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At least 1,153 records · Page 64

Data for Development, Optimization, and Application of an Episomal Plasmid System for Rhodotorula toruloides

Rhodotorula toruloides is an emerging oleaginous yeast with strong potential as a microbial cell factory for the production of acetyl-CoA-derived bioproducts. However, engineering of this organism has been limited by the absence of a functional episomal plasmid system, a foundational genetic tool for rapid gene expression, pathway testing, and CRISPR-based genome engineering. Here, we report the first episomal plasmid system for R. toruloides . Through systematic screening of candidate autonomously replicating sequences (ARSs) from diverse sources, we identified multiple functional ARS elements and selected C63F4, a fragment derived from Contig 63 of R. toruloides CBS14, because of its stable performance. The resulting pC63F4 plasmid was maintained episomally, supported GFP reporter expression, exhibited a copy number of 2.39 ± 0.13, and showed good stability during long term cultivation. To overcome poor transformation efficiency, we developed a Cre-loxP-mediated in vivo re-circularization strategy that enabled reliable delivery of the episomal plasmid. Using this improved system, we demonstrated functional episomal expression of metabolic engineering genes and multi-gene pathways for the production of triacetic acid lactone, fatty alcohols, and limonene. Finally, we leveraged this platform to establish a redesigned CRISPR system that enables seamless genome editing in R. toruloides for the first time, while also simplifying marker recycling. Together, this work establishes a long-needed episomal plasmid platform and associated CRISPR toolkit that will accelerate metabolic engineering, synthetic biology, and fundamental studies in R. toruloides .

Gene Editing↗

Apollo Operations Handbook Lunar Module (LM 11 and Subsequent) Vol. 2 Operational Procedures

The Apollo Operations Handbook (AOH) is the primary means of documenting LM descriptions and procedures. The AOH is published in two separately bound volumes. This information is useful in support of program management, engineering, test, flight simulation, and real time flight support efforts. This volume contains crew operational procedures: normal, backup, abort, malfunction, and emergency. These procedures define the sequence of actions necessary for safe and efficient subsystem operation.

Source record↗

Growth of surface and corner cracks in beta-processed and mill-annealed Ti-6Al-4V

Empirical stress-intensity expressions were developed to relate the growth of cracks from corner flaws to the growth of cracks from surface flaws. An experimental program using beta-processed Ti-6Al-4V verified these expressions for stress ratios, R greater than or equal to 0. An empirical crack growth-rate expression which included stress-ratio and stress-level effects was also developed. Cracks grew approximately 10 percent faster in transverse-grain material than in longitudinal-grain material and at approximately the same rate in longitudinal-grain mill-annealed Ti-6Al-4V. Specimens having surface and corner cracks and made of longitudinal-grain, beta-processed material were tested with block loads, and increasing the stresses in a block did not significantly change the crack growth rates. Truncation of the basic ascending stress sequence within a block caused more rapid crack growth, whereas both the descending and low-to-high stress sequences slowed crack growth.

Bell, P. D.↗

Structural features of the nonionic surfactants stabilizing long-lived bubble nuclei

A study of the effects of various electrolytes and one organic compound on bubble production in agarose gels is presented. Several preparations of ultrapure agarose were compared for 42 electrolytes and phenol to identify trends in bubble formation. The anion and cation sequences of bubble suppression are similar to processes for salting out of nonionic surfactants. The reduction of bubble number by polyvalent ions and 1% phenol suggests that the polar portions of these nonionic surfactants represent amide groups. The evidence for amide groups is consistent with the relative positions of Mg(2+) in all cation sequences; this result makes it unlikely that either linkages contribute to the hydrophilicity of the nonionic surfactants stabilizing bubble nuclei in the different aqueous gels tested.

Darrigo, J. S.↗

Satellite Emission Range Inferred Earth Survey (SERIES) project

The Global Positioning System (GPS) was developed by the Department of Defense primarily for navigation use by the United States Armed Forces. The system will consist of a constellation of 18 operational Navigation Satellite Timing and Ranging (NAVSTAR) satellites by the late 1980's. During the last four years, the Satellite Emission Range Inferred Earth Surveying (SERIES) team at the Jet Propulsion Laboratory (JPL) has developed a novel receiver which is the heart of the SERIES geodetic system designed to use signals broadcast from the GPS. This receiver does not require knowledge of the exact code sequence being transmitted. In addition, when two SERIES receivers are used differentially to determine a baseline, few cm accuracies can be obtained. The initial engineering test phase has been completed for the SERIES Project. Baseline lengths, ranging from 150 meters to 171 kilometers, have been measured with 0.3 cm to 7 cm accuracies. This technology, which is sponsored by the NASA Geodynamics Program, has been developed at JPL to meet the challenge for high precision, cost-effective geodesy, and to complement the mobile Very Long Baseline Interferometry (VLBI) system for Earth surveying.

