Leaf temperatures and environmental conditions predict daily stem radial variations in a temperate coniferous forest
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William A. Weygint↗
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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.
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Technical STEM Workforce Development Re-Imagined: The Lucy Student Pipeline and Competency Enabler (L’SPACE)
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Sheri Klug Boonstra↗
Telomere length assessments of muscle stem cells in rodent and human skeletal muscle sections
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Elisia D. Tichy↗
Structure of an anti-HIV-1 hammerhead ribozyme complex with a 17-mer DNA substrate analog of HIV-1 gag RNA and a mechanism for the cleavage reaction: 750 MHz NMR and computer experiments
The structure of an anti-HIV-1 ribozyme-DNA abortive substrate complex was investigated by 750 MHz NMR and computer modeling experiments. The ribozyme was a chimeric molecule with 30 residues-18 DNA nucleotides, and 12 RNA residues in the conserved core. The DNA substrate analog had 17 residues. The chimeric ribozyme and the DNA substrate formed a shortened ribozyme-abortive substrate complex of 47 nucleotides with two DNA stems (stems I and III) and a loop consisting of the conserved core residues. Circular dichroism spectra showed that the DNA stems assume A-family conformation at the NMR concentration and a temperature of 15 degrees C, contrary to the conventional wisdom that DNA duplexes in aqueous solution populate entirely in the B-form. It is proposed that the A-family RNA residues at the core expand the A-family initiated at the core into the DNA stems because of the large free energy requirement for the formation of A/B junctions. Assignments of the base H8/H6 protons and H1' of the 47 residues were made by a NOESY walk. In addition to the methyl groups of all T's, the imino resonances of stems I and III and AH2's were assigned from appropriate NOESY walks. The extracted NMR data along with available crystallographic data, were used to derive a structural model of the complex. Stems I and III of the final model displayed a remarkable similarity to the A form of DNA; in stem III, a GC base pair was found to be moving into the floor of the minor groove defined by flanking AT pairs; data suggest the formation of a buckled rhombic structure with the adjacent pair; in addition, the base pair at the interface of stem III and the loop region displayed deformed geometry. The loop with the catalytic core, and the immediate region of the stems displayed conformational multiplicity within the NMR time scale. A catalytic mechanism for ribozyme action based on the derived structure, and consistent with biochemical data in the literature, is proposed. The complex between the anti HIV-1 gag ribozyme and its abortive DNA substrate manifests in the detection of a continuous track of A.T base pairs; this suggests that the interaction between the ribozyme and its DNA substrate is stronger than the one observed in the case of the free ribozyme where the bases in stem I and stem III regions interact strongly with the ribozyme core region (Sarma, R. H., et al. FEBS Letters 375, 317-23, 1995). The complex formation provides certain guidelines in the design of suitable therapeutic ribozymes. If the residues in the ribozyme stem regions interact with the conserved core, it may either prevent or interfere with the formation of a catalytically active tertiary structure.
NASA Discipline Exobiology↗
A study of murine bone marrow cells cultured in bioreactors which create an environment which simulated microgravity
Previous research indicated that mouse bone marrow cells could be grown in conditions of simulated microgravity. This environment was created in rotating bioreactor vessels. On three attempts mouse cells were grown successfully in the vessels. The cells reached a stage where the concentrations were doubling daily. Phenotypic analysis using a panel of monoclonal antibodies indicated that the cell were hematopoietic pluripotent stem cells. One unsuccessful attempt was made to reestablish the immune system in immunocompromised mice using these cells. Since last summer, several unsuccessful attempts were made to duplicate these results. It was determined by electron microscopy that the cells successfully grown in 1989 contained virus particles. It was suggested that these virally parasitized cells had been immortalized. The work of this summer is a continuation of efforts to grow mouse bone marrow in these vessels. A number of variations of the protocol were introduced. Certified pathogen free mice were used in the repeat experiments. In some attempts the medium of last summer was used; in others Dexture Culture Medium containing Iscove's Medium supplemented with 20 percent horse serum and 10-6 M hydrocortisone. Efforts this summer were directed solely to repeating the work of last summer. Plans were made for investigations if stem cells were isolated. Immortalization of the undifferentiated stem cell would be attempted by transfection with an oncogenic vector. Selective differentiation would be induced in the stem cell line by growing it with known growth factors and immune response modulators. Interest is in identifying any surface antigens unique to stem cells that would help in their characterization. Another goal was to search for markers on stem cells that would distinguish them from stem cells committed to a particular lineage. If the undifferentiated hematopoietic stem cell was obtained, the pathways that would terminally convert it to myeloid, lyphoid, erythroid, or other cell lines would be studied. Transfection with a known gene would be attempted and then conversion to a terminally identifiable cell.
