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At least 397 records · Page 22

Possible prebiotic significance of polyamines in the condensation, protection, encapsulation, and biological properties of DNA

Some properties of DNA condensed with spermidine have been compared with the properties of DNA condensed with Co3+(NH3)6 to determine whether condensation of DNA with these trivalent cations protects DNA against the action of DNase I and increases transcription and encapsulation of DNA into liposomes. It was shown that DNA condensed with Co3+(NH3)6 was resistant to the action of the endonuclease DNase I such as DNA condensed with spermidine was. However, DNA condensed with Co3+(NH3)6 was significantly less active in transcription with the E. coli RNA polymerase than DNA-spermidine condensed forms. In addition, it was demonstrated that both compacted forms of DNA were more efficiently encapsulated into neutral liposomes; however, negatively, charged liposomes were scarcely formed in the presence of DNA condensed with Co3+(NH3)6. These experiments and the well documented properties of polyamines increasing the resistance to radiations and hydrolysis of nucleic acids, as well as their biological activities, such as replication, transcription, and translation, together with the low concentration of Co3+ in the environment, lead us to propose spermidine as a plausible prebiotic DNA condensing agent rather than Co3+ and the basic proteins proposed by other authors. Then, we consider the possible role and relevance of the polyamine-nucleic acids complexes in the evolution of life.

NASA Discipline Exobiology↗

Evaluation of the Paratrend Multi-Analyte Sensor for Potential Utilization in Long-Duration Automated Cell Culture Monitoring

BACKGROUND: Compact and automated sensors are desired for assessing the health of cell cultures in biotechnology experiments. While several single-analyte sensors exist to measure culture health, a multi-analyte sensor would simplify the cell culture system. One such multi-analyte sensor, the Paratrend 7 manufactured by Diametrics Medical, consists of three optical fibers for measuring pH, dissolved carbon dioxide (pCO(2)), dissolved oxygen (pO(2)), and a thermocouple to measure temperature. The sensor bundle was designed for intra-vascular measurements in clinical settings, and can be used in bioreactors operated both on the ground and in NASA's Space Shuttle and International Space Station (ISS) experiments. METHODS: A Paratrend 7 sensor was placed at the outlet of a bioreactor inoculated with BHK-21 (baby hamster kidney) cells. The pH, pCO(2), pO(2), and temperature data were transferred continuously to an external computer. Cell culture medium, manually extracted from the bioreactor through a sampling port, was also assayed using a bench top blood gas analyzer (BGA). RESULTS: Two Paratrend 7 sensors were used over a single cell culture experiment (64 days). When compared to the manually obtained BGA samples, the sensor had good agreement for pH, pCO(2), and pO(2) with bias (and precision) 0.005(0.024), 8.0 mmHg (4.4 mmHg), and 11 mmHg (17 mmHg), respectively for the first two sensors. A third Paratrend sensor (operated for 141 days) had similar agreement (0.02+/-0.15 for pH, -4+/-8 mm Hg for pCO(2), and 24+/-18 mmHg for pO(2)). CONCLUSION: The resulting biases and precisions are com- parable to Paratrend sensor clinical results. Although the pO(2) differences may be acceptable for clinically relevant measurement ranges, the O(2) sensor in this bundle may not be reliable enough for the ranges of pO(2) in these cell culture studies without periodic calibration.

Validation Studies↗

Radiation-induced bystander effect and adaptive response in mammalian cells

Two conflicting phenomena, bystander effect and adaptive response, are important in determining the biological responses at low doses of radiation and have the potential to impact the shape of the dose-response relationship. Using the Columbia University charged-particle microbeam and the highly sensitive AL cell mutagenic assay, we show here that non-irradiated cells acquire mutagenesis through direct contact with cells whose nuclei have been traversed with a single alpha particle each. Pretreatment of cells with a low dose of X-rays four hours before alpha particle irradiation significantly decreased this bystander mutagenic response. Results from the present study address some of the fundamental issues regarding both the actual target and radiation dose effect and can contribute to our current understanding in radiation risk assessment. c2004 COSPAR. Published by Elsevier Ltd. All rights reserved.

