Expression Profiling Of Selected Micro Rnas Coupled With Molecular And Biochemical Analyses Of Asiaticosides In Centella Asiatica (L.)Urb In Vitro

Objective: Centella asiatica (L.) Urb from Umbelliferae is a potential source of secondary metabolites having immense medicinal value. Asiaticoside is the major therapeutic compound. In the present study, Identification of a possible relationship between concentration/transcript level expression of asiaticoside and concentrations of growth hormones at different growth stages was observed. The current study includes molecular and biochemical evaluation of stress generated in C. asiatica at different time intervals in vitro. Methods: The enhancement in auxin, cytokinin and final asiaticoside content were determined using immunoassay kits for auxin, cytokinin and HPLC analysis respectively. Transcript level expression at different growth phases was carried out using real-time RT-PCR. For isolation of stress-related miRNAs, reverse transcription of total RNA using miScript II RT Kit PCR System was carried out as per instructions. The differential expression of five selected miRNAs was done by Real-Time RT-PCR. The analysis of stress in vitro was done by quantification of Hydrogen Peroxide (H2O2), total phenolics and total antioxidants by H2O2 assay kit, total antioxidant assay kit and Folin Ciocalteau reagent respectively. The final asiaticoside content was determined by HPLC. Results: Differential expression of key genes involved in asiaticoside pathway showed significantly higher transcript expression, which is in correlation with the final asiaticoside content. The enhanced expression of miRNAs and the analysis of H2O2, total antioxidant capacity and total phenolics are suggestive of generation of oxidative stress under controlled conditions. Conclusion: The present study shows a direct correlation between oxidative stress and transcript/phytochemical estimation of asiaticoside content under in vitro conditions.

Molecular p prediction, docking studies and synthesis of 5-benzimidazole-1-Yl-methyl –[1,3,4] oxadiazole-2-thiol and their derivatives.

Molecular property is a complex balance of various structural features which determine whether a particular molecule is similar to the known drugs.These properties mainly hydrophobicity, molecular size,flexibility and presence of various pharmacophoric features influence the behavior of molecules in a living organism, including oral bioavailability. This investigation deals with the design and calculation of molecular properties, drug likeness, lipophilicity and solubility parameters of 5-Benzimidazole-1-yl-methyl-[1, 3, 4] oxadiazole-2-thiol and their derivatives using Osiris, mol inspiration ,Mol soft software’s, and ALOPGPS 2.1 program. The compounds followed the Lipinski ‘Rule of five’ for better bioavailability, were synthesized and characterized by IR, NMR, and mass spectral analysis. Furthermore, the binding conformations of these compounds for anti inflammatory activities were determined in silico docking. This is an energy optimization process concerned with the search of the lowest free energy binding mode of a ligand within a protein binding site and estimates the forces involved in the protein-ligand recognition, carried out in Mastro V 2011 in the active site of the cyclooxygenase-2 (COX-2) enzyme.

Impact of different housing structures on filarial transmission in rural areas of southern India.

The aim of the present investigation was to assess the filarial transmission levels in houses of different structure in rural areas of Andhra Pradesh, India. During this study, ecologically-similar households were selected for entomological study. The per-man-hour density (PMHD), infection and infectivity rates, were recorded in different ranges ie, 16.1 to 77.6, 0-31.2% and 0-5.6%, respectively.

A baseline study of rural Bancroftian filariasis in southern India.

Night mass blood surveys were carried out for parasitological evidence of Bancroftian filariasis in 45 rural areas belonging to 9 National Filaria Control Program (NFCP) zones of East Godavari and West Godavari districts of Andhra Pradesh, India during the period 1998 to 2001. Mf prevalence range between 2.9 to 10.2%, and mf intensities in 20 mm3 blood samples ranged from 1-281. The present study explains the trend of microfilaria dynamics in the rural population, where mass drug delivery has been implemented since 1997, and anti-larivicidal and adulticidal control measures have not been adopted.

Relative abundance of Culex quinquefasciatus (Diptera: Culicidae) with reference to infection and infectivity rate from the rural and urban areas of East and West Godavari districts of Andhra Pradesh, India.

This paper describes the seasonal abundance of Culex quinquefasciatus in the rural and urban areas of the East and West Godavari districts (EGDT and WGDT) of Andhra Pradesh, India. The per man-hour density (PMHD) was collected from seven units in EGDT and two units in WGDT, which comprised rural and urban areas. The highest infection and infectivity rates were found in the rural areas of Rajahmundry (43.6%) and Amalapuram (13.2%) respectively. In urban areas, the highest infection and infectivity rates were found in Rajahmundry: 7.5% and 3.6% respectively. There was considerable difference in the infection rate and infectivity rates between the rural areas and urban areas in each unit.

Role of heme in mitochondrial biogensis: Transcriptional and post-transcriptional regulation of the expression of Iso-l-cytochrome C gene during glucose repression-derepression in cells of Saccharomyces cerevisiae.

Exogenous addition of hemin to glucose-repressed cells of Saccharomyces cerevisiae restores the level of Iso-1-cytochrome C messengers to that observed in derepressed cells. In vitro transcription in isolated nuclei has shown a 4-fold stimulation in the synthesis of Iso-1-cytochrome C messengers in repressed but hemin-treated and derepressed cells compared to the repressed cells. Studies on in vitro transport of RNA from isolated nuclei have revealed that there is a 50% drop in the transport of total RNA from nuclei isolated from repressed but hemin-treated and derepressed cells when compared with the nuclei from repressed cells. However, under these conditions, there is an enhanced transport of translatable RNA. Hybridization analysis of the transported RNA using Iso-1-cytochrome C gene-specific probe has shown that there is preferential transport of Iso-1-cytochrome C messengers in repressed but hemin treated and derepressed cells.