Cretaceous chloranthoids: early prominence, extinct diversity and missing links.

BACKGROUND: The Chloranthaceae comprise four extant genera (Hedyosmum, Ascarina, Chloranthus and Sarcandra), all with simple flowers. Molecular phylogenetics indicates that the Chloranthaceae diverged very early in angiosperm evolution, although how they are related to eudicots, magnoliids, monocots and Ceratophyllum is uncertain. Fossil pollen similar to that of Ascarina and Hedyosmum has long been recognized in the Early Cretaceous, but over the last four decades evidence of extinct Chloranthaceae based on other types of fossils has expanded dramatically and contributes significantly to understanding the evolution of the family. SCOPE: Studies of fossils from the Cretaceous, especially mesofossils of Early Cretaceous age from Portugal and eastern North America, recognized diverse flowers, fruits, seeds, staminate inflorescences and stamens of extinct chloranthoids. These early chloranthoids include forms related to extant Hedyosmum and also to the Ascarina, Chloranthus and Sarcandra clade. In the Late Cretaceous there are several occurrences of distinctive fossil androecia related to extant Chloranthus. The rich and still expanding Cretaceous record of Chloranthaceae contrasts with a very sparse Cenozoic record, emphasizing that the four extant genera are likely to be relictual, although speciation within the genera might have occurred in relatively recent times. In this study, we describe three new genera of Early Cretaceous chloranthoids and summarize current knowledge on the extinct diversity of the group. CONCLUSIONS: The evolutionary lineage that includes extant Chloranthaceae is diverse and abundantly represented in Early Cretaceous mesofossil floras that provide some of the earliest evidence of angiosperm reproductive structures. Extinct chloranthoids, some of which are clearly in the Chloranthaceae crown group, fill some of the morphological gaps that currently separate the extant genera, help to illuminate how some of the unusual features of extant Chloranthaceae evolved and suggest that Chloranthaceae are of disproportionate importance for a more refined understanding of ecology and phylogeny of early angiosperm diversification.

A Collagen10a1 mutation disrupts cell polarity in a medaka model for metaphyseal chondrodysplasia type Schmid.

Heterozygous mutations in COL10A1 lead to metaphyseal chondrodysplasia type Schmid (MCDS), a skeletal disorder characterized by epiphyseal abnormalities. Prior analysis revealed impaired trimerization and intracellular retention of mutant collagen type X alpha 1 chains as cause for elevated endoplasmic reticulum (ER) stress. However, how ER stress translates into structural defects remained unclear. We generated a medaka (Oryzias latipes) MCDS model harboring a 5 base pair deletion in col10a1, which led to a frameshift and disruption of 11 amino acids in the conserved trimerization domain. col10a1Δ633a heterozygotes recapitulated key features of MCDS and revealed early cell polarity defects as cause for dysregulated matrix secretion and deformed skeletal structures. Carbamazepine, an ER stress-reducing drug, rescued this polarity impairment and alleviated skeletal defects in col10a1Δ633a heterozygotes. Our data imply cell polarity dysregulation as a potential contributor to MCDS and suggest the col10a1Δ633a medaka mutant as an attractive MCDS animal model for drug screening.

Heitt Mjölnir: a heated miniature triaxial apparatus for 4D synchrotron microtomography.

Third- and fourth-generation synchrotron light sources with high fluxes and beam energies enable the use of innovative X-ray translucent experimental apparatus. These experimental devices access geologically relevant conditions whilst enabling in situ characterization using the spatial and temporal resolutions accessible at imaging beamlines. Here, Heitt Mjölnir is introduced, a heated miniature triaxial rig based on the design of Mjölnir, but covering a wider temperature range and larger sample volume at similar pressure capacities. This device is designed to investigate coupled thermal, chemical, hydraulic and mechanical processes from grain to centimetre scales using cylindrical samples of 10 mm × 20 mm (diameter × length). Heitt Mjölnir can simultaneously reach confining (hydraulic) pressures of 30 MPa and 500 MPa of axial stress with independently controlled sample pore fluid pressure < 30 MPa. This internally heated apparatus operates to temperatures up to 573 K with a minimal vertical thermal gradient in the sample of <0.3 K mm-1. This new apparatus has been deployed in operando studies at the TOMCAT (Swiss Light Source), I12 JEEP (Diamond Light Source) and PSICHÉ (Synchrotron SOLEIL) beamlines for 4D X-ray microtomography with scan intervals of a few minutes. Heitt Mjölnir is portable and modular, allowing a wide range of 4D characterizations of low-grade metamorphism and deformational processes. It enables spatially and temporally resolved fluid-rock interaction studies at conditions of crustal reservoirs and is suitable for characterization of material properties in geothermal, carbonation or subsurface gas storage applications. Technical drawings and an operation guide are included in this publication.

