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51 records · Page 3

Changes in Mouse Bone Turnover in Response to Microgravity

Mechanical unloading during spaceflight is known to adversely affect mammalian physiology. Our previous studies using the Animal Enclosure Module on short duration Shuttle missions enabled us to identify a deficit in stem cell based-tissue regeneration as being a significant concern for long-duration spaceflight. Specifically, we found that mechanical unloading in microgravity resulted in inhibition of differentiation of mesenchymal and hematopoietic stem cells in the bone marrow compartment. Also, we observed overexpression of a cell cycle arrest molecule, CDKN1a/p21, in osteoprecursor cells on the bone surface, chondroprogenitors in the articular cartilage, and in myofibers attached to bone tissue. Specifically in bone tissue during both short (15-day) and long (30-day) microgravity experiments, we observed significant loss of bone tissue and structure in both the pelvis and the femur. After 15-days of microgravity on STS-131, pelvic ischium displayed a 6.23% decrease in bone fraction (p=0.005) and 11.91% decrease in bone thickness (p=0.002). Furthermore, during long-duration spaceflight we observed onset of an accelerated aging-like phenotype and osteoarthritic disease state indicating that stem cells within the bone tissue fail to repair and regenerate tissues in a normal manner, leading to drastic tissue alterations in response to microgravity. The Rodent Research Hardware System provides the capability to investigate these effects during long-duration experiments on the International Space Station. During the Rodent Research-1 mission 10 16-week-old female C57Bl/6J mice were exposed to 37-days of microgravity. All flight animals were euthanized and frozen on orbit for future dissection. Ground (n=10) and vivarium controls (n=10) were housed and processed to match the flight animal timeline. During this study we collected pelvis, femur, and tibia from all animal groups to test the hypothesis that stem cell-based tissue regeneration is significantly altered after 37-days of spaceflight. To do this, we will analyze differences in bone morphometric parameters using MicroCT. The pelvis, femur, and tibia are key in supporting and distributing weight under normal conditions. Therefore, we expect to see altered remodeling in flight animals in response to microgravity with respect to ground controls. In combination with histomorphometry, these results will help elucidate the complex mechanisms underlying bone tissue maintenance and stem cell regeneration.

microgravity↗

Ionizing Radiation Affects Gene Expression in Mouse Skin and Bone

Future long-duration space exploration beyond low earth orbit will increase human exposure to space radiation and microgravity conditions as well as associated risks to skeletal health. In animal studies, radiation exposure (greater than 1 Gy) is associated with pathological changes in bone structure, enhanced bone resorption, reduced bone formation and decreased bone mineral density, which can lead to skeletal fragility. Definitive measurements and detection of bone loss typically require large and specialized equipment which can make their application to long duration space missions logistically challenging. Towards the goal of developing non-invasive and less complicated monitoring methods to predict astronauts' health during spaceflight, we examined whether radiation induced gene expression changes in skin may be predictive of the responses of skeletal tissue to radiation exposure. We examined oxidative stress and growth arrest pathways in mouse skin and long bones by measuring gene expression levels via quantitative polymerase chain reaction (qPCR) after exposure to total body irradiation (IR). To investigate the effects of irradiation on gene expression, we used skin and femora (cortical shaft) from the following treatment groups: control (normally loaded, sham-irradiated), and IR (0.5 Gy 56Fe 600 MeV/n and 0.5 Gy 1H 150 MeV/n), euthanized at one and 11 days post-irradiation (IR). To determine the extent of bone loss, tibiae were harvested and cancellous microarchitecture in the proximal tibia quantified ex vivo using microcomputed tomography (microCT). Statistical analysis was performed using Student's t-test. At one day post-IR, expression of FGF18 in skin was significantly greater (3.8X) than sham-irradiated controls, but did not differ at 11 days post IR. Expression levels of other genes associated with antioxidant response (Nfe2l2, FoxO3 and Sod1) and the cell cycle (Trp53, Cdkn1a, Gadd45g) did not significantly differ between the control and IR groups at either time point. Radiation exposure resulted in a 27.0% increase in FGF18-positive hair follicles at one day post-IR and returned to basal levels at 11 days post-IR. A similar trend was observed from FGF18 gene expression analysis of skin. In bone (femora), there was an increase in the expression of the pro-osteoclastogenic cytokine, MCP-1, one day after IR compared to non-irradiated controls. FGF18 expression in skin and MCP- 1 expression in bone were found to be positively correlated (P less than 0.002, r=0.8779). Further, microcomputed tomography analysis of tibia from these animals showed reduced cancellous bone volume (-9.9%) at 11 days post- IR. These results suggest that measurements of early radiation induced changes in FGF18 gene expression in skin may have value for predicting subsequent loss of cancellous bone mass. Further research may lead to the development of a relatively simple diagnostic tool for bone loss, with the advantage that hair follicles and skin are relatively easy to acquire from human subjects.

