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NASA NTRS2008

Lab-on-a-Chip: From Astrobiology to the International Space Station

The continual and long-term habitation of enclosed environments, such as Antarctic stations, nuclear submarines and space stations, raises unique engineering, medical and operational challenges. There is no easy way out and no easy way to get supplies in. This situation elevates the importance of monitoring technology that can rapidly detect events within the habitat that affect crew safety such as fire, release of toxic chemicals and hazardous microorganisms. Traditional methods to monitor microorganisms on the International Space Station (ISS) have consisted of culturing samples for 3-5 days and eventual sample return to Earth. To augment these culture methods with new, rapid molecular techniques, we developed the Lab-on-a-Chip Application Development - Portable Test System (LOCAD-PTS). The system consists of a hand-held spectrophotometer, a series of interchangeable cartridges and a surface sampling/dilution kit that enables crew to collect samples and detect a range of biological molecules, all within 15 minutes. LOCAD-PTS was launched to the ISS aboard Space Shuttle Discovery in December 2006, where it was operated for the first time during March-May 2007. The surfaces of five separate sites in the US Lab and Node 1 of ISS were analyzed for endotoxin, using cartridges that employ the Limulus Amebocyte Lysate (LAL) assay; results of these tests will be presented. LOCAD-PTS will remain permanently onboard ISS with new cartridges scheduled for launch in February and October of 2008 for the detection of fungi (Beta-glucan) and Gram-positive bacteria (lipoteichoic acid), respectively.

Maule, Jake↗

NASA NTRS2006

Inter- and Intra-Chromosomal Aberrations in Human Cells Exposed in vitro to High and Low LET Radiations

Energetic heavy ions pose a health risk to astronauts in extended ISS and future Mars missions. High-LET heavy ions are particularly effective in causing various biological effects including cell inactivation, genetic mutations and cancer induction. Most of these biological endpoints are closely related to chromosomal damage, which can be utilized as a biomarker for radiation insults. Previously, we had studied chromosome aberrations in human lymphocytes and fibroblasts induced by both low- and high-LET radiation using FISH and multicolor fluorescence in situ hybridization (mFISH) techniques. In this study, we exposed human epithelial cells in vitro to gamma rays and energetic particles of varying types and energies and dose rates, and analyzed chromosomal damages using the multicolor banding in situ hybridization (mBAND) procedure. Confluent human epithelial cells (CH184B5F5/M10) were exposed to energetic heavy ions at NASA Space Radiation Laboratory (NSRL) at the Brookhaven National Laboratory, high energy neutron at the Los Alamos Nuclear Science Center (LANSCE) or Cs-137-gamma radiation source at the University of Texas, MD Anderson Cancer Center. After colcemid and Calyculin A treatment, cells were fixed and painted with XCyte3 mBAND kit (MetaSystems) and chromosome aberrations were analyzed with mBAND analysis system (MetaSystems). With this technique, individually painted chromosomal bands on one chromosome allowed the identification of interchromosomal aberrations (translocation to unpainted chromosomes) and intrachromosomal aberrations (inversions and deletions within a single painted chromosome). The results of the mBAND study showed a higher ratio of inversion involved with interchromosomal exchange in heavy ions compared to -ray irradiation. Analysis of chromosome aberrations using mBAND has the potential to provide useful information on human cell response to space-like radiation.

