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At least 1,225 records · Page 68

Modular protein-oligonucleotide signal exchange

Abstract While many methods are available to measure the concentrations of proteins in solution, the development of a method to quantitatively report both increases and decreases in different protein concentrations in real-time using changes in the concentrations of other molecules, such as DNA outputs, has remained a challenge. Here, we present a biomolecular reaction process that reports the concentration of an input protein in situ as the concentration of an output DNA oligonucleotide strand. This method uses DNA oligonucleotide aptamers that bind either to a specific protein selectively or to a complementary DNA oligonucleotide reversibly using toehold-mediated DNA strand-displacement. It is possible to choose the sequence of output strand almost independent of the sensing protein. Using this strategy, we implemented four different exchange processes to report the concentrations of clinically relevant human α-thrombin and vascular endothelial growth factor using changes in concentrations of DNA oligonucleotide outputs. These exchange processes can operate in tandem such that the same or different output signals can indicate changes in concentration of distinct or identical input proteins. The simplicity of our approach suggests a pathway to build devices that can direct diverse output responses in response to changes in concentrations of specific proteins.

59 BASIC BIOLOGICAL SCIENCES↗

Dynamics of Long-period Comets

Dynamical studies of the origin and evolution of long period comets in the Oort cloud during the past year have concentrated on four areas: (1) interpretation of IRAS observations of dust shells around Vega and some 40 other main sequence stars as evidence for cometary clouds around each of these stars; (2) the dynamical plausibility of an unseen solar companion star orbiting in the Oort cloud and causing periodic cometary showers which result in biological extinction events on the earth; (3) a review of the current hypotheses for cometary formation with particular attention to how each mechanism supplies the required mass of comets to the Oort cloud; and (4) development of new dynamics software to simulate the passage of individual stars directly through the Oort cloud. Each of these efforts is described in detail.

Weissman, P. R.↗

Ceratopteris richardii: a productive model for revealing secrets of signaling and development

Ceratopteris richardii is an aquatic fern grown in tropical and subtropical regions of the world. It is proven to be a productive model system for studies in the genetics, biochemistry, and cell biology of basic biologic processes that occur in early gametophytic development. It provides several advantages to biologists, especially those interested in gravitational biology, polarity development, and in the genetics of sexual development. It is easy to culture, has a relatively short life cycle, and offers an array of attractive features that facilitate genetic studies. The germination and early development of large populations of genetically identical spores are easy to synchronize, and both the direction of polarity development and cell-level gravity responses can be measured and readily manipulated within the first 24 h of spore development. Although there is no reliable transformation system available yet in Ceratopteris, recent studies suggest that the technique of RNA interference can be used to block translation of specific genes in a related fern, Marsilea, and current studies will soon reveal the applicability of this approach, as well as of other transformation approaches, in Ceratopteris. A recently completed expressed sequence tag (EST) sequencing project makes available the partial sequence of more than 2000 cDNAs, representing a significant percentage of the genes being expressed during the first 24 h of spore germination, when many developmentally interesting processes are occurring. A microarray of these ESTs is being constructed, so especially for those scientists interested in basic cellular phenomena that occur early in spore germination, the availability of the ESTs and of the microarray will make Ceratopteris an even more attractive model system.

