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.

Design and Study of a Reflector-Separated Light Dispersion-Compensated 3D Microscopy System.

The secondary-phase grating-based tomographic microscopy system, which is widely used in the biological and life sciences, can observe all the sample multilayer image information simultaneously because it has multifocal points. However, chromatic aberration exists in the grating diffraction, which seriously affects the observation of the image. To correct the chromatic aberration of the tomographic microscope system, this paper proposes a system that adopts blazed gratings and angle-variable reflectors as chromatic aberration correction devices according to the principle of dispersion compensation and Fourier phase-shift theory. A reflector-separated light dispersion-compensated 3D microscopy system is presented to achieve chromatic aberration correction while solving the problem of multilayer image overlap. The theoretical verification and optical design of the system were completed using ZEMAX software. The results show that the proposed system reduced the chromatic aberration of ordinary tomographic microscopy systems by more than 90%, retaining more wavelengths of light information. In addition, the system had a relatively wide range in the color difference compensation element installation position, reducing the difficulty of dispersion compensation element installation. Overall, the results indicate that the proposed system is effective in reducing chromatic aberration in grating diffraction.

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.

High-numerical-aperture macroscope optics for time-resolved experiments.

A novel high-quality custom-made macroscope optics, dedicated to high-resolution time-resolved X-ray tomographic microscopy at the TOMCAT beamline at the Swiss Light Source (Paul Scherrer Institut, Switzerland), is introduced. The macroscope offers 4× magnification, has a very high numerical aperture of 0.35 and it is modular and highly flexible. It can be mounted both in a horizontal and vertical configuration, enabling imaging of tall samples close to the scintillator, to avoid edge-enhancement artefacts. The macroscope performance was characterized and compared with two existing in-house imaging setups, one dedicated to high spatial and one to high temporal resolution. The novel macroscope shows superior performance for both imaging settings compared with the previous systems. For the time-resolved setup, the macroscope is 4 times more efficient than the previous system and, at the same time, the spatial resolution is also increased by a factor of 6. For the high-spatial-resolution setup, the macroscope is up to 8.5 times more efficient with a moderate spatial resolution improvement (factor of 1.5). This high efficiency, increased spatial resolution and very high image quality offered by the novel macroscope optics will make 10-20 Hz high-resolution tomographic studies routinely possible, unlocking unprecedented possibilities for the tomographic investigations of dynamic processes and radiation-sensitive samples.

Tanispermum, a new genus of hemi-orthotropous to hemi-anatropous angiosperm seeds from the Early Cretaceous of eastern North America.

PREMISE OF THE STUDY: Exotestal seeds with characters that indicate relationship to extant Austrobaileyales and Nymphaeales are abundant in Early Cretaceous sediments from Portugal and eastern North America, but their variety and unique features provide evidence of extensive extinct diversity among early angiosperms. METHODS: The fossils were extracted from Early Cretaceous sediments from Virginia and Maryland, United States, by sieving in water. After cleaning with HF, HCl and water, they were examined using SEM and SRXTM and compared to seeds of extant and fossil angiosperms. KEY RESULTS: A new genus, Tanispermum gen. nov., with four species (T. hopewellense sp. nov., T. marylandense sp. nov., T. drewriense sp. nov., and T. antiquum sp. nov.) is recognized. Relationship with extant Austrobaileyales and Nymphaeales is indicated by an exotesta of tall palisade sclerenchyma with undulate anticlinal walls forming a jigsaw puzzle-like surface pattern. However, seeds of Tanispermum differ from those of Austrobaileyales and Nymphaeales in their hemi-orthotropous-hemi-anatropous organization. Attempts to place Tanispermum in a phylogenetic context confront a variety of problems, including lack of information on other parts of these extinct plants. CONCLUSIONS: The discovery highlights the extent to which the morphology of extant angiosperms is not representative of the diversity that once existed among early-diverging members of the group. The relictual nature of extant taxa near the base of the angiosperm tree greatly complicates the reconstruction of ancestral character states and emphasizes the need for focused paleobotanical studies to elucidate the extinct diversity that existed early in angiosperm evolution.

Effects of nintedanib on the microvascular architecture in a lung fibrosis model.

