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NASA NTRS1991

Exploration of RNA structure spaces

In order to understand the structure of real structure spaces, we are studying the 5S rRNA structure space experimentally. A plasmid containing a synthetic 5S rRNA gene, two rRNA promoters, and transcription terminators has been assembled. Assays are conducted to determine if the foreign 5S rRNA is expressed, and to see whether or not it is incorporated into ribosomes. Evolutionary competition is used to determine the relative fitness of strains containing the foreign 5S rRNA and a control 5S rRNA. By using site directed mutagenesis, a number of mutants can be made in order to study the boundaries of the structure space and how sharply defined they are. By making similar studies in the vicinity of structure space, it will be possible to determine how homogeneous the 5S rRNA structure space is. Useable experimental protocols have been developed, and a number of mutants have already been studied. Initial results suggest an explanation of why single stranded regions of the RNA are less subject to mutation than double stranded regions.

Fox, G. E.↗
NASA NTRS2017

The Universal Space Transponder: A Next Generation Software Defined Radio

The Universal Space Transponder (UST) is a next generation transponder developed at the Jet Propulsion Laboratory to meet a large variety of telecom, navigation, and radio science needs for future deep-space and near-Earth missions. This paper details the UST software defined radio design and describes how the combination of a modular hardware architecture and in-flight reprogrammability enables a new level of flexibility and expandability for a space transponder. The UST uses common power and digital processing assemblies that can be integrated with a variety of RF modules and is capable of simultaneous, multiband operations with data rates up to 37.5 Mbps RX and 300 Mbps TX. This allows a single radio to support all the direct-to-Earth and relay communication requirements for even complex mission scenarios, reducing the total cost, mass, and power. The discussion includes a description of the current UST engineering models that have been built and tested, as well as details about the next generation capabilities supported by UST, including advanced link coding and modulation, radiometric techniques, and in-radio protocol handling. Details are also presented on RF modules and digital processing in development for radio science and astronomy purposes, including a bistatic radar receiver and broadband planetary emissions receiver. These will demonstrate the ability to integrate low-cost science instruments into the UST architecture, further expanding the versatility of the UST.

Jedrey, Thomas↗
NASA NTRS2010

Fine-Water-Mist Multiple-Orientation-Discharge Fire Extinguisher

A fine-water-mist fire-suppression device has been designed so that it can be discharged uniformly in any orientation via a high-pressure gas propellant. Standard fire extinguishers used while slightly tilted or on their side will not discharge all of their contents. Thanks to the new design, this extinguisher can be used in multiple environments such as aboard low-gravity spacecraft, airplanes, and aboard vehicles that may become overturned prior to or during a fire emergency. Research in recent years has shown that fine water mist can be an effective alternative to Halons now banned from manufacture. Currently, NASA uses carbon dioxide for fire suppression on the International Space Station (ISS) and Halon chemical extinguishers on the space shuttle. While each of these agents is effective, they have drawbacks. The toxicity of carbon dioxide requires that the crew don breathing apparatus when the extinguishers are deployed on the ISS, and Halon use in future spacecraft has been eliminated because of international protocols on substances that destroy atmospheric ozone. A major advantage to the new system on occupied spacecraft is that the discharged system is locally rechargeable. Since the only fluids used are water and nitrogen, the system can be recharged from stores of both carried aboard the ISS or spacecraft. The only support requirement would be a pump to fill the water and a compressor to pressurize the nitrogen propellant gas. This system uses a gaseous agent to pressurize the storage container as well as to assist in the generation of the fine water mist. The portable fire extinguisher hardware works like a standard fire extinguisher with a single storage container for the agents (water and nitrogen), a control valve assembly for manual actuation, and a discharge nozzle. The design implemented in the proof-of-concept experiment successfully extinguished both open fires and fires in baffled enclosures.

