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Self-sorting in functionalized dipeptide systems can be driven by the chirality of a single amino acid, both at a high pH in the micellar state and at a low pH in the gel state. The structures formed are affected to some degree by the relative concentrations of each component showing the complexity of such an approach. The structures underpinning the gel network are predefined by the micellar structures at a high pH. Here, we describe the systems prepared from two dipeptide-based gelators that differ only by the chirality of one of the amino acids. We provide firm evidence for self-sorting in the micellar and gel phases using small-angle neutron scattering and cryo-transmission electron microscopy (cryo-TEM), showing that complete self-sorting occurs across a range of relative concentrations.
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Dipeptídeos , Micelas , Dipeptídeos/química , Microscopia Eletrônica de Transmissão , Microscopia Crioeletrônica , AminoácidosRESUMO
The polymorphism of lipid aggregates has long attracted detailed study due to the myriad factors that determine the final mesophase observed. This study is driven by the need to understand mesophase behaviour for a number of applications, such as drug delivery and membrane protein crystallography. In the case of the latter, the role of the so-called 'sponge' (L3) mesophase has been often noted, but not extensively studied by itself. The L3 mesophase can be formed in monoolein/water systems on the addition of butanediol to water, which partitions the headgroup region of the membrane, and decreases its elastic moduli. Like cubic mesophases, it is bicontinuous, but unlike them, has no long-range translational symmetry. In our present study, we show that the formation of the L3 phase can delicately depend on the addition of dopant lipids to the mesophase. While electrostatically neutral molecules similar in shape to monoolein (DOPE, cholesterol) have little effect on the general mesophase behaviour, others (DOPC, DDM) significantly reduce the composition at which it can form. Additionally, we show that by combining cholesterol with the anionic lipid DOPG, it is possible to form the largest stable L3 mesophases observed to date, with characteristic lengths over 220 Å.
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Phase transitions of proteins are strongly influenced by surface chemical modifications or mutations. Human γD-crystallin (HGD) single-mutants have been extensively studied because they are associated with the onset of juvenile cataract. However, they have also provided a rich library of molecules to examine how specific inter-protein interactions direct protein assembly, providing new insights and valuable experimental data for coarse-grained patchy-particle models. Here, we demonstrate that the addition of new inter-protein interactions by mutagenesis is additive and increases the number and variety of condensed phases formed by proteins. When double mutations incorporating two specific single point mutations are made, the properties of both single mutations are retained in addition to the formation of a new condensed phase. We find that the HGD double-mutant P23VC110M self-assembles into spherical particles with retrograde solubility, orthorhombic crystals, and needle/plate shape crystals, while retaining the ability to undergo liquid-liquid phase separation. This rich polymorphism is only partially predicted by the experimental data on the constituent single mutants. We also report a previously un-characterized amorphous protein particle, with unique properties that differ from those of protein spherulites, protein particulates previously described. The particles we observe are amorphous, reversible with temperature, tens of microns in size, and perfectly spherical. When they are grown on pristine surfaces, they appear to form by homogeneous nucleation, making them unique, and we believe a new form of protein condensate. This work highlights the challenges in predicting protein behavior, which has frustrated rational assembly and crystallization but also provides rich data to develop new coarse-grained models to explain the observed polymorphism.
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Mutação Puntual , Humanos , Anisotropia , Transição de Fase , Mutação , TemperaturaRESUMO
To accurately represent the morphological and elastic properties of a human red blood cell, Fu et al. [Fu et al., Lennard-Jones type pair-potential method for coarse-grained lipid bilayer membrane simulations in LAMMPS, 2017, 210, 193-203] recently developed a coarse-grained molecular dynamics model with particular detail in the membrane. However, such a model accrues an extremely high computational cost for whole-cell simulation when assuming an appropriate length scaling - that of the bilayer thickness. To date, the model has only simulated "miniature" cells in order to circumvent this, with the a priori assumption that these miniaturised cells correctly represent their full-sized counterparts. The present work assesses the validity of this approach, by testing the scale invariance of the model through simulating cells of various diameters; first qualitatively in their shape evolution, then quantitatively by measuring their bending rigidity through fluctuation analysis. Cells of diameter of at least 0.5 µm were able to form the characteristic biconcave shape of human red blood cells, though smaller cells instead equilibrated to bowl-shaped stomatocytes. Thermal fluctuation analysis showed the bending rigidity to be constant over all cell sizes tested, and consistent between measurements on the whole-cell and on a planar section of bilayer. This is as expected from the theory on both counts. Therefore, we confirm that the evaluated model is a good representation of a full-size RBC when the model diameter is ≥0.5 µm, in terms of the morphological and mechanical properties investigated.
