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Binding of Campylobacter jejuni FliW Adjacent to the CsrA RNA-Binding Pockets Modulates CsrA Regulatory Activity

Campylobacter jejuni CsrA is an mRNA-binding, post-transcriptional regulator that controls many metabolic- and virulence-related characteristics of this important pathogen. In contrast to E. coli CsrA, whose activity is modulated by binding to small non-coding RNAs (sRNAs), C. jejuni CsrA activity is controlled by binding to the CsrA antagonist FliW. In this study, we identified the FliW binding site on CsrA. Deletion of the C-terminus of C. jejuni CsrA, which is extended relative to sRNA-binding CsrA proteins, abrogated FliW binding. Bacterial two-hybrid experiments were used to assess the interaction of FliW with wild-type CsrA and mutants thereof, in which every amino acid was individually mutated. Two CsrA mutations (V51A and N55A) resulted in a significant decrease in FliW binding. The V51A and N55A mutants also showed a decrease in CsrA-FliW complex formation, as assessed by size-exclusion chromatography and surface plasmon resonance. These residues were highly conserved in bacterial species containing CsrA orthologs whose activities are predicted to be regulated by FliW. The location of FliW binding was immediately adjacent to the two RNA-binding sites of the CsrA homodimer, suggesting the model that FliW binding to CsrA modulates its ability to bind to its mRNA targets either by steric hindrance, electrostatic repulsion, or by altering the overall structure of the RNA-binding sites.

59 BASIC BIOLOGICAL SCIENCES↗

Integrated Locomotor Function Tests for Countermeasure Evaluation

Following spaceflight crewmembers experience locomotor dysfunction due to inflight adaptive alterations in sensorimotor function. Countermeasures designed to mitigate these postflight gait alterations need to be assessed with a new generation of tests that evaluate the interaction of various sensorimotor sub-systems central to locomotor control. The goal of the present study was to develop new functional tests of locomotor control that could be used to test the efficacy of countermeasures. These tests were designed to simultaneously examine the function of multiple sensorimotor systems underlying the control of locomotion and be operationally relevant to the astronaut population. Traditionally, gaze stabilization has been studied almost exclusively in seated subjects performing target acquisition tasks requiring only the involvement of coordinated eye-head movements. However, activities like walking involve full-body movement and require coordination between lower limbs and the eye-head-trunk complex to achieve stabilized gaze during locomotion. Therefore the first goal of this study was to determine how the multiple, interdependent, full-body sensorimotor gaze stabilization subsystems are functionally coordinated during locomotion. In an earlier study we investigated how alteration in gaze tasking changes full-body locomotor control strategies. Subjects walked on a treadmill and either focused on a central point target or read numeral characters. We measured: temporal parameters of gait, full body sagittal plane segmental kinematics of the head, trunk, thigh, shank and foot, accelerations along the vertical axis at the head and the shank, and the vertical forces acting on the support surface. In comparison to the point target fixation condition, the results of the number reading task showed that compensatory head pitch movements increased, peak head acceleration was reduced and knee flexion at heel-strike was increased. In a more recent study we investigated the adaptive remodeling of the full-body gaze control systems following exposure to visual-vestibular conflict. Subjects walked on a treadmill before and after a 30- minute exposure to 0.5X minifying during which self-generated sinusoidal vertical head rotations were performed while seated. Following exposure to visual-vestibular conflict subjects showed a restriction in compensatory head movements, increased knee and ankle flexion after heel-strike and a decrease in the rate of body loading during the rapid weight transfer phase after the heel strike event. Taken together, results from both studies provide evidence that the full body contributes to gaze stabilization during locomotion, and that different functional elements are responsive to changes in visual task constraints and are subject to adaptive alterations following exposure to visual-vestibular conflict. This information provides the basis for the design of a new generation of integrative tests that incorporate the evaluation of multiple neural control systems relevant to astronaut operational performance.

