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The correct establishment of DNA methylation patterns is vital for mammalian development and is achieved by the de novo DNA methyltransferases DNMT3A and DNMT3B. DNMT3B localises to H3K36me3 at actively transcribing gene bodies via its PWWP domain. It also functions at heterochromatin through an unknown recruitment mechanism. Here, we find that knockout of DNMT3B causes loss of methylation predominantly at H3K9me3-marked heterochromatin and that DNMT3B PWWP domain mutations or deletion result in striking increases of methylation in H3K9me3-marked heterochromatin. Removal of the N-terminal region of DNMT3B affects its ability to methylate H3K9me3-marked regions. This region of DNMT3B directly interacts with HP1α and facilitates the bridging of DNMT3B with H3K9me3-marked nucleosomes in vitro. Our results suggest that DNMT3B is recruited to H3K9me3-marked heterochromatin in a PWWP-independent manner that is facilitated by the protein's N-terminal region through an interaction with a key heterochromatin protein. More generally, we suggest that DNMT3B plays a role in DNA methylation homeostasis at heterochromatin, a process which is disrupted in cancer, aging and Immunodeficiency, Centromeric Instability and Facial Anomalies (ICF) syndrome.
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Metilación de ADN , Cara/anomalías , Heterocromatina , Enfermedades de Inmunodeficiencia Primaria , Animales , ADN (Citosina-5-)-Metiltransferasas/genética , ADN (Citosina-5-)-Metiltransferasas/metabolismo , ADN Metiltransferasa 3A , Mutación , Mamíferos/genética , Mamíferos/metabolismoRESUMEN
This work investigates the effect of dilution on the phase separation process of binary charged polysaccharide-surfactant mixtures formed by two cationic polysaccharides and up to four surfactants of different nature (anionic, zwitterionic, and neutral), as well as the potential impact of dilution-induced phase separation on the formation of conditioning deposits on charged surfaces, mimicking the negative charge and wettability of damaged hair fibers. The results obtained showed that the dilution behavior of model washing formulations (concentrated polysaccharide-surfactant mixtures) cannot be described in terms of a classical complex precipitation framework, as phase separation phenomena occur even when the aggregates are far from the equilibrium phase separation composition. Therefore, dilution-enhanced deposition cannot be predicted in terms of the worsening of colloidal stability due to the charge neutralization phenomena, as common phase separation and, hence, enhanced deposition occurs even for highly charged complexes.
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In recent years, the availability of effective vaccines has become a public health challenge due to the proliferation of different pandemic outbreaks which are a risk for the world population health. Therefore, the manufacturing of new formulations providing a robust immune response against specific diseases is of paramount importance. This can be partially faced by introducing vaccination systems based on nanostructured materials, and in particular, nanoassemblies obtained by the Layer-by-Layer (LbL) method. This has emerged, in recent years, as a very promising alternative for the design and optimization of effective vaccination platforms. In particular, the versatility and modularity of the LbL method provide very powerful tools for fabricating functional materials, opening new avenues on the design of different biomedical tools, including very specific vaccination platforms. Moreover, the possibility to control the shape, size, and chemical composition of the supramolecular nanoassemblies obtained by the LbL method offers new opportunities for manufacturing materials which can be administered following specific routes and present very specific targeting. Thus, it will be possible to increase the patient convenience and the efficacy of the vaccination programs. This review presents a general overview on the state of the art of the fabrication of vaccination platforms based on LbL materials, trying to highlight some important advantages offered by these systems.
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The association of polyelectrolytes and surfactants bearing charges of opposite signs has been for long time considered under an equilibrium framework. However, this is far to provide a true description of the physico-chemical rules of the association process when kinetically arrested nonequilibrium states are formed. This is the result of specific interaction pathways between the polyelectrolyte chains and the surfactant molecules determined by the mixing procedure and the mixture composition. In fact, the specific characteristics of the method used for the mixture can induce local inhomogeneities in the mixture composition which can govern the properties of the obtained supramolecular aggregates, driving to the systems to a situation far from the true equilibrium. This topical review tries to provide to the reader a general perspective of the role of the nonequilibrium aspects in the control of the polyelectrolyte-surfactant association process, and how these impact on the obtained supramolecular nanoassemblies, and their properties.
