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The PAWS and STEM reliability analysis programs

The PAWS and STEM programs are new design/validation tools. These programs provide a flexible, user-friendly, language-based interface for the input of Markov models describing the behavior of fault-tolerant computer systems. These programs produce exact solutions of the probability of system failure and provide a conservative estimate of the number of significant digits in the solution. PAWS uses a Pade approximation as a solution technique; STEM uses a Taylor series as a solution technique. Both programs have the capability to solve numerically stiff models. PAWS and STEM possess complementary properties with regard to their input space; and, an additional strength of these programs is that they accept input compatible with the SURE program. If used in conjunction with SURE, PAWS and STEM provide a powerful suite of programs to analyze the reliability of fault-tolerant computer systems.

Butler, Ricky W.

Vacuum Ultraviolet Radiation and Atomic Oxygen Durability Evaluation of HST Bi-Stem Thermal Shield Materials

Bellows-type thermal shields were used on the bi-stems of replacement solar arrays installed on the Hubble Space Telescope (HST) during the first HST servicing mission (SMI) in December 1993. These thermal shields helped reduce the problem of thermal gradient- induced jitter observed with the original HST solar arrays during orbital thermal cycling and have been in use on HST for eight years. This paper describes ground testing of the candidate solar array bi-stem thermal shield materials including backside aluminized Teflon(R)FEP (fluorinated ethylene propylene) with and without atomic oxygen (AO) and ultraviolet radiation protective surface coatings for durability to AO and combined AO and vacuum ultraviolet (VOV) radiation. NASA Glenn Research Center (GRC) conducted VUV and AO exposures of samples of candidate thermal shield materials at HST operational temperatures and pre- and post-exposure analyses as part of an overall program coordinated by NASA Goddard Space Flight Center (GSFC) to determine the on-orbit durability of these materials. Coating adhesion problems were observed for samples having the AO- and combined AO/UV-protective coatings. Coating lamination occurred with rapid thermal cycling testing which simulated orbital thermal cycling. This lack of adhesion caused production of coating flakes from the material that would have posed a serious risk to HST optics if the coated materials were used for the bi-stem thermal shields. No serious degradation was observed for the uncoated aluminized Teflon(R) as evaluated by optical microscopy, although atomic force microscopy (AFM) microhardness testing revealed that an embrittled surface layer formed on the uncoated Teflon(R) surface due to vacuum ultraviolet radiation exposure. This embrittled layer was not completely removed by AO erosion, No cracks or particle flakes were produced for the embrittled uncoated material upon exposure to VUV and AO at operational temperatures to an equivalent exposure of approximately five years in the HST environment. Uncoated aluminized FEP Teflon(R) was determined to be the most appropriate thermal shield material and was used on the bi-stems of replacement solar arrays installed on HST during SMI in December 1993. The SMI -installed solar arrays air scheduled to be replaced during MST's fourth servicing mission (SM3B) in early 2002.

Dever, Joyce

Physical basis for altered stem elongation rates in internode length mutants of Pisum

Biophysical parameters related to gibberellin (GA)-dependent stem elongation were examined in dark-grown stem-length genotypes of Pisum sativum L. The rate of internode expansion in these genotypes is altered due to recessive mutations which affect either the endogenous levels of, or response to, GA. The GA deficient dwarf L181 (ls), two GA insensitive semierectoides dwarfs NGB5865 and NGB5862 (lka and lkb, respectively) and the slender' line L197 (la crys), which is tall regardless of GA content, were compared to the wild-type tall cultivar, Torsdag. Osmotic pressure, estimated by vapor pressure osmometry, and turgor pressure, measured directly with a pressure probe, did not correlate with the differences in growth rate among the genotypes. Mechanical wall properties of frozen-thawed tissue were measured using a constant force assay. GA deficiency resulted in increased wall stiffness judged both on the basis of plastic compliance and plastic extensibility normalized for equal stem circumference. Plastic compliance was not reduced in the GA insensitive dwarfs, though lka reduced circumference-normalized plasticity. In contrast, in vivo wall relaxation, determined by the pressure-block technique, differed among genotypes in a manner which did correlate with extension rates. The wall yield threshold was 1 bar or less in the tall lines, but ranged from 3 to 6 bars in the dwarf genotypes. The results with the ls mutant indicate that GA enhances stem elongation by both decreasing the wall yield threshold and increasing the wall yield coefficient. In the GA-insensitive mutants, lka and lkb, the wall yield threshold is substantially elevated. Plants possessing lka may also possess a reduced wall yield coefficient.

