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Method Development for In-situ Detection of Latent Herpesvirus DNA from Saliva using Nanopore Sequencing

Research toward latent herpesvirus reactivation has been intensively addressed through Space Shuttle and International Space Station (ISS) investigations. This work has provided the understanding that persistent reactivation of herpesviruses from asymptomatic crew can be detected through viral shedding in saliva, urine, and blood. Occasionally, viral reactivation from the latency stage can pose a threat to crew health (clinical manifestation) before, during, and after flight missions. Furthermore, previous work detailing correlations with immunity indicate that monitoring viral reactivation could be implemented to assess potential immune dysfunction. While in-flight monitoring is desirable, there is no well-established procedure or method for real-time evaluations, and research to date has relied on postflight, ground-based analysis. The development of portable molecular technologies like the miniPCR™ (miniPCR Bio) thermal cycler and the MinION™ sequencer (Oxford Nanopore Technologies) have confirmed that real-time monitoring is possible in extreme and low resource environments. These devices, combined with simple sample preparation methods, have been used to demonstrate bacterial identification onboard the ISS, as well as rapid viral detection in remote locations on Earth. The work here builds upon previous molecular advancements onboard the ISS toward the development and validation of a spaceflight-compatible method for viral detection from crew samples. Several herpesviruses can be detected in saliva, which provides a non-invasive means to collect samples for monitoring. While the basis for this method lies in previous spaceflight investigations, key points for method optimization include DNA extraction from saliva, viral primer selection, and bioinformatic processes for data analysis. To increase viral yield, numerous DNA extraction methods have been evaluated and will be discussed in detail. For initial development and testing, varicella-zoster virus (VZV) is being targeted though open reading frame 51 and 63 (ORF51, ORF63), as the replication origin-binding protein is highly expressed during latency. Optimization of the thermal cycling parameters has resulted in the ability to test the entire process. The full method has been tested with both viral VZV DNA standards and saliva spiked with varying concentrations of VZV. Viral sequence data were mapped to the reference sequence using minimap2. Prior to mapping, DNA sequencing reads were filtered for length and quality, barcodes were removed, and alignment identity calculated. Following further assessments, statistics were compared across multiple sequencing experiments and are being used to determine the success of the protocols. Forward work will include the incorporation of herpes simplex virus 1 (HSV-1) and Epstein-Barr virus (EBV) primers as well as the validation of results to the terrestrial qPCR standard assay. Upon full validation of the developed method, saliva will be collected from 20 healthy subjects and spiked with viral DNA. These samples will be split and assayed with the MinION and standard qPCR assay.

Hang N Nguyen

Resistivity due to weak double layers - A model for auroral arc thickness

We have calculated the resistivity due to a sequence of fluctuating weak double layers aligned parallel to the ambient magnetic field line. The average response of an electron drifting through a 1D randomly oriented array of WDLs is studied using a test particle approach. The average is taken over the randomly fluctuating values of the electric field associated with the double layers. Based on our calculations, we estimate that a 350 eV electron energy the thickness of the visual auroral arc is about 2.5 km and that of the auroral fine structure as about 250 m when mapped down to the ionosphere. The significance of our calculations is discussed in the context of magnetosphere-ionosphere coupling.

Prakash, Manju

High precision electric gate for time-of-flight ion mass spectrometers

A time-of-flight mass spectrometer having a chamber with electrodes to generate an electric field in the chamber and electric gating for allowing ions with a predetermined mass and velocity into the electric field. The design uses a row of very thin parallel aligned wires that are pulsed in sequence so the ion can pass through the gap of two parallel plates, which are biased to prevent passage of the ion. This design by itself can provide a high mass resolution capability and a very precise start pulse for an ion mass spectrometer. Furthermore, the ion will only pass through the chamber if it is within a wire diameter of the first wire when it is pulsed and has the right speed so it is near all other wires when they are pulsed.

Sittler, Edward C.

