86 results found
Tyler AII, Greenfield JL, Seddon JM, et al., 2019, Coupling Phase Behavior of Fatty Acid Containing Membranes to Membrane Bio-Mechanics, FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY, Vol: 7, ISSN: 2296-634X
Weber CC, Brooks NJ, Castiglione F, et al., 2019, On the structural origin of free volume in 1-alkyl-3-methylimidazolium ionic liquid mixtures: a SAXS and 129Xe NMR study., Physical Chemistry Chemical Physics, Vol: 21, Pages: 5999-6010, ISSN: 1463-9076
Ionic liquid (IL) mixtures enable the design of fluids with finely tuned structural and physicochemical properties for myriad applications. In order to rationally develop and design IL mixtures with the desired properties, a thorough understanding of the structural origins of their physicochemical properties and the thermodynamics of mixing needs to be developed. To elucidate the structural origins of the excess molar volume within IL mixtures containing ions with different alkyl chain lengths, 3 IL mixtures containing 1-alkyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide ILs have been explored in a joint small angle X-ray scattering (SAXS) and 129Xe NMR study. The apolar domains of the IL mixtures were shown to possess similar dimensions to the largest alkyl chain of the mixture with the size evolution determined by whether the shorter alkyl chain was able to interact with the apolar domain. 129Xe NMR results illustrated that the origin of excess molar volume in these mixtures was due to fluctuations within these apolar domains arising from alkyl chain mismatch, with the formation of a greater number of smaller voids within the IL structure. These results indicate that free volume effects for these types of mixtures can be predicted from simple considerations of IL structure and that the structural basis for the formation of excess molar volume in these mixtures is substantially different to IL mixtures formed of different types of ions.
Woodcock EM, Girvan P, Eckert J, et al., Measuring intracellular viscosity in conditions of hypergravity, Biophysical Journal, ISSN: 0006-3495
Gravity-sensitive cellular responses are regularly observed in both specialised and non-specialised cells. One potential mechanism for this sensitivity is a changing viscosity of the intracellular organelles. Here, we report a novel viscosity-sensitive molecular rotor based on meso-substituted boron-dipyrrin (BODIPY) used to investigate the response of viscosity of cellular membranes to hypergravity conditions created at the Large Diameter Centrifuge at the European Space Agency Technology Centre ESA-ESTEC. Mouse osteoblastic (MC3T3-E1) and endothelial (HUVEC) cell lines were tested and an increase in viscosity was found with increasing hypergravity loading. This response is thought to be primarily biologically driven, with the potential for a small, instantaneous physical mechanism also contributing to the observed effect. This work provides the first quantitative data for cellular viscosity changes under hypergravity, up to 15 g.
Khan H, Seddon JM, Law RV, et al., 2019, Effect of glycerol with sodium chloride on the Krafft point of sodium dodecyl sulfate using surface tension, JOURNAL OF COLLOID AND INTERFACE SCIENCE, Vol: 538, Pages: 75-82, ISSN: 0021-9797
Brady RA, Kaufhold WT, Brooks NJ, et al., 2019, Flexibility defines structure in crystals of amphiphilic DNA nanostars, JOURNAL OF PHYSICS-CONDENSED MATTER, Vol: 31, ISSN: 0953-8984
Menny A, Serna M, Boyd C, et al., 2018, CryoEM reveals how the complement membrane attack complex ruptures lipid bilayers, Nature Communications, Vol: 9, ISSN: 2041-1723
The membrane attack complex (MAC) is one of the immune system’s first responders. Complement proteins assemble on target membranes to form pores that lyse pathogens and impact tissue homeostasis of self-cells. How MAC disrupts the membrane barrier remains unclear. Here we use electron cryo-microscopy and flicker spectroscopy to show that MAC interacts with lipid bilayers in two distinct ways. Whereas C6 and C7 associate with the outer leaflet and reduce the energy for membrane bending, C8 and C9 traverse the bilayer increasing membrane rigidity. CryoEM reconstructions reveal plasticity of the MAC pore and demonstrate how C5b6 acts as a platform, directing assembly of a giant β-barrel whose structure is supported by a glycan scaffold. Our work provides a structural basis for understanding how β-pore forming proteins breach the membrane and reveals a mechanism for how MAC kills pathogens and regulates cell functions.
