The in vivo angiogenic potential of transplanted human umbilical cord blood (UCB) CD133(+) stem cells in experimental chronic hepatic fibrosis induced by murine schistosomiasis was studied. Enriched cord blood-derived CD133(+) cells were cultured in primary medium for 3 weeks. Twenty-two weeks post-Schistosomiasis infection in mice, after reaching the chronic hepatic fibrotic stage, transplantation of stem cells was performed and mice were sacrificed 3 weeks later. Histopathology and electron microscopy showed an increase in newly formed blood vessels and a decrease in the fibrosis known for this stage of the disease. By immunohistochemical analysis the newly formed blood vessels showed positive expression of the human-specific angiogenic markers CD31, CD34 and von Willebrand factor. Few hepatocyte-like polygonal cells showed positive expression of human vascular endothelial growth factor and inducible nitric oxide synthase. The transplanted CD133(+) human stem cells primarily enhanced hepatic angiogenesis and neovascularization and contributed to repair in a paracrine manner by creating a permissive environment that enabled proliferation and survival of damaged cells rather than by direct differentiation to hepatocytes. A dual advantage of CD133(+) cell therapy in hepatic disease is suggested based on its capability of hematopoietic and endothelial differentiation.
A monoclonal antibody (mAb), 2F/11F, raised against Schistosoma haematobium soluble egg antigen (SEA) was found to be nonreactive with S. mansoni SEA or other parasite antigens (Fasciola hepatica, Echinococcus granulosus). This IgG1 mAb recognized a repetitive epitope on S. haematobium SEA in the molecular-weight regions of 70, 42, and 35 kDa. It was employed as both an antigen-capture and a biotinylated detection antibody in a sandwich enzyme-linked immunosorbent assay (ELISA) for the detection of circulating schistosome antigen (CSA) and had a detection limit of <1 ng S. haematobium SEA/ml. CSA levels were measured in serum and urine samples from 116 S. haematobium-infected rural students before therapy and at 4, 8, and 12 weeks after praziquantel treatment. Serum and urine samples from 50 S. mansoni -infected patients, 15 patients harboring other parasites, and 30 noninfected individuals were also assessed. CSA was detected in 90.5% of serum samples and 94% of urine samples from S. haematobium-infected patients. CSA was undetectable in serum from the 15 patients harboring other parasites and in 94% of serum samples and 84% of urine samples from S. mansoni-infected patients. In the S. haematobium-infected group a positive correlation was detected between CSA levels in serum and urine samples and the egg load per 10 ml urine. A significant reduction in CSA levels was detected in serum and urine samples after praziquantel therapy. CSA was undetectable in 87% of serum samples and 81.5% of urine samples from schistosomiasis haematobium patients at 12 weeks post-treatment. These data demonstrate that the use of mAb 2F/11F for detection of CSA provides a sensitive method for the immunodiagnosis and monitoring of cure of schistosomiasis haematobium.
The aim of this study is to assess the effect of the crude methanol extracts of five herbs (Pelargonium zonale, Terminalia bellerica, Philodendron selloum, Ulmus pumila and Ulmus parvifolia) on human hepatoma and murine myeloma cell lines. Assessment included in vitro neutral red cytotoxicity assay and cytopathological diagnosis.IC50 values obtained in both cell lines using neutral red assay showed that the plant extracts of both Terminalia bellerica and Philodendron selloum, were the most effective in inducing cytotoxicity in HepG2, IC50 (16.25±0.20 μg/ ml, 17.51±0.70 μg/ml respectively). The result showed that there was no significant difference in the IC50 between the different plants extract in Myeloma cell line. However, in hepatoma cell line, the IC50 of both Terminalia bellerica and Philodendron selloum were significantly lower (p< 0.01) than the IC50 of Ulmus pumila, Ulmus parvifolia and Pelorgonium zonale. The results indicated that the in vitro NR assay of cytotoxic activities induced by plant methanol extracts were supported by the cytopathological changes on the same population of cells. Conclusions:Neutral red cytotoxicity assay is a suitable test for screening anti-cancer potential of natural products materials. The present results showed that Terminalia bellerica and Philodendron selloum methanol extracts induced cytotoxicity on HepG2 cells. The identification of the effect of individual constituents of each plant methanol extract recommended. In vivo study on experimental level is needed to verify the mechanism of action.
The present study was designed to prepare monoclonal antibodies (MAbs) against Schistosoma haematobium soluble egg antigen (SEA) with immunodiagnostic potential for urinary schistosomiasis. From a panel of MAbs, a pair of IgG1 MAbs (2D/11C and 10B/2C) specific for S. haematobium SEA was selected. Both MAbs recognized one band with a 42-kDa molecular weight by western blots. The pair of MAbs was employed in sandwich ELISA for the detection of circulating schistosome antigen (CSA), one as antigen-capturing antibody and the other as peroxidase-conjugated antigen-detecting antibody. The lower detection limit of the assay was 1 ng/ml of S. haematobium SEA. The assay was performed on sera of 65 S. haematobium-infected patients, 25 patients infected with other parasites (Fasciola hepatica, Echinococcus granulosus), and 20 noninfected individuals. CSA was demonstrated in 89% of the S. haematobium-infected group. However, CSA was negative in the sera of healthy individuals and patients infected with other parasites, giving an overall specificity of 100% for the CSA assay. A positive correlation (r=0.37, p<0.01) was detected between the number of S. haematobium eggs excreted in 10 ml urine and the CSA level detected in the sera of S. haematobium-infected patients. Our data show that the use of anti-S. haematobium MAbs for the detection of CSA provides a sensitive and specific method for the immunodiagnosis of active S. haematobium-infected patients. Moreover, CSA assay using this anti-S. haematobium MAb/ELISA system was proven to correlate with intensity of infection and hence morbidity assessment.
Objectives: Liver transplant is the cornerstone line of treatment for chronic liver diseases; however, the long list of complications and obstacles stand against this operation. Searching for new modalities for treatment of chronic liver illness is a must. In the present research, we aimed to compare the effects of transplant of undifferentiated human mesenchymal stem cells, in vitro differentiated mesenchymal stem cells, and adult hepatocytes in an experimental model of chronic liver failure. Materials and Methods: Undifferentiated human cord blood mesenchymal stem cells were isolated, propagated, and characterized by morphology, gene expression analysis, and flow cytometry of surface markers and in vitro differentiated into hepatocytelike cells. Rat hepatocytes were isolated by double perfusion technique. An animal model of chronic liver failure was developed, and undifferentiated human cord blood mesenchymal stem cells, in vitro hepatogenically differentiated mesenchymal stem cells, or freshly isolated rat hepatocytes were transplanted into a CCL4 cirrhotic experimental model. Animals were killed 3 months after transplant, and liver functions and histopathology were assessed. Results: Compared with the cirrhotic control group, the 3 cell-treated groups showed improved alanine aminotransferase, aspartate aminotransferase, albumin, and bilirubin levels, with best results shown in the hepatocyte-treated group. Histopathologic examination of the treated groups showed improved fibrosis, with best results obtained in the undif ferentiated mesenchymal stem cell-treated group. Conclusions: Both adult hepatocytes and cord blood mesenchymal stem cells proved to be promising candidates for cell-based therapy in liver regeneration on an experimental level. Improved liver function was evident in the hepatocyte-treated group, and fibrosis control was more evident in the undifferentiated mesenchymal stem cell-treated group.
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