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We propose minimal transport experiments in the coherent regime that can probe the chirality of twisted moiré structures. We show that only with a third contact and in the presence of an in-plane magnetic field (or another time-reversal symmetry breaking effect) a chiral system may display nonreciprocal transport in the linear regime. We then propose to use the third lead as a voltage probe and show that opposite enantiomers give rise to different voltage drops on the third lead. Additionally, in the scenario of layer-discriminating contacts, the third lead can serve as a current probe capable of detecting different handedness even in the absence of a magnetic field. In a complementary configuration, applying opposite voltages on the two layers of the third lead gives rise to a chiral (super)current in the absence of a source-drain voltage whose direction is determined by its chirality.

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PURPOSE: This study evaluated the fatigue failure load (FFL) and the number of cycles for fatigue failure (CFF) of traditional (porcelain layer up) and reversed (zirconia layer up) designs of porcelain-veneered zirconia samples prepared with heat-pressing or file-splitting techniques. MATERIALS AND METHODS: Zirconia discs were prepared and veneered with heat-pressed or machined feldspathic ceramic. The bilayer discs were bonded onto a dentin-analog according to the bilayer technique and sample design: traditional heat-pressing (T-HP), reversed heat-pressing (R-HP), traditional file-splitting with fusion ceramic (T-FC), reversed file-splitting with fusion ceramic R-FC), traditional file-splitting with resin cement (T-RC), and reversed file-splitting with resin cement (R-RC). The fatigue tests were performed using the stepwise approach at 20 Hz, 10,000 cycles/step, step-size of 200 N starting at 600 N, and proceeding until failure detection or up to 2600 N if enduring. The failure modes (from radial and/or cone cracks) were analyzed in a stereomicroscope. RESULTS: The reversed design decreased the FFL and CFF of bilayers prepared with heat-pressing and file-splitting with fusion ceramic. The T-HP and T-FC reached the highest results, which were statistically similar between them. The bilayers prepared by the file-splitting with resin cement (T-RC and R-RC) were similar to the R-FC and R-HP groups regarding FFL and CFF. Almost all reverse layering samples failed by radial cracks. CONCLUSIONS: The reverse layering design did not improve the fatigue behavior of porcelain veneered zirconia samples. The three bilayer techniques behaved similarly when used in the reversed design.

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Bioactive peptides have emerged as promising therapeutic agents with antimicrobial, antifungal, antiparasitic, and, recently, antitumoral properties with a mechanism of action based on membrane destabilization and cell death, often involving a conformational change in the peptide. This biophysical study aims to provide preliminary insights into the membrane-level antitumoral mode of action of crotalicidin, a cationic host defense peptide from rattlesnake venom, toward breast cancer cell lines. The lipid composition of breast cancer cell lines was obtained after lipid extraction and quantification to prepare representative cell membrane models. Membrane-peptide interaction studies were performed using differential scanning calorimetry and Fourier-transform infrared spectroscopy. The outcome evidences the potential antitumoral activity and selectivity of crotalicidin toward breast cancer cell lines and suggests a mechanism initiated by the electrostatic interaction of the peptide with the lipid bilayer surface and posterior conformation change with membrane intercalation between the acyl chains in negatively charged lipid systems. This research provides valuable information that clears up the antitumoral mode of action of crotalicidin.

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PURPOSE: To evaluate the effect of different veneering ceramics and veneering techniques on the bond strength to zirconia. MATERIALS AND METHODS: 3Y-TZP zirconia blocks were sliced into 60 slabs, polished, and sintered. Each slab received one ceramic cylinder (Ø = 3.4 mm, 5 mm-high), according to the veneering ceramic type (feldspathic-FEL or lithium disilicate-based-LD) and the veneering technique (file-splitting with resin-based luting agent-RC, file-splitting with fusion ceramic-FC, or heat-pressing-HT), which resulted in six groups: FEL-RC, FEL-FC, FEL-HT, LD-RC, LD-FC, LD-HT. After preparation, the samples were immersed in distilled water for 24 h before the shear bond strength (SBS) test. The failure modes were classified as adhesive, predominantly adhesive, or cohesive. Representative failure mode images were taken in a Scanning Electron Microscope. The SBS data were analyzed by two-way ANOVA and Tukey's test. RESULTS: Both type of veneering ceramic and technique affected the bond strength. FC led to the highest SBS values. RC and HP provided similar results when compared within each veneering ceramic. Lithium disilicate achieved lower bond strength than feldspathic ceramic when the heat-pressing technique was applied. The most frequent failure modes were predominantly adhesive and adhesive for FEL and LD, respectively. CONCLUSION: File-splitting with fusion ceramic provided the highest adhesion to zirconia when feldspathic or lithium disilicate-based ceramics were used. The heat-pressing technique for veneering with lithium disilicate significantly decreased the bond strength when compared to the feldspathic ceramic.