NASA Discipline Radiation Health↗

DNA fragmentation induced by Fe ions in human cells: shielding influence on spatially correlated damage

Outside the magnetic field of the Earth, high energy heavy ions constitute a relevant part of the biologically significant dose to astronauts during the very long travels through space. The typical pattern of energy deposition in the matter by heavy ions on the microscopic scale is believed to produce spatially correlated damage in the DNA which is critical for radiobiological effects. We have investigated the influence of a lucite shielding on the initial production of very small DNA fragments in human fibroblasts irradiated with 1 GeV/u iron (Fe) ions. We also used gamma rays as reference radiation. Our results show: (1) a lower effect per incident ion when the shielding is used; (2) an higher DNA Double Strand Breaks (DSB) induction by Fe ions than by gamma rays in the size range 1-23 kbp; (3) a non-random DNA DSB induction by Fe ions. c2004 COSPAR. Published by Elsevier Ltd. All rights reserved.

NASA Discipline Radiation Health↗

Transcriptional profile of a myotube starvation model of atrophy

Skeletal muscle wasting is a pervasive phenomenon that can result from a wide range of pathological conditions as well as from habitual muscular inactivity. The present work describes a cell-culture condition that induces significant atrophy in skeletal muscle C2C12 myotubes. The failure to replenish differentiation media in mature myotubes leads to rapid atrophy (53% in diameter), which is referred to here as starvation. Affymetrix microarrays were used to develop a transcriptional profile of control (fed) vs. atrophied (nonfed) myotubes. Myotube starvation was characterized by an upregulation of genes involved in translational inhibition, amino acid biosynthesis and transport, and cell cycle arrest/apoptosis, among others. Downregulated genes included several structural and regulatory elements of the extracellular matrix as well as several elements of Wnt/frizzled and TGF-beta signaling pathways. Interestingly, the characteristic transcriptional upregulation of the ubiquitin-proteasome system, calpains, and cathepsins known to occur in multiple in vivo models of atrophy were not seen during myotube starvation. With the exception of the downregulation of extracellular matrix genes, serine protease inhibitor genes, and the upregulation of the translation initiation factor PHAS-I, this model of atrophy in cell culture has a transcriptional profile quite distinct from any study published to date with atrophy in whole muscle. These data show that, although the gross morphology of atrophied muscle fibers may be similar in whole muscle vs. myotube culture, the processes by which this phenotype is achieved differ markedly.

NASA Discipline Musculoskeletal↗

20V, 40 Ah Lithium Ion Polymer Battery for the Spacesuit

Objective: Consider a new battery design for EMU. Results: a) Electrovaya s aerospace cell production line is improving, but must further improve to achieve acceptable reliability; b) Completed functional, vibration, and thermal cycling of LIB; c) So far, electrical safety tests have produced good results; d) Completed functional, vibration, thermal cycling, power quality and EMI of LIB Charger; e) Completed CDR on 9/23/04; and f) Manufacturing Readiness Review for flight cell/battery production scheduled for Dec 04.

Darcy, Eric↗

Reducing Extra-Terrestrial Excavation Forces with Percussion

High launch costs and mission requirements drive the need for low mass excavators with mobility platforms, which in turn have little traction and excavation reaction capacity in low gravity environments. This presents the need for precursor and long term future missions with low mass robotic mining technology to perform In-Situ Resource Utilization (ISRU) tasks. This paper discusses a series of experiments that investigate the effectiveness of a percussive digging device to reduce excavation loads and thereby the mass of the excavator itself. The goal of percussive excavation is to fluidize dry regolith in front of the leading edge of the tool by mechanically separating the microscopic interlocking grains resulting in a reduced force needed to shear the soil. There are several variables involved with this technique; this experiment varied: Impact energy, frequency, and excavation speed and held constant: impact direction, depth of cut, angle of tool, and soil bulk density. The test apparatus consisted of an aluminum truss bridge with a central pivoting arm. Attached to the arm was a winch with a load cell in line that recorded the tension in the cable and therefore the excavation load. The arm could be adjusted for excavation depth which was recorded along with the arm angle relative to the bridge. A percussive mechanism and 30" wide pivoting bucket were attached at the end of the arm simulating a basic backhoe with a percussion direction tangent to the direction of . movement. Internally the mechanism used a set of die springs and barrel cam to produce the percussive blow. By changing the springs and the speed of the motor the impact energy and frequency of percussion could be varied independently. Impact energies from 11.2J to 30.5J and frequencies from 0 BPM to 700 BPM were investigated. A reduction in excavation force of as much as 51% was achieved in this experimental investigation. Smaller percussive digging implements, tested by others, have achieved a reduction of as much as 72%. This paper will examine the effects of impact energy, frequency, scaling and their effect on excavation forces in a dry granular material such as lunar regolith. The past several years have shown an increasing interest in mining space resources both for exploration and commercial enterprises. This work studied the benefits and risks of percussive excavation and prelimin~ry results indicate that this technique may become an enabling technology for extra-terrestrial excavation of regolith and ice.