NASA Discipline Plant Biology↗

Initial Incidence of White Matter Hyperintensities on MRI in Astronauts

Introduction: Previous literature has described the increase in white matter hyperintensity (WMH) burden associated with hypobaric exposure in the U-2 and altitude chamber operating personnel. Although astronauts have similar hypobaric exposure pressures to the U2 pilot population, astronauts have far fewer exposures and each exposure would be associated with a much lower level of decompression stress due to rigorous countermeasures to prevent decompression sickness. Therefore, we postulated that the WMH burden in the astronaut population would be less than in U2 pilots. Methods: Twenty-one post-flight de-identified astronaut MRIs (5 mm slice thickness FLAIR sequences) were evaluated for WMH count and volume. The only additional data provided was an age range of the astronauts (43-57) and if they had ever performed an EVA (13 yes, 8 no). Results: WMH count in these 21 astronaut MRI was 21.0 +/- 24.8 (mean+/- SD) and volume was 0.382 +/- 0.602 ml, which was significantly higher than previously published results for the U2 pilots. No significant differences between EVA and no EVA groups existed. Age range of astronaut population is not directly comparable to the U2 population. Discussion: With significantly less frequent (sometimes none) and less stressful hypobaric exposures, yet a much higher incidence of increased WMH, this indicates the possibility of additional mechanisms beyond hypobaric exposure. This increase unlikely to be attributable just to the differences in age between astronauts and U2 pilots. Forward work includes continuing review of post-flight MRI and evaluation of pre to post flight MRI changes if available. Data mining for potential WMH risk factors includes collection of age, sex, spaceflight experience, EVA hours, other hypobaric exposures, hyperoxic exposures, radiation, high performance aircraft experience and past medical history. Finally, neurocognitive and vision/eye results will be evaluated for any evidence of impairment linked to increased WMH.

Norcross, Jason↗

Direct observation of the exotic β – γ – α decay mode in the T z = – 1 nucleus Na 20

The exotic β – γ – α decay mode of 20 Na has been directly observed for the first time in the Day-one experiment at the Beijing Radioactive Ion-beam Facility. The 20 Na source was produced by using a 100-MeV proton beam bombarding a stack of microporous MgO thick target and delivered as an intense mass separated beam after online ionization. A high-efficiency simultaneous measurement of β, γ, and α transitions enables the β-delayed γ – γ and α – γ coincidence spectroscopy. Here three β – γ – α exotic decay sequences in 20 Na are discovered, which expands the rare decay modes observed in β decay. Moreover, a β – α –decay sequence to the 6130-keV 3 – state of 16 O is observed, which is likely through the 12367-keV 1 + state in 20 Ne. The experimentally deduced B(F) and B(GT) are compared to the shell-model calculation, the allowed β transition strengths can be well accounted for by using sd shell-model space interactions.

73 NUCLEAR PHYSICS AND RADIATION PHYSICS↗

CRISPR/Cas9-Mediated Multi-Allelic Gene Targeting in Sugarcane Confers Herbicide Tolerance

Sugarcane is the source of 80% of the sugar and 26% of the bioethanol produced globally. However, its complex, highly polyploid genome (2n = 100 – 120) impedes crop improvement. Here, we report efficient and reproducible gene targeting (GT) in sugarcane, enabling precise co-editing of multiple alleles via template-mediated and homology-directed repair (HDR) of DNA double strand breaks induced by the programmable nuclease CRISPR/Cas9. The evaluation of 146 independently transformed plants from five independent experiments revealed a targeted nucleotide replacement that resulted in both targeted amino acid substitutions W574L and S653I in the acetolactate synthase (ALS) in 11 lines in addition to single, targeted amino acid substitutions W574L or S653I in 25 or 18 lines, respectively. Co-editing of up to three ALS copies/alleles that confer herbicide tolerance was confirmed by Sanger sequencing of cloned long polymerase chain reaction (PCR) amplicons. This work will enable crop improvement by conversion of inferior alleles to superior alleles through targeted nucleotide substitutions.

Altpeter, Fredy (ORCID:0000000208944976)↗

Neural-network-directed alignment of optical systems using the laser-beam spatial filter as an example

This report describes an effort at NASA Lewis Research Center to use artificial neural networks to automate the alignment and control of optical measurement systems. Specifically, it addresses the use of commercially available neural network software and hardware to direct alignments of the common laser-beam-smoothing spatial filter. The report presents a general approach for designing alignment records and combining these into training sets to teach optical alignment functions to neural networks and discusses the use of these training sets to train several types of neural networks. Neural network configurations used include the adaptive resonance network, the back-propagation-trained network, and the counter-propagation network. This work shows that neural networks can be used to produce robust sequencers. These sequencers can learn by example to execute the step-by-step procedures of optical alignment and also can learn adaptively to correct for environmentally induced misalignment. The long-range objective is to use neural networks to automate the alignment and operation of optical measurement systems in remote, harsh, or dangerous aerospace environments. This work also shows that when neural networks are trained by a human operator, training sets should be recorded, training should be executed, and testing should be done in a manner that does not depend on intellectual judgments of the human operator.