space radiation↗

CDKN1a/p21 Plays a Critical Role in Suppressing Stem Cell Regenerative Potential During Aging

Unloading during spaceflight is known to adversely affect mammalian physiology. Mechanical stimulation is required for repair and regeneration by stem cell lineages to maintain tissue health and mass. CDKN1a/p21 functions as a potent cell cycle arrest molecule and we previously found that CDKN1a/p21 was overexpressed in mouse bone during 15-days of spaceflight on STS-131 and localized to osteoprecursor cells in the femur. Therefore, we hypothesized that altered expression of CDKN1a/p21 leads to an arrest of bone formation during spaceflight in response to altered load. To study CDKN1a/p21 and its role in stem cell-based tissue regeneration, we use a CDKN1a/p21 knockout (KO) mouse to investigate the impact on bone structure, osteoprogenitor proliferation, and mineralized nodule formation. We have shown that bone marrow stem cells isolated from juvenile (11-week-old) and skeletally mature (18-week-old) KO mice have an increased bone formation potential as evidenced by increased proliferation and mineralization rates. In addition, we have shown that juvenile KO mice display significantly increased bone volume fraction (BV/TV) relative to wildtype (WT) mice, but not in skeletally mature KO mice, indicating increased resorption and bone turnover in adult mice. To more closely examine age differences in the KO mouse, we will study a wider spectrum of mice ranging from 4 weeks to 12 months in age. To do this, we will analyze differences in bone morphometric parameters using MicroCT and osteoblastogenesis assays. The pelvis, femur, and tibia are key in distributing weight and we expect to see altered remodeling and stem cell potential with age. In combination with histomorphometry, these results will help elucidate the complex mechanisms underlying bone tissue maintenance and stem cell regeneration.

stem cells↗

Irradiation damage and IASCC of printed 316L for use as fuel cladding

Additive manufacturing is an area of increasing interest to the nuclear industry. It offers a number of benefits over traditional manufacturing methods, such as the ability to create complex geometries without the need for welds. In addition to the potential for creating nuclear structural components, additive manufacturing may be used to create novel fuel/cladding designs. However, printed steel behaves differently than traditional wrought material. A number of industries are already studying the effects of the printed microstructures on mechanical properties. For cladding material and some structural nuclear components, however, the nuclear industry must also understand the material response to irradiation. This work examines the irradiation damage and the irradiation assisted stress corrosion cracking behavior of both proton irradiated (1 and 5 dpa) printed 316L steel and wrought 316L steel that were irradiated side by side at the Michigan Ion Beam Laboratory. As orientation with respect to the build direction can affect the mechanical properties of printed metal, specimens were taken from multiple orientations within the printed steel. For the IASCC testing, specimens were strained to 4% in simulated BWR NWC water (288°C, 0.2 µS/cm). TEM, SEM and X-ray microCT were used to examine coupons of the material as well as tensile bars used in the IASCC testing. This paper will show the key findings from this work, namely the IASCC response of printed metal (as compared to wrought), as well as the effects of the printed microstructure (such as the pre-existing voids) on the irradiation damage.