Bloomberg, J. J.↗

International Space Station (ISS) Risk Reduction Activities

As the assembly of the ISS nears completion, it is worthwhile to step back and review some of the actions pursued by the Program in recent years to reduce risk and enhance the safety and health of ISS crewmembers, visitors, and space flight participants. While the ISS requirements and initial design were intended to provide the best practicable levels of safety, it is always possible to reduce risk -- given the determination and commitment to do so. The following is a summary of some of the steps taken by the ISS Program Manager, by our International Partners, by hardware and software designers, by operational specialists, and by safety personnel to continuously enhance the safety of the ISS. While decades of work went into developing the ISS requirements, there are many things in a Program like the ISS that can only be learned through actual operational experience. These risk reduction activities can be divided into roughly three categories: (1) Areas that were initially noncompliant which have subsequently been brought into compliance or near compliance (i.e., Micrometeoroid and Orbital Debris [MMOD] protection, acoustics) (2) Areas where initial design requirements were eventually considered inadequate and were subsequently augmented (i.e., Toxicity Level 4 materials, emergency hardware and procedures) (3) Areas where risks were initially underestimated, and have subsequently been addressed through additional mitigation (i.e., Extravehicular Activity [EVA] sharp edges, plasma shock hazards) Due to the hard work and cooperation of many parties working together across the span of nearly a decade, the ISS is now a safer and healthier environment for our crew, in many cases exceeding the risk reduction targets inherent in the intent of the original design. It will provide a safe and stable platform for utilization and discovery.

Fodroci, Michael↗

NASA’s Earth System Observatory Formulation Progress

The 2017 Earth Science Decadal Survey by the National Academies of Science, Engineering, and Medicine identified five foundational observations, or Designated Observables (DO), to be implemented as missions by NASA. To address these five DO areas – Surface Biology and Geology; Mass Change; Aerosols; Cloud, Convection, and Precipitation; and Surface Deformation and Change – NASA is developing the Earth System Observatory (ESO) as an array of Earth-focused, interconnected satellite missions. The ESO will build on the strong history of international partnerships in NASA Earth science, with initial participation and collaboration on these missions across space agency partners including the Japanese Aerospace Exploration Agency (JAXA), Centre National D'Etudes Spatiales (CNES), Canadian Space Agency (CSA), Deutsches Zentrum für Luft- und Raumfahrt (DLR), and Agenzia Spaziale Italiana (ASI). Targeting launch dates in the late 2020s and early 2030s, the ESO will work as a single observatory, with each satellite in the ESO delivering its own valuable information, but taken together, the data and imagery will provide the global community with a multidimensional, holistic view of the Earth. In addition to furthering the world’s scientific knowledge, data gathered by ESO missions will provide actionable science to inform decisions related to climate change, disaster mitigation, and wildfires, improve real-time agricultural processes, and many other applications. This paper will provide an overview of the ESO development activities, as well as an update on the individual ESO missions currently in formulation – Atmosphere Observing System, Gravity Recovery and Climate Experiment-Continuity, and Surface Biology and Geology – including current mission architectures and concepts, international partner collaboration, science community engagement, and applications efforts, as well as how ESO data will be freely open and accessible to users all over the world.

NASA Earth Science↗

Polymerization and editing modes of a high-fidelity DNA polymerase are linked by a well-defined path

Proofreading by replicative DNA polymerases is a fundamental mechanism ensuring DNA replication fidelity. In proofreading, mis-incorporated nucleotides are excised through the 3'-5' exonuclease activity of the DNA polymerase holoenzyme. The exonuclease site is distal from the polymerization site, imposing stringent structural and kinetic requirements for efficient primer strand transfer. Yet, the molecular mechanism of this transfer is not known. Here we employ molecular simulations using recent cryo-EM structures and biochemical analyses to delineate an optimal free energy path connecting the polymerization and exonuclease states of E. coli replicative DNA polymerase Pol III. We identify structures for all intermediates, in which the transitioning primer strand is stabilized by conserved Pol III residues along the fingers, thumb and exonuclease domains. We demonstrate switching kinetics on a tens of milliseconds timescale and unveil a complete pol-to-exo switching mechanism, validated by targeted mutational experiments.

59 BASIC BIOLOGICAL SCIENCES↗

The Sun in Time

Using a grant from NASA's Initiative to Develop Education through Astronomy and Space Science (IDEAS) program, we have developed an inter-disciplinary curriculum for middle-school students which targets both history and astronomy. Our curriculum explores the attitudes and techniques of ancient spiritual leaders, specifically those of the Maya and Inca cultures, who observed and tried to control the Sun. We wish students to understand the probable importance of astronomical observations to these ancient peoples. In addition, using the experience of an archaeologist, we show how modern techniques of viewing the Earth through satellite imagery, has allowed the re-discovery of ancient sites where solar observations and attempted manipulation of the universe took place. To contrast ancient observations of the Sun with modern ones, we use the experience of a solar astronomer and bring to the classroom up-to-date information about solar astronomy and the impact of solar activity on the Earth's environment. In this presentation, we will present fragments of our curriculum as well as results from pre- and post-tests given to participating groups of students. Finally, we will discuss comments from local middle-school teachers who were asked to evaluate our curriculum.

