Utilizing Synchrotron-Based X-ray Micro-Computed Tomography to Visualize the Microscopic Structure of Starch Hydrogels In Situ.

This study aimed to visualize the microstructures of starch hydrogels using synchrotron-based X-ray micro-computed tomography (µCT). Waxy maize starch (WMS, 3.3% amylose, db), pea starch (PS, 40.3% amylose), and high-amylose maize starch (HMS, 63.6% amylose) were cooked at 95 and 140 °C to prepare starch hydrogels. WMS and HMS failed to form a gel after 95 °C cooking and storage, while PS developed a firm gel. At 140 °C cooking, HMS of a high amylose nature was fully gelatinized and generated a rigid gel with the highest strength. Both scanning electron microscopy (SEM) and µCT revealed the unique structural features of various starch hydrogels/pastes prepared at different temperatures, which were greatly affected by the degree of swelling and dispersity of the starches. As a nondestructive method, µCT showed certain advantages over SEM, including minimal shrinkage of the hydrogels, relatively simple sample preparation, and allowing for three-dimensional reconstruction of the hydrogel microstructure. This study indicated that synchrotron-based µCT could be a useful technique in visualizing biopolymer-based hydrogels.

Nutri-cereal tissue-specific transcriptome atlas during development: Functional integration of gene expression to identify mineral uptake pathways in little millet (Panicum sumatrense).

Little millet (Panicum sumatrense Roth ex Roem. & Schult.) is an essential minor millet of southeast Asia and Africa's temperate and subtropical regions. The plant is stress-tolerant, has a short life cycle, and has a mineral-rich nutritional profile associated with unique health benefits. We report the developmental gene expression atlas of little millet (genotype JK-8) from ten tissues representing different stages of its life cycle, starting from seed germination and vegetative growth to panicle maturation. The developmental transcriptome atlas led to the identification of 342 827 transcripts. The BUSCO analysis and comparison with the transcriptomes of related species confirm that this study presents high-quality, in-depth coverage of the little millet transcriptome. In addition, the eFP browser generated here has a user-friendly interface, allowing interactive visualizations of tissue-specific gene expression. Using these data, we identified transcripts, the orthologs of which in Arabidopsis and rice are involved in nutrient acquisition, transport, and response pathways. The comparative analysis of the expression levels of these transcripts holds great potential for enhancing the mineral content in crops, particularly zinc and iron, to address the issue of "hidden hunger" and to attain nutritional security, making it a valuable asset for translational research.

Using BpyAla to generate copper artificial metalloenzymes: a catalytic and structural study.

Artificial metalloenzymes (ArMs) have emerged as a promising avenue in the field of biocatalysis, offering new reactivity. However, their design remains challenging due to the limited understanding of their protein dynamics and how the introduced cofactors alter the protein scaffold structure. Here we present the structures and catalytic activity of novel copper ArMs capable of (R)- or (S)-stereoselective control, utilizing a steroid carrier protein (SCP) scaffold. To incorporate 2,2'-bipyridine (Bpy) into SCP, two distinct strategies were employed: either Bpy was introduced as an unnatural amino acid (2,2'-bipyridin-5-yl)alanine (BpyAla) using amber stop codon expression or via bioconjugation of bromomethyl-Bpy to cysteine residues. The resulting ArMs proved to be effective at catalysing an enantioselective Friedel-Crafts reaction with SCP_Q111BpyAla achieving the best selectivity with an enantioselectivity of 72% ee (S). Interestingly, despite using the same protein scaffold, different attachment strategies for Bpy at the same residue (Q111) led to a switch in the enantiopreference of the ArM. X-ray crystal structures of SCP_Q111CBpy and SCP_Q111BpyAla ArMs with bound Cu(ii) ions unveiled crucial differences in the orientation of the catalytic centre. Combining structural information, alanine scanning studies, and computational analysis shed light on the distinct active sites of the ArMs, clarifying that these active sites stabilise the nucleophilic substrate on different sides of the electrophile leading to the observed switch in enantioselectivity. This work underscores the importance of integrating structural studies with catalytic screening to unravel the intricacies of ArM behaviour and facilitate their development for targeted applications in biocatalysis.

