HCV is a hepatotropic RNA virus recognized for its frequent virulence and fatality worldwide. Despite many vaccine development programs underway, researchers are on a quest for natural bioactive compounds due to their multivalent efficiencies against viral infections, considering which the current research aimed to figure out the target-specificity and therapeutic potentiality of α, β, and δ subunits of amyrin, as novel bioactive components against the HCV influx mechanism. Initially, the novelty of amyrin subunits was conducted from 203 pharmacophores, comparing their in-silico pharmacokinetic and pharmacodynamic profiles. Besides, the best active site of CD81 was determined following the quantum tunneling algorithm. The molecular dynamic simulation was conducted (100 ns) following the molecular docking steps to reveal the parameters- RMSD (Å); Cα; RMSF (Å); MolSA (Å 2 ); Rg (nm); PSA (Å); SASA (Å 2 ), and the MM-GBSA dG binding scores. Besides, molecular strings of CD81, along with the co-expressed genes, were classified, as responsible for encoding CD81-mediated protein clusters during HCV infection, resulting in the potentiality of amyrins as targeted prophylactics in HCV infection. Finally, in vivo profiling of the oxidative stress marker, liver-specific enzymes, and antioxidant markers was conducted in the DMN-induced mice model, where β -amyrin scored the most significant values in all aspects.
Objectives: This research aimed to isolate, identify, and characterize a new strain of Bacillus cereus through different molecular biology approaches so that it could be further studied for therapeutic purposes against selective enteric pathogens. Materials and Methods: Pure isolates of B. cereus were prepared from buffalo yogurt samples in REMBA medium. Initially, the morphological, physiological, and biochemical properties were studied accordingly. Following the tests, the molecular identification for the strain identification was conducted through plasmid DNA extraction, PCR, agarose gel electrophoresis, and 16S rRNA sequencing up to 1.37 kb. Afterward, the antibiotic sensitivity [Epsilometer test (E-Test)] and anti¬fungal activity were tested considering different concentrations. Being classified from the aforementioned tests, a comprehensive antimicrobial activity test was conducted using the cell-free-su¬pernatant (CFS) of the test strain against selective enteric pathogens in humans in vitro. Besides, the different clusters of genes were identified and characterized for understanding the presump¬tive bacteriocins present in the CFS of the strain in silico, where molecular string properties were calculated. Finally, the evolutionary relationship among diversified bacteriocins synthesized by different Bacillus strains was studied to predict the CFS-containing bacteriocins of the new strain. Results: Purified isolates of B. cereus were Gram-positive rods and showed significant tolerance (p < 0.0001) to different concentrations of pH, phenol, bile salt, and NaCl. 16S rRNA revealed the strain as LOCK 1002, which was strongly sensitive to all the antibiotics used and resistant to selec¬tive antifungal agents. The CFS of B. cereus LOCK 1002 was found to be a very promising antago¬nist to all the enteric pathogens used in the culture condition. Two gene clusters were predicted to be interconnected and responsible for different presumptive bacteriocins. Conclusion: The newly identified LOCK 1002 can be a very potent strain of B. cereus in use as an antimicrobial agent for having different bacteriocin coding gene clusters.
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