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At least 91 records · Page 5

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NASA NTRS1976

Development of a preliminary design of a method to measure the effectiveness of virus exclusion during water process reclamation at zero-G

Organon Diagnostics has developed, under NASA sponsorship, a monitoring system to test the capability of a water recovery system to reject the passage of viruses into the recovered water. In this system, a non-pathogenic marker virus, bacteriophage F2, is fed into the process stream before the recovery unit and the reclaimed water is assayed for its presence. An engineering preliminary design has been performed as a parallel effort to the laboratory development of the marker virus test system. Engineering schematics and drawings present a preliminary instrument design of a fully functional laboratory prototype capable of zero-G operation.

Fraser, A. S.↗

NASA NTRS1994

Monoclonal antibodies directed against surface molecules of multicell spheroids

The objective of this project is to generate a library of monoclonal antibodies (MAbs) directed against surface molecules of tumor and transformed cells grown as multicell spheroids (MCS). These MCS are highly organized, 3-dimensional multicellular structures which exhibit many characteristics of in vivo organized tissues not found in conventional monolayer or suspension culture. Therefore MCS make better in vitro model systems to study the interactions of mammalian cells, and provide a functional assay for surface adhesion molecules. This project also involves investigations of cell-cell interactions in a gravity-based environment. It will provide a base of scientific information necessary to expand the focus of the project in future years to microgravity and hypergravity-based environments. This project also has the potential to yield important materials (e.g., cellular products) which may prove useful in the diagnosis and/or treatment of certain human diseases. Moreover, this project supports the training of both undergraduate and graduate students; thus, it will assist in developing a pool of future scientists with research experience in an area (gravitational biology) of interest to NASA.

Martinez, Andrew O.↗

NASA NTRS2006

The Fourier-Kelvin Stellar Interferometer a Low Complexity, Low Cost Space Mission for High-Resolution Astronomy and Direct Exoplanet Detection

The Fourier-Kelvin Stellar Interferometer (FKSI) is a mission concept for a spacecraft-borne nulling interferometer for high-resolution astronomy and the direct detection of exoplanets and assay of their environments and atmospheres. FKSI is a high angular resolution system operating in the near to midinfrared spectral region and is a scientific and technological pathfinder to the Darwin and Terrestrial Planet Finder (TPF) missions. The instrument is configured with an optical system consisting, depending on configuration, of two 0.5 - 1.0 m telescopes on a 12.5 - 20 m boom feeding a symmetric, dual Mach- Zehnder beam combiner. We report on progress on our nulling testbed including the design of an optical pathlength null-tracking control system and development of a testing regime for hollow-core fiber waveguides proposed for use in wavefront cleanup. We also report results of integrated simulation studies of the planet detection performance of FKSI and results from an in-depth control system and residual optical pathlength jitter analysis.

Barry, R. K.↗

NASA NTRSDate not supplied

Application of Analytical Hierarchy Process for Narrowing Down Nep Candidate Reactor Designs