Butz, James R.↗
NASA NTRSDate not supplied

Firmware Architecture of the ARMADAS Bolting Robot

The Automated Reconfigurable Mission Adaptive Digital Assembly Systems (ARMADAS) project, under development at NASA Ames Research Center, has demonstrated on-ground autonomous robotic assembly of extensive digital structures, and it is now moving forward towards in-space demonstration. The ARMADAS system comprises of the operation software, the operation user interface (opsUI), and a swarm of robots. The robotic system consists of a multitude of collaborative agents specifically designed to transport, place and bolt the building blocks, called voxels (volumetric pixels). This paper focuses on the bolting robot, referred to as Mobile Metamaterial Internal Co-Integrator (MMIC-I). MMIC-I is a battery-powered crawling robot. It navigates the structure through extension, contraction and gripping. Two distinct controller boards operate the robot's two symmetric modules, referred to as module A and B. Board A is the master board: it coordinates motion planning and motion primitives execution, hosts the WiFi client, performs periodic self-assessment and system idle check and triggers faults if anomalies are detected. Board B periodically sends a heartbeat to board A, through a wired communication channel that uses the Serial protocol. Additionally, board A's WiFi client receives heartbeat packet requests or motion/bolting commands from a dedicated server board, and acknowledges reception sending back a response heartbeat packet containing information about the overall robot status, e. g. electrical current and voltage values, target and actual angles, operating mode, fault status. Whenever a motion command is sent, the motion planning section of the firmware determines the current robot configuration, using Inertial Measurement Unit readings and the motors Pulse Width Modulation values. Afterwards, it calculates the list of primitives needed to reach the target state, and controls their execution in the proper order. MMIC-I can receive and execute motion and bolting commands only when it is in operational mode. MMIC-I has three operating modes: standby, operational and safed. Standby mode is automatically entered upon startup. While in standby mode, all motors are powered off, and the only accepted commands are the ones relative to a change of mode and heartbeat packet request. Fault detection causes the robot to automatically enter safed or standby mode. Whenever the detected fault occurs within a motion and requires immediate intervention, e. g. an over-current situation, the robot enters safed mode. Safed mode powers off all motors except for the locomotion module, thus preventing the robot from collapsing. Conversely, when the detected fault doesn't require immediate intervention (low battery warning, for instance), the robot enters standby mode after completing the ongoing motion. This paper provides a detailed discussion of MMIC-I's firmware architecture. It accurately describes the implementation approach for each module: sensor data reading, motor control and actuation, WiFi server-client communication, intra-boards Serial communication, operating modes and autonomous fault detection, motion planning, coordination and execution, etc. Moreover, in support of the software description, this paper includes a thorough characterization of MMIC-I's hardware and avionics.

In-space assembly↗
NASA NTRS1992

The VLBA correlator: Real-time in the distributed era

The correlator is the signal processing engine of the Very Long Baseline Array (VLBA). Radio signals are recorded on special wideband (128 Mb/s) digital recorders at the 10 telescopes, with sampling times controlled by hydrogen maser clocks. The magnetic tapes are shipped to the Array Operations Center in Socorro, New Mexico, where they are played back simultaneously into the correlator. Real-time software and firmware controls the playback drives to achieve synchronization, compute models of the wavefront delay, control the numerous modules of the correlator, and record FITS files of the fringe visibilities at the back-end of the correlator. In addition to the more than 3000 custom VLSI chips which handle the massive data flow of the signal processing, the correlator contains a total of more than 100 programmable computers, 8-, 16- and 32-bit CPUs. Code is downloaded into front-end CPU's dependent on operating mode. Low-level code is assembly language, high-level code is C running under a RT OS. We use VxWorks on Motorola MVME147 CPU's. Code development is on a complex of SPARC workstations connected to the RT CPU's by Ethernet. The overall management of the correlation process is dependent on a database management system. We use Ingres running on a Sparcstation-2. We transfer logging information from the database of the VLBA Monitor and Control System to our database using Ingres/NET. Job scripts are computed and are transferred to the real-time computers using NFS, and correlation job execution logs and status flow back by the route. Operator status and control displays use windows on workstations, interfaced to the real-time processes by network protocols. The extensive network protocol support provided by VxWorks is invaluable. The VLBA Correlator's dependence on network protocols is an example of the radical transformation of the real-time world over the past five years. Real-time is becoming more like conventional computing. Paradoxically, 'conventional' computing is also adopting practices from the real-time world: semaphores, shared memory, light-weight threads, and concurrency. This appears to be a convergence of thinking.