TA-13: Ground and Launch Systems, 2015 NASA Technology Roadmaps

This presentation is a summary of new content contained in the 2015 update of Technology Area-13, Ground and Launch Systems technology roadmap beyond the content contained in the 2010 version. Also included are brief assessments of benefits, alignments, challenges, technical risk and reasonableness, sequencing and timing, and time and effort to achieve goals. This presentation is part of overall presentations of new content only for the 2015 update of the 15 NASA Technology Roadmaps that will be conducted in a public forum managed by the National Research Council on September 28-29, 2015. The 15 roadmaps have already been publically released via the STI process.

decision-making tools

Method and apparatus for disabling an echo canceller on a digital telecommunications network

A method and apparatus for disabling an echo canceller in a framed digital network from a remote terminal that is copuled to the network through an unframed data link. Control is effected by sending N different bit sequences from the remote terminal, where N is the number of different framing alignments that may be imposed on the unframed data by the network. The different sequences are chosen so that one will be framed by the network as the requisite control tone regardless of which framing alignment is actually imposed.

Schroeder, George L.

Monitoring Astronaut Health with DNA Sequencing

In recent years microbe a plethora of microbe populations have been identified onboard the ISS (International Space Station). Approaches for real-time tracking of microbes for routine housekeeping and food/water safety monitoring will be critical for mission safety and crew health on future longer duration missions to the Moon or Mars. This work is a proof-of-concept study demonstrating an end-to-end phylogenetic identification and full genome sequencing effort of multiple microbial populations. Our methodology utilized the ISS flight-certified WetLab-2 molecular toolbox and the Biomolecule Sequencer projects for real-time end-to-end on-orbit microbial biological samples processing and molecular analysis with real time results generated utilizing only field "offline" analytic software. For this experiment we colony-cultured several ISS isolated microorganisms before generation of the pre-sequencing library via the automated VolTRAX device which enabled high library turnover with little wet-bench activity or potential future costly astronaut time. The pre-sequencing library is diluted in loading buffer and injected into the MinION sample port, drawn into the nanopore window by capillary action, and sequenced using the MinKnown. 16S and full genome alignment, nucleotide matching, gene identification, and phylogenetic sorting was accomplished utilizing the Epi2me software and the offline NCBI Blast viral, microbiome, and human somatic databases. In short, the methodologies developed herein replace the myriad of specific, often highly targeted microbiological tests used in the clinical laboratory, which would be difficult if not impossible to currently implement aboard the ISS or in deep space, with a single metagenomics test.

genomics

Impact basins in Southern Daedalia, Mars: Evidence for clustered impactors?

The distribution of ancient massifs and old cratered terrain in the southern Daedalia region indicate the presence of at least two and probably three impact basins of large size. One of these is located near where Craddock et al. placed their center for the Daedalia Basin, but it has very different ring diameters. These basins have rings exceeding 1000 km diameter and overlap significantly with centers separated by 500 to 600 km at nearly identical latitudes of -26 to -29 deg. The smaller westernmost basin appears slightly better preserved, but there is little evidence for obvious superposition that might imply a temporal sequence. Recognizing the improbability of random impacts producing aligned, nearly contemporaneous features, we suggest these basins may have resulted from clustered impactors.