Lawless, Brother Desales↗
Cambium
The Cambium investigation is one in a pair of investigations which utilizes the Advanced Biological Research System (ABRS). Cambium seeks definitive evidence that gravity has a direct effect on cambial cells (cells located under the inner bark where secondary growth occurs) in willow, Salix babylonica. The Cambium investigation uses willow plants flown on the International Space Station to better understand the fundamental processes by which plants produce cellulose and lignin, the two main structural materials found in plant matter. On Earth, the nature of wood within tree stems varies depending on position, and that within-tree variation includes differences in cell types and chemistry including lignin and cellulose, two major components of wood influencing wood strength, usefulness and carbon content. Reaction wood is an extreme example of such variation, and it is believed that reaction wood develops as a reaction to the influence of gravity. For the Cambium experiment, young willow plants will be launched to the ISS where their stems will be looped in an attempt to induce reaction wood formation. After on-orbit growth, the plants will be preserved and returned to Earth for analysis. Understanding the role of gravity in wood formation is expected to enable wiser management of forests for carbon sequestration as well as better utilization of trees for wood products. Detailed Research Description: The Cambium experiment will provide an understanding of physiological processes such as gene expression, metabolism and general plant development that are affected in plant systems exposed to space flight. Cambium seeks definitive evidence that gravity has a direct effect on the cambial cells (cells located under the inner bark where secondary growth occurs) that contribute to xylogenesis (reaction wood formation) in willow plants, Salix babylonica. Tension wood fibers differentiate on the upper sides of stems when the stem is altered from its normal (vertical) growth position by looping. This reaction wood response does not occur if the orientation of the plant stem with respect to gravity is not altered. If a localized zone of tension wood should be formed in looped stems under microgravity conditions, this would be the first conclusive evidence that gravity is not required. On the other hand, if a zone of tension wood is not produced in looped stems (subjected to tension on one side, compression on the other) in microgravity, this would be the first definitive evidence that gravity has a direct effect on the cambial cells which contribute to reaction wood formation. Following return to Earth the plants will be analyzed by microscopy and chemical methods.
Savidge, Rodney↗
Gravitropism in higher plant shoots. IV - Further studies on participation of ethylene
Various hypotheses regarding the influence of ethylene on gravitropism in higher plant shoots were experimentally tested. It was found that ethylene at 1.0 and 10.0 cu cm/cu m decreased the rate of gravitropic bending in cocklebur stems, while 0.1 cm/cu m of ethylene had little effect. Treating cocklebur plants with 1.0 mmol aminoethoxyvinylglycine (AVG, an ethylene synthesis inhibitor) delayed stem bending compared with controls, but adding 0.1 cu cm/cu m ethylene in the surrounding atmosphere partially restored the rate of bending of AVG-treated plants. Virtually all newly synthesized ethylene appeared in bottom halves of horizontal stems, where ethylene concentrations were as much as 100 times those in upright stems or in top halves of horizontal stems. Auxin applied to one side of a vertical stem caused extreme bending away from that side; gibberellic acid, kinetin, and abscisic acid were without effect.