Nintedanib, a tyrosine kinase inhibitor approved for the treatment of idiopathic pulmonary fibrosis, has anti-fibrotic, anti-inflammatory, and anti-angiogenic activity. We explored the impact of nintedanib on microvascular architecture in a pulmonary fibrosis model. Lung fibrosis was induced in C57Bl/6 mice by intratracheal bleomycin (0.5 mg/kg). Nintedanib was started after the onset of lung pathology (50 mg/kg twice daily, orally). Micro-computed tomography was performed via volumetric assessment. Static lung compliance and forced vital capacity were determined by invasive measurements. Mice were subjected to bronchoalveolar lavage and histologic analyses, or perfused with a casting resin. Microvascular corrosion casts were imaged by scanning electron microscopy and synchrotron radiation tomographic microscopy, and quantified morphometrically. Bleomycin administration resulted in a significant increase in higher-density areas in the lungs detected by micro-computed tomography, which was significantly attenuated by nintedanib. Nintedanib significantly reduced lung fibrosis and vascular proliferation, normalized the distorted microvascular architecture, and was associated with a trend toward improvement in lung function and inflammation. Nintedanib resulted in a prominent improvement in pulmonary microvascular architecture, which outperformed the effect of nintedanib on lung function and inflammation. These findings uncover a potential new mode of action of nintedanib that may contribute to its efficacy in idiopathic pulmonary fibrosis.

Fast iterative reconstruction of data in full interior tomography.

This paper introduces two novel strategies for iterative reconstruction of full interior tomography (FINT) data, i.e. when the field of view is entirely inside the object support and knowledge of the object support itself or the attenuation coefficients inside specific regions of interest are not available. The first approach is based on data edge-padding. The second technique creates an intermediate virtual sinogram, which is, then, reconstructed by a standard iterative algorithm. Both strategies are validated in the framework of the alternate direction method of multipliers plug-and-play with gridding projectors that provide a speed-up of three orders of magnitude with respect to standard operators implemented in real space. The proposed methods are benchmarked on synchrotron-based X-ray tomographic microscopy datasets of mouse lung alveoli. Compared with analytical techniques, the proposed methods substantially improve the reconstruction quality for FINT underconstrained datasets, facilitating subsequent post-processing steps.

GigaFRoST: the gigabit fast readout system for tomography.

Owing to recent developments in CMOS technology, it is now possible to exploit tomographic microscopy at third-generation synchrotron facilities with unprecedented speeds. Despite this rapid technical progress, one crucial limitation for the investigation of realistic dynamic systems has remained: a generally short total acquisition time at high frame rates due to the limited internal memory of available detectors. To address and solve this shortcoming, a new detection and readout system, coined GigaFRoST, has been developed based on a commercial CMOS sensor, acquiring and streaming data continuously at 7.7 GB s-1 directly to a dedicated backend server. This architecture allows for dynamic data pre-processing as well as data reduction, an increasingly indispensable step considering the vast amounts of data acquired in typical fast tomographic experiments at synchrotron beamlines (up to several tens of TByte per day of raw data).

Automated segmentation of wood fibres in micro-CT images of paper.

A novel algorithm has been developed and validated to isolate individual papermaking fibres in micro-computed tomographic images of paper handsheets as a first step to characterize the structure of the paper. The three-step fibre segmentation algorithm segments the papermaking fibres by (i) tracking the hollow inside the fibres via a modified connected component methodology, (ii) extracting the fibre walls using a distance transform and (iii) labelling the fibres through collapsed sections by a final refinement step. Furthermore, postprocessing algorithms have been developed to calculate the length and coarseness of the segmented fibres. The fibre segmentation algorithm is the first ever reported method for the automated segmentation of the tortuous three-dimensional morphology of papermaking fibres within microstructural images of paper handsheets. The method is not limited to papermaking fibres, but can be applied to any material consisting of tortuous and hollow fibres.

Visualization of water drying in porous materials by X-ray phase contrast imaging.

