Cardiomyocyte orientation recovery at micrometer scale reveals long-axis fiber continuum in heart walls.

Coordinated cardiomyocyte contraction drives the mammalian heart to beat and circulate blood. No consensus model of cardiomyocyte geometrical arrangement exists, due to the limited spatial resolution of whole heart imaging methods and the piecemeal nature of studies based on histological sections. By combining microscopy and computer vision, we produced the first-ever three-dimensional cardiomyocyte orientation reconstruction across mouse ventricular walls at the micrometer scale, representing a gain of three orders of magnitude in spatial resolution. We recovered a cardiomyocyte arrangement aligned to the long-axis direction of the outer ventricular walls. This cellular network lies in a thin shell and forms a continuum with longitudinally arranged cardiomyocytes in the inner walls, with a complex geometry at the apex. Our reconstruction methods can be applied at fine spatial scales to further understanding of heart wall electrical function and mechanics, and set the stage for the study of micron-scale fiber remodeling in heart disease.

Cell wall dynamics: novel tools and research questions.

Years ago, a classic textbook would define plant cell walls based on passive features. For instance, a sort of plant exoskeleton of invariable polysaccharide composition, and probably painted in green. However, currently, this view has been expanded to consider plant cell walls as active, heterogeneous, and dynamic structures with a high degree of complexity. However, what do we mean when we refer to a cell wall as a dynamic structure? How can we investigate the different implications of this dynamism? While the first question has been the subject of several recent publications, defining the ideal strategies and tools needed to address the second question has proven to be challenging due to the myriad of techniques available. In this review, we will describe the capacities of several methodologies to study cell wall composition, structure, and other aspects developed or optimized in recent years. Keeping in mind cell wall dynamism and plasticity, the advantages of performing long-term non-invasive live-imaging methods will be emphasized. We specifically focus on techniques developed for Arabidopsis thaliana primary cell walls, but the techniques could be applied to both secondary cell walls and other plant species. We believe this toolset will help researchers in expanding knowledge of these dynamic/evolving structures.

Radial microfibril arrangements in wood cell walls.

MAIN CONCLUSION: TEM and AFM imaging reveal radial orientations and whorl-like arrangements of cellulose microfibrils near the S1/S2 interface. These are explained by wrinkling during lamellar cell growth. In the most widely accepted model of the ultrastructure of wood cell walls, the cellulose microfibrils are arranged in helical patterns on concentric layers. However, this model is contradicted by a number of transmission electron microscopy (TEM) studies which reveal a radial component to the microfibril orientations in the cell wall. The idea of a radial component of the microfibril directions is not widely accepted, since it cannot easily be explained within the current understanding of lamellar cell growth. To help clarify the microfibril arrangements in wood cell walls, we have investigated various wood cell wall sections using both transmission electron microscopy and atomic force microscopy, and using various imaging and specimen preparation methods. Our investigations confirm that the microfibrils have a radial component near the interface between the S1 and S2 cell wall layers, and also reveal a whorl-like microfibril arrangement at the S1/S2 interface. These whorl-like structures are consistent with cell wall wrinkling during growth, allowing the radial microfibril component to be reconciled with the established models for lamellar cell growth.

Brussowvirus SW13 Requires a Cell Surface-Associated Polysaccharide To Recognize Its Streptococcus thermophilus Host.