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The plasma membrane (PM) has undergone important conceptual changes during the history of scientific research, although it is undoubtedly a cellular organelle that constitutes the first defining characteristic of cellular life. Throughout history, the contributions of countless scientists have been published, each one of them with an enriching contribution to the knowledge of the structure-location and function of each structural component of this organelle, as well as the interaction between these and other structures. The first published contributions on the plasmatic membrane were the transport through it followed by the description of the structure: lipid bilayer, associated proteins, carbohydrates bound to both macromolecules, association with the cytoskeleton and dynamics of these components.. The data obtained experimentally from each researcher were represented in graphic configurations, as a language that facilitates the understanding of cellular structures and processes. This paper presents a review of some of the concepts and models proposed about the plasma membrane, emphasizing the components, the structure, the interaction between them and the dynamics. The work is illustrated with resignified 3D diagrams to visualize the changes that occurred during the history of the study of this organelle. Schemes were redrawn in 3D from the original articles...

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Cationic and hydrophilic coatings based on casting and drying water dispersions of two different nanoparticles (NPs) onto glass are here described and evaluated for antimicrobial activity. Discoid cationic bilayer fragments (BF) surrounded by carboxy-methylcellulose (CMC) and poly (diallyl dimethyl ammonium) chloride (PDDA) NPs and spherical gramicidin D (Gr) NPs dispersed in water solution were cast onto glass coverslips and dried, forming a coating quantitatively evaluated against Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans. From plating and colony forming units (CFU) counting, all strains interacting for 1 h with the coatings lost viability from 105 to 106, to zero CFU, at two sets of Gr and PDDA doses: 4.6 and 25 µg, respectively, or, 0.94 and 5 µg, respectively. Combinations produced broad spectrum, antimicrobial coatings; PDDA electrostatically attached to the microbes damaging cell walls, allowing Gr NPs interaction with the cell membrane. This concerted action promoted optimal activity at low Gr and PDDA doses. Further washing and drying of the deposited dried coatings showed that they were washed out so that antimicrobial activity was no longer present on the glass surface. Significant applications in biomedical materials can be foreseen for these transient coatings.

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This paper describes the development of a coating for cotton and polypropylene (PP) fabrics based on a polymeric matrix embedded with cuprous oxide nanoparticles (Cu2O@SDS NPs) in order to inactivate SARS-CoV-2 and manufactured by a simple process using a dip-assisted layer-by-layer technology, at low curing temperature and without the need for expensive equipment, capable of achieving disinfection rates of up to 99%. The polymeric bilayer coating makes the surface of the fabrics hydrophilic, enabling the transportation of the virus-infected droplets to achieve the rapid inactivation of SARS-CoV-2 by contact with the Cu2O@SDS NPs incorporated in the coated fabrics.

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OBJECTIVES: To evaluate the longevity of metal-ceramic single crowns cemented onto resin composite prosthetic cores using a self-adhesive resin cement in a prospective clinical descriptive study. METHODS: A total of 152 teeth were endodontically treated and received resin composite prosthetic cores and metal-ceramic crowns cemented with a self-adhesive resin cement. The patients included in the sample were recalled for clinical and radiography evaluation in an up-to-106-month period after the final cementation procedures, with an average of 62 months of follow-up. 91.5% of the sample (142 teeth) were evaluated regarding the treatment survival rate, analyzed considering the loss of crown retention (crown debonding) and tooth loss as the primary outcome. In addition, post debonding, and root fracture occurrences were also recorded as secondary outcomes to evaluate the success rate of the prosthetic treatment. The aesthetic parameters were also evaluated according to the FDI criteria. The Kaplan-Meier method and Cox regression with 95% confidence interval were applied for the statistical analysis. RESULTS: Regarding the primary outcome, the metal-ceramic crowns cemented with self-adhesive resin cement presented a high survival rate (91.5%), with 8 crown debondings and 3 tooth losses (1 due to caries and 2 due to periodontal disease) occurring after the evaluation period. For secondary outcomes, 9 root fractures and 4 post debondings occurred, generating a success rate of 72%. All crowns had a score 1 on the FDI criteria, indicating that they were clinically excellent or very good regarding the aesthetic parameters. CONCLUSION: The metal-ceramic crowns luted with a self-adhesive resin cement presented a survival rate of 91.5% after an average of 62 months of follow-up. Furthermore, the restorations remained aesthetically satisfactory over time, without changes that would indicate prosthetic retreatment. A success rate of 72% was obtained considering the secondary outcome, mainly related to intraradicular retainer failures (root fractures or post debonding). CLINICAL SIGNIFICANCE: The self-adhesive resin cement is clinically indicated for cementation of metal-ceramic crowns onto resin composite prosthetic cores.

