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Objective To establish a suitable method of diagnosis of visceral leishmaniasis (VL) using peripheral blood, spleen or bone marrow aspirates. Methods Peripheral blood, bone marrow and spleen aspirate samples were collected from clinically suspected VL patients (n = 26). A new PCR primer pair (MK1F/R) was designed targeting kinetoplast mini circle DNA sequences of Leishmania donovani, and Leishmania infantum, and was used to diagnose VL along with some other established primers for VL in polymerase chain reactions. Test was validated by comparing with several other diagnostic methods. Results The designed primer set showed 100% specificity and 98% sensitivity in detecting VL using blood samples, when compared with more invasive samples: bone marrow or spleen aspirates. Conclusions The newly designed primer MK1F/R could be a better alternative for PCR based diagnosis of VL using less invasive sample, peripheral blood instead of bone marrow or spleen aspirates.
RESUMO
OBJECTIVE@#To establish a suitable method of diagnosis of visceral leishmaniasis (VL) using peripheral blood, spleen or bone marrow aspirates.@*METHODS@#Peripheral blood, bone marrow and spleen aspirate samples were collected from clinically suspected VL patients (n = 26). A new PCR primer pair (MK1F/R) was designed targeting kinetoplast mini circle DNA sequences of Leishmania donovani, and Leishmania infantum, and was used to diagnose VL along with some other established primers for VL in polymerase chain reactions. Test was validated by comparing with several other diagnostic methods.@*RESULTS@#The designed primer set showed 100% specificity and 98% sensitivity in detecting VL using blood samples, when compared with more invasive samples: bone marrow or spleen aspirates.@*CONCLUSIONS@#The newly designed primer MK1F/R could be a better alternative for PCR based diagnosis of VL using less invasive sample, peripheral blood instead of bone marrow or spleen aspirates.
RESUMO
Present research work was designed to study the anticancer and antioxidant activities of methanol extract of Mussaenda roxburghii. Anticancer activity of MMR has been carried out on Ehrlich ascites carcinoma [EAC] cells with three different doses [20, 40 and 60 mg/kg/day] by observing different parameters such as tumor weight, survival time of EAC-bearing mice, growth inhibition of EAC cells, morphological changes and nuclear damage of EAC cells etc. whereas antioxidant activity was determined by measuring total antioxidant, DPPH free radical scavenging, ferrous reducing capacity assay. The extract showed highest anticancer activity at 60 mg/kg day[-1][i.p.]. It caused 81.4% [P<0.01] cells growth inhibition and reduced tumor burden significantly [78.5%; P<0.001] in comparison to control. It also increased life span of EAC-bearing mice significantly [73.5%; P<0.01]. MMR treated EAC cells showed membrane blebbing, chromatin condensation, nuclear fragmentation [apoptotic feature] in Hoechst 33342 staining under fluorescence microscope. DNA fragmentation assay in agarose gel [1.5%] electrophoresis also rectified that it causes EAC cells death by apoptosis. MMR also exhibited moderate antioxidant properties in dose dependent manner. Thus, this plant can therefore be considering a resource for natural chemo-preventive drugs as well as a possible pharmaceutical supplement