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At least 235 records · Page 13

Differential Gene Expression in A Cross-Feeding Two-Species Model Microbial Community Under Simulated Microgravity and Deep-Space Radiation

A long-term goal of space biology is to understand interspecies microbial interactions in space. Presently, little is known about the combined effect of microgravity and ionizing radiation on bacterial community response when species are interdependent through exchange of metabolites in fluid medium (cross-feeding). Microgravity is expected to slow interspecies mass transfer and growth in cross-feeding communities in the low-shear, diffusion-limited environment, while ionizing radiation may influence stress response to direct (DNA damage) and indirect damage (ROS). Using a well-understood, two-species (Escherichia coli and Salmonella enterica) microbial community engineered to be a model for studying cross-feeding, we simulated galactic cosmic rays (GCRsim) and microgravity to test the hypothesis: exposure to ionizing radiation causes cell damage or stress, altering transcriptomic community responses in metabolically interdependent cells, which is exacerbated by microgravity. We expect to see differential gene expression between cross-feeding and non-cross-feeding communities. We measured GCRsim effects on growth and gene expression in well-mixed versus simulated-microgravity conditions and in cross-feeding and non-cross-feeding medium. Microbial cultures were inoculated into liquid medium in rotating wall vessels (RWV) with different rotation rates: 5 RPM (simulated microgravity) and 50 RPM (well-mixed). The E. coli-S. enterica consortium, under simulated microgravity, were exposed to 500 mGy of Simplified 5-ion Galactic Cosmic Ray Simulation for 2 hours at Brookhaven National Lab. We harvested samples 40 minutes after irradiation for extraction and sequencing (NASA GeneLab). Here we present the differential gene expression analysis results, which reveal altered transcriptomic community responses, even where growth rate differences are not observed. Gene expression of these actively metabolizing microbial communities in GCRsim may illuminate molecular mechanisms of microbial interactions in space. Understanding how microbial community gene expression, metabolism, and other cellular processes are influenced by spaceflight stressors can inform the use of microbes in human life support for low Earth orbit missions and beyond.

microgravity↗

Low-shear modeled microgravity: a global environmental regulatory signal affecting bacterial gene expression, physiology, and pathogenesis

Bacteria inhabit an impressive variety of ecological niches and must adapt constantly to changing environmental conditions. While numerous environmental signals have been examined for their effect on bacteria, the effects of mechanical forces such as shear stress and gravity have only been investigated to a limited extent. However, several important studies have demonstrated a key role for the environmental signals of low shear and/or microgravity in the regulation of bacterial gene expression, physiology, and pathogenesis [Chem. Rec. 1 (2001) 333; Appl. Microbiol. Biotechnol. 54 (2000) 33; Appl. Environ. Microbiol. 63 (1997) 4090; J. Ind. Microbiol. 18 (1997) 22; Curr. Microbiol. 34(4) (1997) 199; Appl. Microbiol. Biotechnol. 56(3-4) (2001) 384; Infect Immun. 68(6) (2000) 3147; Cell 109(7) (2002) 913; Appl. Environ. Microbiol. 68(11) (2002) 5408; Proc. Natl. Acad. Sci. U. S. A. 99(21) (2002) 13807]. The response of bacteria to these environmental signals, which are similar to those encountered during prokaryotic life cycles, may provide insight into bacterial adaptations to physiologically relevant conditions. This review focuses on the current and potential future research trends aimed at understanding the effect of the mechanical forces of low shear and microgravity analogues on different bacterial parameters. In addition, this review also discusses the use of microgravity technology to generate physiologically relevant human tissue models for research in bacterial pathogenesis.