Trantidou T, Friddin M, Gan KB, et al., 2018, Mask-free laser lithography for rapid and low-cost microfluidic device fabrication, Analytical Chemistry, Vol: 90, Pages: 13915-13921, ISSN: 0003-2700
Microfluidics has become recognized as a powerful platform technology associated with a constantly increasing array of applications across the life sciences. This surge of interest over recent years has led to an increased demand for microfluidic chips, resulting in more time being spent in the cleanroom fabricating devices using soft lithography—a slow and expensive process that requires extensive materials, training and significant engineering resources. This bottleneck limits platform complexity as a byproduct of lengthy delays between device iterations and affects the time spent developing the final application. To address this problem, we report a new, rapid, and economical approach to microfluidic device fabrication using dry resist films to laminate laser cut sheets of acrylic. We term our method laser lithography and show that our technique can be used to engineer 200 μm width channels for assembling droplet generators capable of generating monodisperse water droplets in oil and micromixers designed to sustain chemical reactions. Our devices offer high transparency, negligible device to device variation, and low X-ray background scattering, demonstrating their suitability for real-time X-ray-based characterization applications. Our approach also requires minimal materials and apparatus, is cleanroom free, and at a cost of around $1.00 per chip could significantly democratize device fabrication, thereby increasing the interdisciplinary accessibility of microfluidics.
Brady RA, Brooks NJ, Foderà V, et al., 2018, Amphiphilic-DNA platform for the design of crystalline frameworks with programmable structure and functionality, Journal of the American Chemical Society, Vol: 140, Pages: 15384-15392, ISSN: 1520-5126
The reliable preparation of functional, ordered, nanostructured frameworks would be a game changer for many emerging technologies, from energy storage to nanomedicine. Underpinned by the excellent molecular recognition of nucleic acids, along with their facile synthesis and breadth of available functionalizations, DNA nanotechnology is widely acknowledged as a prime route for the rational design of nanostructured materials. Yet, the preparation of crystalline DNA frameworks with programmable structure and functionality remains a challenge. Here we demonstrate the potential of simple amphiphilic DNA motifs, dubbed "C-stars", as a versatile platform for the design of programmable DNA crystals. In contrast to all-DNA materials, in which structure depends on the precise molecular details of individual building blocks, the self-assembly of C-stars is controlled uniquely by their topology and symmetry. Exploiting this robust self-assembly principle, we design a range of topologically identical, but structurally and chemically distinct C-stars that following a one-pot reaction self-assemble into highly porous, functional, crystalline frameworks. Simple design variations allow us to fine-tune the lattice parameter and thus control the partitioning of macromolecules within the frameworks, embed responsive motifs that can induce isothermal disassembly, and include chemical moieties to capture target proteins specifically and reversibly.
Miller RM, Cabral J, Robles E, et al., 2018, Crystallisation of sodium dodecyl sulfate–water micellar solutions with structurally similar additives: counterion variation, CrystEngComm, Vol: 20, Pages: 6834-6843, ISSN: 1466-8033
The effects of a series of structurally similar sodium dodecyl sulfate (SDS) additives on the crystallisation of SDS–water micellar solutions were investigated using a combination of differential scanning calorimetry, dynamic light scattering, optical microscopy and inductively coupled plasma optical emission spectroscopy. Seven different counterions were chosen from groups 1 and 2 of the periodic table to replace the sodium on SDS: LDS, (SDS), KDS, RbDS, CsDS, Mg(DS)2, Ca(DS)2 and Sr(DS)2. Two representative temperature profileswere employed – linear cooling ramps at rate of 0.5 °C min−1 to determine near-equilibrium kinetics and transitions and isothermal holds at 6 °C to elucidate morphological changes. Crystallisation of the reference solution 20% SDS–H2O with 0.25, 1.0 and 2.5% additive was generally promoted or inhibited even at the lowest concentrations. Melting points however remained largely unchanged, suggesting that the additives predominantly had a kinetic rather than thermodynamic effect. ICP-OES measurements for the solutions containing 1% additive indicated that most of the additives were integrated into the SDS crystals which was reflected by morphological changes, including the formation of hexagonal and oval shaped crystals. Our results both quantify and provide a morphological insight into the effect of a series of additives on the crystallisation of micellar SDS solutions, which can readily form due to preferential Na exchange.
