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NASA NTRS1999

The use of a spaceflight-compatible device to perform WBC surface marker staining and whole-blood mitogenic activation for cytokine detection by flow cytometry

Significant changes have recently been described regarding circulating peripheral immune cells immediately following spaceflight. Existing methods for immunophenotype staining of peripheral blood in terrestrial labs do not meet the constraints for flight on the Space Shuttle. We have recently described the development and use of the Whole Blood Staining Device (WBSD), a simple device for staining flow cytometry specimens during spaceflight. When preparing samples with the WBSD, all liquids are safely contained as the cells are moved through staining, lysis and fixation steps. Here we briefly review the use of the WBSD, and then describe another versatile adaptation, a modification to perform intracellular staining of cytokines for detection by flow cytometry. Alterations in cytokine production have been reported both in ground-based simulated microgravity culture and in astronaut samples returning from spaceflight. Data regarding microgravity effects on cytokine production for specific subpopulations of cells is lacking. Flow cytometric cytokine analysis offers the unique ability to perform simultaneous surface marker analysis and positively identity cytokine producing subsets of cells. The utilization of the WBSD provides the ability to perform rapid and routine mitogenic activation during spaceflight coupled with the ability to perform simultaneous surface marker analysis. The only external requirements for this procedure are an in-flight 37-degree incubator and the capacity for 4-degree storage.

NASA Discipline Regulatory Physiology↗

NASA NTRS2006

Improving the Near-Earth Micrometeoroid and Orbital Debris Environment Definition with LADC

The Large Area Debris Collector (LADC) is a 10 m(sup 2) aerogel and acoustic sensor system designed to characterize and collect submillimeter micrometeoroids and orbital debris on the International Space Station (ISS). The project is led by the U.S. Naval Research Laboratory (NRL) with major collaboration by the NASA Orbital Debris Program Office at Johnson Space Center. The U.S. Department of Defense Space Test Program (STP) is responsible for the integration, deployment, and retrieval of the system. The deployment is scheduled for August 2007 with an orbital collection period of one to two years. The combined area time product of LADC will provide a much needed orbital debris population update in the size regime that is important to the safety community - 100 mm and larger. Another key element for LADC is the source identification of the collected samples. Impact features such as track length and track volume can be used to estimate the impact speed and direction of any selected residual embedded in aerogel. Acoustic sensors can provide impact timing and impact location information. The combined dynamical signatures make it possible to reconstruct the orbits of some of the collected samples and lead to their source identification. Compositional analysis on the residuals can also separate debris from meteoroids and provide additional population breakdown for orbital debris (e.g., Al, paint, steel, Al2O3). To maximize the science return and minimize potential contamination from other ISS modules, a careful selection of the location and orientation of LADC on the ISS is needed. Key issues and engineering constraints encountered during mission preparation, and the expected science return based on the mission configuration, are summarized in this paper.

Liou, J.-C.↗

NASA NTRS2020

Recommended Laboratory and Field Studies ahead of Future Mercury Exploration

Data from Mariner 10, MESSENGER, and ground-based telescopic observations have facilitated great advancements towards understanding the geochemistry, geology, internal structure, and space environment (exosphere/magnetosphere) of Mercury. Future exploration efforts by BepiColombo, landed science, and sample return will revolutionize our understanding of this planet. However, to enhance the scientific return of past, present, and future data collection at Mercury, it is imperative that laboratory and field-based studies be conducted over the next decade. This white paper gives examples of laboratory and field studies, including instrument testing, which should take place over the next decade to facilitate the interpretation of data and preparation of future missions to the inner-most planet. By no means are the topics in this paper exhaustive, but rather are meant to demonstrate the immense amount of work that needs to be completed ahead of, or in conjunction with, current and future exploration efforts of Mercury. This work would amplify the benefits from additional data obtained from the planet’s surface to the fullest extent possible, and to aid in decision making of future instrument payloads. We have chosen to focus our inputs here on geochemistry, geology, and space environment related efforts, but we emphasize that extensive research in the areas of geophysics and other geosciences-related disciplines will benefit from additional studies.