Drought-induced fiber water release and xylem embolism susceptibility of intact balsam poplar saplings.

Balsam poplar (Populus balsamifera L.) is a widespread tree species in North America with significant ecological and economic value. However, little is known about the susceptibility of saplings to drought-induced embolism and its link to water release from surrounding xylem fibers. Questions remain regarding localized mechanisms that contribute to the survival of saplings in vivo of this species under drought. Using X-ray micro-computed tomography on intact saplings of genotypes Gillam-5 and Carnduff-9, we found that functional vessels are embedded in a matrix of water-filled fibers under well-watered conditions in both genotypes. However, water-depleted fibers started to appear under moderate drought stress while vessels remained water-filled in both genotypes. Drought-induced xylem embolism susceptibility was comparable between genotypes, and a greater frequency of smaller diameter vessels in GIL-5 did not increase embolism resistance in this genotype. Despite having smaller vessels and a total vessel number that was comparable to CAR-9, stomatal conductance was generally higher in GIL-5 compared to CAR-9. In conclusion, our in vivo data on intact saplings indicate that differences in embolism susceptibility are negligible between GIL-5 and CAR-9, and that fiber water release should be considered as a mechanism that contributes to the maintenance of vessel functional status in saplings of balsam poplar experiencing their first drought event.

Synchrotron micro-computed tomography unveils the three-dimensional structure and origin of staminodes in the Plains Prickly Pear Cactus Opuntia polyacantha Haw. (Cactaceae).

Floral appendages display an array of shapes and sizes. Among these organs, staminodes are morphologically diverse structures that have lost the ability to produce pollen, but in some instances, they produce fertile pollen grains. In the family Cactaceae staminodes are uncommon and range from simple linear to flat to spatulate structures, but studies describing their structural attributes are scanty. This study highlights the advantages of synchrotron radiation for sample preparation and as a research tool for plant biology. It describes the internal morphology of floral parts, particularly stamen, tepal, and staminode in the Plains Prickly Pear Cactus, Opuntia polyacantha, using synchrotron radiation micro-computed tomography (SR-µCT). It also shows the different anatomical features in reconstructed three-dimensional imaging of reproductive parts and discuss the advantages of the segmentation method to detect and characterize the configuration and intricate patterns of vascular networks and associated structures of tepal and androecial parts applying SR-µCT. This powerful technology led to substantial improvements in terms of resolution allowing a more comprehensive understanding of the anatomical organization underlying the vasculature of floral parts and inception of staminodes in O. polyacantha. Tepal and androecial parts have uniseriate epidermis enclosing loose mesophyll with mucilage secretory ducts, lumen, and scattered vascular bundles. Cryptic underlying structural attributes provide evidence of a vascularized pseudo-anther conjoint with tepals. The undefined contours of staminodial appendages (pseudo-anther) amalgamated to the tepals' blurred boundaries suggest that staminodes originate from tepals, a developmental pattern supporting the fading border model of floral organ identity for angiosperms.

Tempering of cocoa butter and chocolate using minor lipidic components.

Chocolate manufacture includes a complex tempering procedure to direct the crystallization of cocoa butter towards the formation of fat crystal networks with specific polymorphism, nano- and microstructure, melting behavior, surface gloss and mechanical properties. Here we investigate the effects of adding various minor non-triglyceride lipidic components to refined cocoa butter and chocolate on their physical properties. We discover that addition of saturated phosphatidylcholine and phosphatidylethanolamine to neutralized and bleached cocoa butter or molten and recrystallized commercial chocolate at 0.1% (w/w) levels, followed by rapid cooling to 20 °C in the absence of shear, accelerates crystallization, stabilizes the desirable Form V polymorph and induces the formation of chocolate with an optimal microstructure, surface gloss and mechanical strength. Final chocolate structure and properties are comparable to those of a commercial tempered chocolate. Minor lipidic component addition represents an effective way to engineer chocolate material properties at different length scales, thus simplifying the entire tempering process.

The experience of using prompting technology from the perspective of people with Dementia and their primary carers.