Barlow N, Kusumaatmaja H, Salehi-Reyhani A, et al., 2018, Measuring bilayer surface energy and curvature in asymmetric droplet interface bilayers, Journal of the Royal Society Interface, Vol: 15, ISSN: 1742-5662
For the past decade, droplet interface bilayers (DIBs) have had an increased prevalence in biomolecular and biophysical literature. However, much of the underlying physics of these platforms is poorly characterized. To further our understanding of these structures, lipid membrane tension on DIB membranes is measured by analysing the equilibrium shape of asymmetric DIBs. To this end, the morphology of DIBs is explored for the first time using confocal laser scanning fluorescence microscopy. The experimental results confirm that, in accordance with theory, the bilayer interface of a volume-asymmetric DIB is curved towards the smaller droplet and a lipid-asymmetric DIB is curved towards the droplet with the higher monolayer surface tension. Moreover, the DIB shape can be exploited to measure complex bilayer surface energies. In this study, the bilayer surface energy of DIBs composed of lipid mixtures of phosphatidylgylcerol (PG) and phosphatidylcholine are shown to increase linearly with PG concentrations up to 25%. The assumption that DIB bilayer area can be geometrically approximated as a spherical cap base is also tested, and it is discovered that the bilayer curvature is negligible for most practical symmetric or asymmetric DIB systems with respect to bilayer area.
Girvan P, Teng X, Brooks NJ, et al., 2018, Redox Kinetics of the Amyloid-beta-Cu Complex and Its Biological Implications, BIOCHEMISTRY, Vol: 57, Pages: 6228-6233, ISSN: 0006-2960
Holme MN, Rana S, Barriga H, et al., 2018, A robust liposomal platform for direct colorimetric detection of sphingomyelinase enzyme and inhibitors, ACS Nano, Vol: 12, Pages: 8197-8207, ISSN: 1936-0851
The enzyme sphingomyelinase (SMase) is an important biomarker for several diseases such as Niemann Pick’s, atherosclerosis, multiple sclerosis, and HIV. We present a two-component colorimetric SMase activity assay that is more sensitive and much faster than currently available commercial assays. Herein, SMase-triggered release of cysteine from a sphingomyelin (SM)-based liposome formulation with 60 mol % cholesterol causes gold nanoparticle (AuNP) aggregation, enabling colorimetric detection of SMase activities as low as 0.02 mU/mL, corresponding to 1.4 pM concentration. While the lipid composition offers a stable, nonleaky liposome platform with minimal background signal, high specificity toward SMase avoids cross-reactivity of other similar phospholipases. Notably, use of an SM-based liposome formulation accurately mimics the natural in vivo substrate: the cell membrane. We studied the physical rearrangement process of the lipid membrane during SMase-mediated hydrolysis of SM to ceramide using small- and wide-angle X-ray scattering. A change in lipid phase from a liquid to gel state bilayer with increasing concentration of ceramide accounts for the observed increase in membrane permeability and consequent release of encapsulated cysteine. We further demonstrated the effectiveness of the sensor in colorimetric screening of small-molecule drug candidates, paving the way for the identification of novel SMase inhibitors in minutes. Taken together, the simplicity, speed, sensitivity, and naked-eye readout of this assay offer huge potential in point-of-care diagnostics and high-throughput drug screening.