Nuclear electric propulsion (NEP)-powered vehicles have been contemplated for human Mars missions. The nuclear power system contemplates using a high temperature light-weight nuclear reactor for the production of electrical power in the range of 2-5 MWe with a 3-10 year service life. A myriad of technology options exist for achieving these mission objectives. The analytic hierarchy process (AHP), a multi-attribute decision method, is being used to narrow down the candidate designs. The AHP is a structured decision process that fuses model-supplied quantitative data with subjective assessments to facilitate decisions that involve multiple competing criteria. Proven end-to-end nuclear design and systems analysis tools will be used to provide quantitative performance data such as end-to-end system reliability, system robustness to recover from off-normal conditions, system specific weight (α in kg/kWe), and the ability to meet the service life-time and power level requirements. Three high assay low enriched uranium (HALEU) reactor concepts, namely, gas-, heat pipe- and pumped liquid-cooled nuclear cores coupled to a He/Xe gas or supercritical CO2- Brayton power conversion system are being modeled using the AHP to understand the trade-offs associated with these design combinations. In addition to the reactor and power conversion options, there are a myriad of additional components and subsystems – e.g., radiators, heat exchangers, and recuperators – that also figure into the evaluation process. The choice of the right overall system is a multidimensional problem that has to include not only quantitative data, but also the so called “external factors”, examples of which include the component and subsystem Technology Readiness Levels (TRLs), the associated Advancement Degrees of Difficulty (AD2), the cost and schedule required to achieve a technical maturity consistent with mission infusion, alignment with the priorities of NASA’s Space Nuclear Propulsion program, and alignment with other ongoing government and commercial investments in micro reactors. Focused expert elicitations form the basis for qualitative data set. As a final step, decision-makers individually express their opinions regarding the relative importance of the criteria and preferences among the alternatives through pairwise comparisons. The paper will describe the AHP approach, progress to-date applying it to the NEP human Mars mission problem, and preliminary results. Use of AHP provides sufficient flexibility for incorporating industry input at different stages as the technologies evolve through additional research and development. It is expected that the decision process will be ongoing and expanded to examine additional options and technology choices, culminating in a defensible set of candidate reactor concepts that will form the basis for developing a multi-year NEP technology maturation strategy. .

Dasari V Rao↗

NASA NTRS1993

Monoclonal antibodies directed against surface molecules of multicell spheroids

The objective of this project is to generate a library of monoclonal antibodies (MAbs) to surface molecules of mammalian tumor and transformed cells grown as multicell spheroids (MCS). These MCS are highly organized, three dimensional multicellular structures which exhibit many characteristics of in vivo organized tissues not found in conventional monolayer or suspension culture; therefore, MCS make better in vitro model systems to study the interactions of mammalian cells. Additionally, they provide a functional assay for surface adhesion molecules.

Martinez, Andrew O.↗

NASA NTRS1994

Monoclonal antibodies directed against surface molecules of multicell spheroids

The objective of this project is to generate a library of monoclonial antibodies (MAbs) directed against surface molecules of tumor and transformed cells grown as multicell spheroids (MCS). These MCS are highly organized, 3-dimensional multicellular structures which exhibit many characteristics of in vivo organized tissues which are not found in conventional monolayer or suspension culture. In brief, MCS combine the relevance or organized tissues with in vitro methodology making the MCS a good model system to study the interactions of mammalian cells, and thereby provide a functional assay for surface adhesion molecules. This project also involves investigations of cell-cell interactions in a gravity-based environment. It will provide an important base of scientific information for future comparative studies on the effects of hypergravity and simulated microgravity environments on cell-cell interactions. This project also has the potential to yield important materials (e.g. cellular products) which may be useful for the diagnosis and/or treatment of certain human diseases. Moreover, this project supports the training of one undergraduate and one graduate student; thus, it will also assist in developing a pool of future scientists with research experience in gravitational biology research.

Martinez, Andrew O.↗

NASA NTRS2006

Studying Functions of All Yeast Genes Simultaneously

A method of studying the functions of all the genes of a given species of microorganism simultaneously has been developed in experiments on Saccharomyces cerevisiae (commonly known as baker's or brewer's yeast). It is already known that many yeast genes perform functions similar to those of corresponding human genes; therefore, by facilitating understanding of yeast genes, the method may ultimately also contribute to the knowledge needed to treat some diseases in humans. Because of the complexity of the method and the highly specialized nature of the underlying knowledge, it is possible to give only a brief and sketchy summary here. The method involves the use of unique synthetic deoxyribonucleic acid (DNA) sequences that are denoted as DNA bar codes because of their utility as molecular labels. The method also involves the disruption of gene functions through deletion of genes. Saccharomyces cerevisiae is a particularly powerful experimental system in that multiple deletion strains easily can be pooled for parallel growth assays. Individual deletion strains recently have been created for 5,918 open reading frames, representing nearly all of the estimated 6,000 genetic loci of Saccharomyces cerevisiae. Tagging of each deletion strain with one or two unique 20-nucleotide sequences enables identification of genes affected by specific growth conditions, without prior knowledge of gene functions. Hybridization of bar-code DNA to oligonucleotide arrays can be used to measure the growth rate of each strain over several cell-division generations. The growth rate thus measured serves as an index of the fitness of the strain.