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Diffusion of biological macromolecules in the cytoplasm is a paradigm of colloidal diffusion in an environment characterized by a strong restriction of the accessible volume. This makes of the understanding of the physical rules governing colloidal diffusion under conditions mimicking the reduction in accessible volume occurring in the cell cytoplasm, a problem of a paramount importance. This work aims to study how the thermal motion of spherical colloidal beads in the inner cavity of giant unilamellar vesicles (GUVs) is modified by strong confinement conditions, and the viscoelastic character of the medium. Using single particle tracking, it is found that both the confinement and the environmental viscoelasticity lead to the emergence of anomalous motion pathways for colloidal microbeads encapsulated in the aqueous inner cavity of GUVs. This anomalous diffusion is strongly dependent on the ratio between the volume of the colloidal particle and that of the GUV under consideration as well as on the viscosity of the particle's liquid environment. Therefore, the results evidence that the reduction of the free volume accessible to colloidal motion pushes the diffusion far from a standard Brownian pathway as a result of the change in the hydrodynamic boundary conditions driving the particle motion.
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HYPOTHESIS: The internal organization of polyelectrolyte layers deposited on colloidal templates plays a very important role for the potential applications of these systems as capsules for drug delivery purposes. EXPERIMENTS: The mutual arrangement of oppositely charged polyelectrolyte layers upon their deposition on positively charged liposomes has been studied by combining up three different scattering techniques and Electronic Spin Resonance, which has provided information about the inter-layer interactions and their effect on the final structure of the capsules. FINDINGS: The sequential deposition of oppositely charged polyelectrolytes on the external leaflet of positively charged liposomes allows modulating the organization of the obtained supramolecular structures, impacting the packing and rigidity of the obtained capsules due to the change of the ionic cross-linking of the multi-layered film as a result of the specific charge of the last deposited layer. The possibility to modulate the properties of the LbL capsules by tuning the characteristics of the last deposited layers offers a very interesting route for the design of materials for encapsulation purposes with their properties controlled almost at will by changing the number of deposited layers and their chemistry.
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The concentration dependence of the surface tension of several binary mixtures of non-electrolytes has been measured at 298.15 K. The mixtures have been chosen since they presented a so-called "W-shape" concentration dependence of the excess constant pressure heat capacity and high values of the concentration-concentration correlation function. This behavior was interpreted in terms of the existence of anomalously high concentration fluctuations that resemble those existing in the proximities of critical points. However, no liquid-liquid phase separation has been found in any of these mixtures over a wide temperature range. In this work, we have extended these studies to the liquid-air interfacial properties. The results show that the concentration dependence of the surface tension shows a plateau and the mixing surface tension presents a "W-shape" behavior. To the best of our knowledge, this is the first time that this behavior is reported. The weak anomalies of the surface tension near a liquid-liquid critical point suggest that the results obtained cannot be considered far-from-critical effects. The usual approach of substituting the activity by the concentration in the Gibbs equation for the relative surface concentration has been found to lead to large errors and the mixtures to have a fuzzy and thick liquid/vapor interface.
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Calor , Tensión SuperficialRESUMEN
The interaction of particles with fluid interfaces is ubiquitous in synthetic and natural work, involving two types of interactions: particle-interface interactions (trapping energy) and interparticle interactions. Therefore, it is urgent to gain a deep understanding of the main forces controlling the trapping of particles at fluid interfaces, and their assembly to generate a broad range of structures characterized by different degrees of order. This Perspective tries to provide an overview of the main contributions to the energetic landscape controlling the assembly of particles at fluid interfaces, which is essential for exploiting this type of interfacial systems as platforms for the fabrication of interface-based soft materials with technological interest.
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Mixtures of polyelectrolytes and surfactants are commonly used in many technological applications where the challenge is to provide well-defined modifications of the surface properties, as is the case of washing formulations in cosmetics. However, if contemporary experimental and theoretical methods can provide insights on their behavior in concentrated formulations, less is known on their behavior under practical use conditions, e.g., under dilution and vectorization of deposits. This makes it difficult to make predictions for specific performance, as, for example, good hair manageability after a shampoo or a comfortable sensorial appreciation after a skin cleanser. This is especially important when considering the formulation of new, more eco-friendly formulations. In this work, a detailed study of the phase separation process induced by dilution is described, as well as the impact on the deposition of conditioning material on negatively charged surfaces. In order to gain a more detailed physical insight, several polyelectrolyte-surfactant pairs, formed by two different polymers and five surfactants that, although non-natural or eco-friendly, can be considered as models of classical formulations, have been studied. The results evidenced that upon dilution the behavior, and hence its deposition onto the surface, cannot be predicted in terms of the behavior of simpler pseudo-binary (mixtures of a polymer and a surfactant) or pseudo-ternary mixtures (two polymers and a surfactant). In many cases, phase separation was observed for concentrations similar to those corresponding to the components in some technological formulations, whereas the latter appeared as monophasic systems. Therefore, it may be assumed that the behavior in multicomponent formulations is the result of a complex interplay of synergistic interactions between the different components that will require revisiting when new, more eco-sustainable ingredients are considered.