OBJECTIVES: People who are living with dementia typically experience difficulties in completing multi-step, everyday tasks. However, digital technology such as touchscreen tablets provide a means of delivering concise personalised prompts that combine audio, text and pictures. This study was one component of a broader, mixed methods study that tested how an application (app) -based prompter running on a touchscreen tablet computer could support everyday activities in individuals with mild to moderate dementia. In this study we set out to understand the experiences of people living with dementia and their primary carer in using the prompter over a four-week period. METHOD: We collected qualitative data using semi-structured interviews from 26 dyads, composed of a person living with dementia and their carer. Dyads were interviewed at the start and end of this period. Transcripts were then analysed using thematic analysis. RESULTS: The study identified three overarching themes related to: participants' attitudes towards the technology; their judgements about how useful the prompter would be; and the emotional impact of using it. CONCLUSION: Consistent with the Technology Acceptance Model, carers and participants were influenced by their approaches to technology and determined the usefulness of the prompter according to whether it worked for them and fitted into their routines. In addition, participants' decisions about using the prompter were also determined by the extent to which doing so would impact on their self-identity.

Effect of an extreme flood event on solute transport and resilience of a mine water treatment system in a mineralised catchment.

Extreme rainfall events are predicted to become more frequent with climate change and can have a major bearing on instream solute and pollutant transport in mineralised catchments. The Coledale Beck catchment in north-west England was subject to an extreme rainfall event in December 2015 that equated to a 1 in 200-year event. The catchment contains the UK's first passive metal mine water treatment system, and as such had been subject to intense monitoring of solute dynamics before and after commissioning. Due to this monitoring record, the site provides a unique opportunity to assess the effects of a major storm event on (1) catchment-scale solute transport, and (2) the resilience of the new and novel passive treatment system to extreme events. Monitoring suggests a modest decline in treatment efficiency over time that is not synchronous with the storm event and explained instead by changes in system hydraulic efficiency. There was no apparent flushing of the mine system during the event that could potentially have compromised treatment system performance. Analysis of metal transport in the catchment downstream of the mine suggests relatively subtle changes in instream chemistry with modest but statistically-significant reductions in zinc in the lower catchment irrespective of flow condition after the extreme event, but most parameters of interest show no significant change. Increased export of colloidal iron and aluminium is associated with major landslips in the mid-catchment after the storm and provide fresh sorption sites to attenuate dissolved zinc more rapidly in these locations, corroborated by laboratory experiments utilising site materials to investigate the attenuation/release of metals from stream and terrestrial sediments. The data are important as they show both the resilience of passive mine water treatment systems to extreme events and the importance of catchment-scale monitoring to ensure continued effectiveness of treatment initiatives after major perturbation.

Fracture risk is under-recognised and under-treated in memory clinic attendees.

UK national guidelines recommend that older people at risk of falling should have their fracture risk assessed and acted upon. People with cognitive impairment are more likely to sustain a fracture than their cognitively intact peers. We assessed the fracture risk of 79 memory clinic attendees and compared their actual management with guidelines. Despite reporting 57 falls in the last year, only 36% of those who would be recommended antiresportive treatment were prescribed it and a dual-energy X-ray absorptiometry (DEXA)scan was performed in only 13% where it would be recommended. These findings highlight an important deficit in fracture risk assessment which should inform future interventions.

Geochemical recovery of the Torna-Marcal river system after the Ajka red mud spill, Hungary.

The failure of the Ajka red mud depository in October 2010 led to the largest single release of red mud into the surface water environment. This study provides a comparative assessment of stream sediment quality in the Torna-Marcal-Rába catchment between post-disaster surveys (2010) and follow up surveys at an identical suite of 21 locations in 2013. The signature of red mud apparent in initial surveys with high Al, As, Cr, Na, V was only apparent at a small number of sample stations in recent surveys. These constitute <1 km of stream, compared to the >20 km reach of affected sediments in the immediate aftermath of the spill. Concentrations of red mud-derived contaminants are predominately associated with fine fractions of the red mud (<8 µm). This enhances transport out of the system of red mud-derived contaminants and, along with extensive remedial efforts, has substantially limited the within-channel inventory of potentially ecotoxic metals and metalloids.

Food security in a perfect storm: using the ecosystem services framework to increase understanding.