Decker, Arthur J.↗

Developing a Standard Method for Link-Layer Security of CCSDS Space Communications

Communications security for space systems has been a specialized field generally far removed from considerations of mission interoperability and cross-support in fact, these considerations often have been viewed as intrinsically opposed to security objectives. The space communications protocols defined by the Consultative Committee for Space Data Systems (CCSDS) have a twenty-five year history of successful use in over 400 missions. While the CCSDS Telemetry, Telecommand, and Advancing Orbiting Systems protocols for use at OSI Layer 2 are operationally mature, there has been no direct support within these protocols for communications security techniques. Link-layer communications security has been successfully implemented in the past using mission-unique methods, but never before with an objective of facilitating cross-support and interoperability. This paper discusses the design of a standard method for cryptographic authentication, encryption, and replay protection at the data link layer that can be integrated into existing CCSDS protocols without disruption to legacy communications services. Integrating cryptographic operations into existing data structures and processing sequences requires a careful assessment of the potential impediments within spacecraft, ground stations, and operations centers. The objective of this work is to provide a sound method for cryptographic encapsulation of frame data that also facilitates Layer 2 virtual channel switching, such that a mission may procure data transport services as needed without involving third parties in the cryptographic processing, or split independent data streams for separate cryptographic processing.

Biggerstaff, Craig↗

Cloudless Atmospheres for Young Low-Gravity Substellar Objects

Atmospheric modeling of low-gravity (VL-G) young brown dwarfs remains challenging. The presence of very thick clouds is a possible source of this challenge, because of their extremely red near-infrared (NIR) spectra, but no cloud models provide a good fit to the data with a radius compatible with the evolutionary models for these objects. We show that cloudless atmospheres assuming a temperature gradient reduction caused by fingering convection provide a very good model to match the observed VL-G NIR spectra. The sequence of extremely red colors in the NIR for atmospheres with effective temperatures from ∼2000 K down to ∼1200 K is very well reproduced with predicted radii typical of young low-gravity objects. Future observations with NIRSPEC and MIRI on the James Webb Space Telescope (JWST) will provide more constraints in the mid-infrared, helping to confirm or refute whether or not the NIR reddening is caused by fingering convection. We suggest that the presence or absence of clouds will be directly determined by the silicate absorption features that can be observed with MIRI. JWST will therefore be able to better characterize the atmosphere of these hot young brown dwarfs and their low-gravity exoplanet analogs.

Tremblin, P.↗

Vertical Habitability Layout Studies and Neutral Buoyancy/Parabolic Flight Habitat Studies

In the Summer of 2013, the University of Maryland was selected for two projects under the NASA X-Hab 2014 program, administered by the National Space Grant Foundation: Vertical Habitability Layout Studies (XHab201406) and Neutral Buoyancy/Parabolic Flight Habitat Studies (XHab201407). This document, at the direction of the NASA Technical Monitor, comprises the final report for both of these contracts. Recognizing from the outset the mismatch between the desired scope of research activities under the contracts and the severely limited funding and duration, the University of Maryland leveraged the X-Hab support by integrating the programs into a number of academic classes throughout the 2013-2014 academic year. By far the most significant interaction was with the Department of Aerospace Engineering Senior capstone experience in spacecraft design sequence, ENAE 483/484. Throughout the academic year, 42 students in this sequence worked on both projects in conjunction with their senior project to design an artificial gravity research station in a distant lunar retrograde orbit. As part of this research project, the students worked with previously created 1-G habitats and underwater simulations to better understand habitat design from microgravity to full Earth gravity, as well as at lunar and Mars gravity levels between those two endpoints.