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Particles adsorbed to fluid interfaces are ubiquitous in industry, nature or life. The wide range of properties arising from the assembly of particles at fluid interface has stimulated an intense research activity on shed light to the most fundamental physico-chemical aspects of these systems. These include the mechanisms driving the equilibration of the interfacial layers, trapping energy, specific inter-particle interactions and the response of the particle-laden interface to mechanical perturbations and flows. The understanding of the physico-chemistry of particle-laden interfaces becomes essential for taking advantage of the particle capacity to stabilize interfaces for the preparation of different dispersed systems (emulsions, foams or colloidosomes) and the fabrication of new reconfigurable interface-dominated devices. This review presents a detailed overview of the physico-chemical aspects that determine the behavior of particles trapped at fluid interfaces. This has been combined with some examples of real and potential applications of these systems in technological and industrial fields. It is expected that this information can provide a general perspective of the topic that can be exploited for researchers and technologist non-specialized in the study of particle-laden interfaces, or for experienced researcher seeking new questions to solve.
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Coloides , Coloides/química , Emulsiones/químicaRESUMEN
Polyelectrolyte multilayered capsules (PEMUCs) obtained using the Layer-by-Layer (LbL) method have become powerful tools for different biomedical applications, which include drug delivery, theranosis or biosensing. However, the exploitation of PEMUCs in the biomedical field requires a deep understanding of the most fundamental bases underlying their assembly processes, and the control of their properties to fabricate novel materials with optimized ability for specific targeting and therapeutic capacity. This review presents an updated perspective on the multiple avenues opened for the application of PEMUCs to the biomedical field, aiming to highlight some of the most important advantages offered by the LbL method for the fabrication of platforms for their use in the detection and treatment of different diseases.
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The formation of coffee-ring deposits upon evaporation of sessile droplets containing mixtures of poly(diallyldimethylammonium chloride) (PDADMAC) and two different anionic surfactants were studied. This process is driven by the Marangoni stresses resulting from the formation of surface-active polyelectrolyte-surfactant complexes in solution and the salt arising from the release of counterions. The morphologies of the deposits appear to be dependent on the surfactant concentration, independent of their chemical nature, and consist of a peripheral coffee ring composed of PDADMAC and PDADMAC-surfactant complexes, and a secondary region of dendrite-like structures of pure NaCl at the interior of the residue formed at the end of the evaporation. This is compatible with a hydrodynamic flow associated with the Marangoni stress from the apex of the drop to the three-phase contact line for those cases in which the concentration of the complexes dominates the surface tension, whereas it is reversed when most of the PDADMAC and the complexes have been deposited at the rim and the bulk contains mainly salt.
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Polielectrolitos/química , Polietilenos/química , Compuestos de Amonio Cuaternario/química , Silicio/química , Tensoactivos/química , Tensión SuperficialRESUMEN
Particle-laden fluid/fluid interfaces are ubiquitous in academia and industry, which has fostered extensive research efforts trying to disentangle the physico-chemical bases underlying the trapping of particles to fluid/fluid interfaces as well as the properties of the obtained layers. The understanding of such aspects is essential for exploiting the ability of particles on the stabilization of fluid/fluid interface for the fabrication of novel interface-dominated devices, ranging from traditional Pickering emulsions to more advanced reconfigurable devices. This review tries to provide a general perspective of the physico-chemical aspects associated with the stabilization of interfaces by colloidal particles, mainly chemical isotropic spherical colloids. Furthermore, some aspects related to the exploitation of particle-laden fluid/fluid interfaces on the stabilization of emulsions and foams will be also highlighted. It is expected that this review can be used for researchers and technologist as an initial approach to the study of particle-laden fluid layers.
