Relative stability of human centrins and its relationship to calcium binding.

Centrins are calcium binding proteins that belong to the EF-hand superfamily with diverse biological functions. Herein we present the first systematic study that establishes the relative stability of related centrins via complementary biophysical techniques. Our results define the stepwise molecular behavior of human centrins by two-dimensional infrared (2D IR) correlation spectroscopy, the change in heat capacity and enthalpy of denaturation by differential scanning calorimetry, and the relative stability of the helical regions of centrins by circular dichroism. More importantly, 2D IR correlation spectroscopy provides unique information about the similarities and differences in dynamics between these related proteins. The thermally induced molecular behavior of human centrins can be used to predict biological target interactions that have a relative dependence on calcium affinity. This information is essential for understanding why certain isoforms may be used to rescue a phenotype and therefore also for explaining the different functions these proteins may have in vivo. Furthermore, this comparative approach can be applied to the study of recombinant therapeutic protein candidates for the treatment of disease states.

Prospective study using standardized methodology for antimicrobial susceptibility of gram-positive cocci isolated from the Puerto Rico Medical Center.

The Gram-positive cocci (GPC), Staphylococcus aureus, Streptococcus pneumoniae, Enterococcus faecalis and Enterococcus faecium, have become important causes of community and nosocomial-acquired infections. The prevalence of multiple resistant isolates to standard antimicrobial drugs has significantly increased over the past decades. Few prospective studies have been performed in Puerto Rico (PR) concerning the GPC antimicrobial susceptibilities pattern. The purpose of this study was to evaluate the in vitro susceptibility of GPC clinical isolates from PR to selected standard antibiotics and to the new antimicrobial agents, linezolid (LZ), quinupristin/dalfopristin (Q/D) and gemifloxacin (GM). The in vitro susceptibility utilizing disk diffusion and Etest methods to selected antibiotics was determined for a total of 429 isolates obtained during a period of 5 months from the Puerto Rico Medical Center Bacteriology Laboratory. The distribution of GPC collected was as follows: 213 S. aureus isolates, 162 E. faecalis, 16 E. faecium and 38 S. pneumoniae. The results of the susceptibility test demonstrated: 1) that in S. aureus, 100% of the isolates were susceptible to vancomycin (VAN), LZ and Q/D; 93% to GM; and 61% to methicillin/oxacillin; 2) in S. pneumoniae, 100% were susceptible to LN and GM; 87% to Q/D; and 53% to penicillin; 3) in E. faecalis, 99% were susceptible to ampicillin; 93% to LZ; 79% to GM; 78.6% to VAN; and 0% to Q/D. Sixty eight and 66% of the E. faecalis isolates were susceptible to gentamicin and streptomycin respectively; and 4) in E. faecium, 100% were susceptible to LZ; 94% to Q/D; 69% to GM; 37.5% to VAN and 20% to ampicillin. In E. faecium isolates, 50% and 31% were susceptible to gentamicin and streptomycin, respectively. Of the vancomycin resistant enterococci, 88.9% and 21% of E. faecium and faecalis showed VanA phenotypic resistance, respectively. These results show that there is a significant degree of antimicrobial resistance in GPC, including 38% methicillin resistance in S. aureus, a near 50% penicillin resistant S. pneumoniae, and a significant resistance of enterococcal species to VAN. The new agents, LZ, Q/D and GM, proved to be effective against both, S. aureus and S. pneumoniae. For E. faecium, both, LZ and Q/D were active, while for E. faecalis, only LZ showed consistent activity.