Adams, Mitzi L.↗

SONEX Airborne Mission and Coordinated POLINAT-2 Activity: Overview and Accomplishments

The SASS (Subsonic Assessment) Ozone and NO(x) Experiment (SONEX) was an airborne field campaign conducted in October-November 1997 in the vicinity of the North Atlantic Flight Corridor Lo study the impact of aircraft emissions on NOx and ozone (03). A fully instrumented NASA DC-8 aircraft was used as the primary SONEX platform. SONEX activities were closely coordinated with the European POLINAT-2 (Pollution from Aircraft Emissions in the North Atlantic Flight Corridor) program, which used a Falcon-20 aircraft and an instrumented in-service Swissair B-747. Both campaigns focused on the upper troposphere/"lowermost" stratosphere (UT/LS) as the region of greatest interest. Specific sampling goals were achieved with the aid of a state-of-the art modeling and meteorological support system, which allowed targeted sampling of air parcels with desired characteristics. A substantial impact of aircraft emissions on NO(x) and O3 in the UT/LS of the study region is shown to be present. It is further shown that the NO(x)- HO(x)-O3 relationships are highly nonlinear and must be accurately simulated to make meaningful future predictions with global models. SONEXIPOLINAT-2 results are being published in Special Sections of GRL and JGR. Here we provide a brief overview of SONEX design, implementation, and expected results to provide a context within which these publications can be understood.

Singh, Hanwant B.↗

NASA's Lunar Atmosphere and Dust Environment Explorer (LADEE)

Nearly 40 years have passed since the last Apollo missions investigated the mysteries of the lunar atmosphere and the question of levitated lunar dust. The most important questions remain: what is the composition, structure and variability of the tenuous lunar exosphere? What are its origins, transport mechanisms, and loss processes? Is lofted lunar dust the cause of the horizon glow observed by the Surveyor missions and Apollo astronauts? How does such levitated dust arise and move, what is its density, and what is its ultimate fate? The US National Academy of Sciences/National Research Council decadal surveys and the recent "Scientific Context for Exploration of the Moon" (SCEM) reports have identified studies of the pristine state of the lunar atmosphere and dust environment as among the leading priorities for future lunar science missions. These measurements have become particularly important since recent observations by the Lunar Crater Observation and Sensing Satellite (LCROSS) mission point to significant amounts of water and other volatiles sequestered within polar lunar cold traps. Moreover Chandrayaan/M3, EPOXI and Cassini/VIMS have identified molecular water and hydroxyl on lunar surface regolith grains. Variability in concentration suggests these species are likely to be present in the exosphere, and thus constitute a source for the cold traps. NASA s Lunar Atmosphere and Dust Environment Explorer (LADEE) is currently under development to address these goals. LADEE will determine the composition of the lunar atmosphere and investigate the processes that control its distribution and variability, including sources, sinks, and surface interactions. LADEE will also determine whether dust is present in the lunar exosphere, and reveal its sources and variability. LADEE s results are relevant to surface boundary exospheres and dust processes throughout the solar system, will address questions regarding the origin and evolution of lunar volatiles, and will have implications for future exploration activities. LADEE will be the first mission based on the Ames Common Bus design. LADEE employs a high heritage instrument payload: a Neutral Mass Spectrometer (NMS), an Ultraviolet/Visible Spectrometer (UVS), and the Lunar Dust Experiment (LDEX). It will also carry a space terminal as part of the Lunar Laser Communication Demonstration (LLCD), which is a technology demonstration. LLCD will also supply a ground terminal. LLCD is funded by the Space Operations Mission Directorate (SOMD), managed by GSFC, and built by MIT Lincoln Lab. NMS was directed to the Goddard Space Flight Center (GSFC) and UVS to Ames Research Center (ARC). LDEX was selected through the Stand Alone Missions of Opportunity Notice (SALMON) Acquisition Process, and is provided by the University of Colorado at Boulder. The LADEE NMS covers a m/z range of 2-150 and draws its design from mass spectrometers developed at GSFC for the MSL/SAM, Cassini Orbiter, CONTOUR, and MAVEN missions. The UVS instrument is a next-generation, high-reliability version of the LCROSS UV-Vis spectrometer, spanning 250-800 nm wavelength, with high (<1 nm) spectral resolution. UVS will also perform dust occultation measurements via a solar viewer optic. LDEX senses dust impacts in situ, at LADEE orbital altitudes of 50 km and below, with a particle size range of between 100 nm and 5 micron. Dust particle impacts on a large hemispherical target create electron and ion pairs. The latter are focused and accelerated in an electric field and detected at a microchannel plate. LADEE is an important part of NASA s portfolio of near-term lunar missions; launch is planned for May, 2013. The lunar atmosphere is the most accessible example of a surface boundary exosphere, and may reveal the sources and cycling of volatiles. Dynamic dust activity must be accounted for in the design and operation of lunar surface operations.

