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The versatility of metal complexes of corroles has raised interest in the use of these molecules as elements of chemical sensors. The tuning of the macrocycle properties via synthetic modification of the different components of the corrole ring, such as functional groups, the molecular skeleton, and coordinated metal, allows for the creation of a vast library of corrole-based sensors. However, the scarce conductivity of most of the aggregates of corroles limits the development of simple conductometric sensors and requires the use of optical or mass transducers that are rather more cumbersome and less prone to be integrated into microelectronics systems. To compensate for the scarce conductivity, corroles are often used to functionalize the surface of conductive materials such as graphene oxide, carbon nanotubes, or conductive polymers. Alternatively, they can be incorporated into heterojunction devices where they are interfaced with a conductive material such as a phthalocyanine. Herewith, we introduce two heterostructure sensors combining lutetium bisphthalocyanine (LuPc2) with either 5,10,15-tris(pentafluorophenyl) corrolato Cu (1) or 5,10,15-tris(4-methoxyphenyl)corrolato Cu (2). The optical spectra show that after deposition, corroles maintain their original structure. The conductivity of the devices reveals an energy barrier for interfacial charge transport for 1/LuPc2, which is a heterojunction device. On the contrary, only ohmic contacts are observed in the 2/LuPc2 device. These different electrical properties, which result from the different electron-withdrawing or -donating substituents on corrole rings, are also manifested by the opposite response with respect to ammonia (NH3), with 1/LuPc2 behaving as an n-type conductor and 2/LuPC2 behaving as a p-type conductor. Both devices are capable of detecting NH3 down to 10 ppm at room temperature. Furthermore, the sensors show high sensitivity with respect to relative humidity (RH) but with a reversible and fast response in the range of 30-60% RH.
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The mucosal pellicle, also called salivary pellicle, is a thin biological layer made of salivary and epithelial constituents, lining oral mucosae. It contributes to their protection against microbiological, chemical, or mechanical insults. Pellicle formation depends on the cells' surface properties, and in turn the pellicle deeply modifies such properties. It has been reported that the expression of the transmembrane mucin MUC1 in oral epithelial cells improves the formation of the mucosal pellicle. Here, we describe an approach combining classical and functionalized tip atomic force microscopy and scanning microwave microscopy to characterize how MUC1 induces changes in buccal cells' morphology, hydrophobicity, and electric properties to elucidate the physicochemical mechanisms involved in the enhancement of the anchoring of salivary proteins. We show that MUC1 expression did not modify drastically the morphology of the epithelial cells' surface. MUC1 expression, however, resulted in the presence of more hydrophobic and more charged areas at the cell surface. The presence of salivary proteins decreased the highest attractive and repulsive forces recorded between the cell surface and a functionalized hydrophobic atomic force microscopy (AFM) tip, suggesting that the most hydrophobic and charged areas participate in the binding of salivary proteins. The cells' dielectric properties were altered by both MUC1 expression and the presence of a mucosal pellicle. We finally show that in the absence of MUC1, the pellicle appeared as a distinct layer poorly interacting with the cells' surface. This integrative AFM/scanning microwave microscopy approach may usefully describe the surface properties of various cell types, with relevance to the bioadhesion or biomimetics fields.
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Boca/citología , Nanotecnología , Saliva/metabolismo , Impedancia Eléctrica , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Propiedades de SuperficieRESUMEN
Studies on human norovirus are severely hampered by the absence of a cell culture system until the discovery of murine norovirus (MNV). The cell membrane domains called lipid rafts have been defined as a port of entry for viruses. This study is conducted to investigate murine norovirus binding on the mouse leukemic monocyte macrophage cell line. Lipid raft related structures are extracted from cells by detergent treatment resulting detergent-resistant membrane (DRMs) domains. The real-time polymerase chain reaction technique is performed to detect the viral genome, thereby the MNV binding on the DRMs. The interactions between MNV and DRMs are investigated by high-speed atomic force microscopy (HS-AFM) combined with surface-enhanced Raman spectroscopy (SERS). The inoculation of the virus onto cells results in the aggregations of detergent-resistant membrane domains significantly. The characteristic Raman band of MNV is found in inoculated samples. To be sure that these results are originated from specific interactions between DRM and MNV, methyl-ß-cyclo-dextrin (MßCD) is applied to disrupt lipid rafts. The MNV binding on DRMs is precluded by the MßCD treatment. The cholesterols chains are defined as a key factor in the interactions between norovirus and DRMs. The authors conclude that the MNV binding involves the presence of DRMs and cholesterol dependent.
