Twenty-one hepatitis B virus (HBV) isolates from the state of Haryana (North India) were studied for genotype, subgenotype, serotype distribution and precore mutations. Assays of alanine aminotransminase (ALT) and HBeAg were performed on all samples. Genotypes, subgenotypes and serotypes were determined by amplification of pre-S1/S2 regions followed by RFLP and also by phylogenetic analysis of amplified products. Mutations were studied by amplification and sequencing of the precore region. Twenty-four percent of the samples had high ALT levels and 90% were HBeAg negative. It was observed that 90% of the samples were HBV D genotype, (subgenotype D1, serotype ayw2), 5% HBV A genotype (subgenotype A1, serotype adw2), and the remaining 5% were HBV E genotype (serotype ayw4). The subgenotype A1 was quite similar to the South African isolates. Phylogenetic analysis of the HBV isolates, based on the pre-S1/S2 gene sequences, confirmed genotype E. Amplification and sequencing of the precore region showed 1762^A–T and 1764^G–A mutations in 38 and 15% of the samples, respectively. 1809^T was observed in 5% of the cases under study. This is the first report of the genotype E of hepatitis B virus in the Indian population. Efforts are underway to amplify and sequence the full length of this genotype E isolate.

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Hepatitis B virus (HBV) isolates (21) from Punjab (North India) were studied for genotype distribution and precore/core promoter mutations. Assays of alanine aminotransaminase (ALT) and HBeAg were performed in all isolates. Genotypes were determined in all the samples by restriction fragment length polymorphism and the precore/core promoter mutations were studied by amplification and by direct sequencing of precore/core promoter region. Sixty-two percent of the isolates had higher ALT levels and 57% of the isolates were HBeAg negative. It was observed that 90% of the isolates were HBV D genotype (subgenotype D1 and D2) and 10% of the isolates were HBV A genotype (subgenotype A1). Amplification and sequencing of the precore/core promoter region showed

Abbreviations: OD, optical density; CFU, colony forming unit; DMSO, dimethyl sulphoxide; SD, standard deviation; ZOI, zone of inhibition MOJ Drug Des Develop Ther. 2018;2(2):67-77 67 AbstractLiterature have ample evidence which showed that plant based secondary metabolites have profound antibacterial effect against pathogenic bacteria, but information regarding how the effect was generated is lacking. In the present study, we examined the effect of secondary metabolites from O. sanctum both in vitro and in silico to decode the probable pathway of inhibition. Experiments were divided into two parts-Wet and Dry lab experiments. In wet lab experiments, K. pneumoniae was taken as representative bacteria to evaluate the antibacterial potential of secondary metabolites present in different organic extracts of O. sanctum using agar-well diffusion and XTT-colorimetric methods. In dry lab experiments, 5 important secondary metabolites viz. Apigenin, Carvacrol, Eugenol, Ocimarin and Rosmarinic acid were docked against 5 essential enzymes of bacteria like Succinate dehydrogenase, MurG, MurE, DNA adenine methylase and DNA primase to assess their antibacterial potential in silico using Auto Dock Vina (1). The enzyme-ligand interaction of docked complexes was further analyzed by Molecular Dynamics Simulation technique using GROMACS(4.6.6). As per wet lab results, methanolic extract was found to be the most promising extract having highest antibacterial potential, where at 20mg/ml concentration of the extract there was complete inhibition of bacterial growth. As per dry lab results, Apigenin and Rosmarinic acid were found to be the most successful secondary metabolites which strongly inhibited MurE, MurG and DNA adenine methylase enzymes in silico. As both these metabolites are present in methanolic extract of O. sanctum, hence may be responsible for bactericidal effect of O. sanctum. Citation: Pahal V, Devi U, Singh M, et al. Significance of apigenin and rosmarinic acid mediated inhibition pathway of MurG, MurE and DNA adenine methylase enzymes with antibacterial potential derived from the methanolic extract of Ocimum sanctum. MOJ Drug Des Develop Ther. 2018;2(2):67-77.

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