Elphic, Richard↗

The ePix10k 2-megapixel hard X-ray detector at LCLS

The ePix10ka2M (ePix10k) is a new large area detector specifically developed for X-ray free-electron laser (XFEL) applications. The hybrid pixel detector was developed at SLAC to provide a hard X-ray area detector with a high dynamic range, running at the 120 Hz repetition rate of the Linac Coherent Light Source (LCLS). The ePix10k consists of 16 modules, each with 352 × 384 pixels of 100 µm × 100 µm distributed on four ASICs, resulting in a 2.16 megapixel detector, with a 16.5 cm × 16.5 cm active area and ∼80% coverage. The high dynamic range is achieved with three distinct gain settings (low, medium, high) as well as two auto-ranging modes (high-to-low and medium-to-low). Here the three fixed gain modes are evaluated. The resulting dynamic range (from single photon counting to 10000 photons pixel −1 pulse −1 at 8 keV) makes it suitable for a large number of different XFEL experiments. The ePix10k replaces the large CSPAD in operation since 2011. The dimensions of the two detectors are similar, making the upgrade from CSPAD to ePix10k straightforward for most setups, with the ePix10k improving on experimental performance. The SLAC-developed ePix cameras all utilize a similar platform, are tailored to target different experimental conditions and are designed to provide an upgrade path for future high-repetition-rate XFELs. Here the first measurements on this new ePix10k detector are presented and the performance under typical XFEL conditions evaluated during an LCLS X-ray diffuse scattering experiment measuring the 9.5 keV X-ray photons scattered from a thin liquid jet.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

IXPE: The Imaging X-ray Polarimetry Explorer, Implementing a Dedicated Polarimetry Mission

Only a few experiments have conducted x-ray polarimetry of cosmic sources since Weisskopf et al confirmed the 19% polarization of the Crab Nebula with the Orbiting Solar Observatory (OSO-8) in the 70's center dot The challenge is to measure a faint polarized component against a background of non-polarized signal (as well as the other, typical background components) center dot Typically, for a few % minimum detectable polarization, 106 photons are required. center dot So, a dedicated mission is vital with instruments that are designed specifically to measure polarization (with minimal systematic effects) Over the proposed mission life (2- 3 years), IXPE will first survey representative samples of several categories of targets: magnetars, isolated pulsars, pulsar wind nebula and supernova remnants, microquasars, active galaxies etc. The survey results will guide detailed follow-up observations. Precise calibration of IXPE is vital to ensuring sensitivity goals are met. The detectors will be characterized in Italy, and then a full calibration of the complete instrument will be performed at MSFC's stray light facility. Polarized flux at different energies Heritage: X-ray Optics at MSFC polarimetry mission.

Ramsey, Brian↗

Structure and Interactions of HIV-1 gp41 CHR-NHR Reverse Hairpin Constructs Reveal Molecular Determinants of Antiviral Activity