Frey, Herbert

Towards Automating Spacecraft Attitude Sensor Calibration

With a view towards reducing cost and complexity for spacecraft early mission support at the NASA Goddard Space Flight Center (GSFC), efforts are being made to automate the attitude sensor calibration process. This paper addresses one of the major components needed by such a system. The beneficiaries of an improved calibration process are missions that demand moderate to high precision attitude knowledge or that need to perform accurate attitude slews. Improved slew accuracy reduces the time needed for re-acquisition of fine-pointing after each attitude maneuver, Rapid target acquisition can be very important for astronomical targeting or for off-nadir surface feature targeting by Earth-oriented spacecraft. The normal sequence of on-orbit calibration starts with alignment calibration of the star trackers and possibly the Sun sensor. Their relative alignment needs to be determined using a sufficiently large data set so their fields of view are adequately sampled. Next, the inertial reference unit (IRU) is calibrated for corrections to its alignment and scale factors. The IRU biases are estimated continuously by the onboard attitude control system, but the IRU alignment and scale factors are usually determined on the ground using a batch-processing method on a data set that includes several slews sufficient to give full observability of all the IRU calibration parameters. Finally, magnetometer biases, alignment, and its coupling to the magnetic torquers are determined in order io improve momentum management and occasionally for use in the attitude determination system. The detailed approach used for automating calibrations will depend on whether the automated system resides on the ground or on the spacecraft with an ultimate goal of autonomous calibration. Current efforts focus on a ground-based system driving subsystems that could run either on the ground or onboard. The distinction is that onboard calibration should process the data sequentially rather than in a single large batch since onboard computer data storage is limited. Very good batch- processing calibration utilities have been developed and used extensively at NASA/GSFC for mission support but no sequential calibration utilities are available. To meet this need, this paper presents the mathematical description of a sequential IRU calibration system. The system has been tested using flight data from the Rossi X-ray Timing Explorer (RXTE) during a series of attitude slews. The paper also discusses the current state of the overall automated system and describes plans for adding sequential alignment calibration and other additions that will reduce the amount of analyst time and input.

Sedlak, Joseph

Using Fractional Clock-Period Delays in Telemetry Arraying

A set of special digital all-pass finite-impulse- response (FIR) filters produces phase shifts equivalent to delays that equal fractions of the sampling or clock period of a telemetry-data-processing system. These filters have been used to enhance the arraying of telemetry signals that have been received at multiple ground stations from spacecraft (see figure). Somewhat more specifically, these filters have been used to align, in the time domain, the telemetry-data sequences received by the various antennas, in order to maximize the signal-to-noise ratio of the composite telemetric signal obtained by summing the signals received by the antennas. The term arraying in this context denotes a method of enhanced reception of telemetry signals in which several antennas are used to track a single spacecraft. Each antenna receives a signal that comprises a sum of telemetry data plus noise, and these sum data are sent to an arraying combiner for processing. Correlation is the means used to align the set of data from one antenna with that from another antenna. After the data from all the antennas have been aligned in the time domain, they are all added together, sample by sample.

Fort, David

Rapid Test Bed Concentrator (TBC) Alignment Techniques

A new, labor and cost saving method was developed to eliminate the procedure of covering all (220) mirrors and uncovering then one-by-one in sequence to adjust each to the focal plane. This latest method being used to align mirrors of a parabolic solar concentrator utilizes a computer-derived target of discreet images made up of individual mirror reflections on a plane in front of the intended, nominal, focal point. Incorporating this computer technique increases accuracy and gives potential to develop flux distributions required by different receiver designs.

Argoud, M. J.