Brooker HR, Gyamfi IA, Wieckowska A, et al., 2018, A novel live-cell imaging system reveals a reversible hydrostatic pressure impact on cell-cycle progression, JOURNAL OF CELL SCIENCE, Vol: 131, ISSN: 0021-9533
Bolognesi G, Friddin MS, Salehi-Reyhani S, et al., 2018, Sculpting and fusing biomimetic vesicle networks using optical tweezers, Nature Communications, Vol: 9, ISSN: 2041-1723
Constructing higher-order vesicle assemblies has discipline-spanning potential from responsive soft-matter materials to artificial cell networks in synthetic biology. This potential is ultimately derived from the ability to compartmentalise and order chemical species in space. To unlock such applications, spatial organisation of vesicles in relation to one another must be controlled, and techniques to deliver cargo to compartments developed. Herein, we use optical tweezers to assemble, reconfigure and dismantle networks of cell-sized vesicles that, in different experimental scenarios, we engineer to exhibit several interesting properties. Vesicles are connected through double-bilayer junctions formed via electrostatically controlled adhesion. Chemically distinct vesicles are linked across length scales, from several nanometres to hundreds of micrometres, by axon-like tethers. In the former regime, patterning membranes with proteins and nanoparticles facilitates material exchange between compartments and enables laser-triggered vesicle merging. This allows us to mix and dilute content, and to initiate protein expression by delivering biomolecular reaction components.
Karamdad K, Hindley J, Friddin MS, et al., 2018, Engineering thermoresponsive phase separated vesicles formed via emulsion phase transfer as a content-release platform, Chemical Science, Vol: 9, Pages: 4851-4858, ISSN: 2041-6520
Giant unilamellar vesicles (GUVs) are a well-established tool for the study of membrane biophysics and are increasingly used as artificial cell models and functional units in biotechnology. This trend is driven by the development of emulsion-based generation methods such as Emulsion Phase Transfer (EPT), which facilitates the encapsulation of almost any water-soluble compounds (including biomolecules) regardless of size or charge, is compatible with droplet microfluidics, and allows GUVs with asymmetric bilayers to be assembled. However, the ability to control the composition of membranes formed via EPT remains an open question; this is key as composition gives rise to an array of biophysical phenomena which can be used to add functionality to membranes. Here, we evaluate the use of GUVs constructed via this method as a platform for phase behaviour studies and take advantage of composition-dependent features to engineer thermally-responsive GUVs. For the first time, we generate ternary GUVs (DOPC/DPPC/cholesterol) using EPT, and by compensating for the lower cholesterol incorporation efficiencies, show that these possess the full range of phase behaviour displayed by electroformed GUVs. As a demonstration of the fine control afforded by this approach, we demonstrate release of dye and peptide cargo when ternary GUVs are heated through the immiscibility transition temperature, and show that release temperature can be tuned by changing vesicle composition. We show that GUVs can be individually addressed and release triggered using a laser beam. Our findings validate EPT as a suitable method for generating phase separated vesicles and provide a valuable proof-of-concept for engineering content release functionality into individually addressable vesicles, which could have a host of applications in the development of smart synthetic biosystems.
Slatter DA, Percy CL, Allen-Redpath K, et al., 2018, Enzymatically oxidized phospholipids restore thrombin generation in coagulation factor deficiencies, JCI INSIGHT, Vol: 3, ISSN: 2379-3708
Barlow NE, Bolognesi G, Haylock S, et al., 2017, Rheological Droplet Interface Bilayers (rheo-DIBs): Probing the Unstirred Water Layer Effect on Membrane Permeability via Spinning Disk Induced Shear Stress, Scientific Reports, Vol: 7, ISSN: 2045-2322
A new rheological droplet interface bilayer (rheo-DIB) device is presented as a tool to apply shear stress on biological lipid membranes. Despite their exciting potential for affecting high-throughput membrane translocation studies, permeability assays conducted using DIBs have neglected the effect of the unstirred water layer (UWL). However as demonstrated in this study, neglecting this phenomenon can cause significant underestimates in membrane permeability measurements which in turn limits their ability to predict key processes such as drug translocation rates across lipid membranes. With the use of the rheo-DIB chip, the effective bilayer permeability can be modulated by applying shear stress to the droplet interfaces, inducing flow parallel to the DIB membranes. By analysing the relation between the effective membrane permeability and the applied stress, both the intrinsic membrane permeability and UWL thickness can be determined for the first time using this model membrane approach, thereby unlocking the potential of DIBs for undertaking diffusion assays. The results are also validated with numerical simulations.