We present in this study results from X-ray tomographic microscopy with synchrotron radiation performed both in attenuation and phase contrast modes on a limestone sample during two stages of water drying. No contrast agent was used in order to increase the X-ray attenuation by water. We show that only by using the phase contrast mode it is possible to achieve enough water content change resolution to investigate the drying process at the pore-scale. We performed 3D image analysis of the time-differential phase contrast tomogram. We show by the results of such analysis that it is possible to obtain a reliable characterization of the spatial redistribution of water in the resolved pore system in agreement with what expected from the theory of drying in porous media and from measurements performed with other approaches. We thus show the potential of X-ray phase contrast imaging for pore-scale investigations of reactive water transport processes which cannot be imaged by adding a contrast agent for exploiting the standard attenuation contrast imaging mode.

Seed morphology and anatomy and its utility in recognizing subfamilies and tribes of Zingiberaceae.

PREMISE OF THE STUDY: Recent phylogenetic analyses based on molecular data suggested that the monocot family Zingiberaceae be separated into four subfamilies and four tribes. Robust morphological characters to support these clades are lacking. Seeds were analyzed in a phylogenetic context to test independently the circumscription of clades and to better understand evolution of seed characters within Zingiberaceae. METHODS: Seventy-five species from three of the four subfamilies were analyzed using synchrotron based x-ray tomographic microscopy (SRXTM) and scored for 39 morphoanatomical characters. KEY RESULTS: Zingiberaceae seeds are some of the most structurally complex seeds in angiosperms. No single seed character was found to distinguish each subfamily, but combinations of characters were found to differentiate between the subfamilies. Recognition of the tribes based on seeds was possible for Globbeae, but not for Alpinieae, Riedelieae, or Zingibereae, due to considerable variation. CONCLUSIONS: SRXTM is an excellent, nondestructive tool to capture morphoanatomical variation of seeds and allows for the study of taxa with limited material available. Alpinioideae, Siphonochiloideae, Tamijioideae, and Zingiberoideae are well supported based on both molecular and morphological data, including multiple seed characters. Globbeae are well supported as a distinctive tribe within the Zingiberoideae, but no other tribe could be differentiated using seeds due to considerable homoplasy when compared with currently accepted relationships based on molecular data. Novel seed characters suggest tribal affinities for two currently unplaced Zingiberaceae taxa: Siliquamomum may be related to Riedelieae and Monolophus to Zingibereae, but further work is needed before formal revision of the family.

Quantifying phosphoric acid in high-temperature polymer electrolyte fuel cell components by X-ray tomographic microscopy.

Synchrotron-based X-ray tomographic microscopy is investigated for imaging the local distribution and concentration of phosphoric acid in high-temperature polymer electrolyte fuel cells. Phosphoric acid fills the pores of the macro- and microporous fuel cell components. Its concentration in the fuel cell varies over a wide range (40-100 wt% H3PO4). This renders the quantification and concentration determination challenging. The problem is solved by using propagation-based phase contrast imaging and a referencing method. Fuel cell components with known acid concentrations were used to correlate greyscale values and acid concentrations. Thus calibration curves were established for the gas diffusion layer, catalyst layer and membrane in a non-operating fuel cell. The non-destructive imaging methodology was verified by comparing image-based values for acid content and concentration in the gas diffusion layer with those from chemical analysis.

Airspace Diameter Map-A Quantitative Measurement of All Pulmonary Airspaces to Characterize Structural Lung Diseases.

(1) Background: Stereological estimations significantly contributed to our understanding of lung anatomy and physiology. Taking stereology fully 3-dimensional facilitates the estimation of novel parameters. (2) Methods: We developed a protocol for the analysis of all airspaces of an entire lung. It includes (i) high-resolution synchrotron radiation-based X-ray tomographic microscopy, (ii) image segmentation using the free machine-learning tool Ilastik and ImageJ, and (iii) calculation of the airspace diameter distribution using a diameter map function. To evaluate the new pipeline, lungs from adult mice with cystic fibrosis (CF)-like lung disease (ßENaC-transgenic mice) or mice with elastase-induced emphysema were compared to healthy controls. (3) Results: We were able to show the distribution of airspace diameters throughout the entire lung, as well as separately for the conducting airways and the gas exchange area. In the pathobiological context, we observed an irregular widening of parenchymal airspaces in mice with CF-like lung disease and elastase-induced emphysema. Comparable results were obtained when analyzing lungs imaged with µCT, sugges-ting that our pipeline is applicable to different kinds of imaging modalities. (4) Conclusions: We conclude that the airspace diameter map is well suited for a detailed analysis of unevenly distri-buted structural alterations in chronic muco-obstructive lung diseases such as cystic fibrosis and COPD.