Kathleen E. Vander Kaaden↗

NASA NTRS2010

Curating NASA's Extraterrestrial Samples - Past, Present, and Future

Curation of extraterrestrial samples is the critical interface between sample return missions and the international research community. The Astromaterials Acquisition and Curation Office at the NASA Johnson Space Center (JSC) is responsible for curating NASA's extraterrestrial samples. Under the governing document, NASA Policy Directive (NPD) 7100.10E "Curation of Extraterrestrial Materials," JSC is charged with ". . . curation of all extraterrestrial material under NASA control, including future NASA missions." The Directive goes on to define Curation as including documentation, preservation, preparation, and distribution of samples for research, education, and public outreach.

Allen, Carlton↗

NASA NTRS2011

Curating NASA's Extraterrestrial Samples - Past, Present, and Future

Curation of extraterrestrial samples is the critical interface between sample return missions and the international research community. The Astromaterials Acquisition and Curation Office at the NASA Johnson Space Center (JSC) is responsible for curating NASA s extraterrestrial samples. Under the governing document, NASA Policy Directive (NPD) 7100.10E "Curation of Extraterrestrial Materials", JSC is charged with ". . . curation of all extraterrestrial material under NASA control, including future NASA missions." The Directive goes on to define Curation as including "documentation, preservation, preparation, and distribution of samples for research, education, and public outreach."

Allen, Carlton↗

NASA NTRS2023

An Engineering Guide to Lunar Geotechnical Properties

The renewed interest in returning human and robotic explorers to the lunar surface has identified a need for a renewed understanding of lunar geotechnical properties related to landing, exploration, excavation, and construction activities on the lunar surface. This paper summarizes measurements conducted during US and Russian/Soviet landed missions as well as experiments performed on returned samples to establish fundamental geotechnical properties such as particle size distribution, particle shape, bulk density, shear strength, cohesion and bearing strength. While many of these properties are well known, how they vary with increased lunar soil depth is less understood, and those properties that vary significantly as a function of depth are explored in additional detail. Selected examples discuss mechanical excavation forces, rocket exhaust erosion forces, and the preparation of launch/landing pad surfaces, with the goal of a better understanding of lunar soil geotechnical properties that apply to large-scale exploration of the lunar surface and dictate the design of future exploration systems.

Lunar Soil↗

NASA NTRS2008

Pattern-Recognition System for Approaching a Known Target

A closed-loop pattern-recognition system is designed to provide guidance for maneuvering a small exploratory robotic vehicle (rover) on Mars to return to a landed spacecraft to deliver soil and rock samples that the spacecraft would subsequently bring back to Earth. The system could be adapted to terrestrial use in guiding mobile robots to approach known structures that humans could not approach safely, for such purposes as reconnaissance in military or law-enforcement applications, terrestrial scientific exploration, and removal of explosive or other hazardous items. The system has been demonstrated in experiments in which the Field Integrated Design and Operations (FIDO) rover (a prototype Mars rover equipped with a video camera for guidance) is made to return to a mockup of Mars-lander spacecraft. The FIDO rover camera autonomously acquires an image of the lander from a distance of 125 m in an outdoor environment. Then under guidance by an algorithm that performs fusion of multiple line and texture features in digitized images acquired by the camera, the rover traverses the intervening terrain, using features derived from images of the lander truss structure. Then by use of precise pattern matching for determining the position and orientation of the rover relative to the lander, the rover aligns itself with the bottom of ramps extending from the lander, in preparation for climbing the ramps to deliver samples to the lander. The most innovative aspect of the system is a set of pattern-recognition algorithms that govern a three-phase visual-guidance sequence for approaching the lander. During the first phase, a multifeature fusion algorithm integrates the outputs of a horizontal-line-detection algorithm and a wavelet-transform-based visual-area-of-interest algorithm for detecting the lander from a significant distance. The horizontal-line-detection algorithm is used to determine candidate lander locations based on detection of a horizontal deck that is part of the lander.