de Bruin A, Friddin MS, Elani Y, et al., 2017, A transparent 3D printed device for assembling droplet hydrogel bilayers (DHBs), RSC Advances, Vol: 7, Pages: 47796-47800, ISSN: 2046-2069
We report a new approach for assembling droplet hydrogel bilayers (DHBs) using a transparent 3D printed device. We characterise the transparency of our platform, confirm bilayer formation using electrical measurements and show that single-channel recordings can be obtained using our reusable rapid prototyped device. This method significantly reduces the cost and infrastructure required to develop devices for DHB assembly and downstream study.
Richens JL, Tyler AII, Barriga HMG, et al., 2017, Spontaneous charged lipid transfer between lipid vesicles, Scientific Reports, Vol: 7, ISSN: 2045-2322
An assay to study the spontaneous charged lipid transfer between lipid vesicles is described. A donor/acceptor vesicle system is employed, where neutrally charged acceptor vesicles are uorescentlylabelled with the electrostatic membrane probe Fluoresceinphosphatidylethanolamine (FPE).Upon addition of charged donor vesicles, transfer of negatively charged lipid occurs, resulting ina uorescently detectable change in the membrane potential of the acceptor vesicles. Using this approach we have studied the transfer properties of a range of lipids, varying both the headgroup and the chain length. At the low vesicle concentrations chosen, the transfer follows a rst-order process where lipid monomers are transferred presumably through the aqueous solution phase from donor to acceptor vesicle. The rate of transfer decreases with increasing chain length which is consistent with energy models previously reported for lipid monomer vesicle interactions. Our assay improves on existing methods allowing the study of a range of unmodi ed lipids, continuous monitoring of transfer and simpli ed experimental procedures.
Cornell CE, McCarthy NLC, Levental KR, et al., 2017, Lengths of n-alcohols govern how Lo-Ld mixing temperatures shift in synthetic and cell-derived membranes, Biophysical Journal, Vol: 113, Pages: 1-13, ISSN: 1542-0086
A persistent challenge in membrane biophysics has been to quantitatively predict how membrane physical properties change upon addition of new amphiphiles (e.g., lipids, alcohols, peptides, or proteins) in order to assess whether the changes are large enough to plausibly result in biological ramifications. Because of their roles as general anesthetics, n-alcohols are perhaps the best-studied amphiphiles of this class. When n-alcohols are added to model and cell membranes, changes in membrane parameters tend to be modest. One striking exception is found in the large decrease in liquid-liquid miscibility transition temperatures (Tmix) observed when short-chain n-alcohols are incorporated into giant plasma membrane vesicles (GPMVs). Coexisting liquid-ordered and liquid-disordered phases are observed at temperatures below Tmix in GPMVs as well as in giant unilamellar vesicles (GUVs) composed of ternary mixtures of a lipid with a low melting temperature, a lipid with a high melting temperature, and cholesterol. Here, we find that when GUVs of canonical ternary mixtures are formed in aqueous solutions of short-chain n-alcohols (n ≤ 10), Tmix increases relative to GUVs in water. This shift is in the opposite direction from that reported for cell-derived GPMVs. The increase in Tmix is robust across GUVs of several types of lipids, ratios of lipids, types of short-chain n-alcohols, and concentrations of n-alcohols. However, as chain lengths of n-alcohols increase, nonmonotonic shifts in Tmix are observed. Alcohols with chain lengths of 10–14 carbons decrease Tmix in ternary GUVs of dioleoyl-PC/dipalmitoyl-PC/cholesterol, whereas 16 carbons increase Tmix again. Gray et al. observed a similar influence of the length of n-alcohols on the direction of the shift in Tmix. These results are consistent with a scenario in which the relative partitioning of n-alcohols between liquid-ordered and liquid-disordered phases evolves as the chain length of the n-alcohol increases.
