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Recent reports from UN International Resource Panel call for double decoupling – decoupling of material use and related environmental impacts from economic growth. The construction and use of buildings constitute a significant portion of energy use, GHG emissions and extraction of materials in Europe. One of the central strategies addressing double decoupling in the construction industry is reuse and recycling of building materials and components that would limit raw material extraction and embodied emissions related to new products. A recent IPCC report states the urgency of limiting GHG emissions already by 2030 to remain below the 1.5-degree target. Given this timeframe, reducing emissions in the early stages of a buildings lifetime appears worth considering, meaning prioritizing embodied over operational emissions. The case study used for this article is an ongoing building project “Selbukassa” (“the Selbu box”) situated in Svartlamon – an experimental neighbourhood for urban ecology in Trondheim, Norway. It is a bottom-up, self-build project with a strong focus on reuse of materials. The 300 m2 building will house 4 families and is built reusing an old log house, as well as CLT-elements from a former pavilion used for exhibiting an art piece by the local artist Killi Olsen. Other salvaged materials and components include windows, doors, roof slate and most of the internal and external finishes. Due to the experimental status of the area and to allow for reuse of building components with lower energy performance, the fulfilment of energy requirements in the building code TEK 17 was not required by the local building authorities. The article examines the embodied and operational emissions of Selbukassa, and compares it with a reference building with no reused components and complying with energy requirements in the building code TEK17. Simien v.6.009 is used for energy simulations of the two cases, and LCA tool One Click LCA Norge for NS 3720:2018 with data sourced from Norwegian and other European EPDs is used for estimating embodied emissions. The article investigates the possible savings in emissions linked to reuse of materials, as well as the trade-offs between reused components with worse energy performance and the consequential higher energy demand. The article contributes to the discussion about the various implications of reuse of materials and more generally on how the built environment could respond more optimally to a combined emission and resource use reduction challenge.

With upgrades planned at several x-ray light sources, including improvements to beam quality and brilliance at high energies (> 20 keV), corresponding advances in area detector technology are needed. Pixel array detectors (PADs) are one class of detectors that can meet these needs. PADs feature highly customizable CMOS circuitry for signal processing and data streaming, which facilitates high frame rates and the exploration of various dynamic range extension techniques. The custom CMOS readout chip is mated at a pixel level to a sensor layer that may be chosen independently, providing a testbed for exploring sensors beyond silicon. Here, the development of a fast-framing, wide-dynamic-range PAD intended specifically for use at x-ray energies > 20 keV will be described.

The effect of introducing internal cavities on protein native structure and global stability has been well documented, but the consequences of these packing defects on folding free-energy landscapes have received less attention. We investigated the effects of cavity creation on the folding landscape of the leucine-rich repeat protein pp32 by high-pressure (HP) and urea-dependent NMR and high-pressure small-angle X-ray scattering (HPSAXS). Despite a modest global energetic perturbation, cavity creation in the N-terminal capping motif (N-cap) resulted in very strong deviation from two-state unfolding behavior. In contrast, introduction of a cavity in the most stable, C-terminal half of pp32 led to highly concerted unfolding, presumably because the decrease in stability by the mutations attenuated the N- to C-terminal stability gradient present in WT pp32. Interestingly, enlarging the central cavity of the protein led to the population under pressure of a distinct intermediate in which the N-cap and repeats 1–4 were nearly completely unfolded, while the fifth repeat and the C-terminal capping motif remained fully folded. Thus, despite modest effects on global stability, introducing internal cavities can have starkly distinct repercussions on the conformational landscape of a protein, depending on their structural and energetic context.

A high-speed pixel array detector for time-resolved X-ray imaging at synchrotrons has been developed. The ability to isolate single synchrotron bunches makes it ideal for time-resolved dynamical studies., A wide-dynamic-range imaging X-ray detector designed for recording successive frames at rates up to 10 MHz is described. X-ray imaging with frame rates of up to 6.5 MHz have been experimentally verified. The pixel design allows for up to 8–12 frames to be stored internally at high speed before readout, which occurs at a 1 kHz frame rate. An additional mode of operation allows the integration capacitors to be re-addressed repeatedly before readout which can enhance the signal-to-noise ratio of cyclical processes. This detector, along with modern storage ring sources which provide short (10–100 ps) and intense X-ray pulses at megahertz rates, opens new avenues for the study of rapid structural changes in materials. The detector consists of hybridized modules, each of which is comprised of a 500 µm-thick silicon X-ray sensor solder bump-bonded, pixel by pixel, to an application-specific integrated circuit. The format of each module is 128 × 128 pixels with a pixel pitch of 150 µm. In the prototype detector described here, the three-side buttable modules are tiled in a 3 × 2 array with a full format of 256 × 384 pixels. The characteristics, operation, testing and application of the detector are detailed.

