Young guard cells function dynamically despite low mechanical anisotropy but gain efficiency during stomatal maturation in Arabidopsis thaliana.

Stomata are pores at the leaf surface that enable gas exchange and transpiration. The signaling pathways that regulate the differentiation of stomatal guard cells and the mechanisms of stomatal pore formation have been characterized in Arabidopsis thaliana. However, the process by which stomatal complexes develop after pore formation into fully mature complexes is poorly understood. We tracked the morphogenesis of young stomatal complexes over time to establish characteristic geometric milestones along the path of stomatal maturation. Using 3D-nanoindentation coupled with finite element modeling of young and mature stomata, we found that despite having thicker cell walls than young guard cells, mature guard cells are more energy efficient with respect to stomatal opening, potentially attributable to the increased mechanical anisotropy of their cell walls and smaller changes in turgor pressure between the closed and open states. Comparing geometric changes in young and mature guard cells of wild-type and cellulose-deficient plants revealed that although cellulose is required for normal stomatal maturation, mechanical anisotropy appears to be achieved by the collective influence of cellulose and additional wall components. Together, these data elucidate the dynamic geometric and biomechanical mechanisms underlying the development process of stomatal maturation.

Stomatal opening efficiency is controlled by cell wall organization in Arabidopsis thaliana.

Stomatal function in plants is regulated by the nanoscale architecture of the cell wall and turgor pressure, which together control stomatal pore size to facilitate gas exchange and photosynthesis. The mechanical properties of the cell wall and cell geometry are critical determinants of stomatal dynamics. However, the specific biomechanical functions of wall constituents, for example, cellulose and pectins, and their impact on the work required to open or close the stomatal pore are unclear. Here, we use nanoindentation in normal and lateral directions, computational modeling, and microscopic imaging of cells from the model plant Arabidopsis thaliana to investigate the precise influences of wall architecture and turgor pressure on stomatal biomechanics. This approach allows us to quantify and compare the unique anisotropic properties of guard cells with normal composition, lower cellulose content, or alterations in pectin molecular weight. Using these data to calculate the work required to open the stomata reveals that the wild type, with a circumferential-to-longitudinal modulus ratio of 3:1, is the most energy-efficient of those studied. In addition, the tested genotypes displayed similar changes in their pore size despite large differences in wall thickness and biomechanical properties. These findings imply that homeostasis in stomatal function is maintained in the face of varying wall compositions and biomechanics by tuning wall thickness.

Bone intrinsic material and compositional properties in postmenopausal women diagnosed with long-term Type-1 diabetes.

The incidence of diabetes mellitus and the associated complications are growing worldwide, affecting the patients' quality of life and exerting a considerable burden on health systems. Yet, the increase in fracture risk in type 1 diabetes (T1D) patients is not fully captured by bone mineral density (BMD), leading to the hypothesis that alterations in bone quality are responsible for the increased risk. Material/compositional properties are important aspects of bone quality, yet information on human bone material/compositional properties in T1D is rather sparse. The purpose of the present study is to measure both the intrinsic material behaviour by nanoindentation, and material compositional properties by Raman spectroscopy as a function of tissue age and microanatomical location (cement lines) in bone tissue from iliac crest biopsies from postmenopausal women diagnosed with long-term T1D (N = 8), and appropriate sex-, age-, BMD- and clinically-matched controls (postmenopausal women; N = 5). The results suggest elevation of advanced glycation endproducts (AGE) content in the T1D and show significant differences in mineral maturity / crystallinity (MMC) and glycosaminoglycan (GAG) content between the T1D and control groups. Furthermore, both hardness and modulus by nanoindentation are greater in T1D. These data suggest a significant deterioration of material strength properties (toughness) and compositional properties in T1D compared with controls.

Influence of tessellation morphology on ultrasonic scattering.

Material properties, such as hardness, yield strength, and ductility, depend on the microstructure of the material. If the microstructural organization can be quantified nondestructively, for example, with ultrasonic scattering techniques, then it may be possible to predict the mechanical performance of a component. Three-dimensional digital microstructures have been increasingly used to investigate the scattering of mechanical waves within a numerical framework. These synthetic microstructures can be generated using different tessellation algorithms that result in different grain shapes. In this study, the variation of ultrasonic scattering is calculated for microstructures of different morphologies for a nickel polycrystal. The ultrasonic properties are calculated for the Voronoi, Laguerre tessellations, and voxel-based synthetic microstructures created by DREAM.3D. The results show that the differences in the two-point statistics and ultrasonic attenuation for different morphologies become more significant at wider size distributions and higher frequencies.