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Magnetic colloids adsorbed at a fluid interface are unique model systems to understand self-assembly in confined environments, both in equilibrium and out of equilibrium, with important potential applications. In this work the pearl-chain-like self-assembled structures of superparamagnetic colloids confined to a fluid-fluid interface under static and time-dependent actuations are investigated. On the one hand, it is found that the structures generated by static fields transform as the tilt angle of the field with the interface is increased, from 2D crystals to separated pearl-chains in a process that occurs through a controllable and reversible zip-like thermally activated mechanism. On the other hand, the actuation with precessing fields about the axis perpendicular to the interface induces dynamic self-assembled structures with no counterpart in non-confined systems, generated by the interplay of averaged magnetic interactions, interfacial forces, and hydrodynamics. Finally, how these dynamic structures can be used as remotely activated roller conveyors, able to transport passive colloidal cargos at fluid interfaces and generate parallel viscous flows is shown. The latter can be used in the mixture of adsorbed molecules and the acceleration of surface-chemical reactions, overcoming diffusion limitations.
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This work presents a simple methodology for coating small unilamellar liposomes bearing different degrees of positive charge with polyelectrolyte multilayers using the sequential layer-by-layer deposition method. The liposomes were made of mixtures of 1,2-dioleyl-sn-glycero-3-phosphocoline and dimethyl dioctadecyl ammonium bromide (DODAB) and coated by alternated layers of the sodium salt of poly(4-styrenesulfonate) (PSS) and poly(allylamine) (PAH) as polyanions and polycations, respectively. The results show that the zeta potential of the liposomes was not very sensitive to the mole fraction of DODAB in the membrane, XD, in the range 0.3 ≤ XD ≤ 0.8. We were able to coat the liposomes with up to four polymer bilayers. The growth of the capsule size was followed by dynamic light scattering, and in some cases, by cryo-transmission electron microscopy, with good agreement between both techniques. The thickness of the layers, measured from the hydrodynamic radius of the coated liposome, depends on the polyelectrolyte used, so that the PSS layers adopt a much more packaged conformation than the PAH layers. An interesting finding is that the PSS amount needed to reach the isoelectric point of the capsules increases linearly with the charge density of the bare liposomes, whereas the amount of PAH does not depend on it. As expected, the preparation of the multilayers has to be done in such a way that when the system is close to the isoelectric point, the capsules do not aggregate. For this, we dropped the polyelectrolyte solution quickly, stirred it fast, and used dilute liposome suspensions. The method is very flexible and not limited to liposomes or polyelectrolyte multilayers; also, coatings containing charged nanoparticles can be easily made. Once the liposomes have been coated, lipids can be easily eliminated, giving rise to polyelectrolyte nanocapsules (polyelectrosomes) with potential applications as drug delivery platforms.
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Liposomas , Nanopartículas , Cápsulas , Microscopía Electrónica de Transmisión , PolielectrolitosRESUMEN
The Layer-by-Layer (LbL) method is a well-established method for the assembly of nanomaterials with controlled structure and functionality through the alternate deposition onto a template of two mutual interacting molecules, e.g., polyelectrolytes bearing opposite charge. The current development of this methodology has allowed the fabrication of a broad range of systems by assembling different types of molecules onto substrates with different chemical nature, size, or shape, resulting in numerous applications for LbL systems. In particular, the use of soft colloidal nanosurfaces, including nanogels, vesicles, liposomes, micelles, and emulsion droplets as a template for the assembly of LbL materials has undergone a significant growth in recent years due to their potential impact on the design of platforms for the encapsulation and controlled release of active molecules. This review proposes an analysis of some of the current trends on the fabrication of LbL materials using soft colloidal nanosurfaces, including liposomes, emulsion droplets, or even cells, as templates. Furthermore, some fundamental aspects related to deposition methodologies commonly used for fabricating LbL materials on colloidal templates together with the most fundamental physicochemical aspects involved in the assembly of LbL materials will also be discussed.