Four bacteriophage-insensitive mutants (BIMs) of the dairy starter bacterium Streptococcus thermophilus UCCSt50 were isolated following challenge with Brussowvirus SW13. The BIMs displayed an altered sedimentation phenotype. Whole-genome sequencing and comparative genomic analysis of the BIMs uncovered mutations within a family 2 glycosyltransferase-encoding gene (orf06955UCCSt50) located within the variable region of the cell wall-associated rhamnose-glucose polymer (Rgp) biosynthesis locus (designated the rgp gene cluster here). Complementation of a representative BIM, S. thermophilus B1, with native orf06955UCCSt50 restored phage sensitivity comparable to that of the parent strain. Detailed bioinformatic analysis of the gene product of orf06955UCCSt50 identified it as a functional homolog of the Lactococcus lactis polysaccharide pellicle (PSP) initiator WpsA. Biochemical analysis of cell wall fractions of strains UCCSt50 and B1 determined that mutations within orf06955UCCSt50 result in the loss of the side chain decoration from the Rgp backbone structure. Furthermore, it was demonstrated that the intact Rgp structure incorporating the side chain structure is essential for phage binding through fluorescence labeling studies. Overall, this study confirms that the rgp gene cluster of S. thermophilus encodes the biosynthetic machinery for a cell surface-associated polysaccharide that is essential for binding and subsequent infection by Brussowviruses, thus enhancing our understanding of S. thermophilus phage-host dynamics. IMPORTANCE Streptococcus thermophilus is an important starter culture bacterium in global dairy fermentation processes, where it is used for the production of various cheeses and yogurt. Bacteriophage predation of the species can result in substandard product quality and, in rare cases, complete fermentation collapse. To mitigate these risks, it is necessary to understand the phage-host interaction process, which commences with the recognition of, and adsorption to, specific host-encoded cell surface receptors by bacteriophage(s). As new groups of S. thermophilus phages are being discovered, the importance of underpinning the genomic elements that specify the surface receptor(s) is apparent. Our research identifies a single gene that is critical for the biosynthesis of a saccharidic moiety required for phage adsorption to its S. thermophilus host. The acquired knowledge provides novel insights into phage-host interactions for this economically important starter species.

How to turn the corner: Discrimination of path shapes in rats.

The internal representation of a path shape is an element that constructs an internal representation of an entire route or environment. In the present study, we examined the ability of rats to discriminate path shapes. The rats learned to discriminate between an oval-shaped runway and a square-shaped one and to respond to one of two response boxes on the two sides of the runways. After the learning sessions, we tested which of the inner and outer walls the rats used as cues for discrimination using different wall shapes. The results suggest that the rats used the shape of the inner walls for the discrimination. Subsequently, the learning sessions, in which different shapes of the inner and outer walls were used, continued. There was a tendency for the rats to show better performance when the shape of the inner walls was congruent with the rule in the original learning, suggesting again that the rats used the shape of the inner wall for the discrimination. In addition, similar results were obtained when the task was conducted in the dark, suggesting that rats can discriminate path shapes using non-visual information.

Cell wall modification and lignin biosynthesis involved in disease resistance against Diaporthe citri in harvested pummelo fruit elicited by carvacrol.

BACKGROUND: Phomopsis stem-end rot caused by Diaporthe citri, causes significant commercial postharvest losses of pummelo fruit during storage. Carvacrol (CVR) is a known generally recognized as safe and has the ability to prolong the preservation of harvested fruits. In the present study, the inhibitory effects of CVR treatment at the appropriate concentration on Phomopsis stem-end rot development of harvested pummelo fruit inoculated with D. citri were evaluated by the amounts of cell wall components, the activities and gene expressions of related enzymes involved in cell wall modification and lignin biosynthesis. RESULTS: Results indicated that CVR completely inhibited D. citri growth in vitro at 200 mg L-1 and significantly controlled Phomopsis stem-end rot development in harvested pummelo. The CVR treatment delayed peel softening and browning, and retarded electrolyte leakage, superoxide radical (O2 •- ) production, and malondialdehyde content. The CVR-treated fruit maintained higher amounts of cell wall material, protopectin, hemicelluloses, and cellulose, but exhibited lower water-soluble pectin amount. Moreover, in D. citri-inoculated fruit, CVR treatment suppressed the activities and gene expressions of cell wall disassembling-enzymes, including pectin methylesterase, polygalacturonase, cellulase, and ß-galactosidase, while the development of cell wall degradation was reduced. Meanwhile, the CVR treatment enhanced the lignin biosynthesis by increasing the activities and up-regulating the gene expressions of phenylalanine ammonialyase, cinnamic alcohol dehydrogenase, and peroxidase accompanied with elevated level of lignin in pummelo fruit. CONCLUSION: The disease resistance to D. citri in pummelo fruit elicited by CVR treatment is related to delaying cell wall degradation and enhancing lignin biosynthesis. © 2021 Society of Chemical Industry.

Impact of the Environment upon the Candida albicans Cell Wall and Resultant Effects upon Immune Surveillance.