NASA Discipline Cell Biology↗

Mechanical behavior of carbon-carbon composites

A general background, test plan, and some results of preliminary examinations of a carbon-carbon composite material are presented with emphasis on mechanical testing and inspection techniques. Experience with testing and evaluation was gained through tests of a low modulus carbon-carbon material, K-Karb C. The properties examined are the density - 1.55 g/cc; four point flexure strength in the warp - 137 MPa (19,800 psi) and the fill - 95.1 MPa (13,800 psi,) directions; and the warp interlaminar shear strength - 14.5 MPa (2100 psi). Radiographic evaluation revealed thickness variations and the thinner areas of the composite were scrapped. The ultrasonic C-scan showed attenuation variations, but these did not correspond to any of the physical and mechanical properties measured. Based on these initial tests and a survey of the literature, a plan has been devised to examine the effect of stress on the oxidation behavior, and the strength degradation of coated carbon-carbon composites. This plan will focus on static fatigue tests in the four point flexure mode in an elevated temperature, oxidizing environment.

Rozak, G. A.↗

Matrix Fatigue Cracking Mechanisms of Alpha(2) TMC for Hypersonic Applications

The objective of this work was to understand matrix cracking mechanisms in a unidirectional alpha(sub 2) TMC in possible hypersonic applications. A (0)(sub 8) SCS-6/Ti-24Al-11Nb (at. percent) TMC was first subjected to a variety of simple isothermal and nonisothermal fatigue cycles to evaluate the damage mechanisms in simple conditions. A modified ascent mission cycle test was then performed to evaluate the combined effects of loading modes. This cycle mixes mechanical cycling at 150 and 483 C, sustained loads, and a slow thermal cycle to 815 C. At low cyclic stresses and strains more common in hypersonic applications, environment-assisted surface cracking limited fatigue resistance. This damage mechanism was most acute for out-of-phase nonisothermal cycles having extended cycle periods and the ascent mission cycle. A simple linear fraction damage model was employed to help understand this damage mechanism. Time-dependent environmental damage was found to strongly influence out-of-phase and mission life, with mechanical cycling damage due to the combination of external loading and CTE mismatch stresses playing a smaller role. The mechanical cycling and sustained loads in the mission cycle also had a smaller role.

Gabb, Timothy P.↗

Space environmental effects on polymer composites: Research needs and opportunities

The long-term performance of polymer-based composites in the space environment is discussed. Both thermoset and thermoplastic matrix composites are included in this discussion. Previous efforts on the space environmental effects on composites are briefly reviewed. Focus of this review is placed on the effects of hygrothermal stresses, atomic oxygen, ultraviolet (UV), and space debris/micrometeoroid impacts along with the potential synergism. Potential approaches to estimating the residual strength of polymer composites after exposure to atomic oxygen erosion or space debris/micrometeoroid impact are evaluated. New ground-based data are then utilized to illustrate the effects of atomic oxygen and thermal cycling on the failure behavior of polymer composites. Finally, research needs, challenges, and opportunities in the field of space environmental effects on composite materials are highlighted.

Jang, Bor Z.↗

Microbial Communities in Microgravity: Simulation in Lab and on the Computer

Microorganisms grow differently in spaceflight than they do on Earth. While much remains unexplained about how microgravity affects microbial growth, one dominant hypothesis is that the lack of density-driven convection in the liquid growth environment makes mixing diffusion-limited, and therefore slower. This is supported by evidence that individual microbial strains experience starvation and acid stress in microgravity. However, if it is true, then microgravity would also have measurable effects on microbes in mixed communities, because many interspecies interactions involve the exchange of soluble metabolites through the medium (cross-feeding). Specifically, cooperative cross-feeding communities would grow more slowly in microgravity, and cooperation would be less stable on evolutionary timescales. Here we describe our efforts to test this hypothesis by simulating microgravity in silico and in the lab, using a model system of Eschericia coli and Salmonella enterica that grow only when they can exchange methionine and acetate. We created CAMDLES (CFD-DEM Artificial Microgravity Developments for Living Ecosystem Simulation) as an extension of CFDEM®coupling software, to carry out computational modeling of biological flows, growth, and mass transfer in microgravity and also in laboratory artificial microgravity devices (rotating wall vessels, RWV). Using CAMDLES, we found distinct differences in growth rates between RWV and true microgravity, and we were able to identify several features, such as spatial distribution, biofilm formation, and product yield parameters, that influence the degree to which RWV growth recapitulates microgravity growth. In addition, we report on the development of a laboratory system for monitoring growth rates and species ratios of the community in RWVs, using fluorescent strains. Pairing CAMDLES with the laboratory model system allows us to generate quantitative predictions about the effects of spaceflight on organisms that will be essential to sustaining human space exploration in the long term.