Harmonizing sound and light: X-ray imaging unveils acoustic signatures of stochastic inter-regime instabilities during laser melting.

Laser powder bed fusion (LPBF) is a metal additive manufacturing technique involving complex interplays between vapor, liquid, and solid phases. Despite LPBF's advantageous capabilities compared to conventional manufacturing methods, the underlying physical phenomena can result in inter-regime instabilities followed by transitions between conduction and keyhole melting regimes - leading to defects. We investigate these issues through operando synchrotron X-ray imaging synchronized with acoustic emission recording, during the remelting processes of LPBF-produced thin walls, monitoring regime changes occurring under constant laser processing parameters. The collected data show an increment in acoustic signal amplitude when switching from conduction to keyhole regime, which we correlate to changes in laser absorptivity. Moreover, a full correlation between X-ray imaging and the acoustic signals permits the design of a simple filtering algorithm to predict the melting regimes. As a result, conduction, stable keyhole, and unstable keyhole regimes are identified with a time resolution of 100 µs, even under rapid transitions, providing a straightforward method to accurately detect undesired processing regimes without the use of artificial intelligence.

On the water transport mechanism through the microporous layers of operando polymer electrolyte fuel cells probed directly by X-ray tomographic microscopy.

Product water transport via the microporous layer (MPL) and gas diffusion layer (GDL) substrate during polymer electrolyte fuel cell (PEFC) operation was directly and quantitatively observed by X-ray tomographic microscopy (XTM). The liquid water distribution in two types of MPLs with different pore size distributions (PSDs) was characterized as a function of the inlet gas relative humidity (RH) and current density under humid operating conditions at 45 °C. During the first minute of PEFC operation, liquid water mainly accumulated at the catalyst layer (CL)/MPL interface and in the GDL substrate close to the flow fields. Furthermore, under all tested conditions, saturation in the MPL was low (<25%), whereas under the rib, the saturation in the GDL was up to ca. 70%. Based on these XTM results, it is confirmed that in the high porosity MPLs, vapor transport was non-negligible even at high humidity conditions. Therefore, on top of the widely discussed MPL pore size and its distribution, it is proposed that the lower thermal conductivity from the high porosity of MPLs can also be a main cause of promoted vapor transport, reducing water saturation near the CL.

Structural formation during bread baking in a combined microwave-convective oven determined by sub-second in-situ synchrotron X-ray microtomography.

A new concept has been developed for characterizing the real-time evolution of the three-dimensional pore and lamella microstructure of bread during baking using synchrotron X-ray microtomography (SRµCT). A commercial, combined microwave-convective oven was modified and installed at the TOMCAT synchrotron tomography beamline at the Swiss Light Source (SLS), to capture the 3D dough-to-bread structural development in-situ at the micrometer scale with an acquisition time of 400 ms. This allowed characterization and quantitative comparison of three baking technologies: (1) convective heating, (2) microwave heating, and (3) a combination of convective and microwave heating. A workflow for automatic batchwise image processing and analysis of 3D bread structures (1530 analyzed volumes in total) was established for porosity, individual pore volume, elongation, coordination number and local wall thickness, which allowed for evaluation of the impact of baking technology on the bread structure evolution. The results showed that the porosity, mean pore volume and mean coordination number increase with time and that the mean local cell wall thickness decreases with time. Small and more isolated pores are connecting with larger and already more connected pores as function of time. Clear dependencies are established during the whole baking process between the mean pore volume and porosity, and between the mean local wall thickness and the mean coordination number. This technique opens new opportunities for understanding the mechanisms governing the structural changes during baking and discern the parameters controlling the final bread quality.

