High-speed free-run ptychography at the Australian Synchrotron.

Over the last decade ptychography has progressed rapidly from a specialist ultramicroscopy technique into a mature method accessible to non-expert users. However, to improve scientific value ptychography data must reconstruct reliably, with high image quality and at no cost to other correlative methods. Presented here is the implementation of high-speed ptychography used at the Australian Synchrotron on the XFM beamline, which includes a free-run data collection mode where dead time is eliminated and the scan time is optimized. It is shown that free-run data collection is viable for fast and high-quality ptychography by demonstrating extremely high data rate acquisition covering areas up to 352 000 µm2 at up to 140 µm2 s-1, with 13× spatial resolution enhancement compared with the beam size. With these improvements, ptychography at velocities up to 250 µm s-1 is approaching speeds compatible with fast-scanning X-ray fluorescence microscopy. The combination of these methods provides morphological context for elemental and chemical information, enabling unique scientific outcomes.

A technique for preparing undecalcified osteochondral fresh frozen sections for elemental mapping and understanding disease etiology.

The anatomy of the osteochondral junction is complex because several tissue components exist as a unit, including uncalcified cartilage (with superficial, middle, and deep layers), calcified cartilage, and subchondral bone. Furthermore, it is difficult to study because this region is made up of a variety of cell types and extracellular matrix compositions. Using X-ray fluorescence microscopy, we present a protocol for simultaneous elemental detection on fresh frozen samples. We transferred the osteochondral sample using a tape-assisted system and successfully tested it in synchrotron X-ray fluorescence. This protocol elucidates the distinct distribution of elements at the human knee's osteochondral junction, making it a useful tool for analyzing the co-distribution of various elements in both healthy and diseased states.

Iterative energy self-calibration of Fe XANES spectra. Erratum.

A correction is made to the paper by Jones et al. (2020). [J. Synchrotron Rad. (2020), 27, 207-211].

Iterative energy self-calibration of Fe XANES spectra.

Determining the oxidation state of Fe through parameterization of X-ray absorption near-edge structure (XANES) spectral features is highly dependent on accurate and repeatable energy calibration between spectra. Small errors in energy calibration can lead to vastly different interpretations. While simultaneous measurement of a reference foil is often undertaken on X-ray spectroscopy beamlines, other beamlines measure XANES spectra without a reference foil and therefore lack a method for correcting energy drift. Here a method is proposed that combines two measures of Fe oxidation state taken from different parts of the spectrum to iteratively correct for an unknown energy offset between spectra, showing successful iterative self-calibration not only during individual beam time but also across different beamlines.

Assessing radiation dose limits for X-ray fluorescence microscopy analysis of plant specimens.

BACKGROUND AND AIMS: X-ray fluorescence microscopy (XFM) is a powerful technique to elucidate the distribution of elements within plants. However, accumulated radiation exposure during analysis can lead to structural damage and experimental artefacts including elemental redistribution. To date, acceptable dose limits have not been systematically established for hydrated plant specimens. METHODS: Here we systematically explore acceptable dose rate limits for investigating fresh sunflower (Helianthus annuus) leaf and root samples and investigate the time-dose damage in leaves attached to live plants. KEY RESULTS: We find that dose limits in fresh roots and leaves are comparatively low (4.1 kGy), based on localized disintegration of structures and element-specific redistribution. In contrast, frozen-hydrated samples did not incur any apparent damage even at doses as high as 587 kGy. Furthermore, we find that for living plants subjected to XFM measurement in vivo and grown for a further 9 d before being reimaged with XFM, the leaves display elemental redistribution at doses as low as 0.9 kGy and they continue to develop bleaching and necrosis in the days after exposure. CONCLUSIONS: The suggested radiation dose limits for studies using XFM to examine plants are important for the increasing number of plant scientists undertaking multidimensional measurements such as tomography and repeated imaging using XFM.

Monitoring compositional changes in Ni(OH)2 electrocatalysts employed in the oxygen evolution reaction.