Structuring ionic solids at the nanoscale with block copolymers (BCPs) is notoriously difficult due to solvent incompatibilities and strong driving forces for crystallization of the inorganic material. Here, we demonstrate that elucidating pathway complexity in the BCP-directed self-assembly of an ionic solid, amorphous calcium phosphate (ACP), is a key component in obtaining nanostructured, bulk composite materials in which the nanostructure is the result of thermodynamically controlled BCP self-assembly, i.e., exhibiting sequences of bulk morphologies as known from typical equilibrium BCP phase diagrams. Specifically, we identify three critical pathway “decision points” for the evaporation-induced self-assembly of composites from ultrasmall, organosilicate-modified amorphous calcium phosphate nanoparticles (osm-ACP-NPs) and poly(isoprene)-block-poly(2-(dimethylamino)ethyl methacrylate) (PI-b-PDMAEMA) block copolymers. Using this strategy enabled us to obtain composites with hexagonal, cubic network, and...

Using the EMC algorithm, the three-dimensional intensity was successfully reconstructed from millions of non-oriented, sparse data frames collected from a hen egg-white lysozyme crystal rotating about a single axis. The protein structure was solved from the reconstructed intensity. This result is encouraging for the development of synchrotron-based serial microcrystallography., X-ray free-electron lasers (XFELs) have inspired the development of serial femtosecond crystallography (SFX) as a method to solve the structure of proteins. SFX datasets are collected from a sequence of protein microcrystals injected across ultrashort X-ray pulses. The idea behind SFX is that diffraction from the intense, ultrashort X-ray pulses leaves the crystal before the crystal is obliterated by the effects of the X-ray pulse. The success of SFX at XFELs has catalyzed interest in analogous experiments at synchrotron-radiation (SR) sources, where data are collected from many small crystals and the ultrashort pulses are replaced by exposure times that are kept short enough to avoid significant crystal damage. The diffraction signal from each short exposure is so ‘sparse’ in recorded photons that the process of recording the crystal intensity is itself a reconstruction problem. Using the EMC algorithm, a successful reconstruction is demonstrated here in a sparsity regime where there are no Bragg peaks that conventionally would serve to determine the orientation of the crystal in each exposure. In this proof-of-principle experiment, a hen egg-white lysozyme (HEWL) crystal rotating about a single axis was illuminated by an X-ray beam from an X-ray generator to simulate the diffraction patterns of microcrystals from synchrotron radiation. Millions of these sparse frames, typically containing only ∼200 photons per frame, were recorded using a fast-framing detector. It is shown that reconstruction of three-dimensional diffraction intensity is possible using the EMC algorithm, even with these extremely sparse frames and without knowledge of the rotation angle. Further, the reconstructed intensity can be phased and refined to solve the protein structure using traditional crystallographic software. This suggests that synchrotron-based serial crystallography of micrometre-sized crystals can be practical with the aid of the EMC algorithm even in cases where the data are sparse.

Stimuli-responsive materials have attracted great interest in catalysis, sensing, and drug delivery applications and are typically constituted by soft components. We present a one-pot synthetic method for a type of inorganic silica-based shape change material that is responsive to water vapor exposure. After the wetting treatment, the cross-sectional shape of aminated mesoporous silica nanoparticles (MSNs) with hexagonal pore lattice changed from hexagonal to six-angle-star, accompanied by the loss of periodic mesostructural order. Nitrogen sorption measurements suggested that the wetting treatment induced a shrinkage of mesopores resulting in a broad size distribution and decreased mesopore volume. Solid-state (29)Si nuclear magnetic resonance (NMR) spectroscopy of samples after wetting treatment displayed a higher degree of silica condensation, indicating that the shape change was associated with the formation of more siloxane bonds within the silica matrix. On the basis of material characterization results, a mechanism for the observed anisotropic shrinkage is suggested based on a buckling deformation induced by capillary forces in the presence of a threshold amount of water vapor available beyond a humidity of about 50%. The work presented here may open a path toward novel stimuli-responsive materials based on inorganic components.

Analysis of fresh capillary blood using darkfield microscopy (FCB-DM) is a point of care screening tool, used by healthcare practitioners in Australia. However, the relationship between the outcomes of FCB-DM measures and the automated venous haematology measures has not been determined. Objectives The aim of this study was to compare differential white blood cell counts obtained using automated haematology and FCB-DM. Methods Data of 125 individuals were collected either retrospectively ( n = 74) or from participants specifically recruited for the project ( n = 51). Retrospective data were collected from active files at a naturopathic clinic. Newly recruited participants provided a fasting capillary blood sample for FCB-DM analysis within 1 h of providing a venous blood sample at a commercial laboratory for automated haematologic analysis. Results The mean score of neutrophils was found to be higher, and lymphocytes and basophils to be lower, in FCB-DM analysis ( p r = 0.60, p r = 0.63, p r s = 0.32, p r s = 0.596, p Conclusion Despite significant differences in the mean scores of cells counts, significant correlations between the data obtained by the two techniques for neutrophil, lymphocyte, monocyte and eosinophil cells were observed. Given the small amount of blood sample required, FCB-DM would have an advantage in clinical practice, though further research is required to determine the clinical implications of the FCB-DM cell counts.

