Carbapenems and Lipid Bilayers: Localization, Partitioning, and Energetics.

Carbapenems are broad-spectrum antibiotics used today to treat otherwise antibiotic resistant bacteria. As their target transpeptidase is located within the periplasm of the Gram-negative bacteria, they can participate in nonspecific interactions between the inner leaflet of the outer membrane and the outer leaflet of the inner membrane. We, therefore, studied the interaction of the four most clinically relevant carbapenems, namely, imipenem, doripenem, ertapenem, and meropenem, with model phospholipid bilayers made of 1,2-dimyristoyl- sn-glycero-3-phosphocholine (DMPC) using molecular dynamics (MD) simulations and X-ray diffraction at low and high concentration of the drugs corresponding to 1 and 8 mol % (with respect to the number of membrane lipids). Membrane solubility was found to decrease from imipenem to doripenem, ertapenem, and finally meropenem. At low concentrations, membrane insertion was found to be a two step process, where the drugs first adsorb to the lipid head groups before inserting through a rotation of the molecule. At higher drug concentrations, the molecules were found to form aggregates in the aqueous phase before making contact with the membranes and spontaneously inserting into the bilayers. Two populations of imipenem were found: in the headgroup at ∼17 Å from the bilayer center and an inserted population at z-values of about 7 Å. Other carbapenems were found to localize in the tail groups with meropenem at ∼10 Å, doripenem at ∼8 Å, and ertapenem at ∼8 Å. The observed membrane solubility of carbapenems can potentially impact the availability of the drug to the target penicillin-binding proteins, potentially affecting their clinical efficacy.

Bacterial Suspensions Deposited on Microbiological Filter Material for Rapid Laser-Induced Breakdown Spectroscopy Identification.

Four species of bacteria, E. coli, S. epidermidis, M. smegmatis, and P. aeruginosa, were harvested from agar nutrient medium growth plates and suspended in water to create liquid specimens for the testing of a new mounting protocol. Aliquots of 30 µL were deposited on standard nitrocellulose filter paper with a mean 0.45 µm pore size to create highly flat and uniform bacterial pads. The introduction of a laser-based lens-to-sample distance measuring device and a pair of matched off-axis parabolic reflectors for light collection improved both spectral reproducibility and the signal-to-noise ratio of optical emission spectra acquired from the bacterial pads by laser-induced breakdown spectroscopy. A discriminant function analysis and a partial least squares-discriminant analysis both showed improved sensitivity and specificity compared to previous mounting techniques. The behavior of the spectra as a function of suspension concentration and filter coverage was investigated, as was the effect on chemometric cell classification of sterilization via autoclaving.