Real-Time X-ray Radiography Diagnostics of Components in Solid Rocket Motors

Solid rocket motors (SRMs) typically use nozzle materials which are required to maintain their shape as well as insulate the underlying support structure during the motor operation. In addition, SRMs need internal insulation materials to protect the motor case from the harsh environment resulting from the combustion of solid propellant. In the nozzle, typical materials consist of high density graphite, carbon-carbon composites and carbon phenolic composites. Internal insulation of the motor cases is typically a composite material with carbon, asbestos, Kevlar, or silica fibers in an ablative matrix such as EPDM or NBR. For both nozzle and internal insulation materials, the charring process occurs when the hot combustion products heat the material intensely. The pyrolysis of the matrix material takes away a portion of the thermal energy near the wall surface and leaves behind a char layer. The fiber reinforcement retains the porous char layer which provides continued thermal protection from the hot combustion products. It is of great interest to characterize both the total erosion rates of the material and the char layer thickness. By better understanding of the erosion process for a particular ablative material in a specific flow environment, the required insulation material thickness can be properly selected. The recession rates of internal insulation and nozzle materials of SRMs are typically determined by testing in some sort of simulated environment; either arc-jet testing, flame torch testing, or subscale SRMs of different size. Material recession rates are deduced by comparison of pre- and post-test measurements and then averaging over the duration of the test. However, these averaging techniques cannot be used to determine the instantaneous recession rates of the material. Knowledge of the variation in recession rates in response to the instantaneous flow conditions during the motor operation is of great importance. For example, in many SRM configurations the recession of the solid propellant grain can drastically alter the flow-field and effect the recession of internal insulation and nozzle materials. Simultaneous measurement of the overall erosion rate, the development of the char layer, and the recession of the char-virgin interface during the motor operation can be rather difficult. While invasive techniques have been used with limited success, they have serious drawbacks. Break wires or make wire sensors can be installed into a sufficient number of locations in the charring material from which a time history of the charring surface can be deduced. These sensors fundamentally alter the local structure of the material in which they are imbedded. Also, the location of these sensors within the material is not known precisely without the use of an X-ray. To determine instantaneous recession rates, real-time X-ray radiography (X-ray RTR) has been utilized in several SRM experiments at PSU. The X-ray RTR system discussed in this paper consists of an X-ray source, X-ray image intensifier, and CCD camera connected to a capture computer. The system has been used to examine the ablation process of internal insulation as well as nozzle material erosion in a subscale SRM. The X-ray source is rated to 320 kV at 10 mA and has both a large (5.5 mm) and small (3.0 mm) focal spot. The lead-lined cesium iodide X-ray image intensifier produces an image which is captured by a CCD camera with a 1,000 x 1,000 pixel resolution. To produce accurate imagery of the object of interest, the alignment of the X-ray source to the X-ray image intensifier is crucial. The image sequences captured during the operation of an SRM are then processed to enhance the quality of the images. This procedure allows for computer software to extract data on the total erosion rate and the char layer thickness. Figure 1 Error! Reference source not found.shows a sequence of images captured during the operation the subscale SRM with the X-ray RTR system. The X-rayTR system, alignment procedure, uncertainty determination, and image analysis process will be discussed in detail in the full manuscript.

Cortopassi, A. C.

Planar multijunction high voltage solar cell chip

A new innovative planar multijunction solar cell chip for concentrated sunlight applications is proposed. The chip consists of many voltage-generating regions, called unit cells, which are connected in series within a single silicon wafer, thereby providing a high open-circuit voltage at multiple sun illumination levels. The unit cells are fabricated on 75 micron thick p-type single crystal silicon substrate. Each chip consists of 1.42 x 9.63 mm n(+)/p collecting junctions on the back of the wafer, while the illuminated front surface area is divided into 0.3 micron deep n(+) regions. The fabrication sequence includes standard degreasing and cleaning procedures, double-sided alignment photomasking, introduction of boron and phosphorus impurities, and photolithography. The open circuit voltage of the chip increased rapidly with illumination up to about 4 AM1 suns, and then began to saturate at the sum of the individual unit cell voltages of 3.5 above 4 AM1 suns. A short circuit density per unit cell of 300 mA/sq cm at 20 AM1 suns was observed.

Valco, G. J.

Halogen occultation experiment (HALOE) performance verification test procedure

The Performance Verification Test Procedure is given for the Halogen Occultation Experiment (HALOE) instrument, which is being developed in house at the Langley Research Center for the Upper Atmosphere Research Satellite (UARS). This procedure is used for comprehensive performance testing of the HALOE instrument which occurs before, during, and after flight environmental tests. The radiometric performance tests include noise, drift, linearity, instantaneous field-of-view, cal wheel gas cell characterization, and self thermal emissions. Pointer/tracker performance tests include sun sensor performance, gimbal performance, control system performance, and boresight alignment. In addition, the instrument is tested functionally in simulated orbit sequences and all command operating modes are exercised. The data analysis required for each test is specified and pass/fail criteria are given where applicable. This test will fully demonstrate the HALOE instrument's ability to achieve science mission requirements. The HALOE instrument is a gas correlation radiometer that measures vertical distribution of eight upper atmospheric constituents: O3, HCl, HF, NO, CH4, H2O, NO2, and CO2.