Electrochemical water splitting to generate hydrogen has been identified as a possible solution to the storage of intermittent renewable energy. However there are still challenges remaining in the development of stable electrocatalysts for the oxygen evolution half-reaction. Here we investigate the effects that the oxygen evolution reaction (OER) has on an electrodeposited Ni(OH)2 catalyst operated under alkaline conditions. The electrocatalyst was characterised by established methods including cyclic voltammetry, electrochemical impedance spectroscopy, scanning electron microscopy, atomic force microscopy, X-ray photoelectron spectroscopy both before and after the OER to identify changes that may have occurred in the structure and/or composition of the catalyst. In addition, synchrotron X-ray absorption near edge structure mapping was used to generate spatially resolved maps of the species present within the Ni(OH)2 catalyst and how they change in a heterogeneous manner into a NiO species after the OER. When compared to the morphological data it suggests that changes in the morphology after the OER can be correlated to the formation of NiO within the newly formed clusters that were generated across the electrocatalyst.

Radiation Dose Limits for Bioanalytical X-ray Fluorescence Microscopy.

Analytical approaches that preserve the endogenous state of the examined system are essential for the in vivo study of bioinorganics. X-ray fluorescence microscopy of biological samples can map elements in vivo at subcellular resolutions in tissue samples and multicellular organisms. However, X-ray irradiation induces modifications that accumulate with dose. Consequently, the utility of X-ray fluorescence microscopy is intrinsically limited by the radiation damage it causes and the degree to which it alters the target features of interest. Identification of the dose threshold, below which the integrity of the specimen and its elemental distribution is preserved, is required to ensure valid interpretation of concentrations. Here we use the nematode, Caenorhabditis elegans, to explore these issues using three chemical-free specimen preparations: lyophilization, cryofixation, and live. We develop quantitative methods for investigating damage and present dose limits for each preparation pertaining to the micrometer-scale spatial distribution of specific analytes (potassium, calcium, manganese, iron, and zinc), and discuss dose-appropriate guidelines for X-ray fluorescence microscopy of microscale biological samples.

Copper-Catalyzed Synthesis and Applications of Yndiamides.

The first synthetic route to yndiamides, a novel class of double aza-substituted alkyne, has been established by the copper(I)-catalyzed cross-coupling of 1,1-dibromoenamides with nitrogen nucleophiles. The utility of these compounds is demonstrated in a range of transition-metal-catalyzed and acid-catalyzed transformations to afford a wide variety of 1,2-diamide functionalized products.

Simultaneous X-ray fluorescence and scanning X-ray diffraction microscopy at the Australian Synchrotron XFM beamline.

Owing to its extreme sensitivity, quantitative mapping of elemental distributions via X-ray fluorescence microscopy (XFM) has become a key microanalytical technique. The recent realisation of scanning X-ray diffraction microscopy (SXDM) meanwhile provides an avenue for quantitative super-resolved ultra-structural visualization. The similarity of their experimental geometries indicates excellent prospects for simultaneous acquisition. Here, in both step- and fly-scanning modes, robust, simultaneous XFM-SXDM is demonstrated.

Exploiting orientation-selective DEER: determining molecular structure in systems containing Cu(ii) centres.

Orientation-selective DEER (Double Electron-Electron Resonance) measurements were conducted on a series of rigid and flexible molecules containing Cu(ii) ions. A system with two rigidly held Cu(ii) ions was afforded by the protein homo-dimer of copper amine oxidase from Arthrobacter globiformis. This system provided experimental DEER data between two Cu(ii) ions with a well-defined distance and relative orientation to assess the accuracy of the methodology. Evaluation of orientation-selective DEER (os DEER) on systems with limited flexibility was probed using a series of porphyrin-based Cu(ii)-nitroxide and Cu(ii)-Cu(ii) model systems of well-defined lengths synthesized for this project. Density functional theory was employed to generate molecular models of the conformers for each porphyrin-based Cu(ii) dimer studied. Excellent agreement was found between DEER traces simulated using these computed conformers and the experimental data. The performance of different parameterised structural models in simulating the experimental DEER data was also investigated. The results of this analysis demonstrate the degree to which the DEER data define the relative orientation of the two Cu(ii) ions and highlight the need to choose a parameterised model that captures the essential features of the flexibility (rotational freedom) of the system being studied.

