I22-Small angle scattering & Diffraction
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Diamond Proposal Number(s):
[21776]
Open Access
Abstract: The persulfate-initiated aqueous emulsion polymerization of 2,2,2-trifluoroethyl methacrylate (TFEMA) is studied by time-resolved small-angle X-ray scattering (SAXS) at 60 °C using a stirrable reaction cell. TFEMA was preferred to styrene because it offers much greater X-ray scattering contrast relative to water, which is essential for sufficient temporal resolution. The evolution in particle size is monitored by both in situ SAXS and ex situ DLS in the absence or presence of an anionic surfactant (sodium dodecyl sulfate, SDS). Post-mortem SAXS studies confirmed the formation of well-defined spherical latexes, with volume-average diameters of 353 ± 9 nm and 68 ± 4 nm being obtained for the surfactant-free and SDS formulations, respectively. 1H NMR spectroscopy studies of the equivalent laboratory-scale formulations indicated TFEMA conversions of 99% within 80 min and 93% within 60 min for the surfactant-free and SDS formulations, respectively. Comparable polymerization kinetics are observed for the in situ SAXS experiments and the laboratory-scale syntheses, with nucleation occurring after approximately 6 min in each case. After nucleation, scattering patterns are fitted using a hard sphere scattering model to determine the evolution in particle growth for both formulations. Moreover, in situ SAXS enables identification of the three main intervals (I, II, and III) that are observed during aqueous emulsion polymerization in the presence of surfactant. These intervals are consistent with those indicated by solution conductivity and optical microscopy studies. Significant differences between the surfactant-free and SDS formulations are observed, providing useful insights into the mechanism of emulsion polymerization.
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Jan 2021
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B21-High Throughput SAXS
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Diamond Proposal Number(s):
[21035]
Open Access
Abstract: The object of this study is a comparison between solid lipid nanoparticles and ethosomes for caffeic acid delivery through the skin. Caffeic acid is a potent antioxidant molecule whose cutaneous administration is hampered by its low solubility and scarce stability. In order to improve its therapeutic potential, caffeic acid has been encapsulated within solid lipid nanoparticles and ethosomes. The effect of lipid matrix has been evaluated on the morphology and size distribution of solid lipid nanoparticles and ethosomes loaded with caffeic acid. Particularly, morphology has been investigated by cryogenic transmission electron microscopy and small angle X-ray scattering, while mean diameters have been evaluated by photon correlation spectroscopy. The antioxidant power has been evaluated by the 2,2-diphenyl-1-picrylhydrazyl methodology. The influence of the type of nanoparticulate system on caffeic acid diffusion has been evaluated by Franz cells associated to the nylon membrane, while to evaluate caffeic acid permeation through the skin, an amperometric study has been conducted, which was based on a porcine skin-covered oxygen electrode. This apparatus allows measuring the O2 concentration changes in the membrane induced by polyphenols and H2O2 reaction in the skin. The antioxidative reactions in the skin induced by caffeic acid administered by solid lipid nanoparticles or ethosomes have been evaluated. Franz cell results indicated that caffeic acid diffusion from ethosomes was 18-fold slower with respect to solid lipid nanoparticles. The amperometric method evidenced the transdermal delivery effect of ethosome, indicating an intense antioxidant activity of caffeic acid and a very low response in the case of SLN. Finally, an irritation patch test conducted on 20 human volunteers demonstrated that both ethosomes and solid lipid nanoparticles can be safely applied on the skin.
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Jan 2021
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I19-Small Molecule Single Crystal Diffraction
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Sérgio M. F.
Vilela
,
Jorge A. R.
Navarro
,
Paula
Barbosa
,
Ricardo F.
Mendes
,
Germán
Pérez-sánchez
,
Harriott
Nowell
,
Duarte
Ananias
,
Filipe
Figueiredo
,
José R. B.
Gomes
,
João P. C.
Tomé
,
Filipe A.
Almeida Paz
Abstract: Porous robust materials are typically the primary selection of several industrial processes. Many of these compounds are, however, not robust enough to be used as multifunctional materials. This is typically the case of Metal–Organic Frameworks (MOFs) which rarely combine several different excellent functionalities into the same material. In this report we describe the simple acid–base postsynthetic modification of isotypical porous rare-earth-phosphonate MOFs into a truly multifunctional system, maintaining the original porosity features: [Ln(H3pptd)]·xSolvent [where Ln3+ = Y3+ (1) and (Y0.95Eu0.05)3+ (1_Eu)] are converted into [K3Ln(pptd)]·zSolvent [where Ln3+ = Y3+ (1K) and (Y0.95Eu0.05)3+ (1K_Eu)] by immersing the powder of 1 and 1_Eu into an ethanolic solution of KOH for 48 h. The K+-exchanged Eu3+-based material exhibits a considerable boost in CO2 adsorption, capable of being reused for several consecutive cycles. It can further separate C2H2 from CO2 from a complex ternary gas mixture composed of CH4, CO2, and C2H2. This high adsorption selectivity is, additionally, observed for other gaseous mixtures, such as C3H6 and C3H8, with all these results being supported by detailed theoretical calculations. The incorporation of K+ ions notably increases the electrical conductivity by 4 orders of magnitude in high relative humidity conditions. The conductivity is assumed to be predominantly protonic in nature, rendering this material as one of the best conducting MOFs reported to date.