Nanoplastics causes extensive congenital malformations during embryonic development by passively targeting neural crest cells.

Nanomaterials are widespread in the human environment as pollutants, and are being actively developed for use in human medicine. We have investigated how the size and dose of polystyrene nanoparticles affects malformations in chicken embryos, and have characterized the mechanisms by which they interfere with normal development. We find that nanoplastics can cross the embryonic gut wall. When injected into the vitelline vein, nanoplastics become distributed in the circulation to multiple organs. We find that the exposure of embryos to polystyrene nanoparticles produces malformations that are far more serious and extensive than has been previously reported. These malformations include major congenital heart defects that impair cardiac function. We show that the mechanism of toxicity is the selective binding of polystyrene nanoplastics nanoparticles to neural crest cells, leading to the death and impaired migration of those cells. Consistent with our new model, most of the malformations seen in this study are in organs that depend for their normal development on neural crest cells. These results are a matter of concern given the large and growing burden of nanoplastics in the environment. Our findings suggest that nanoplastics may pose a health risk to the developing embryo.

Investigating microstructure evolution of lithium metal during plating and stripping via operando X-ray tomographic microscopy.

Efficient lithium metal stripping and plating operation capable of maintaining electronic and ionic conductivity is crucial to develop safe lithium metal batteries. However, monitoring lithium metal microstructure evolution during cell cycling is challenging. Here, we report the development of an operando synchrotron X-ray tomographic microscopy method capable of probing in real-time the formation, growth, and dissolution of Li microstructures during the cycling of a Li||Cu cell containing a standard non-aqueous liquid electrolyte solution. The analyses of the operando X-ray tomographic microscopy measurements enable tracking the evolution of deposited Li metal as a function of time and applied current density and distinguishing the formation of electrochemically inactive Li from the active bulk of Li microstructures. Furthermore, in-depth analyses of the Li microstructures shed some light on the structural connectivity of deposited Li at different current densities as well as the formation mechanism of fast-growing fractal Li microstructures, which are ultimately responsible for cell failure.

Exploration of the X-ray Dark-Field Signal in Mineral Building Materials.

Mineral building materials suffer from weathering processes such as salt efflorescence, freeze-thaw cycling, and microbial colonization. All of these processes are linked to water (liquid and vapor) in the pore space. The degree of damage following these processes is heavily influenced by pore space properties such as porosity, pore size distribution, and pore connectivity. X-ray computed micro-tomography (µCT) has proven to be a valuable tool to non-destructively investigate the pore space of stone samples in 3D. However, a trade-off between the resolution and field-of-view often impedes reliable conclusions on the material's properties. X-ray dark-field imaging (DFI) is based on the scattering of X-rays by sub-voxel-sized features, and as such, provides information on the sample complementary to that obtained using conventional µCT. In this manuscript, we apply X-ray dark-field tomography for the first time on four mineral building materials (quartzite, fired clay brick, fired clay roof tile, and carbonated mineral building material), and investigate which information the dark-field signal entails on the sub-resolution space of the sample. Dark-field tomography at multiple length scale sensitivities was performed at the TOMCAT beamline of the Swiss Light Source (Villigen, Switzerland) using a Talbot grating interferometer. The complementary information of the dark-field modality is most clear in the fired clay brick and roof tile; quartz grains that are almost indistinguishable in the conventional µCT scan are clearly visible in the dark-field owing to their low dark-field signal (homogenous sub-voxel structure), whereas the microporous bulk mass has a high dark-field signal. Large (resolved) pores on the other hand, which are clearly visible in the absorption dataset, are almost invisible in the dark-field modality because they are overprinted with dark-field signal originating from the bulk mass. The experiments also showed how the dark-field signal from a feature depends on the length scale sensitivity, which is set by moving the sample with respect to the grating interferometer.

Saccorhytus is an early ecdysozoan and not the earliest deuterostome.