Elemental mapping of the entire intact Drosophila gastrointestinal tract.

The main role of the animal gastrointestinal (GI) tract is the selective absorption of dietary nutrients from ingested food sources. One class of vital micronutrients are the essential biometals such as copper, zinc and iron, which participate in a plethora of biological process, acting as enzymatic or structural co-factors for numerous proteins and also as important cellular signalling molecules. To help elucidate the mechanisms by which biometals are absorbed from the diet, we mapped elemental distribution in entire, intact Drosophila larval GI tracts using synchrotron X-ray fluorescence microscopy. Our results revealed distinct regions of the GI tract enriched for specific metals. Copper was found to be concentrated in the copper cell region but also in the region directly anterior to the copper cells and unexpectedly, in the middle midgut/iron cell region as well. Iron was observed exclusively in the iron cell region, confirming previous work with iron-specific histological stains. Zinc was observed throughout the GI tract with an increased accumulation in the posterior midgut region, while manganese was seen to co-localize with calcium specifically in clusters in the distal Malpighian tubules. This work simultaneously reveals distribution of a number of biologically important elements in entire, intact GI tracts. These distributions revealed not only a previously undescribed Ca/Mn co-localization, but also the unexpected presence of additional Cu accumulations in the iron cell region.

Kinetically Stable Lanthanide Complexes Displaying Exceptionally High Quantum Yields upon Long-Wavelength Excitation: Synthesis, Photophysical Properties, and Solution Speciation.

We demonstrate how highly emissive, kinetically stable complexes can be prepared using the macrocyclic scaffold of DO3A bearing coordinating aryl ketones as highly effective sensitizing chromophores. In the europium complexes, high quantum yields (up to 18% in water) can be combined with long-wavelength excitation (370 nm). The behavior in solution upon variation of pH, studied by means of UV-vis absorption, emission, and NMR spectroscopies, reveals that the nature of the chromophore can give rise to pH-dependent behavior as a consequence of deprotonation adjacent to the carbonyl group. Knowledge of the molecular speciation in solution is therefore critical when assessing the luminescence properties of such complexes.

The role of plants in ironstone evolution: iron and aluminium cycling in the rhizosphere.

The Carajás plateaus in Brazil host endemic epilithic vegetation ("campo rupestre") on top of ironstone duricrusts, known as canga. This capping rock is primarily composed of iron(III) oxide minerals and forms a physically resistant horizon. Field observations reveal an intimate interaction between canga's surface and two native sedges (Rhynchospora barbata and Bulbostylis cangae). These observations suggest that certain plants contribute to the biogeochemical cycling of iron. Iron dissolution features at the root-rock interface were characterised using synchrotron-based techniques, Raman spectroscopy and scanning electron microscopy. These microscale characterisations indicate that iron is preferentially leached in the rhizosphere, enriching the comparatively insoluble aluminium around root channels. Oxalic acid and other exudates were detected in active root channels, signifying ligand-controlled iron oxide dissolution, likely driven by the plants' requirements for goethite-associated nutrients such as phosphorus. The excess iron not uptaken by the plant can reprecipitate in and around roots, line root channels and cement detrital fragments in the soil crust at the base of the plants. The reprecipitation of iron is significant as it provides a continuously forming cement, which makes canga horizons a 'self-healing' cover and contributes to them being the world's most stable continuously exposed land surfaces. Aluminium hydroxide precipitates ("gibbsite cutans") were also detected, coating some of the root cavities, often in alternating layers with goethite. This alternating pattern may correspond with oscillating oxygen concentrations in the rhizosphere. Microbial lineages known to contain iron-reducing bacteria were identified in the sedge rhizospheric microbiome and likely contribute to the reductive dissolution of iron(III) oxides within canga. Drying or percolation of oxygenated water to these anaerobic niches have led to iron mineralisation of biofilms, detected in many root channels. This study sheds light on plants' direct and indirect involvement in canga evolution, with possible implications for revegetation and surface restoration of iron mine sites.