Wheeler, Raymond M.↗
Anatomical features of pepper plants (Capsicum annuum L.) grown under red light-emitting diodes supplemented with blue or far-red light
Pepper plants (Capsicum annuum L. cv., Hungarian Wax) were grown under metal halide (MH) lamps or light-emitting diode (LED) arrays with different spectra to determine the effects of light quality on plant anatomy of leaves and stems. One LED (660) array supplied 90% red light at 660 nm (25nm band-width at half-peak height) and 1% far-red light between 700-800nm. A second LED (660/735) array supplied 83% red light at 660nm and 17% far-red light at 735nm (25nm band-width at half-peak height). A third LED (660/blue) array supplied 98% red light at 660nm, 1% blue light between 350-550nm, and 1% far-red light between 700-800nm. Control plants were grown under broad spectrum metal halide lamps. Plants were gron at a mean photon flux (300-800nm) of 330 micromol m-2 s-1 under a 12 h day-night photoperiod. Significant anatomical changes in stem and leaf morphologies were observed in plants grown under the LED arrays compared to plants grown under the broad-spectrum MH lamp. Cross-sectional areas of pepper stems, thickness of secondary xylem, numbers of intraxylary phloem bundles in the periphery of stem pith tissues, leaf thickness, numbers of choloplasts per palisade mesophyll cell, and thickness of palisade and spongy mesophyll tissues were greatest in peppers grown under MH lamps, intermediate in plants grown under the 660/blue LED array, and lowest in peppers grown under the 660 or 660/735 LED arrays. Most anatomical features of pepper stems and leaves were similar among plants grown under 660 or 660/735 LED arrays. The effects of spectral quality on anatomical changes in stem and leaf tissues of peppers generally correlate to the amount of blue light present in the primary light source.
NASA Discipline Life Support Systems↗
Development of a Deep-Penetrating, Compact Geothermal Heat Flow System for Robotic Lunar Geophysical Missions
Geothermal heat flow measurements are a high priority for the future lunar geophysical network missions recommended by the latest Decadal Survey of the National Academy. Geothermal heat flow is obtained as a product of two separate measurements of geothermal gradient and thermal conductivity of the regolith/soil interval penetrated by the instrument. The Apollo 15 and 17 astronauts deployed their heat flow probes down to 1.4-m and 2.3-m depths, respectively, using a rotary-percussive drill. However, recent studies show that the heat flow instrument for a lunar mission should be capable of excavating a ~3-m deep hole to avoid the effect of potential long-term changes of the surface thermal environment. For a future robotic geophysical mission, a system that utilizes a rotary/percussive drill would far exceed the limited payload and power capacities of the lander/rover. Therefore, we are currently developing a more compact heat flow system that is capable of 3-m penetration. Because the grains of lunar regolith are cohesive and densely packed, the previously proposed lightweight, internal hammering systems (the so-called moles ) are not likely to achieve the desired deep penetration. The excavation system for our new heat flow instrumentation utilizes a stem which winds out of a pneumatically driven reel and pushes its conical tip into the regolith. Simultaneously, gas jets, emitted from the cone tip, loosen and blow away the soil. Lab tests have demonstrated that this proboscis system has much greater excavation capability than a mole-based heat flow system, while it weighs about the same. Thermal sensors are attached along the stem and at the tip of the penetrating cone. Thermal conductivity is measured at the cone tip with a short (1- to 1.5-cm long) needle sensor containing a resistance temperature detector (RTD) and a heater wire. When it is inserted into the soil, the heater is activated. Thermal conductivity of the soil is obtained from the rate of temperature increase during the heating. By stopping during the excavation, it is possible to measure thermal conductivities at different depths. The gas jets are turned off when the penetrating cone reaches the target depth. Then, the stem pushes the needle sensor into the undisturbed soil at the bottom of the hole and carries out a thermal conductivity measurement. When the measurement is complete, the system resumes excavation. RTDs, placed along the stem at short (approx 30 cm) intervals, will monitor long-term temperature stability of the subsurface. Temperature in the shallow subsurface would fluctuate with the diurnal, annual, and precession cycles of the Moon. These thermal waves penetrate to different depths into the regolith. Longterm monitoring of the subsurface temperature would allow us to accurately delineate these cyclic signals and separate them from the signal associated with the outward flow of the Moon s endogenic heat. Further, temperature toward bottom of the 3-m hole should be fairly stable after the heat generated during the excavation dissipates into the surrounding soil. The geothermal gradient may be determined reliably from temperature measurements at the RTDs near the bottom. In order to minimize the heat conduction along the stem from affecting the geothermal gradient measurements, we plan to use low-conductive materials for the stem and develop a mechanism to achieve close coupling between the RTDs and the wall of the excavated hole.