microbiology↗

A mirror mount for cryogenic environments

The finite element method was used to study the effect of mount-induced aberrations on the optical surface of a lightweight double arch mirror subjected to cryogenic temperatures. The mount design was controlled by the requirements imposed on the optical surface quality and stress levels. The finite element analysis was used to define the feasible range of mount parameters and the selection of a design within the feasible region. The final design consisted of three spring-loaded Invar T-clamps that uniquely define the location of the mirror, three radially compliant parallel spring guides that remove the effect of radial contraction of structure in cryogenic temperatures, and a flexible baseplate that was used to reduce the effect of temperature-induced baseplate tilt errors. The experimental results from the application of this system to an existing 20-inch fused silica double arch mirror are shown, and possible improvements in system performance are discussed.

Iraninejad, B.↗

Secondary metabolism in simulated microgravity

We have studied microbial secondary metabolism in a simulated microgravity (SMG) environment provided by NASA rotating-wall bioreactors (RWBs). These reactors were designed to simulate some aspects of actual microgravity that occur in space. Growth and product formation were observed in SMG in all cases studied, i.e., Bacillus brevis produced gramicidin S (GS), Streptomyces clavuligerus made beta-lactam antibiotics, Streptomyces hygroscopicus produced rapamycin, and Escherichia coli produced microcin B17 (MccB17). Of these processes, only GS production was unaffected by SMG; production of the other three products was inhibited. This was determined by comparison with performance in an RWB positioned in a different mode to provide a normal gravity (NG) environment. Carbon source repression by glycerol of the GS process, as observed in shaken flasks, was not observed in the RWBs, whether operated in the SMG or NG mode. The same phenomenon occurred in the case of MccB17 production, with respect to glucose repression. Thus, the negative effects of carbon source on GS and beta-lactam formation are presumably dependent on shear, turbulence, and/or vessel geometry, but not on gravity. Stimulatory effects of phosphate and the precursor L-lysine on beta-lactam antibiotic production, as observed in flasks, also occurred in SMG. An almost complete shift in the localization of produced MccB17 from cells to extracellular medium was observed when E. coli was grown in the RWB under SMG or NG. If a plastic bead was placed in the RWB, accumulation became cellular, as it is in shaken flasks, indicating that sheer stress favors a cellular location. In the case of rapamycin, the same type of shift was observed, but it was less dramatic, i.e., growth in the RWB under SMG shifted the distribution of produced rapamycin from 2/3 cellular:1/3 extracellular to 1/3 cellular:2/3 extracellular. Stress has been shown to induce or promote secondary metabolism in a number of other microbial systems. RWBs provide a low stress SMG environment, which, however, supports only poor production of MccB17, as compared to production in shaken flasks. We wondered whether the poor production in RWBs under SMG is due to the low level of stress, and whether increasing stress in the RWBs would raise the amount of MccB17 formed. We found that increasing shear stress by adding a single Teflon bead to the RWB improved MccB17 production. Although shear stress seems to have a marked positive effect on MccB17 production in SMG, addition of various concentrations of ethanol to RWBs (or to shaken flasks) failed to increase MccB17 production. Ethanol stress merely decreased production and, at higher concentrations, inhibited growth. Interestingly, cells growing in the RWB were much more resistant to the growth- and production-inhibitory effects of ethanol than cells growing in shaken flasks. With respect to S. hygroscopicus, addition of Teflon beads to the RWB reversed the inhibition of growth, but rapamycin production was still markedly inhibited, and the distribution did not revert back to a preferential cellular site.