The fungal cell wall is an essential organelle that maintains cellular morphology and protects the fungus from environmental insults. For fungal pathogens such as Candida albicans, it provides a degree of protection against attack by host immune defences. However, the cell wall also presents key epitopes that trigger host immunity and attractive targets for antifungal drugs. Rather than being a rigid shield, it has become clear that the fungal cell wall is an elastic organelle that permits rapid changes in cell volume and the transit of large liposomal particles such as extracellular vesicles. The fungal cell wall is also flexible in that it adapts to local environmental inputs, thereby enhancing the fitness of the fungus in these microenvironments. Recent evidence indicates that this cell wall adaptation affects host-fungus interactions by altering the exposure of major cell wall epitopes that are recognised by innate immune cells. Therefore, we discuss the impact of environmental adaptation upon fungal cell wall structure, and how this affects immune recognition, focussing on C. albicans and drawing parallels with other fungal pathogens.

Hevea brasiliensis coniferaldehyde-5-hydroxylase (HbCAld5H) regulates xylogenesis, structure and lignin chemistry of xylem cell wall in Nicotiana tabacum.

KEY MESSAGE: The HbCAld5H1 gene cloned from Hevea brasiliensis regulates the cambial activity, xylem differentiation, syringyl-guaiacyl ratio, secondary wall structure, lignification pattern and xylan distribution in xylem fibres of transgenic tobacco plants. Molecular characterization of lignin biosynthesis gene coniferaldehyde-5-hydroxylase (CAld5H) from Hevea brasiliensis and its functional validation was performed. Both sense and antisense constructs of HbCAld5H1 gene were introduced into tobacco through Agrobacterium-mediated genetic transformation for over expression and down-regulation of this key enzyme to understand its role affecting structural and cell wall chemistry. The anatomical studies of transgenic tobacco plants revealed the increase of cambial activity leading to xylogenesis in sense lines and considerable reduction in antisense lines. The ultra-structural studies showed that the thickness of secondary wall (S2 layer) of fibre had been decreased with non-homogenous lignin distribution in antisense lines, while sense lines showed an increase in S2 layer thickness. Maule color reaction revealed that syringyl lignin distribution in the xylem elements was increased in sense and decreased in antisense lines. The immunoelectron microscopy revealed a reduction in LM 10 and LM 11 labelling in the secondary wall of antisense tobacco lines. Biochemical studies showed a radical increase in syringyl lignin in sense lines without any significant change in total lignin content, while S/G ratio decreased considerably in antisense lines. Our results suggest that CAld5H gene plays an important role in xylogenesis stages such as cambial cell division, secondary wall thickness, xylan and syringyl lignin distribution in tobacco. Therefore, CAld5H gene could be considered as a promising target for lignin modification essential for timber quality improvement in rubber.

Even Visually Intact Cell Walls in Waterlogged Archaeological Wood Are Chemically Deteriorated and Mechanically Fragile: A Case of a 170 Year-Old Shipwreck.

Structural and chemical deterioration and its impact on cell wall mechanics were investigated for visually intact cell walls (VICWs) in waterlogged archaeological wood (WAW). Cell wall mechanical properties were examined by nanoindentation without prior embedding. WAW showed more than 25% decrease of both hardness and elastic modulus. Changes of cell wall composition, cellulose crystallite structure and porosity were investigated by ATR-FTIR imaging, Raman imaging, wet chemistry, 13C-solid state NMR, pyrolysis-GC/MS, wide angle X-ray scattering, and N2 nitrogen adsorption. VICWs in WAW possessed a cleavage of carboxyl in side chains of xylan, a serious loss of polysaccharides, and a partial breakage of ß-O-4 interlinks in lignin. This was accompanied by a higher amount of mesopores in cell walls. Even VICWs in WAW were severely deteriorated at the nanoscale with impact on mechanics, which has strong implications for the conservation of archaeological shipwrecks.

The Effect of Thermomechanical Pretreatment on the Structure and Properties of Lignin-Rich Plant Biomass.