Achieving food security in a 'perfect storm' scenario is a grand challenge for society. Climate change and an expanding global population act in concert to make global food security even more complex and demanding. As achieving food security and the millennium development goal (MDG) to eradicate hunger influences the attainment of other MDGs, it is imperative that we offer solutions which are complementary and do not oppose one another. Sustainable intensification of agriculture has been proposed as a way to address hunger while also minimizing further environmental impact. However, the desire to raise productivity and yields has historically led to a degraded environment, reduced biodiversity and a reduction in ecosystem services (ES), with the greatest impacts affecting the poor. This paper proposes that the ES framework coupled with a policy response framework, for example Driver-Pressure-State-Impact-Response (DPSIR), can allow food security to be delivered alongside healthy ecosystems, which provide many other valuable services to humankind. Too often, agro-ecosystems have been considered as separate from other natural ecosystems and insufficient attention has been paid to the way in which services can flow to and from the agro-ecosystem to surrounding ecosystems. Highlighting recent research in a large multi-disciplinary project (ASSETS), we illustrate the ES approach to food security using a case study from the Zomba district of Malawi.

Trajectory Synthesis for Fisher Information Maximization.

Estimation of model parameters in a dynamic system can be significantly improved with the choice of experimental trajectory. For general nonlinear dynamic systems, finding globally "best" trajectories is typically not feasible; however, given an initial estimate of the model parameters and an initial trajectory, we present a continuous-time optimization method that produces a locally optimal trajectory for parameter estimation in the presence of measurement noise. The optimization algorithm is formulated to find system trajectories that improve a norm on the Fisher information matrix (FIM). A double-pendulum cart apparatus is used to numerically and experimentally validate this technique. In simulation, the optimized trajectory increases the minimum eigenvalue of the FIM by three orders of magnitude, compared with the initial trajectory. Experimental results show that this optimized trajectory translates to an order-of-magnitude improvement in the parameter estimate error in practice.

Thrombin generation and fibrin formation under flow on biomimetic tissue factor-rich surfaces.

BACKGROUND: Blood flow regulates coagulation and fibrin assembly by controlling the rate of transport of zymogens, enzymes and plasma proteins to and from the site of an injury. OBJECTIVE: The objective of this work was to define the hemodynamic conditions under which fibrin can form under flow on tissue factor (TF)-rich substrates. METHODS: TF-coated silica beads (~ 800 nm) were patterned into 18-85-µm spots. Normal pooled plasma and factors VIII, IX and XI deficient plasmas were perfused over the beads coated with 0.08, 0.8 and 8 molecules-TF µm(-2) at shear rates of 50-1000 s(-1) . Fibrin deposition and thrombin generation were measured by fluorescence microscopy in a hydrodynamic focusing microfluidic device. RESULTS AND CONCLUSIONS: Fibrin deposition was supported on patterned bead spots, but not planar TF substrates at the same surface TF concentration. There was a threshold spot size and a shear rate dependent TF concentration that was necessary to support fibrin polymerization. FVIII and FIX had minor effects on fibrin dynamics at 8 molecules-TF µm(-2) , but were essential at 0.8 molecules-TF µm(-2) . The absence of FXI influenced thrombin generation and fibrin deposition at both 0.8 and 8 molecules-TF µm(-2) . These results show that fibrin deposition requires perturbations in the flow field that protect reactions from dilution by flow under venous and arterial conditions. FVIII and FIX have a modest effect on fibrin deposition at high TF concentrations, but are necessary for fibrin deposition at low TF concentrations. FXI amplifies thrombin generation under flow at both low and high TF concentrations.

Application of the distributed point source method to rough surface scattering and ultrasonic wall thickness measurement.

The distributed point source method is commonly used to predict the complex acoustic field emitted by ultrasonic transducers. In this paper, it is presented as an alternative to conventional approaches often used when solving rough surface scattering problems. Surface shadowing and multiple scattering effects are inherently included in the mesh-free semi-analytical simulation method through matrix manipulation making it very efficient and simple to implement. Results are presented which illustrate the improvement in accuracy gained over the Kirchhoff approximation and the decrease in computational load over the finite element method, culminating in greater than an order of magnitude decrease in required simulation time. The method is applied to the practical problem of online wall thickness monitoring within corrosive environments, illustrating the variability in reflected pulse shape that could be expected from rough surfaces with similar statistics. Three commonly implemented time-of-flight algorithms are used to analyze a large number of simulated signals from which it is concluded that those based on first arrival time are more stable under increasing roughness conditions than those which are based on reflected pulse shape.