Buennagel, L. A.↗

The Final Approach Spacing Tool (FAST)

The Final Approach Spacing Tool (FAST) is a decision support tool for terminal area (TRACON) air traffic controllers. FAST utilizes 4D trajectory synthesis, human performance modeling, and a graphical user interface to plan and advise efficient, conflict-free aircraft trajectories for arrival traffic. It increases airport capacity, reduces arrival delays, and reduces controller workload by issuing: sequence and runway advisories ("Passive" FAST) and speed and heading advisories ("Active" FAST). This paper discusses in detail the architecture of the system and operational tests done on the system.

Slattery, Rhonda↗

Game-like tasks for comparative research: leveling the playing field

Game-like computer tasks offer many benefits for psychological research. In this paper, the usefulness of such tasks to bridge population differences (e.g., age, intelligence, species) is discussed and illustrated. A task called ALVIN was used to assess humans' and monkeys' working memory for sequences of colors with or without tones. Humans repeated longer lists than did the monkeys, and only humans benefited when the visual stimuli were accompanied by auditory cues. However, the monkeys did recall sequences at levels comparable to those reported elsewhere for children. Comparison of similarities and differences between the species is possible because the two groups were tested with exactly the same game-like paradigm.

NASA Discipline Number 06-10↗

Reefing Line Tension in CPAS Main Parachute Clusters

Reefing lines are an essential feature to manage inflation loads. During each Engineering Development Unit (EDU) test of the Capsule Parachute Assembly System (CPAS), a chase aircraft is staged to be level with the cluster of Main ringsail parachutes during the initial inflation and reefed stages. This allows for capturing high-quality still photographs of the reefed skirt, suspension line, and canopy geometry. The over-inflation angles are synchronized with measured loads data in order to compute the tension force in the reefing line. The traditional reefing tension equation assumes radial symmetry, but cluster effects cause the reefed skirt of each parachute to elongate to a more elliptical shape. This effect was considered in evaluating multiple parachutes to estimate the semi-major and semi-minor axes. Three flight tests are assessed, including one with a skipped first stage, which had peak reefing line tension over three times higher than the nominal parachute disreef sequence.

Ray, Eric S.↗

Alignment Test Results of the JWST Pathfinder Telescope Mirrors in the Cryogenic Environment

After integration of the Optical Telescope Element (OTE) to the Integrated Science Instrument Module (ISIM) to become the OTIS, the James Webb Space Telescope OTIS is tested at NASAs Johnson Space Center (JSC) in the cryogenic vacuum Chamber A for alignment and optical performance. The alignment of the mirrors comprises a sequence of steps as follows: The mirrors are coarsely aligned using photogrammetry cameras with reflective targets attached to the sides of the mirrors. Then a multi-wavelength interferometer is aligned to the 18-segment primary mirror using cameras at the center of curvature to align reflected light from the segments and using fiducials at the edge of the primary mirror. Once the interferometer is aligned, the 18 primary mirror segments are then adjusted to optimize wavefront error of the aggregate mirror. This process phases the piston and tilt positions of all the mirror segments. An optical fiber placed at the Cassegrain focus of the telescope then emits light towards the secondary mirror to create a collimated beam emitting from the primary mirror. Portions of the collimated beam are retro-reflected from flat mirrors at the top of the chamber to pass through the telescope to the SI detector. The image on the detector is used for fine alignment of the secondary mirror and a check of the primary mirror alignment using many of the same analysis techniques used in the on-orbit alignment. The entire process was practiced and evaluated in 2015 at cryogenic temperature with the Pathfinder telescope.

Interferometer↗

Measuring and Modeling Shared Visual Attention

Multi-person teams are sometimes responsible for critical tasks, such as flying an airliner. Here we present a method using gaze tracking data to assess shared visual attention, a term we use to describe the situation where team members are attending to a common set of elements in the environment. Gaze data are quantized with respect to a set of N areas of interest (AOIs); these are then used to construct a time series of N dimensional vectors, with each vector component representing one of the AOIs, all set to 0 except for the component corresponding to the currently fixated AOI, which is set to 1. The resulting sequence of vectors can be averaged in time, with the result that each vector component represents the proportion of time that the corresponding AOI was fixated within the given time interval. We present two methods for comparing sequences of this sort, one based on computing the time-varying correlation of the averaged vectors, and another based on a chi-square test testing the hypothesis that the observed gaze proportions are drawn from identical probability distributions.We have evaluated the method using synthetic data sets, in which the behavior was modeled as a series of activities, each of which was modeled as a first-order Markov process. By tabulating distributions for pairs of identical and disparate activities, we are able to perform a receiver operating characteristic (ROC) analysis, allowing us to choose appropriate criteria and estimate error rates.We have applied the methods to data from airline crews, collected in a high-fidelity flight simulator (Haslbeck, Gontar Schubert, 2014). We conclude by considering the problem of automatic (blind) discovery of activities, using methods developed for text analysis.

