Lipid composition in fungal membrane models: effect of lipid fluidity.

The creation of effective fungal membrane models for neutron and X-ray reflectometry experiments is a key step in the development of new antifungal pharmaceuticals and agrochemicals to allow in vitro investigation of their mode of interaction with target cells. The structure of the obtained models depends on the properties of the lipids used and the final composition of the leaflets, and can be subject to the spontaneous translocation of phospholipids across the bilayer. The effect of phospholipid acyl-chain unsaturation and the presence of steroids in the membrane on the bilayer asymmetry were examined by means of neutron reflectometry. The measurements showed that membrane stability was higher if a zwitterionic, saturated acyl-chain phospholipid is present as the inner leaflet. Furthermore, membrane asymmetry was higher in the case of fully saturated lipid systems. As a result, membrane models consisting of fully saturated acyl chains within the inner leaflet are recommended as the starting point for subsequent studies of antifungal interactions owing to the simplicity of the models and their relative stability, thus allowing better control over the exact lipid composition facing the tested antifungal.

Biophysical studies in polymer therapeutics: the interactions of anionic and cationic PAMAM dendrimers with lipid monolayers.

Understanding how polymers interact with biological membranes is important for the development of polymer-based therapeutics and wider biomedical applications. Here, biophysical methods (surface pressure measurements, external reflection FTIR) have been used to investigate the interaction between PAMAM dendrimers (Generation 5 or 4.5) and anionic (DPPG) or zwitterionic (DPPC) model membranes. We observed a concentration-dependent binding behaviour of both PAMAM species to both model membranes; however, equivalent levels of penetration into DPPC monolayers required approximately 10-fold higher dendrimer concentrations than for penetration into DPPG monolayers. Overall, the anionic PAMAM G4.5 showed a slightly better penetration ability which could be caused by repulsive forces towards the lipid layers. In comparison, increasing concentration of cationic PAMAM G5 leads to saturation of adsorption at the anionic lipid surface before penetration into the lipid layer likely driven by electrostatic attraction. Our studies also showed that physiologically relevant concentrations of sodium chloride (144 mM) decreased PAMAM penetration into DPPG monolayers but did not significantly affect the dendrimer-DPPC interaction. These results provide an insight into the mechanism of interaction between charged dendritic polymers with a lipid interface and show that the nature of such interactions are affected by lipid headgroup, dendrimer charge and solution salinity.

Enzymatically Triggered, Isothermally Responsive Polymers: Reprogramming Poly(oligoethylene glycols) To Respond to Phosphatase.

Polymers which can respond to externally applied stimuli have found much application in the biomedical field due to their (reversible) coil-globule transitions. Polymers displaying a lower critical solution temperature are the most commonly used, but for blood-borne (i.e., soluble) biomedical applications the application of heat is not always possible, nor practical. Here we report the design and synthesis of poly(oligoethylene glycol methacrylate)-based polymers whose cloud points are easily varied by alkaline phosphatase-mediated dephosphorylation. By fine-tuning the density of phosphate groups on the backbone, it was possible to induce an isothermal transition: A change in solubility triggered by removal of a small number of phosphate esters from the side chains activating the LCST-type response. As there was no temperature change involved, this serves as a model of a cell-instructed polymer response. Finally, it was found that both polymers were non cytotoxic against MCF-7 cells (at 1 mg·mL(-1)), which confirms promise for biomedical applications.

Seizure frequency in pilocarpine-treated rats is independent of circadian rhythm.

Pilocarpine-induced status epilepticus (SE) results in chronic spontaneous recurrent seizures resembling human temporal lobe epilepsy. In this and other experimental models, behaviorally monitored seizure frequency was suggested to vary in a circadian fashion, and to increase with time. We re-addressed those hypotheses using continuous video-electroencephalography (EEG) telemetry in rats with SE at 30 days of age. In 11 chronically epileptic animals monitored up to 300 days after SE in a fixed 12 h light/dark cycle, we found that seizure frequency did not correlate with circadian rhythm.