Stolc, Viktor↗

NASA NTRSDate not supplied

Analysis of Volatile Compounds from CO2 Removal Systems

One of the primary concerns when designing CO2 scrubber systems that will be integrated with a Sabatier reactor to produce water and methane is the amount of water released from the scrubber. Because the gas stream entering a Sabatier reactor must be compressed, water entering the reactor can condense and compromise the integrity of the system, thus rendering its valuable conversion capability useless. When the Johnson Space Center Environmental Chemistry Laboratory was tasked to develop an assay to quantify the water concentration in air samples from CO2 scrubbers, additional testing was also performed to see if any other compounds were being concentrated on the scrubbers. It was thought that the efficiency of the scrubber systems could be quantified by comparing the differences in samples from the ambient air on the International Space Station (ISS) to the exit gas of the scrubber. As this analysis was carried out, it became evident that the concentrations of certain volatile compounds were higher in the samples from the scrubbers than they were in nominal environmental samples. This meant these compounds were being retained and concentrated on the scrubber beds. Based on this finding, concerns were raised about their potential for these compounds to poison the Sabatier reactor. Further investigation was required to identify these compounds due to their high concentrations and unique matrix of the CO2 scrubber exhaust. This paper describes these events as well as the process that was developed to identify the volatile compounds that increased. An examination of how much the certain compounds can be concentrated by the scrubber systems is also included.

Volatiles↗

NASA NTRSDate not supplied

Analysis of Volatile Compounds From CO2 Removal Systems

Volatiles↗

NASA NTRS2010

Miniature Bioreactor System for Long-Term Cell Culture

A prototype miniature bioreactor system is designed to serve as a laboratory benchtop cell-culturing system that minimizes the need for relatively expensive equipment and reagents and can be operated under computer control, thereby reducing the time and effort required of human investigators and reducing uncertainty in results. The system includes a bioreactor, a fluid-handling subsystem, a chamber wherein the bioreactor is maintained in a controlled atmosphere at a controlled temperature, and associated control subsystems. The system can be used to culture both anchorage-dependent and suspension cells, which can be either prokaryotic or eukaryotic. Cells can be cultured for extended periods of time in this system, and samples of cells can be extracted and analyzed at specified intervals. By integrating this system with one or more microanalytical instrument(s), one can construct a complete automated analytical system that can be tailored to perform one or more of a large variety of assays.

Gonda, Steve R.↗

NASA NTRS2004

Enzyme-Cascade Analysis of the Rio Tinto Subsurface Environment: A Biosensor Experiment

The Portable Test System (PTS), designed & developed by Charles Rivers Laboratories, Inc. (Charleston, SC) is a portable instrument that was designed to perform analysis of enzymatic assays related to rapid assessment of microbial contamination (Wainwright, 2003). The enzymatic cascade of Limulus Amebocyte Lysate (LAL) is known to be one of the most sensitive techniques available for microbial detection, enabling the PTS to be evaluated as a potential life detection instrument for in situ Astrobiology missions. In the summer of 2003 the system was tested as a part of the Mars Astrobiology Research and Technology Experiment (MARTE) ground truth science campaign in the Rio Tinto Analogue environment near Nerva, Spain. The preliminary results show that the PTS analysis correlates well with the contamination control tests and the more traditional lab-based biological assays performed during the MARTE field mission. Further work will be conducted on this research during a second field campaign in 2004 and a technology demonstration of a prototype instrument that includes autonomous sample preparation will occur in 2005.