Wells, D. C.↗
NASA NTRS1990

Feasibility study, software design, layout and simulation of a two-dimensional fast Fourier transform machine for use in optical array interferometry

The NASA-Cornell Univ.-Worcester Polytechnic Institute Fast Fourier Transform (FFT) chip based on the architecture of the systolic FFT computation as presented by Boriakoff is implemented into an operating device design. The kernel of the system, a systolic inner product floating point processor, was designed to be assembled into a systolic network that would take incoming data streams in pipeline fashion and provide an FFT output at the same rate, word by word. It was thoroughly simulated for proper operation, and it has passed a comprehensive set of tests showing no operational errors. The black box specifications of the chip, which conform to the initial requirements of the design as specified by NASA, are given. The five subcells are described and their high level function description, logic diagrams, and simulation results are presented. Some modification of the Read Only Memory (ROM) design were made, since some errors were found in it. Because a four stage pipeline structure was used, simulating such a structure is more difficult than an ordinary structure. Simulation methods are discussed. Chip signal protocols and chip pinout are explained.

Boriakoff, Valentin↗
NASA NTRS2001

Bio-Optical Measurement and Modeling of the California Current and Polar Oceans

The principal goals of our research are to validate standard or experimental products through detailed bio-optical and biogeochemical measurements, and to combine ocean optical observations with advanced radiative transfer modeling to contribute to satellite vicarious radiometric calibration and advanced algorithm development. To achieve our goals requires continued efforts to execute complex field programs globally, as well as development of advanced ocean optical measurement protocols. We completed a comprehensive set of ocean optical observations in the California Current, Southern Ocean, Indian Ocean requiring a large commitment to instrument calibration, measurement protocols, data processing and data merger. We augmented separately funded projects of our own, as well as others, to acquire ill situ data sets we have collected on various global cruises supported by separate grants or contracts. In collaboration with major oceanographic ship-based observation programs funded by various agencies (CalCOFI, US JGOFS, NOAA AMLR, INDOEX and Japan/East Sea) our SIMBIOS effort has resulted in data from diverse bio-optical provinces. For these global deployments we generate a high-quality, methodologically consistent, data set encompassing a wide-range of oceanic conditions. Global data collected in recent years have been integrated with our on-going CalCOFI database and have been used to evaluate SeaWiFS algorithms and to carry out validation studies. The combined database we have assembled now comprises more than 700 stations and includes observations for the clearest oligotrophic waters, highly eutrophic blooms, red-tides and coastal case 2 conditions. The data has been used to validate water-leaving radiance estimated with SeaWiFS as well as bio-optical algorithms for chlorophyll pigments. The comprehensive data is utilized for development of experimental algorithms (e.g. high-low latitude pigment transition, phytoplankton absorption, and cDOM). During this period we completed 9 peer-reviewed publications in high quality journals, and presented aspects of our work at more than 10 scientific conferences.

Mitchell, B. Greg↗
NASA NTRS2002

The SeaWiFS Bio-Optical Archive and Storage System (SeaBASS): Current Architecture and Implementation

Satellite ocean color missions require an abundance of high-quality in situ measurements for bio-optical and atmospheric algorithm development and post-launch product validation and sensor calibration. To facilitate the assembly of a global data set, the NASA Sea-viewing Wide Field-of-view (SeaWiFS) Project developed the Seafaring Bio-optical Archive and Storage System (SeaBASS), a local repository for in situ data regularly used in their scientific analyses. The system has since been expanded to contain data sets collected by the NASA Sensor Intercalibration and Merger for Biological and Interdisciplinary Oceanic Studies (SIMBIOS) Project, as part of NASA Research Announcements NRA-96-MTPE-04 and NRA-99-OES-99. SeaBASS is a well moderated and documented hive for bio-optical data with a simple, secure mechanism for locating and extracting data based on user inputs. Its holdings are available to the general public with the exception of the most recently collected data sets. Extensive quality assurance protocols, comprehensive data and system documentation, and the continuation of an archive and relational database management system (RDBMS) suitable for bio-optical data all contribute to the continued success of SeaBASS. This document provides an overview of the current operational SeaBASS system.