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The inherent thermodynamic instability of liposomes during production and storage has limited their widespread applications. Therefore, a novel structure of food-grade nanoliposomes stabilized by a 3D organogel network within the bilayer shell was developed through the extrusion process and successfully applied to encapsulate vitamin D3. A huge flocculation and a significant reduction of zeta potential (-17 mV) were observed in control nanoliposomes (without the organogel shell) after 2 months of storage at 4 °C, while the sample with a gelled bilayer showed excellent stability with a particle diameter of 105 nm and a high negative zeta potential (-63.4 mV), even after 3 months. The development of spherical vesicles was confirmed by TEM. Interestingly, the gelled bilayer shell led to improved stability against osmotically active divalent salt ions. Electron paramagnetic resonance confirmed the higher rigidity of the shell bilayer upon gelation. The novel liposome offered a dramatic increase in encapsulation efficiency and loading of vitamin D3 compared to those of control.
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Colecalciferol , Liposomas , Tamaño de la PartículaRESUMEN
The aberrant gain of DNA methylation at CpG islands is frequently observed in colorectal tumours and may silence the expression of tumour suppressors such as MLH1. Current models propose that these CpG islands are targeted by de novo DNA methyltransferases in a sequence-specific manner, but this has not been tested. Using ectopically integrated CpG islands, here we find that aberrantly methylated CpG islands are subject to low levels of de novo DNA methylation activity in colorectal cancer cells. By delineating DNA methyltransferase targets, we find that instead de novo DNA methylation activity is targeted primarily to CpG islands marked by the histone modification H3K36me3, a mark associated with transcriptional elongation. These H3K36me3 marked CpG islands are heavily methylated in colorectal tumours and the normal colon suggesting that de novo DNA methyltransferase activity at CpG islands in colorectal cancer is focused on similar targets to normal tissues and not greatly remodelled by tumourigenesis.
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Neoplasias Colorrectales/genética , ADN (Citosina-5-)-Metiltransferasas/metabolismo , Metilación de ADN , Regulación Neoplásica de la Expresión Génica , Código de Histonas/genética , Carcinogénesis/genética , Línea Celular Tumoral , Secuenciación de Inmunoprecipitación de Cromatina , Colon/patología , Neoplasias Colorrectales/patología , Islas de CpG/genética , ADN (Citosina-5-)-Metiltransferasas/genética , Conjuntos de Datos como Asunto , Epigénesis Genética , Técnicas de Inactivación de Genes , Histonas/genética , Humanos , Regiones Promotoras Genéticas/genética , Transcripción GenéticaRESUMEN
The adsorption of mixtures formed by chitosan and sodium lauryl ether sulfate (SLES) at the water/vapor interface has been studied on the basis of their impact on the equilibrium surface tension of the interface, and the response of such an interface to mechanical deformations. The analysis of the surfactant binding to the chitosan chains evidenced that the chitosan-SLES solutions were mixtures of polyelectrolyte-surfactant complexes and a non-negligible amount of free surfactant molecules. The interfacial properties showed two well-differentiated regions for interfacial adsorption as a function of the SLES concentration: (i) at a low surfactant concentration, co-adsorption of chitosan and SLES occurs, and (ii) at high concentrations, the surface is mostly occupied by SLES molecules. This behavior may be interpreted in terms of a complex equilibration mechanism of the interfacial layers, where different coupled dynamic processes may be involved. Furthermore, the use of the time-concentration superposition principle has confirmed the different dynamic behaviors of the chitosan-SLES adsorption as a function of the SLES concentration. This work sheds light on some of the most fundamental bases governing the physico-chemical behavior of mixtures formed by a biopolymer and a surfactant, where their complex behavior is governed by an intricate balance of bulk and interfacial interactions.
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The understanding of the deposition of oppositely charged polyelectrolytes-surfactant mixtures onto solid surfaces presents a high interest in current days due to the recognized impact of the obtained layers on different industrial sectors and the performance of several consumer products (e.g. formulations of shampoos and hair conditioners). This results from the broad range of structures and properties that can present the mixed layers, which in most of the cases mirror the association process occurring between the polyelectrolyte chains and the oppositely charged surfactants in the bulk. Therefore, the understanding of the adsorption processes and characteristics of the adsorbed layers can be only attained from a careful examination of the self-assembly processes occurring in the solution. This review aims to contribute to the understanding of the interaction of polyelectrolyte-surfactant mixtures with solid surfaces, which is probably one of the most underexplored aspects of these type of systems. For this purpose, a comprehensive discussion on the correlations between the aggregates formed in the solutions and the deposition of the obtained complexes upon such association onto solid surfaces will be presented. This makes it necessary to take a closer look to the most important forces driving such processes.