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Infecciones por Caliciviridae/metabolismo , Microdominios de Membrana/metabolismo , Microscopía de Fuerza Atómica/métodos , Norovirus/fisiología , Espectrometría Raman/métodos , Animales , Microdominios de Membrana/efectos de los fármacos , Ratones , Células RAW 264.7 , Reacción en Cadena en Tiempo Real de la Polimerasa , beta-Ciclodextrinas/farmacologíaRESUMEN
Estrothiazine (ESTZ) is a weak estrogen sharing structural similarities with coumestrol. ESTZ failed to compete with [3H]17ß-estradiol ([3H]17ß-E2) for binding to the estrogen receptor α (ERα), questioning its ability to interact with the receptor. However, detection by atomic force spectroscopy (AFS) of an ESTZ-induced ERα dimerization has eliminated any remaining doubts. The effect of the compound on the proliferation of ERα-positive and negative breast cancer cells confirmed the requirement of the receptor. The efficiency of ESTZ in MCF-7 cells was weak without any potency to modify the proliferation profile of estradiol and coumestrol. Growth enhancement was associated with a proteasomal degradation of ERα without substantial recruitment of LxxLL coactivators. This may be related to an unusual delay between the acquisition by the receptor of an ERE-binding capacity and the subsequent estrogen-dependent transcription. A complementary ability to enhance TPA-induced AP-1 transcription was observed, even at concentrations insufficient to activate the ERα, suggesting a partly independent mechanism. ESTZ also rapidly and transiently activated ERK1/2 likely through membrane estrogenic pathways provoking a reorganization of the actin network. Finally, the systematic absence of biological responses with an ESTZ derivative unable to induce ERα dimerization stresses the importance of this step in the action of the compound, as reported for conventional estrogens. In view of the existence of many other ERα modulators (endocrine disruptors such as, for example, pesticides, environmental contaminants or phytoestrogens) with extremely weak or similar apparent lack of binding ability, our work may appear as pilot investigation for assessing their mechanism of action.
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Neoplasias de la Mama/tratamiento farmacológico , Receptor alfa de Estrógeno/metabolismo , Tiazinas/administración & dosificación , Transcripción Genética , Sitios de Unión , Neoplasias de la Mama/genética , Dimerización , Estradiol/metabolismo , Receptor alfa de Estrógeno/genética , Femenino , Humanos , Células MCF-7 , Fitoestrógenos , Unión Proteica/genética , Espectrofotometría Atómica , Tiazinas/química , Factor de Transcripción AP-1/genética , Factor de Transcripción AP-1/metabolismoRESUMEN
Metallic nanoparticles are considered as active supports in the development of specific chemical or biological biosensors. Well-organized nanoparticles can be prepared either through expensive (e.g., electron beam lithography) or inexpensive (e.g., thermal synthesis) approaches where different shapes of nanoparticles are easily obtained over large solid surfaces. Herein, the authors propose a low-cost thermal synthesis of active plasmonic nanostructures on thin gold layers modified glass supports after 1 h holding on a hot plate (~350 °C). The resulted annealed nanoparticles proved a good reproducibility of localized surface plasmon resonance (LSPR) and surface enhanced Raman spectroscopy (SERS) optical responses and where used for the detection of low concentrations of two model (bio)chemical molecules, namely the human cytochrome b5 (Cyt-b5) and trans-1,2-bis(4-pyridyl)ethylene (BPE).