Werdell, P. Jeremy↗
NASA NTRSDate not supplied

rHealth One Demonstration Aboard ISS: A Microfluidic Bioanalyzer Based on Sheath/Hydrodynamic Focusing Flow Cytometry

The Exploration Medical Capability (ExMC) element aims to provide astronauts with the means for their own health monitoring, diagnosis, and treatment during exploration missions. As space flight ventures further from earth, the need for autonomous medical care increases under greater constraints on size, mass, and resources. One pillar for diagnosis that crew would be separated from is laboratory analysis. Even now on ISS, blood samples must be collected and returned to earth for testing. In response, ExMC is assessing how assays for hematology, bone health, radiation exposure etc. could be addressed through commercial-off-the-shelf (COTS) and Small Business Innovation Research (SBIR) funded bioanalyzers that are miniaturizing lab technology. Since missions will reach distances where there are no timely replacements, validation on the International Space Station (ISS) is a necessary part of that assessment. In partnership with NASA Johnson Space Center (JSC) Immunology Lab and the Research Operations and Integration (ROI) element, ExMC conducted a technology demonstration on ISS of the rHEALTH ONE, a flow cytometry based bioanalyzer, to assess future devices based on this design. In flow cytometry, there are predominantly three ways to focus the cells (or particles) into a single file stream: hydrodynamic focusing, microcapillary, and acoustic focusing. The rHEALTH ONE utilizes the commercial standard, sheath-based hydrodynamic focusing. rHEALTH itself represents both a company and a suite of medical tools NASA has funded through SBIR grants towards the development of a promising diagnosis instrument for exploration missions. rHEALTH ONE is the interim version of the technology, functional as a benchtop analyzer and test bed for the next generation of rHEALTH in development. Several modifications were made to the rHEALTH ONE analyzer for operation in microgravity. For function, fluid management was key. A sheath-based analyzer uses sheath fluid to flow the sample, cleaning fluid to prevent biological contamination, and a reservoir to collect the liquid waste. The rHEALTH ONE analyzer uses bottles dependent on gravity to separate the air and liquid pathways and keep the liquids contained. It uses only 1 psig of air pressure to directly push the liquid through the device, requiring little-to-no resistance at inlet and outlet. A microtubing assembly with self-sealing luer connectors was designed – featuring 0.014 mm thick durable medical balloons to hold water inside the supply bottles – to create safe containment and easy access for the crew while maintaining the analyzer’s function. For safety, copper tape was added to the interior of the plastic housing to reduce electromagnetic interference, fluid and electrical connections were secured against vibration and leaks, and gaps were further sealed to ensure containment of the optical block and lasers. Water as the sheath and cleaning fluid and TOX 0 samples were used to reduce the biohazard risk to the crew. In May 2022, European Space Agency astronaut Samantha Cristoforetti demonstrated the rHEALTH ONE aboard ISS for its sample loading, flow cytometry, and data collection capability in microgravity. The JSC Immunology Lab provided flow cytometry expertise, designed the sample test protocol, and manufactured and benchmarked the flight samples on a ‘gold-standard’ flow cytometer. The analyzer was primed with water, purged of air, and four calibration solutions of polystyrene microparticles were tested to characterize its performance. Tightly controlled procedures and excellent execution prevented air bubble interference during air/water separation, fluid transfer, sample mixing and loading. Data showed a slight increase in signal noise on 3 of 5 channels and an anomaly of fluorophores migrating in one sample (confirmed by JSC post-flight). Flight results correctly detected the change in sample, matched the analyzer’s ground performance, and were consistent with the gold-standard’s ground results. Although areas were noted for improvement, these outcomes signified complete mission success.

R. S. Miller↗
NASA NTRS2001

Combining Information on Multiple Detection Techniques to Estimate the Effect of Patent Foramen Ovale on Susceptibility to Decompression Illness