Observation of theorized glass-to-liquid transitions between low-density amorphous (LDA) and high-density amorphous (HDA) water states had been stymied by rapid crystallization below the homogeneous water nucleation temperature (∼235 K at 0.1 MPa). We report optical and X-ray observations suggestive of glass-to-liquid transitions in these states. Crack healing, indicative of liquid, occurs when LDA ice transforms to cubic ice at 160 K, and when HDA ice transforms to the LDA state at temperatures as low as 120 K. X-ray diffraction study of the HDA to LDA transition clearly shows the characteristics of a first-order transition. Study of the glass-to-liquid transitions in nanoconfined aqueous solutions shows them to be independent of the solute concentrations, suggesting that they represent an intrinsic property of water. These findings support theories that LDA and HDA ice are thermodynamically distinct and that they are continuously connected to two different liquid states of water.

Next-generation synchrotron radiation sources, such as X-ray free-electron lasers, energy recovery linacs, and ultra-low-emittance storage rings, are catalyzing novel methods of biomolecular microcrystallography and solution scattering. These methods are described and future trends are predicted. Importantly, there is a growing realization that serial microcrystallography and certain cutting-edge solution scattering experiments can be performed at existing storage ring sources by utilizing new technology. In this sense, next-generation sources are serving two distinct functions, namely, provision of new capabilities that require the newer sources and inspiration of new methods that can be performed at existing sources.

This paper describes a solid-state image sensor for high-speed X-ray imaging. The sensor is made up of a light sensitive detector layer bump-bonded to a readout integrated circuit (ROIC). The detector layer is high resistivity n-type silicon and is fully depleted in operation. The p-implanted islands are used to define pixel regions with 100-μm × 100-μm area. The detector layer contains 852 × 209 pixels indium bump-bonded to four identical CMOS ROICs. Each ROIC contains 213 × 209 pixels and is fabricated using a 0.25-μm CMOS process. The ROIC utilizes a capacitive transimpedance amplifier-type front-end coupled to a switched capacitor analog memory. This architecture allows storage of eight frames for high-speed burst imaging of up to a million frames per second, followed by a slower multiplexed readout. Details of the sensor design and operation are presented together with characterization results.

Aberration-corrected optics have made electron microscopy at atomic resolution a widespread and often essential tool for characterizing nanoscale structures. Image resolution has traditionally been improved by increasing the numerical aperture of the lens (α) and the beam energy, with the state-of-the-art at 300 kiloelectronvolts just entering the deep sub-angstrom (that is, less than 0.5 angstrom) regime. Two-dimensional (2D) materials are imaged at lower beam energies to avoid displacement damage from large momenta transfers, limiting spatial resolution to about 1 angstrom. Here, by combining an electron microscope pixel-array detector with the dynamic range necessary to record the complete distribution of transmitted electrons and full-field ptychography to recover phase information from the full phase space, we increase the spatial resolution well beyond the traditional numerical-aperture-limited resolution. At a beam energy of 80 kiloelectronvolts, our ptychographic reconstruction improves the image contrast of single-atom defects in MoS2 substantially, reaching an information limit close to 5α, which corresponds to an Abbe diffraction-limited resolution of 0.39 angstrom, at the electron dose and imaging conditions for which conventional imaging methods reach only 0.98 angstrom.

Next-generation, atomically thin devices require in-plane, one-dimensional heterojunctions to electrically connect different two-dimensional (2D) materials. However, the lattice mismatch between most 2D materials leads to unavoidable strain, dislocations, or ripples, which can strongly affect their mechanical, optical, and electronic properties. We have developed an approach to map 2D heterojunction lattice and strain profiles with sub-picometer precision and to identify dislocations and out-of-plane ripples. We collected diffraction patterns from a focused electron beam for each real-space scan position with a high-speed, high dynamic range, momentum-resolved detector - the electron microscope pixel array detector (EMPAD). The resulting four-dimensional (4D) phase space datasets contain the full spatially resolved lattice information of the sample. By using this technique on tungsten disulfide (WS2) and tungsten diselenide (WSe2) lateral heterostructures, we have mapped lattice distortions with 0.3 pm precision across multi-micron fields of view and simultaneously observed the dislocations and ripples responsible for strain relaxation in 2D laterally-epitaxial structures.

A one-pot synthesis approach is described to generate ordered mesoporous crystalline γ-alumina–carbon composites and ordered mesoporous crystalline γ-alumina materials via the combination of soft and hard-templating chemistries using block copolymers as soft structure-directing agents. Periodically ordered alumina hybrid mesostructures were generated by self-assembly of a poly(isoprene)-block-poly(styrene)-block-poly(ethylene oxide) terpolymer, n-butanol and aluminum tri-sec-butoxide derived sols in organic solvents. The triblock terpolymer was converted into a rigid carbon framework during thermal annealing under nitrogen to support and preserve the ordered mesoporous crystalline γ-alumina–carbon composite structures up to 1200 °C. The carbon matrix was subsequently removed in a second heat treatment in air to obtain ordered mesoporous crystalline γ-alumina structures. Such thermally stable, highly crystalline, and periodically ordered mesoporous ceramic and ceramic–carbon composite materials may be promising candidates for various high temperature catalysis, separation, and energy-related applications.