Hada, M.↗

NASA NTRS2012

Extreme Ionizing-Radiation-Resistant Bacterium

There is a growing concern that desiccation and extreme radiation-resistant, non-spore-forming microorganisms associated with spacecraft surfaces can withstand space environmental conditions and subsequent proliferation on another solar body. Such forward contamination would jeopardize future life detection or sample return technologies. The prime focus of NASA s planetary protection efforts is the development of strategies for inactivating resistance-bearing microorganisms. Eradification techniques can be designed to target resistance-conferring microbial populations by first identifying and understanding their physiologic and biochemical capabilities that confers its elevated tolerance (as is being studied in Deinococcus phoenicis, as a result of this description). Furthermore, hospitals, food, and government agencies frequently use biological indicators to ensure the efficacy of a wide range of radiation- based sterilization processes. Due to their resistance to a variety of perturbations, the non-spore forming D. phoenicis may be a more appropriate biological indicator than those currently in use. The high flux of cosmic rays during space travel and onto the unshielded surface of Mars poses a significant hazard to the survival of microbial life. Thus, radiation-resistant microorganisms are of particular concern that can survive extreme radiation, desiccation, and low temperatures experienced during space travel. Spore-forming bacteria, a common inhabitant of spacecraft assembly facilities, are known to tolerate these extreme conditions. Since the Viking era, spores have been utilized to assess the degree and level of microbiological contamination on spacecraft and their associated spacecraft assembly facilities. Members of the non-spore-forming bacterial community such as Deinococcus radiodurans can survive acute exposures to ionizing radiation (5 kGy), ultraviolet light (1 kJ/sq m), and desiccation (years). These resistive phenotypes of Deinococcus enhance the potential for transfer, and subsequent proliferation, on another solar body such as Mars and Europa. These organisms are more likely to escape planetary protection assays, which only take into account presence of spores. Hence, presences of extreme radiation-resistant Deinococcus in the cleanroom facility where spacecraft are assembled pose a serious risk for integrity of life-detection missions. The microorganism described herein was isolated from the surfaces of the cleanroom facility in which the Phoenix Lander was assembled. The isolated bacterial strain was subjected to a comprehensive polyphasic analysis to characterize its taxonomic position. This bacterium exhibits very low 16SrRNA similarity with any other environmental isolate reported to date. Both phenotypic and phylogenetic analyses clearly indicate that this isolate belongs to the genus Deinococcus and represents a novel species. The name Deinococcus phoenicis was proposed after the Phoenix spacecraft, which was undergoing assembly, testing, and launch operations in the spacecraft assembly facility at the time of isolation. D. phoenicis cells exhibited higher resistance to ionizing radiation (cobalt-60; 14 kGy) than the cells of the D. radiodurans (5 kGy). Thus, it is in the best interest of NASA to thoroughly characterize this organism, which will further assess in determining the potential for forward contamination. Upon the completion of genetic and physiological characteristics of D. phoenicis, it will be added to a planetary protection database to be able to further model and predict the probability of forward contamination.

Vaishampayan, Parag A.↗

NASA NTRS2013

Extreme Ionizing-Radiation-Resistant Bacterium

There is a growing concern that desiccation and extreme radiation-resistant, non-spore-forming microorganisms associated with spacecraft surfaces can withstand space environmental conditions and subsequent proliferation on another solar body. Such forward contamination would jeopardize future life detection or sample return technologies. The prime focus of NASA s planetary protection efforts is the development of strategies for inactivating resistance-bearing micro-organisms. Eradi cation techniques can be designed to target resistance-conferring microbial populations by first identifying and understanding their physiologic and biochemical capabilities that confers its elevated tolerance (as is being studied in Deinococcus phoenicis, as a result of this description). Furthermore, hospitals, food, and government agencies frequently use biological indicators to ensure the efficacy of a wide range of radiation-based sterilization processes. Due to their resistance to a variety of perturbations, the nonspore forming D. phoenicis may be a more appropriate biological indicator than those currently in use. The high flux of cosmic rays during space travel and onto the unshielded surface of Mars poses a significant hazard to the survival of microbial life. Thus, radiation-resistant microorganisms are of particular concern that can survive extreme radiation, desiccation, and low temperatures experienced during space travel. Spore-forming bacteria, a common inhabitant of spacecraft assembly facilities, are known to tolerate these extreme conditions. Since the Viking era, spores have been utilized to assess the degree and level of microbiological contamination on spacecraft and their associated spacecraft assembly facilities. Members of the non-sporeforming bacterial community such as Deinococcus radiodurans can survive acute exposures to ionizing radiation (5 kGy), ultraviolet light (1 kJ/m2), and desiccation (years). These resistive phenotypes of Deinococcus enhance the potential for transfer, and subsequent proliferation, on another solar body such as Mars and Europa. These organisms are more likely to escape planetary protection assays, which only take into account presence of spores. Hence, presences of extreme radiation-resistant Deinococcus in the cleanroom facility where spacecraft are assembled pose a serious risk for integrity of life-detection missions. The microorganism described herein was isolated from the surfaces of the cleanroom facility in which the Phoenix Lander was assembled. The isolated bacterial strain was subjected to a comprehensive polyphasic analysis to characterize its taxonomic position. This bacterium exhibits very low 16SrRNA similarity with any other environmental isolate reported to date. Both phenotypic and phylogenetic analyses clearly indicate that this isolate belongs to the genus Deinococcus and represents a novel species. The name Deinococcus phoenicis was proposed after the Phoenix spacecraft, which was undergoing assembly, testing, and launch operations in the spacecraft assembly facility at the time of isolation. D. phoenicis cells exhibited higher resistance to ionizing radiation (cobalt-60; 14 kGy) than the cells of the D. radiodurans (5 kGy). Thus, it is in the best interest of NASA to thoroughly characterize this organism, which will further assess in determining the potential for forward contamination. Upon the completion of genetic and physiological characteristics of D. phoenicis, it will be added to a planetary protection database to be able to further model and predict the probability of forward contamination.