The assembly and the maintenance of the International Space Station is expected to require hundreds of extravehicular excursions (EVA's) in the next 10 years. During an EVA, in order to allow movement and bending of limbs, spacesuit pressures are reduced to about 4.3 psi. as compared with about 14.7 psi. for normal atmospheric pressure at sea level. However, the exposure of astronauts to reduced pressures in spacesuits, is conducive to fonnation and growth of gas bubbles within venous blood or tissues, which could cause decompression illness (DCI), a pathology best known to occur among deep-sea divers when they return to the surface. To reduce the risk of DCI, astronauts adjust to the reduced pressure in stages for a prolonged time known as a "pre-breathe" period prior to their extravehicular activity. Despite the use of pre-breathe protocols, an increased risk of DCI can arise for about 25% of humans who have a small hole, known as a patent foramen ovale (PFO), between two chambers of the heart. The atrial septum's fossa oval is, an embryological remnant of a flap between the septae primum and secundum allows fetal right atrial blood to pass into the left atrium, and usually closes after birth (Hagen, et al,. 1984). If fusion does not occur, a valve-like opening, the foramen ovale persists between the two atria. It has been suggested that astronauts with PFO's might be at greater risk of stroke or other serious neurological DCI because bubbles from a venous site may traverse a PFO, travel to the aorta and then enter the cerebral circulatory system causing a stroke (Figure 1). Astronauts are not now screened for PFO's, however consideration is being given to doing so. Here, we study three main methods abbreviated here as "ITE", "TCD" and "TEE", for detecting PFO's in living subjects. All involve the introduction of bubbles into a vein, immediately after which a sensory probe attempts to detect the bubbles in systemic circulation. Presence of the injected bubbles in the systemic circulation is indicative of a PFO. More detailed descriptions are given after the explanation of PFO's under Figure I. Even if a true PFO affects the risk of DCI, there remains a question of how effective screening would be if the detection method has errors of omission and/or commission. Of the three methods studied here, TEE is the "gold standard", matching autopsy results with near-perfect sensitivity and specificity (Schneider, et al. , 1996). However TEE is also the most difficult method to implement, requiring an internal esophagal probe, and is therefore not widely used. Currently, the easiest to use and most common PFO detection method is TTE, which uses an external chest probe. This method has a specificity of near 100%, but suffers from a low sensitivity rate (about 30%). More recently, TCD has been developed, which uses ultrasound probes to detect the presence of bubbles in cerebral arteries. Studies indicate that TCD is quite effective, having a sensitivity of about 91% and a specificity of about 93% (Droste, et al., 1999) when applied correctly, however implementation is difficult and requires considerable training.

Feiveson, Alan H.↗
NASA NTRS1993

Making tomorrow's mistakes today: Evolutionary prototyping for risk reduction and shorter development time

In the early days of JPL's solar system exploration, each spacecraft mission required its own dedicated data system with all software applications written in the mainframe's native assembly language. Although these early telemetry processing systems were a triumph of engineering in their day, since that time the computer industry has advanced to the point where it is now advantageous to replace these systems with more modern technology. The Space Flight Operations Center (SFOC) Prototype group was established in 1985 as a workstation and software laboratory. The charter of the lab was to determine if it was possible to construct a multimission telemetry processing system using commercial, off-the-shelf computers that communicated via networks. The staff of the lab mirrored that of a typical skunk works operation -- a small, multi-disciplinary team with a great deal of autonomy that could get complex tasks done quickly. In an effort to determine which approaches would be useful, the prototype group experimented with all types of operating systems, inter-process communication mechanisms, network protocols, packet size parameters. Out of that pioneering work came the confidence that a multi-mission telemetry processing system could be built using high-level languages running in a heterogeneous, networked workstation environment. Experience revealed that the operating systems on all nodes should be similar (i.e., all VMS or all PC-DOS or all UNIX), and that a unique Data Transport Subsystem tool needed to be built to address the incompatibilities of network standards, byte ordering, and socket buffering. The advantages of building a telemetry processing system based on emerging industry standards were numerous: by employing these standards, we would no longer be locked into a single vendor. When new technology came to market which offered ten times the performance at one eighth the cost, it would be possible to attach the new machine to the network, re-compile the application code, and run. In addition, we would no longer be plagued with lack of manufacturer support when we encountered obscure bugs. And maybe, hopefully, the eternal elusive goal of software portability across different vendors' platforms would finally be available. Some highlights of our prototyping efforts are described.

Friedman, Gary↗
NASA NTRS2016

Essential SpaceWire Hardware Capabilities for a Robust Network

The Geostationary Operational Environmental Satellite R-Series Program (GOES-R) mission is a joint program between National Oceanic & Atmospheric Administration (NOAA) and National Aeronautics & Space Administration (NASA) Goddard Space Flight Center (GSFC). GOES-R project selected SpaceWire as the best solution to satisfy the desire for simple and flexible instrument to spacecraft command and telemetry communications. GOES-R development and integration is complete and the observatory is scheduled for launch October 2016. The spacecraft design was required to support redundant SpaceWire links for each instrument side, as well as to route the fewest number of connections through a Slip Ring Assembly necessary to support Solar pointing instruments. The final design utilized two different router designs. The SpaceWire standard alone does not ensure the most practical or reliable network. On GOES-R a few key hardware capabilities were identified that merit serious consideration for future designs. Primarily these capabilities address persistent port stalls and the prevention of receive buffer overflows. Workarounds were necessary to overcome shortcomings that could be avoided in future designs if they utilize the capabilities, discussed in this paper, above and beyond the requirements of the SpaceWire standard.