Huntsberger, Terrance↗

NASA NTRS2005

Development and Test Plans for the MSR EEV

The goal of the proposed Mars Sample Return mission is to bring samples from the surface of Mars back to Earth for thorough examination and analysis. The Earth Entry Vehicle is the passive entry body designed to protect the sample container from entry heating and deceleration loads during descent through the Earth s atmosphere to a recoverable location on the surface. This paper summarizes the entry vehicle design and outlines the subsystem development and testing currently planned in preparation for an entry vehicle flight test in 2010 and mission launch in 2013. Planned efforts are discussed for the areas of the thermal protection system, vehicle trajectory, aerodynamics and aerothermodynamics, impact energy absorption, structure and mechanisms, and the entry vehicle flight test.

Dillman, Robert↗

NASA NTRS2004

Forward Contamination of the Moon and Mars: Implications for Future Life Detection Missions

NASA and ESA have outlined new visions for solar system exploration that will include a series of lunar robotic missions to prepare for, and support a human return to the Moon, and future human exploration of Mars and other destinations. One of the guiding principles for exploration is to pursue compelling scientific questions about the origin and evolution of life. The search for life on objects such as Mars will require that all spacecraft and instrumentation be sufficiently cleaned and sterilized prior to launch to ensure that the scientific integrity of extraterrestrial samples is not jeopardized by terrestrial organic contamination. Under COSPAR's current planetary protection policy for the Moon, no sterilization procedures are required for outbound lunar spacecraft. Nonetheless, future in situ investigations of a variety of locations on the Moon by highly sensitive instruments designed to search for biologically derived organic compounds would help assess the contamination of the Moon by lunar spacecraft. These studies could also provide valuable "ground truth" data for Mars sample return missions and help define planetary protection requirements for future Mars bound spacecraft carrying life detection experiments. In addition, studies of the impact of terrestrial contamination of the lunar surface by the Apollo astronauts could provide valuable data to help refine future Mars surface exploration plans for a human mission to Mars.

Glavin, Daniel P.↗

NASA NTRS2010

In Situ Biological Contamination Studies of the Moon: Implications for Future Planetary Protection and Life Detection Missions

NASA and ESA have outlined visions for solar system exploration that will include a series of lunar robotic precursor missions to prepare for, and support a human return to the Moon, and future human exploration of Mars and other destinations. One of the guiding principles for exploration is to pursue compelling scientific questions about the origin and evolution of life. The search for life on objects such as Mars will require that all spacecraft and instrumentation be sufficiently cleaned and sterilized prior to launch to ensure that the scientific integrity of extraterrestrial samples is not jeopardized by terrestrial organic contamination. Under the Committee on Space Research's (COSPAR's) current planetary protection policy for the Moon, no sterilization procedures are required for outbound lunar spacecraft, nor is there yet a planetary protection category for human missions. Future in situ investigations of a variety of locations on the Moon by highly sensitive instruments designed to search for biologically derived organic compounds would help assess the contamination of the Moon by lunar spacecraft. These studies could also provide valuable "ground truth" data for Mars sample return missions and help define planetary protection requirements for future Mars bound spacecraft carrying life detection experiments. In addition, studies of the impact of terrestrial contamination of the lunar surface by the Apollo astronauts could provide valuable data to help refine future Mars surface exploration plans for a human mission to Mars.

Glavin, Daniel P.↗

NASA NTRS2020

Mars 2020: Mission, Science Objectives and Build

If all goes according to plan, in February 2021, NASA will land the Mars 2020 Rover on the surface of Mars. Mars 2020 is the latest in a series of unmanned Martian robotic rover missions that are part of NASA’s Mars Exploration Program, a long-term effort of robotic exploration of the planet. The mission seeks to address high-priority goals for Mars exploration, including answering questions about the potential for past life on Mars. Mars 2020 will look for evidence of habitable conditions on Mars in the ancient past, as well as look for signs of past microbial life itself. The mission also seeks to understanding the geological history and evolution of the planet, and to prepare for future robotic and human exploration. The Mars 2020 spacecraft and rover borrow heavily from the Mars Science Laboratory (MSL) mission and Curiosity rover which landed on Mars in 2012. This reliance on proven technology helps reduce mission risk and cost. Mars 2020 does contain new technology, including a drill for coring samples from Martian rock and soil and a Sample Caching System for gathering, storing and preserving samples for possible future return to Earth. In this paper, we will review the primary goals of the Mars 2020 Mission and look at the reasons for choosing Jezero Crater as the landing site. We will discuss the design and build of the Mars 2020 Spacecraft system and its similarities and differences with Mars Science Laboratory and the Curiosity Rover. We will also review the Mars 2020 Scientific Instrument Suite and their goals. Finally, we will review the Return Sample Contamination Control requirements and the design choices that were made to facilitate meeting these requirements.