The early history of deuterostomes, the group composed of the chordates, echinoderms and hemichordates1, is still controversial, not least because of a paucity of stem representatives of these clades2-5. The early Cambrian microscopic animal Saccorhytus coronarius was interpreted as an early deuterostome on the basis of purported pharyngeal openings, providing evidence for a meiofaunal ancestry6 and an explanation for the temporal mismatch between palaeontological and molecular clock timescales of animal evolution6-8. Here we report new material of S. coronarius, which is reconstructed as a millimetric and ellipsoidal meiobenthic animal with spinose armour and a terminal mouth but no anus. Purported pharyngeal openings in support of the deuterostome hypothesis6 are shown to be taphonomic artefacts. Phylogenetic analyses indicate that S. coronarius belongs to total-group Ecdysozoa, expanding the morphological disparity and ecological diversity of early Cambrian ecdysozoans.

Deep learning based classification of dynamic processes in time-resolved X-ray tomographic microscopy.

Time-resolved X-ray tomographic microscopy is an invaluable technique to investigate dynamic processes in 3D for extended time periods. Because of the limited signal-to-noise ratio caused by the short exposure times and sparse angular sampling frequency, obtaining quantitative information through post-processing remains challenging and requires intensive manual labor. This severely limits the accessible experimental parameter space and so, prevents fully exploiting the capabilities of the dedicated time-resolved X-ray tomographic stations. Though automatic approaches, often exploiting iterative reconstruction methods, are currently being developed, the required computational costs typically remain high. Here, we propose a highly efficient reconstruction and classification pipeline (SIRT-FBP-MS-D-DIFF) that combines an algebraic filter approximation and machine learning to significantly reduce the computational time. The dynamic features are reconstructed by standard filtered back-projection with an algebraic filter to approximate iterative reconstruction quality in a computationally efficient manner. The raw reconstructions are post-processed with a trained convolutional neural network to extract the dynamic features from the low signal-to-noise ratio reconstructions in a fully automatic manner. The capabilities of the proposed pipeline are demonstrated on three different dynamic fuel cell datasets, one exploited for training and two for testing without network retraining. The proposed approach enables automatic processing of several hundreds of datasets in a single day on a single GPU node readily available at most institutions, so extending the possibilities in future dynamic X-ray tomographic investigations.

Fast acquisition protocol for X-ray scattering tensor tomography.

Microstructural information over an entire sample is important to understand the macroscopic behaviour of materials. X-ray scattering tensor tomography facilitates the investigation of the microstructural organisation in statistically large sample volumes. However, established acquisition protocols based on scanning small-angle X-ray scattering and X-ray grating interferometry inherently require long scan times even with highly brilliant X-ray sources. Recent developments in X-ray diffractive optics towards circular pattern arrays enable fast single-shot acquisition of the sample scattering properties with 2D omnidirectional sensitivity. X-ray scattering tensor tomography with the use of this circular grating array has been demonstrated. We propose here simple yet inherently rapid acquisition protocols for X-ray scattering tensor tomography leveraging on these new optical elements. Results from both simulation and experimental data, supported by a null space analysis, suggest that the proposed acquisition protocols are not only rapid but also corroborate that sufficient information for the accurate volumetric reconstruction of the scattering properties is provided. The proposed acquisition protocols will build the basis for rapid inspection and/or time-resolved tensor tomography of the microstructural organisation over an extended field of view.

Tunable X-ray dark-field imaging for sub-resolution feature size quantification in porous media.

X-ray computed micro-tomography typically involves a trade-off between sample size and resolution, complicating the study at a micrometer scale of representative volumes of materials with broad feature size distributions (e.g. natural stones). X-ray dark-field tomography exploits scattering to probe sub-resolution features, promising to overcome this trade-off. In this work, we present a quantification method for sub-resolution feature sizes using dark-field tomograms obtained by tuning the autocorrelation length of a Talbot grating interferometer. Alumina particles with different nominal pore sizes (50 nm and 150 nm) were mixed and imaged at the TOMCAT beamline of the SLS synchrotron (PSI) at eighteen correlation lengths, covering the pore size range. The different particles cannot be distinguished by traditional absorption µCT due to their very similar density and the pores being unresolved at typical image resolutions. Nevertheless, by exploiting the scattering behavior of the samples, the proposed analysis method allowed to quantify the nominal pore sizes of individual particles. The robustness of this quantification was proven by reproducing the experiment with solid samples of alumina, and alumina particles that were kept separated. Our findings demonstrate the possibility to calibrate dark-field image analysis to quantify sub-resolution feature sizes, allowing multi-scale analyses of heterogeneous materials without subsampling.