The cooperative thermomechanical properties of plant-derived polymers have been studied insufficiently, although this feedstock has a very high potential. In the present paper, we analyzed the changes in the structure and physicochemical properties of lignin-rich biomass induced by thermomechanical pretreatment. Low-temperature treatment allows one to retain the original supramolecular structure of the cell walls, while an appreciably high disintegration degree is reached. This increases the reactivity of the material in the subsequent heterogeneous reactions. Mechanical pretreatment at medium temperatures (10 °C), when almost all cell wall polymers except for low-molecular-weight lignin are in the glassy state, enhances the mobility of cell wall polymers and causes sufficient cellulose disordering, while the specific surface area is not significantly increased. High-temperature pretreatment of reed biomass is accompanied by pore formation and lignin release from the cell wall structure, which opens up new prospects for using this biomass as a matrix to produce core-shell-structured sorbents of heavy metals. The energy consumed by mechanochemical equipment for the activation of reed biomass was determined.

Peculiar features of aortic wall structure among children with coarctation.

OBJECTIVE: The aim of the study to evaluate the peculiarities of the aortic wall structure at the place of coarctation. PATIENTS AND METHODS: Materials and methods: Studying of the aortic sections removed during operative correction at the place of constriction. 10 children at the age between 1 to 6 months were undergone the operation. Intraoperative aortic biopsy specimens were observed in 10% neutral formalin. Histologic sections were prepared in a conventional way followed by staining them with hematoxylin-eosin. RESULTS: Results: Histological examination in the areas of constriction revealed that the endothelium in all the preparations had poor expressiveness. The most significant changes were recorded in the middle layer of the aorta in the form of reduced development of elastic fibers, their fragmentation and chaotic arrangement. Angiomatosis with the formation of thin-layer small vessels by capillary type was found out. In all the preparations, areas of emptying of cells and fibers of the middle cover with the formation of cystic structures were revealed. CONCLUSION: Conclusions: The histological examination has revealed changes in the structure of the aorta wall, which may indicate the systemic nature of the lesion and make it possible to consider coarctation of the aorta to be a manifestation of systemic vasculopathy. The above-mentioned facts determine the need for a more detailed examination of children with the specified pathology at different stages of observation.

Specificity of morphofunctional structure of the human and animal biliary system (analytical literature review).

OBJECTIVE: The aim of the paper was the study of the structure of the basic parts of the gallbladder wall of humans and animals of different classes in a comparatively anatomical aspect using the bibliographic analysis of literature sources. PATIENTS AND METHODS: Materials and methods: of the study was the contemporary publications on the anatomical and histological structure of the human and animal gallbladder wall mucosa, studied using the analysis, synthesis and summarizing of the obtained information. CONCLUSION: Conclusions: Basically, the structure of the gallbladder wall mucosa in humans and animals of different classes has much in common; however there are also significant differences depending on the natural type of nutrition. Structural organization of the gallbladder wall in animals in comparison with humans has individual features and depends on the type of nutrition.

Xyloglucan in the primary cell wall: assessment by FESEM, selective enzyme digestions and nanogold affinity tags.

Xyloglucan has been hypothesized to bind extensively to cellulose microfibril surfaces and to tether microfibrils into a load-bearing network, thereby playing a central role in wall mechanics and growth, but this view is challenged by newer results. Here we combined high-resolution imaging by field emission scanning electron microscopy (FESEM) with nanogold affinity tags and selective endoglucanase treatments to assess the spatial location and conformation of xyloglucan in onion cell walls. FESEM imaging of xyloglucanase-digested cell walls revealed an altered microfibril organization but did not yield clear evidence of xyloglucan conformations. Backscattered electron detection provided excellent detection of nanogold affinity tags in the context of wall fibrillar organization. Labelling with xyloglucan-specific CBM76 conjugated with nanogold showed that xyloglucans were associated with fibril surfaces in both extended and coiled conformations, but tethered configurations were not observed. Labelling with nanogold-conjugated CBM3, which binds the hydrophobic surface of crystalline cellulose, was infrequent until the wall was predigested with xyloglucanase, whereupon microfibril labelling was extensive. When tamarind xyloglucan was allowed to bind to xyloglucan-depleted onion walls, CBM76 labelling gave positive evidence for xyloglucans in both extended and coiled conformations, yet xyloglucan chains were not directly visible by FESEM. These results indicate that an appreciable, but still small, surface of cellulose microfibrils in the onion wall is tightly bound with extended xyloglucan chains and that some of the xyloglucan has a coiled conformation.