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Eritrócitos , Bicamadas Lipídicas , Simulação por Computador , Eritrócitos/metabolismo , Humanos , Bicamadas Lipídicas/metabolismo , Modelos MolecularesRESUMO
The material properties of a gel are determined by the underpinning network that immobilises the solvent. When gels are formed by the self-assembly of small molecules into a so-called low molecular weight gel, the network is the result of the molecules forming one-dimensional objects such as fibres or nanotubes which entangle or otherwise cross-link to form a three-dimensional network. Characterising the one-dimensional objects and the network is difficult. Many conventional techniques rely on drying to probe the network, which often leads to artefacts. An effective tool to probe the gel in the solvated state is small angle scattering. Both small angle X-ray scattering (SAXS) and small angle neutron scattering (SANS) can be used. Here, we discuss these approaches and provide a tutorial review to describe how these approaches work, what opportunities there are and how the data treatment should be approached. We aim to show the power of this approach and provide enabling information to make them accessible to the non-specialist.
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While proteins have been treated as particles with a spherically symmetric interaction, of course in reality, the situation is rather more complex. A simple step toward higher complexity is to treat the proteins as non-spherical particles and that is the approach we pursue here. We investigate the phase behavior of the enhanced green fluorescent protein (eGFP) under the addition of a non-adsorbing polymer, polyethylene glycol. From small angle x-ray scattering, we infer that the eGFP undergoes dimerization and we treat the dimers as spherocylinders with aspect ratio L/D - 1 = 1.05. Despite the complex nature of the proteins, we find that the phase behavior is similar to that of hard spherocylinders with an ideal polymer depletant, exhibiting aggregation and, in a small region of the phase diagram, crystallization. By comparing our measurements of the onset of aggregation with predictions for hard colloids and ideal polymers [S. V. Savenko and M. Dijkstra, J. Chem. Phys. 124, 234902 (2006) and Lo Verso et al., Phys. Rev. E 73, 061407 (2006)], we find good agreement, which suggests that the behavior of the eGFP is consistent with that of hard spherocylinders and ideal polymers.
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Coloides , Polímeros , Agregados Proteicos , Proteínas , Coloides/química , Cristalização , Polímeros/química , Proteínas/químicaRESUMO
Evaporation of a particle laden sessile drop can lead to complex surface patterns with structural hierarchy. Most commonly, the dispersed particles are inert. We have recently reported that when the sessile drop contains reactive ZnO nanoparticles, solidified Bénard-Marangoni (BM) cells with dendritic micromorphology were formed in the residual surface pattern from in situ-generated nanoclusters. Here, we report the effect of substrate chemistry on the residual pattern from the evaporation of nanofluids containing ZnO particles dispersed in a mixture of cyclohexane and isobutylamine, by comparing three different substrates: glass, silicon, and hydrophilized silicon. In particular, we performed a quantitative analysis of the BM cell size, distribution, and the cell morphological characteristics via the fractal dimension analysis. We find that the size dimension λBM of the dendritic Bénard-Marangoni cells varied on the different substrates, attributed to their different hydrophilicity and affinity for water molecules, evident from the different polar components γP in their surface free energy from the Owens-Wendt analysis. The average BM cell size was the smallest for the glass substrate (λBM = 289 µm) and comparable for the unmodified and UV/ozone-treated silicon wafers (with λBM = 466 and 423 µm, respectively). The fractal dimension analysis provided a quantitative description of the BM cells with complex structural hierarchy, highlighting the differences in the geometric features of the surface patterns resulting from different substrate chemistry. We also found that the fractal dimensions depended on the BM cell size, attributing it to two different regimes: the growing fractals and the maturing fractals.