Vaishampayan, Parag A.↗

NASA NTRS1976

Conceptual design of a biological specimen holding facility

An all-important first step in the development of the Spacelab Life Science Laboratory is the design of the Biological Specimen Holding Facility (BSHF) which will provide accommodation for living specimens for life science research in orbit. As a useful tool in the understanding of physiological and biomedical changes produced in the weightless environment, the BSHF will enable biomedical researchers to conduct in-orbit investigations utilizing techniques that may be impossible to perform on human subjects. The results of a comprehensive study for defining the BSHF, description of its experiment support capabilities, and the planning required for its development are presented. Conceptual designs of the facility, its subsystems and interfaces with the Orbiter and Spacelab are included. Environmental control, life support and data management systems are provided. Interface and support equipment required for specimen transfer, surgical research, and food, water and waste storage is defined. New and optimized concepts are presented for waste collection, feces and urine separation and sampling, environmental control, feeding and watering, lighting, data management and other support subsystems.

Jackson, J. K.↗

NASA NTRS2001

An Integrated Use of Experimental, Modeling and Remote Sensing Techniques to Investigate Carbon and Phosphorus Dynamics in the Humid Tropics

Moist tropical forests comprise one of the world's largest and most diverse biomes, and exchange more carbon, water, and energy with the atmosphere than any other ecosystem. In recent decades, tropical forests have also become one of the globe's most threatened biomes, subjected to exceptionally high rates of deforestation and land degradation. Thus, the importance of and threats to tropical forests are undeniable, yet our understanding of basic ecosystem processes in both intact and disturbed portions of the moist tropics remains poorer than for almost any other major biome. Our approach in this project was to take a multi-scale, multi-tool approach to address two different problems. First, we wanted to test if land-use driven changes in the cycles of probable limiting nutrients in forest systems were a key driver in the frequently observed pattern of declining pasture productivity and carbon stocks. Given the enormous complexity of land use change in the tropics, in which one finds a myriad of different land use types and intensities overlain on varying climates and soil types, we also wanted to see if new remote sensing techniques would allow some novel links between parameters which could be sensed remotely, and key biogeochemical variables which cannot. Second, we addressed to general questions about the role of tropical forests in the global carbon cycle. First, we used a new approach for quantifying and minimizing non-biological artifacts in the NOAA/NASA AVHRR Pathfinder time series of surface reflectance data so that we could address potential links between Amazonian forest dynamics and ENSO cycles. Second, we showed that the disequilibrium in C-13 exchanged between land and atmosphere following tropical deforestation probably has a significant impact on the use of 13-CO2 data to predict regional fluxes in the global carbon cycle.