Crystalline transition metal oxides with controlled mesopore architectures are in increasing demand to enhance the performance of energy conversion and storage devices. Solution based block copolymer self-assembly routes to achieve ordered mesoporous and crystalline titania have been studied for more than a decade, but have so far mostly been limited to water and alcohol dispersible polymers. This constraint has limited the accessible morphology space as well as structural dimensions. Moreover, synthetic approaches are mostly performed in a trial-and-error fashion using chemical intuition rather than being based on well-defined design parameters. We present solubility design guidelines that facilitate coassembly with highly amphiphilic block copolymers, enabling the formation of ordered structures with diverse length scales (d10 = 13.8–63.0 nm) and bulk-type morphologies. Thus, highly ordered and crystalline titania with the largest reported pores (d = 32.3 nm) was demonstrated for such a coassembly approach without the use of pore-expanders. Furthermore, the use of an ABC triblock terpolymer system led to a 3D ordered network morphology. In all cases, subsequent calcination treatments, such as the CASH procedure, enabled the formation of highly crystalline mesoporous materials while preserving the mesostructure.

The advantages of a novel wide dynamic range hard X-ray detector are demonstrated for (ptychographic) coherent X-ray diffractive imaging., Coherent (X-ray) diffractive imaging (CDI) is an increasingly popular form of X-ray microscopy, mainly due to its potential to produce high-resolution images and the lack of an objective lens between the sample and its corresponding imaging detector. One challenge, however, is that very high dynamic range diffraction data must be collected to produce both quantitative and high-resolution images. In this work, hard X-ray ptychographic coherent diffractive imaging has been performed at the P10 beamline of the PETRA III synchrotron to demonstrate the potential of a very wide dynamic range imaging X-ray detector (the Mixed-Mode Pixel Array Detector, or MM-PAD). The detector is capable of single photon detection, detecting fluxes exceeding 1 × 108 8-keV photons pixel−1 s−1, and framing at 1 kHz. A ptychographic reconstruction was performed using a peak focal intensity on the order of 1 × 1010 photons µm−2 s−1 within an area of approximately 325 nm × 603 nm. This was done without need of a beam stop and with a very modest attenuation, while ‘still’ images of the empty beam far-field intensity were recorded without any attenuation. The treatment of the detector frames and CDI methodology for reconstruction of non-sensitive detector regions, partially also extending the active detector area, are described.

Properties arising from ordered periodic mesostructures are often obscured by small, randomly oriented domains and grain boundaries. Bulk macroscopic single crystals with mesoscale periodicity are needed to establish fundamental structure-property correlations for materials ordered at this length scale (10-100 nm). A solvent-evaporation-induced crystallization method providing access to large (millimeter to centimeter) single-crystal mesostructures, specifically bicontinuous gyroids, in thick films (>100 µm) derived from block copolymers is reported. After in-depth crystallographic characterization of single-crystal block copolymer-preceramic nanocomposite films, the structures are converted into mesoporous ceramic monoliths, with retention of mesoscale crystallinity. When fractured, these monoliths display single-crystal-like cleavage along mesoscale facets. The method can prepare macroscopic bulk single crystals with other block copolymer systems, suggesting that the method is broadly applicable to block copolymer materials assembled by solvent evaporation. It is expected that such bulk single crystals will enable fundamental understanding and control of emergent mesostructure-based properties in block-copolymer-directed metal, semiconductor, and superconductor materials.

A novel strategy for fabrication of ordered ceramic–metal nanocomposites was demonstrated by multifunctional block copolymer/metal nanoparticle self-assembly. Hybrid organic–inorganic block copolymer poly(3-methacryloxypropyl-T8-heptaisobutyl-polyhedral oligomeric silsesquioxane-block-N,N-dimethylaminoethyl methacrylate) was synthesized and used as a bi-functional structure directing agent for ligand-stabilized platinum nanoparticles to form ordered organic–inorganic nanocomposites with dense loading of inorganic species in both microphase separated domains. Subsequently, thin films of the hybrid material were converted to ordered silica (ceramic)–platinum (metal) nanocomposites via UV-assisted ozonolysis. This is the first time ordered ceramic–metal nanocomposites were achieved through a bottom-up approach, opening up opportunities for the design and synthesis of a broad range of ordered inorganic–inorganic nanocomposites.

The overall signal-to-noise ratio per unit dose for X-ray diffraction data from protein crystals can be improved by reducing the mass and density of all material surrounding the crystals. This article demonstrates a path towards the practical ultimate in background reduction by use of atomically thin graphene sheets as a crystal mounting platform for protein crystals. The results show the potential for graphene in protein crystallography and other cases where X-ray scatter from the mounting material must be reduced and specimen dehydration prevented, such as in coherent X-ray diffraction imaging of microscopic objects.

Mesoporous nanomaterials have attracted widespread interest because of their structural versatility for applications including catalysis, separation, and nanomedicine. We report a one-pot synthesis method for a class of mesoporous silica nanoparticles (MSNs) containing both cubic and hexagonally structured compartments within one particle. These multicompartment MSNs (mc-MSNs) consist of a core with cage-like cubic mesoporous morphology and up to four branches with hexagonally packed cylindrical mesopores epitaxially growing out of the cubic core vertices. The extent of cylindrical mesostructure growth can be controlled via a single additive in the synthesis. Results suggest a path toward high levels of architectural complexity in locally amorphous, mesostructured nanoparticles, which could enable tuning of different pore environments of the same particle for specific chemistries in catalysis or drug delivery.