SpaceWire↗
NASA NTRS1999

Induction of three-dimensional assembly of human liver cells by simulated microgravity

The establishment of long-term cultures of functional primary human liver cells (PHLC) is formidable. Developed at NASA, the Rotary Cell Culture System (RCCS) allows the creation of the unique microgravity environment of low shear force, high-mass transfer, and 3-dimensional cell culture of dissimilar cell types. The aim of our study was to establish long-term hepatocyte cultures in simulated microgravity. PHLC were harvested from human livers by collagenase perfusion and were cultured in RCCS. PHLC aggregates were readily formed and increased up to 1 cm long. The expansion of PHLC in bioreactors was further evaluated with microcarriers and biodegradable scaffolds. While microcarriers were not conducive to formation of spheroids, PHLC cultured with biodegradable scaffolds formed aggregates up to 3 cm long. Analyses of PHLC spheroids revealed tissue-like structures composed of hepatocytes, biliary epithelial cells, and/or progenitor liver cells that were arranged as bile duct-like structures along nascent vascular sprouts. Electron microscopy revealed groups of cohesive hepatocytes surrounded by complex stromal structures and reticulin fibers, bile canaliculi with multiple microvilli, and tight cellular junctions. Albumin mRNA was expressed throughout the 60-d culture. A simulated microgravity environment is conducive to maintaining long-term cultures of functional hepatocytes. This model system will assist in developing improved protocols for autologous hepatocyte transplantation, gene therapy, and liver assist devices, and facilitate studies of liver regeneration and cell-to-cell interactions that occur in vivo.

NASA Center JSC↗
NASA NTRSDate not supplied

Performance Evaluation of a Cartesian Move Algorithm for the LSMS Family of Cable-Driven Cranes

The Lightweight Surface Manipulation System, or LSMS, is a family of cable-actuated cranes, designed for surface operations on the Moon and Mars. Its structural design focuses on three fundamental traits for space environments: a high payload-to-mass ratio to reduce launch costs, compactness for space travel in reduced cargo bays, and self-erecting capabilities for operations before astronauts arrive. The resulting design confers nonlinear and switched dynamics to the LSMS, making these cranes an interesting challenge for autonomous and teleoperated applications. One of the protocols required for these applications is a Cartesian move algorithm. This algorithm is responsible for placing the end effector of the crane at a desired location, specified in Cartesian coordinates. This paper evaluates the performance of a Cartesian move algorithm developed for the LSMS at the NASA Langley Research Center. To this end, several payload manipulation tasks were performed with LSMS-L35, the smallest version within the LSMS family. The evaluated algorithm can place the end effector with accuracy, as shown in the hardware and simulation tests.

cable-actuated↗
NASA NTRS2018

Qualifying Bulk Metallic Glass Gear Materials for Spacecraft Applications

NASA is evaluating bulk metallic glass (BMG) gears for extreme environment (cryogenic) applications; e.g., Europa Lander. The main purpose of this report is to recommend a quality assurance (QA) protocol for the production of consistent and reliable gear castings. Currently, there are two, separate manufacturers involved; Materion Corporation produces re-melt stock and Visser Precision Cast, Inc. produces BMG castings. Division of the existing alloy specifications document into material supplier-specific documents is proposed. Until a composition-based specification is established, processability- and performance-based methodologies may be an option during manufacturing development. It is recommended that each of the documents has a distinct focus; the processing behavior of crystalline feedstock, and the mechanical behavior of amorphous castings. The report addresses the “gray area” in QA testing between these materials suppliers by outlining an equitable division of responsibilities. The use of a number of dedicated reference dies to produce witness materials for physical and mechanical property evaluation is strongly recommended. Allocation of QA tests between screen testing of gear castings, and formal testing of witness materials, including cryogenic properties, is also proposed. The most effective suite of tests for both physical and mechanical property determination are down-selected from available national and international standards. The contents of this report create a solid basis for selection of QA tests, thereby setting the stage for negotiations between the materials suppliers and the customer.This technical paper:a) provides recommendations to advance materials specifications document JPL D-56223,Rev. A from “draft” to “preliminary” status;b) proposes division of the current document into two supplier-specific documents;c) separates testing methods by “ingots/castings” and “witness materials”;d) classifies test methodology based on “processability” and “performance”;e) assembles candidate ASTM specifications for “screen testing” and “formal testing”;f) identifies standard tests compatible with evaluation of BMG flexspline gears;g) suggests division of responsibilities for processability and performance testing; andh) addresses the QA “gray area” between production of ingots and castings.The report recommends that:a) processability- and performance-based QA methods might be used in lieu ofcompositional specifications during manufacturing development only;b) reference dies be designed for “acceptance” and “qualification” castings;c) witness materials provide physical and mechanical property data;d) formal testing be conducted at ambient and cryogenic temperatures; ande) processing/properties be correlated with compositional variations.