Brooks NJ, Castiglione F, Doherty CM, et al., 2017, Linking the structures, free volumes, and properties of ionic liquid mixtures, Chemical Science, Vol: 8, Pages: 6359-6374, ISSN: 2041-6520
The formation of ionic liquid (IL) mixtures has been proposed as an approach to rationally fine-tune the physicochemical properties of ILs for a variety of applications. However, the effects of forming such mixtures on the resultant properties of the liquids are only beginning to be understood. Towards a more complete understanding of both the thermodynamics of mixing ILs and the effect of mixing these liquids on their structures and physicochemical properties, the spatial arrangement and free volume of IL mixtures containing the common [C4C1im]+ cation and different anions have been systematically explored using small angle X-ray scattering (SAXS), positron annihilation lifetime spectroscopy (PALS) and 129Xe NMR techniques. Anion size has the greatest effect on the spatial arrangement of the ILs and their mixtures in terms of the size of the non-polar domains and inter-ion distances. It was found that differences in coulombic attraction between oppositely charged ions arising from the distribution of charge density amongst the atoms of the anion also significantly influences these inter-ion distances. PALS and 129Xe NMR results pertaining to the free volume of these mixtures were found to strongly correlate with each other despite the vastly different timescales of these techniques. Furthermore, the excess free volumes calculated from each of these measurements were in excellent agreement with the excess volumes of mixing measured for the IL mixtures investigated. The correspondence of these techniques indicates that the static and dynamic free volume of these liquid mixtures are strongly linked. Consequently, fluxional processes such as hydrogen bonding do not significantly contribute to the free volumes of these liquids compared to the spatial arrangement of ions arising from their size, shape and coulombic attraction. Given the relationship between free volume and transport properties such as viscosity and conductivity, these results provide a link between the s
Trantidou T, Friddin M, Elani Y, et al., 2017, Engineering compartmentalized biomimetic micro- and nanocontainers, ACS Nano, Vol: 11, Pages: 6549-6565, ISSN: 1936-086X
Compartmentalization of biological content and function is a key architectural feature in biology, where membrane bound micro- and nanocompartments are used for performing a host of highly specialized and tightly regulated biological functions. The benefit of compartmentalization as a design principle is behind its ubiquity in cells and has led to it being a central engineering theme in construction of artificial cell-like systems. In this review, we discuss the attractions of designing compartmentalized membrane-bound constructs and review a range of biomimetic membrane architectures that span length scales, focusing on lipid-based structures but also addressing polymer-based and hybrid approaches. These include nested vesicles, multicompartment vesicles, large-scale vesicle networks, as well as droplet interface bilayers, and double-emulsion multiphase systems (multisomes). We outline key examples of how such structures have been functionalized with biological and synthetic machinery, for example, to manufacture and deliver drugs and metabolic compounds, to replicate intracellular signaling cascades, and to demonstrate collective behaviors as minimal tissue constructs. Particular emphasis is placed on the applications of these architectures and the state-of-the-art microfluidic engineering required to fabricate, functionalize, and precisely assemble them. Finally, we outline the future directions of these technologies and highlight how they could be applied to engineer the next generation of cell models, therapeutic agents, and microreactors, together with the diverse applications in the emerging field of bottom-up synthetic biology.
Many emerging technologies require materials with well-defined three-dimensional nanoscale architectures. Production of these structures is currently underpinned by self-assembling amphiphilic macromolecules or engineered all-DNA building blocks. Both of these approaches produce restricted ranges of crystal geometries due to synthetic amphiphiles' simple shape and limited specificity, or the technical difficulties in designing space-filling DNA motifs with targeted shapes. We have overcome these limitations with amphiphilic DNA nanostructures, or "C-Stars", that combine the design freedom and facile functionalization of DNA-based materials with robust hydrophobic interactions. C-Stars self-assemble into single crystals exceeding 40 μm in size with lattice parameters exceeding 20 nm.