A proposed design for a reconfigurable x-ray Pixel Array Detector (PAD) is described. It operates by integrating a high-end commercial field programmable gate array (FPGA) into a 3-layer device along with a high-resistivity diode detection layer and a custom, application-specific integrated circuit (ASIC) layer. The ASIC layer contains an energy-discriminating photon-counting front end with photon hits streamed directly to the FPGA via a massively parallel, high-speed data connection. FPGA resources can be allocated to perform user defined tasks on the pixel data streams, including the implementation of a direct time autocorrelation function (ACF) with time resolution down to 100 ns. Using the FPGA at the front end to calculate the ACF reduces the required data transfer rate by several orders of magnitude when compared to a fast framing detector. The FPGA-ASIC high-speed interface, as well as the in-FPGA implementation of a real-time ACF for x-ray photon correlation spectroscopy experiments has been designed and simulated. A 16×16 pixel prototype of the ASIC has been fabricated and is being tested.

The so-called Red Salt Clay (T4) is deposited as clay-rich clastic sediment at the base of the Aller-series forming a persistent lateral layer of up to 20 m thickness above the lower Zechstein-series. The clay layers may act as a protective shield in the hanging wall of gas storages or underground repositories in salt formations, thus resulting in a multi-barrier system. As a proof of its reliability comprehensive hydro-mechanical investigations were performed on clay samples recovered at different sites in Germany. Most important, rock tightness against various fluids was confirmed in the lab and field-scale. Remarkably, only if the fluid pressure equalises the acting minimal stress (i.e. violence of the “minimum stress criterion”) a significant increase of permeability is observed (“pathway dilatation”) but no macro-frac. However, the material properties from different locations showed a significant variability according to different burial depths. Thus the Red Salt Clay may act as natural analogue, representing the material variability of various indurated clays. In addition, the existing knowledge gained from practical mining activities can be used to evaluate extreme in situ loading conditions.

A method for in situ X-ray reflectivity measurements on the millisecond time scale is described, and its use for measuring interdiffusion in metallic multilayers is illustrated., A technique for measuring interdiffusion in multilayer materials during rapid heating using X-ray reflectivity is described. In this technique the sample is bent to achieve a range of incident angles simultaneously, and the scattered intensity is recorded on a fast high-dynamic-range mixed-mode pixel array detector. Heating of the multilayer is achieved by electrical resistive heating of the silicon substrate, monitored by an infrared pyrometer. As an example, reflectivity data from Al/Ni heated at rates up to 200 K s−1 are presented. At short times the interdiffusion coefficient can be determined from the rate of decay of the reflectivity peaks, and it is shown that the activation energy for interdiffusion is consistent with a grain boundary diffusion mechanism. At longer times the simple analysis no longer applies because the evolution of the reflectivity pattern is complicated by other processes, such as nucleation and growth of intermetallic phases.

Several charge integrating CMOS pixel front-ends utilizing charge removal techniques have been fabricated to extend dynamic range for x-ray diffraction applications at synchrotron sources and x-ray free electron lasers (XFELs). The pixels described herein build on the Mixed Mode Pixel Array Detector (MM-PAD) framework, developed previously by our group to perform high dynamic range imaging. These new pixels boast several orders of magnitude improvement in maximum flux over the MM-PAD, which is capable of measuring a sustained flux in excess of 10$^{8}$ x-rays/pixel/second while maintaining sensitivity to smaller signals, down to single x-rays. To extend dynamic range, charge is removed from the integration node of the front-end amplifier without interrupting integration. The number of times this process occurs is recorded by a digital counter in the pixel. The parameter limiting full well is thereby shifted from the size of an integration capacitor to the depth of a digital counter. The result is similar to that achieved by counting pixel array detectors, but the integrators presented here are designed to tolerate a sustained flux >10$^{11}$ x-rays/pixel/second. Pixel front-end linearity was evaluated by direct current injection and results are presented. A small-scale readout ASIC utilizing these pixel architectures has been fabricated and the use of these architectures to increase single x-ray pulse dynamic range at XFELs is discussed briefly., 6 pages, 7 figures

Carbonic anhydrases are mostly zinc metalloenzymes that catalyze the reversible hydration/dehydration of CO2/HCO3 (-) Previously, the X-ray crystal structures of CO2-bound holo (zinc-bound) and apo (zinc-free) human carbonic anhydrase IIs (hCA IIs) were captured at high resolution. Here, we present sequential timeframe structures of holo- [T = 0 s (CO2-bound), 50 s, 3 min, 10 min, 25 min, and 1 h] and apo-hCA IIs [T = 0 s, 50 s, 3 min, and 10 min] during the "slow" release of CO2 Two active site waters, WDW (deep water) and WDW' (this study), replace the vacated space created on CO2 release, and another water, WI (intermediate water), is seen to translocate to the proton wire position W1. In addition, on the rim of the active site pocket, a water W2' (this study), in close proximity to residue His64 and W2, gradually exits the active site, whereas His64 concurrently rotates from pointing away ("out") to pointing toward ("in") active site rotameric conformation. This study provides for the first time, to our knowledge, structural "snapshots" of hCA II intermediate states during the formation of the His64-mediated proton wire that is induced as CO2 is released. Comparison of the holo- and apo-hCA II structures shows that the solvent network rearrangements require the presence of the zinc ion.