attention↗

Measuring and Modeling Shared Visual Attention

Multi-person teams are sometimes responsible for critical tasks, such as flying an airliner. Here we present a method using gaze tracking data to assess shared visual attention, a term we use to describe the situation where team members are attending to a common set of elements in the environment. Gaze data are quantized with respect to a set of N areas of interest (AOIs); these are then used to construct a time series of N dimensional vectors, with each vector component representing one of the AOIs, all set to 0 except for the component corresponding to the currently fixated AOI, which is set to 1. The resulting sequence of vectors can be averaged in time, with the result that each vector component represents the proportion of time that the corresponding AOI was fixated within the given time interval. We present two methods for comparing sequences of this sort, one based on computing the time-varying correlation of the averaged vectors, and another based on a chi-square test testing the hypothesis that the observed gaze proportions are drawn from identical probability distributions. We have evaluated the method using synthetic data sets, in which the behavior was modeled as a series of "activities," each of which was modeled as a first-order Markov process. By tabulating distributions for pairs of identical and disparate activities, we are able to perform a receiver operating characteristic (ROC) analysis, allowing us to choose appropriate criteria and estimate error rates. We have applied the methods to data from airline crews, collected in a high-fidelity flight simulator (Haslbeck, Gontar & Schubert, 2014). We conclude by considering the problem of automatic (blind) discovery of activities, using methods developed for text analysis.

attention↗

Alignment Test Results of the JWST Pathfinder Telescope Mirrors in the Cryogenic Environment

After integration of the Optical Telescope Element (OTE) to the Integrated Science Instrument Module (ISIM) to become the OTIS, the James Webb Space Telescope OTIS is tested at NASAs Johnson Space Center (JSC) in the cryogenic vacuum Chamber A for alignment and optical performance. The alignment of the mirrors comprises a sequence of steps as follows: The mirrors are coarsely aligned using photogrammetry cameras with reflective targets attached to the sides of the mirrors. Then a multi-wavelength interferometer is aligned to the 18-segment primary mirror using cameras at the center of curvature to align reflected light from the segments and using fiducials at the edge of the primary mirror. Once the interferometer is aligned, the 18 primary mirror segments are then adjusted to optimize wavefront error of the aggregate mirror. This process phases the piston and tilt positions of all the mirror segments. An optical fiber placed at the Cassegrain focus of the telescope then emits light towards the secondary mirror to create a collimated beam emitting from the primary mirror. Portions of the collimated beam are retro-reflected from flat mirrors at the top of the chamber to pass through the telescope to the SI detector. The image on the detector is used for fine alignment of the secondary mirror and a check of the primary mirror alignment using many of the same analysis techniques used in the on-orbit alignment. The entire process was practiced and evaluated in 2015 at cryogenic temperature with the Pathfinder telescope.

OTIS↗

Scheduling Improvements Following the Phase 1 Field Evaluation of the ATD-2 Integrated Arrival, Departure, and Surface Concept

NASA is conducting the Airspace Technology Demonstration-2 to evaluate an Integrated Arrival, Departure, and Surface (IADS) traffic management system that extends traffic sequencing for the entire life-cycle of a flight from departure gate to arrival gate within multi-airport, metroplex environments. After development and testing in human-in-the-loop simulations, the IADS system was deployed to Charlotte Douglas International Airport for a three-year field evaluation. From the initial IADS concept development through the end of the Phase 1 field evaluation many lessons were learned with regards to the IADS scheduler. In this paper we describe how data from the Phase 1 field evaluation helped identify scheduler improvements and guided the implementation of refinements. The improvements in the IADS scheduler described in this paper are incorporated into the IADS Phase 2 scheduler enabling strategic Surface Metering Programs and will be evaluated during the field evaluation.

Coupe, William J.↗

Scheduling Improvements Following the Phase 1 Field Evaluation of the ATD-2 Integrated Arrival, Departure, and Surface Concept

NASA is conducting the Airspace Technology Demonstration-2 (ATD-2) to evaluate an Integrated Arrival, Departure, and Surface (IADS) traffic management system that extends traffic sequencing for the entire life-cycle of a flight from departure gate to arrival gate within multi-airport, metroplex environments. After development and testing in human-in-the-loop simulations, the IADS system was deployed to Charlotte Douglas International Airport for a three-year field evaluation. From the initial IADS concept development through the end of the Phase 1 field evaluation many lessons were learned with regards to the IADS scheduler. In this paper we describe how data from the Phase 1 field evaluation helped identify scheduler improvements and guided the implementation of refinements. The improvements in the IADS scheduler described in this paper are incorporated into the IADS Phase 2 scheduler enabling strategic Surface Metering Programs and will be evaluated during the field evaluation.