36 MATERIALS SCIENCE↗

A Comparative Study of Compressive Effects on the Morphology and Performance of Carbon Paper and Cloth Electrodes in Redox Flow Batteries

Compressing porous carbon electrodes is a common approach to improve flow battery performance, but the resulting impact on electrode structure, fluid dynamics, and cell performance is not well understood. Herein, microtomographic imaging, load cell testing, and flow cell diagnostics are employed to characterize how compression‐induced changes impact pressure drop, polarization, and mass‐transfer scaling. Five different compressions are tested, spanning ranges typically used in literature, for AvCarb 1071 cloth (0%, 9%, 20%, 25%, 32%) and Freudenberg H23 paper (0%, 8%, 12%, 17%, 22%). It is found that the two electrode structures have distinct responses to compression, resulting in differing optimal conditions identified for each material; specifically, the Freudenberg H23 exhibits lower combined ohmic, charge‐transfer, and mass‐transport values at 8% compression, resulting in improved electrochemical performance across all compressive values, as compared to the optimal AvCarb 1071 compression (20%). Overall, Freudenberg H23 exhibits a greater sensitivity to compression with peak electrochemical activity correlating with increased permeability, whereas AvCarb 1071 is insensitive to compressive loads but produces lower electrochemical performance. Herein, the trade‐offs of mechanical robustness on fluid‐dynamic and electrochemical performance between the two electrodes are demonstrated by the aforementioned findings, suggesting each could be used for specific operating environments.

25 ENERGY STORAGE↗

The Wormholes Within: A Study of 1, 3, 5–Triamino–2, 4, 6–Trinitrobenzene Crystal Morphology by Micro and Nano–Scale X–Ray Computed Tomography and its Effects Upon the Shock Sensitivity of Detonation

We report that we understand heterogeneous-explosive detonation only to the extent that we can first measure the salient chemical and microstructural features. Barring an inordinate amount of trial and error, we can tailor the detonation properties of our HE formulations only to the extent that we understand (at least qualitatively) the same effects. As an insensitive high explosive, 1, 3, 5-triamino-2, 4, 6-trinitrobenzene (TATB) burns more heterogeneously than conventional high explosives, making its detonation properties sensitive to microstructure. The Benziger route can produce various grain morphologies depending on the amount of water used in the amination step. So-called dry-aminated (DA) TATB crystals are riddled with a void structure called wormholes, which are washed out ammonium chloride (NH4Cl) inclusions that form during synthesis. So-called wet-aminated (WA) TATB crystals do not contain NH 4 Cl inclusions but possess a very convoluted external structure. Using both micro and nano-scale CT, we explore three DA-TATB lots that reflect evolutionary processing changes, plus one WA-TATB material. Micro CT imaged crystal collections show the exterior grain structure, revealing its roughness and providing a crude particle size measurement. The DA-TATB lots that test most sensitively in the LANL ECOT corner-turning test are roughest, having the appearance of being “corroded”. Nano-scale CT reveals both internal NH 4 Cl inclusions and void structures. The DA-TATB lots that tested intermediate and high sensitivity in ECOT exhibited similar wormhole structures, which were more extensive than that of the low-sensitivity lot. The least sensitive DA-TATB lot also retained more NH 4 Cl, consistent with reduced connectivity of voids to the surface.

1, 3, 5-triamino-2,4↗

Interactive volumetric segmentation for textile micro-tomography data using wavelets and nonlocal means

This work addresses segmentation of volumetric images of woven carbon fiber textiles from micro-tomography data. We propose a semi-supervised algorithm to classify carbon fibers that requires sparse input as opposed to completely labeled images. The main contributions are: (a) design of effective discriminative classifiers, for three-dimensional textile samples, trained on wavelet features for segmentation; (b) coupling of previous step with nonlocal means as simple, efficient alternative to the Potts model; and (c) demonstration of reuse of classifier to diverse samples containing similar content. We evaluate our work by curating test sets of voxels in the absence of a complete ground truth mask. The algorithm obtains an average 0.95 F1 score on test sets and average F1 score of 0.93 on new samples. Finally, we conclude with discussion of failure cases and propose future directions toward analysis of spatiotemporal high-resolution micro-tomography images.

3D image processing↗

Non-invasive imaging reveals convergence in root and stem vulnerability to cavitation across five tree species