This work examines the effect of perturbing the position of bound CO(2) in the active site of human carbonic anhydrase II (HCA II) on catalysis. Variants of HCA II in which Val143 was replaced with hydrophobic residues Ile, Leu, and Ala were examined. The efficiency of catalysis in the hydration of CO(2) for these variants was characterized by (18)O exchange mass spectrometry, and their structures were determined by X-ray crystallography at 1.7-1.5 Å resolution. The most hydrophobic substitutions, V143I and V143L, showed decreases in the level of catalysis, as much as 20-fold, while the replacement by the smaller V143A mutation showed an only moderate 2-fold decrease in activity. Structural data for all three variants show no significant change in the overall position of amino acid side chains in the active site compared with the wild type. However, V143A HCA II showed additional ordered water molecules in the active site compared to the number for the wild type. To further investigate the decrease in the catalytic efficiency of V143I HCA II, an X-ray crystallographic CO(2) entrapment experiment was performed to 0.93 Å resolution. This structure revealed an unexpected shift in the CO(2) substrate toward the zinc-bound solvent, placing it ~0.3 Ǻ closer than previously observed in the wild type in conjunction with the observed dual occupancy of the product bicarbonate, presumably formed during the acquisition of data. These data suggest that the Ile substitution at position 143 reduced the catalytic efficiency, which is likely due to steric crowding resulting in destabilization of the transition state for conversion of CO(2) into bicarbonate and a decreased product dissociation rate.

We report the room temperature formation of aminated mesoporous silica nanoparticles (NH2-MSNs) by means of co-condensation of different molar ratios of tetraethyl orthosilicate (TEOS) and 3-aminopropyl triethoxysilane (APTES) in the synthesis feed. The resulting materials are characterized by a combination of transmission electron microscopy (TEM), small-angle X-ray scattering (SAXS), Fourier transform infrared (FTIR) spectroscopy, thermogravimetric analysis (TGA), and N2 adsorption/desorption measurements. Analysis reveals that an increase in APTES loading (mol %) leads to structural transitions in the MSNs from hexagonal (0–49 mol %) to cubic Pm3n (54–64 mol %) to disordered at very high APTES amounts (69 mol %). Investigation of structural evolution during cubic Pm3n particle synthesis reveals early particle formation stages that are surprisingly similar to those discussed in recent literature on nonclassical single crystal growth. These include significant heterogeneities in particle density despit...

Background In the cellular environment, macromolecules occupy about 30% of a cell's volume. In this crowded environment, proteins behave very differently than in dilute solution where scientists typically study the properties of proteins. For this reason, recent studies have investigated proteins in cell-like crowded conditions so as to understand if this changes their properties. The present study was performed to examine if molecular crowding impedes the protein unfolding process that is known to occur upon the application of high pressure. Methods Crowding of staphylococcal nuclease (SNase) was induced by dissolving low concentrations of SNase in high concentrations of crowding agents (16 wt.% or 25 wt.% PEG 3000 or 16 wt.% Dextran T10). SNase unfolding was then monitored via tryptophan fluorescence as pressure was applied. Results Fluorescence spectra can be decomposed into the sum of two components indicative, respectively, of native and unfolded states, and the center of spectral mass was then used as a measure of the degree of protein unfolding. It was found that SNase unfolding as a function of pressure was impeded in crowded solutions. These results suggest that crowded environments, such as those found in the cellular cytoplasm, may also impede high-pressure protein unfolding in cells. General significance This is the first report on the effect of crowding on the pressure-induced unfolding of a protein (staphylococcal nuclease) monitored via tryptophan fluorescence.

Diminished taste acuity (hypogeusia) has been linked to zinc deficiency in humans and animals. This phenomenon has been exploited in the Zinc Taste Test (ZTT), a taste acuity test commonly employed by Australian naturopaths. However, its validity has not yet been firmly established.A systematic search of several key databases was conducted. Only studies in which there were full reports of clinical trials comparing the ZTT to at least one other zinc test within the same sample population were included.Three (3) studies matched the criteria for inclusion. Study I compared the ZTT with sweat zinc in patients with food intolerance, reporting moderate correlation. Study II recruited pregnant women using the ZTT and serum zinc to assess zinc status, with above 70% congruence between the two tests at the start of the trial and 100% congruence at the end. Study III also recruited pregnant women at three stages during gestation, assessing ZTT and leukocyte zinc initially, later adding dietary zinc intake and at delivery cord blood zinc. No significant correlation was found between the results of these different methods; however, statistically significant differences in the ZTT responders (tasters and nontasters) were found for pregnancy outcomes.The methodology of the three studies is critically discussed. Although depletion of zinc leads to decreased taste acuity, it does not explain all cases of hypogeusia. Various other influences on taste perception are discussed in relation to the validity of the ZTT. Stringent exclusion criteria are therefore mandatory to increase specificity. Large variations from the original test design have been identified. The laboratory assays of zinc in these studies are also lacking sensitivity to accurately assess zinc status.To date, there are no tests that are both sensitive and specific that accurately assess marginal zinc status in humans. The ZTT, albeit widely used, does not fill this void, and further research is needed.