NASA Discipline Number 29-20

Mechanism of gibberellin-dependent stem elongation in peas

Stem elongation in peas (Pisum sativum L.) is under partial control by gibberellins, yet the mechanism of such control is uncertain. In this study, we examined the cellular and physical properties that govern stem elongation, to determine how gibberellins influence pea stem growth. Stem elongation of etiolated seedlings was retarded with uniconozol, a gibberellin synthesis inhibitor, and the growth retardation was reversed by exogenous gibberellin. Using the pressure probe and vapor pressure osmometry, we found little effect of uniconozol and gibberellin on cell turgor pressure or osmotic pressure. In contrast, these treatments had major effects on in vivo stress relaxation, measured by turgor relaxation and pressure-block techniques. Uniconozol-treated plants exhibited reduced wall relaxation (both initial rate and total amount). The results show that growth retardation is effected via a reduction in the wall yield coefficient and an increase in the yield threshold. These effects were largely reversed by exogenous gibberellin. When we measured the mechanical characteristics of the wall by stress/strain (Instron) analysis, we found only minor effects of uniconozol and gibberellin on the plastic compliance. This observation indicates that these agents did not alter wall expansion through effects on the mechanical (viscoelastic) properties of the wall. Our results suggest that wall expansion in peas is better viewed as a chemorheological, rather than a viscoelastic, process.

NASA Discipline Plant Biology

Long-term Blue Light Effects on the Histology of Lettuce and Soybean Leaves and Stems

Blue light (320 to 496 nm) alters hypocotyl and stem elongation and leaf expansion in short-term, cell-level experiments, but histological effects of blue light in long-term studies of whole plants have not been described. We measured cell size and number in stems of soybean (Glycine max L.) and leaves of soybean and lettuce (Lactuca sativa L.), at two blue light fractions. Short-term studies have shown that cell expansion in stems is rapidly inhibited when etiolated tissue is exposed to blue light. However, under long-term light exposure, an increase in the blue light fraction from less than 0.1% to 26% decreased internode length, specifically by inhibiting soybean cell division in stems. In contrast, an increase in blue light fraction from 6% to 26% reduced soybean leaf area by decreasing cell expansion. Surprisingly, lettuce leaf area increased with increasing blue light fraction (0% to 6%), which was attributed to a 3.1-fold increase in cell expansion and a 1.6-fold increase in cell division.

Dougher, Tracy A. O.

[STEM on Station Education]

The STEM on Station team is part of Education which is part of the External Relations organization (ERO). ERO has traditional goals based around BHAG (Big Hairy Audacious Goal). The BHAG model is simplified to a saying: Everything we do stimulates actions by others to advance human space exploration. The STEM on Station education initiate is a project focused on bringing off the earth research and learning into classrooms. Educational resources such as lesson plans, activities to connect with the space station and STEM related contests are available and hosted by the STEM on Station team along with their partners such as Texas Instruments. These educational activities engage teachers and students in the current happenings aboard the international space station, inspiring the next generation of space explorers.

Lundebjerg, Kristen

Stem Cell-Based Regenerative Health in Microgravity

The research of Dr. Almeida's Lab at NASA Ames focuses on the effects of microgravity mechanical unloading on stem-cell-based tissue regeneration. The research focuses on testing the broad hypothesis that mechanical load from gravity at 1 g is required for normal adult stem cell proliferation and differentiation during tissue regeneration. Research from the lab has provides evidence supporting the hypothesis in various biological models, including newt tail regeneration, mouse bone and bone marrow in whole animals, and mouse embryonic stem cells all exposed to microgravity during spaceflight. Using these models, the lab has identified overexpression of the cell cycle inhibitor p21/CDKN1a as a candidate molecular mediator mechanism of bone marrow mesenchymal and hematopoietic lineage tissue regenerative arrest in microgravity. The labis currently focused on using p21/CDKN1a null mouse and cell culture approaches in microgravity to test the gravity tissue regeneration hypothesis.