Molecular mechanisms and hormonal regulation underpinning morphological dormancy: a case study using Apium graveolens (Apiaceae).

Underdeveloped (small) embryos embedded in abundant endosperm tissue, and thus having morphological dormancy (MD) or morphophysiological dormancy (MPD), are considered to be the ancestral state in seed dormancy evolution. This trait is retained in the Apiaceae family, which provides excellent model systems for investigating the underpinning mechanisms. We investigated Apium graveolens (celery) MD by combined innovative imaging and embryo growth assays with the quantification of hormone metabolism, as well as the analysis of hormone and cell-wall related gene expression. The integrated experimental results demonstrated that embryo growth occurred inside imbibed celery fruits in association with endosperm degradation, and that a critical embryo size was required for radicle emergence. The regulation of these processes depends on gene expression leading to gibberellin and indole-3-acetic acid (IAA) production by the embryo and on crosstalk between the fruit compartments. ABA degradation associated with distinct spatiotemporal patterns in ABA sensitivity control embryo growth, endosperm breakdown and radicle emergence. This complex interaction between gibberellins, IAA and ABA metabolism, and changes in the tissue-specific sensitivities to these hormones is distinct from non-MD seeds. We conclude that the embryo growth to reach the critical size and the associated endosperm breakdown inside MD fruits constitute a unique germination programme.

X-ray nanotomography and electron backscatter diffraction demonstrate the crystalline, heterogeneous and impermeable nature of conodont white matter.

Conodont elements, microfossil remains of extinct primitive vertebrates, are commonly exploited as mineral archives of ocean chemistry, yielding fundamental insights into the palaeotemperature and chemical composition of past oceans. Geochemical assays have been traditionally focused on the so-called lamellar and white matter crown tissues; however, the porosity and crystallographic nature of the white matter and its inferred permeability are disputed, raising concerns over its suitability as a geochemical archive. Here, we constrain the characteristics of this tissue and address conflicting interpretations using ptychographic X-ray-computed tomography (PXCT), pore network analysis, synchrotron radiation X-ray tomographic microscopy (srXTM) and electron back-scatter diffraction (EBSD). PXCT and pore network analyses based on these data reveal that while white matter is extremely porous, the pores are unconnected, rendering this tissue closed to postmortem fluid percolation. EBSD analyses demonstrate that white matter is crystalline and comprised of a single crystal typically tens of micrometres in dimensions. Combined with evidence that conodont elements grow episodically, these data suggest that white matter, which comprises the denticles of conodont elements, grows syntactically, indicating that individual crystals are time heterogeneous. Together these data provide support for the interpretation of conodont white matter as a closed geochemical system and, therefore, its utility of the conodont fossil record as a historical archive of Palaeozoic and Early Mesozoic ocean chemistry.

Operando Liquid Pressure Determination in Polymer Electrolyte Fuel Cells.

Extending the operating range of fuel cells to higher current densities is limited by the ability of the cell to remove the water produced by the electrochemical reaction, avoiding flooding of the gas diffusion layers. It is therefore of great interest to understand the complex and dynamic mechanisms of water cluster formation in an operando fuel cell setting as this can elucidate necessary changes to the gas diffusion layer properties with the goal of minimizing the number, size, and instability of the water clusters formed. In this study, we investigate the cluster formation process using X-ray tomographic microscopy at 1 Hz frequency combined with interfacial curvature analysis and volume-of-fluid simulations to assess the pressure evolution in the water phase. This made it possible to observe the increase in capillary pressure when the advancing water front had to overcome a throat between two neighboring pores and the nuanced interactions of volume and pressure evolution during the droplet formation and its feeding path instability. A 2 kPa higher breakthrough pressure compared to static ex situ capillary pressure versus saturation evaluations was observed, which suggests a rethinking of the dynamic liquid water invasion process in polymer electrolyte fuel cell gas diffusion layers.

