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At least 235 records · Page 13

Honeycomb vs. Foam: Evaluating a Potential Upgrade to ISS Module Shielding for Micrometeoroids and Orbital Debris

The presence of a honeycomb core in a multi-wall shielding configuration for protection against micrometeoroid and orbital debris (MMOD) particle impacts at hypervelocity is generally considered to be detrimental as the cell walls act to restrict fragment cloud expansion, creating a more concentrated load on the shield rear wall. However, mission requirements often prevent the inclusion of a dedicated MMOD shield, and as such, structural honeycomb sandwich panels are amongst the most prevalent shield types. Open cell metallic foams are a relatively new material with novel mechanical and thermal properties that have shown promising results in preliminary hypervelocity impact shielding evaluations. In this study, an ISS-representative MMOD shielding configuration has been modified to evaluate the potential performance enhancement gained through the substitution of honeycomb for open cell foam. The baseline shielding configuration consists of a double mesh outer layer, two honeycomb sandwich panels, and an aluminum rear wall. In the modified configuration the two honeycomb cores are replaced by open-cell foam. To compensate for the heavier core material, facesheets have been removed from the second sandwich panel in the modified configuration. A total of 19 tests on the double layer honeycomb and double layer foam configurations are reported. For comparable mechanical and thermal performance, the foam modifications were shown to provide a 15% improvement in critical projectile diameter at low velocities (i.e. 3 km/s) and a 3% increase at high velocities (i.e. 7 km/s) for normal impact. With increasing obliquity, the performance enhancement was predicted to increase, up to a 29% improvement at 60 (low velocity). Ballistic limit equations have been developed for the new configuration, and consider the mass of each individual shield component in order to maintain validity in the event of minor configuration modifications. Previously identified weaknesses of open cell foams for hypervelocity impact shielding such as large projectile diameters, low velocities, and high degrees of impact obliquity have all been investigated, and found to be negligible for the double-layer configuration.

Ryan, Shannon↗

Morphological changes in woody stem of Prunus jamasakura under simulated microgravity

When the four-week-old woody stem of Prunus jamasakura was grown under simulated microgravity condition on a three-dimensional clinostat, it bent at growth, and width of its secondary xylem decreased due to the reduction of fiber cell numbers and a smaller microfibril angle in the secondary cell wall, as reported in our previous paper. Gravity induces the development of the secondary xylem that supports the stem upward against the action of gravity. In this study, morphological changes of the tissues and cells were microscopically observed. Disorder was found in the concentric structure of tissues that organize the stem. The radial arrangement of the cells was also disturbed in the secondary xylem, and in the secondary phloem secondary cell walls of the bast fiber cells were undeveloped. These findings suggest that differentiation and development of the secondary xylem and the bast fiber cells are strongly controlled by terrestrial gravity. These tissue and cells functions to support the stem under the action of gravity. Furthermore, clinorotation induced disorder in the straight joint of vessel elements and the lattice-like structure of radial parenchyma cells, which is responsible for water transportation and storage, respectively. Gravity is an essential factor for keeping the division and differentiation normal in woody stem.

Prunus/cytology/physiology/ultrastructure↗

Warming and disturbance alter soil microbiome diversity and function in a northern forest ecotone

ABSTRACT The response to global change by soil microbes is set to affect important ecosystem processes. These impacts could be most immediate in transitional zones, such as the temperate-boreal forest ecotone, yet previous work in these forests has primarily focused on specific subsets of microbial taxa. Here, we examined how bacterial and fungal communities respond to simulated above- and below-ground warming under realistic field conditions in closed and open canopy treatments in Minnesota, USA. Our results show that warming and canopy disturbance shifted bacterial and fungal community structure as dominant bacterial and fungal groups differed in the direction and intensity of their responses. Ectomycorrhizal and saprotrophic fungal communities with greater connectivity (higher prevalence of strongly interconnected taxa based on pairwise co-occurrence relationships) were more resistant to compositional change. Warming effects on soil enzymes involved in the hydrolytic and oxidative liberation of carbon from plant cell walls and nutrients from organic matter were most strongly linked to fungal community responses, although community structure–function relationships differed between fungal guilds. Collectively, these findings indicate that warming and disturbance will influence the composition and function of microbial communities in the temperate-boreal ecotone, and fungal responses are particularly important to understand for predicting future ecosystem functioning.