stem cells microgravity regeneration

Control of abdominal muscles by brain stem respiratory neurons in the cat

The nature of the control of abdominal muscles by the brain stem respiratory neurons was investigated in decerebrate unanesthetized cats. First, it was determined which of the brain stem respiratory neurons project to the lumbar cord (from which the abdominal muscles receive part of their innervation), by stimulating the neurons monopolarly. In a second part of the study, it was determined if lumbar-projecting respiratory neurons make monosynaptic connections with abdominal motoneurons; in these experiments, discriminate spontaneous spikes of antidromically acivated expiratory (E) neurons were used to trigger activity from both L1 and L2 nerves. A large projection was observed from E neurons in the caudal ventral respiratory group to the contralateral upper lumber cord. However, cross-correlation experiments found only two (out of 47 neuron pairs tested) strong monosynaptic connections between brain stem neurons and abdominal motoneurons.

Miller, Alan D.

The effectiveness of a double-stem injection valve in controlling combustion in a compression-ignition engine

An investigation was made to determine to what extent the rates of combustion in a compression-ignition engine can be controlled by varying the rates of fuel injection. The tests showed that the double-stem valve operated satisfactorily under all normal injection conditions; the rate of injection has a definite effect on the rate of combustion; the engine performance with the double-stem valve was inferior to that obtained with a single-stem valve; and the control of injection rates permitted by an injection valve of two stages of discharge is not sufficient to effect the desired rates of combustion.

Spanogle, J A

Development of Paper Products from Dried Sweetpotato Stems and Peanut Shells

One of the goals of NASA's Advanced Life Support Program (ALS) for sustaining human life in space is to achieve a closed system in plant production and usage. That all inedible plant parts should be recycled or used in some way. A Tuskegee University team researching sweetpotato and peanut for ALS has developed paper products from dried sweet-potato stems and peanut shells. In this study, the sweet-potato stems and peanut shells were soaked separately in water for 48 hours. After 48 hours, researchers manually separated the pulp and the unusable parts. To form the paper, 160 g of pulp and water mixture was poured through a 15.1 cm (diameter) filtration funnel and the pulp was trapped on 15 cm (diameter) filter paper. The filter paper and pulp were dried in an air oven, and the filter paper was removed, An examination under a scanning electron microscope showed that the sweet-potato paper was composed of "fibers", whereas the peanut shell paper was composed of "blocks". Results of physical testing showed that the sweet-potato stem paper was stronger than the peanut shell paper. It is anticipated that there may be other uses of these products such as writing paper, bags and packaging material. Because of its biodegradability, it can be incorporated into the resource recycling system at the end of its use.

R McConnell

The Hematopoietic Stem Cell Therapy for Exploration of Space

Astronauts experience severe/invasive disorders caused by space environments. These include hematological/cardiac abnormalities, bone and muscle losses, immunodeficiency, neurological disorders and cancer. While the cause of these symptoms are not yet fully delineated, one possible explanation could be the inhibition of hematopoietic stem cell (HSC) growth and hematopoiesis in space. HSCs differentiate into all types of blood cells, and growing evidence indicates that the HSCs also have the ability to transdifferentiate to various tissues, including muscle, skin, liver, neuronal cells and possibly bone. Therefore, a hypothesis was advanced in this laboratory that the hematopoietic stem cell-based therapy, herein called the hematopoietic stem cell therapy (HSCT), could mitigate some of the disorders described above. Due to the magnitude of this project our laboratory has subdivided it into 3 sections: a) HSCT for space anemia; b) HSCT for muscle and bone losses; and c) HSCT for immunodeficiency. Toward developing the HSCT protocol for space anemia, the HSC transplantation procedure was established using a mouse model of beta thalassemia. In addition, the NASA Rotating Wall Vessel (RWV) culture system was used to grow HSCs in space condition. To investigate the HSCT for muscle loss and bone loss, donor HSCs were genetically marked either by transfecting the beta-galactosidase-containing plasmid, pCMV.SPORT-beta-gal or by preparing from b-galactosidase transgenic mice. The transdifferentiation of HSCs to muscle is traced by the reporter gene expression in the hindlimb suspended mice with some positive outcome, as studied by the X-gal staining procedure. The possible structural contribution of HSCs against muscle loss is being investigated histochemically.