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Jan 2021
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I22-Small angle scattering & Diffraction
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Diamond Proposal Number(s):
[15478, 14948, 12950]
Open Access
Abstract: Graphene oxide (GO) forms a well-aligned lyotropic liquid crystal (LC) phase in aqueous dispersions at relatively low concentrations. Under a remarkably wide range of shear rates, we report hitherto unobserved shear-induced polarized light image patterns, a Maltese cross combined with shear banding, recorded in real time and in situ during rheological measurements. This is shown to be a result of elastic flow instabilities that manifest as a helical flow in alternating bands of left- and right-handed helices, arising from a combination of shear flow and Taylor-type vortex flow. The instability is observed for LCs formed from large aspect ratio GO particles owing to their unique viscoelastic properties, but not for smaller aspect ratio particles. This phenomenon coincides with rheopecty and anomalous small-angle X-ray scattering patterns under shear flow, which confirm the instabilities. The results presented here could lead to advanced control over macroscopic periodic alignment in technologically relevant dispersions of two-dimensional material particles.
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Jan 2021
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Krios I-Titan Krios I at Diamond
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Diamond Proposal Number(s):
[14769]
Open Access
Abstract: The Tuberous Sclerosis Complex (TSC) protein complex (TSCC), comprising TSC1, TSC2, and TBC1D7, is widely recognised as a key integration hub for cell growth and intracellular stress signals upstream of the mammalian target of rapamycin complex 1 (mTORC1). The TSCC negatively regulates mTORC1 by acting as a GTPase-activating protein (GAP) towards the small GTPase Rheb. Both human TSC1 and TSC2 are important tumour suppressors, and mutations in them underlie the disease tuberous sclerosis.
We used single-particle cryo-EM to reveal the organisation and architecture of the complete human TSCC. We show that TSCC forms an elongated scorpion-like structure, consisting of a central “body”, with a “pincer” and a “tail” at the respective ends. The “body” is composed of a flexible TSC2 HEAT repeat dimer, along the surface of which runs the TSC1 coiled-coil backbone, breaking the symmetry of the dimer. Each end of the body is structurally distinct, representing the N- and C-termini of TSC1; a “pincer” is formed by the highly flexible N-terminal TSC1 core domains and a barbed “tail” makes up the TSC1 coiled-coil-TBC1D7 junction. The TSC2 GAP domain is found abutting the centre of the body on each side of the dimerisation interface, poised to bind a pair of Rheb molecules at a similar separation to the pair in activated mTORC1.
Our architectural dissection reveals the mode of association and topology of the complex, casts light on the recruitment of Rheb to the TSCC, and also hints at functional higher order oligomerisation, which has previously been predicted to be important for Rheb-signalling suppression.
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Jan 2021
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I13-2-Diamond Manchester Imaging
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Open Access
Abstract: Fetal immobilization affects skeletal development and can lead to severe malformations. Still, how mechanical load affects embryonic bone formation is not fully elucidated. This study combines mechanobiology, image analysis and developmental biology, to investigate the structural effects of muscular loading on embryonic long bones. We present a novel approach involving a semi-automatic workflow, to study the spatial and temporal evolutions of both hard and soft tissues in 3D without any contrast agent at micrometrical resolution. Using high-resolution phase-contrast-enhanced X-ray synchrotron microtomography, we compare the humeri of Splotch-delayed embryonic mice lacking skeletal muscles with healthy littermates. The effects of skeletal muscles on bone formation was studied from the first stages of mineral deposition (Theiler Stages 23 and 24) to just before birth (Theiler Stage 27). The results show that muscle activity affects both growth plate and mineralized regions, especially during early embryonic development. When skeletal muscles were absent, there was reduced mineralization, altered tuberosity size and location, and, at early embryonic stages, decreased chondrocyte density, size and elongation compared to littermate controls. The proposed workflow enhances our understanding of mechanobiology of early bone formation and could be implemented for the study of other complex biological tissues.