Cell wall dynamics and gene expression on soybean embryonic axes during germination.

MAIN CONCLUSION: Identification of the structural changes and cell wall-related genes likely involved in cell wall extension, cellular water balance and cell wall biosynthesis on embryonic axes during germination of soybean seeds. Cell wall is a highly organized and dynamic structure that provides mechanical support for the cell. During seed germination, the cell wall is critical for cell growth and seedling establishment. Although seed germination has been widely studied in several species, key aspects regarding the regulation of cell wall dynamics in germinating embryonic axes remain obscure. Here, we characterize the gene expression patterns of cell wall pathways and investigate their impact on the cell wall dynamics of embryonic axes of germinating soybean seeds. We found 2143 genes involved in cell wall biosynthesis and assembly in the soybean genome. Key cell wall genes were highly expressed at specific germination stages, such as expansins, UDP-Glc epimerases, GT family, cellulose synthases, peroxidases, arabinogalactans, and xyloglucans-related genes. Further, we found that embryonic axes grow through modulation of these specific cell wall genes with no increment in biomass. Cell wall structural analysis revealed a defined pattern of cell expansion and an increase in cellulose content during germination. In addition, we found a clear correlation between these structural changes and expression patterns of cell wall genes during germination. Taken together, our results provide a better understanding of the complex transcriptional regulation of cell wall genes that drive embryonic axes growth and expansion during soybean germination.

Factors influencing thermal injury to skin and abdominal wall structures in HIFU ablation of uterine fibroids.

OBJECTIVE: To investigate the factors which may cause thermal injury of abdominal wall structures in ultrasound-guided high-intensity focussed ultrasound (USgHIFU) ablation of uterine fibroids. METHOD: A total of 892 patients with uterine fibroids diagnosed on contrast-enhanced magnetic resonance imaging (MRI) scans received HIFU ablation and follow-up MRI scanning. After therapy, thermal injury to the skin was assessed via measurement of skin redness, blisters, subcutaneous nodules and to the abdominal wall structures via measurement of signal intensity on T2-weighted MRI images. A total of 151 patients were assigned to the injury group, 741 patients were assigned to the non-injury group. The relationship between patient and treatment parameters and injury were analysed using univariate and multiple logistic regression analyses. RESULTS: Univariate logistic regression revealed that sonication time, sonication time per hour, total energy deposited, distance from uterine fibroid ventral side to skin, volume of uterine fibroids, abdominal wall scar, abdominal wall thickness and body mass index (BMI) all affected whether thermal injury occurred (p < 0.05). Subsequently, multiple logistic regression analysis revealed that total energy (p = 0.000, OR = 2.228, 95% CI 1.831-2.712), abdominal wall scar (p = 0.019, OR = 1.639, 95% CI 1.085-2.477) and abdominal wall thickness (p = 0.000, OR = 1.562, 95% CI 1.313-1.857) were significantly correlated with thermal injury. CONCLUSION: Multiple logistic regression analysis revealed that abdominal wall thickness, total energy and abdominal wall scar were the most significant influencing factors that influenced minimal thermal injury of abdominal wall structures in USgHIFU ablation of uterine fibroids.

Boron increases root elongation by reducing aluminum induced disorganized distribution of HG epitopes and alterations in subcellular cell wall structure of trifoliate orange roots.

Aluminum toxicity limits the plant growth by inducing inhibition of root elongation. Although several mechanisms have been proposed regarding the phytotoxic effects of aluminum on inhibition of root elongation; the primary causes of aluminum induced root inhibition and its mitigation by boron (B) are still elusive. The present study was carried out to explore the mechanisms of B induced mitigation of aluminum toxicity and to investigate the changes in well wall structure under aluminum toxicity coupled with the techniques of confocal laser microscope, lumogallion and transmission electron microscope. The results revealed that aluminum toxicity severely hampered the root elongation and plant biomass. Moreover, alteration in subcellular structure were observed under aluminum toxicity, however, such negative effects were further exacerbated with B deficiency. Aluminum toxicity indicated disorganized distribution of HG (homogalacturonan) epitopes with higher accumulation of apoplastic aluminum. Nevertheless, B supply improved root elongation, and reduced the aluminum uptake. Taken together, it is concluded that B application can reduce aluminum toxicity and improve root elongation by decreasing Al3+ accumulation to cell wall, alteration in the cell wall structure and reducing the distribution of HG epitopes in the roots of trifoliate (Poncirus trifoliate) orange.