Roach, Allana Nicole

Cellular mechanisms underlying growth asymmetry during stem gravitropism

Plant stems respond to gravitropic stimulation with a rapid, local and reversible change in cell growth rate (elongation), generally on both the upper and lower sides of the stem. The cellular and biochemical mechanisms for this differential growth are reviewed. Considerable evidence implicates an asymmetry in wall pH in the growth response. The strengths and weaknesses of the wall "loosening enzyme" concept are reviewed and the possibility of expansin involvement in the bending response of stems is considered. Also discussed is the possibility that wall stiffening processes, e.g. phenolic coupling driven by oxidative bursts or altered orientation of newly deposited cellulose, might mediate the growth responses during gravitropism.

NASA Discipline Plant Biology

Kinetic separation of phototropism from blue-light inhibition of stem elongation

These experiments tested the hypothesis that phototropic bending arises when a light gradient across the stem differentially inhibits cell elongation because of direct inhibition of cell elongation by light (the Blaauw hypothesis). Continuous irradiation of dark-grown cucumber seedlings (Cucumis sativus L.) with unilateral blue light inhibited hypocotyl elongation within 30 s, but did not induce phototropic curvature until 4.5 h after the start of irradiation. Marking experiments showed that curvature began simultaneously at the top and bottom of the growing region. In situ measurements of the light gradient across the stem with a glass fiber optic indicated that a 5- to 6-fold difference in fluence rate was established on the two sides of the stem. The light gradient established at the start of irradiation was the same as that after 6 h of irradiation. Changes in gravitropic responsiveness during this period were also ruled out. Calculations show that the light gradient should have caused curvature which would be detectable within 30 to 60 min and which would extrapolate to the start of irradiation--if the Blaauw hypothesis were correct. The long lag for phototropism in this case indicates that rapid inhibition of cell elongation by blue light does not cause the asymmetrical growth of phototropism. Rather, phototropism is superimposed upon this separate light growth response.

NASA Discipline Number 40-30

Evaluation of Late Effects of Heavy-Ion Radiation on Mesenchymal Stem Cells

The overall objective of this recently funded study is to utilize well-characterized model test systems to assess the impact of pluripotent stem cell differentiation on biological effects associated with high-energy charged particle radiation. These stem cells, specifically mesenchymal stem cells (MSCs), have the potential for differentiation into bone, cartilage, fat, tendons, and other tissue types. The characterization of the regulation mechanisms of MSC differentiation to the osteoblastic lineage by transcription factors, such as Runx2/Cbfa1 and Osterix, and osteoinductive proteins such as members of the bone morphogenic protein family are well established. More importantly, for late biological effects, MSCs have been shown to contribute to tissue restructuring and repair after tissue injury. The complex regulation of and interactions between inflammation and repair determine the eventual outcome of the responses to tissue injury, for which MSCs play a crucial role. Additionally, MSCs have been shown to respond to reactive oxygen species, a secondary effector of radiation, by differentiating. With this, we hypothesized that differentiation of MSCs can alter or exacerbate the damage initiated by radiation, which can ultimately lead to late biological effects of misrepair/fibrosis which may ultimately lead to carcinogenesis. Currently, studies are underway to examine high-energy X-ray radiation at low and high doses, approximately 20 and 200 Rad, respectively, on cytogenetic damage and gene modulation of isolated MSCs. These cells, positive for MSC surface markers, were obtained from three persons. In vitro cell samples were harvested during cellular proliferation and after both cellular recovery and differentiation. Future work will use established in vitro models of increasing complexity to examine the value of traditional 2D tissue-culture techniques, and utilize 3D in vitro tissue culture techniques that can better assess late effects associated with radiation.

Gonda, S.R.