NASA Discipline Cell Biology↗

Investigation of flaw geometry and loading effects on plane strain fracture in metallic structures

The effects on fracture and flaw growth of weld-induced residual stresses, combined bending and tension stresses, and stress fields adjacent to circular holes in 2219-T87 aluminum and 5AI-2.5Sn(ELI) titanium alloys were evaluated. Static fracture tests were conducted in liquid nitrogen; fatigue tests were performed in room air, liquid nitrogen, and liquid hydrogen. Evaluation of results was based on linear elastic fracture mechanics concepts and was directed to improving existing methods of estimating minimum fracture strength and fatigue lives for pressurized structure in spacecraft and booster systems. Effects of specimen design in plane-strain fracture toughness testing were investigated. Four different specimen types were tested in room air, liquid nitrogen and liquid hydrogen environments using the aluminum and titanium alloys. Interferometry and holograph were used to measure crack-opening displacements in surface-flawed plexiglass test specimens. Comparisons were made between stress intensities calculated using displacement measurements, and approximate analytical solutions.

Hall, L. R.↗

Potting procedure for electronic components

Potting process is modified to effect a match more closely between embedded electronic components, potting mediums, and thermal environment. Application of room-temperature vulcanizing silicone rubber band cured in modified thermal cycle minimizes coil-to-resin adhesion and thus lowers stresses between transformer and potting compound.

Rubino, A. G.↗

Constitutive and Stability Behavior of Soils in Microgravity Environment

All aspects of soil stability, bearing capacity, slope stability, the supporting capacity of deep foundations, and penetration resistance depend on soil strength. The stress-deformation and stress-deformation-time behavior of soils are of importance in any problem where ground movements are of interest. In most engineering materials, the strength is derived from internal chemical and physico-chemical forces of interaction, which bond the atoms, molecules, and particles together. In soils, the constitutive relations are mainly derived from interparticle friction between particles and particle groups and dilatancy, and to a lesser extent from particle bonding by weak electrostatic, physico-chemical, and coulomb forces. For engineering purposes, soils are classified as cohesive (clays and silts; typical particle sizes range from 10 nm to 10 micrometers) and cohesionless (sand and gravel; typical particle sizes range from 10 micrometers to 75 mm). The mechanical or constitutive properties of cohesionless soils or granular materials are highly fabric-dependent, highly non-linear, and non-conservative with engineering properties primarily depending on the effects of gravity through self-weight and on the tractions or forces applied to the soil mass. Under moderate-to-high stress levels, the influence of gravity on the behavior of laboratory test specimens may not be pronounced and, therefore, the test results in terrestrial (1-g) environment may be sufficiently conclusive. However at low interparticle stresses, which can result either from low applied (confining) stresses or from excess pore fluid pressures developed within the soil mass without corresponding changes in the applied stresses, the presence of gravitational body forces acting on solid particles and interstitial fluids exerts a pronounced influence on movement of individual particles or particle groups. Such motions, in turn, cause changes in soil fabric which results in significant changes in the interparticle friction forces contributing to the soil's strength and deformation characteristics.

Alshibli, Khalid A.↗

Lower body negative pressure, experiment M092

Evaluation of orthostatic intolerance has been achieved by the use of lower body negative pressure (LBNP). The LBNP technique, though independent of gravity, simulates its effect by exposing the legs and the lower abdomen to reduced ambient pressures. This LBNP experiment, conducted during the 56-day simulation of the Skylab environment, was designed to supply baseline information on cardiovascular responses to periodic orthostatic stress. Impaired orthostatic tolerance, manifested by the increased heart rate, diminished systolic and pulse pressure, and increased tendency to syncope in the upright position, or during LBNP, was not observed in this experiment.

Johnson, R. L.↗

Characteristics of enzymatic induction provoked by chlordane

The effects of various stresses, such as restraint and lowering or raising of environmental temperature, in mice pretreated with chlordane were investigated. (Chlordane is an inhibitor of protein synthesis.) It was found that restraint or exposure to a cold environment for three hours mobilized the chlordane stored in the adipose tissue of mice.