Placental Element Content Assessed via Synchrotron-Based X-ray Fluorescence Microscopy Identifies Low Molybdenum Concentrations in Foetal Growth Restriction, Postdate Delivery and Stillbirth.

Placental health and foetal development are dependent upon element homeostasis. Analytical techniques such as mass spectroscopy can provide quantitative data on element concentrations in placental tissue but do not show spatial distribution or co-localisation of elements that may affect placental function. The present study used synchrotron-based X-ray fluorescence microscopy to elucidate element content and distribution in healthy and pathological placental tissue. The X-ray fluorescence microscopy (XFM) beamline at the Australian Synchrotron was used to image trace metal content of 19 placental sections from healthy term (n = 5, 37-39 weeks), foetal growth-restricted (n = 3, <32 weeks, birth weight <3rd centile), postdate (n = 7, >41 completed weeks), and stillbirth-complicated pregnancies (n = 4, 37-40 weeks). Samples were cryo-sectioned and freeze-dried. The concentration and distribution of fourteen elements were detected in all samples: arsenic, bromine, calcium, chlorine, copper, iron, molybdenum, phosphorous, potassium, rubidium, selenium, strontium, sulphur, and zinc. The elements zinc, calcium, phosphorous, and strontium were significantly increased in stillbirth placental tissue in comparison to healthy-term controls. Strontium, zinc, and calcium were found to co-localise in stillbirth tissue samples, and calcium and strontium concentrations were correlated in all placental groups. Molybdenum was significantly decreased in stillbirth, foetal growth-restricted, and postdate placental tissue in comparison to healthy-term samples (p < 0.0001). Synchrotron-based XFM reveals elemental distribution within biological samples such as the placenta, allowing for the co-localisation of metal deposits that may have a pathological role. Our pilot study further indicates low concentrations of placental molybdenum in pregnancies complicated by foetal growth restriction, postdate delivery, and stillbirth.

Operando Investigation of Silicon Anodes During Electrochemical Cycling in Li-ion Batteries.

Silicon (Si) is recognized as a promising anode material for next-generation anodes due to its high capacity. However, large volume expansion and active particle pulverization during cycling rapidly deteriorate the battery performance. The relationship between Si anode particle size and particle pulverization, and the structure evolution of Si particles during cycling is not well understood. In this study, a quantitative, time-resolved "operando" small angle X-ray scattering (SAXS) investigation into the morphological change of unwrapped and reduced graphene oxide (rGO) wrapped Si nanoparticles (Si@rGO) is conducted with respect to the operating voltage. The results provide a clear picture of Si particle size change and the role of nonrigid rGO in mitigating Si volume expansion and pulverization. Further, this study demonstrates the advantage of "operando" SAXS in electrochemical environments as compared to other approaches.

Using remote substituents to control solution structure and anion binding in lanthanide complexes.

A study of the anion-binding properties of three structurally related lanthanide complexes, which all contain chemically identical anion-binding motifs, has revealed dramatic differences in their anion affinity. These arise as a consequence of changes in the substitution pattern on the periphery of the molecule, at a substantial distance from the binding pocket. Herein, we explore these remote substituent effects and explain the observed behaviour through discussion of the way in which remote substituents can influence and control the global structure of a molecule through their demands upon conformational space. Peripheral modifications to a binuclear lanthanide motif derived from α,α'-bis(DO3 Ayl)-m-xylene are shown to result in dramatic changes to the binding constant for isophthalate. In this system, the parent compound displays considerable conformational flexibility, yet can be assumed to bind to isophthalate through a well-defined conformer. Addition of steric bulk remote from the binding site restricts conformational mobility, giving rise to an increase in binding constant on entropic grounds as long as the ideal binding conformation is not excluded from the available range of conformers.

Synthesis of rac-Lindenene via a thermally induced cyclopropanation reaction.

The first synthesis of the sesquiterpene Lindenene is described. A novel non-catalysed intramolecular cyclopropanation reaction between a diazoketone and an unactivated alkene was utilised to construct the relatively labile ketone precursor with complete stereocontrol. This ketone was transformed in three steps into Lindenene.