Deep denoising for multi-dimensional synchrotron X-ray tomography without high-quality reference data.

Synchrotron X-ray tomography enables the examination of the internal structure of materials at submicron spatial resolution and subsecond temporal resolution. Unavoidable experimental constraints can impose dose and time limits on the measurements, introducing noise in the reconstructed images. Convolutional neural networks (CNNs) have emerged as a powerful tool to remove noise from reconstructed images. However, their training typically requires collecting a dataset of paired noisy and high-quality measurements, which is a major obstacle to their use in practice. To circumvent this problem, methods for CNN-based denoising have recently been proposed that require no separate training data beyond the already available noisy reconstructions. Among these, the Noise2Inverse method is specifically designed for tomography and related inverse problems. To date, applications of Noise2Inverse have only taken into account 2D spatial information. In this paper, we expand the application of Noise2Inverse in space, time, and spectrum-like domains. This development enhances applications to static and dynamic micro-tomography as well as X-ray diffraction tomography. Results on real-world datasets establish that Noise2Inverse is capable of accurate denoising and enables a substantial reduction in acquisition time while maintaining image quality.

A tale of two morphs: developmental patterns and mechanisms of seed coat differentiation in the dimorphic diaspore model Aethionema arabicum (Brassicaceae).

The developmental transition from a fertilized ovule to a dispersed diaspore (seed or fruit) involves complex differentiation processes of the ovule's integuments leading to the diversity in mature seed coat structures in angiosperms. In this study, comparative imaging and transcriptome analysis were combined to investigate the morph-specific developmental differences during outer seed coat differentiation and mucilage production in Aethionema arabicum, the Brassicaceae model for diaspore dimorphism. One of the intriguing adaptations of this species is the production and dispersal of morphologically distinct, mucilaginous and non-mucilaginous diaspores from the same plant (dimorphism). The dehiscent fruit morph programme producing multiple mucilaginous seed diaspores was used as the default trait combination, similar to Arabidopsis thaliana, and was compared with the indehiscent fruit morph programme leading to non-mucilaginous diaspores. Synchrotron-based radiation X-ray tomographic microscopy revealed a co-ordinated framework of morph-specific early changes in internal anatomy of developing A. arabicum gynoecia including seed abortion in the indehiscent programme and mucilage production by the mucilaginous seed coat. The associated comparative analysis of the gene expression patterns revealed that the unique seed coat dimorphism of Ae. arabicum provides an excellent model system for comparative study of the control of epidermal cell differentiation and mucilage biosynthesis by the mucilage transcription factor cascade and their downstream cell wall and mucilage remodelling genes. Elucidating the underlying molecular framework of the dimorphic diaspore syndrome is key to understanding differential regulation of bet-hedging survival strategies in challenging environments, timely in the face of global climatic change.

Acanthodian dental development and the origin of gnathostome dentitions.

Chondrichthyan dentitions are conventionally interpreted to reflect the ancestral gnathostome condition but interpretations of osteichthyan dental evolution in this light have proved unsuccessful, perhaps because chondrichthyan dentitions are equally specialized, or else evolved independently. Ischnacanthid acanthodians are stem-Chondrichthyes; as phylogenetic intermediates of osteichthyans and crown-chondrichthyans, the nature of their enigmatic dentition may inform homology and the ancestral gnathostome condition. Here we show that ischnacanthid marginal dentitions were statodont, composed of multicuspidate teeth added in distally diverging rows and through proximal superpositional replacement, while their symphyseal tooth whorls are comparable to chondrichthyan and osteichthyan counterparts. Ancestral state estimation indicates the presence of oral tubercles on the jaws of the gnathostome crown-ancestor; tooth whorls or tooth rows evolved independently in placoderms, osteichthyans, ischnacanthids, other acanthodians and crown-chondrichthyans. Crown-chondrichthyan dentitions are derived relative to the gnathostome crown-ancestor, which possessed a simple dentition and lacked a permanent dental lamina, which evolved independently in Chondrichthyes and Osteichthyes.