The production of high-performance carbon nanotube (CNT) materials demands understanding of the growth behavior of individual CNTs as well as collective effects among CNTs. We demonstrate the first use of grazing incidence small-angle X-ray scattering to monitor in real time the synthesis of CNT films by chemical vapor deposition. We use a custom-built cold-wall reactor along with a high-speed pixel array detector resulting in a time resolution of 10 msec. Quantitative models applied to time-resolved X-ray scattering patterns reveal that the Fe catalyst film first rapidly dewets into well-defined hemispherical particles during heating in a reducing atmosphere, and then the particles coarsen slowly upon continued annealing. After introduction of the carbon source, the initial CNT diameter distribution closely matches that of the catalyst particles. However, significant changes in CNT diameter can occur quickly during the subsequent CNT self-organization process. Correlation of time-resolved orientation data to X-ray scattering intensity and height kinetics suggests that the rate of self-organization is driven by both the CNT growth rate and density, and vertical CNT growth begins abruptly when CNT alignment reaches a critical threshold. The dynamics of CNT size evolution and self-organization vary according to the catalyst annealing conditions and substrate temperature. Knowledge of these intrinsically rapid processes is vital to improve control of CNT structure and to enable efficient manufacturing of high-density arrays of long, straight CNTs.

There is growing excitement in the synchrotron materials science community about the potential of nearly diffraction-limited, high-repetition rate, hard X-ray sources, such as an Energy Recovery Linac (ERL) or an Ultimate Storage Ring (USR), and that these sources will pave the way to scientific insights and discoveries not possible with existing facilities. These future sources will deliver highly coherent, nearly diffraction-limited X-ray beams that will power ultra-intense, nanometer-scale X-ray probes and imaging capabilities approaching atomic resolution. They will produce X-ray pulses at MHz to GHz repetition rates and span pulse durations from below 50 femtoseconds to tens of picoseconds, enabling new classes of experiments in hard X-ray science.

High-pressure methods for solving protein structures by X-ray crystallography and NMR are maturing. These techniques are beginning to impact our understanding of thermodynamic and structural features that define not only the protein's native conformation, but also the higher free energy conformations. The ability of high-pressure methods to visualize these mostly unexplored conformations provides new insight into protein function and dynamics. In this review, we begin with a historical discussion of high-pressure structural studies, with an eye toward early results that paved the way to mapping the multiple conformations of proteins. This is followed by an examination of several recent studies that emphasize different strengths and uses of high-pressure structural studies, ranging from basic thermodynamics to the suggestion of high-pressure structural methods as a tool for protein engineering.

Techniques are described for X-ray diffraction combining micrometer-scale spatial resolution with microsecond-scale temporal resolution for studying rapid localized irreversible transformations in materials., A pair of techniques have been developed for performing time-resolved X-ray microdiffraction on irreversible phase transformations. In one technique capillary optics are used to focus a high-flux broad-spectrum X-ray beam to a 60 µm spot size and a fast pixel array detector is used to achieve temporal resolution of 55 µs. In the second technique the X-rays are focused with Kirkpatrick–Baez mirrors to achieve a spatial resolution better than 10 µm and a fast shutter is used to provide temporal resolution better than 20 µs while recording the diffraction pattern on a (relatively slow) X-ray CCD camera. Example data from experiments are presented where these techniques are used to study self-propagating high-temperature synthesis reactions in metal laminate foils.

The aetiology of non-syndromic cleft lip and palate has as yet not been clearly defined. Familial relationships, environmental toxins and nutritional status have all been considered without conclusive results, although in some studies a potential link between non-syndromic cleft lip and palate and any one or more of these factors has been proposed. Elevated stress, particularly an extended term of traumatic stress, can lead to oxidative damage at the cellular level via hypothalamus-pituitary-adrenal (HPA) axis dysregulation, high cortisol and cytokine production. The effect of this hormonal shift is to re-direct the blood supply to the mother's muscles, thereby reducing the supply to the placenta, causing a potential nutritional deficiency which may then result in a genetic alteration in the foetus. Mothers with a child aged two years or younger who had been born with a cleft, who were members of CleftPals, a family support group, volunteered to be participants in this qualitative study. The research first called for a survey to be completed by the mother and this was then followed by an interview conducted by the researcher. The study involved families living in the three eastern States of Australia. The results suggest that physical and/or emotional stress may well be implicated in clefting. While little work has been done in considering stress as a causal factor, the existing literature suggests, as does this study, that elevated stress levels at, or soon after, conception appear to affect foetal development.

Long-standing evidence suggests that plasticity in metals may proceed in an intermittent fashion. While the documentation of intermittency in plastically deforming materials has been achieved in several experimental settings, efforts to draw connections from dislocation motion and structure development to stress relaxation have been limited, especially in the bulk of deforming polycrystals. This work uses high energy x-ray diffraction measurements to build these links by characterizing plastic deformation events inside individual deforming grains in both the titanium alloy, Ti-7Al, and the magnesium alloy, AZ31. This analysis is performed by combining macroscopic stress relaxation data, complete grain stress states found using far-field high energy diffraction microscopy, and rapid x-ray diffraction spot measurements made using a Mixed-Mode Pixel Array Detector. Changes in the dislocation content within the deforming grains are monitored using the evolution of the full 3-D shapes of the diffraction spot intensity distributions in reciprocal space. The results for the Ti-7Al alloy show the presence of large stress fluctuations in contrast to AZ31, which shows a lesser degree of intermittent plastic flow.