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Surface structures with tailored morphologies can be readily delivered by the evaporation-induced self-assembly process. It has been recently demonstrated that ZnO nanorods could undergo rapid chemical and morphological transformation into 3D complex structures of Zn(OH)2 nanofibers as a droplet of ZnO nanofluid dries on the substrate via a mechanism very different from that observed in the coffee ring effect. Here, we have investigated how the crystallinity and morphology of ZnO nanoparticles would affect the ultimate pattern formation. Three ZnO particles differing in size and shape were used, and their crystal structures were characterized by powder X-ray diffraction (XRD) and transmission electron microscopy (TEM). Their dispersions were prepared by sonication in a mixture of isobutylamine and cyclohexane. Residual surface patterns were created by drop casting a droplet of the nanofluid on a silicon substrate. The residual surface patterns were analyzed by scanning electron microscopy (SEM) and microfocus grazing incidence X-ray diffraction (µGIXRD). Nanofluid droplets of the in-house synthesized ZnO nanoparticles resulted in residual surface patterns consisting of Zn(OH)2 nanofibers. However, when commercially acquired ZnO powders composed of crystals with various shapes and sizes were used as the starting material, Zn(OH)2 fibers were found covered by ZnO crystal residues that did not fully undergo the dissolution and recrystallization process during evaporation. The difference in the solubility of ZnO nanoparticles was linked to the difference in their crystallinity, as assessed using the Scherrer equation analysis of their XRD Bragg peaks. Our results show that the morphology of the ultimate residual pattern from evaporation of ZnO nanofluids can be controlled by varying the crystallinity of the starting ZnO nanoparticles which affects the nanoparticle dissolution process during evaporation.
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We propose a novel microfluidic "opposed-flow" geometry in which the continuous fluid phase is fed into a junction in a direction opposite to the dispersed phase. This pulls out the dispersed phase into a micron-sized jet, which decays into micron-sized droplets. As the driving pressure is tuned to a critical value, the jet radius vanishes as a power law down to sizes below 1 µm. By contrast, the conventional "coflowing" junction leads to a first order jetting transition, in which the jet disappears at a finite radius of several µm, to give way to a "dripping" state, resulting in much larger droplets. We demonstrate the effectiveness of our method by producing the first microfluidic silicone oil emulsions with a sub micron particle radius, and utilize these droplets to produce colloidal clusters.
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As the use of low-molecular-weight gelators (LMWGs) as components in single and multicomponent systems for optoelectronic and solar cell applications increases, so does the need for more functional gelators. There are relatively few examples of p-type gelators that can be used in such systems. Here, the synthesis and characterization of three amino-acid-functionalized p-type gelators based on terthiophene, tetrathiafulvalene, and oligo(phenylenevinylene) are described. The cores of these molecules are already used as electron donors in optoelectronic applications. These newly designed molecules can gel water to form highly organized structures, which can be dried into thin films that show p-type behavior.
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Aminoácidos/química , Hidrogel de Polietilenoglicol-Dimetacrilato/química , Hidrogel de Polietilenoglicol-Dimetacrilato/síntese química , Estrutura Molecular , Peso MolecularRESUMO
We have developed norland optical adhesive (NOA) flow focusing devices, making use of the excellent solvent compatibility and surface properties of NOA to generate micron scale oil-in-water emulsions with polydispersities as low as 5%. While current work on microfluidic oil-in-water emulsification largely concerns the production of droplets with sizes on the order of 10s of micrometres, large enough that Brownian motion is negligible, our NOA devices can produce droplets with radii ranging from 2 µm to 12 µm. To demonstrate the utility of these emulsions as colloidal model systems we produce fluorescently labelled polydimethylsiloxane droplets suitable for particle resolved studies with confocal microscopy. We analyse the structure of the resulting emulsion in 3D using coordinate tracking and the topological cluster classification and reveal a new mono-disperse thermal system.
