Inactivation of avian influenza viruses by hydrostatic pressure as a potential vaccine development approach.

Vaccines are a recommended strategy for controlling influenza A infections in humans and animals. Here, we describe the effects of hydrostatic pressure on the structure, morphology and functional characteristics of avian influenza A H3N8 virus. The effect of hydrostatic pressure for 3 h on H3N8 virus revealed that the particles were resistant to this condition, and the virus displayed only a discrete conformational change. We found that pressure of 3 kbar applied for 6 h was able to inhibit haemagglutination and infectivity while virus replication was no longer observed, suggesting that full virus inactivation occurred at this point. However, the neuraminidase activity was not affected at this approach suggesting the maintenance of neutralizing antibody epitopes in this key antigen. Our data bring important information for the area of structural virology of enveloped particles and support the idea of applying pressure-induced inactivation as a tool for vaccine production.

Trifluoperazine protects brain plasma membrane Ca(2+)-ATPase from oxidative damaging.

In the central nervous system (CNS), a number of different pathological processes such as necrosis, Parkinson's and Alzheimer's diseases are related to disturbance in calcium homeostasis associated with oxidative stress. Here we compare the susceptibility of rat brain plasma membrane Ca(2+)-ATPase (PMCA) and sarco-endoplasmic reticulum Ca(2+)-ATPase (SERCA) isoforms to in vitro oxidative stress, and investigate a putative role of trifluoperazine (TFP), an antipsychotic drug that is also a powerful inhibitor of Ca(2+)-transporter proteins, in protecting these enzymes. It is shown that, in rat brain, PMCA is very sensitive to the damage induced by preincubation with Fe(2+)-ascorbate, or Fe(2+)-ascorbate plus H2O2, while SERCA is resistant. Inhibition of PMCA activity promoted by Fe(2+)/ascorbate medium is fully prevented by the presence of microM concentrations of either butylated hydroxytoluene (BHT) or TFP, but only partially protected, or reversed, by dithiothreitol (DTT), pointing to some protein cysteine(s) as one of the main targets for a lipid peroxidation-dependent damaging mechanism. However, when 0.5-1 mM H2O2 is added together with Fe(2+)/ascorbate, both BHT and TFP only partially prevent ATPase activity inhibition, and DTT does not confer any protection, suggesting two possible additional mechanisms involving both lipid peroxidation and direct damage to PMCA at amino acid residues other than cysteines. A possible use of micromolar concentrations of TFP as a direct antioxidant protector for PMCA under oxidative stress conditions is discussed.