Townsend, Alan R.↗

NASA NTRS2003

Physical association and functional interaction between beta1 integrin and CD98 on human T lymphocytes

CD98 is a cell surface protein previously characterized as a cell activation marker, an amino acid transporter, and has recently been implicated in integrin-related functions. Integrins are cell surface proteins, important for homotypic cell aggregation, cell adhesion, and coactivation of T lymphocytes. We have previously shown that the anti-CD98 mAb 80A10, when coimmobilized with anti-CD3 mAb OKT3, is able to mediate human T cell coactivation that is inhibited by anti-beta1 integrin specific mAb 18D3. These results indicated a functional association of CD98 and beta1 integrin signaling but left open the question of a physical association. We now show the induction of homotypic aggregation through CD98 among human T cells and this aggregation was inhibited by anti-beta1 integrin mAb. Therefore, CD98-dependent lymphocyte proliferation and adhesion may involve integrins. Competitive binding assays and fluorescence colocalization analysis suggested that CD98 and beta1 integrin were physically associated. Differential extraction techniques and immunoprecipitations provided the first evidence that the alpha4beta1 integrin and CD98 are specifically associated on human T lymphocytes.

NASA Discipline Cell Biology↗

NASA NTRS2006

Operation of a Cartesian Robotic System in a Compact Microscope with Intelligent Controls

A Microscope Imaging System (CMIS) with intelligent controls is disclosed that provides techniques for scanning, identifying, detecting and tracking microscopic changes in selected characteristics or features of various surfaces including, but not limited to, cells, spheres, and manufactured products subject to difficult-to-see imperfections. The practice of the present invention provides applications that include colloidal hard spheres experiments, biological cell detection for patch clamping, cell movement and tracking, as well as defect identification in products, such as semiconductor devices, where surface damage can be significant, but difficult to detect. The CMIS system is a machine vision system, which combines intelligent image processing with remote control capabilities and provides the ability to autofocus on a microscope sample, automatically scan an image, and perform machine vision analysis on multiple samples simultaneously.

McDowell, Mark↗

NASA NTRS2007

Tracking of cells with a compact microscope imaging system with intelligent controls

A Microscope Imaging System (CMIS) with intelligent controls is disclosed that provides techniques for scanning, identifying, detecting and tracking microscopic changes in selected characteristics or features of various surfaces including, but not limited to, cells, spheres, and manufactured products subject to difficult-to-see imperfections. The practice of the present invention provides applications that include colloidal hard spheres experiments, biological cell detection for patch clamping, cell movement and tracking, as well as defect identification in products, such as semiconductor devices, where surface damage can be significant, but difficult to detect. The CMIS system is a machine vision system, which combines intelligent image processing with remote control capabilities and provides the ability to auto-focus on a microscope sample, automatically scan an image, and perform machine vision analysis on multiple samples simultaneously.

McDowell, Mark↗

NASA NTRS2007

Tracking of Cells with a Compact Microscope Imaging System with Intelligent Controls

McDowell, Mark↗

NASA NTRS1997

Cytogenetic effects of space radiation in lymphocytes of MIR-18 crews

For assessing health risk, the measurement of physical dose received during a space mission, as well as the LETs, energies and charges of particles is important. It is also important to obtain quantitative information regarding the effectiveness of space radiation in causing damage to critical biological targets, e.g., chromosomes, since at present the estimated uncertainty of biological effects of space radiation is more than a factor of two. Such large uncertainty makes accurate health risk assessment very difficult. For this very reason, a study on cytogenetic effects of space radiation in human lymphocytes was proposed and done for MIR-18 mission. This study used FISH technique to score chromosomal translocations and C-banding method to determine dicentrics. Growth kinetics of cells and SCE were examined to ensure that chromosomal aberrations were scored in first mitosis and were induced not by chemical mutagens. Our results showed that chromosomal aberration frequency of post-flight samples was significantly higher than that of pre-flight ones and that SCE frequency was similar between pre- and post-flight samples. Based on a dose-response curve of preflight samples exposed to gamma rays, the absorbed dose received by crews during the mission was estimated to be about 14.5 cSv. Because the absorbed dose measured by physical dosimeters is 4.16 cGy for the entire mission, the RBE is about 3.5.