David L. Akin↗

The ram accelerator - A chemically driven mass launcher

The ram accelerator, a chemically propelled mass driver, is presented as a viable new approach for directly launching acceleration-insensitive payloads into low earth orbit. The propulsion principle is similar to that of a conventional air-breathing ramjet. The cargo vehicle resembles the center-body of a ramjet and travels through a tube filled with a pre-mixed fuel and oxidizer mixture. The launch tube acts as the outer cowling of the ramjet and the combustion process travels with the vehicle. Two drive modes of the ram accelerator propulsion system are described, which when used in sequence are capable of accelerating the vehicle to as high as 10 km/sec. The requirements are examined for placing a 2000 kg vehicle into a 500 km orbit with a minimum of on-board rocket propellant for circularization maneuvers. It is shown that aerodynamic heating during atmospheric transit results in very little ablation of the nose. An indirect orbital insertion scenario is selected, utilizing a three step maneuver consisting of two burns and aerobraking. An on-board propulsion system using storable liquid propellants is chosen in order to minimize propellant mass requirements, and the use of a parking orbit below the desired final orbit is suggested as a means to increase the flexibility of the mass launch concept. A vehicle design using composite materials is proposed that will best meet the structural requirements, and a preliminary launch tube design is presented.

Kaloupis, P.↗

Programmable display pushbuttons on the Space Station's telerobot control panel

The Man-Systems Telerobotics Laboratory at NASA's Johnson Space Center and supported by Lockheed, is working to ensure that the Flight Telerobotic Servicer (FTS) to be used on the Space Shuttle (Orbiter) and the Space Station has a well designed user interface from a Human Factors perspective. The FTS, which is a project led by NASA's Goddard Space Flight Center, will be a telerobot used for Space Station construction, maintenance, and satellite repair. It will be directly controlled from workstations on the Orbiter and the Space Station and monitored from a ground workstation. The FTS will eventually evolve into a more autonomous system, but in the short-term the system will be manually operated (teleoperated) for many tasks. This emphasizes the importance of the human/telerobot interface on this system. This phase of the FTS workstation evaluation covers a preliminary study of programmable display pushbuttons (PDP's). The PDP is constructed of a matrix of directly addressable electroluminescent (EL) pixels which can be used to form dot-matrix characters. PDP's can be used to display more than one message and to control more than one function. Since the PDP's have these features, then a single PDP may possibly replace the use of many single-function pushbuttons, rotary switches, and toggle switches, thus using less panel space. It is of interest to determine if PDP's can be used to adequately perform complex hierarchically structured task sequences. The objective of this investigation was to compare the performance of experienced and inexperienced Remote Manipulator System (RMS) operators while performing an RMS like task on simulated PDP and non-PDP computer prototypes so that guidelines governing the use of programmable display pushbuttons on the FTS workstation could be created. The functionality of the RMS on the Orbiter was used as a model for this evaluation since the functionality of the FTS at the time of this writing has not been solidified.

Stuart, Mark A.↗

Simulation of Peptides at Aqueous Interfaces

Behavior of peptides at water-membrane interfaces is of great interest in studies on cellular transport and signaling, membrane fusion, and the action of toxins and antibiotics. Many peptides, which exist in water only as random coils, can form sequence-dependent, ordered structures at aqueous interfaces, incorporate into membranes and self-assembly into functional units, such as simple ion channels. Multi -nanosecond molecular dynamics simulations have been carried out to study the mechanism and energetics of interfacial folding of both non-polar and amphiphilic peptides, their insertion into membranes and association into higher-order structures. The simulations indicate that peptides fold non-sequentially, often through a series of amphiphilic intermediates. They further incorporate into the membrane in a preferred direction as folded monomers, and only then aggregate into dimers and, possibly, further into "dimers of dimers".