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Jan 2021
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B21-High Throughput SAXS
I03-Macromolecular Crystallography
I24-Microfocus Macromolecular Crystallography
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Tomasz
Uchański
,
Simonas
Masiulis
,
Baptiste
Fischer
,
Valentina
Kalichuk
,
Uriel
López-sánchez
,
Eleftherios
Zarkadas
,
Miriam
Weckener
,
Andrija
Sente
,
Philip
Ward
,
Alexandre
Wohlkonig
,
Thomas
Zogg
,
Han
Remaut
,
James
Naismith
,
Hugues
Nury
,
Wim
Vranken
,
A. Radu
Aricescu
,
Els
Pardon
,
Jan
Steyaert
Abstract: Nanobodies are popular and versatile tools for structural biology. They have a compact single immunoglobulin domain organization, bind target proteins with high affinities while reducing their conformational heterogeneity and stabilize multi-protein complexes. Here we demonstrate that engineered nanobodies can also help overcome two major obstacles that limit the resolution of single-particle cryo-electron microscopy reconstructions: particle size and preferential orientation at the water–air interfaces. We have developed and characterized constructs, termed megabodies, by grafting nanobodies onto selected protein scaffolds to increase their molecular weight while retaining the full antigen-binding specificity and affinity. We show that the megabody design principles are applicable to different scaffold proteins and recognition domains of compatible geometries and are amenable for efficient selection from yeast display libraries. Moreover, we demonstrate that megabodies can be used to obtain three-dimensional reconstructions for membrane proteins that suffer from severe preferential orientation or are otherwise too small to allow accurate particle alignment.
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Jan 2021
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I03-Macromolecular Crystallography
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Christopher M.
Furze
,
Ignacio
Delso
,
Enriqueta
Casal
,
Collette S.
Guy
,
Chloe
Seddon
,
Chelsea M.
Brown
,
Hadyn L.
Parker
,
Anjana
Radhakrishnan
,
Raul
Pacheco-gomez
,
Phillip J.
Stansfeld
,
Jesus
Angulo
,
Alexander D.
Cameron
,
Elizabeth
Fullam
Diamond Proposal Number(s):
[19880]
Open Access
Abstract: The Mycobacterium tuberculosis (Mtb) LpqY-SugABC ATP-binding cassette transporter is a recycling system that imports trehalose released during remodelling of the Mtb cell-envelope. As this process is essential for the virulence of the Mtb pathogen it may represent an important target for tuberculosis drug and diagnostic development, but the transporter specificity and molecular determinants of substrate recognition are unknown. To address this, we have determined the structural and biochemical basis of how mycobacteria transport trehalose using a combination of crystallography, STD NMR, molecular dynamics, site-directed mutagenesis, biochemical/biophysical assays and the synthesis of trehalose analogues. This analysis pinpoints key residues of the LpqY substrate binding lipoprotein that dictate substrate-specific recognition and has revealed which disaccharide modifications are tolerated. These findings provide critical insights into how the essential Mtb LpqY-SugABC transporter reuses trehalose and modified analogues, and specifies a framework that can be exploited for the design of new anti-tubercular agents and/or diagnostic tools.
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Jan 2021
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I10-Beamline for Advanced Dichroism
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Diamond Proposal Number(s):
[11784, 12943, 12958]
Open Access
Abstract: From the perspective of surface science, only the topmost atomic layers usually exhibit physical properties that are different to those of the bulk material, whereas the deeper layers are assumed to be bulk-like and remain largely unexplored. Going beyond conventional diffraction and imaging techniques, we have determined the depth dependence of the full 3D spin structure of magnetic skyrmions below the surface of a bulk Cu2OSeO3 sample using the polarization dependence of resonant elastic x-ray scattering (REXS). While the bulk spin configuration showed the anticipated Bloch type structure, it was found that the skyrmion lattice changes to a Néel twisting (i.e., with a different helicity angle) at the surface within a distance of several hundred nm. The exact surface helicity angle and penetration length of this twist have been determined, revealing the detailed internal structure of the skyrmion tube. It was found that the experimental penetration length of the Néel twisting is 7× longer than the theoretical value given by the ratio of J/D. This indicates that apart from the considered spin interactions, i.e., the Heisenberg exchange interaction J and the Dzyaloshinskii-Moriya interaction D, as well as the Zeeman interaction, other effects must play an important role. The findings suggest that the surface reconstruction of the skyrmion lattice is a universal phenomenon, stemming from the breaking of translational symmetry at the interface.
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Jan 2021
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I10-Beamline for Advanced Dichroism
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Diamond Proposal Number(s):
[20182]
Open Access
Abstract: A chiral bobber is a localized three-dimensional magnetization configuration, terminated by a singularity. Chiral bobbers coexist with magnetic skyrmions in chiral magnets, lending themselves to new types of skyrmion-complementary bits of information. However, the on-demand creation of bobbers, as well as their direct observation remained elusive. Here, we introduce a new mechanism for creating a stable chiral bobber lattice state via the proximity of two skyrmion species with comparable size. This effect is experimentally demonstrated in a
Cu
2
OSeO
3
/
[
Ta
/
CoFeB
/
MgO
]
4
heterostructure in which an exotic bobber lattice state emerges in the phase diagram of
Cu
2
OSeO
3
. To unambiguously reveal the existence of the chiral bobber lattice state, we have developed a novel characterization technique, magnetic truncation rod analysis, which is based on resonant elastic x-ray scattering.
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Jan 2021
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