Nagihara, Seiichi↗
Inspiring the Next Generation of Explorers: Scientist Involvement in the Expedition Earth and Beyond Program
Scientists, science experts, graduate and even undergraduate student researchers have a unique ability to inspire the next generation of explorers. These science, technology, engineering, and mathematics (STEM) experts can serve as role models for students and can help inspire them to consider future STEM-related careers. They have an exceptional ability to instill a sense of curiosity and fascination in the minds of students as they bring science to life in the classroom. Students and teachers are hungry for opportunities to interact with scientists. They feel honored when these experts take time out of their busy day to share their science, their expertise, and their stories. The key for teachers is to be cognizant of opportunities to connect their students with scientists. For scientists, the key is to know how to get involved, to have options for participation that involve different levels of commitment, and to work with educational specialists who can help facilitate their involvement. The Expedition Earth and Beyond (EEAB) Program, facilitated by the Astromaterials Research and Exploration Science (ARES) Directorate at the NASA Johnson Space Center, is an Earth and planetary science education program designed to inspire, engage, and educate teachers and students by getting them actively involved with NASA exploration, discovery, and the process of science. One of the main goals of the program is to facilitate student research in the classroom. The program uses astronaut photographs, provided through the ARES Crew Earth Observations (CEO) payload on the International Space Station (ISS) as the hook to help students gain an interest in a research topic. Student investigations can focus on Earth or involve comparative planetology. Student teams are encouraged to use additional imagery and data from Earth or planetary orbital spacecraft, or ground-based data collection tools, to augment the astronaut photography dataset. A second goal of the program is to provide opportunities for meaningful connections between scientists and classrooms. To do this, EEAB offers multiple opportunities for scientist involvement. One opportunity involves having scientists work as mentors for student teams conducting research. These student teams, ranging from grades 4 through 12, are able to obtain guidance, suggestions, and input from STEM experts as they conduct a research investigation. Another opportunity for scientist involvement is participation in Classroom Connection Distance Learning (DL) events. These DL events entail interactive and engaging presentations that enable STEM experts to share their expertise with students and teachers (grades 3 through 12) from all across the nation. A third opportunity for scientist involvement involves participation in virtual student team science presentations. Student teams have the opportunity to share their research and results by presenting it to science experts through the use of WebEx, an easy-to-use online conferencing tool. The impact STEM experts have on students in today s classrooms is powerful. They serve as role models to these students, and they open students eyes to a potential career path they may not have known existed otherwise. The more scientists and STEM experts we can connect with students, the greater the impact we can make as we strive to inspire and prepare our nation s next generation of explorers.
Graff, Paige↗
NASA Space Science Day Events-Engaging Students in Science
The NASA Space Science Day Event follows the same format of planning and execution at all host universities and colleges. These institutions realized the importance of such an event and sought funding to continue hosting NSSD events. In 2014, NASA Johnson Space Center ARES team has supported the following universities and colleges that have hosted a NSSD event; the University of Texas at Brownsville, San Jacinto College, Georgia Tech University and Huston-Tillotson University. Other universities and colleges are continuing to conduct their own NSSD events. NASA Space Science Day Events are supported through continued funding through NASA Discovery Program. Community Night begins with a NASA speaker and Astromaterials display. The entire community surrounding the host university or college is invited to the Community Night. This year at the Huston-Tillotson (HTU) NSSD, we had Dr. Laurie Carrillo, a NASA Engineer, speak to the public and students. She answered questions, shared her experiences and career path. The speaker sets a tone of adventure and discovery for the NSSD event. After the speaker, the public is able to view Lunar and Meteorite samples and ask questions from the ARES team. The students and teachers from nearby schools attended the NSSD Event the following day. Students are able to see the university or college campus and the university or college mentors are available for questions. Students rotate through hour long Science Technology Engineering and Mathematics (STEM) sessions and a display area. These activities are from the Discovery Program activities that tie in directly with k- 12 instruction. The sessions highlight the STEM in exploration and discovery. The Lunar and Meteorite display is again available for students to view and ask questions. In the display area, there are also other interactive displays. Angela Green, from San Jacinto College, brought the Starlab for students to watch a planetarium exhibit for the NSSD at Huston-Tillotson University. Many HTU mentors were leading activities in the display room such as build a comet, volcano layering and robotics manipulation. Students were exposed to a variety STEM career possibilities and information. The students could relate the displays and sessions to what they were learning in school. The HTU mentors made the connection clear for the students. The students ended the event with a mission design presentation. They were able to take what they had learned during the day and were able to create a mission. Students presented their Mission Design and gained confidence in STEM. Conclusion: NASA Space Science Day Events provides an out of school experiential learning environment for students to enhance their STEM curriculum and let students see a college campus. The experiences students gain from attending NSSD gives them the confidence to see themselves on a college campus, possibly majoring in a STEM degree, and understand the importance of completing school.