Gradient of structural traits drives hygroscopic movements of scarious bracts surrounding Helichrysum bracteatum capitulum.

Background and Aims: The capitulum of Helichrysum bracteatum is surrounded by scarious involucral bracts that perform hygroscopic movements leading to bract bending toward or away from the capitulum, depending on cell wall water status. The present investigation aimed at explaining the mechanism of these movements. Methods: Surface strain and bract shape changes accompanying the movements were quantified using the replica method. Dissection experiments were used to assess the contribution of different tissues in bract deformation. Cell wall structure and composition were examined with the aid of light and electron microscopy as well as confocal Raman spectroscopy. Key Results: At the bract hinge (organ actuator) longitudinal strains at opposite surfaces differ profoundly. This results in changes of hinge curvature that drive passive displacement of distal bract portions. The distal portions in turn undergo nearly uniform strain on both surfaces and also minute shape changes. The hinge is built of sclerenchyma-like abaxial tissue, parenchyma and adaxial epidermis with thickened outer walls. Cell wall composition is rather uniform but tissue fraction occupied by cell walls, cell wall thickness, compactness and cellulose microfibril orientation change gradually from abaxial to adaxial hinge surface. Dissection experiments show that the presence of part of the hinge tissues is enough for movements. Conclusions: Differential strain at the hinge is due to adaxial-abaxial gradient in structural traits of hinge tissues and cell walls. Thus, the bract hinge of H. bracteatum is a structure comprising gradually changing tissues, from highly resisting to highly active, rather than a bi-layered structure with distinct active and resistance parts, often ascribed for hygroscopically moving organs.

Effect of Thermal and Nonthermal Processing on Textural Quality of Plant Tissues.

In the current fast revolving world, the consumption of processed food is increasing drastically. The population who depend on these processed foods are also cautious about the quality and safety of what they consume. This being the case, in order to satisfy the consumer it is the responsibility of the researcher and the manufacturer to check what happens to food on processing. Plant-derived foods such as fruits and vegetables are sensitive producers which are to be handled cautiously through each steps involved in processing, starting from harvest to storage, processing to package, transportation to distribution, till it reaches the consumer. During processing, the plant materials, which are made up of complex structural components such as lignin, cellulose, pectin, etc. undergo changes which has its effect on the quality attributes of the final product. Texture is an important quality parameter of all the sensory properties. The relation between the structure of the plant tissue and the texture of the final product is reviewed in this paper comprehensively.

Seed set, pollen morphology and pollen surface composition response to heat stress in field pea.

Pea (Pisum sativum L.) is a major legume crop grown in a semi-arid climate in Western Canada, where heat stress affects pollination, seed set and yield. Seed set and pod growth characteristics, along with in vitro percentage pollen germination, pollen tube growth and pollen surface composition, were measured in two pea cultivars (CDC Golden and CDC Sage) subjected to five maximum temperature regimes ranging from 24 to 36 °C. Heat stress reduced percentage pollen germination, pollen tube length, pod length, seed number per pod, and the seed-ovule ratio. Percentage pollen germination of CDC Sage was greater than CDC Golden at 36 °C. No visible morphological differences in pollen grains or the pollen surface were observed between the heat and control-treated pea. However, pollen wall (intine) thickness increased due to heat stress. Mid-infrared attenuated total reflectance (MIR-ATR) spectra revealed that the chemical composition (lipid, proteins and carbohydrates) of each cultivar's pollen grains responded differently to heat stress. The lipid region of the pollen coat and exine of CDC Sage was more stable compared with CDC Golden at 36 °C. Secondary derivatives of ATR spectra indicated the presence of two lipid types, with different amounts present in pollen grains from each cultivar.