To approve a theoretical basis for the molecular pathogenesis of human cerebral malaria and treatment with prevention Methods The blood samples were collected from 24 patients with cerebral malaria, 143 with falcipa...To approve a theoretical basis for the molecular pathogenesis of human cerebral malaria and treatment with prevention Methods The blood samples were collected from 24 patients with cerebral malaria, 143 with falciparum malaria, 34 with vivax malaria and 20 healthy controls from the endemic areas of Yunnan Province, China Using the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) technique, we determined the molecular mass (Mr) of these Plasmodium falciparum (P falciparum) erythrocyte membrane protein 1 (PfEMP1) molecules Results Our findings indicate that higher molecular mass (260?kDa-320?kDa) forms of PfEMP1 were expressed on parasitized erythrocyte (PE) from human cerebral malaria patients Compared with PfEMP1 expressed on PE from human cerebral malaria patients, the expression of PfEMP1 and Plasmodium vivax (P vivax) erythrocyte membrane protein 1 (PvEMP1) on PE from falciparum malaria patients and vivax malaria patients did not have multiple bands of PfEMP1 of ≥260?kDa, but had a PfEMP1 with molecular mass of 240?kDa and a PvEMP1 with molecular mass of 180?kDa band separately Healthy controls expressed an EMP of molecular mass of 140?kDa Conclusion Results confirm the antigenic variation of higher molecular mass of PfEMP1 whose molecular mass is equal to or exceeds 260?kDa-320?kDa on PE of patients with cerebral malaria Our results show that the binding of large antigenic variability PfEMP1 molecular mass of 260?kDa-320?kDa on PE from human cerebral malaria patients with diverse receptor molecules on the endothelial cell (EC) of the cerebral microvessels may be involved in the molecular pathogenesis of cerebral malaria展开更多
Objective:To isolate and characterize RNA aptamers that are specific to human CD36 protein using systematic evolution of ligands by exponential enrichment(SELEX)technology to identify candidates for adjunct therapy to...Objective:To isolate and characterize RNA aptamers that are specific to human CD36 protein using systematic evolution of ligands by exponential enrichment(SELEX)technology to identify candidates for adjunct therapy to reverse the binding of Plasmodiuminfected erythrocytes.Methods:RNA aptamers were isolated using nitrocellulose membrane-based SELEX and binding analysis was screened using an electrophoretic mobility shift assay and enzyme-linked oligonucleotide assay.Results:Thirteen cycles of nitrocellulose membrane-based SELEX yielded three aptamers(RC60,RC25,RC04)exhibiting high binding against CD36 protein as shown on electrophoretic mobility shift assay.The sequence analysis revealed a G-quadruplex sequence within all the isolated aptamers that might contribute to aptamer binding and thermodynamic stability.The specificity assay further showed that RC60 and RC25 were highly specific to CD36.The competitive inhibition assay demonstrated that RC60 and RC25 shared a similar binding epitope recognized by m Ab FA6-152,a specific monoclonal antibody against CD36.Conclusions:RC60 and RC25 are promising candidates as anticytoadherence for severe malaria adjunct therapy.展开更多
文摘To approve a theoretical basis for the molecular pathogenesis of human cerebral malaria and treatment with prevention Methods The blood samples were collected from 24 patients with cerebral malaria, 143 with falciparum malaria, 34 with vivax malaria and 20 healthy controls from the endemic areas of Yunnan Province, China Using the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) technique, we determined the molecular mass (Mr) of these Plasmodium falciparum (P falciparum) erythrocyte membrane protein 1 (PfEMP1) molecules Results Our findings indicate that higher molecular mass (260?kDa-320?kDa) forms of PfEMP1 were expressed on parasitized erythrocyte (PE) from human cerebral malaria patients Compared with PfEMP1 expressed on PE from human cerebral malaria patients, the expression of PfEMP1 and Plasmodium vivax (P vivax) erythrocyte membrane protein 1 (PvEMP1) on PE from falciparum malaria patients and vivax malaria patients did not have multiple bands of PfEMP1 of ≥260?kDa, but had a PfEMP1 with molecular mass of 240?kDa and a PvEMP1 with molecular mass of 180?kDa band separately Healthy controls expressed an EMP of molecular mass of 140?kDa Conclusion Results confirm the antigenic variation of higher molecular mass of PfEMP1 whose molecular mass is equal to or exceeds 260?kDa-320?kDa on PE of patients with cerebral malaria Our results show that the binding of large antigenic variability PfEMP1 molecular mass of 260?kDa-320?kDa on PE from human cerebral malaria patients with diverse receptor molecules on the endothelial cell (EC) of the cerebral microvessels may be involved in the molecular pathogenesis of cerebral malaria
基金supported by an Exploratory Research Grant Scheme(ERGS203/CIPPM/6730106)+2 种基金Higher Institution Centre of Excellent(HICo E311/CIPPM/4401005)from the Malaysian,Ministry of Higher EducationUniversiti Sains Malaysia Fellowship Scheme.
文摘Objective:To isolate and characterize RNA aptamers that are specific to human CD36 protein using systematic evolution of ligands by exponential enrichment(SELEX)technology to identify candidates for adjunct therapy to reverse the binding of Plasmodiuminfected erythrocytes.Methods:RNA aptamers were isolated using nitrocellulose membrane-based SELEX and binding analysis was screened using an electrophoretic mobility shift assay and enzyme-linked oligonucleotide assay.Results:Thirteen cycles of nitrocellulose membrane-based SELEX yielded three aptamers(RC60,RC25,RC04)exhibiting high binding against CD36 protein as shown on electrophoretic mobility shift assay.The sequence analysis revealed a G-quadruplex sequence within all the isolated aptamers that might contribute to aptamer binding and thermodynamic stability.The specificity assay further showed that RC60 and RC25 were highly specific to CD36.The competitive inhibition assay demonstrated that RC60 and RC25 shared a similar binding epitope recognized by m Ab FA6-152,a specific monoclonal antibody against CD36.Conclusions:RC60 and RC25 are promising candidates as anticytoadherence for severe malaria adjunct therapy.