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Calcium ion regulation plays a crucial role in maintaining neuronal functions such as neurotransmitter release and synaptic plasticity. Copper (Cu2+ ) coordination to amyloid-ß (Aß) has accelerated Aß1-42 aggregation that can trigger calcium dysregulation by enhancing the influx of calcium ions by extensive perturbing integrity of the membranes. Aß1-42 aggregation, calcium dysregulation, and membrane damage are Alzheimer disease (AD) implications. To gain a detail of calcium ions' role in the full-length Aß1-42 and Aß1-42 -Cu2+ monomers contact, the cellular membrane before their aggregation to elucidate the neurotoxicity mechanism, we carried out 2.5 µs extensive molecular dynamics simulation (MD) to rigorous explorations of the intriguing feature of the Aß1-42 and Aß1-42 -Cu2+ interaction with the dimyristoylphosphatidylcholine (DMPC) bilayer in the presence of calcium ions. The outcome of the results compared to the same simulations without calcium ions. We surprisingly noted robust binding energies between the Aß1-42 and membrane observed in simulations containing without calcium ions and is two and a half fold lesser in the simulation with calcium ions. Therefore, in the case of the absence of calcium ions, N-terminal residues of Aß1-42 deeply penetrate from the surface to the center of the bilayer; in contrast to calcium ions presence, the N- and C-terminal residues are involved only in surface contacts through binding phosphate moieties. On the other hand, Aß1-42 -Cu2+ actively participated in surface bilayer contacts in the absence of calcium ions. These contacts are prevented by forming a calcium bridge between Aß1-42 -Cu2+ and the DMPC bilayer in the case of calcium ions presence. In a nutshell, Calcium ions do not allow Aß1-42 penetration into the membranes nor contact of Aß1-42 -Cu2+ with the membranes. These pieces of information imply that the calcium ions mediate the membrane perturbation via the monomer interactions but do not damage the membrane; they agree with the western blot experimental results of a higher concentration of calcium ions inhibit the membrane pore formation by Aß peptides.

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Experimental realizations of two-dimensional materials are hardly free of structural defects such as e.g. vacancies, which, in turn, modify drastically its pristine physical defect-free properties. In this work, we explore effects due to point defect clustering on the electronic and transport properties of bilayer graphene nanoribbons, for AA and AB stacking and zigzag and armchair boundaries, by means of the tight-binding approach and scattering matrix formalism. Evident vacancy concentration signatures exhibiting a maximum amplitude and an universality regardless of the system size, stacking and boundary types, in the density of states around the zero-energy level are observed. Our results are explained via the coalescence analysis of the strong sizeable vacancy clustering effect in the system and the breaking of the inversion symmetry at high vacancy densities, demonstrating a similar density of states for two equivalent degrees of concentration disorder, below and above the maximum value.

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Gramicidin (Gr) nanoparticles (NPs) and poly (diallyl dimethyl ammonium) chloride (PDDA) water dispersions were characterized and evaluated against Gram-positive and Gram-negative bacteria and fungus. Dynamic light scattering for sizing, zeta potential analysis, polydispersity, and colloidal stability over time characterized Gr NPs/PDDA dispersions, and plating and colony-forming units counting determined their microbicidal activity. Cell viabilities of Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans in the presence of the combinations were reduced by 6, 7, and 7 logs, respectively, at 10 µM Gr/10 µg·mL-1 PDDA, 0.5 µM Gr/0. 5µg·mL-1 PDDA, and 0.5 µM Gr/0.5 µg·mL-1 PDDA, respectively. In comparison to individual Gr doses, the combinations reduced doses by half (S. aureus) and a quarter (C. albicans); in comparison to individual PDDA doses, the combinations reduced doses by 6 times (P. aeruginosa) and 10 times (C. albicans). Gr in supported or free cationic lipid bilayers reduced Gr activity against S. aureus due to reduced Gr access to the pathogen. Facile Gr NPs/PDDA disassembly favored access of each agent to the pathogen: PDDA suctioned the pathogen cell wall facilitating Gr insertion in the pathogen cell membrane. Gr NPs/PDDA differential cytotoxicity suggested the possibility of novel systemic uses for the combination.