Microbiology↗

Additive Manufacture of Novel Tungsten Wicking and Heat Transfer Structures

Additive manufacturing (AM) is utilized for fabricating complex geometries from refractory metals, such as tungsten and molybdenum. In this study, plates with ultra-fine and fine lattice structures were manufactured from tungsten to demonstrate printability and the component fabrication advantages of AM. Previous work was completed on individual tungsten lattice structures; this work demonstrates the advantages of printing these structures on tubing and first-wall materials. Initial work using stainless steel demonstrated increased heat transfer properties promoted by these lattice structures. Applications for these structures include wicking structures, heat pipes, and first-wall materials in fusion devices. A major advantage of these designs is the capability to produce a monolithic component with tailored heat transfer and wicking properties. This research helps promote the ability to use liquid lithium in fusion devices since these lattice structures manufactured from tungsten could act as a high temperature first-wall or divertor material and as a wicking structure for liquid lithium. Different lattice structures with varying unit cell sizes and strut thicknesses were printed and the effects of these structures on heat transfer properties were measured. Abrasion resistance of these structures was also investigated. This presentation will discuss the results of these experiments and their implications related to heat transfer applications. This work will help inform the development and implementation of complex geometries made from tungsten for future heat pipe and fusion first-wall or divertor component designs.

Carly J. Romnes↗

Multiscale investigation of the mechanism of biomass deconstruction in the dimethyl isosorbide/water Co-solvent pretreatment system

In the context of promoting a circular bioeconomy, the development of green and efficient lignocellulosic biomass pretreatment technologies so as to realize high value-added biomass utilization is of intense interest. We demonstrated the potential of the bio-based green solvent dimethyl isosorbide (DMI) for the fractionation of Eucalyptus biomass with excellent performance. Here, to investigate the mechanisms involved in biomass fractionation, microimaging and microspectroscopic techniques were employed together with molecular dynamics (MD) simulation and COSMO-RS quantum chemical calculations to derive multiscale information. Both the microstructure and regional chemistry of the cell wall vary significantly with the volume ratio of DMI/H 2 O. The strongest effects were found at DMI/H 2 O = 9:1 and showed visible cell wall tearing cracks and cell wall deformation and collapse as well as the lowest values of cell wall thickness and circularity. From the MD simulations, lignin exhibits collapsed-like structure in pure H 2 O with low solvent accessibility surface area (SASA) and radius of gyration (R g ). In contrast, lignin in DMI/H 2 O shows extended structure with high SASA and solvent interactions dominated by van der Waals forces, with maximal contact in the 9:1 (v/v) system. Further, the COSMO-RS calculated sigma (σ-) potential suggests the intermolecular interactions in DMI and DMI/H 2 O co-solvent are weak, leading to stronger interaction with lignin and correspondingly higher lignin dissolution. The radial distribution functions and σ-potential all show that again DMI/H 2 O at 9:1 is an optimal volume ratio for high lignin dissolution. Finally, this study provides a solvent-ratio dependent mechanism for the action of polar aprotic solvents in the deconstruction of biomass.

09 BIOMASS FUELS↗

Lignin structural changes and high p -coumaroylation in incipient lignification in moso bamboo

Lignification is a crucial process for strengthening plant tissues, facilitating water transport, and providing defense against pathogens. In the Poaceae family, p-hydroxycinnamic acids are commonly incorporated into lignin, with acylation by p-coumarate (pCA) occurring during lignification. In this study, we performed DFRC and 2D HSQC-NMR analyses to investigate changes in lignin substructures and the degree of lignin pCA-acylation throughout bamboo stem development. Furthermore, immunohistochemical analysis was conducted to elucidate the spatial distribution of lignin substructures within different cell types. Our results revealed that, in young tissues, β–O–4-linked lignin units are predominantly derived from monolignol-pCA conjugates, specifically coniferyl- and sinapyl-pCA. Both lignin structure and the pattern of pCA acylation varied depending on the stage of cell wall formation and the cell type, particularly between vascular fiber cells and parenchyma cells. Based on our results, moso bamboo culms exhibit a distinctive feature during incipient lignification, in which monolignols are predominantly acylated with pCA. As a result, this feature has not been reported in other grasses, suggesting that extensive p-coumaroylation of monolignols plays an important role in the rapid elongation of bamboo culms.