Engineered reverse hairpin constructs containing a partial C-heptad repeat (CHR) sequence followed by a short loop and full-length N-heptad repeat (NHR) were previously shown to form trimers in solution and to be nanomolar inhibitors of HIV-1 Env mediated fusion. Their target is the in situ gp41 fusion intermediate, and they have similar potency to other previously reported NHR trimers. However, their design implies that the NHR is partially covered by CHR, which would be expected to limit potency. An exposed hydrophobic pocket in the folded structure may be sufficient to confer the observed potency, or they may exist in a partially unfolded state exposing full length NHR. Here, in this study, we examined their structure by crystallography, CD and fluorescence, establishing that the proteins are folded hairpins both in crystal form and in solution. We examined unfolding in the milieu of the fusion reaction by conducting experiments in the presence of a membrane mimetic solvent and by engineering a disulfide bond into the structure to prevent partial unfolding. We further examined the role of the hydrophobic pocket, using a hairpin-small molecule adduct that occluded the pocket, as confirmed by X-ray footprinting. The results demonstrated that the NHR region nominally covered by CHR in the engineered constructs and the hydrophobic pocket region that is exposed by design were both essential for nanomolar potency and that interaction with membrane is likely to play a role in promoting the required inhibitor structure. The design concepts can be applied to other Class 1 viral fusion proteins.

59 BASIC BIOLOGICAL SCIENCES↗

Connecting Material Properties and Redox Flow Cell Cycling Performance through Zero-Dimensional Models

Improvements in redox flow battery (RFB) performance and durability can be achieved through the development of new active materials, electrolytes, and membranes. While a rich design space exists for emerging materials, complex tradeoffs challenge the articulation of unambiguous target criteria, as the relationships between component selection and cycling performance are multifaceted. Here, we derive zero-dimensional, analytical expressions for mass balances and cell voltages under galvanostatic cycling, enabling direct connections between material/electrolyte properties, cell operating conditions, and resulting performance metrics (e.g., energy efficiency, capacity fade). To demonstrate the utility of this modeling framework, we highlight several considerations for RFB design, including upper bound estimation, active species decay, and membrane/separator conductivity-selectivity tradeoffs. Furthermore, we also discuss modalities for extending this framework to incorporate kinetic losses, distributed ohmic losses, and multiple spatial domains. Importantly, because the mass balances are solved analytically, hundreds of cycles can be simulated in seconds, potentially facilitating detailed parametric sweeps, system optimization, and parameter estimation from cycling experiments. More broadly, this approach provides a means for assessing the impact of cell components that simultaneously influence multiple performance-defining processes, aiding in the elucidation of key descriptors and the identification of favorable materials combinations for specific applications.

25 ENERGY STORAGE↗

Gamma-ray and millimeter-wave emissions from the 1991 June X-class solar flares

We have studied the spectacular 1991 June X-class flares using gamma-ray data from the Charged Particle Detectors (CPDs) of the Burst and Transient Source Experiment (BATSE) on the Compton Gamma Ray Observatory (CGRO) and 80 GHz millimeter data from Nobeyama, Japan. The CPDs were the only CGRO instrument that did not saturate during the extremely intense 1991 June 4 flare. We have shown that for this flare the CPDs respond to MeV photons, most of which are due to bremsstrahlung produced by relativistic electrons at the Sun. We have further shown that the gamma-ray and millimeter observations agree numerically if the 80 GHz radiation is gyrosynchrotron radiation produced by trapped electrons and the gamma rays are thick-target bremsstrahlung due to electrons precipitating out of the trap. The requirement that the trapping time obtained from the numerical comparison be consistent with the observed time profiles implies a magnetic field between about 200 and 300 G and an electron spectral index between about 3 to 5. By comparing the CPD observations with both the 80 GHz data and nuclear line data from the Energetic Gamma Ray Experiment Telescope (EGRET) and the Oriented Scintillation Spectroscopy Experiment (OSSE) on CGRO for the flares of June 4, 6, 9, and 11, we found that the ratio of the CPD counts to both the millimeter flux densities and the nuclear line fluences decreases with decreasing flare heliocentric angle. All of these flares were produced in the same active region. We interpreted this result in terms of a loop model in which the gyrosynchrotron emission is produced in the coronal portion of the loop where the electrons are kept isotropic by pitch angle scattering due to plasma turbulence, while the bremsstrahlung is produced by precipitating electrons that interact anisotropically. We found that the trapping time in the coronal portion is time dependent, reaching a minimum of about 10 s at the peak of the CPD count rate. We suggested the damping of the turbulence as a possible reason for the variation of the trapping time. turbulence as a possible reason for the variation of the trapping time.