Soares, Carlos E.↗

NASA NTRS2016

Successful Validation of Sample Processing and Quantitative Real-Time PCR Capabilities on the International Space Station

The WetLab-2 system was developed by NASA Ames Research Center to offer new capabilities to researchers. The system can lyse cells and extract RNA (Ribonucleic Acid) on-orbit from different sample types ranging from microbial cultures to animal tissues. The purified RNA can then either be stabilized for return to Earth or can be used to conduct on-orbit quantitative Reverse Transcriptase PCR (Polymerase Chain Reaction) (qRT-PCR) analysis without the need for sample return. The qRT-PCR results can be downlinked to the ground a few hours after the completion of the run. The validation flight of the WetLab-2 system launched on SpaceX-8 on April 8, 2016. On orbit operations started on April 15th with system setup and was followed by three quantitative PCR runs using an E. coli genomic DNA template pre-loaded at three different concentrations. These runs were designed to discern if quantitative PCR functions correctly in microgravity and if the data is comparable to that from the ground control runs. The flight data showed no significant differences compared to the ground data though there was more variability in the values, this was likely due to the numerous small bubbles observed. The capability of the system to process samples and purify RNA was then validated using frozen samples prepared on the ground. The flight data for both E. coli and mouse liver clearly shows that RNA was successfully purified by our system. The E. coli qRT-PCR run showed successful singleplex, duplex and triplex capability. Data showed high variability in the resulting Cts (Cycle Thresholds [for the PCR]) likely due to bubble formation and insufficient mixing during the procedure run. The mouse liver qRT-PCR run had successful singleplex and duplex reactions and the variability was slightly better as the mixing operation was improved. The ability to purify and stabilize RNA and to conduct qRT-PCR on-orbit is an important step towards utilizing the ISS as a National Laboratory facility. The ability to get on-orbit data will provide investigators with the opportunity to adjust experimental parameters in real time without the need for sample return and re-flight. The WetLab-2 Project is supported by the Research Integration Office in the ISS Program.

gene expression↗

NASA NTRS2016

Successful Validation of Sample Processing and Quantitative Real-Time PCR Capabilities on the International Space Station

quantitative PCR↗

NASA NTRS2010

Permissible Exposure Level for Lunar Dusts: Gaps are Closing

Space faring nations plan to return human explorers to the moon within the next decade. Experience during the Apollo flights suggests that lunar dust will invariably get into the habitat where the finest portion (less than 5 micrometers) could be inhaled by the crew before it is cleared from the atmosphere. NASA is developing a database from which a 6-month, episodic exposure standard for lunar dust can be set. Three kinds of moon dust were prepared from a parent sample of Apollo 14 regolith #14003,96. Our goal was to prepare each type of dust sample with a mean diameter less than 2 m, which is suitable for instillation into the lungs of rats. The three samples were prepared as follows: separation from the parent sample using a fluidized bed, grinding using a jet mill grinder, or grinding with a ball-mill grinder. Grinding simulated restoration of surface activation of dust expected to occur at the surface of the moon on native lunar dust. We used two grinding methods because they seemed to produce different modes of activation. The effects of grinding were preserved by maintaining the dust in ultra-pure nitrogen until immediately before it was placed in suspension for administration to rats. The dust was suspended in physiological saline with 10% Survanta, a lung surfactant. Rats were given intratrachael instillations of the dust suspension at three doses. In addition to the three moon dusts (A, C and E), we instilled the same amount of a negative control (TiO2, B) and a highly-toxic, positive control (quartz, D). These additional mineral dusts were selected because they have well-established and very different permissible exposure levels (PELs). Our goal was to determine where lunar dusts fit between these extremes, and then estimate a PEL for each lunar dust. We evaluated many indices of toxicity to the lung. The figure shows the changes in lactate dehydrogenase (LDH), a marker of cell death, for the five dusts. Benchmark dose software (Version 2.1.2) from the Environmental Protection Agency was used to estimate the 10% effect levels (BMD(sub 10)) using five models. The best-fitting model was used to estimate the optimal BMD(sub 10) (table)