A tomographic microscopy-compatible Langendorff system for the dynamic structural characterization of the cardiac cycle.

Introduction: Cardiac architecture has been extensively investigated ex vivo using a broad spectrum of imaging techniques. Nevertheless, the heart is a dynamic system and the structural mechanisms governing the cardiac cycle can only be unveiled when investigating it as such. Methods: This work presents the customization of an isolated, perfused heart system compatible with synchrotron-based X-ray phase contrast imaging (X-PCI). Results: Thanks to the capabilities of the developed setup, it was possible to visualize a beating isolated, perfused rat heart for the very first time in 4D at an unprecedented 2.75 µm pixel size (10.6 µm spatial resolution), and 1 ms temporal resolution. Discussion: The customized setup allows high-spatial resolution studies of heart architecture along the cardiac cycle and has thus the potential to serve as a tool for the characterization of the structural dynamics of the heart, including the effects of drugs and other substances able to modify the cardiac cycle.

Redescription of the Planthopper Bursinia genei (Dufour, 1849), with a New Record from Greece (Hemiptera: Fulgoromorpha: Dictyopharidae).

Bursinia genei (Dufour, 1849) is the most widespread species of the planthopper subfamily Orgeriinae (Hemiptera: Fulgoromorpha: Dictyopharidae) in Europe, found from the Iberian Peninsula to the Western Balkans. However, its diagnostically important genitalia and biology have been insufficiently described. We employ state-of-the-art synchrotron X-ray microtomography and photomicrography to re-describe B. genei, and to study the morphology of both its sexes in unprecedented detail. By examining specimens from across the distribution of B. genei, we find that they probably belong to a single, broadly distributed morphospecies. Our morphological examination allowed us to make inferences on its jumping mechanism and capacity for vibrational communication. We also record B. genei for the first time from Greece, further extending the range of this elusive species. Detailed information on the habitat of B. genei is also provided.

Wicking through complex interfaces at interlacing yarns.

HYPOTHESIS: Wicking flow in the wale direction of knit fabrics is slowed by capillary pressure minima during the transition at yarn contacts. The characteristic pore structure of yarns leads to an unfavorable free energy evolution and is the cause of these minima. EXPERIMENTS: Time-resolved synchrotron tomographic microscopy is employed to study the evolution of water configuration during wicking flow in interlacing yarns. Dynamic pore network modeling is used based on the obtained image data and distributions of delay times for pore intrusion. Good agreement is observed by comparison to the experimental data. FINDINGS: Yarn-to-yarn transition is found to coincide with slow water advance in a thin interface zone at the yarn contact. The pore spaces of the two yarns merge within this interface zone and provide a transition path. A deep capillary pressure minimum occurs while water passes through the center of the interface zone, effectively delaying the wicking flow. A pore network model considering pore intrusion delay times is expanded to include inter-yarn wicking and reproduce the observed wicking dynamics.

Mineral chemistry and 3D tomography of a Chang'E 5 high-Ti basalt: implication for the lunar thermal evolution history.