Hales, Stephen J.↗
NASA NTRS2007

"Smart" Sensor Module

An assembly that contains a sensor, sensor-signal-conditioning circuitry, a sensor-readout analog-to-digital converter (ADC), data-storage circuitry, and a microprocessor that runs special-purpose software and communicates with one or more external computer(s) has been developed as a prototype of "smart" sensor modules for monitoring the integrity and functionality (the "health") of engineering systems. Although these modules are now being designed specifically for use on rocket-engine test stands, it is anticipated that they could also readily be designed to be incorporated into health-monitoring subsystems of such diverse engineering systems as spacecraft, aircraft, land vehicles, bridges, buildings, power plants, oilrigs, and defense installations. The figure is a simplified block diagram of the "smart" sensor module. The analog sensor readout signal is processed by the ADC, the digital output of which is fed to the microprocessor. By means of a standard RS-232 cable, the microprocessor is connected to a local personal computer (PC), from which software is downloaded into a randomaccess memory in the microprocessor. The local PC is also used to debug the software. Once the software is running, the local PC is disconnected and the module is controlled by, and all output data from the module are collected by, a remote PC via an Ethernet bus. Several smart sensor modules like this one could be connected to the same Ethernet bus and controlled by the single remote PC. The software running in the microprocessor includes driver programs for operation of the sensor, programs that implement self-assessment algorithms, programs that implement protocols for communication with the external computer( s), and programs that implement evolutionary methodologies to enable the module to improve its performance over time. The design of the module and of the health-monitoring system of which it is a part reflects the understanding that the main purpose of a health-monitoring system is to detect damage and, therefore, the health-monitoring system must be able to function effectively in the presence of damage and should be capable of distinguishing between damage to itself and damage to the system being monitored. A major benefit afforded by the self-assessment algorithms is that in the output of the module, the sensor data indicative of the health of the engineering system being monitored are coupled with a confidence factor that quantifies the degree of reliability of the data. Hence, the output includes information on the health of the sensor module itself in addition to information on the health of the engineering system being monitored.

Mahajan, Ajay↗
NASA NTRSDate not supplied

Enumeration and Fluorescence In Situ Hybridization of Microbial Bioburden on Cleanroom Surfaces