Schuler, Jason↗

MHD Simulations of the Plasma Flow in the Magnetic Nozzle

The magnetohydrodynamic (MHD) flow of plasma through a magnetic nozzle is simulated by solving the governing equations for the plasma flow in the presence of an static magnetic field representing the applied nozzle. This work will numerically investigate the flow and behavior of the plasma as the inlet plasma conditions and magnetic nozzle field strength are varied. The MHD simulations are useful for addressing issues such as plasma detachment and to can be used to gain insight into the physical processes present in plasma flows found in thrusters that use magnetic nozzles. In the model, the MHD equations for a plasma, with separate temperatures calculated for the electrons and ions, are integrated over a finite cell volume with flux through each face computed for each of the conserved variables (mass, momentum, magnetic flux, energy) [1]. Stokes theorem is used to convert the area integrals over the faces of each cell into line integrals around the boundaries of each face. The state of the plasma is described using models of the ionization level, ratio of specific heats, thermal conductivity, and plasma resistivity. Anisotropies in current conduction due to Hall effect are included, and the system is closed using a real-gas equation of state to describe the relationship between the plasma density, temperature, and pressure.A separate magnetostatic solver is used to calculate the applied magnetic field, which is assumed constant for these calculations. The total magnetic field is obtained through superposition of the solution for the applied magnetic field and the self-consistently computed induced magnetic fields that arise as the flowing plasma reacts to the presence of the applied field. A solution for the applied magnetic field is represented in Fig. 1 (from Ref. [2]), exhibiting the classic converging-diverging field pattern. Previous research was able to demonstrate effects such as back-emf at a super-Alfvenic flow, which significantly alters the shape of the magnetic field in both the near- and far-field regions. However, in that work the downstream domain was constrained to a channel of constant cross-sectional area. In the present work we seek to address this issue by modeling the downstream region with a domain that permits free expansion of the plasma, permitting a better evaluation of the downstream effects the applied field has on the plasma. The inlet boundary conditions and applied magnetic field values will also be varied to determine the effect the initial plasma energy content and applied magnetic field energy density have on the near- and far-field plasma properties on the MHD code. This will determine the effect of inlet boundary conditions on the results downstream and address issues related to the restrictive numerical domain previously used.

Smith, T. E. R.↗

Supersonic Disk Gap Band Parachute Performance in the Wake of a Viking-Type Aeroshell from Mach 2 to 2.5

Supersonic wind tunnel testing of 0.813 m diameter Disk-Gap-Band parachutes is being conducted in the NASA Glenn Research Center (GRC) 10' x 10' wind-tunnel. The tests are conducted in support of the Mars Science Laboratory Parachute Decelerator System development and qualification. Four percent of full-scale parachutes were constructed similarly to the flight-article in material and construction techniques. The parachutes are attached to a 4% scale MSL entry-vehicle to simulate the free-flight configuration. The parachutes are tested from Mach 2 to 2.5 over a Reynolds number (Re) range of 1 to 3 x 10(exp 6), representative of the MSL deployment envelope. Constrained and unconstrained test configurations are investigated to quantify the effects of parachute trim, suspension line interaction, and alignment with the capsule wake. The parachute is constrained horizontally through the vent region, to measure canopy breathing and wake interaction for fixed trim angles of 0 and 10 degrees from the velocity vector. In the unconstrained configuration the parachute is permitted to trim and cone, similar to the free-flight varying its alignment relative to the entry-vehicle wake. Test diagnostics were chosen to quantify parachute performance and to provide insight into the flow field structure. An in-line load cell provided measurement of unsteady and mean drag as a function of Mach and Re. High-speed shadowgraph video of the upstream parachute flow field was used to capture bow-shock motion and stand of distance. Particle image velocimetry of the upstream parachute flow field provides spatially and temporally resolved measurement velocity and turbulent statistics. Multiple high speed video views of targets placed in the interior of the canopy enable photo-grammetric measurement of the fabric motion in time and space from reflective. High speed video is also used to document the supersonic inflation and measure trim angle, projected area, and frequency of area oscillations.

