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A large part of the global population, including both immunocompromised and healthy individuals, suffers from fungal infections. The majority of current drug candidates for treating fungal infections exhibit poor water solubility, which hampers their permeability through biological barriers and limits their bioavailability. Here, we fabricated pea protein isolate (PPI)/poly-(ethylene oxide) nanofibers (PPI/PEO nanofibers), with a high protein content [65% (w/w)] by waterborne electrospinning as an eco-friendly drug delivery system. X-ray diffraction results demonstrated that the solid-state properties of the individual components (PPI and PEO) were retained in the PPI/PEO nanofibers. We then encapsulated the poorly water-soluble drug, clotrimazole (CTZ), in the nanofibers (PPI/PEO/CTZ nanofibers), without heat treatment and/or use of an organic solvent or surfactant to presolubilize CTZ, generating an antifungal delivery system for topical administration. An in vitro study demonstrated that CTZ was successfully loaded in and released from the nanofibers. Additionally, the nanofibers were not toxic to HeLa cells. Finally, based on an antifungal disc agar diffusion study, CTZ-loaded nanofibers were shown to be effective against Candida albicans. The overall results demonstrate the potential of PPI-based nanofibers as a green platform for the generation of CTZ-loaded efficient drug delivery systems for antifungal treatment. [ABSTRACT FROM AUTHOR]

The interactions and structural organization of water molecules play a crucial role in a wide range of physical, chemical, and biological processes. The ability of water to form hydrogen bonds (H-bonds) underpins its unique properties and enables it to respond dynamically to various environmental factors. These interactions at the molecular level may affect vital processes like protein folding, enzyme activity, and cellular organization. The presence of solutes and spatial constraints can alter the H-bonding network of water, and these effects are ubiquitous in the biological environment. In this study, we analyzed the fluorescence of 2-acetyl-6-(dimethylamino)naphthalene (ACDAN) fluorescence emission in water solutions containing kosmotropic and chaotropic salts and in agar hydrogels. Recently, this dye has proven invaluable in studying water network structure and dynamics, as its fluorescence signal changes based on the local dielectric environment, revealing variations in the dipolar relaxation of water. Our results show that ACDAN spectral response correlates with the degree of water ordering, providing important insights into solute–water interactions and water dynamics in free and confined environments. [ABSTRACT FROM AUTHOR]

We describe a method to obtain the brightness and number of molecules at each pixel of an image stack obtained with a laser scanning microscope. The method is based on intensity fluctuations due to the diffusion of molecules in a pixel. For a detector operating in the analog mode, the variance must be proportional to the intensity. Once this constant has been calibrated, we use the ratio between the variance and the intensity to derive the particle brightness. Then, from the ratio of the intensity to the brightness we obtain the average number of particles in the pixel. We show that the method works with molecules in solution and that the results are comparable to those obtained with fluctuation correlation spectroscopy. We compare the results obtained with the detector operating in the analog and photon counting mode. Although the dynamic range of the detector operating in the photon counting mode is superior, the performance of the analog detector is acceptable under common experimental conditions. Since most commercial laser scanning microscopes operate in the analog mode, the calculation of brightness and number of particles can be applied to data obtained with these instruments, provided that the variance is proportional to the intensity. We demonstrate that the recovered brightness of mEGFP, independent of concentration, is similar whether measured in solution or in two different cell types. Furthermore, we distinguish between mobile and immobile components, and introduce a method to correct for slow variations in intensity.