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Electroporation is a cell-level phenomenon caused by an ionic imbalance in the membrane, being of great relevance in various fields of knowledge. A dependence of the pore formation kinetics on the environmental conditions (temperature and pressure) of the cell membrane has already been reported, but further clarification regarding how these variables affect the pore formation/resealing dynamics and the transport of molecules through the membrane is still lacking. The objective of the present study was to investigate the temperature (288-348 K) and pressure (1-5000 atm) effects on the electroporation kinetics using coarse-grained molecular dynamics simulations. Results shown that the time for pore formation and resealing increased with pressure and decreased with temperature, whereas the maximum pore radius increased with temperature and decreased with pressure. This behavior influenced the ion migration through the bilayer, and the higher ionic mobility was obtained in the 288 K/1000 atm simulations, i.e., a combination of low temperature and (not excessively) high pressure. These results were used to discuss some experimental observations regarding the extraction of intracellular compounds applying this technique. This study contributes to a better understanding of electroporation under different thermodynamic conditions and to an optimal selection of processing parameters in practical applications which exploit this phenomenon.

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Liposomes are artificial models of cellular membranes that are used as delivery systems for genes, drugs and protein antigens. We have previously used them to study the antigenic properties of their phospholipids. Here, we used them to induce the production of IgG anti-non-bilayer phospholipid arrangements (NPAs) antibodies in mice; these antibodies cause cell lysis and trigger a lupus-like disease in mice. We studied the mechanisms that lead to the production of these antibodies, and provide evidence that NK1.1+, CD4+ T cells respond to NPA-bearing liposomes and deliver the help required for specific B cell activation and antibody class-switching to IgG. We found increased numbers of IL-4-producing NK1.1+, CD4+ T cells in the secondary lymphoid organs of mice administered with NPAs, and these cells also expressed CD40L, which is required for B cell activation. Additionally, we isolated and purified NK1.1+, CD4+ T cells from spleens and determined that they over-expressed 40 genes, which are key players in inflammatory processes and B cell stimulation and have TRAF6 and UNC39B1 as key nodes in their network. These results show that liposomes are membrane models that can be used to analyze the immunogenicity of lipids.

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Precise prediction of mechanical behavior of thin films at the nanoscale requires techniques that consider size effects and fabrication-related issues. Here, we propose a test methodology to estimate the Young's modulus of nanometer-thick films using micromachined bilayer cantilevers. The bilayer cantilevers which comprise a well-known reference layer and a tested film deflect due to the relief of the residual stresses generated during the fabrication process. The mechanical relationship between the measured residual stresses and the corresponding deflections was used to characterize the tested film. Residual stresses and deflections were related using analytical and finite element models that consider intrinsic stress gradients and the use of adherence layers. The proposed methodology was applied to low pressure chemical vapor deposited silicon nitride tested films with thicknesses ranging from 46 nm to 288 nm. The estimated Young's modulus values varying between 213.9 GPa and 288.3 GPa were consistent with nanoindentation and alternative residual stress-driven techniques. In addition, the dependence of the results on the thickness and the intrinsic stress gradient of the materials was confirmed. The proposed methodology is simple and can be used to characterize diverse materials deposited under different fabrication conditions.

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Spherical or discoidal lipid polymer nanostructures bearing cationic charges successfully adsorb a variety of oppositely charged antigens (Ag) such as proteins, peptides, nucleic acids, or oligonucleotides. This report provides instructions for the preparation and physical characterization of four different cationic nanostructures able to combine and deliver antigens to the immune system: (1) dioctadecyl dimethylammonium bromide (DODAB) bilayer fragments (DODAB BF); (2) polystyrene sulfate (PSS) nanoparticles (NPs) covered with one cationic dioctadecyl dimethylammonium bromide bilayer (DODAB) named (PSS/DODAB); (3) cationic NPs of biocompatible polymer poly(methyl methacrylate) (PMMA) prepared by emulsion polymerization of the methyl methacrylate (MMA) monomer in the presence of DODAB BF (PMMA/DODAB NPs); (4) antigen NPs (NPs) where the cationic polymer poly(diallyl dimethyl ammonium chloride) (PDDA) directly combined at nontoxic and low dose with the antigen (Ag); when the oppositely charged model antigen is ovalbumin (OVA), NPs are named PDDA/OVA. These nanostructures provide adequate microenvironments for carrying and delivering antigens to the antigen-presenting cells of the immune system.