EDL↗

Photogrammetric Deflection Measurements for the Tiltrotor Test Rig (TTR) Multi-Component Rotor Balance Calibration

Calibrating the internal, multi-component balance mounted in the Tiltrotor Test Rig (TTR) required photogrammetric measurements to determine the location and orientation of forces applied to the balance. The TTR, with the balance and calibration hardware attached, was mounted in a custom calibration stand. Calibration loads were applied using eleven hydraulic actuators, operating in tension only, that were attached to the forward frame of the calibration stand and the TTR calibration hardware via linkages with in-line load cells. Before the linkages were installed, photogrammetry was used to determine the location of the linkage attachment points on the forward frame and on the TTR calibration hardware. Photogrammetric measurements were used to determine the displacement of the linkage attachment points on the TTR due to deflection of the hardware under applied loads. These measurements represent the first photogrammetric deflection measurements to be made to support 6-component rotor balance calibration. This paper describes the design of the TTR and the calibration hardware, and presents the development, set-up and use of the photogrammetry system, along with some selected measurement results.

Calibration↗

Photogrammetric Deflection Measurements for the Tiltrotor Test Rig (TTR) Multi-Component Rotor Balance Calibration

Calibrating the internal, multi-component balance mounted in the Tiltrotor Test Rig (TTR) required photogrammetric measurements to determine the location and orientation of forces applied to the balance. The TTR, with the balance and calibration hardware attached, was mounted in a custom calibration stand. Calibration loads were applied using eleven hydraulic actuators, operating in tension only, that were attached to the forward frame of the calibration stand and the TTR calibration hardware via linkages with in-line load cells. Before the linkages were installed, photogrammetry was used to determine the location of the linkage attachment points on the forward frame and on the TTR calibration hardware. Photogrammetric measurements were used to determine the displacement of the linkage attachment points on the TTR due to deflection of the hardware under applied loads. These measurements represent the first photogrammetric deflection measurements to be made to support 6-component rotor balance calibration. This paper describes the design of the TTR and the calibration hardware, and presents the development, set-up and use of the photogrammetry system, along with some selected measurement results.

Calibration↗

Benchmarking Computational Tools for Calling SNPs and Indels in Complex Microbial Populations

The NASA BioNutrients missions seek to understand the suitability of microorganisms for bioproduction during space flight. One topic of interest is the stability of microbial genomes during long-term ambient storage and subsequent rehydration and growth. To address these questions, samples from 8 species were flown to ISS for 5 years of desiccated storage at ambient temperature (Stasis Packs) and 2 species were packaged along with powdered media inside a bioreactor system to allow hydration and growth in microgravity (Production Packs). For both systems, Whole Genome Sequencing (WGS) of the DNA extracted from the returned samples and paired ground controls will be conducted to identify changes in genome stability due to time, storage conditions and growth in space. Across the technical replicates, ground controls, 10 timepoints, and multiple experimental conditions, ~300 samples have been selected for initial analysis with WGS sequencing to 100x coverage. A flexible and resource efficient mutation calling pipeline is needed to process this large dataset and allow for comparisons between species. Many bioinformatics tools for calling Indels and Single Nucleotide Variants (SNVs) are designed for use with pure isolates, where true variations from the reference genome are expected to dominate the reads aligning to the location of mutation. In contrast, DNA from the Stasis Pack (SP) samples was collected directly after recovery from desiccated storage and the Production Pack (PP) samples were collected after fermentation. In this context, reads with mutations are expected to be less frequent than reads that align with the reference genome, as each sample will include multiple lines of cells. Thus, BioNutrients samples are expected to be similar to samples from cancer cell or “pooled” sequencing approaches. In preparation for the analysis of the BioNutrients samples, we have tested three mutation calling tools (GATK for Microbes, BreSeq and DiscoSNP) designed for complex samples. A challenge of validating mutation identification pipelines is a lack of “Ground Truth” datasets, especially for complex samples. To compare these three tools, we sought to identify mutations in pre-existing WGS data collected from populations of Chlamydomonas reinhardtii that were exposed to UV mutagenesis and growth in LEO as part of the Space Algae-1 mission. Here we present a summary of these tools against the analysis originally conducted using the CRISP tool. Critical metrics are compared such as runtime, the number of SNPs, the number and size of Indels, and patterns of transversion and transitions identified by each tool are reported. By sharing these benchmarking results collected in support of the BioNutrients mission, we aim to guide others seeking to identify SNVs in similarly complex microbial samples.