In December 2020, Chang'E-5 (CE-5), China's first lunar sample return mission, successfully collected samples totaling 1731 g from the northern Oceanus Procellarum. The landing site was located in a young mare plain, a great distance from those of Apollo and Luna missions. These young mare basalts bear critical scientific significance as they could shed light on the nature of the lunar interior (composition and structure) as well as the recent volcanism on the Moon. In this article, we investigated a CE-5 basalt sample (CE5C0000YJYX065) using a combination of state-of-art techniques, including high resolution X-ray tomographic microscopy (HR-XTM), energy dispersive X-ray spectroscopy (EDS)-based scanning electron microscope (SEM), and electron probe microanalysis (EPMA) to reveal its 3D petrology and minerology. Our results show that this sample has a fine- to medium-grained subophitic texture, with sparse olivine phenocrysts setting in the groundmass of pyroxene, plagioclase, ilmenite and trace amounts of other phases. It has an extremely high ilmenite modal abundance (17.8 vol%) and contains a significant amount (0.5 vol%) of Ca-phosphate grains. The mineral chemistry is in excellent agreement with that of Apollo and Luna high-Ti basalts. The major phase pyroxenes also display strong chemical zoning with compositions following the trends observed in Apollo high-Ti basalts. Based on current data, we came to the conclusion that CE5C0000YJYX065 is a high-Ti mare basalt with a rare earth element (REE) enriched signature. This provides a rigid ground-truth for the geological context at the CE-5 landing site and clarifies the ambiguity inferred from remote sensing surveys.

The in vitro toxicity evaluation of halloysite nanotubes (HNTs) in human lung cells.

Halloysite nanotubes (HNTs) have been increasingly used in many industrial and biomedical fields. Therefore, the assessment of risk and consequences of exposure to HNTs is very important to better protect human safety. This study aims to investigate the short- (24 or 72 h) and long-term (7 days) cytotoxic effects of HNTs at doses 10-200 µg/mL on human alveolar carcinoma epithelial cells (A549) and human bronchial epithelial cells (BEAS-2B). The effect of HNTs on cell viability, apoptosis, cell proliferation, oxidative/antioxidative status and cell morphology was evaluated. Our results showed that cytotoxicity of HNTs is dependent on dose, cell model and time of exposure. During the time of exposition toxic effects were intensified. To the best of our knowledge, this is the first study to use holo-tomographic microscopy (HTM) to visualise changes in cell morphology due to exposure from HNTs. We observed cells contraction, changes in the size and shape, cell surface folding and cytoplasmic vacuolization, peripheral arrangement of cell nuclei and even increase number of nucleus, which undoubtedly confirmed cytotoxic effect of HNTs at low doses (5 µg/mL and 25 µg/mL). Our results demonstrated that HTM technique provides a new insight into the assessment of HNTs toxicity. Further studies with different cell models are recommended to assess the toxic effect of HNTs on whole human body.

Bamboo's tissue structure facilitates large bending deflections.

Bamboo is becoming increasingly popular as an engineering material and source of bio-inspiration for instance in architecture and for the manufacture of a variety of woven products. Besides the properties of bamboo products for construction purposes, the bending deformability of thin bamboo slivers is of interest, as it appears that extraordinary large deflection can be achieved. To unravel the underlying mechanisms that may contribute to the high deformability at the tissue and cell level, bending deflection tests and additionalin situexperiments were performed to record the deflection of bamboo slivers in dependence of the tissue composition and the deformations of individual cells. For the latter, a simple bending deflection setup was used employing micro-CT measurements to analyze the deformation of individual parenchyma cells (PCs), fiber bundles and vessel elements at different stages of bending deformation of the bamboo slivers. The results showed that the degree of displacement and the characteristic fracture behavior strongly depend on the volume fractions of PCs and fibres determined by the position in the bamboo culm. For slivers with a sufficiently high fibre volume content, the very high bending deformability could be facilitated by the deformation of PCs, which are squeezed between the fibre bundles during increasing bending deflection.

A Method for Spatial Quantification of Water in Microporous Layers of Polymer Electrolyte Fuel Cells by X-ray Tomographic Microscopy.

A microporous layer (MPL) is typically added to the gas diffusion layer of polymer electrolyte fuel cells (PEFCs) to promote cell performance and water management. The transport mechanism of the water through the MPL is, however, not well understood due to its small pores (20-500 nm). Here, we demonstrate that polychromatic X-ray tomographic microscopy (XTM) can be used to determine the porosity and the spatial distribution of water in nanoporous materials and can quantitatively map the liquid water saturation of MPLs. The presented technique requires no a priori knowledge of the composition of the MPL and has a field of view on the millimeter scale, which is orders of magnitude larger than conventional electron microscopy techniques for nanoscale materials. The available field of view is compatible with existing operando cells for X-ray tomography, paving the way for the analysis of water transport in MPLs during operation.