Koch M, Wright KE, Otto O, et al., 2017, Plasmodium falciparum erythrocyte-binding antigen 175 triggers a biophysical change in the red blood cell that facilitates invasion., Proc Natl Acad Sci U S A, Vol: 114, Pages: 4225-4230
Invasion of the red blood cell (RBC) by the Plasmodium parasite defines the start of malaria disease pathogenesis. To date, experimental investigations into invasion have focused predominantly on the role of parasite adhesins or signaling pathways and the identity of binding receptors on the red cell surface. A potential role for signaling pathways within the erythrocyte, which might alter red cell biophysical properties to facilitate invasion, has largely been ignored. The parasite erythrocyte-binding antigen 175 (EBA175), a protein required for entry in most parasite strains, plays a key role by binding to glycophorin A (GPA) on the red cell surface, although the function of this binding interaction is unknown. Here, using real-time deformability cytometry and flicker spectroscopy to define biophysical properties of the erythrocyte, we show that EBA175 binding to GPA leads to an increase in the cytoskeletal tension of the red cell and a reduction in the bending modulus of the cell's membrane. We isolate the changes in the cytoskeleton and membrane and show that reduction in the bending modulus is directly correlated with parasite invasion efficiency. These data strongly imply that the malaria parasite primes the erythrocyte surface through its binding antigens, altering the biophysical nature of the target cell and thus reducing a critical energy barrier to invasion. This finding would constitute a major change in our concept of malaria parasite invasion, suggesting it is, in fact, a balance between parasite and host cell physical forces working together to facilitate entry.
Miller RM, Ces O, Brooks NJ, et al., 2017, Crystallization of Sodium Dodecyl Sulfate-Water Micellar Solutions under Linear Cooling, CRYSTAL GROWTH & DESIGN, Vol: 17, Pages: 2428-2437, ISSN: 1528-7483
The crystallization kinetics of sodium dodecyl sulfate (SDS)-water micellar solutions, under linear cooling conditions, were experimentally investigated using optical microscopy, differential scanning calorimetry, and infrared spectroscopy. Cooling rates were systematically varied, from 0.1 to 50 °C min–1, encompassing environmental to near-“isothermal” temperature changes, between 22 and −5 °C, for a reference concentration of 20% SDS-H2O. The cooling rate was shown to determine the dominant crystal morphologies, with platelets and needles predominating at the lowest and highest rates, respectively. The results were rationalized in terms of isothermal crystallization data and the time–temperature cooling profile. Rates 0.1, 5.0, and 10 °C min–1 yield morphologies and kinetics analogous to those of isothermal quenches at the corresponding crystallization temperature window. Nontrivial deviations were observed for intermediate rates (0.5, 1.0 °C min–1), due to commensurate changes in temperature and crystallization mechanism, accompanied by solute depletion. The polythermal metastable zone width was estimated, and the non-isothermal nucleation described by the Nývlt equation, while the Avrami and Kissinger models described overall crystallization kinetics. Our measurements quantify the impact of temperature gradients in the crystallization of ubiquitous SDS micellar solutions, for a range of practically relevant profiles incurred during manufacturing and storage.