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A tetra(aniline)-based cationic amphiphile, TANI-NHC(O)C5H10N(CH3)3(+)Br(-) (TANI-PTAB) was synthesized, and its emeraldine base (EB) state was found to self-assemble into nanowires in aqueous solution. The observed self-assembly is described by an isodesmic model, as shown by temperature-dependent UV-vis investigations. Linear dichroism (LD) studies, combined with computational modeling using time-dependent density functional theory (TD-DFT), suggests that TANI-PTAB molecules are ordered in an antiparallel arrangement within nanowires, with the long axis of TANI-PTAB arranged perpendicular to the nanowire long axis. Addition of either S- or R- camphorsulfonic acid (CSA) to TANI-PTAB converted TANI to the emeraldine salt (ES), which retained the ability to form nanowires. Acid doping of TANI-PTAB had a profound effect on the nanowire morphology, as the CSA counterions' chirality translated into helical twisting of the nanowires, as observed by circular dichroism (CD). Finally, the electrical conductivity of CSA-doped helical nanowire thin films processed from aqueous solution was 2.7 mS cm(-1). The conductivity, control over self-assembled 1D structure and water-solubility demonstrate these materials' promise as processable and addressable functional materials for molecular electronics, redox-controlled materials and sensing.
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A macroscopically oriented double diamond inverse bicontinuous cubic phase (QII(D)) of the lipid glycerol monooleate is reversibly converted into a gyroid phase (QII(G)). The initial QII(D) phase is prepared in the form of a film coating the inside of a capillary, deposited under flow, which produces a sample uniaxially oriented with a ⟨110⟩ axis parallel to the symmetry axis of the sample. A transformation is induced by replacing the water within the capillary tube with a solution of poly(ethylene glycol), which draws water out of the QII(D) sample by osmotic stress. This converts the QII(D) phase into a QII(G) phase with two coexisting orientations, with the ⟨100⟩ and ⟨111⟩ axes parallel to the symmetry axis, as demonstrated by small-angle X-ray scattering. The process can then be reversed, to recover the initial orientation of QII(D) phase. The epitaxial relation between the two oriented mesophases is consistent with topology-preserving geometric pathways that have previously been hypothesized for the transformation. Furthermore, this has implications for the production of macroscopically oriented QII(G) phases, in particular with applications as nanomaterial templates.
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We have studied the effect of pressure and temperature on the location of the pivotal surface in a lipid inverse bicontinuous gyroid cubic phase (Q(G)(II)), described by the area at the pivotal surface (An), the volume between the pivotal surface and the bilayer midplane (Vn), and the molecular volume of the lipid (V). Small angle X-ray scattering (SAXS) was used to measure the swelling behaviour of the lipid, monolinolein, as a function of pressure and temperature, and the data were fitted to two different geometric models: the parallel interface model (PIM), and the constant mean curvature model (CMCM). The results show that an increase in temperature leads to a shift in the location of the pivotal surface towards the bilayer midplane, whilst an increase in pressure causes the pivotal surface to move towards the interfacial region. In addition, we describe the relevance of An, Vn and V for modeling the energetics of curved mesophases with specific reference to the mean curvature at the pivotal surface and discuss the significance of this parameter for modelling the energetics of curved mesophases.
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We demonstrate a method by which we can produce an oriented film of an inverse bicontinuous cubic phase (Q(II)(D)) formed by the lipid monoolein (MO). By starting with the lipid as a disordered precursor (the L(3) phase) in the presence of butanediol, we can obtain a film of the Q(II)(D) phase showing a high degree of in-plane orientation by controlled dilution of the sample under shear within a linear flow cell. We demonstrate that the direction of orientation of the film is different from that found in the oriented bulk material that we have reported previously; therefore, we can now reproducibly form Q(II)(D) samples oriented with either the [110] or the [100] axis aligned in the flow direction depending on the method of preparation. The deposition of MO as a film, via a moving fluid-air interface that leaves a coating of MO in the L(3) phase on the capillary wall, leads to a sample in the [110] orientation. This contrasts with the bulk material that we have previously demonstrated to be oriented in the [100] direction, arising from flow producing an oriented bulk slug of material within the capillary tube. The bulk sample contains significant amounts of residual butanediol, which can be estimated from the lattice parameter of the Q(II)(D) phase obtained. The sample orientation and lattice parameters are determined from synchrotron small-angle X-ray scattering patterns and confirmed by simulations. This has potential applications in the production of template materials and the growth of protein crystals for crystallography as well as deepening our understanding of the mechanisms underlying the behavior of lyotropic liquid-crystal phases.
