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Host defense antimicrobial peptides (HDPs) constitute an integral component of the innate immune system having nonspecific activity against a broad spectrum of microorganisms. They also have diverse biological functions in wound healing, angiogenesis, and immunomodulation, where it has also been demonstrated that they have a high affinity to interact with human lipid signaling molecules. Within bacterial biofilms, quorum sensing (QS), the vital bacterial cell-to-cell communication system, is maintained by similar diffusible small molecules which control phenotypic traits, virulence factors, biofilm formation, and dispersion. Efficient eradication of bacterial biofilms is of particular importance as these colonies greatly help individual cells to tolerate antibiotics and develop antimicrobial resistance. Regarding the antibacterial function, for several HDPs, including the human cathelicidin LL-37, affinity to eradicate biofilms can exceed their activity to kill individual bacteria. However, related underlying molecular mechanisms have not been explored yet. Here, we employed circular dichroism (CD) and UV/VIS spectroscopic analysis, which revealed that LL-37 exhibits QS signal affinity. This archetypal representative of HDPs interacts with the Pseudomonas quinolone signal (PQS) molecules, producing co-assemblies with peculiar optical activity. The binding of PQS onto the asymmetric peptide chains results in chiral supramolecular architectures consisting of helically disposed, J-aggregated molecules. Besides the well-known bacterial membrane disruption activity, our data propose a novel action mechanism of LL-37. As a specific case of the so-called quorum quenching, QS signal molecules captured by the peptide are sequestered inside co-assemblies, which may interfere with the microbial QS network helping to prevent and eradicate bacterial infections.

Host defense antimicrobial peptides (HDPs) constitute an integral component of the innate immune system having nonspecific activity against a broad spectrum of microorganisms. They also have diverse biological functions in wound healing, angiogenesis, and immunomodulation, where it has also been demonstrated that they have a high affinity to interact with human lipid signaling molecules. Within bacterial biofilms, quorum sensing (QS), the vital bacterial cell-to-cell communication system, is maintained by similar diffusible small molecules which control phenotypic traits, virulence factors, biofilm formation, and dispersion. Efficient eradication of bacterial biofilms is of particular importance as these colonies greatly help individual cells to tolerate antibiotics and develop antimicrobial resistance. Regarding the antibacterial function, for several HDPs, including the human cathelicidin LL-37, affinity to eradicate biofilms can exceed their activity to kill individual bacteria. However, related underlying molecular mechanisms have not been explored yet. Here, we employed circular dichroism (CD) and UV/VIS spectroscopic analysis, which revealed that LL-37 exhibits QS signal affinity. This archetypal representative of HDPs interacts with the Pseudomonas quinolone signal (PQS) molecules, producing co-assemblies with peculiar optical activity. The binding of PQS onto the asymmetric peptide chains results in chiral supramolecular architectures consisting of helically disposed, J-aggregated molecules. Besides the well-known bacterial membrane disruption activity, our data propose a novel action mechanism of LL-37. As a specific case of the so-called quorum quenching, QS signal molecules captured by the peptide are sequestered inside co-assemblies, which may interfere with the microbial QS network helping to prevent and eradicate bacterial infections.

Macromolecular associates, such as membraneless organelles or lipid-protein assemblies, provide a hydrophobic environment, i.e., a liquid protein phase (LP), where folding preferences can be drastically altered. LP as well as the associated phase change from water (W) is an intriguing phenomenon related to numerous biological processes and also possesses potential in nanotechnological applications. However, the energetic effects of a hydrophobic yet water-containing environment on protein folding are poorly understood. Here, we focus on small β-sheets, the key motifs of proteins, undergoing structural changes in liquid–liquid phase separation (LLPS) and also model the mechanism of energy-coupled unfolding, e.g., in proteases, during W → LP transition. Due to the importance of the accurate description for hydrogen bonding patterns, the employed models were studied by using quantum mechanical calculations. The results demonstrate that unfolding is energetically less favored in LP by ~0.3–0.5 kcal·mol−1 per residue in which the difference further increased by the presence of explicit structural water molecules, where the folded state was preferred by ~1.2–2.3 kcal·mol−1 per residue relative to that in W. Energetics at the LP/W interfaces was also addressed by theoretical isodesmic reactions. While the models predict folded state preference in LP, the unfolding from LP to W renders the process highly favorable since the unfolded end state has &gt, 1 kcal·mol−1 per residue excess stabilization.

Silica@zirconia core@shell oxide nanoparticles of approx. 50 nm diameter were developed in view of an application as a novel carrier with adjuvant activities for cancer vaccines. To this end, the aim was to cover the surface of the nanocarrier by potent single stranded DNA (ssDNA) type immune stimulators used in cancer vaccination. Prior to characterizing the binding of ssDNA to the nanocarrier surface, the adsorption of deoxynucleoside monophosphate building blocks (dNMPs) was studied. After optimization of conditions for synthesis, solvent exchange and surface modification, the effects and possible interference of several buffers, such as HEPES, PIPES, MOPS and MOPSO, were also investigated. Formation of a new compound was revealed by UV–VIS spectroscopy during the recording of zeta potential vs. pH titration curves in the presence of HEPES and PIPES buffer in the neutral-acidic pH region. We identified this new species as gold ions etched from the electrode of the zeta cuvette, stabilized by the buffers via chelation, which is then followed by their conversion into gold nanoparticles.