Munekata, Noriaki [Kyoto University (Japan); Unive↗

Structure and functions of simple membrane-water interfaces

The structure and functions of the earliest ancestors of contemporary cells are focal points in studies of the origin of life. Probably the first cell-like structures were vesicles - closed, spheroidal structures with aqueous medium trapped inside. The membranous walls of vesicles were most likely bilayers composed of simple amphiphilic material available on early earth. The membrane studied was composed of glycerol 1-monooleate (GMO). Glycerol forms the polar head group and the oily tail contains 18 carbon atoms. All head groups have been found to be located in two narrow regions at the interfaces with water. The membrane interior, formed by the hydrophobic tails, is quite fluid with chain disorder increasing towards the center of the bilayer. These results are in agreement with x-ray and neutron scattering data from related bilayers. The width of the membrane is not constant, but fluctuates in time and space. Occasional thinning defects in the membrane, observed during the course of the simulations, may have a significant influence on rates of passive transport of small molecules across membranes. It has been found that water penetrates the head group region but not the oily interior of the membrane. Water molecules near the interface are oriented by dipoles of the head groups. The resulting electrostatic potential across the interface, determined in our simulations, has been found to be markedly larger than across the water-oil interface. This quantity has been implicated as the source of selectivity, with respect to the sign of the charge, as an ion approaches the interface and during transport of hydrophobic ions across membranes.

Pohorille, A.↗

Mesoscale Modeling of Dislocation Cell Structure Evolution and Radiation-Induced Segregation in Additively Manufactured Austenitic Stainless Steel

Structural alloys under irradiation develop radiation-induced segregation (RIS) at point defect sinks, which lead to undesired changes in the alloy's properties. Additively manufactured austenitic stainless steels are expected to yield a distinct response to irradiation damage owing to their unique as-printed dislocation cell structure. In this talk, we present the development of a mesoscale model for dislocation cell structure evolution coupled with RIS using the MOOSE framework. The evolution of network dislocations within spatially distinct cells and cell walls are modeled using climb-mediated edge annihilation and generation processes. Starting with initial microstructures of as-printed non-equilibrium segregations at dislocation cell walls, our 2D and 3D simulations explore the effects of irradiation temperature, dose rate, dislocation sink bias, and specimen thickness on microstructure evolution. The results will be compared against experimental characterizations of in-situ and ex-situ ion-irradiated samples, and the implications of evolving cell structure on irradiation damage response will be discussed.

additive manufacturing↗

Structure-function studies reveal ComEA contains an oligomerization domain essential for transformation in gram-positive bacteria

An essential step in bacterial transformation is the uptake of DNA into the periplasm, across the thick peptidoglycan cell wall of Gram-positive bacteria, or the outer membrane and thin peptidoglycan layer of Gram-negative bacteria. ComEA, a DNA-binding protein widely conserved in transformable bacteria, is required for this uptake step. Here we determine X-ray crystal structures of ComEA from two Gram-positive species, Bacillus subtilis and Geobacillus stearothermophilus, identifying a domain that is absent in Gram-negative bacteria. X-ray crystallographic, genetic, and analytical ultracentrifugation (AUC) analyses reveal that this domain drives ComEA oligomerization, which we show is required for transformation. We use multi-wavelength AUC (MW-AUC) to characterize the interaction between DNA and the ComEA DNA-binding domain. Finally, we present a model for the interaction of the ComEA DNA-binding domain with DNA, suggesting that ComEA oligomerization may provide a pulling force that drives DNA uptake across the thick cell walls of Gram-positive bacteria.