Ramaty, Reuven↗

Improved Spacecraft Materials for Radiation Protection

Methods by which radiation shielding is optimized need to be developed and materials of improved shielding characteristics identified and validated. The galactic cosmic rays (GCR) are very penetrating and the energy absorbed by the astronaut behind the shield is nearly independent of shield composition and even the shield thickness. However, the mix of particles in the transmitted beam changes rapidly with shield material composition and thickness. This results in part from the breakup of the high-energy heavy ions of the GCR which make contributions to biological effects out of proportion to their deposited energy. So the mixture of particles in the radiation field changes with shielding and the control of risk contributions from dominant particle types is critical to reducing the hazard to the astronaut. The risk of biological injury for a given particle type depends on the type of biological effect and is specific to cell or tissue type. Thus, one is faced with choosing materials which may protect a given tissue against a given effect but leave unchanged or even increase the risk of other effects in the same tissue or increase the risks to other adjacent tissues of a different type in the same individual. The optimization of shield composition will then be tied to a specific tissue and risk to that tissue. Such peculiarities arise from the complicated mixture of particles, the nature of their biological response, and the details of their interaction with material constituents. Aside from the understanding of the biological response to specific components, one also needs an accurate understanding of the radiation emerging from the shield material. This latter subject has been a principal element of this project. In the past ten years our understanding of space radiation interactions with materials has changed radically, with a large impact on shield design. For example, the NCRP estimated that only 2 g/sq cm. of aluminum would be required to meet the annual 500 mSv limit for the exposure of the blood forming organs (this limit is strictly for LEO but can be used as a guideline for the Mars mission analysis). The current estimates require aluminum shield thicknesses above 50 g/sq cm., which is impractical. In such a heavily shielded vehicle, the neutrons produced throughout the vehicle also contribute significantly to the exposure and this demands greater care in describing the angular dependence of secondary particle production processes. As such the continued testing of databases and transport procedures in laboratory and spaceflight experiments has continued. This has been the focus of much of the last year's activity and has resulted in improved neutron prediction capability. These new methods have also improved our understanding of the surface environment of Mars. The Mars 2003 NRA HEDS related surface science requirements were driven by the need to validate predictions on the upward flux of neutrons produced in the Martian regolith and bedrock made by the codes developed under this project. The codes used in the surface environment definition are also being used to look at in situ resources for the development of construction material for Martian surface facilities. For example, synthesis of polyimides and polyethylene as binders of regolith for developing basic structural elements has been studied and targets built for accelerator beam testing of radiation shielding properties. Preliminary mechanical tests have also been promising. Improved spacecraft materials have been identified (using the criteria reported by this project at the last conference) as potentially important for future shielding materials. These are liquid hydrogen, hydrogenated nanofibers, liquid methane, LiH, Polyethylene, Polysulfone, and Polyetherimide (in order of decreasing shield performance). Some of the materials are multifunctional and are required for other onboard systems. We are currently preparing software for trade studies with these materials relative to the Mars Reference Mission as required in the project's final year.

Wilson, John W.↗

Long-Term International Space Station (ISS) Risk Reduction Activities

As the assembly of the ISS nears completion, it is worthwhile to step back and review some of the actions pursued by the Program in recent years to reduce risk and enhance the safety and health of ISS crewmembers, visitors, and space flight participants. While the initial ISS requirements and design were intended to provide the best practicable levels of safety, it is always possible to further reduce risk given the determination, commitment, and resources to do so. The following is a summary of some of the steps taken by the ISS Program Manager, by our International Partners, by hardware and software designers, by operational specialists, and by safety personnel to continuously enhance the safety of the ISS, and to reduce risk to all crewmembers. While years of work went into the development of ISS requirements, there are many things associated with risk reduction in a Program like the ISS that can only be learned through actual operational experience. These risk reduction activities can be divided into roughly three categories: Areas that were initially noncompliant which have subsequently been brought into compliance or near compliance (i.e., Micrometeoroid and Orbital Debris [MMOD] protection, acoustics) Areas where initial design requirements were eventually considered inadequate and were subsequently augmented (i.e., Toxicity hazard level-4 materials, emergency procedures, emergency equipment, control of drag-throughs) Areas where risks were initially underestimated, and have subsequently been addressed through additional mitigation (i.e., Extravehicular Activity [EVA] sharp edges, plasma shock hazards). Due to the hard work and cooperation of many parties working together across the span of more than a decade, the ISS is now a safer and healthier environment for our crew, in many cases exceeding the risk reduction targets inherent in the intent of the original design. It will provide a safe and stable platform for utilization and discovery for years to come.