Barlow NE, Smpokou E, Friddin MS, et al., 2017, Engineering plant membranes using droplet interface bilayers, Biomicrofluidics, Vol: 11, ISSN: 1932-1058
Droplet interface bilayers (DIBs) have become widely recognised as a robust platform for constructing model membranes and are emerging as a key technology for the bottom-up assembly of synthetic cell-like and tissue-like structures. DIBs are formed when lipid-monolayer coated water droplets are brought together inside a well of oil, which is excluded from the interface as the DIB forms. The unique features of the system, compared to traditional approaches (e.g., supported lipid bilayers, black lipid membranes, and liposomes), is the ability to engineer multi-layered bilayer networks by connecting multiple droplets together in 3D, and the capability to impart bilayer asymmetry freely within these droplet architectures by supplying droplets with different lipids. Yet despite these achievements, one potential limitation of the technology is that DIBs formed from biologically relevant components have not been well studied. This could limit the reach of the platform to biological systems where bilayer composition and asymmetry are understood to play a key role. Herein, we address this issue by reporting the assembly of asymmetric DIBs designed to replicate the plasma membrane compositions of three different plant species; Arabidopsis thaliana, tobacco, and oats, by engineering vesicles with different amounts of plant phospholipids, sterols and cerebrosides for the first time. We show that vesicles made from our plant lipid formulations are stable and can be used to assemble asymmetric plant DIBs. We verify this using a bilayer permeation assay, from which we extract values for absolute effective bilayer permeation and bilayer stability. Our results confirm that stable DIBs can be assembled from our plant membrane mimics and could lead to new approaches for assembling model systems to study membrane translocation and to screen new agrochemicals in plants.
McCarthy NL, Brooks NJ, Tyler AII, et al., 2017, A combined X-ray scattering and simulation study of halothane in membranes at raised pressures, Chemical Physics Letters, Vol: 671, Pages: 21-27, ISSN: 0009-2614
Using a combination of high pressure wide angle X-ray scattering exper-iments and molecular dynamics simulations, we probe the e ect of thearchetypal general anaesthetic halothane on the lipid hydrocarbon chainpacking and ordering in model bilayers and the variation in these parame-ters with pressure. Incorporation of halothane into the membrane causes anexpansion of the lipid hydrocarbon chain packing at all pressures but thee ect of halothane incorporation on the hydrocarbon chain order parameteris signi cantly reduced at elevated pressure.
We present a simple method for the multiplexed formation ofdroplet interface bilayers (DIBs) using a mechanically operatedlinear acrylic chamber array. To demonstrate the functionality ofthe chip design, a lipid membrane permeability assay is performed.We show that multiple, symmetric DIBs can be created andseparated using this robust low-cost approach.
Furse S, Brooks NJ, Woscholski R, et al., 2016, Pressure-dependent inverse bicontinuous cubic phase formation in a phosphatidylinositol 4-phosphate/phosphatidylcholine system, Chemical Data Collections, Vol: 3-4, Pages: 15-20, ISSN: 2405-8300
In this paper, we report the inositide-driven formation of an inverse bicontinuous cubic phase with space group Ia3d (QIIG, gyroid phase). The system under study consisted of distearoylphosphatidylinositol 4-phosphate (DSPIP) and dioleoylphosphatidylcholine at a molar ratio of 1:49, with a physiological concentration of magnesium ions at pH 7·4. The behaviour of the system was monitored as a function of temperature and pressure. The formation of the phase with Ia3d geometry was recorded repeatably at high pressure, and occurred more readily at higher temperatures. We conclude that the Ia3d phase formed is a thermodynamically stable structure, and that DSPIP is a potent source of membrane curvature that can drive the formation of mesophases with both 2- and 3D geometry.
Chan CL, Bolognesi G, Bhandarkar A, et al., 2016, DROPLAY: laser writing of functional patterns within biological microdroplet displays, Lab on a Chip, Vol: 16, Pages: 4621-4627, ISSN: 1473-0197
In this study, we introduce an optofluidic method for the rapid construction of large-area cell-sized droplet assemblieswith user-defined re-writable two-dimensional patterns of functional droplets. Light responsive water-in-oil dropletscapable of releasing fluorescent dye molecules upon exposure were generated and self-assembled into arrays in amicrofluidic device. This biological architecture was exploited by the scanning laser of a confocal microscope to ‘write’ userdefined patterns of differentiated (fluorescent) droplets in a network of originally undifferentiated (non-fluorescent)droplets. As a result, long lasting images were produced on a droplet fabric with droplets acting as pixels of a biologicalmonitor, which can be erased and re-written on-demand. Regio-specific light-induced droplet differentiation within a largepopulation of droplets provides a new paradigm for the rapid construction of bio-synthetic systems with potential as tissuemimics and biological display materials.
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