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Host defense peptides are found primarily as natural antimicrobial agents among all lifeforms. These peptides and their synthetic derivatives have been extensively studied for their potential use as therapeutic agents. The most accepted mechanism of action of these peptides is related to a nonspecific mechanism associated with their interaction with the negatively charged groups present in membranes, inducing bilayer destabilization and cell death through several routes. Among the most recently reported peptides, LTX-315 has emerged as an important oncolytic peptide that is currently in several clinical trials against different cancer types. However, there is a lack of biophysical studies regarding LTX-315 and its interaction with membranes. This research focuses primarily on the understanding of the molecular bases of LTX-315's interaction with eukaryotic lipids, based on two artificial systems representative of non-tumoral and tumoral membranes. Additionally, the interaction with individual lipids was studied by differential scanning calorimetry and Fourier-transformed infrared spectroscopy. The results showed a strong interaction of LTX-315 with the negatively charged phosphatidylserine. The results are important for understanding and facilitating the design and development of improved peptides with anticancer activity.

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Oxidative stress plays an essential role in the regulation of vital processes in living organisms. Reactive oxygen species can react chemically with the constituents of the cells leading to irreversible damage. The first structure of the cell in contact with the environment that surrounds it is the membrane, which protects it and allows the exchange of substances. Some signals manifest when the components of a bilayer are undergoing oxidation, like an increase in the lipid area, decrease in the thickness of the bilayer, and exchange of the oxidized groups toward the bilayer surface. In this investigation, a molecular dynamics simulation was done on a set of Dioleoylphosphatidylcholine membranes with different percentage of oxidized lipids, in order to observe the effect of the oxidation degree on the membrane structure. It was found that, as higher the concentration of oxidized lipids is, the larger the damage of the membrane. This is reflected in the increase in the lipid area and the decrease in the thickness and membrane packing. Also, it was observed that hydrophobicity inside the membrane decreases as the oxidation percentage increases.Communicated by Ramaswamy H. Sarma.

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Tissue engineering has focused on the development of biomaterials that emulate the native extracellular matrix. Therefore, the purpose of this research was oriented to the development of nanofibrillar bilayer membranes composed of polycaprolactone with low and medium molecular weight chitosan, evaluating their physicochemical and biological properties. Two-bilayer membranes were developed by an electrospinning technique considering the effect of chitosan molecular weight and parameter changes in the technique. Subsequently, the membranes were evaluated by scanning electron microscopy, Fourier transform spectroscopy, stress tests, permeability, contact angle, hemolysis evaluation, and an MTT test. From the results, it was found that changes in the electrospinning parameters and the molecular weight of chitosan influence the formation, fiber orientation, and nanoarchitecture of the membranes. Likewise, it was evidenced that a higher molecular weight of chitosan in the bilayer membranes increases the stiffness and favors polar anchor points. This increased Young's modulus, wettability, and permeability, which, in turn, influenced the reduction in the percentage of cell viability and hemolysis. It is concluded that the development of biomimetic bilayer nanofibrillar membranes modulate the physicochemical properties and improve the hemolytic behavior so they can be used as a hemocompatible biomaterial.

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OBJECTIVE: To understand the stress development in porcelain-veneered zirconia (PVZ) and porcelain-veneered lithium disilicate (PVLD) crowns with different veneer/core thickness ratios and cooling rates. To provide design guidelines for better performing bilayer restorations with the aid of Viscoelastic Finite Element Method (VFEM). METHODS: The VFEM was validated by comparing the predicted residual stresses with experimental measurements. Then, the model was used to predict transient and residual stresses in the two bilayer systems. Models with two different veneer/core thickness ratios were prepared (2:1 and 1:1) and two cooling protocols were simulated (Fast: ∼300 °C/min, Slow: ∼30 °C/min) using the heat transfer module, followed by stress analysis in ABAQUS. The physical properties of zirconia, lithium disilicate, and the porcelains used for the simulations were determined as a function of temperature. RESULTS: PVLD showed lower residual stresses than PVZ. The maximum tensile stresses in PVZ were observed in the cusp area, whereas those in PVLD were located in the central fossa. The 1:1 thickness ratio decreased stresses in both layers of PVZ. Slow cooling slightly decreased residual stresses in both systems. However, the cooling rate effect was more evident in transient stresses. SIGNIFICANCE: Slow cooling is preferable for both systems. A thinner porcelain layer over zirconia lowers stresses throughout the restoration. The different stress distributions between PVZ and PVLD may affect their failure modes. Smaller mismatches in modulus, CTE, and specific heat between the constituents, and the use of low Tg porcelains can effectively reduce the deleterious transient and residual tensile stresses in bilayer restorations.

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