Airspace Technology Demonstration-2↗

Ares I-X Flight Test Philosophy

In response to the Vision for Space Exploration, the National Aeronautics and Space Administration (NASA) has defined a new space exploration architecture to return humans to the Moon and prepare for human exploration of Mars. One of the first new developments will be the Ares I Crew Launch Vehicle (CLV), which will carry the Orion Crew Exploration Vehicle (CEV), into Low Earth Orbit (LEO) to support International Space Station (ISS) missions and, later, support lunar missions. As part of Ares I development, NASA will perform a series of Ares I flight tests. The tests will provide data that will inform the engineering and design process and verify the flight hardware and software. The data gained from the flight tests will be used to certify the new Ares/Orion vehicle for human space flight. The primary objectives of this first flight test (Ares I-X) are the following: Demonstrate control of a dynamically similar integrated Ares CLV/Orion CEV using Ares CLV ascent control algorithms; Perform an in-flight separation/staging event between an Ares I-similar First Stage and a representative Upper Stage; Demonstrate assembly and recovery of a new Ares CLV-like First Stage element at Kennedy Space Center (KSC); Demonstrate First Stage separation sequencing, and quantify First Stage atmospheric entry dynamics and parachute performance; and Characterize the magnitude of the integrated vehicle roll torque throughout the First Stage (powered) flight. This paper will provide an overview of the Ares I-X flight test process and details of the individual flight tests.

Davis, S. R.↗

Flow Visualization of Liquid Hydrogen Line Chilldown Tests

We present experimental measurements of wall and fluid temperature during chill-down tests of a warm cryogenic line with liquid hydrogen. Synchronized video and fluid temperature measurements are used to interpret stream temperature profiles versus time. When cold liquid hydrogen starts to flow into the warm line, a sequence of flow regimes, spanning from all-vapor at the outset to bubbly with continuum liquid at the end can be observed at a location far downstream of the cold inlet. In this paper we propose interpretations to the observed flow regimes and fluid temperature histories for two chilldown methods, viz. trickle (i.e. continuous) flow and pulse flow. Calculations of heat flux from the wall to the fluid versus wall temperature indicate the presence of the transition/nucleate boiling regimes only. The present tests, run at typical Reynolds numbers of approx O(10 (exp 5)), are in sharp contrast to similar tests conducted at lower Reynolds numbers where a well-defined film boiling region is observed.

Line chill down↗

The rapidly oscillating Ap stars as a test of stellar chromospheric heating mechanisms

The discovery of a new class of magnetic stars, the rapidly pulsating Ap stars, provides an unprecedented tool for the study of Alfvenic heating in the outer atmospheres of main sequence stars. In this note, it is noted that detection of rotationally-modulated chromospheric emission line fluxes, especially the Ca II, Mg II, and C IV lines, can provide a novel method for investigating the role of Alfvenic heating in stellar atmospheres.

Shore, S. N.↗

Functional characterization of a cellulose synthase, CtCESA1, from the marine red alga Calliarthron tuberculosum (Corallinales)

In land plants and algae, cellulose is important for strengthening cell walls and preventing breakage due to physical forces. Though our understanding of cellulose production by cellulose synthases (CESAs) has seen significant advances for several land plant and bacterial species, functional characterization of this fundamental protein is absent in red algae. Here we identify CESA gene candidates in the calcifying red alga Calliarthron tuberculosum using sequence similarity-based approaches, and elucidate their phylogenetic relationship with other CESAs from diverse taxa. One gene candidate, CtCESA1, was closely related to other putative red algal CESA genes. To test if CtCESA1 encoded a true cellulose synthase, CtCESA1 protein was expressed and purified from insect and yeast expression systems. CtCESA1 showed glucan synthase activity in glucose tracer assays. CtCESA1 activity was relatively low when compared with plant and bacterial CESA activity. In an in vitro assay, a predicted N-terminal starch-binding domain from CtCESA1 bound red algal floridean starch extracts, representing a unique domain in red algal CESAs not present in CESAs from other lineages. When the CtCESA1 gene was introduced into Arabidopsis thaliana cesa mutants, the red algal CtCESA1 partially rescued the growth defects of the primary cell wall cesa6 mutant, but not cesa3 or secondary cell wall cesa7 mutants. A fluorescently tagged CtCESA1 localized to the plasma membrane in the Arabidopsis cesa6 mutant background. This study presents functional evidence validating the sequence annotation of red algal CESAs. Furthermore, the relatively low activity of CtCESA1, partial complementation in Arabidopsis, and presence of unique protein domains suggest that there are probably functional differences between the algal and land plant CESAs.

59 BASIC BIOLOGICAL SCIENCES↗