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Nanoestructuras , Oro , Reproducibilidad de los Resultados , Espectrometría Raman , Resonancia por Plasmón de SuperficieRESUMEN
We study the aggregation of calcium silicate hydrate nanoplatelets on a surface by means of Monte Carlo and molecular dynamics simulations at thermodynamic equilibrium. Calcium silicate hydrate (C-S-H) is the main component formed in cement and is responsible for the strength of the material. The hydrate is formed in early cement paste and grows to form platelets on the nanoscale, which aggregate either on dissolving cement particles or on auxiliary particles. The general result is that the experimentally observed variations in these dynamic processes generically called growth can be rationalized from interaction free energies, that is, from pure thermodynamic arguments. We further show that the surface charge density of the particles determines the aggregate structures formed by C-S-H and thus their growth modes.
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Scientists are currently truly committed to enhance the specificity of chemotherapeutics that target DNA. To this end, sequence-specific drugs have progressively given way to structure-specific therapeutics. However, while numerous strategies have been implemented to design high-affinity candidates, strategies devoted to the design of high-selectivity ligands are still rare. Here we report on such an approach via the study of an amphiphilic compound, TEGPy, that self-assembles at a liquid/solid interface to provide nanosized objects that are stable in water. The resulting aggregates, identified through atomic force microscopy measurements, were found to disassemble upon interaction with DNA in a structure-specific manner (quadruplex- versus duplex-DNA). Our results provide a fertile ground for devising new strategies aiming at concomitantly enhancing DNA structural specificity and the water-solubility of aggregation-prone ligands.
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ADN/química , Porfirinas/química , Tensoactivos/química , G-Cuádruplex , Ligandos , Microscopía de Fuerza Atómica , Modelos Moleculares , Nanoestructuras/química , Nanoestructuras/ultraestructura , Conformación de Ácido NucleicoRESUMEN
Modulation of interfacial conductivity in organic heterostructures is a highly promising strategy to improve the performance of electronic devices. In this endeavor, the present work reports the fabrication of a bilayer heterojunction device, combining octafluoro copper phthalocyanine (CuF8Pc) and lutetium bis-phthalocyanine (LuPc2) and tunes the charge transport at the Cu(F8Pc)-(LuPc2) interface by aryl electrografting on the device electrode to improve the device NH3-sensing properties. Dimethoxybenzene (DMB) and tetrafluoro benzene (TFB) electrografted by an aryldiazonium electroreduction method form a few-nanometer-thick organic film on ITO. The conductivity of the heterojunction devices formed by coating a Cu(F8Pc)/LuPc2 bilayer over the aryl-grafted electrode strongly varies according to the electronic effects of the substituents in the aryl. Accordingly, DMB increases while TFB decreases the mobile charges accumulation at the Cu(F8Pc)-(LuPc2) interface. This is explained by the perfect alignment of the frontier molecular orbitals of DMB and Cu(F8Pc), facilitating charge injection into the Cu(F8Pc) layer. On the contrary, TFB behaves like a strong acceptor and reduces the mobile charges accumulation at the Cu(F8Pc)-(LuPc2) interface. Such interfacial conductivity variation influences the device NH3-sensing properties, which increase because of DMB grafting and decrease in the presence of TFB. DMB-based heterojunction devices contain four times higher active sites for NH3 adsorption and could detect NH3 down to 1 ppm with limited interference from humidity, making them suitable for real environment NH3 detection.