59 BASIC BIOLOGICAL SCIENCES↗

Pseudomonas aeruginosa C-Terminal Processing Protease CtpA Assembles into a Hexameric Structure That Requires Activation by a Spiral-Shaped Lipoprotein-Binding Partner

Pseudomonas aeruginosa CtpA is a carboxyl-terminal processing protease that partners with the outer membrane lipoprotein LbcA to degrade at least five cell wall-associated proteins, four of which are cell wall hydrolases. This activity plays an important role in supporting P. aeruginosa virulence in a mouse model of acute pneumonia. However, almost nothing is known about the molecular mechanisms underlying CtpA and LbcA function. Here, we used structural analysis to show that CtpA alone assembles into an inactive hexamer comprising a trimer of dimers, which limits its substrate access and prevents nonspecific degradation. The adaptor protein LbcA is a right-handed open spiral with 11 tetratricopeptide repeats, which might wrap around a substrate to deliver it to CtpA for degradation. By structure-guided mutagenesis and functional assays, we also showed that the interfaces of the CtpA trimer of dimers and an N-terminal helix of LbcA are important for LbcA-mediated substrate degradation by CtpA both in vitro and in vivo. This work improves our understanding of the molecular mechanism of the LbcA-CtpA proteolytic system and reveals some striking differences from the arrangements found in some other bacterial CTPs.

structural biology↗

Transcriptome Mining Provides Insights into Cell Wall Metabolism and Fiber Lignification in Agave tequilana Weber

Resilience of growing in arid and semiarid regions and a high capacity of accumulating sugar-rich biomass with low lignin percentages have placed Agave species as an emerging bioenergy crop. Although transcriptome sequencing of fiber-producing agave species has been explored, molecular bases that control wall cell biogenesis and metabolism in agave species are still poorly understood. Here, through RNAseq data mining, we reconstructed the cellulose biosynthesis pathway and the phenylpropanoid route producing lignin monomers in A. tequilana, and evaluated their expression patterns in silico and experimentally. Most of the orthologs retrieved showed differential expression levels when they were analyzed in different tissues with contrasting cellulose and lignin accumulation. Phylogenetic and structural motif analyses of putative CESA and CAD proteins allowed to identify those potentially involved with secondary cell wall formation. RT-qPCR assays revealed enhanced expression levels of AtqCAD5 and AtqCESA7 in parenchyma cells associated with extraxylary fibers, suggesting a mechanism of formation of sclerenchyma fibers in Agave similar to that reported for xylem cells in model eudicots. Overall, our results provide a framework for understanding molecular bases underlying cell wall biogenesis in Agave species studying mechanisms involving in leaf fiber development in monocots.

59 BASIC BIOLOGICAL SCIENCES↗

Evidence for Lignin–Carbohydrate Complexes from Studies of Transgenic Switchgrass and a Model Lignin–Pectin Composite

Lignin–carbohydrate complexes (LCCs) form through interactions of lignin with plant cell wall polysaccharides and are thought to be a significant source of biomass recalcitrance. In this work, we investigated LCCs formed between lignin and pectin homogalacturonan (HG). The structural changes in HG deficient transgenic switchgrass (GAUT4-knockdown, GAUT4-KD) after hot water pretreatment were compared to wild-type plants using small-angle neutron scattering (SANS), which showed that there were ~2.2-fold more lignin aggregates in GAUT4-KD biomass compared to the wild type. This demonstrated that decreased pectin resulted in more lignin redistribution and suggested that interactions between lignin and HG restrict lignin mobility in plant cell walls. To better understand the types of interactions between lignin and pectin, a model composite was prepared by polymerizing either protiated or partially deuterated coniferyl alcohol to form a dehydrogenation polymer (DHP) in the presence of HG. Small-angle X-ray scattering (SAXS) showed that the DHP and HG form a highly interconnected network structure that is not observed in a physical mixture of the individual polymers. Contrast matching SANS revealed the structure of DHP and HG in the composite and showed that the HG forms a swollen interconnected polymer network (power-law exponent, P = 1.5) interspersed with DHP particles (radius of gyration, Rg, 264 Å) that are composed of solvent-accessible DHP polymers (P = 2.3). Fourier transform infrared spectroscopy showed a unique ester absorption band in the DHP/HG composites. Solid-state nuclear magnetic resonance (NMR) analysis also supports interactions between DHP and HG. Overall, this study provides new insights into the relationship between primary and secondary cell wall polymers during cell wall synthesis and how LCCs formed between pectin and lignin could represent a previously unrecognized source of biomass recalcitrance. This knowledge may help develop new approaches to modulate cell wall properties to improve biofuel and bioproduct production.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