Adapting to heatwave-induced seagrass loss: Prioritizing management areas through environmental sensitivity mapping.

Seagrass meadows support complex species assemblages and provide ecosystem services with a multitude of socio-economic benefits. However, they are sensitive to anthropogenic pressures such as coastal development, agricultural run-off, and overfishing. The increasing prevalence of marine heatwaves (MHWs) due to climate change poses an additional and growing threat. In this study, we apply the environmental sensitivity mapping approach MESA (Mapping Environmentally Sensitive Assets) to explore the potential consequences of MHWs on the ecosystem services that Posidonia oceanica provides to coastal communities. Under the intermediate climate change scenario Representative Concentration Pathway 4.5, Mediterranean marine heatwaves will be severe by 2050, and will very likely increase mortality of P. oceanica. However, the societal risk of seagrass loss is not evenly distributed across the Mediterranean. The spatial distribution of socio-economic implications of seagrass loss is highlighted through two case studies on seagrass-dependent fisheries and coastal hazards. Coastal communities in Tunisia and Libya show very high sensitivity to a loss of fisheries due to a combination of increasingly intense and frequent MHWs, coupled with high proportions of regional seagrass-dependent fisheries catch. The coastlines of Italy, Tunisia, and Cyprus are shown to potentially be highly sensitive to loss of seagrass due to high levels of coastal hazards, and seagrass meadows susceptible to MHW-induced degradation. These coastlines are likely to suffer from reduced coastal protection services provided by intact seagrass meadows. We demonstrate the implications of MHWs for ecosystem service provision to coastal communities in the Mediterranean and the need for policy instruments to help mitigate and adapt to its effect. We also highlight the potential for environmental sensitivity mapping to help support policymakers with rapid screening tools to prioritize resources more effectively to areas where in-depth local planning is needed.

A pilot study on the nanoscale properties of bone tissue near lacunae in fracturing women.

The goal of this study is to investigate the causes of osteoporosis-related skeletal fragility in postmenopausal women. We hypothesize that bone fragility in these individuals is largely due to mineral, and/or intrinsic material properties in the osteocyte lacunar/peri-lacunar regions of bone tissue. Innovative measurements with nanoscale resolution, including scanning electron microscope (SEM), an atomic force microscope that is integrated with infrared spectroscopy (AFM-IR), and nanoindentation, were used to characterize osteocyte lacunar and peri-lacunar properties in bone biopsies from fracturing (Cases) and matched (Age, BMD), non-fracturing (Controls) postmenopausal healthy women. In the peri-lacunar space, the nanoindentation results show that the modulus and hardness of the Controls are lower than the Cases. The AFM-IR results conclusively show that the mineral matrix, maturity (peak) (except in outer/far regions in Controls) were greater in Controls than in Cases. Furthermore, these results indicate that while mineral-to-matrix area ratio tend to be greater, the mineral maturity and crystallinity peak ratio "near" lacunae is greater than at regions "far" or more distance from lacunae in the Controls only. Due to the heterogeneity of bone structure, additional measurements are needed to provide more convincing evidence of altered lacunar characteristics and changes in the peri-lacunar bone as mechanisms related to postmenopausal women and fragility. Such findings would motivate new osteocyte-targeted treatments to reduce fragility fracture risks in these groups.

Cell twisting during desiccation reveals axial asymmetry in wall organization.

Plant cell size and shape are tuned to their function and specified primarily by cellulose microfibril (CMF) patterning of the cell wall. Arabidopsis thaliana leaf trichomes are unicellular structures that act as a physical defense to deter insect feeding. This highly polarized cell type employs a strongly anisotropic cellulose wall to extend and taper, generating sharply pointed branches. During elongation, the mechanisms by which shifts in fiber orientation generate cells with predictable sizes and shapes are unknown. Specifically, the axisymmetric growth of trichome branches is often thought to result from axisymmetric CMF patterning. Here, we analyzed the direction and degree of twist of branches after desiccation to reveal the presence of an asymmetric cell wall organization with a left-hand bias. CMF organization, quantified using computational modeling, suggests a limited reorientation of microfibrils during growth and a maximum branch length limited by the wall axial stiffness. The model provides a mechanism for CMF asymmetry, which occurs after the branch bending stiffness becomes low enough that ambient bending affects the principal stresses. After this stage, the CMF synthesis results in a constant bending stiffness for longer branches. The bending vibration natural frequencies of branches with respect to their length are also discussed.