Contaminant mobility and carbon sequestration downstream of the Ajka (Hungary) red mud spill: The effects of gypsum dosing.

A number of emergency pollution management measures were enacted after the accidental release of caustic bauxite processing residue that occurred in Ajka, western Hungary in October, 2010. These centred on acid and gypsum dosing to reduce pH and minimise mobility of oxyanion contaminants mobile at high pH. This study assessed the effectiveness of gypsum dosing on contaminant mobility and carbon sequestration through assessment of red mud and gypsum-affected fluvial sediments via elemental analysis and stable isotope analysis. There was a modest uptake of contaminants (notably As, Cr, and Mn) on secondary carbonate-dominated deposits in reaches subjected to gypsum dosing. C and O stable isotope ratios of carbonate precipitates formed as a result of gypsum dosing were used to quantify the importance of the neutralisation process in sequestering atmospheric carbon dioxide. This process was particularly pronounced at sites most affected by gypsum addition, where up to 36% of carbonate-C appears to be derived from atmospheric in-gassing of CO(2). The site is discussed as a large scale analogue for potential remedial approaches and carbon sequestration technologies that could be applied to red mud slurries and other hyperalkaline wastes. The results of this work have substantial implications for the aluminium production industry in which 3-4% of the direct CO(2) emissions may be offset by carbonate precipitation. Furthermore, carbonation by gypsum addition may be important for contaminant remediation, also providing a physical stabilisation strategy for the numerous historic stockpiles of red mud.

Diurnal fluctuation of zinc concentration in metal polluted rivers and its potential impact on water quality and flux estimates.

Diurnal fluctuations of metals have been observed in the South Tyne river catchment, UK, in both upland tributaries and major river reaches. Zinc exhibits the most pronounced cyclicity, with concentrations increasing during the night to a maximum near 05:00 before decreasing during the day. This trend is the inverse of pH and temperature observations, which are thought to be the predominant drivers behind the cyclicity. Photosynthetic reactions of biomass and algae alter the pH within the river systems, therefore promoting hydrous metal oxide precipitation during the daylight which consequently allows divalent cations including zinc to sorb onto them. This mechanism may be supported by direct uptake of zinc by algae and other biogeochemical reactions which combine to create large differences in zinc concentrations; during base flow zinc concentrations increased by 326% from the minima over 48 hours. Maximum concentrations are not being captured during routine water quality analysis resulting in inaccurate and misleading EQS results and total flux estimations, for example the annual total zinc flux in a small tributary increases from 17 to 76 tonnes/year when routine grab sample data are supplemented with 24-hour sampling results.

Treatment of zinc-rich acid mine water in low residence time bioreactors incorporating waste shells and methanol dosing.

Bioreactors utilising bacterially mediated sulphate reduction (BSR) have been widely tested for treating metal-rich waters, but sustained treatment of mobile metals (e.g. Zn) can be difficult to achieve in short residence time systems. Data are presented providing an assessment of alkalinity generating media (shells or limestone) and modes of metal removal in bioreactors receiving a synthetic acidic metal mine discharge (pH 2.7, Zn 15 mg/L, SO(4)(2-) 200mg/L, net acidity 103 mg/L as CaCO(3)) subject to methanol dosing. In addition to alkalinity generating media (50%, v.v.), the columns comprised an organic matrix of softwood chippings (30%), manure (10%) and anaerobic digested sludge (10%). The column tests showed sustained alkalinity generation, which was significantly better in shell treatments. The first column in each treatment was effective throughout the 422 days in removing >99% of the dissolved Pb and Cu, and effective for four months in removing 99% of the dissolved Zn (residence time: 12-14 h). Methanol was added to the feedstock after Zn breakthrough and prompted almost complete removal of dissolved Zn alongside improved alkalinity generation and sulphate attenuation. While there was geochemical evidence for BSR, sequential extraction of substrates suggests that the bulk (67-80%) of removed Zn was associated with Fe-Mn oxide fractions.