New genes and new biological roles for expansins

Expansins are extracellular proteins that loosen plant cell walls in novel ways. They are thought to function in cell enlargement, pollen tube invasion of the stigma (in grasses), wall disassembly during fruit ripening, abscission and other cell separation events. Expansins are encoded by two multigene families and each gene is often expressed in highly specific locations and cell types. Structural analysis indicates that one expansin region resembles the catalytic domain of family-45 endoglucanases but glucanase activity has not been detected. The genome projects have revealed numerous expansin-related sequences but their putative wall-loosening functions remain to be assessed.

NASA Program Fundamental Space Biology↗

Mathematical modeling of the flow field and particle motion in a rotating bioreactor at unit gravity and microgravity

The biotechnology group at NASA Johnson Space Center is developing systems for culturing mammalian cells that stimulate some aspect of microgravity and provide a low shear environment for microgravity-based studies on suspension and anchorage dependent cells. The design of these vessels for culturing cells is based on the need to suspend cells and aggregates of cells and microcarrier beads continually in the culturing medium. The design must also provide sufficient circulation for adequate mass transfer of nutrients to the cells and minimize the total force on the cells. Forces, resulting from sources such as hydrodynamic fluid shear and collisions of cells and walls of the vessels, may damage delicate cells and degrade the formation of three dimensional structures. This study examines one particular design in both unit gravity and microgravity based on two concentric cylinders rotating in the same direction at different speeds to create a Couette flow between them. A numerical simulation for the flow field and the trajectories of particles in the vessel. The flow field for the circulation of the culturing medium is modeled by the Navier-Stokes equations. The forces on a particle are assumed to be drag from the fluid's circulation, buoyancy from the gravitational force and centrifugal force from the rotation of the vessel. The problem requires first solving the system of partial differential equations for the fluid flow by a finite difference method and then solving the system of ordinary differential equations for the trajectories by Gear's stiff method. Results of the study indicate that the trajectories in unit gravity and microgravity are very similar except for small spatial deviations on the fast time scale in unit gravity. The total force per unit cross sectional area on a particle in microgravity, however, is significantly smaller than the corresponding value in unit gravity, which is also smaller than anticipated. Hence, this study indicates that this design for a bioreactor with optimal rates of rotation can provide a good environment for culturing cells in microgravity with adequate circulation and minimal force on the cells.

Boyd, Ernest J.↗

Tailoring renewable materials via plant biotechnology

Plants inherently display a rich diversity in cell wall chemistry, as they synthesize an array of polysaccharides along with lignin, a polyphenolic that can vary dramatically in subunit composition and interunit linkage complexity. These same cell wall chemical constituents play essential roles in our society, having been isolated by a variety of evolving industrial processes and employed in the production of an array of commodity products to which humans are reliant. However, these polymers are inherently synthesized and intricately packaged into complex structures that facilitate plant survival and adaptation to local biogeoclimatic regions and stresses, not for ease of deconstruction and commercial product development. Herein, we describe evolving techniques and strategies for altering the metabolic pathways related to plant cell wall biosynthesis, and highlight the resulting impact on chemistry, architecture, and polymer interactions. Furthermore, this review illustrates how these unique targeted cell wall modifications could significantly extend the number, diversity, and value of products generated in existing and emerging biorefineries. These modifications can further target the ability for processing of engineered wood into advanced high performance materials. In doing so, we attempt to illuminate the complex connection on how polymer chemistry and structure can be tailored to advance renewable material applications, using all the chemical constituents of plant-derived biopolymers, including pectins, hemicelluloses, cellulose, and lignins.