Foxworth, S.↗
We Are All Potential Role Models: How We Can Support Scientist Engagement with Youth and the General Public
We all have the opportunity to be positive STEM role models. Yet scientists rarely identify themselves as a possible role model. In the training, “Empowering Effective STEM Role Models,” scientists practice research based strategies for engaging youth, particularly girls. This type of training is complementary to science communication, but is focused on equipping scientists with the skills needed to successfully encourage youth to pursue STEM careers. This training has been provided to over 150 STEM professionals at NASA and 14 federal agencies participated in a Train-the-Trainer version, organized by the White House Council for Women and Girls in STEM Working Group. Participants in this training reported that learning and practicing how to connect with and inspire children in STEM made them more confident stepping into further engagement opportunities. Confident, inspiring scientists are a highlight of a successful engagement event, but other factors make a positive experience possible.It’s important that all partners are thoughtful and intentional in preparing for public engagement events. The logistics are crucial, especially now while many engagements have moved to virtual platforms. Additional technology considerations make preparation essential. Two examples of virtual engagement highlight the support provided that led to successful engagements. The GLOBE Mission Earth project engages scientists in virtual classroom visits. To ensure both teacher and scientist expectations are met, a coordinator facilitates up front preparation by providing slide templates for the scientist to customize and providing video and other resources to the teacher and students to prompt student questions. Another example leverages the power of social media, particularly NASA Facebook. The NASA Earth Science Education Collaborative organizes citizen science challenges and engages the public on social. For one event, scientists were asked to share their connection to the investigation, but without access to NASA’s video production studio, the plans shifted. Scientists prepared audio files instead of videos and collaborated with communications leads to help craft their message. This weekly series has been flexible, adjusting to scientist's comfort levels, and has resulted in fresh and creative engagement.
Jessica Elaine Taylor↗
Femoral Head Bone Loss Following Short and Long-Duration Spaceflight
Exposure to mechanical unloading during spaceflight is known to have significant effects on the musculoskeletal system. Our ongoing studies with the mouse bone model have identified the failure of normal stem cell-based tissue regeneration, in addition to tissue degeneration, as a significant concern for long-duration spaceflight, especially in the mesenchymal and hematopoietic tissue lineages. The 30-day BionM1 and the 37-day Rodent Research 1 (RR1) missions enabled the possibility of studying these effects in long-duration microgravity experiments. We hypothesized that the inhibition of stem cell-based tissue regeneration in short-duration spaceflight would continue during long-duration spaceflight and furthermore would result in significant tissue alterations. MicroCT analysis of BionM1 femurs revealed 31 decrease in bone volume ratio, a 14 decrease in trabecular thickness, and a 20 decrease in trabecular number in the femoral head of space-flown mice. Furthermore, high-resolution MicroCT and immunohistochemical analysis of spaceflight tissues revealed a severe disruption of the epiphyseal boundary, resulting in endochondral ossification of the femoral head and perforation of articular cartilage by bone. This suggests that spaceflight in microgravity may cause rapid induction of an aging-like phenotype with signs of osteoarthritic disease in the hip joint. However, mice from RR1 exhibited significant bone loss in the femoral head but did not exhibit the severe aging and disease-like phenotype observed during BionM1. This may be due to increased physical activity in the RH hardware. Immunohistochemical analysis of the epiphyseal plate and investigation of cellular proliferation and differentiation pathways within the marrow compartment and whole bone tissue is currently being conducted to determine alterations in stem cell-based tissue regeneration between these experiments. Our results show that the observed inhibition of stem cell-based tissue regeneration persists during long-duration spaceflight. Furthermore, spaceflight femurs from BionM1 indicate onset of an accelerated aging-like phenotype with signs of osteoarthritic disease shown by disruption of the epiphyseal boundary and endochondral ossification. These effects are likely caused by a failure of stem cells to regenerate degraded tissues and may have significant implications for bone and cartilage health following extensive periods of mechanical unloading during long-duration spaceflight.