Seasonal and spatial variation of diffuse (non-point) source zinc pollution in a historically metal mined river catchment, UK.

Quantifying diffuse sources of pollution is becoming increasingly important when characterising river catchments in entirety - a prerequisite for environmental management. This study examines both low and high flow events, as well as spatial variability, in order to assess point and diffuse components of zinc pollution within the River West Allen catchment, which lies within the northern England lead-zinc Orefield. Zinc levels in the river are elevated under all flow regimes, and are of environmental concern. Diffuse components are of little importance at low flow, with point source mine water discharges dominating instream zinc concentration and load. During higher river flows 90% of the instream zinc load is attributed to diffuse sources, where inputs from resuspension of metal-rich sediments, and groundwater influx are likely to be more dominant. Remediating point mine water discharges should significantly improve water quality at lower flows, but contribution from diffuse sources will continue to elevate zinc flux at higher flows.

Hydraulic residence time and iron removal in a wetland receiving ferruginous mine water over a 4 year period from commissioning.

Analysis of residence time distribution (RTD) has been conducted for the UK Coal Authority's mine water treatment wetland at Lambley, Northumberland, to determine the hydraulic performance of the wetland over a period of approximately 4 years since site commissioning. The wetland RTD was evaluated in accordance with moment analysis and modelled based on a tanks-in-series (TIS) model to yield the hydraulic characteristics of system performance. Greater hydraulic performance was seen during the second site monitoring after 21 months of site operation i.e. longer hydraulic residence time to reflect overall system hydraulic efficiency, compared to wetland performance during its early operation. Further monitoring of residence time during the third year of wetland operation indicated a slight reduction in hydraulic residence time, thus a lower system hydraulic efficiency. In contrast, performance during the fourth year of wetland operation exhibited an improved overall system hydraulic efficiency, suggesting the influence of reed growth over the lifetime of such systems on hydraulic performance. Interestingly, the same pattern was found for iron (which is the primary pollutant of concern in ferruginous mine waters) removal efficiency of the wetland system from the second to fourth year of wetland operation. This may therefore, reflect the maturity of reeds for maintaining efficient flow distribution across the wetland to retain a longer residence time and significant fractions of water involved to enhance the extent of treatment received for iron attenuation. Further monitoring will be conducted to establish whether such performance is maintained, or whether efficiency decreases over time due to accumulation of dead plant material within the wetland cells.

Inventory of aquatic contaminant flux arising from historical metal mining in England and Wales.

The impact of discharges from abandoned metal and ironstone mines has been a much studied form of aquatic pollution in recent decades. Few attempts however, have been made to accurately determine the overall contaminant mass flux arising from abandoned mine sites at scales above catchment level. Such assessments are critical to determine the significance of former mining to national, regional and ultimately global trace metal flux. This paper presents the most comprehensive national survey to date across England and Wales of the total pollution burden discharged at source from abandoned non-coal mine sites. 338 discharges have been identified (from 4923 known abandoned metal mines) and while concurrent flow and contaminant concentration records are only available for around 30% of these, significant quantities of metals (and As) have been quantified to be discharged. A minimum of 193 tonnes of Zn, 18.5 tonnes of Pb, 0.64 tonnes Cd, 19.1 tonnes of Cu, 551 tonnes Fe, 72 tonnes Mn and 5.1 tonnes As are released in water discharges from abandoned non-coal mines to the surface water environment of England and Wales each year. Precautionary extrapolation of mass fluxes based on the frequency distribution of measured concentration and flow data, for discharges with absent data, suggests that the actual total mass flux for these contaminants could be up to 41% higher. The mass flux of Pb released from mines exceeds that of all currently permitted discharges (e.g. active industrial sites and wastewater treatment works) to surface waters across England and Wales, while those of As, Cd and Zn are of a similar magnitude. These data put into context the enduring legacy of historic mining on the water environment, highlighting its significance relative to more highly regulated polluting sites. Comparison of the figures with estimates of global trace metal flux suggests that the national total identified here is significant on a global scale.