Enhancing encapsulation of filarial antigen Brugia malayi thioredoxin in nano-liposomes: The role of lecithin composition.

Background and purpose: Lymphatic filariasis is a debilitating infectious disease prevalent in endemic areas, necessitating the development of an effective vaccine for eradication. Although recombinant vaccine candidates have been deemed safe, they often fail to provide sufficient protection, which can be overcome by encapsulating them in nano-liposomes. In this study, we have optimised the liposomal composition for enhanced stability and encapsulation of filarial antigen Brugia malayi thioredoxin (Bm-TRX). Experimental approach: Nano-liposomes were prepared using egg phosphatidylcholine (EPC) and cholesterol via thin-film hydration, followed by sonication and characterizing. Encapsulation efficiency was optimised using different weight ratios of EPC to cholesterol (8:2, 7:3, and 6:4) and total lipid (EPC+Cholesterol) concentration to antigen Bm-TRX (10:1, 10:2, and 10:3) followed by release kinetics study. Key results: Optimised parameters yielded spherical liposomes measuring 209 nm in diameter with narrow polydispersity. Our findings demonstrated the highest encapsulation efficiency of 70.685 % and stability of 10 hours for an EPC to cholesterol weight ratio of 7:3. The in silico study proved the antigenic nature of TRX. Conclusion: The liposomal formulations loaded with TRX, as optimized in this study, hold promise for improving antigen efficiency by enhancing stability, bioavailability, and prophylactic effects by acting as immune potentiators.

In-silico structural studies on anti-inflammatory activity of phytocompounds from the genus Andrographis.

Plant species from the genus Andrographis were used in Ayurveda and in other folklore medicines for treating ailments for centuries. In this study, we have hypothesized to evaluate the secondary metabolites as potent anti-inflammatory agents from the genus Andrographis. A library of secondary metabolites of the said genus was curated from the PubChem database and was subjected to energy minimization using UCSF Chimera software employing the AMBER force field. Initially, molecular docking was performed to evaluate the binding affinity of the curated library against the enzymes cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), and 5-Lipoxygenase (5-LOX) using AutoDock Vina. This resulted in shortlisting of two metabolites Echioidinin 5-O-glucoside was bound and 5,2',6'-Trihydroxy-6,7,8-trimethoxy flavone 2'-O-glucoside with high binding affinity than standard drugs Ibuprofen and Zileuton. In addition, molecular dynamic simulation studies confirm that these compounds form relatively more stable complexes than standard drugs with the above-said enzymes. The free binding energy values using MMGBSA of the above said ligands with COX-1, COX-2, and 5-LOX were found to be -49.18 kcal/mol, -38.60 kcal/mol, and -54.27 kcal/mol respectively, Whereas the standards have -21.77 kcal/mol, -9.96 kcal/mol, and -10.29 kcal/mol. Moreover, the in-silico ADMET analysis confirms the druggability of the shortlisted compounds. Later, this work will act as a base for in-vitro and in-vivo experimental studies to validate the anti-inflammatory potential of the same.Communicated by Ramaswamy H. Sarma.

An Insilico evaluation of phytocompounds from Albizia amara and Phyla nodiflora as cyclooxygenase-2 enzyme inhibitors.

PURPOSE: The enzyme Cyclooxygenases (COX-1 and COX-2) catalyze the formation of prostaglandin, a mediator of the inflammatory pathway. Inflammation related pathological conditions may be alleviated by targeting the Cox enzymes.COX-2 inhibitors that are currently available in the market causes undesirable side effects. Our present study focuses on the in-silico inhibition of COX -2 enzyme by the phytocompounds from Albizia amara and Phyla nodiflora. METHODS: The phytochemicals present in Albizia amara and Phyla nodiflora were analyzed for their COX-2 inhibition potential. Eight compounds from Albizia amara and eleven compounds from Phyla nodiflora obtained from GC-MS analysis was used for the current study. Molecular docking was performed using AutoDock vina. The crystal structure of COX-2 (PDB ID: 5IKR) was obtained from Protein data bank. PyMol was used to remove any solvent, organic and inorganic molecules. Energy minimization of the protein was carried out using SPDBV software. Geometrical optimizations of the ligands were performed using Avogadro software. Celecoxib was used as the positive control. ADMET properties of the compounds were analyzed using SwissADME and ProtoxII online servers. Molecular mechanics/generalized born surface area (MM/GBSA) calculations were performed to evaluate the binding efficiency. Molecular dynamics of the protein and protein-ligand complex was studied for about 100 ns using Desmond package of Schrodinger suite. RESULTS: Among the eighteen compounds, Squalene present in both the plants showed a better binding energy of -7.7 kcal/mol, when compare to other phytocompounds present in the extract. The control celecoxib showed a binding energy of about - 9.4 kcal/mol. The toxicity and ADMET properties of squalene indicated that it is non-toxic and followed Lipinski's rule. Molecular Dynamics (MD) analysis showed that the binding of squalene to the enzyme was stable. CONCLUSION: Squalene could potentially inhibit COX2 and o wing to its properties, squalene can be formulated in gels/creams and could be possibly used for external edema and inflammation.

Acid stable α-amylase from Pseudomonas balearica VITPS19-Production, purification and characterization.

In the present study, α-amylase from Pseudomonas balearica VITPS19 isolated from Kolathur, Tamil Nadu, India was studied. Initially, one factor at a time (OFAT) approach was used to optimize the medium parameters like pH, temperature, carbon and nitrogen sources and the presence of metal ions to enhance the amylase activity. After the optimization, 6.5-fold increase in the enzyme production was observed. Enzyme purification was carried out in three stages. The molecular weight of purified α-amylase was estimated to be 47 kDa.The optimum activity for the purified enzyme was observed at pH 6 in 0.1 M phosphate buffer at 25 ± 2 °C and the activity is enhanced in the presence of ions like Mn2+, Mo6+, Na+, Mg2+and Zn2+ and was inhibited in the presence of Hg2+ ions. Compounds such as Sodium dodecyl sulfate (SDS), Ethylenediaminetetraacetic acid (EDTA), urea and ß- mercaptoethanol reduced the amylase activity. The Km and Vmax of the α-amylase was estimated to be 45.23 mM and 20.83 U/mL, respectively.