McKay, David S.↗

NASA NTRS1975

Water system virus detection

A monitoring system developed to test the capability of a water recovery system to reject the passage of viruses into the recovered water is described. A nonpathogenic marker virus, bacteriophage F2, is fed into the process stream before the recovery unit and the reclaimed water is assayed for its presence. Detection of the marker virus consists of two major components, concentration and isolation of the marker virus, and detection of the marker virus. The concentration system involves adsorption of virus to cellulose acetate filters in the presence of trivalent cations and low pH with subsequent desorption of the virus using volumes of high pH buffer. The detection of the virus is performed by a passive immune agglutination test utilizing specially prepared polystyrene particles. An engineering preliminary design was performed as a parallel effort to the laboratory development of the marker virus test system. Engineering schematics and drawings of a fully functional laboratory prototype capable of zero-G operation are presented. The instrument consists of reagent pump/metering system, reagent storage containers, a filter concentrator, an incubation/detector system, and an electronic readout and control system.

Fraser, A. S.↗

NASA NTRS2001

Evidence of root zone hypoxia in Brassica rapa L. grown in microgravity

A series of experiments was conducted aboard the U.S. space shuttle and the Mir space station to evaluate microgravity-induced root zone hypoxia in rapid-cycling Brassica (Brassica rapa L.), using both root and foliar indicators of low-oxygen stress to the root zone. Root systems from two groups of plants 15 and 30 d after planting, grown in a phenolic foam nutrient delivery system on the shuttle (STS-87), were harvested and fixed for microscopy or frozen for enzyme assays immediately postflight or following a ground-based control. Activities of fermentative enzymes were measured as indicators of root zone hypoxia and metabolism. Following 16 d of microgravity, ADH (alcohol dehydrogenase) activity was increased in the spaceflight roots 47% and 475% in the 15-d-old and 30-d-old plants, respectively, relative to the ground control. Cytochemical localization showed ADH activity in only the root tips of the space-grown plants. Shoots from plants that were grown from seed in flight in a particulate medium on the Mir station were harvested at 13 d after planting and quick-frozen and stored in flight in a gaseous nitrogen freezer or chemically fixed in flight for subsequent microscopy. When compared to material from a high-fidelity ground control, concentrations of shoot sucrose and total soluble carbohydrate were significantly greater in the spaceflight treatment according to enzymatic carbohydrate analysis. Stereological analysis of micrographs of sections from leaf and cotyledon tissue fixed in flight and compared with ground controls indicated no changes in the volume of protoplast, cell wall, and intercellular space in parenchyma cells. Within the protoplasm, the volume occupied by starch was threefold higher in the spaceflight than in the ground control, with a concomitant decrease in vacuolar volume in the spaceflight treatment. Both induction of fermentative enzyme activity in roots and accumulation of carbohydrates in foliage have been repeatedly shown to occur in response to root zone oxygen deprivation. These results indicate that root zone hypoxia is a persistent challenge in spaceflight plant growth experiments and may be caused by microgravity-induced changes in fluid and gas distribution.

long duration↗

NASA NTRS1999

Myocardialization of the cardiac outflow tract

During development, the single-circuited cardiac tube transforms into a double-circuited four-chambered heart by a complex process of remodeling, differential growth, and septation. In this process the endocardial cushion tissues of the atrioventricular junction and outflow tract (OFT) play a crucial role as they contribute to the mesenchymal components of the developing septa and valves in the developing heart. After fusion, the endocardial ridges in the proximal portion of the OFT initially form a mesenchymal outlet septum. In the adult heart, however, this outlet septum is basically a muscular structure. Hence, the mesenchyme of the proximal outlet septum has to be replaced by cardiomyocytes. We have dubbed this process "myocardialization." Our immunohistochemical analysis of staged chicken hearts demonstrates that myocardialization takes place by ingrowth of existing myocardium into the mesenchymal outlet septum. Compared to other events in cardiac septation, it is a relatively late process, being initialized around stage H/H28 and being basically completed around stage H/H38. To unravel the molecular mechanisms that are responsible for the induction and regulation of myocardialization, an in vitro culture system in which myocardialization could be mimicked and manipulated was developed. Using this in vitro myocardialization assay it was observed that under the standard culture conditions (i) whole OFT explants from stage H/H20 and younger did not spontaneously myocardialize the collagen matrix, (ii) explants from stage H/H21 and older spontaneously formed extensive myocardial networks, (iii) the myocardium of the OFT could be induced to myocardialize and was therefore "myocardialization-competent" at all stages tested (H/H16-30), (iv) myocardialization was induced by factors produced by, most likely, the nonmyocardial component of the outflow tract, (v) at none of the embryonic stages analyzed was ventricular myocardium myocardialization-competent, and finally, (vi) ventricular myocardium did not produce factors capable of supporting myocardialization. Copyright 1999 Academic Press.