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Conventional techniques that measure the concentration of light elements in metallic materials lack high-resolution performance due to their intrinsic limitation of sensitivity. In that context, scanning microwave microscopy has the potential to significantly enhance the quantification of element distribution due to its ability to perform a tomographic investigation of the sample. Scanning microwave microscopy associates the local electromagnetic measurement and the nanoscale resolution of an atomic force microscope. This technique allows the simultaneous characterization of oxygen concentration as well as local mechanical properties by microwave phase shift and amplitude signal, respectively. The technique was calibrated by comparison with nuclear reaction analysis and nanoindentation measurement. We demonstrated the reliability of the scanning microwave technique by studying thin oxygen-enriched layers on a Ti-6Al-4V alloy. This innovative approach opens novel possibilities for the indirect quantification of light chemical element diffusion in metallic materials. This technique is applicable to the control and optimization of industrial processes.
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The mucosal pellicle (MP) is a biological film protecting the oral mucosa. It is composed of bounded salivary proteins and transmembrane mucin MUC1 expressed by oral epithelial cells. Previous research indicates that MUC1 expression enhances the binding of the main salivary protein forming the MP, MUC5B. This study investigated the influence of MUC1 structure on MP formation. A TR146 cell line, which does not express MUC1 natively, was stably transfected with genes coding for three MUC1 isoforms differing in the structure of the two main extracellular domains: the VNTR domain, exhibiting a variable number of tandem repeats, and the SEA domain, maintaining the two bound subunits of MUC1. Semi-quantification of MUC1 using dot blot chemiluminescence showed comparable expression levels in all transfected cell lines. Semi-quantification of MUC5B by immunostaining after incubation with saliva revealed that MUC1 expression significantly increased MUC5B adsorption. Neither the VNTR domain nor the SEA domain was influenced MUC5B anchoring, suggesting the key role of the MUC1 N-terminal domain. AFM-IR nanospectroscopy revealed discernible shifts indicative of changes in the chemical properties at the cell surface due to the expression of the MUC1 isoform. Furthermore, the observed chemical shifts suggest the involvement of hydrophobic effects in the interaction between MUC1 and salivary proteins.
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Extracellular vesicles (EVs) are membrane-derived, tiny vesicles produced by all cells that range from 50 to several hundreds of nanometers in diameter and are used as a means of intercellular communication. They are emerging as promising diagnostic and therapeutic tools for a variety of diseases. There are two main biogenesis processes used by cells to produce EVs with differences in size, composition, and content. Due to their high complexity in size, composition, and cell origin, their characterization requires a combination of analytical techniques. This project involves the development of a new generation of multiparametric analytical platforms with increased throughput for the characterization of subpopulations of EVs. To achieve this goal, the work starts from the nanobioanalytical platform (NBA) established by the group, which allows an original investigation of EVs based on a combination of multiplexed biosensing methods with metrological and morphomechanical analyses by atomic force microscopy (AFM) of vesicular targets trapped on a microarray biochip. The objective was to complete this EV investigation with a phenotypic and molecular analysis by Raman spectroscopy. These developments enable the proposal of a multimodal and easy-to-use analytical solution for the discrimination of EV subsets in biological fluids with clinical potential.
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Técnicas Biosensibles , Vesículas Extracelulares , Resonancia por Plasmón de Superficie , Vesículas Extracelulares/química , Microscopía de Fuerza Atómica/métodos , Comunicación CelularRESUMEN
With the growing interest in environmentally friendly and personalized medicines, new concept for combining three-dimensional printing (3DP) with natural-based biomaterials derived from agro-food wastes has emerged. This approach provides sustainable solutions for agricultural waste management and potential for developing of novel pharmaceutical products with tunable characteristics. This work demonstrated the feasibility of fabricating personalized theophylline films with four different structures (Full, Grid, Star, and Hilbert) using syringe extrusion 3DP and carboxymethyl cellulose (CMC) derived from durian rind wastes. Our findings suggested that all the CMC-based inks with shear thinning properties capable of being extruded smoothly through a small nozzle could potentially be used to fabricate the films with various complex printing patterns and high structural fidelity. The results also demonstrated that the film characteristics and release profiles could be easily modified by simply changing the slicing parameters (e.g., infill density and printing pattern). Amongst all formulations, Grid film, which was 3D-printed with 40 % infill and a grid pattern, demonstrated a highly porous structure with high total pore volume. The voids between printing layers in Grid film increased theophylline release (up to 90 % in 45 min) through improved wetting and water penetration. All findings in this study provide significant insight into how to modify film characteristics simply by digitally changing the printing pattern in slicer software without creating a new CAD model. This approach could help to simplify the 3DP process so that non-specialist users can easily implement it in community pharmacies or hospital on demand.