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Glicerídeos/química , Butileno Glicóis/química , Modelos Moleculares , Conformação Molecular , Espalhamento a Baixo Ângulo , Fatores de Tempo , Difração de Raios XRESUMO
Direct measurements to determine the degree of surface coverage of nanoparticles by functional moieties are rare, with current strategies requiring a high level of expertise and expensive equipment. Here, a practical method to determine the ratio of the volume of the functionalisation layer to the particle volume based on measuring the refractive index of nanoparticles in suspension is proposed. As a proof of concept, this technique is applied to poly(methyl methacrylate) (PMMA) nanoparticles and semicrystalline carbon dots functionalised with different surface moieties, yielding refractive indices that are commensurate to those from previous literature and Mie theory. In doing so, it is demonstrated that this technique is able to optically detect differences in surface functionalisation or composition of nanometre-sized particles. This non-destructive and rapid method is well-suited for in situ industrial particle characterisation and biological applications.
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We demonstrate the formation of a macroscopically oriented inverse bicontinuous cubic (Q(II)) lipid phase from a sponge (L(3)) phase by controlled hydration during shear flow. The L(3) phase was the monoolein/butanediol/water system; the addition of water reduces the butanediol concentration, inducing the formation of a diamond (Q(II)(D)) cubic phase, which is oriented by the shear flow. The phenomenon was reproduced in both capillary and Couette geometries, indicating that this represents a robust general route for the production of highly aligned bulk Q(II) samples, with applications in nanomaterial templating and protein research.
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Lipídeos/química , Butileno Glicóis/química , Glicerídeos/química , Modelos Moleculares , Transição de Fase , Água/químicaRESUMO
We demonstrate a solution method that allows both elongation rate and average fibril length of assembling amyloid fibrils to be estimated. The approach involves acquisition of real-time neutron scattering data during the initial stages of seeded growth, using contrast matched buffer to make the seeds effectively invisible to neutrons. As deuterated monomers add on to the seeds, the labelled growing ends give rise to scattering patterns that we model as cylinders whose increase in length with time gives an elongation rate. In addition, the absolute intensity of the signal can be used to determine the number of growing ends per unit volume, which in turn provides an estimate of seed length. The number of ends did not change significantly during elongation, demonstrating that any spontaneous or secondary nucleation was not significant compared with growth on the ends of pre-existing fibrils, and in addition providing a method of internal validation for the technique. Our experiments on initial growth of alpha synuclein fibrils using 1.2 mg ml-1 seeds in 2.5 mg ml-1 deuterated monomer at room temperature gave an elongation rate of 6.3 ± 0.5 Å min-1, and an average seed length estimate of 4.2 ± 1.3 µm.
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The field of drug-membrane interactions is one that spans a wide range of scientific disciplines, from synthetic chemistry, through biophysics to pharmacology. Cell membranes are complex dynamic systems whose structures can be affected by drug molecules and in turn can affect the pharmacological properties of the drugs being administered. In this tutorial review we aim to provide a guide for those new to the area of drug-membrane interactions and present an introduction to areas of this topic which need to be considered. We address the lipid composition and structure of the cell membrane and comment on the physical forces present in the membrane which may impact on drug interactions. We outline methods by which drugs may cross or bind to this membrane, including the well understood passive and active transport pathways. We present a range of techniques which may be used to study the interactions of drugs with membranes both in vitro and in vivo and discuss the advantages and disadvantages of these techniques and highlight new methods being developed to further this field.
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Membrana Celular , Interações Medicamentosas/fisiologia , Animais , Transporte Biológico/fisiologia , Membrana Celular/efeitos dos fármacos , Membrana Celular/fisiologia , Humanos , Modelos Biológicos , Análise Espectral/métodosRESUMO
The effects of biological buffers on lipids have not been fully investigated because of the long-standing assumption that these buffers are too hydrophilic to substantially interact with the lipid membrane. We present evidence that for some buffers, this is not necessarily the case. Our research points toward a membrane softening effect caused by the buffer molecules interacting with the headgroup region of the lipid. Changes in the elastic properties of the membrane are known to control membrane protein behavior; this work serves as a warning for the design of assays utilizing model membranes in the presence of buffers.