Protocol for mapping the variability in cell wall mechanical bending behavior in living leaf pavement cells.

Mechanical properties, size and geometry of cells, and internal turgor pressure greatly influence cell morphogenesis. Computational models of cell growth require values for wall elastic modulus and turgor pressure, but very few experiments have been designed to validate the results using measurements that deform the entire thickness of the cell wall. New wall material is synthesized at the inner surface of the cell such that full-thickness deformations are needed to quantify relevant changes associated with cell development. Here, we present an integrated, experimental-computational approach to analyze quantitatively the variation of elastic bending behavior in the primary cell wall of living Arabidopsis (Arabidopsis thaliana) pavement cells and to measure turgor pressure within cells under different osmotic conditions. This approach used laser scanning confocal microscopy to measure the 3D geometry of single pavement cells and indentation experiments to probe the local mechanical responses across the periclinal wall. The experimental results were matched iteratively using a finite element model of the experiment to determine the local mechanical properties and turgor pressure. The resulting modulus distribution along the periclinal wall was nonuniform across the leaf cells studied. These results were consistent with the characteristics of plant cell walls which have a heterogeneous organization. The results and model allowed the magnitude and orientation of cell wall stress to be predicted quantitatively. The methods also serve as a reference for future work to analyze the morphogenetic behaviors of plant cells in terms of the heterogeneity and anisotropy of cell walls.

Real-time conversion of tissue-scale mechanical forces into an interdigitated growth pattern.

The leaf epidermis is a dynamic biomechanical shell that integrates growth across spatial scales to influence organ morphology. Pavement cells, the fundamental unit of this tissue, morph irreversibly into highly lobed cells that drive planar leaf expansion. Here, we define how tissue-scale cell wall tensile forces and the microtubule-cellulose synthase systems dictate the patterns of interdigitated growth in real time. A morphologically potent subset of cortical microtubules span the periclinal and anticlinal cell faces to pattern cellulose fibres that generate a patch of anisotropic wall. The subsequent local polarized growth is mechanically coupled to the adjacent cell via a pectin-rich middle lamella, and this drives lobe formation. Finite element pavement cell models revealed cell wall tensile stress as an upstream patterning element that links cell- and tissue-scale biomechanical parameters to interdigitated growth. Cell lobing in leaves is evolutionarily conserved, occurs in multiple cell types and is associated with important agronomic traits. Our general mechanistic models of lobe formation provide a foundation to analyse the cellular basis of leaf morphology and function.

PECTATE LYASE LIKE12 patterns the guard cell wall to coordinate turgor pressure and wall mechanics for proper stomatal function in Arabidopsis.

Plant cell deformations are driven by cell pressurization and mechanical constraints imposed by the nanoscale architecture of the cell wall, but how these factors are controlled at the genetic and molecular levels to achieve different types of cell deformation is unclear. Here, we used stomatal guard cells to investigate the influences of wall mechanics and turgor pressure on cell deformation and demonstrate that the expression of the pectin-modifying gene PECTATE LYASE LIKE12 (PLL12) is required for normal stomatal dynamics in Arabidopsis thaliana. Using nanoindentation and finite element modeling to simultaneously measure wall modulus and turgor pressure, we found that both values undergo dynamic changes during induced stomatal opening and closure. PLL12 is required for guard cells to maintain normal wall modulus and turgor pressure during stomatal responses to light and to tune the levels of calcium crosslinked pectin in guard cell walls. Guard cell-specific knockdown of PLL12 caused defects in stomatal responses and reduced leaf growth, which were associated with lower cell proliferation but normal cell expansion. Together, these results force us to revise our view of how wall-modifying genes modulate wall mechanics and cell pressurization to accomplish the dynamic cellular deformations that underlie stomatal function and tissue growth in plants.