Pohorille, Andrew↗

The Arabidopsis SKU5 gene encodes an extracellular glycosyl phosphatidylinositol-anchored glycoprotein involved in directional root growth

To investigate how roots respond to directional cues, we characterized a T-DNA-tagged Arabidopsis mutant named sku5 in which the roots skewed and looped away from the normal downward direction of growth on inclined agar surfaces. sku5 roots and etiolated hypocotyls were slightly shorter than normal and exhibited a counterclockwise (left-handed) axial rotation bias. The surface-dependent skewing phenotype disappeared when the roots penetrated the agar surface, but the axial rotation defect persisted, revealing that these two directional growth processes are separable. The SKU5 gene belongs to a 19-member gene family designated SKS (SKU5 Similar) that is related structurally to the multiple-copper oxidases ascorbate oxidase and laccase. However, the SKS proteins lack several of the conserved copper binding motifs characteristic of copper oxidases, and no enzymatic function could be assigned to the SKU5 protein. Analysis of plants expressing SKU5 reporter constructs and protein gel blot analysis showed that SKU5 was expressed most strongly in expanding tissues. SKU5 was glycosylated and modified by glycosyl phosphatidylinositol and localized to both the plasma membrane and the cell wall. Our observations suggest that SKU5 affects two directional growth processes, possibly by participating in cell wall expansion.

NASA Discipline Plant Biology↗

Evaluation of the Performance Characteristics of the CGLSS and NLDN Systems Based on Two Years of Ground-Truth Data from Launch Complex 39B, Kennedy Space Center, Florida

From May 2011 through July 2013, the lightning instrumentation at Launch Complex 39B (LC39B) at the Kennedy Space Center, Florida, has obtained high-speed video records and field change waveforms (dE/dt and three-axis dH/dt) for 54 negative polarity return strokes whose strike termination locations and times are known with accuracy of the order of 10 m or less and 1 μs, respectively. A total of 18 strokes terminated directly to the LC39B lighting protection system (LPS), which contains three 181 m towers in a triangular configuration, an overhead catenary wire system on insulating masts, and nine down conductors. An additional 9 strokes terminated on the 106 m lightning protection mast of Launch Complex 39A (LC39A), which is located about 2.7 km southeast of LC39B. The remaining 27 return strokes struck either on the ground or attached to low-elevation grounded objects within about 500 m of the LC39B LPS. Leader/return stroke sequences were imaged at 3200 frames/sec by a network of six Phantom V310 high-speed video cameras. Each of the three towers on LC39B had two high-speed cameras installed at the 147 m level with overlapping fields of view of the center of the pad. The locations of the strike points of 54 return strokes have been compared to time-correlated reports of the Cloud-to-Ground Lightning Surveillance System (CGLSS) and the National Lightning Detection Network (NLDN), and the results of this comparison will be presented and discussed.

Waveforms↗

Neutralization profiles of HIV-1 viruses from the VRC01 Antibody Mediated Prevention (AMP) trials