Root vulnerability to cavitation is challenging to measure and under-represented in current datasets. This gap limits the precision of models used to predict plant responses to drought because roots comprise the critical interface between plant and soil. In this study, we measured vulnerability to drought-induced cavitation in woody roots and stems of five tree species (Acacia aneura, Cedrus deodara, Eucalyptus crebra, Eucalytus saligna, and Quercus palustris) with a wide range of xylem anatomies. X-ray microtomography was used to visualize the accumulation of xylem embolism in stems and roots of intact plants that were naturally dehydrated to varying levels of water stress. Vulnerability to cavitation, defined as the water potential causing a 50% loss of hydraulic function (P 50 ), varied broadly among the species (–4.51 MPa to –11.93 MPa in stems and –3.13 MPa to –9.64 MPa in roots). The P 50 of roots and stems was significantly related across species, with species that had more vulnerable stems also having more vulnerable roots. While there was strong convergence in root and stem vulnerability to cavitation, the P 50 of roots was significantly higher than the P 50 of stems in three species. However, the difference in root and stem vulnerability for these species was small; between 1% and 31% of stem P 50 . Thus, while some differences existed between organs, roots were not dramatically more vulnerable to embolism than stems, and the differences observed were less than those reported in previous studies. Further study is required to evaluate the vulnerability across root orders and to extend these conclusions to a greater number of species and xylem functional types.

59 BASIC BIOLOGICAL SCIENCES↗

Structural organization of the spongy mesophyll

Summary Many plant leaves have two layers of photosynthetic tissue: the palisade and spongy mesophyll. Whereas palisade mesophyll consists of tightly packed columnar cells, the structure of spongy mesophyll is not well characterized and often treated as a random assemblage of irregularly shaped cells. Using micro‐computed tomography imaging, topological analysis, and a comparative physiological framework, we examined the structure of the spongy mesophyll in 40 species from 30 genera with laminar leaves and reticulate venation. A spectrum of spongy mesophyll diversity encompassed two dominant phenotypes: first, an ordered, honeycomblike tissue structure that emerged from the spatial coordination of multilobed cells, conforming to the physical principles of Euler’s law; and second, a less‐ordered, isotropic network of cells. Phenotypic variation was associated with transitions in cell size, cell packing density, mesophyll surface‐area‐to‐volume ratio, vein density, and maximum photosynthetic rate. These results show that simple principles may govern the organization and scaling of the spongy mesophyll in many plants and demonstrate the presence of structural patterns associated with leaf function. This improved understanding of mesophyll anatomy provides new opportunities for spatially explicit analyses of leaf development, physiology, and biomechanics.

3D↗

Interpreting Morphological Adaptations Associated with Viviparity in the Tsetse Fly Glossina morsitans (Westwood) by Three-Dimensional Analysis

Tsetse flies (genus Glossina), the sole vectors of African trypanosomiasis, are distinct from most other insects, due to dramatic morphological and physiological adaptations required to support their unique biology. These adaptations are driven by demands associated with obligate hematophagy and viviparous reproduction. Obligate viviparity entails intrauterine larval development and the provision of maternal nutrients for the developing larvae. The reduced reproductive capacity/rate associated with this biology results in increased inter- and intra-sexual competition. Here, we use phase contrast microcomputed tomography (pcMicroCT) to analyze morphological adaptations associated with viviparous biology. These include (1) modifications facilitating abdominal distention required during blood feeding and pregnancy, (2) abdominal and uterine musculature adaptations for gestation and parturition of developed larvae, (3) reduced ovarian structure and capacity, (4) structural features of the male-derived spermatophore optimizing semen/sperm delivery and inhibition of insemination by competing males and (5) structural features of the milk gland facilitating nutrient incorporation and transfer into the uterus. Three-dimensional analysis of these features provides unprecedented opportunities for examination and discovery of internal morphological features not possible with traditional microscopy techniques and provides new opportunities for comparative morphological analyses over time and between species.

59 BASIC BIOLOGICAL SCIENCES↗

Characterization of Incremental Markings in the Sagittal Otolith of the Pacific Sardine ( Sardinops sagax ) Using Different Imaging Modalities

Teleost fish possess calcium carbonate otoliths located in separate chambers (utriculus, sacculus, and lagena) of their membranous labyrinth. This study analyzed the surface topography of the sagittal otolith of the Pacific sardine (Sardinops sagax) and the daily and annual increments in these otoliths. The otolith surface, characterized by laser scanning confocal microscopy for the first time, consisted of a system of prominent ridges and valleys (grooves), but it is unclear whether these structures are functional or represent time-resolving markings reflecting growth periodicity. Within the first-year volume, daily increments, each consisting of an incremental (more mineralized) and a discontinuous (less mineralized) zone, were resolved by optical microscopy and backscattered electron (BSE) imaging in the scanning electron microscope (SEM). Daily growth increments could, however, not be resolved in volumes formed after the first year, presumably because otolith growth markedly slows down and spacing of incremental markings narrows in older fish. Throughout otolith growth, the crystalline network continues across the discontinuous zones. Fluorochrome labeling provides additional information on growth after the first year. Compared with optical and BSE imaging, synchrotron microComputed Tomography of intact otoliths (with 0.69 µm volume elements) was less able to resolve daily increments; X-ray phase contrast reconstructions provided more detail than reconstructions with absorption contrast. Future research directions are proposed.