Biology↗

Homologous recombination as a potential target for caffeine radiosensitization in mammalian cells: reduced caffeine radiosensitization in XRCC2 and XRCC3 mutants

The radiosensitizing effect of caffeine has been associated with the disruption of multiple DNA damage-responsive cell cycle checkpoints, but several lines of evidence also implicate inhibition of DNA repair. The role of DNA repair inhibition in caffeine radiosensitization remains uncharacterized, and it is unknown which repair process, or lesion, is affected. We show that a radiosensitive cell line, mutant for the RAD51 homolog XRCC2 and defective in homologous recombination repair (HRR), displays significantly diminished caffeine radiosensitization that can be restored by expression of XRCC2. Despite the reduced radiosensitization, caffeine effectively abrogates checkpoints in S and G2 phases in XRCC2 mutant cells indicating that checkpoint abrogation is not sufficient for radiosensitization. Another radiosensitive line, mutant for XRCC3 and defective in HRR, similarly shows reduced caffeine radiosensitization. On the other hand, a radiosensitive mutant (irs-20) of DNA-PKcs with a defect in non-homologous end-joining (NHEJ) is radiosensitized by caffeine to an extent comparable to wild-type cells. In addition, rejoining of radiation-induced DNA DSBs, that mainly reflects NHEJ, remains unaffected by caffeine in XRCC2 and XRCC3 mutants, or their wild-type counterparts. These observations suggest that caffeine targets steps in HRR but not in NHEJ and that abrogation of checkpoint response is not sufficient to explain radiosensitization. Indeed, immortalized fibroblasts from AT patients show caffeine radiosensitization despite the checkpoint defects associated with ATM mutation. We propose that caffeine radiosensitization is mediated by inhibition of stages in DNA DSB repair requiring HRR and that checkpoint disruption contributes by allowing these DSBs to transit into irreparable states. Thus, checkpoints may contribute to genomic stability by promoting error-free HRR.

NASA Discipline Radiation Health↗

Flexibility in space solar cell production

The wide range of cells that must be available from present-day production lines for space solar cells are described. After over thirty years of space-cell use, there is very little standardization in solar cell design. It is not generally recognized what a wide range of designs must remain available on cell production lines. This range of designs is surveyed.

Khemthong, Scott↗

Remote sensing of stratospheric and mesospheric winds by gas correlation electrooptic phase-modulation spectroscopy

A method is developed for directly measuring atmospheric winds in the 20-120 km altitude interval from a spacecraft. The wind sensor measures the Doppler shift between the spectral absorption lines of a gas in a cell within the instrument and the thermal emission lines of the same gas in the atmosphere. The wind measurements are performed using a spacecraft-borne gas correlation spectrometer viewing the limb of the atmosphere. The wind-induced Doppler shift between the two spectra, and thus the magnitude of the wind itself, is obtained by phase modulating the incoming thermal radiation (equivalent to frequency modulation) by means of an electrooptically active crystal to determine the frequency shift required to reestablish exact correlation between the lines in the cell and the lines from the atmosphere. Results of numerical simulations of the wind-sensor performance indicate that the noise-equivalent-wind is between 1-5 m/s over most of the stratosphere and mesosphere.

Mccleese, D. J.↗

Raman-shifted dye laser for water vapor DIAL measurements

For improved DIAL measurements of water vapor in the upper troposphere or lower stratosphere, narrowband (about 0.03/cm) laser radiation at 720- and 940-nm wavelengths was generated by stimulated Raman scattering (SRS), using the narrow linewidth (about 0.02/cm) output of a Nd:YAG-pumped dye laser. For a hydrogen pressure of 350 psi, the first Stokes conversion efficiencies to 940 nm were 20 percent and 35 percent, when using a conventional and waveguide Raman cell, respectively. The linewidth of the first Stokes line at high cell pressures, and the inferred collisional broadening coefficients, agree well with those previously measured in spontaneous Raman scattering.

Grossmann, B. E.↗