stem cells↗
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↗
Femoral Head Bone Loss Following Short and Long-Duration Spaceflight
Exposure to mechanical unloading during spaceflight is known to have significant effects on the musculoskeletal system. Our ongoing studies with the mouse bone model have identified the failure of normal stem cell-based tissue regeneration, in addition to tissue degeneration, as a significant concern for long-duration spaceflight, especially in the mesenchymal and hematopoietic tissue lineages. The 30-day BionM1 and the 37-day Rodent Research 1 (RR1) missions enabled the possibility of studying these effects in long-duration microgravity experiments. We hypothesized that the inhibition of stem cell-based tissue regeneration in short-duration spaceflight would continue during long-duration spaceflight and furthermore would result in significant tissue alterations. MicroCT analysis of BionM1 femurs revealed 31% decrease in bone volume ratio, a 14% decrease in trabecular thickness, and a 20% decrease in trabecular number in the femoral head of space-flown mice. Furthermore, high-resolution MicroCT and immunohistochemical analysis of spaceflight tissues revealed a severe disruption of the epiphyseal boundary, resulting in endochondral ossification of the femoral head and perforation of articular cartilage by bone. This suggests that spaceflight in microgravity may cause rapid induction of an aging-like phenotype with signs of osteoarthritic disease in the hip joint. However, mice from RR1 exhibited significant bone loss in the femoral head but did not exhibit the severe aging and disease-like phenotype observed during BionM1.This may be due to increased physical activity in the RH hardware. Immunohistochemical analysis of the epiphyseal plate and investigation of cellular proliferation and differentiation pathways within the marrow compartment and whole bone tissue is currently being conducted to determine alterations in stem cell-based tissue regeneration between these experiments. Our results show that the observed inhibition of stem cell-based tissue regeneration persists during long-duration spaceflight. Furthermore, spaceflight femurs from BionM1 indicate onset of an accelerated aging-like phenotype with signs of osteoarthritic disease shown by disruption of the epiphyseal boundary and endochondral ossification. These effects are likely caused by a failure of stem cells to regenerate degraded tissues and may have significant implications for bone and cartilage health following extensive periods of mechanical unloading during long-duration spaceflight.