Mauldin, L. E., III

Isolation of new ribozymes from a large pool of random sequences

An iterative in vitro selection procedure was used to isolate a new class of catalytic RNAs (ribozymes) from a large pool of random-sequence RNA molecules. These ribozymes ligate two RNA molecules that are aligned on a template by catalyzing the attack of a 3'-hydroxyl on an adjacent 5'-triphosphate - a reaction similar to that employed by the familiar protein enzymes that synthesize RNA. The corresponding uncatalyzed reaction also yields a 3',5'-phosphodiester bond. In vitro evolution of the population of new ribozymes led to improvement of the average ligation activity and the emergence of ribozymes with reaction rates 7 million times faster than the uncatalyzed reaction rate.

Bartel, David P.

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.

Detailed Test Plan Redundant Sensor Strapdown IMU Evaluation Program

The test plan for a redundant sensor strapdown inertial measuring unit evaluation program is presented. The subjects discussed are: (1) test philosophy and limitations, (2) test sequence, (3) equipment specifications, (4) general operating procedures, (5) calibration procedures, (6) alignment test phase, and (7) navigation test phase. The data and analysis requirements are analyzed.

Hartwell, T.

STRAP V - Higher accuracy, lower drift attitude control system

The STRAP V system was developed to provide higher accuracy and lower limit cycle fine pointing (+ or - 7 arcseconds) in all three axes at targets which cannot be tracked by startrackers or solar trackers. The system provides an increase in pointing performance over that obtainable with the STRAP IV (1) Attitude Control System (ACS). The STRAP IV concept of third axis updates is utilized to reduce pointing errors, using the flight-proven STRAP III (2) system as a first stage. Flight aspect photographs and telemetry records show that the STRAP V objectives have been met. The STRAP IV major error contributors have been significantly reduced and the tracking flexibility has been increased with only minor error contributions. Attention is given to the basic STRAP III control modes, major STRAP IV system error sources, tuned restrained inertial gyros (TRIGs), the programmable sequence timer, the STRAP V control box, third axis update, system gyro alignments, and STRAP V operational capabilities.

Budney, T. J.

NASA Tech Briefs, May 2006

T-Shaped Emitter Metal Structures for HBTs Rigorous Estimation of SNR of a PSK Communication Link Advanced Ka-Band Transceiver With Monopulse Tracking EMI Filters for Low-Temperature Applications Lightweight Electronic Camera for Research on Clouds Pilot Weather Advisor System Waveguide Power-Amplifier Module for 80 to 150 GHz Better Back Contacts for Solar Cells on Flexible Substrates Topics covered include:Tunable, Highly Stable Lasers for Coherent Lidar; Improved Photon-Emission-Microscope System; Program Synthesizes UML Sequence Diagrams; Aspect-Oriented Subprogram Synthesizes UML Sequence Diagrams; Updated Computational Model of Cosmic Rays Near Earth; Software for Alignment of Segments of a Telescope Mirror; Simulation of Dropping of Cargo With Parachutes; DAVE-ML Utility Program; Robust Control for the Mercury Laser Altimeter; Thermally Stable Piezoelectric and Pyroelectric Polymers; Combustion Synthesis of Ca3(PO4)2 Net-Shape Surgical Implants; Stochastic Representation of Chaos Using Terminal Attractors; Two High-Temperature Foil Journal Bearings; Using Plates To Represent Fillets in Finite-Element Modeling; Repairing Chipped Silicide Coatings on Refractory Metal Substrates; Simplified Fabrication of Helical Copper Antennas; Graded-Index "Whispering-Gallery" Optical Microresonators; Optical Profilometers Using Adaptive Signal Processing; Manufacture of Sparse-Spectrum Optical Microresonators; Exact Tuning of High-Q Optical Microresonators by Use of UV; Automation for "Direct-to" Clearances in Air-Traffic Control; Improved Traps for Removing Gases From Coolant Liquids; and Lunar Constellation of Frozen Elliptical Inclined Orbits.

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