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Bombacaceae , Carboximetilcelulosa de Sodio , Tinta , Teofilina , Impresión TridimensionalRESUMEN
Spatial localizing of skeletal proteins in biogenic minerals remains a challenge in biomineralization research. To address this goal, we developed a novel in situ mapping technique based on molecular recognition measurements via atomic force microscopy (AFM), which requires three steps: (1) the development and purification of a polyclonal antibody elicited against the target protein, (2) its covalent coupling to a silicon nitride AFM tip ('functionalization'), and (3) scanning of an appropriately prepared biomineral surface. We applied this approach to a soluble shell protein - accripin11 - recently identified as a major component of the calcitic prisms of the fan mussel Pinna nobilis [1]. Multiple tests reveal that accripin11 is evenly distributed at the surface of the prisms and also present in the organic sheaths surrounding the calcitic prisms, indicating that this protein is both intra- and inter-crystalline. We observed that the adhesion force in transverse sections is about twice higher than in longitudinal sections, suggesting that accripin11 may exhibit preferred orientation in the biomineral. To our knowledge, this is the first time that a protein is localized by molecular recognition atomic force microscopy with antibody-functionalized tips in a biogenic mineral. The 'pros' and 'cons' of this methodology are discussed in comparison with more 'classical' approaches like immunogold. This technique, which leaves the surface to analyze clean, might prove useful for clinical tests on non-pathological (bone, teeth) or pathological (kidney stone) biomineralizations. Studies using implants with protein-doped calcium phosphate coating can also benefit from this technology. STATEMENT OF SIGNIFICANCE: Our paper deals with an unconventional technical approach for localizing proteins that are occluded in biominerals. This technique relies on the use of molecular recognition atomic force microscopy with antibody-functionalized tips. Although such approach has been employed in other system, this is the very first time that it is developed for biominerals. In comparison to more classical approaches (such as immunogold), AFM microscopy with antibody-functionalized tips allows higher magnification and keeps the scanned surface clean for other biophysical characterizations. Our method has a general scope as it can be applied in human health, for non-pathological (bone, teeth) and pathological (kidney stone) biomineralizations as well as for bone implants coated with protein-doped calcium phosphate.
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Bivalvos , Cálculos Renales , Animales , Humanos , Microscopía de Fuerza Atómica/métodos , Proteínas/química , Anticuerpos , Carbonato de Calcio/metabolismo , Fosfatos de CalcioRESUMEN
BACKGROUND: Quantitative modeling of the self-assembly of DNA tiles leading either to defined end-products or distribution of biopolymers is of practical importance for biotechnology and synthetic biology. METHODS: The combinatorial process describing tile assembly was implemented into a generic algorithm allowing quantitative description of the population of significant species accumulating during the reaction course. Experimental formation and characterization by optical and electrophoresis approaches of copolymers resulting from the self-assembly of a limited number of half-complementary tiles were used to define and validate generic rules allowing definition of model parameters. RESULTS: Factors controlling the structure and the dynamic of the oligomer population were evidenced for assemblies leading or not to defined end-products. Primary parameters were experimentally determined using rapid mixing experiments. Adjustment of simulations to experimental profiles allowed definition of generic rules for calculation of secondary parameters that take into account macro- and microenvironment of individual hybridization steps. In the case of copolymers, accurate simulation of experimental profiles was achieved for formation of linear assemblies. CONCLUSIONS: Overall length of species and structure of the DNA regions flanking the hybridization sites are critical parameters for which calculation rules were defined. The computational approach quantitatively predicted the parameters affecting time-course and distribution of accumulating products for different experimental designs. GENERAL SIGNIFICANCE: The computational and parameter evaluation procedures designed for the assembly of DNA tiles into large 1D-structures are more generally applicable for the construction of non-DNA polymers by extremities-specific recognition of molecular blocks.
