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OBJECTIVE To study the bioactive phytochemicals in the leaves of A.ilicifolius against Hepatocellular carcinoma (HCC) through in silico, in vitro and in vivo studies. METHODS A. ilicifolius leaves were collected from Cuddalore District,Tamil Nadu,India.Authenticated by the Botanical Survey of India. The fresh leaves of A. ilicifolius were washed and shade dried at room temperature (28 ± 2)℃. The dried leaves were powdered by electrical blender.25 gms of A.ilicifolius leaf powder was used for methanol extraction in the Soxhlet apparatus.The Phytochemical compounds were analyzed by GC-MS and the structure was retrieved from PubChem.Totally,seven HCC target proteins were collected from literature, ligand and proteins were prepared for in silico molecular docking. HepG2 cell lines were used for in vitro (MTT assay). BALB/c mice were used for in vivo studies, the biochemical parameters and histopathological studies were carried out with standard procedure. RESULTS The phytochemical 26.27-Di (nor)-cholest-5, 7, 23-trien-22-ol, 3-methoxymethoxy exhibited maximum docking score against the HCC target protein C-Jun N-terminal kinase 1 (JNK 1) (-6.839798). MTT assay revealed that the extract at a concentration of 200 μg·mL-1,caused 60% cell death in HepG2 cell lines.Further animal studies using to injecting HCC induced BALB/c mice,restoration of haematological parameters and cells to normal was observed after 15 days of oral administration of the extract.These findings suggest the possibility of using A.ilicifolius leaves in the treatment of HCC.CONCLUSION The in silico,in vitro and in vivo studies indicated that the A.ilicifolius leaves had anticancereous activity against Hepatocellular carcinoma.There can be a possibility of synergistic activity of phytochemicals together against HCC.
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Mutations within transpeptidase domain of penicillin-binding protein 2B of the strains of Streptococcus pneumoniae leads to resistance against ß-lactam antibiotics. To uncover the important residues responsible for sensitivity and resistance, the recently determined three dimensional structures of penicillin-binding protein 2B of both wild-type R6 (sensitive) and mutant 5204 (resistant) strains along with the predicted structures of other mutant strains G54, Hungary19A-6 and SP195 were considered for the interaction study with ß-lactam antibiotics using induced-fit docking of Schrödinger. Associated binding energies of the complexes and their intermolecular interactions in the binding site clearly show that the wild-type R6 as sensitive, mutant strains 5204 and G54 as highly resistant, and the mutant strains Hungary19A-6 and SP195 as intermediate resistant. The study also reveals that the mutant strains Hungary19A-6 and SP195 exhibit intermediate resistant because of the existence of mutations till the intermediate 538th and 516th positions, respectively, and not till the end of the C-terminus. Furthermore, our investigations show that if the mutations are extended till the end of the C terminus, then the antibiotic resistance of induced-mutated strains increases from intermediate to high as in the strains 5204 and G54. The binding patterns obtained in the study are useful in designing potential inhibitors against multidrug resistant S. pneumoniae.