ABSTRACT
Background & objectives: The diagnosis of scrub typhus (ST) is usually done using enzyme-linked immunosorbent assay (ELISA) due to its ease of performance and reading objectivity. The cut-off value for ELISA needs to be calculated for each geographical location as it depends on zonal endemicity of the disease. This study was, therefore, undertaken to calculate the pan-India cut-off for anti-Orientia tsutsugamushi (OT) immunoglobulin M (IgM) by ELISA. Methods: Samples from cases (cases of ST) and controls (voluntary, consenting, healthy adults) were collected by a network of 29 laboratories across India and tested for anti-OT IgM by immunofluorescence assay (IFA), the considered gold standard test. These samples were retested by ELISA for anti-OT IgM and their optical densities (ODs) were used for cut-off estimation by receiver operating characteristic (ROC) curve. Results: Anti-OT IgM ELISA ODs from 273 controls and 136 cases were used for the cut-off estimation. The ODs of the anti-OT IgM ELISA on healthy individuals and those of confirmed ST cases ranged from 0.1 to 0.75 and 0.5 to 4.718, respectively. ROC curve-based cut-off for ELISA was calculated as 0.554 at a sensitivity of 95.2 per cent and specificity of 95.1 per cent. A value of >1 was noted to have a specificity of 100 per cent in diagnosing ST. Interpretation & conclusions: The cut-off calculated for India was similar to the previous cut-off that was used until now.
ABSTRACT
Bridelia ferruginea Benth. (Fam. 'Euphorbiaceae) is known to possess potent anti-inflammatory activity. Here, we investigated its vasomodulatory effect, as anti-inflammatory therapy that beneficially impact the cardiovascular system. Extracts (Bf1, Bf-HA) and fraction (Bf2) of B. ferruginea (Bf), were prepared from the bark of Bf to study their vasomodulatory effect using rat aortic rings. The vasorelaxant effect of Bf1 and Bf2 was mediated by the activation of nitric oxide synthase/endothelial isoform (NOS3) as confirmed by EA.hy926 endothelial cells, real-time PCR and Western blotting. Mass spectral analysis of these extracts and fraction was performed to understand the profile of compounds present in them. Mass spectral analysis showed the presence of similar ions in both Bf1 and Bf2 while Bf-HA showed different patterns. Vasorelaxant effect of Bf1 and Bf2 in phenylephrine (PE) pre-contracted endothelium intact aortic rings was blocked significantly in the presence of both N-nitro-L-arginine methyl ester (L-NAME) or soluble guanylate cyclase inhibitor (1H-[1,2,4]oxadiazolo-[4,3-a]quinoxalin-1-one [ODQ]). However, cyclo-oxygenase (COX) inhibitor (indomethacin) did not exert any change. In contrast, Bf-HA significantly inhibited ACh-induced vasorelaxation, but had no effect on sodium nitroprusside (SNP)-mediated relaxation, thereby suggesting NOS inhibitory activity in the extract. Studies with Bf1 and Bf2 on EA.hy926 cells demonstrated NOS3 mediated nitric oxide (NO) generation. Purified fractions of Bf, thus possess vasorelaxant compounds, which remain to be identified.