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ADN/química , Modelos Genéticos , Secuencia de Bases , Técnicas Químicas Combinatorias , Simulación por Computador , ADN/genética , Diseño de Fármacos , Cinética , Datos de Secuencia Molecular , Reproducibilidad de los ResultadosRESUMEN
Significant efforts have been done in last two decades to develop nanoscale spectroscopy techniques owning to their great potential for single-molecule structural detection and in addition, to resolve open questions in heterogeneous biological systems, such as protein-DNA complexes. Applying IR-AFM technique has become a powerful leverage for obtaining simultaneous absorption spectra with a nanoscale spatial resolution for studied proteins, however the AFM-IR investigation of DNA molecules on surface, as a benchmark for a nucleoprotein complexes nanocharacterization, has remained elusive. Herein, we demonstrate methodological approach for acquisition of AFM-IR mapping modalities with corresponding absorption spectra based on two different DNA deposition protocols on spermidine and Ni2+ pretreated mica surface. The nanoscale IR absorbance of distinctly formed DNA morphologies on mica are demonstrated through series of AFM-IR absorption maps with corresponding IR spectrum. Our results thus demonstrate the sensitivity of AFM-IR nanospectroscopy for a nucleic acid research with an open potential to be employed in further investigation of nucleoprotein complexes.
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Silicatos de Aluminio , ADN , Microscopía de Fuerza Atómica/métodos , Silicatos de Aluminio/química , Nucleoproteínas , Espectrofotometría Infrarroja/métodosRESUMEN
This work aims to synthesize polygalacturonate-based magnetic iron oxide nanoparticles (INP-polyGalA). The synthesis consists of the diffusion of both Fe2+ and Fe3+ at a molar ratio of 1:2 through polyGalA solution followed by the addition of an alkaline solution. To form individual nanoparticle materials, the polyGalA concentration needs to be below its overlapping concentration (C*). The synthesized materials (INP-polyGalA) contain about 45% of organic compound (polyGalA), and they have an average particle size ranging from 10 to 50 nm as estimated by several techniques (DLS, TEM and AFM) and their surfaces are negatively charged in pH range 2 to 7. The synthesized NPs showed magnetic characteristics, thanks to the formation of magnetite (Fe3O4) as confirmed by X-ray diffractions (XRD). Moreover, AFM combined with Infra-red mapping allowed us to conclude that polyGalA is located in the core of the nanoparticles but also on their surfaces. More specially, both carboxylate (COO-) and carboxylic (COOH) groups of polyGalA are observed on the NPs surfaces. The presence of such functional groups allowed the synthesized material to (i) bind through the electrostatic interactions methylene blue (MB) which may have a great potential for r pollution control or (ii) to form hydrogel beads (ionotropic gelation) by using calcium as a crosslinking agent which can be used to encapsulate active molecules and target their release by using an external stimulus (magnetic field).
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Nanopartículas de Magnetita , Nanopartículas , Calcio , Óxido Ferrosoférrico , Hidrogeles , Nanopartículas Magnéticas de Óxido de Hierro , Nanopartículas de Magnetita/química , Azul de MetilenoRESUMEN
Elucidating origin, composition, size, and lifetime of microdomains in biological membranes remains a major issue for the understanding of cell biology. For lipid domains, the lack of a direct access to the behaviour of samples at the mesoscopic scale has constituted for long a major obstacle to their characterization, even in simple model systems made of immiscible binary mixtures. By its capacity to image soft surfaces with a resolution that extends from the molecular to the microscopic level, in air as well as under liquid, atomic force microscopy (AFM) has filled this gap and has become an inescapable tool in the study of the surface topography of model membrane domains, the first essential step for the understanding of biomembranes organization. In this review we mainly focus on the type of information on lipid microdomains in model systems that only AFM can provide. We will also examine how AFM can contribute to understand data acquired by a variety of other techniques and present recent developments which might open new avenues in model and biomembrane AFM applications.