Introduction: Microorganisms are everywhere on Earth, even in the cleanest of places. Spacecraft assembly cleanrooms can harbor low levels of living and dead microbial cells (e.g., [1,2]), and cleanroom bioburden can also include organic molecules from industrial sources and in situ biomass. Life detection missions require careful attention to avoid contaminants that can be easily convoluted with analytical targets. We are evaluating epifluorescent microscopy and fluorescence in situ hybridization (FISH) as methods to complement organic contamination detection techniques. Epifluorescent cell counting offers an accurate and cost-effective way to quantify low levels of surface biomass. FISH could allow for the identification of residual organisms, and can be targeted to detect active populations of specific organisms such as bacteria known to resist cleaning procedures. This effort is part of a larger study that is concentrated on characterizing the surface and airborne molecular organic contamination background in Johnson Space Center (JSC) Astromaterials curation laboratories and Goddard Space Flight Center (GSFC) spacecraft assembly rooms, and understanding contaminants in the context of cleaning procedures and residual bioburden. Methods: Samples were collected by swabbing surfaces in ISO 5 and ISO 7 equivalent cleanrooms at JSC. Swabs for FISH were fixed in 4% paraformaldehyde (PFA) for 3 hours and then stored in 1:1 ethanol:PBS, while swabs for cell counting were stored in 4% PFA until analysis to avoid any cell loss during centrifugation that could impact quantification of very low biomass samples. Cell counting was performed with SYBR Gold as in [3], but adapted for very low biomass. FISH was performed as in [4], using DAPI as a counterstain for all DNA-containing cells. Negative controls included wells with no probe applied, to test for natural fluorescence, as well as the nonsense probe NONEUB (reverse complement of EUB338) to evaluate non-specific probe binding. Results and Discussion: Cleanroom surfaces had 102-103 cells cm-2. The extremely low biomass of these samples was challenging for enumeration, and required careful and routine use of “field” and laboratory blanks. FISH was performed with the general archaeal and bacterial probes ARCH915 and EUB338 (EUBMIX, [4]), probe GAMBET ([4]), and PSE227, which targets the genus Pseudomonas [5]). The latter two probes were selected because Pseudomonas spp. and other Gammaproteobacteria have not been isolated from cleanroom surfaces but do appear frequently in rRNA gene libraries from these surfaces. While some active bacteria were identified (Fig. 1c), most cells detectable by DAPI did not have a strong or any fluorescent signal (e.g., Fig. 1d), indicating that the vast majority of cells are dead or inactive. This suggests that cleaning protocols are effective at inactivating microbial contaminants, but that dead or inactive cells can remain on surfaces. Cells were often clumped in a weakly autofluorescent matrix, possibly biofilm material (Fig. 1c,d). We also observed other particulate material that was collected by the swabs, including apparent textile fibers (Fig. 1b). Our results are consistent with other studies that show that the bioburden present in clean rooms includes active, dormant, and dead cells. We will discuss how FISH and epifluorescent cell counting could be applied in planetary protection protocols, including the advantages and disadvantages of FISH and cell counting for routine use, as well as different possible applications for more specialized FISH procedures. References: [1] Moissl-Eichinger et al. (2015) Sci Rep, 5, 9156 [2] Hendrickson et al. (2021) Microbiome, 9, 238 [3] Jones et al. (2017) Appl Environ Microbiol, 83, e00909-17 [4] Jones et al. (2015) Appl Environ Microbiol, 81, 1242-1250. [5] Watt et al. (2006) Environ Microbiol, 8, 871-884

C J Huff↗
NASA NTRS2012

Molecular Technique to Reduce PCR Bias for Deeper Understanding of Microbial Diversity

Current planetary protection policies require that spacecraft targeted to sensitive solar system bodies be assembled and readied for launch in controlled cleanroom environments. A better understanding of the distribution and frequency at which high-risk contaminant microbes are encountered on spacecraft surfaces would significantly aid in assessing the threat of forward contamination. However, despite a growing understanding of the diverse microbial populations present in cleanrooms, less abundant microbial populations are probably not adequately taken into account due to technological limitations. This novel approach encompasses a wide spectrum of microbial species and will represent the true picture of spacecraft cleanroom-associated microbial diversity. All of the current microbial diversity assessment techniques are based on an initial PCR amplification step. However, a number of factors are known to bias PCR amplification and jeopardize the true representation of bacterial diversity. PCR amplification of a minor template appears to be suppressed by the amplification of a more abundant template. It is widely acknowledged among environmental molecular microbiologists that genetic biosignatures identified from an environment only represent the most dominant populations. The technological bottleneck overlooks the presence of the less abundant minority population and may underestimate their role in the ecosystem maintenance. DNA intercalating agents such as propidium monoazide (PMA) covalently bind with DNA molecules upon photolysis using visible light, and make it unavailable for DNA polymerase enzyme during polymerase chain reaction (PCR). Environmental DNA samples will be treated with suboptimum PMA concentration, enough to intercalate with 90 99% of the total DNA. The probability of PMA binding with DNA from abundant bacterial species will be much higher than binding with DNA from less abundant species. This will increase the relative DNA concentration of previously "shadowed" less abundant species available for PCR amplification. These PCR products obtained with and without PMA treatment will then be subjected to downstream diversity analyses such as sequencing and DNA microarray. It is expected that PMA-coupled PCR will amplify the "minority population" and help in understanding microbial diversity spectrum of an environmental sample at a much deeper level. This new protocol aims to overcome the major potential biases faced when analyzing microbial 16S rRNA gene diversity. This study will lead to a technological advancement and a commercial product that will aid microbial ecologists in understanding microbial diversity from various environmental niches. Implementation of this technique may lead to discoveries of novel microbes and their functions in sustenance of the ecosystem.

Vaishampayan, Parag A.↗