Transverse-to-transverse diffuse ultrasonic double scattering.

Previously, a transverse-to-transverse single scattering model (T-T SSR) was developed for a pulse echo configuration, which may have limitations for strongly scattering materials. In this work, a transverse-to-transverse double scattering model (T-T DSR) is presented to model the transverse ultrasonic backscatter more accurately. First, the Wigner distribution of the transducer beam pattern is extended to a transverse wave. Next, the multiple scattering framework is followed to derive the transverse and longitudinal components of the second-order scattering. Then, a quasi-Monte Carlo (QMC) method is used with Graphics Processing Unit (GPU) acceleration to calculate numerical results of the final expression which contains a five-dimensional integral. The correlation length, the focal length of the transducer, and incident angle are used to investigate differences between the T-T DSR model and the T-T SSR model. Finally, a backscatter experiment is performed on two stainless steel specimens with different grain sizes to determine the respective correlation lengths. The results show that the T-T DSR model has better performance over the T-T SSR model for evaluating the grain size of these relatively strongly-scattering specimens.

Propagation of leaky Rayleigh waves along a curved fluid-solid interface.

A characteristic equation is derived for a leaky Rayleigh wave (LRW), propagating on a curved fluid-solid interface. The equations of motion for the curved solid and fluid are formulated using the constitutive equations of a homogenous isotropic curved solid and an inviscid fluid, respectively. The displacement potential functions are used to simplify the derivation. The interface conditions are used to ensure continuities of the mass, momentum, and energy across the interface. Then, with the consideration of the interface radius of the curvature, the characteristic equation for the LRW is established and solved numerically by Muller's method. One important outcome is that there is weaker directional dependence for the velocity of the LRWs on the radius of curvature in comparison with the Rayleigh waves at an air-solid interface. However, the numerical results show a strong directional dependence for the attenuation due to the LRW leakage on the complex curvatures. Moreover, a quantitative relation between the curvature and attenuation caused by the leakage for different materials is shown. The results are significant especially with respect to relevant future applications of ultrasonic testing.

Ultrasound in situ characterization of hybrid additively manufactured Ti6Al4V.

A major barrier for the full utilization of metal additive manufacturing (AM) technologies is quality control. Additionally, in situ real time nondestructive monitoring is desirable due to the typical high value and low volume of components manufactured with metal AM. Depending on the application, characteristics such as the geometrical accuracy, porosity, defect size and content, and material properties are quantities of interest for in situ nondestructive evaluation (NDE). In particular, functionally tailored components made with hybrid processing require quantitative NDE of their microstructure and elastic properties. Ultrasonic NDE is able to quantify these relevant characteristics. In this work, an ultrasonic measurement system is used to collect in situ real time measurements during the manufacturing of samples made with a hybrid process, which combines directed energy deposition with milling. In addition to quantifying ultrasonic properties, the measurements are used to gather insight on other geometry, material, and process effects. The results show the utility of ultrasound to evaluate relevant properties during manufacturing of a functionalized material domain, while providing perspective on additional material evolution information obtained from ultrasonic signals.

Ultrasonic mapping of hybrid additively manufactured 420 stainless steel.

Metal hybrid additive manufacturing (AM) processes are suitable to create complex structures that advance engineering performance. Hybrid AM can be used to create functionally graded materials for which the variation in microstructure and material properties across the domain is created through a synergized combination of fully-coupled manufacturing processes and/or energy sources. This expansion in the engineering design and manufacturing spaces presents challenges for nondestructive evaluation, including the assessment of the sensitivity of nondestructive measurements to functional gradients. To address this problem, linear ultrasound measurements are used to interrogate 420 stainless steel coupons from three manufacturing methods: wrought, AM, and hybrid AM (directed energy deposition + laser peening). Wave speed, attenuation, and diffuse backscatter results are compared with microhardness measurements along the build/axial direction of the coupons, while microstructure images are used for qualitative verification. The ultrasound measurements compare well with the destructive measurements without any substantial loss in resolution. Furthermore, ultrasonic methods are shown to be effective for identification of the gradient and cyclic nature of the elastic properties and microstructure on the hybrid AM coupon. These results highlight the potential of ultrasound as an efficient and accessible nondestructive characterization method for hybrid AM samples and inform further nondestructive evaluation decisions in AM.