Fluorinated quinazolinones as potential radiotracers for imaging kinesin spindle protein expression.

Anti-mitotic anti-cancer drugs offer a potential platform for developing new radiotracers for imaging proliferation markers associated with the mitosis-phase of the cell-cycle. One interesting target is kinesin spindle protein (KSP)-an ATP-dependent motor protein that plays a vital role in bipolar spindle formation. In this work we synthesised a range of new fluorinated-quinazolinone compounds based on the structure of the clinical candidate KSP inhibitor, ispinesib, and investigated their properties in vitro as potential anti-mitotic agents targeting KSP expression. Anti-proliferation (MTT and BrdU) assays combined with additional studies including fluorescence-assisted cell sorting (FACS) analysis of cell-cycle arrest confirmed the mechanism and potency of these biphenyl compounds in a range of human cancer cell lines. Additional studies using confocal fluorescence microscopy showed that these compounds induce M-phase arrest via monoaster spindle formation. Structural studies revealed that compound 20-(R) is the most potent fluorinated-quinazolinone inhibitor of KSP and represents a suitable lead candidate for further studies on designing (18)F-radiolabelled agents for positron-emission tomography (PET).

Balance-based exercise programs on balance in older adults with mild to moderate dementia: A critically appraised topic.

OBJECTIVE: It is well established that exercise programs including balanced-based exercises are practical and beneficial for cognitively healthy older adults. However, there is limited evidence to determine if these types of training regimens are effective for individuals with dementia. We conducted a critical appraisal to determine if the addition of balance-based exercise programs, compared with usual care at residential homes, improved balance in adults diagnosed with mild to moderate dementia. MATERIALS AND METHODS: Four databases were searched for randomized control trials implementing balanced-based exercise programs with individuals diagnosed with mild to moderate dementia. The articles were evaluated based on the Centre for Evidence Based Medicine level of evidence criteria and appraised using the PEDro scale. RESULTS: A comprehensive literature search revealed four studies relevant to the clinical question that fit the inclusion and exclusion criteria. All four studies indicated an improvement in balance following a weekly physical therapy program implementing balance-based exercises. CONCLUSIONS: There is sufficient evidence to suggest that the use of physical therapy programs that include balance-based exercises, performed 1-2 times a week over the course of 12-25 weeks, improves balance in elderly adults with mild to moderate dementia.

Microbially influenced tungsten mobilization and formation of secondary minerals in wolframite tailings.

Wolframite [(Fe,Mn)WO4] tailings represent a hazardous waste that can pose a threat to the environment, humans, animals and plants. The present study aims to conduct a high-resolution depth profile characterization of wolframite tailings from Wolfram Camp, North Queensland, Australia, to understand the biogeochemical influences on W mobilization. Several indigenous Fe- and S-oxidizing bacteria (e.g., Streptococcus pneumoniae and Thiomonas delicata) in wolframite tailings were found highly associated with W, As, and rare earth elements. Biooxidation of metal sulfides, i.e., pyrite, molybdenite and bismuthinite, produced sulfuric acid, which accelerated the weathering of wolframite, mobilizing tungstate (WO42-). Using synchrotron-based X-ray fluorescence microscopy (XFM) and W L-edge X-ray absorption near-edge spectroscopy (µ-XANES) analysis, wolframite was initially transformed into Na- and Bi- tungstate as well as tungstic acid (partial weathering) followed by the formation of Ga- and Zn- tungstate after extensive weathering, i.e., the wolframite had disappeared. While W (VI) was the major W species in wolframite tailings, minor W(0) and W(II), and trace W(IV) were also detected. The major contaminant in the Wolfram Camp tailings was As. Though wolframite tailings are hazardous waste, the toxicity of W was unclear. Tungsten waste still has industrial value; apart from using them as substitution material for cement and glass production, there is interest in reprocessing W waste for valuable metal recovery. If the environmental benefits are taken into consideration, i.e., preventing the release of toxic metals into surrounding waterways, reprocessing may be economic.