Forroci, Michael P.↗

Modeling and design of a separate effects irradiation test targeting fission gas release from Cr-doped UO 2

Fission gas release (FGR) from nuclear fuel during operation can diminish heat transfer properties across the pellet-cladding gap and increase the fuel rod internal pressure, thereby posing a concern to fuel reliability and safety during an accident. Enlarging the fuel grain size, which has been shown to improve fission gas retention, can be achieved by doping the fuel feedstock prior to sintering. In this work, the BISON fuel performance code was used to predict FGR from undoped and chromia-doped UO 2 (referred to as Cr-doped UO 2 ) fuel specimens with different grain sizes and across various temperatures. The BISON models identified the irradiation conditions for which FGR is most significant, and a separate effects irradiation experiment in the High Flux Isotope Reactor (HFIR) was then developed targeting those conditions. Further, the experiment leveraged the MiniFuel irradiation capability at Oak Ridge National Laboratory and consisted of 12 fuel specimens of varying grain size and Cr content. A coupling scheme between BISON FGR results and the ANSYS finite element thermal model used for experiment design was formulated to predict cumulative FGR from each fuel specimen based on expected irradiation temperature histories. The fuel samples were fabricated and characterized as a part of this work, and the fuel compositions modeled in BISON were representative of the specimens used in the experiment. This combined modeling and experimental effort aims to study the effect of fuel grain size and Cr content on FGR and to provide simulated BISON FGR results that can be used for future model validation activities.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

Targeted mutagenesis and high-throughput screening of diversified gene and promoter libraries for isolating gain-of-function mutations

Targeted mutagenesis of a promoter or gene is essential for attaining new functions in microbial and protein engineering efforts. In the burgeoning field of synthetic biology, heterologous genes are expressed in new host organisms. Similarly, natural or designed proteins are mutagenized at targeted positions and screened for gain-of-function mutations. Here, we describe methods to attain complete randomization or controlled mutations in promoters or genes. Combinatorial libraries of one hundred thousands to tens of millions of variants can be created using commercially synthesized oligonucleotides, simply by performing two rounds of polymerase chain reactions. With a suitably engineered reporter in a whole cell, these libraries can be screened rapidly by performing fluorescence-activated cell sorting (FACS). Within a few rounds of positive and negative sorting based on the response from the reporter, the library can rapidly converge to a few optimal or extremely rare variants with desired phenotypes. Library construction, transformation and sequence verification takes 6–9 days and requires only basic molecular biology lab experience. Screening the library by FACS takes 3–5 days and requires training for the specific cytometer used. Further steps after sorting, including colony picking, sequencing, verification, and characterization of individual clones may take longer, depending on number of clones and required experiments.

59 BASIC BIOLOGICAL SCIENCES↗

Aeolian processes on Venus

This review assesses the potential aeolian regime on Venus as derived from spacecraft observations, laboratory simulations, and theoretical considerations. The two requirements for aeolian processes (a supply of small, loose particles and winds of sufficient strength to move them) appear to be met on Venus. Venera 9, 10, 13, and 14 images show particles considered to be sand and silt size on the surface. In addition, dust spurts (grains 5 to 50 microns in diameter) observed via lander images and inferred from the Pioneer-Venus nephalometer experiments suggest that the particles are loose and subject to movement. Although data on near surface winds are limited, measurements of 0.3 to 1.2 m/sec from the Venera lander and Pioneer-Venus probes appear to be well within the range required for sand and dust entrainment. Aeolian activity involves the interaction of the atmosphere, lithosphere, and loose particles. Thus, there is the potential for various physical and chemical weathering processes that can effect not only rates of erosion, but changes in the composition of all three components. The Venus Simulator is an apparatus used to simulate weathering under venusian conditions at full pressure (to 112 bars) and temperature (to 800 K). In one series of tests, the physical modifications of windblown particles and rock targets were assessed and it was shown that particles become abraded even when moved by gentle winds. However, little abrasion occurs on the target faces. Thus, compositional signatures for target rocks may be more indicative of the windblown particles than of the bedrock. From these and other considerations, aeolian modifications of the venusian surface may be expected to occur as weathering, erosion, transportation, and deposition of surficial materials. Depending upon global and local wind regimes, there may be distinctive sources and sinks of windblown materials. Radar imaging, especially as potentially supplied via the Magellan mission, may enable the identification of such areas on Venus.

Greeley, Ronald↗