09 BIOMASS FUELS↗

Sensor Domain of Histidine Kinase VxrA of Vibrio cholerae : Hairpin-Swapped Dimer and Its Conformational Change

VxrA and VxrB are cognate histidine kinase (HK)-response regulator (RR) pairs of a two-component signaling system (TCS) found in Vibrio cholerae, a bacterial pathogen that causes cholera. The VxrAB TCS positively regulates virulence, the type VI secretion system, biofilm formation, and cell wall homeostasis in V. cholerae, providing protection from environmental stresses and contributing to the transmission and virulence of the pathogen. The VxrA HK has a unique periplasmic sensor domain (SD) and, remarkably, lacks a cytoplasmic linker domain between the second transmembrane helix and the dimerization and histidine phosphotransfer (DHp) domain, indicating that this system may utilize a potentially unique signal sensing and transmission TCS mechanism. Here, we have determined several crystal structures of VxrA-SD and its mutants. These structures reveal a novel structural fold forming an unusual β hairpin-swapped dimer. A conformational change caused by relative rotation of the two monomers in a VxrA-SD dimer could potentially change the association of transmembrane helices and, subsequently, the pairing of cytoplasmic DHp domains. Based on the structural observation, we propose a putative scissor-like closing regulation mechanism for the VxrA HK. IMPORTANCE V. cholerae has a dynamic life cycle, which requires rapid adaptation to changing external conditions. Two-component signal transduction (TCS) systems allow V. cholerae to sense and respond to these environmental changes. The VxrAB TCS positively regulates a number of important V. cholerae phenotypes, including virulence, the type six secretion system, biofilm formation, and cell wall homeostasis. Here, we provide the crystal structures of the VxrA sensor histidine kinase sensing domain and propose a mechanism for signal transduction. The cognate signal for VxrAB remains unknown; however, in this work we couple our structural analysis with functional assessments of key residues to further our understanding of this important TCS.

59 BASIC BIOLOGICAL SCIENCES↗

Molecular regulation of plant cell wall extensibility

Gravity responses in plants often involve spatial and temporal changes in cell growth, which is regulated primarily by controlling the ability of the cell wall to extend. The wall is thought to be a cellulose-hemicellulose network embedded in a hydrated matrix of complex polysaccharides and a small amount of structural protein. The wall extends by a form of polymer creep, which is mediated by expansins, a novel group of wall-loosening proteins. Expansins were discovered during a molecular dissection of the "acid growth" behavior of cell walls. Expansin alters the rheology of plant walls in profound ways, yet its molecular mechanism of action is still uncertain. It lacks detectable hydrolytic activity against the major components of the wall, but it is able to disrupt noncovalent adhesion between wall polysaccharides. The discovery of a second family of expansins (beta-expansins) sheds light on the biological role of a major group of pollen allergens and implies that expansins have evolved for diverse developmental functions. Finally, the contribution of other processes to wall extensibility is briefly summarized.

Review↗

Architecture of a catalytically active homotrimeric plant cellulose synthase complex

Cellulose is an essential plant cell wall component and represents the most abundant biopolymer on Earth. Supramolecular plant cellulose synthase complexes organize multiple linear glucose polymers into microfibrils as load-bearing wall components. We determined the structure of a poplar cellulose synthase CesA homotrimer that suggests a molecular basis for cellulose microfibril formation. This complex, stabilized by cytosolic plant-conserved regions and helical exchange within the transmembrane segments, forms three channels occupied by nascent cellulose polymers. Secretion steers the polymers toward a common exit point, which could facilitate protofibril formation. CesA’s N-terminal domains assemble into a cytosolic stalk that interacts with a microtubule-tethering protein and may thus be involved in CesA localization. Here, our data suggest how cellulose synthase complexes assemble and provide the molecular basis for plant cell wall engineering.

59 BASIC BIOLOGICAL SCIENCES↗