ISOLATION OF GLICOPROTEID FROM THE FIXED RABIES VIRUS, STRAIN «MOSCOW 3253», AND CONSTRUCTING OF DOT-IMMUNOASSAY DIAGNOSTICUM ON ITS BASIS

Described here are the results of glicoproteid isolation from the fixed rabies virus, strain «Moscow 3253», using non-ionic detergent with subsequent chromatographic purification. The obtained antigen was demonstrated to be applicable as immunoreagent for construction of diagnosticum, by means of conjugation with colloid gold nanoparticles. The diagnosticum is meant for detection of specific antibodies in immune sera of horsesproducers, and in the preparation of anti-rabies immunoglobulin, in dot-immunoassay

Role of the Staphylococcus aureus Extracellular Loop of GraS in Resistance to Distinct Human Defense Peptides in PMN and Invasive Cardiovascular infections

GraS is a membrane sensor in Staphylococcus aureus that induces mprF and dlItABCD expression to alter the surface positive charge upon exposure to cationic human defense peptides (HDPs). The sensing domain of GraS likely resides in the 9residue extracellular loop (EL). In this study, we assessed a hospital-acquired methicillin-resistant S. aureus (HA-MRSA) strain (COL) for the specific role of two distinct EL mutations: F38G (bulk) and D/35/37/41K (charged inversion). Activation of mprF by polymyxin B (PMB) was reduced in the D35/37/41K mutant versus the D35/37/41G mutant, correlating with reduced surface positive charge; in contrast, these effects were less prominent in the F38G mutant but still lower than those in the parent. These data indicated that both electrostatic charge and steric bulk of the EL of GraS influence induction of genes impacting HDP resistance. Using mprF expression as a readout, we confirmed GraS signaling was pH dependent, increasing as pH was lowered (from pH 7.5 down to pH 5.5). In contrast to PMB activation, reduction of mprF was comparable at pH 55 between the P38G and D35/37/41K point mutants, indicating a mechanistic divergence between GraS activation by acidic pH versus cationic peptides. Survival assays in human blood and purified polymorphonuclear leukocytes (PMNs) revealed lower survival of the D35/37/41K mutant versus the F38G mutant, with both being lower than that of the parent. Virulence studies in the rabbit endocarditis model mirrored whole blood and PMN killing assay data described above. Collectively, these data confirmed the importance of specific residues within the EL of GraS in conferring essential bacterial responses for MRSA survival in infections.N