James, John T.↗

NASA NTRS2010

In Situ Biological Contamination Studies of the Moon: Implications for Planetary Protection and Life Detection Missions

NASA and ESA have outlined visions for solar system exploration that will include a series of lunar robotic precursor missions to prepare for, and support a human return to the Moan, and future human exploration of Mars and other destinations, including possibly asteroids. One of the guiding principles for exploration is to pursue compelling scientific questions about the origin and evolution of life. The search for life on objects such as Mars will require careful operations, and that all systems be sufficiently cleaned and sterilized prior to launch to ensure that the scientific integrity of extraterrestrial samples is not jeopardized by terrestrial organic contamination. Under the Committee on Space Research's (COSPAR's) current planetary protection policy for the Moon, no sterilization procedures are required for outbound lunar spacecraft, nor is there a different planetary protection category for human missions, although preliminary C SPAR policy guidelines for human missions to Mars have been developed. Future in situ investigations of a variety of locations on the Moon by highly sensitive instruments designed to search for biologically derived organic compounds would help assess the contamination of the Moon by lunar spacecraft. These studies could also provide valuable "ground truth" data for Mars sample return missions and help define planetary protection requirements for future Mars bound spacecraft carrying life detection experiments. In addition, studies of the impact of terrestrial contamination of the lunar surface by the Apollo astronauts could provide valuable data to help refine future: Mars surface exploration plans for a human mission to Mars.

Glavin, Daniel P.↗

NASA NTRS2016

Mars 2020 Rover Mission Status in 2016

Mars 2020, NASA’s next Mars rover mission, continues NASA’s Mars Exploration Program with a focus on seeking signs of ancient life and helping to prepare for future human explorers. NASA will select a landing site likely to have once been habitable by microbial life and with the geologic record capable of preserving signs of its presence, if it ever existed. The rover will explore the landing site using a powerful set of instruments that support both in situ investigations and the selection of Mars samples to be collected, sealed, and left on the surface of Mars for possible future return.

Bernard, Douglas↗

NASA NTRSDate not supplied

Culture-Independent Microbial Air Profiling using a Spaceflight-Compatible Nanopore Sequencing Method

Microbial monitoring of spacecraft air is critical toward assessing the efficacy of microbial controls within the environmental control and life support systems to protect the crew and vehicle environment. Currently, onboard the International Space Station (ISS), the air is monitored every quarter using an impaction air sampler. With this method, microbial cells and spores are pulled onto plates containing a growth medium. Following onboard incubation, the crew reports approximate microbial levels to the ground, but sample return is required for identification. Upon return of the plates, the isolates present are identified for crew health risk assessments. As NASA moves beyond low-Earth orbit, sample return will be impractical, and a near real-time monitoring capability is essential. Significant strides have been made in recent years to utilize a molecular-based method for microbial profiling of ISS surfaces. The developed method is independent of microbial culture, thus removing the bias toward detecting only culturable organisms, eliminating the need for sample return, and reducing the risk to crew health from exposure to high microbial levels. The work described here details the evaluation of three different air sampling platforms whose product is amenable to downstream molecular processing. The three samplers were compared in terms of mass and power requirements, ease of use, and the resulting data. For the two highest-ranking samplers, a basic concept of operations was developed to transfer the sample into the already established preparation and sequencing process. Using these concepts of operations, an in-depth comparison of the molecular data generated was compared to the historical culture-based method. Data from both methods detailed similar microbial profiles, while the molecular method detailed microbial identifications that were lacking from the culture data. The developed method will enable the generation of near real-time microbial profiles of the spacecraft atmosphere.