Influence of microstructural grain-size distribution on ultrasonic scattering.

Ultrasonic attenuation and diffuse scattering result from the interaction of ultrasound with the microstructure of polycrystalline samples. Researchers are now using these effects to quantify mean grain size with good success and progress is being made with respect to more complex grain morphologies and macroscopic texture. However, theoretical models of such microstructures can become untenable because the scattering theory requires the two-point spatial statistics of the microstructure. For this reason, computational models of polycrystals are often considered for which grain spatial statistics can be calculated directly. In this study, the influence of grain-size distribution is examined using such an approach with three-dimensional (3D) realizations of polycrystalline materials. Representative material volumes are created using DREAM.3D with lognormal grain-size distributions, with fixed means but six different widths. These realizations are then used to calculate the relevant grain statistics which are then used to determine ultrasonic attenuation. The use of thirty realizations for each grain-size distribution allows the variation of the ultrasonic scattering to be quantified. The results show that the correlation between attenuation and distribution width can be modeled with a power law. Additionally, the frequency dependence of attenuation is shown to depend strongly on the distribution width. These results are expected to aid in the development of simplified models to quantify the grain-size distribution.

Mesophotic.org: a repository for scientific information on mesophotic ecosystems.

Mesophotic coral ecosystems (MCEs) and temperate mesophotic ecosystems (TMEs) occur at depths of roughly 30-150 m depth and are characterized by the presence of photosynthetic organisms despite reduced light availability. Exploration of these ecosystems dates back several decades, but our knowledge remained extremely limited until about a decade ago, when a renewed interest resulted in the establishment of a rapidly growing research community. Here, we present the 'mesophotic.org' database, a comprehensive and curated repository of scientific literature on mesophotic ecosystems. Through both manually curated and automatically extracted metadata, the repository facilitates rapid retrieval of available information about particular topics (e.g. taxa or geographic regions), exploration of spatial/temporal trends in research and identification of knowledge gaps. The repository can be queried to comprehensively obtain available data to address large-scale questions and guide future research directions. Overall, the 'mesophotic.org' repository provides an independent and open-source platform for the ever-growing research community working on MCEs and TMEs to collate and expedite our understanding of the occurrence, composition and functioning of these ecosystems. Database URL: http://mesophotic.org/.

Enhanced ultrasonic detection of near-surface flaws using transverse-wave backscatter.

Diffuse ultrasonic backscatter measurements have been shown to enhance the detection capability of sub-wavelength flaws when combined with extreme value statistics. However, for a normal-incidence immersion measurement, a "dead zone" created by the ring-down of the front-wall echo will hide near-surface flaws. In this article, a pulse-echo transverse wave backscatter measurement is used to detect near-surface flaws under high gain. The approach is validated using a magnesium specimen with side-drilled holes. The confidence bounds of the grain noise from this specimen are given by a transverse-to-transverse scattering model, which takes the grain size distribution and the hexagonal crystal symmetry into account. The upper bound is then treated as a time-dependent threshold for the C-scan. Experiments show that the developed method has good performance for detecting sub-wavelength, near-surface flaws, and can suppress both missed detections and false positives.

Ultrasonic wave propagation predictions for polycrystalline materials using three-dimensional synthetic microstructures: Phase velocity variations.

In most theoretical work related to effective properties of polycrystals, the media are assumed to be infinite with randomly oriented grains. Therefore, the bulk material has absolute isotropy because each direction includes an infinite number of grains with infinite possibilities for grain orientation. However, real samples will always include a finite number of grains such that the inspection volume will have some associated anisotropy. Thus, bounds on the bulk properties are expected for a given measurement. Here, the effect of the number of grains on the variations of elastic anisotropy is studied using synthetic polycrystals comprised of equiaxed cubic grains (17 volumes with 100 realizations each). Voigt, Reuss, and self-consistent techniques are used to derive the effective elastic modulus tensor. The standard deviation of the average elastic modulus is then quantified for several materials with varying degrees of single-crystal anisotropy and is shown to be inversely proportional to the square root of the number of grains. Finally, the Christoffel equation is used to study the relevant phase velocities. With appropriate normalization, a master curve is derived with respect to the finite sample size, which shows the expected variations of phase velocity for the longitudinal, fast shear, and slow shear modes.