Extravehicular Activity Systems Education and Public Outreach in Support of NASA's STEM Initiatives in Fiscal Year 2011

NASA's goals to send humans beyond low Earth orbit will involve the need for a strong engineering workforce. Research indicates that student interest in science, technology, engineering, and math (STEM) areas is on the decline. According to the Department of Education, the United States President has mandated that 100,000 educators be trained in STEM over the next decade to reduce this trend. NASA has aligned its Education and Public Outreach (EPO) initiatives to include emphasis in promoting STEM. The Extravehicular Activity (EVA) Systems Project Office at the NASA Johnson Space Center actively supports this NASA initiative by providing subject matter experts and hands-on, interactive presentations to educate students, educators, and the general public about the design challenges encountered as NASA develops EVA hardware for exploration missions. This paper summarizes the EVA Systems EPO efforts and metrics from fiscal year 2011.

Paul, Heather L.

Extravehicular Activity Systems Education and Public Outreach in Support of NASA's STEM Initiatives in Fiscal Year 2011

NASA's goals to send humans beyond low Earth orbit will involve the need for a strong engineering workforce. Research indicates that student interest in science, technology, engineering, and math (STEM) areas is on the decline. According to the Department of Education, the United States President has mandated that 100,000 educators be trained in STEM over the next decade to reduce this trend. NASA has aligned its Education and Public Outreach (EPO) initiatives to include emphasis in promoting STEM. The Extravehicular Activity (EVA) Systems Project Office at the NASA Johnson Space Center actively supports this NASA initiative by providing subject matter experts and hands-on, interactive presentations to educate students, educators, and the general public about the design challenges encountered as NASA develops EVA hardware for exploration missions. This paper summarizes the EVA Systems EPO efforts and metrics from fiscal year 2011.

Paul, Heather

Single-Cell mRNA Sequencing of Bone Marrow Mesenchymal Stem Cell Lineages Shows that Gravity Mechanical Loading Regulates Regenerative Osteogenesis in a CDKN1A-Dependent Manner

Weight-bearing at 1g is increasingly recognized as an important stimulus driving stem cell-based tissue regenerative homeostasis and health. In the bone marrow compartment, mechanical stimuli from gravity can drive osteogenesis from mesenchymal progenitors and hematopoiesis from hematopoietic progenitors. Conversely, hematopoietic monocyte/macrophage progenitors can also be driven into osteoclastogenesis by mechanical unloading in microgravity. Although the cell cycle inhibitor gene Cdkn1a is known to be upregulated in microgravity, the great diversity of stem cell lineages and stages of differentiation in bone marrow, make it difficult to attribute gene expression changes to specific cell types. To overcome this limitation, we used single-cell mRNAseq analyses with both in-vitro and in-vivo strategies, including in-vitro stretch of marrow osteogenic cultures, and in-vivo hindlimb unloading, running wheels, treadmills, and exposure to real microgravity. For in-vitro studies marrow osteoprogenitors from WT and Cdkn1a-null mice were subjected to substrate stretch during osteogenesis, with static cultures simulating unloading. From those experiments we determined Cdkn1a-null cells transition more readily from progenitors to early osteoblasts and further to mineralizing osteoblasts. Additionally, single-cell mRNAseq resolution identified Cdkn1a suppression in response to mechanical loading predominantly in early proliferative osteoblasts rather than progenitor or mineralizing osteoblasts. These findings support the hypothesis that CDKN1A plays a mechano-reversible, anti-proliferative role during bone regenerative osteogenesis. To investigate in-vivo mechanoregulation of osteogenesis, mice were flown in microgravity during the Rodent Research-10 experiment comparing wildtype and Cdkn1a-null mice kept on ISS for 30 days. The mice were euthanized and dissected on-orbit in microgravity and femur bone marrow was collected using a custom sample isolation and live-cell cryo-preservation technique allowing for single cell transcriptomics post-flight processing. Ground control experiments with hindlimb-unloading, running wheels, and treadmills, were also conducted for a comprehensive analysis of mechanical loading effects on the regenerative health of bone tissue and its marrow stem cell progenitor lineages.

single cell sequencing

STEM Engagement and Robotics Workshops in the Community

It is my objective to inspire and engage students and the public in science, technology, engineering, and math (STEM) through hands on STEM activities during the City of Palmdale's Thursday Nights on the Square (TNOTS) as well as various robotics workshops throughout the AV and the San Bernardino County.

STEM