The VRC01 Antibody Mediated Prevention (AMP) efficacy trials conducted between 2016 and 2020 showed for the first time that passively administered broadly neutralizing antibodies (bnAbs) could prevent HIV-1 acquisition against bnAb-sensitive viruses. HIV-1 viruses isolated from AMP participants who acquired infection during the study in the sub-Saharan African (HVTN 703/HPTN 081) and the Americas/European (HVTN 704/HPTN 085) trials represent a panel of currently circulating strains of HIV-1 and offer a unique opportunity to investigate the sensitivity of the virus to broadly neutralizing antibodies (bnAbs) being considered for clinical development. Pseudoviruses were constructed using envelope sequences from 218 individuals. The majority of viruses identified were clade B and C; with clades A, D, F and G and recombinants AC and BF detected at lower frequencies. We tested eight bnAbs in clinical development (VRC01, VRC07-523LS, 3BNC117, CAP256.25, PGDM1400, PGT121, 10–1074 and 10E8v4) for neutralization against all AMP placebo viruses (n = 76). Compared to older clade C viruses (1998–2010), the HVTN703/HPTN081 clade C viruses showed increased resistance to VRC07-523LS and CAP256.25. At a concentration of 1μg/ml (IC80), predictive modeling identified the triple combination of V3/V2-glycan/CD4bs-targeting bnAbs (10-1074/PGDM1400/VRC07-523LS) as the best against clade C viruses and a combination of MPER/V3/CD4bs-targeting bnAbs (10E8v4/10-1074/VRC07-523LS) as the best against clade B viruses, due to low coverage of V2-glycan directed bnAbs against clade B viruses. Overall, the AMP placebo viruses represent a valuable resource for defining the sensitivity of contemporaneous circulating viral strains to bnAbs and highlight the need to update reference panels regularly. Our data also suggests that combining bnAbs in passive immunization trials would improve coverage of global viruses.

60 APPLIED LIFE SCIENCES↗

Metagenomic Characterization of Soil Microbial Communities in the Luquillo Experimental Forest (Puerto Rico) and Implications for Nitrogen Cycling

The phylogenetic and functional diversities of microbial communities in tropical rainforests and how these differ from those of temperate communities remain poorly described but are directly related to the increased fluxes of greenhouse gases such as nitrous oxide (N 2 O) from the tropics. Toward closing these knowledge gaps, in this study we analyzed replicated shotgun metagenomes representing distinct life zones and an elevation gradient from four locations in the Luquillo Experimental Forest (LEF), Puerto Rico. These soils had a distinct microbial community composition and lower species diversity compared to those of temperate grasslands or agricultural soils. In contrast to the overall distinct community composition, the relative abundances and nucleotide sequences of N 2 O reductases (nosZ) were highly similar between tropical forest and temperate soils. However, respiratory NO reductase (norB) was 2-fold more abundant in the tropical soils, which might be relatable to their greater N 2 O emissions. Nitrogen fixation (nifH) also showed higher relative abundance in rainforest than in temperate soils, i.e., 20% versus 0.1 to 0.3% of bacterial genomes in each soil type harbored the gene, respectively. Finally, unlike temperate soils, LEF soils showed little stratification with depth in the first 0 to 30 cm, with ~45% of community composition differences explained solely by location. Collectively, these results advance our understanding of spatial diversity and metabolic repertoire of tropical rainforest soil communities and should facilitate future ecological studies of these ecosystems.

54 ENVIRONMENTAL SCIENCES↗

Landmark Publications in Analytical Atomic Spectrometry: Fundamentals and Instrumentation Development

The almost-two-centuries history of spectrochemical analysis has generated a body of literature so vast that it has become nearly intractable for experts, much less for those wishing to enter the field. Authoritative, focused reviews help to address this problem but become so granular that the overall directions of the field are lost. This broader perspective can be provided partially by general overviews but then the thinking, experimental details, theoretical underpinnings, and instrumental innovations of the original work must be sacrificed. In the present compilation, this dilemma is overcome by assembling the most impactful publications in the area of analytical atomic spectrometry. Each entry was proposed by at least one current expert in the field and supported by a narrative that justifies its inclusion. The entries were then assembled into a coherent sequence and returned to contributors for a round-robin review. A total of 48 scientists participated in this endeavor, contributing a combined list of 1055 individual articles spanning 17 sub-disciplines of spectrochemical analysis into what the current community views as “key” publications. Of these cited articles, 60 received nominations from four or more scientists, establishing them as the most indispensable reading materials. The outcome of this collaborative effort is intended to serve as a valuable resource not only for current practitioners in atomic spectroscopy but also for present and future students who represent coming generations of analytical atomic spectroscopists.

Instruments & Instrumentation↗