Brandon Dunbar↗

NASA NTRS2008

Rapid Biochemical Analysis on the International Space Station (ISS): Preparing for Human Exploration of the Moon and Mars

The Lab-on-a-Chip Application Development - Portable Test System, known as LOCAD-PTS, was launched to the International Space Station (ISS) aboard Space Shuttle Discovery (STS-116) on December 9th,2006. Since that time, it has remained onboard ISS and has been operated by the crew on 10 separate occasions LOCAD-PTS is a handheld device for rapid biochemical analysis; it consists of a spectrophotometer, a series of interchangeable cartridges, a pipette and several clean/sterilized swabbing kits to obtain samples from ISS surfaces. Sampling, quantitative analysis and data retrieval is performed onboard, therefore reducing the need to return samples to Earth. Less than 20 minutes are required from sampling to data, significantly faster than existing culture-based methods on ISS, which require 3-5 days. Different cartridges are available for the detection of different target molecules (simply by changing the formulation within each cartridge), thereby maximizing the benefit and applications addressed by a single instrument. Initial tests on ISS have focused on the detection of the bact.erial macromolecule endotoxin, a component of bacterial cell walls. LOCAD-PTS detects endotoxin with a cartridge that contains a formulation known as Limulus Amebocyte Lysate (LAL) assay. LAL is derived from blood of the horseshoe crab, Limulus polyphemus, and detects enodotoxin with an enzyme cascade that triggers generation Of a yellow colored dye, p-nitroanaline. The more p-nitroanaline product, the more endotoxin is in the original sample. To enable quantitative analysis, the absorbance of this color is measured by LOCAD-PTS through a 395 nm filter and compared with an internal calibration curve, to provide a reading on the LED display that ranges from 0.05 Endotoxin Units (EU)/ml to 5 EU/ml. Several surface sites were analyzed within ISS between March 2007 and February 2008, including multiple locations in the US Laboratory Destiny, Node 1 Unity, AMock, and Service Module Zvezda. The goals of this initial study were to i) test the cleanliness of reagents/supplies on orbit, ii) test the crew's ability to collect and process a sample in microgravity without contamination, iii) demonstrate nominal function of the LOCAD-PTS, and iv) provide a general survey of endotoxin within the ISS. The surface sites varied greatly in terms of their frequency-of-use and material texture/composition; from relatively smooth aluminum, to fabric, to the room temperature vulcanizing (RTV) rubber of a Extravehicular Mobility Unit (EMU) spacesuit. Results showed that: i) the swabbing kits and reagents remained clean on orbit, ii) the crew could collect and process a sample without contamination, and iii) the LOCAD-PTS functioned nominally in > 99% of the 55 tests completed. We will present detailed results of the survey of endotoxin on ISS surfaces. These results and technology are important in the near-term - by providing an extra tool in the toolbox for ISS microbial monitoring. They are also important in the longer term as valuable preparation for human exploration of the Moon and Mars. One of the proposed science goals for the human exploration of Mars will be to detect and characterize any indigenous biological molecules that may exist on the Martian surface. To achieve that goal, the crew must have the technology available onboard to differentiate indigenous biology from any terrestrial biological material brought to Mars by the spacecraft and crew (termed 'forward contamination'). The LAL assay is already one of the official methods used by NASA's planetary protection program to certify cleanliness of interplanetary robotic spacecraft prior to launch; and therefore endotoxin is a good marker of forward contamination (as well as other microbial molecules detectable with LOCAD-PTS e.g. box-1, 3-glucan and lipoteichoic acid). Furthermore, the distribution and abundance of these molecules on the ISS provides a good indicator of what to expect on the Crew Exploratioehicle Orion, the lunar lander Antares, and future crewed spacecraft destined for Mars. In addition, technology such as LOCAD-PTS has been proposed to help evaluate forward contamination during lunar surface operations by the crew, as preparation for the human exploration of Mars.

Maule, J.↗