Natural self-emulsifying reversible hybrid-hydrogel delivery (N'SERH) of tocopherol enhances bioavailability and modulates alcohol-induced reproductive toxicity in rats.

Alpha-tocopherol (α-Toc), an antioxidant vitamin, has been widely prescribing in the treatment of infertility, in spite of its limited oral bioavailability. The present study describes the enhanced bioavailability and efficacy of a novel 'natural self-emulsifying reversible hydrogel' (N'SERH)-based oral delivery form of α-Toc-rich sunflower oil (Tα-fen) using fenugreek galactomannan hydrogel scaffold (hybrid-FENUMATTM ). Tα-fen was characterised by FTIR, SEM, TEM and DLS as a hybrid-hydrogel powder. The bioavailability study on thirty (n = 30) male Sprague Dawley rats randomised into two groups indicated 4.84-fold increase in the oral bioavailability when the formulation was provided at 15 mg/kg b. wt. of α-Toc by oral gavage. The efficacy study on 24 animals randomised into four groups as control, ethanol treated (4 mg/kg b. wt.), ethanol+unformulated, UTα (15 mg/kg b. wt.) and ethanol+formulation, Tα-fen (15 mg/kg b. wt.) revealed significant improvement (*p < 0.05) and reversal of alcohol-induced reproductive toxicity as evident from the enhanced sperm count, motility and viability parameters, testosterone levels, fructose content, and SDH activity and plasma antioxidant status among Tα-fen-treated rats, compared with unformulated, UTα-treated group. Histopathology further confirmed the reversal of the alterations in the testes morphology of Tα-fen-treated animals, indicating its promising potential in the treatment of reproductive health issues.

Cellular calcium signaling in the aging brain.

Aging in the biological system is an irreversible process. In the initial stages of lifespan aging improves survival skills of an organism while in the later stages aging reduce the survival skills. Aging is associated with changes in several cellular and molecular functions among which calcium signaling is a prominent one. Calcium signaling is essential for many vital functions of the brain and even minor impairments in calcium signaling can lead to deleterious consequences including neuronal death. Calcium signaling proteins are pursued as promising drug targets for many aging related diseases. This review attempts to summarize changes in calcium signaling in the brain as a result of aging.

Molecular dynamics simulations of the catalytic pathway of a cysteine protease: a combined QM/MM study of human cathepsin K.

Molecular dynamics simulations using a combined QM/MM potential have been performed to study the catalytic mechanism of human cathepsin K, a member of the papain family of cysteine proteases. We have determined the two-dimensional free energy surfaces of both acylation and deacylation steps to characterize the reaction mechanism. These free energy profiles show that the acylation step is rate limiting with a barrier height of 19.8 kcal/mol in human cathepsin K and of 29.3 kcal/mol in aqueous solution. The free energy of activation for the deacylation step is 16.7 kcal/mol in cathepsin K and 17.8 kcal/mol in aqueous solution. The reduction of free energy barrier is achieved by stabilization of the oxyanion in the transition state. Interestingly, although the "oxyanion hole" has been formed in the Michaelis complex, the amide units do not donate hydrogen bonds directly to the carbonyl oxygen of the substrate, but they stabilize the thiolate anion nucleophile. Hydrogen-bonding interactions are induced as the substrate amide group approaches the nucleophile, moving more than 2 A and placing the oxyanion in contact with Gln19 and the backbone amide of Cys25. The hydrolysis of peptide substrate shares a common mechanism both for the catalyzed reaction in human cathepsin K and for the uncatalyzed reaction in water. Overall, the nucleophilic attack by Cys25 thiolate and the proton-transfer reaction from His162 to the amide nitrogen are highly coupled, whereas a tetrahedral intermediate is formed along the nucleophilic reaction pathway.

Dynamics of an enzymatic substitution reaction in haloalkane dehalogenase.

Reactive flux molecular dynamics simulations have been carried out using a combined QM/MM potential to study the dynamics of the nucleophilic substitution reaction of dichloroethane by a carboxylate group in haloalkane dehalogenase and in water. We found that protein dynamics accelerates the reaction rate by a factor of 2 over the uncatalyzed reaction. Compared to the thermodynamic effect in barrier reduction, protein dynamic contribution is relatively small. However, analyses of the friction kernel reveal that the origins of the reaction dynamics in water and in the enzyme are different. In aqueous solution, there is significant electrostatic solvation effect, which is reflected by the slow reorganization relaxation of the solvent. On the other hand, there is no strong electrostatic coupling in the enzyme and the major effect on reaction coordinate motion is intramolecular energy relaxation.

Combined QM/MM study of the mechanism and kinetic isotope effect of the nucleophilic substitution reaction in haloalkane dehalogenase.

Combined QM/MM molecular dynamics simulations have been carried out for the dehalogenation reaction of the nucleophilic displacement of dichloroethane catalyzed by haloalkane dehalogenase. The computed chlorine kinetic isotope effects and free energies of activation in the wild-type and the Phe172Trp mutant enzyme are found to be consistent with experiment. In comparison with the uncatalyzed model reaction in water, the enzyme lowers the activation barrier by about 16 kcal/mol. The enormous enzymatic action was attributed to a combination of contributions from a change in the solvation effect and transition state stabilization. The unique features of tryptophan's ability to interact favorably with hydrophobic substrates and to form hydrogen bonds to the leaving group chloride ion at the transition state enable both factors to make significant contributions to the barrier lowering mechanism in the enzyme. This is in contrast to the reference reaction in water, in which hydrogen bonding interactions are weakened at the transition state because of dispersed charge distribution at the transition state relative to that in the reactant and product states.