The Antimicrobial Peptide Histatin-5 Causes a Spatially Restricted Disruption on the Candida albicans Surface, Allowing Rapid Entry of the Peptide into the Cytoplasm

Antimicrobial peptides play an important role in host defense against microbial pathogens. Their high cationic charge and strong amphipathic structure allow them to bind to the anionic microbial cell membrane and disrupt the membrane bilayer by forming pores or channels. In contrast to the classical pore-forming peptides, studies on histatin-5 (Hst-5) have suggested that the peptide is transported into the cytoplasm of Candida albicans in a non-lytic manner, and cytoplasmic Hst-5 exerts its candicidal activities on various intracellular targets, consistent with its weak amphipathic structure. To understand how Hst-5 is internalized, we investigated the localization of FITC-conjugated Hst-5. We find that Hst-5 is internalized into the vacuole through receptor-mediated endocytosis at low extracellular Hst-5 concentrations, whereas under higher physiological concentrations, Hst-5 is translocated into the cytoplasm through a mechanism that requires a high cationic charge on Hst-5. At intermediate concentrations, two cell populations with distinct Hst-5 localizations were observed. By cell sorting, we show that cells with vacuolar localization of Hst-5 survived, while none of the cells with cytoplasmic Hst-5 formed colonies. Surprisingly, extracellular Hst-5, upon cell surface binding, induces a perturbation on the cell surface, as visualized by an immediate and rapid internalization of Hst-5 and propidium iodide or rhodamine B into the cytoplasm from the site using time-lapse microscopy, and a concurrent rapid expansion of the vacuole. Thus, the formation of a spatially restricted site in the plasma membrane causes the initial injury to C. albicans and offers a mechanism for its internalization into the cytoplasm. Our study suggests that, unlike classical channel-forming antimicrobial peptides, action of Hst-5 requires an energized membrane and causes localized disruptions on the plasma membrane of the yeast. This mechanism of cell membrane disruption may provide species-specific killing with minimal damage to microflora and the host and may be used by many other antimicrobial peptides

Weathering and Antimicrobial Properties of Laminate and Powder Coatings Containing Silver Phosphate Glass Used as High-Touch Surfaces

Increasing the use of hygienic high-touch surfaces with antimicrobial properties in health care and public spaces is one way to hinder the spread of bacteria and infections. This study investigates the antimicrobial efficacy and surface reactivity of commercial laminate and powder coated surfaces treated with silver-doped phosphate glass as antimicrobial additive towards two model bacterial strains, Escherichia coli and Bacillus subtilis, in relation to surface weathering and repeated cleaning. High-touch conditions in indoor environments were simulated by different extents of pre-weathering (repeated daily cycles in relative humidity at constant temperature) and simplified fingerprint contact by depositing small droplets of artificial sweat. The results elucidate that the antimicrobial efficacy was highly bacteria dependent (Gram-positive or Gram-negative), not hampered by differences in surface weathering but influenced by the amount of silver-doped additive. No detectable amounts of silver were observed at the top surfaces, though silver was released into artificial sweat in concentrations a thousand times lower than regulatory threshold values stipulated for materials and polymers in food contact. Surface cleaning with an oxidizing chemical agent was more efficient in killing bacteria compared with an agent composed of biologically degradable constituents. Cleaning with the oxidizing agent resulted further in increased wettability and presence of residues on the surfaces, effects that were beneficial from an antimicrobial efficacy perspective.QC 20220707</p

A novel methodology to study antimicrobial properties of high-touch surfaces used for indoor hygiene applications-A study on Cu metal.

Metal-based high-touch surfaces used for indoor applications such as doorknobs, light switches, handles and desks need to remain their antimicrobial properties even when tarnished or degraded. A novel laboratory methodology of relevance for indoor atmospheric conditions and fingerprint contact has therefore been elaborated for combined studies of both tarnishing/corrosion and antimicrobial properties of such high-touch surfaces. Cu metal was used as a benchmark material. The protocol includes pre-tarnishing/corrosion of the high touch surface for different time periods in a climatic chamber at repeated dry/wet conditions and artificial sweat deposition followed by the introduction of bacteria onto the surfaces via artificial sweat droplets. This methodology provides a more realistic and reproducible approach compared with other reported procedures to determine the antimicrobial efficiency of high-touch surfaces. It provides further a possibility to link the antimicrobial characteristics to physical and chemical properties such as surface composition, chemical reactivity, tarnishing/corrosion, surface roughness and surface wettability. The results elucidate that bacteria interactions as well as differences in extent of tarnishing can alter the physical properties (e.g. surface wettability, surface roughness) as well as the extent of metal release. The results clearly elucidate the importance to consider changes in chemical and physical properties of indoor hygiene surfaces when assessing their antimicrobial properties

Comparison and Functionalization Study of Microemulsion-Prepared Magnetic Iron Oxide Nanoparticles

Magnetic iron oxide nanoparticles (MION) for protein binding and separation were obtained from water-in-oil (w/o) and oil-in-water (o/w) microemulsions. Characterization of the prepared nanoparticles have been performed by TEM, XRD, SQUID magnetometry, and BET. Microemulsion-prepared magnetic iron oxide nanoparticles (ME-MION) with sizes ranging from 2 to 10 nm were obtained. Study on the magnetic properties at 300 K shows a large increase of the magnetization ~35 emu/g for w/o-ME-MION with superparamagnetic behavior and nanoscale dimensions in comparison with o/w-ME-MION (10 emu/g) due to larger particle size and anisotropic property. Moringa oleifera coagulation protein (MOCP) bound w/o- and o/w-ME-MION showed an enhanced performance in terms of coagulation activity. A significant interaction between the magnetic nanoparticles and the protein can be described by changes in fluorescence emission spectra. Adsorbed protein from MOCP is still retaining its functionality even after binding to the nanoparticles, thus implying the extension of this technique for various applications.We are grateful for the financial support of the Swedish Research Council, Formas, as well as the Cost Action D43, Colloid and Interface Chemistry for Nanotechnology. M.S.-D. acknowledges NaNoTeCh, the National Nanotechnology Laboratory of Mexico, and Cesar Leyva (CIMAV, S.C.) for HRTEM/STEM measurements and assistance, and A.T.T. (CIMAV, S.C.) for ATR-IR measurements and assistance. Financial support by Ministerio de Ciencia e Innovación (MICINN Spain, grant number CTQ2008-01979) and Generalitat de Catalunya (Agaur, grant number 2009SGR-961) is also aknowledged. C.C. acknowledges financial support from Spanish Ministerio de Ciencia e Innovacion; MAT 2008-02542 and GR35/10-A-950247.Peer reviewe