Stable unilamellar dipalmitoylphosphatidylcholine vesicles were produced by using oligo(malic acid) and cholesterol. Detailed physico-chemical characterization prove that by using oligo(malic acid) the substitution of PEGylated lipids for sterically stabilization comes possible. The polymer molecules cover the outer surface of spherical-shaped vesicles, and an asymmetrical composition occurs in the two leaflets of the phospholipid bilayer. The oligo(malic-acid) and cholesterol are enriched in the outer side assuring the stabilization of vesicles. Cholesterol plays an important role in the self-assembly of components as it makes the entering of oligomers possible deep into the polar head-region of lipids. The presence of oligo(malic acid) molecules does not induce degradation by hydrolysis of lipid molecules but the vesicle system turns into a sensitive form giving a possibility for pH sensitive targeting. Preliminary investigation on the investigated oligo(malic acid)-stabilized vesicles do not show any toxic effect promising their applicability in the field of liposomal drug delivery.

The protein mediated approach is a common method for the synthesis of photoluminescent gold quantum clusters (GQCs), where proteins, acting as reducing and stabilizing agents, react with gold salts through cysteine residues. For the better understanding of the phenomenon, the aqueous phase reaction of HAuCl4 and l -cysteine has been investigated at the supramolecular level by various experimental techniques and molecular mechanics simulations. We have observed the formation of a novel photoluminescent product, ( AuCys ) n β , which shows emission in the orange region of the spectrum. Small- and wide-angle X-ray scattering (SWAXS) measurements have revealed the presence of nanosized lamellae, which have an internal multilayer superlattice structure with a characteristic periodic distance of 1.3 nm. Based on the results, the layers are built up by zigzag shaped (AuCys)n polymer chains connected through aurophilic bonds. The aurophilic network is stabilized via salt bridges and hydrogen bonds, which are also responsible for the interlayer interactions. Here, the evolution of the multilayer structure has been monitored by the combined application of photoluminescence spectroscopy and time-resolved SAXS. It has been concluded that there is a strong correlation between the emission and the scattering intensity, which suggests that the two- and three-dimensional aggregation of the building blocks to form sheets and multilayers are simultaneous processes. Furthermore, we have revealed that the formation and behavior of ( AuCys ) n β show significant differences to that of Au- l -glutathione compounds described earlier despite the similarity of l -cysteine and l -glutathione. These results evidence that l -cysteine and gold species form building blocks that can be applied expansively in supramolecular and cluster chemistry.

A new, simple, and “green” method was developed for the surface modification of 20 nm diameter Stober silica particles with 3-aminopropyl(diethoxy)methylsilane in ethanol. The bulk polycondensation of the reagent was inhibited and the stability of the sol preserved by adding a small amount of glacial acetic acid after appropriate reaction time. Centrifugation, ultrafiltration, and dialysis were compared in order to choose a convenient purification technique that allows the separation of unreacted silylating agent from the nanoparticles without destabilizing the sol. The exchange of the solvent to acidic water during the purification yielded a stable colloid, as well. Structural and morphological analysis of the obtained aminopropyl silica was performed using transmission electron microscopy (TEM), dynamic light scattering (DLS) and zeta potential measurements, Fourier-transform infrared (FTIR), 13 C and 29 Si MAS nuclear magnetic resonance (NMR) spectroscopies, as well as small angle X-ray scattering (SAXS). Our investigations revealed that the silica nanoparticle surfaces were partially covered with aminopropyl groups, and multilayer adsorption followed by polycondensation of the silylating reagent was successfully avoided. The resulting stable aminopropyl silica sol (ethanolic or aqueous) is suitable for biomedical uses due to its purity.

Extracellular vesicles isolated by differential centrifugation from Jurkat T-cell line were investigated by attenuated total reflection Fourier-transform infrared spectroscopy (ATR-FTIR). Amide and C H stretching band intensity ratios calculated from IR bands, characteristic of protein and lipid components, proved to be distinctive for the different extracellular vesicle subpopulations. This proposed ‘spectroscopic protein-to-lipid ratio’, combined with the outlined spectrum-analysis protocol is valid also for low sample concentrations (0.15–0.05 mg/ml total protein content) and can carry information about the presence of other non-vesicular formations such as aggregated proteins, lipoproteins and immune complexes. Detailed analysis of IR data reveals compositional changes of extracellular vesicles subpopulations: second derivative spectra suggest changes in protein composition from parent cell towards exosomes favoring proteins with β-turns and unordered motifs at the expense of intermolecular β-sheet structures. The IR-based protein-to-lipid assessment protocol was tested also for red blood cell derived microvesicles for which similar values were obtained. The potential applicability of this technique for fast and efficient characterization of vesicular components is high as the investigated samples require no further preparations and all the different molecular species can be determined in the same sample. The results indicate that ATR-FTIR measurements provide a simple and reproducible method for the screening of extracellular vesicle preparations. It is hoped that this sophisticated technique will have further impact in extracellular vesicle research.

Carbon nanotubes are promising new tools in biomedicine but they may have yet some unknown influences on the organism. In the present study, the acute effect of solubilized, multi-walled carbon nanotubes (MWCNTs) on basic neuronal functions was examined. Rat brain slices were treated in vitro with nanotube-containing colloid solutions at concentrations of 100-800 μg/ml and evoked field potentials were recorded from the somatosensory cortex and hippocampus. Basic excitability of the treated slices was characterized by the amplitude of field excitatory postsynaptic potentials (fEPSPs) and population spikes. Experimental results indicated significantly higher excitability of treated samples than that of controls. Multiple components in evoked potentials were observed, which is in accordance with the increased excitability of investigated brain areas. Tests of short- and long-term plasticity were also performed, which revealed no difference between control and treated slices. Experimental results suggest an interaction between nanotubes and brain tissue. MWCNTs seem to act on the basic membrane potential of neurons by changing membrane properties or via a mechanism linked to voltage-gated ion channels, rather than influencing specific synaptic transmission. Further investigation is needed to clarify the nature of interactions between nanotubes and brain tissue.