Mir Project↗

NASA NTRS2006

Identification Of Cells With A Compact Microscope Imaging System With Intelligent Controls

A Microscope Imaging System (CMIS) with intelligent controls is disclosed that provides techniques for scanning, identifying, detecting and tracking mic?oscopic changes in selected characteristics or features of various surfaces including, but not limited to, cells, spheres, and manufactured products subject to difficult-to-see imperfections. The practice of the present invention provides applications that include colloidal hard spheres experiments, biological cell detection for patch clamping, cell movement and tracking, as well as defect identification in products, such as semiconductor devices, where surface damage can be significant, but difficult to detect. The CMIS system is a machine vision system, which combines intelligent image processing with remote control capabilities and provides the ability to autofocus on a microscope sample, automatically scan an image, and perform machine vision analysis on multiple samples simultaneously.

McDowell, Mark↗

NASA NTRS1992

Episodes of terrestrial geologic activity during the past 260 million years - A quantitative approach

Records of major geologic events of the past about 260 Myr including: biologic extinction events, ocean-anoxic and black-shale events, major changes in sea level, major evaporite (salt) deposits, continental flood-basalt eruptions, first-order discontinuities in sea-floor spreading, and major mountain building events, have been aggregated and analyzed with moving-window and spectral techniques that facilitate recognition of clustering and possible cyclicity. Significant clustering of events suggests a model in which changes in rates and directions of sea-floor spreading ('ridge jumps') are associated with episodic rifting, volcanism, mountain building, global sea level and changes in the composition of the earth's atmosphere via the carbon cycle. The geologic data formally show a statistically significant underlying periodicity of 26.6 Myr for the Mesozoic and Cenozoic. Phase information suggests that the most recent maximum of the cycle occurred with the last 9 Myr, and may be close to the present time.

Rampino, Michael R.↗

NASA NTRS2007

Haughton-Mars Project/NASA 2006 Lunar Medical Contingency Simulation: Equipment and Methods for Medical Evacuation of an Injured Crewmember

Introduction: Achieving NASA's Space Exploration Vision scientific objectives will require human access into cratered and uneven terrain for the purpose of sample acquisition to assess geological, and perhaps even biological features and experiments. Operational risk management is critical to safely conduct the anticipated tasks. This strategy, along with associated contingency plans, will be a driver of EVA system requirements. Therefore, a medical contingency EVA scenario was performed with the Haughton-Mars Project/NASA to develop belay and medical evacuation techniques for exploration and rescue respectively. Methods: A rescue system to allow two rescuer astronauts to evacuate one in incapacitated astronaut was evaluated. The systems main components were a hard-bottomed rescue litter, hand-operated winch, rope, ground picket anchors, and a rover-winch attachment adapter. Evaluation was performed on 15-25deg slopes of dirt with embedded rock. The winch was anchored either by adapter to the rover or by pickets hammered into the ground. The litter was pulled over the surface by rope attached to the winch. Results: The rescue system was utilized effectively to extract the injured astronaut up a slope and to a waiting rover for transport to a simulated habitat for advanced medical care, although several challenges to implementation were identified and overcome. Rotational stabilization of the winch was found to be important to get maximize mechanical advantage from the extraction system. Discussion: Further research and testing needs to be performed to be able to fully consider synergies with the other Exploration surface systems, in conducting contingency operations. Structural attachment points on the surface EVA suits may be critical to assist in incapacitated evacuation. Such attach points could be helpful in microgravity incapacitated crewmember transport as well. Wheeled utility carts or wheels that may be attachable to a litter may also aid in extraction and transport. Utilizing parts of the rover (e.g. seats) to deploy as a litter may be considered. Testing in simulated 1/6-g to determine feasibility of winch operation and anchor establishment will further reduce implementation uncertainties.