59 BASIC BIOLOGICAL SCIENCES↗

37-Day Microgravity Exposure in 16-Week Female C57BL/6J Mice is Associated with Weight Bearing Site-Specific Bone Loss

Exposure to space environment factors, including microgravity unloading and space radiation, is well-known to be associated with rapid bone loss in mammals, but significant questions remain about mechanistic aspects of this process. Specifically, bone loss may vary depending on many interacting factors including age and skeletal maturity, physical activity, social versus individual housing, and habitat configuration. Likewise cellular tissue loss mechanisms in space may include changes to mineral homeostasis by activation of bone degradation by osteoclasts, or inhibition of bone formation by osteoblasts, and be caused by microgravity mechanical unloading or increased space radiation. In this study we used microcomputed tomography to quantify bone loss in multiple skeletal sites of group housed mice either exposed to 37 days in space on ISS during the Rodent Research 1, (RR1), experiments using the NASA Rodent Habitat, (RH), versus ground controls in the same habitats or standard vivarium cages. Our results in space versus matched RH ground controls show significant cancellous and cortical bone loss in the femur and tibia, but not in vertebrae, indicating that only weight-bearing bones experience significant bone loss during spaceflight. This finding suggests that space radiation at the low levels found in low Earth Orbit may not be a significant contributing factor to bone loss in space. Interestingly while mice maintained in the RH maintained or gained bone relative to Basal controls, mice kept in standard vivarium cages for 37 days showed significant loss in weight bearing bones, not as acute, but comparable in magnitude to spaceflight effects. These results suggest the RH configuration versus vivarium cages, may allow for greater mechanical loading of weight bearing bones as mice climb internal habitat surfaces, promoting maintenance of bone regenerative health.

mouse↗

Towards Quantifying and Correlating RTV Intumescence to Entry-Relevant Conditions

Room Temperature Vulcanizing silicone (RTV) is a high-temperature adhesive used as a gap-filler between Thermal Protection System (TPS) tiles for heatshields on numerous missions. It is also adopted to bond instrumentation plugs of temperature and pressure sensors into heatshield’s tiles. While RTV has been traditionally assumed to be a non-porous and non-ablating material, numerous experiments have shown that RTV pyrolyzes and becomes highly porous as it is heated. Experimental data has also shown heating rate-dependent swelling (or intumescence) and shrinking of RTV, these volume changes may cause roughness-induced boundary layer transition. Outgassing and surface oxide formation of RTV upon decomposition also occur, which can be sources of contamination of heat shield sensors. This combination of detrimental effects motivates a critical need to develop a high-fidelity model for RTV ablation. As a first critical step in RTV modeling, a comprehensive material properties database for RTV was collected that account for the lack of data in properties such as pyrolysis char yield, shape change, virgin and char porosity. The next step is to quantify the volume change of RTV as a function of temperature. While this was done in previous campaigns, limited data was collected which showed a possible heating rate dependent intumescent behavior. To further understand the intumescence phenomenon of RTV, numerous dedicated experiments were performed at Beamline 8.3.2 of the Advanced Light Source, where RTV samples were heated while X-ray scans were taken in situ. Micro-Computed Tomography scans were taken in situ for low heating rates (≤ 60 °C/min), whereas continuous radiography scans were taken for higher heating rates (≤ 1500 ºC/min). Unconstrained samples and samples constrained in different holders (quartz, graphite, FiberForm and PICA) were tested to determine the change in intumescence due to confinement. Using deep learning techniques, the tomographies and radiographies are segmented to obtain volume change, porosity, pore size distributions and connectivity. From the results obtained, modifications to the previous RTV intumescence model are proposed, along with preliminary comparisons to testing RTV at high-enthalpy facilities.

RTV↗