the male cotton rats, developed chronic anemia characterized by reduced red blood cell, hemoglobin, and hematocrit levels from 5 to 9 months of age without any changes in the mean corpuscular hemoglobin and volume levels. In peripheral blood, the reticulocyte count did not increase in response to anemia in female cotton rats, and no extramedullary hematopoiesis was observed in the liver or spleen. Further, the serum levels of urea nitrogen and creatinine increased from 5 to 9 months of age in female cotton rats compared to male cotton rats, and these increases became more prominent from 10 months of age onward, indicating chronic kidney disease. Histopathologically, female cotton rats manifested tubulointerstitial lesions characterized by the infiltration of mononuclear cells, including plasma cells and CD3(+) T-cells, as well as the dilation of calbindin-D28k(+) distal tubules from 5 to 9 months of age. The severity of these lesions progressed from 10 months of age onward, and renal fibrotic features and numerous tubular cysts appeared without any obvious glomerular lesions. A significant decrease in the erythropoietin protein levels was observed in the kidney of aged female cotton rats, and significant correlations were detected between anemia and tubulointerstitial damage. These results suggest that aged female cotton rats chronically develop renal anemia, and this rodent may serve as a novel model to elucidate its pathogenesis.
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