Mesoporous silica with cubic symmetry has attracted interest from researchers for some time. Here, we present the room temperature synthesis of mesoporous silica nanoparticles possessing cubic Pm3n symmetry with very high molar ratios (>50%) of 3-aminopropyl triethoxysilane. The synthesis is robust allowing, for example, co-condensation of organic dyes without loss of structure. By means of pore expander molecules, the pore size can be enlarged from 2.7 to 5 nm, while particle size decreases. Adding pore expander and co-condensing fluorescent dyes in the same synthesis reduces average particle size further down to 100 nm. After PEGylation, such fluorescent aminated mesoporous silica nanoparticles are spontaneously taken up by cells as demonstrated by fluorescence microscopy.

Synchrotron radiation (SR) sources have produced extraordinary advances in fields such as biology, chemistry, materials science, physics, and even in art and history. Storage-ring sources are continually being improved but have inherent limitations from the equilibrium emittances arising from many passes of the electron bunch around the ring. As we described in an earlier SRN article [1], an Energy Recovery Linac (ERL) source of X-rays can overcome many of these limitations to produce X-rays of high coherence and short pulse length, which can be focused down to a 1–10 nm round beam size.

Three research initiatives pursued by the Macromolecular Diffraction Facility at the Cornell High Energy Synchrotron Source (MacCHESS) are presented., The Macromolecular Diffraction Facility at the Cornell High Energy Synchrotron Source (MacCHESS) is a national research resource supported by the National Center for Research Resources of the US National Institutes of Health. MacCHESS is pursuing several research initiatives designed to benefit both CHESS users and the wider structural biology community. Three initiatives are presented in further detail: microcrystallography, which aims to improve the collection of diffraction data from crystals a few micrometers across, or small well diffracting regions of inhomogeneous crystals, so as to obtain high-resolution structures; pressure cryocooling, which can stabilize transient structures and reduce lattice damage during the cooling process; and BioSAXS (small-angle X-ray scattering on biological solutions), which can extract molecular shape and other structural information from macromolecules in solution.

We report the first use of a non-frustrated block terpolymer for the synthesis of highly ordered oxide nanocomposites containing multiple plies. The morphological behavior of 15 ISO-oxide nanocomposites was investigated spanning a large range of compositions along the ƒ(I)=ƒ(S) isopleth using aluminosilicate and niobia sols. Morphologies were determined by TEM and SAXS measurements. Four morphologies were identified, including core-shell hexagonal, core-shell double gyroid, three-domain lamellae, and core-shell inverse-hexagonal, in order of increasing O+oxide vol fraction. All of the resulting nanocomposites had three- or five-ply morphologies containing domains that were continuous in one, two, or three dimensions. The five-ply core-shell double gyroid phase was only found to be stable when the O+oxide domain was a minority. Removal of the polymer enabled simple and direct synthesis of mesoporous oxide materials while retaining the ordered network structure. We believe that advances in the synthesis of multi-ply nanocomposites will lead to advanced materials and devices containing multiple plies of functional materials.

We describe the development of a CMOS read-out chip for an X-ray pixel array detector that accumulates into analog storage elements with time resolution typical of synchrotron bunch spacings. Each pixel contains multiple analog storage elements to allow capture of full-frame images at submicrosecond separation. Additionally, each storage element is re-addressable, which allows accumulation of signal from temporally distinct acquisition windows. Test results show the ability to slew and settle the equivalent of 650 8-keV x-rays (1.21 Me-) in less than 100 ns. The detector RMS read-noise was measured to be 2350 e- and grows with the square-root of the number of accumulations with a coefficient of 415 e- (equivalent to 1.07 and 0.19 8-keV x-rays respectively). The saturation value for each storage element, in terms of 8 keV x-rays, was measured to exceed 1880 x-rays (6.42 Me-). The complete detector is anticipated to contain around 400 times 200 pixels with pixel size near 150 mum times 150 mum. Possible experimental applications include study of material failures, transient phase transformations, and high-speed X-radiography.

We report the first use of a block terpolymer for the synthesis of three-component nanocomposites. Here, the use of three chemically distinct polymer blocks enabled control over each of the three final components: partially graphitic carbon, crystalline transition metal oxide, and porosity. Tuning of the individual block lengths and block fractions resulted in control over the three components. Specifically, two PAN-b-PEO-b-PPO-b-PEO-b-PAN pentablock terpolymers were synthesized starting with a functionalized P123 or F127 macroinitiator. The PEO blocks were selectively swelled with titania sols while the PPO primarily served as a mesoporosity source and the PAN served as a carbon source with high yield. Two subsequent heat treatments were used to form partially graphitic carbon which acted as an in situ hard template preserving the mesostructure through the crystallization of the titania sols. TEM and SAXS analysis revealed wormlike microphase separation. Nitrogen physisorption analysis revealed that the ...