Denys, A.↗

Overview of Boundary Layer Transition Research in Support of Orbiter Return To Flight

A predictive tool for estimating the onset of boundary layer transition resulting from damage to and/or repair of the thermal protection system was developed in support of Shuttle Return to Flight. The boundary layer transition tool is part of a suite of tools that analyze the aerothermodynamic environment to the local thermal protection system to allow informed disposition of damage for making recommendations to fly as is or to repair. Using mission specific trajectory information and details of each damage site or repair, the expected time (and thus Mach number) at transition onset is predicted to help define the aerothermodynamic environment to use in the subsequent thermal and stress analysis of the local thermal protection system and structure. The boundary layer transition criteria utilized for the tool was developed from ground-based measurements to account for the effect of both protuberances and cavities and has been calibrated against select flight data. Computed local boundary layer edge conditions were used to correlate the results, specifically the momentum thickness Reynolds number over the edge Mach number and the boundary layer thickness. For the initial Return to Flight mission, STS-114, empirical curve coefficients of 27, 100, and 900 were selected to predict transition onset for protuberances based on height, and cavities based on depth and length, respectively.

Berry, Scott A.↗

Creep and Environmental Durability of EBC/CMCs Under Imposed Thermal Gradient Conditions

Interest in SiC fiber-reinforced SiC ceramic matrix composite (CMC) environmental barrier coating (EBC) systems for use in high temperature structural applications has prompted the need for characterization of material strength and creep performance under complex aerospace turbine engine environments. Stress-rupture tests have been performed on SiC/SiC composites systems, with varying fiber types and coating schemes to demonstrate material behavior under isothermal conditions. Further testing was conducted under exposure to thermal stress gradients to determine the effect on creep resistance and material durability. In order to understand the associated damage mechanisms, emphasis is placed on experimental techniques as well as implementation of non-destructive evaluation; including electrical resistivity monitoring. The influence of environmental and loading conditions on life-limiting material properties is shown.

Appleby, Matthew↗

Veggie on Ice: The Effects of Plant Production on Human Behavioral Health Long-Duration Antarctic Overwintering Missions

Exploration class missions pose an increased risk of adverse crew behavioral health and psychiatric conditions. Identifying and developing effective and efficient countermeasure strategies are necessary to maintain crew health and performance. Previous studies (VEG-04A and VEG-04B) in the Veggie vegetable-production system on the International Space Station have evaluated the behavioral health effects of caring for plants as a potential countermeasure for crew stresses in spaceflight. The presence of plants can augment sensory stimulation (visual, tactile, and olfactory), provide a psychological link to Earth, reduce stress, and increase happiness. However, further research is needed to understand this strategy for longer-duration missions in isolated, confined, extreme (ICE) environments. The Neumayer III Station in Antarctica is one such environment that hosts a small crew for fourteen-month missions, of which eight to nine months are the remote overwintering phase. Neumayer III also features EDEN ISS, a greenhouse facility with 12.5 m2 plant cultivation space that, like Veggie, currently supplements the crew diet with a variety of fresh produce. Up to nine crew members of the 2021 overwintering mission will complete behavioral health surveys that are designed to capture enjoyment and time spent conducting various plant-related activities, sensory stimulation, and active and passive psychological effects of having live plants and available fresh food in an ICE environment. Data will be collected once pre-mission, up to ten times in-mission at monthly increments, and once post-mission. This study aims to quantify the impact of plant care as a countermeasure strategy for human spaceflight and Antarctic exploration missions.

J M Bunchek↗

The rhesus (Macaca mulatta) and crab-eating (Macaca fascicularis) monkeys in cardiovascular and aerospace research

Two nonhuman primate species were used to investigate the effects of gravitoinertial forces on pilot incapacitation and performance impairment, to define human physiologic tolerance and safe exposure limits to these environments, and to obtain data which can be used to evolve new methods to improve man's G tolerance to match the structural capability of new generation aircraft. The macaca fascicularis was used to study the effects of environmental stress and atheroscelerosis on cerebral blood flow and function agents on myocardial and cardiovascular function were studied in the macaca mulatta.

Erickson, H. H.↗