microgravity↗
The Effects of CDKN1a/p21 on Oxidative Stress and Mitochondrial Function During Long Duration Spaceflight
Broad tissue degeneration and the failure of normal tissue regenerative processes in microgravity because of mechanical unloading are increasing concerns for sustaining life in space as the duration of future flight missions increases. Work in our laboratory has identified normal adult stem cell-based tissue regenerative processes, such as the formation of new bone, cartilage, and immune cells, as being particularly sensitive to the stresses of mechanical unloading in microgravity. Our studies have also identified the inhibition of differentiation of marrow mesenchymal stem cells and activation of CDKN1ap21-mediated cell cycle arrest in proliferative osteoprecursor cells on the bone surface as potential mechanisms for spaceflight-induced skeletal changes. This finding, in combination with the role of CDKN1ap21 as a suppressor of mammalian tissue regeneration, suggests that this gene could be responsible for suppressing stem cell-based tissue regeneration in response to disuse. In this work, we hypothesized that CDKN1ap21 regulates regenerative bone formation in response to alterations in mechanical load and tested this hypothesis by studying the skeletal phenotype and stem cell regenerative ability of juvenile (4-11 weeks old) and adult (18 weeks-12 months old) p21 (--) knockout (KO) mice. Additionally, we analyzed bone micro-architectural properties, bone formation rates and differentiation capacity of bone marrow stem cells (BMSCs) from male and female KO mice exposed to hindlimb unloading (HU) for 15-30 days. We found that juvenile KO mice exhibited increased femoral trabecular and cortical bone formation, whilst three-point bending of the tibias from KO mice showed decreased bone stiffness. Conversely, adult KO mice exhibited no significant differences in micro-architectural properties compared to WT (wild-type) but woven bone structure was indicative of rapid bone remodeling. Furthermore, cortical bone properties showed similar characteristics to aged bone, including increased cross-sectional area and perimeter, whilst three-point bending showed increased stiffness and toughness. Interestingly, in-vitro, KO mice exhibited increased differentiation and mineralized nodule formation in osteoblastogenesis assays compared to WT. Preliminary results from CDKN1ap21 KO mice subjected to HU suggest altered sensitivity to mechanical unloading resulting in decreased cortical thickness compared to WT mice. However, KO mice subjected to short and long-duration HU show increased in-vitro differentiation potential of BMSCs to from form mature, mineral-forming osteoblasts, indicating maintenance of regenerative potential. Analysis of bone formation rates, cell proliferation rates and key genes of interest are currently underway. These results indicate a novel role for CDKN1ap21 in load-dependent osteoprogenitor proliferation and differentiation and that deletion of CDKN1ap21 results in an age-dependent release of osteoblast proliferation inhibition and increased bone formation and turnover.
Stem Cells↗
Proximate nutritional composition of CELSS crops grown at different CO2 partial pressures
Two Controlled Ecological Life Support System (CELSS) candidate crops, soybean (Glycine max) and potato (Solanum tuberosum), were grown hydroponically in controlled environments maintained at carbon dioxide (CO2) partial pressures ranging from 0.05 to 1.00 kPa (500 to 10,000 ppm at 101 kPa atmospheric pressure). Plants were harvested at maturity (90 days for soybean and 105 days for potato) and all tissues analyzed for proximate nutritional composition (i.e. protein, fat, carbohydrate, crude fiber, and ash content). Soybean seed ash and crude fiber were higher and carbohydrate was lower than values reported for field-grown seed. Potato tubers showed little difference from field-grown tubers. Crude fiber of soybean stems and leaves increased with increased CO2, as did soybean leaf protein (total nitrogen). Potato leaf and stem (combined) protein levels also increased with increased CO2, while leaf and stem carbohydrates decreased. Values for leaf and stem protein and ash were higher than values generally reported for field-grown plants for both species. Results suggest that CO2 partial pressure should have little influence on proximate composition of potato tubers or soybean seed, but that high ash and protein levels might be expected from leaves and stems of crops grown in controlled environments of a CELSS.
Wheeler, R. M.↗
Thigmomorphogenesis: anatomical, morphological and mechanical analysis of genetically different sibs of Pinus taeda in response to mechanical perturbation
Twenty-three open pollinated families (half-sibs) and four controlled pollinated families (full-sibs) of Pinus taeda L. (loblolly pine) were grown in a greenhouse and analyzed for changes induced by mechanical perturbation (MP). These changes included inhibition of stem and needle elongation, bracing of branch nodes, and increased radial growth in the direction of the MP. Inhibition of stem elongation was the least variable feature measured. Leaf extension and stem diameter were highly variable between half-sibs. MP induced increased drag in greenhouse grown P. taeda in wind-tunnel tests. In P. taeda, MP induced decreased flexibility and increased elasticity and plasticity of the stem. The increased radial growth of the stems overrode the increase in elasticity, resulting in an overall decrease in flexibility. MP trees had a higher rupture point than non-MP controls. Increased radial growth is a result of more rapid cell divisions of the vascular cambium, resulting in increased numbers of tracheids. The decreased leader growth is partly due to a decreased tracheid length in response to MP.
Non-NASA Center↗