NASA Discipline Cardiopulmonary↗

NASA NTRS2000

Complement activation promotes muscle inflammation during modified muscle use

Modified muscle use can result in muscle inflammation that is triggered by unidentified events. In the present investigation, we tested whether the activation of the complement system is a component of muscle inflammation that results from changes in muscle loading. Modified rat hindlimb muscle loading was achieved by removing weight-bearing from the hindlimbs for 10 days followed by reloading through normal ambulation. Experimental animals were injected with the recombinant, soluble complement receptor sCR1 to inhibit complement activation. Assays for complement C4 or factor B in sera showed that sCR1 produced large reductions in the capacity for activation of the complement system through both the classical and alternative pathways. Analysis of complement C4 concentration in serum in untreated animals showed that the classical pathway was activated during the first 2 hours of reloading. Analysis of factor B concentration in untreated animals showed activation of the alternative pathway at 6 hours of reloading. Administration of sCR1 significantly attenuated the invasion of neutrophils (-49%) and ED1(+) macrophages (-52%) that occurred in nontreated animals after 6 hours of reloading. The presence of sCR1 also reduced significantly the degree of edema by 22% as compared to untreated animals. Together, these data show that increased muscle loading activated the complement system which then briefly contributes to the early recruitment of inflammatory cells during modified muscle loading.

NASA Discipline Musculoskeletal↗

NASA NTRS2024

Flow Cytometry Methods to Monitor Immune System Dysregulation in Astronauts

Persistent immune system dysregulation has been documented to occur in astronauts participating in orbital spaceflight onboard the International Space Station. The phenomenon consists of reductions in T and NK cell function, altered cytokine profiles, persistent inflammation, and the subclinical reactivation of latent herpesviruses. In select crewmembers the dysregulation does actually lead to clinical symptoms, primarily atypical allergy or atopic dermatitis/zoster. Flow cytometry has served a central role defining the ‘immune assessment’ panel of assays that allow monitoring of astronauts. The cytometry assays which have been utilized include: 1. Peripheral leukocyte subsets 2. T cell function 3. Monocyte function 4. NK cell function 5. Intracellular cytokine analysis 6. Virus-specific T cell number (tetramer assay) 7. Virus-specific T cell function (peptide stimulation) 8. Leukocyte-bacterial challenge cytometry 9. Cytometric bead/multiplex array (soluble proteins) The use of these assays has been validated through various ISS flight investigations to define, to varying degrees, both in-flight, and post-flight immune system alterations. The kinetics of the dysregulation through the various phases of spaceflight, as well as post-flight recovery, have also been documented. To allow the science to occur within the orbital constraints of a spaceflight investigation, particular sample collection and processing techniques were developed compatible with the delays associated with terrestrial processing of in-flight samples. A subset of the assays has been adapted to routine monitoring of astronauts via a NASA ‘ISS Standard Measures’ activity, with the data from a specific crew then provided to all science investigators for that particular mission. This battery of cytometry assays has also been applied, through ground investigations, to several terrestrial ‘spaceflight analog’ populations. The purpose was to validate the analog which most closely replicates the in-flight observed pattern of alterations, generally believed to be winterover at an Antarctica station. To assist in determination of clinical risk, the assay panel has also been applied to investigations of various terrestrial patient populations, particularly zoster patients. As NASA is initiating crewed lunar missions via the ‘Artemis’ program, deployment of a miniaturized, microgravity-compatible flow cytometer, would be extremely beneficial to allow real time monitoring of crewmembers. Real time medical data could influence use of several countermeasures options during deep space missions. Several such instruments have been developed and validated to varying degrees of success. Assay details and summary findings across the various flight and ground platforms will be presented, as will current status in developing such technology for in-flight use.