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Membrana Dobles de Lípidos/química , Lípidos de la Membrana/química , Microdominios de Membrana/química , Proteínas de la Membrana/química , Animales , Colesterol/química , Humanos , Microscopía de Fuerza Atómica , Esfingolípidos/químicaRESUMEN
Synthetic proteo-nucleic structures (PDNAs) encompassing a single-stranded DNA sequence covalently attached to a redox protein domain able to interact with surface or matrix were designed and characterized. They constitute versatile building blocks alternative to regular DNA for creating scaffolds with optical, electrical, or catalytic properties. PDNAs self-assemble in the presence of complementary oligonucleotides, to form a network of protein domains linked by double-stranded DNA segments. Electrophoretic and hydrodynamic behaviors of PDNAs and corresponding DNA were compared under electrophoresis and gel filtration conditions. Hybridization rates between small and large assemblies were characterized by rapid-mixing experiments. Results showed that the protein part significantly contributes to hydrodynamic behaviors of structures but marginally affects the conformation and hybridization properties of the nucleic domain. PDNA metal-mediated complexes with nitriloacetate-modified phospholipids can diffuse and interact at the surface of vesicles or supported membranes. Surface plasmon resonance analysis of membrane-PDNA interactions indicated that two protein units are required to allow stable surface association and that surface occupancy constrains assembly sizes. High-speed atomic force microscopy illustrated rapid lateral diffusion of assemblies on mica, revealing transient association between noncomplementary PDNA extremities and frequent trapping by surface defects. Regularly organized protein domains were visualized using a larger DNA framework.
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ADN/química , Proteínas/química , Silicatos de Aluminio , Cromatografía en Gel , Citocromos b/química , Metales/química , Microscopía de Fuerza Atómica , Estructura Molecular , Soluciones , Resonancia por Plasmón de SuperficieRESUMEN
Although relative air humidity (RH) strongly influences microbial survival, its use for fighting surface pathogens in the food industry has been inadequately considered. We asked whether RH control could destroy Listeria monocytogenes EGDe by envelope damage. The impact of dehydration in phosphate-buffered saline (PBS) at 75%, 68%, 43% and 11% RH on the bacterial envelope was investigated using flow cytometry and atomic force microscopy. Changes after rehydration in the protein secondary structure and peptidoglycan were investigated by infrared spectroscopy. Complementary cultivability measurements were performed by running dehydration-rehydration with combinations of NaCl (3-0.01%), distilled water, city water and PBS. The main results show that cell membrane permeability and cell envelope were greatly altered during dehydration in PBS at 68% RH followed by rapid rehydration. This damage led cells to recover only 67% of their initial volume after rehydration. Moreover, the most efficient way to destroy cells was dehydration and rehydration in city water. Our study indicates that rehydration of dried, sullied foods on surfaces may improve current cleaning procedures in the food industry.
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Flavor is one of the main drivers of food consumption and acceptability. It is associated with pleasure feels during eating. Flavor is a multimodal perception corresponding to the functional integration of information from the chemical senses: olfaction, gustation, and nasal and oral somatosensory inputs. As a result, astringency, as a sensation mediated by the trigeminal nerves, influences food flavor. Despite the importance of astringency in food consumer acceptance, the exact chemosensory mechanism of its detection and the nature of the receptors activated remain unknown. Herein, after reviewing the current hypotheses on the molecular origin of astringency, we proposed a ground-breaking hypothesis on the molecular mechanisms underpinning this sensation as a perspective for future research.