Chappell, S. P.↗

NASA NTRS1988

Growth inhibition, turgor maintenance, and changes in yield threshold after cessation of solute import in pea epicotyls

The dependence of stem elongation on solute import was investigated in etiolated pea seedlings (Pisum sativum L. var Alaska) by excising the cotyledons. Stem elongation was inhibited by 60% within 5 hours of excision. Dry weight accumulation into the growing region stopped and osmotic pressure of the cell sap declined by 0.14 megapascal over 5 hours. Attempts to assay phloem transport via ethylenediaminetetraacetate-enhanced exudation from cut stems revealed no effect of cotyledon excision, indicating that the technique measured artifactual leakage from cells. Despite the drop in cell osmotic pressure, turgor pressure (measured directly via a pressure probe) did not decline. Turgor maintenance is postulated to occur via uptake of solutes from the free space, thereby maintaining the osmotic pressure difference across the cell membrane. Cell wall properties were measured by the pressure-block stress relaxation technique. Results indicate that growth inhibition after cotyledon excision was mediated primarily via an increase in the wall yield threshold.

NASA Discipline Number 40-30↗

NASA NTRS1973

An evaluation of space acquired data as a tool for management to wildlife habitat in Alaska

The Bureau of Sport Fisheries and Wildlife ERTS experiment in Alaska attempts to yield information useful for three primary functions in the State. They are: (1) to test the feasibility of using ERTS data, in conjunction with aircraft acquired multispectral photography, to develop effective stratified sampling techniques, (2) to provide near real time assessment and evaluation of the quantity, quality, and distribution of waterfowl breeding habitat through frequent ERTS measurements of hydrologic, phenological and vegetational parameters, and (3) to provide basic mapping of vegetation and terrain in certain remote areas of the State for which little or no biological data now exist.

Vantries, B. J.↗

NASA NTRS1991

BOREAS (Boreal Ecosystem-Atmosphere Study): Global change and biosphere-atmosphere interactions in the boreal forest

The Boreal Ecosystems Atmosphere Study (BOREAS) is a cooperative field and analysis project involving elements of land surface climatology, tropospheric chemistry, and terrestrial ecology. The goal of the study is to understand the interactions between the boreal forest biome and the atmosphere in order to clarify their roles in global change. The study will be centered on two 20 by 20 km sites within the North American boreal forest region, located near the northern and southern limits of the biome. Studies based at these sites will be used to explore the roles of various environmental factors in controlling the extent and character of the biome. The sites will be the subject of surface, airborne, and satellite based observations which aim to improve understanding of the biological and physical processes and states which govern the exchanges of energy, water, carbon, and trace gases between boreal forest ecosystems and the atmosphere. Particular reference will be made to those processes and states that may be sensitive to global change. The study also aims to develop the use of remote sensing techniques to transfer understanding of the above process from local scales out to regional scales. The BOREAS project is being planned for 1992-1996, with a major field effort in 1994.

Sellers, Piers J.↗

NASA NTRS2001

Cell-cycle research with synchronous cultures: an evaluation

The baby-machine system, which produces new-born Escherichia coli cells from cultures immobilized on a membrane, was developed many years ago in an attempt to attain optimal synchrony with minimal disturbance of steady-state growth. In the present article, we put forward a model to describe the behaviour of cells produced by this method, and provide quantitative evaluation of the parameters involved, at each of four different growth rates. Considering the high level of selection achievable with this technique and the natural dispersion in interdivision times, we believe that the output of the baby machine is probably close to optimal in terms of both quality and persistence of synchrony. We show that considerable information on events in the cell cycle can be obtained from populations with age distributions very much broader than those achieved with the baby machine and differing only modestly from steady state. The data presented here, together with the long and fruitful history of findings employing the baby-machine technique, suggest that minimisation of stress on cells is the single most important factor for successful cell-cycle analysis.

Non-NASA Center↗