Brian Crucian↗

NASA NTRS2017

Systemic Microgravity Response: Utilizing GeneLab to Develop Hypotheses for Spaceflight Risks

Biological risks associated with microgravity is a major concern for space travel. Although determination of risk has been a focus for NASA research, data examining systemic (i.e., multi- or pan-tissue) responses to space flight are sparse. The overall goal of our work is to identify potential master regulators responsible for such responses to microgravity conditions. To do this we utilized the NASA GeneLab database which contains a wide array of omics experiments, including data from: 1) different flight conditions (space shuttle (STS) missions vs. International Space Station (ISS); 2) different tissues; and 3) different types of assays that measure epigenetic, transcriptional, and protein expression changes. We have performed meta-analysis identifying potential master regulators involved with systemic responses to microgravity. The analysis used 7 different murine and rat data sets, examining the following tissues: liver, kidney, adrenal gland, thymus, mammary gland, skin, and skeletal muscle (soleus, extensor digitorum longus, tibialis anterior, quadriceps, and gastrocnemius). Using a systems biology approach, we were able to determine that p53 and immune related pathways appear central to pan-tissue microgravity responses. Evidence for a universal response in the form of consistency of change across tissues in regulatory pathways was observed in both STS and ISS experiments with varying durations; while degree of change in expression of these master regulators varied across species and strain, some change in these master regulators was universally observed. Interestingly, certain skeletal muscle (gastrocnemius and soleus) show an overall down-regulation in these genes, while in other types (extensor digitorum longus, tibialis anterior and quadriceps) they are up-regulated, suggesting certain muscle tissues may be compensating for atrophy responses caused by microgravity. Studying these organtissue-specific perturbations in molecular signaling networks, we demonstrate the value of GeneLab in characterizing potential master regulators associated with biological risks for spaceflight.

Microgravity↗

NASA NTRS2010

Automated Desalting Apparatus

Because salt and metals can mask the signature of a variety of organic molecules (like amino acids) in any given sample, an automated system to purify complex field samples has been created for the analytical techniques of electrospray ionization/ mass spectroscopy (ESI/MS), capillary electrophoresis (CE), and biological assays where unique identification requires at least some processing of complex samples. This development allows for automated sample preparation in the laboratory and analysis of complex samples in the field with multiple types of analytical instruments. Rather than using tedious, exacting protocols for desalting samples by hand, this innovation, called the Automated Sample Processing System (ASPS), takes analytes that have been extracted through high-temperature solvent extraction and introduces them into the desalting column. After 20 minutes, the eluent is produced. This clear liquid can then be directly analyzed by the techniques listed above. The current apparatus including the computer and power supplies is sturdy, has an approximate mass of 10 kg, and a volume of about 20 20 20 cm, and is undergoing further miniaturization. This system currently targets amino acids. For these molecules, a slurry of 1 g cation exchange resin in deionized water is packed into a column of the apparatus. Initial generation of the resin is done by flowing sequentially 2.3 bed volumes of 2N NaOH and 2N HCl (1 mL each) to rinse the resin, followed by .5 mL of deionized water. This makes the pH of the resin near neutral, and eliminates cross sample contamination. Afterward, 2.3 mL of extracted sample is then loaded into the column onto the top of the resin bed. Because the column is packed tightly, the sample can be applied without disturbing the resin bed. This is a vital step needed to ensure that the analytes adhere to the resin. After the sample is drained, oxalic acid (1 mL, pH 1.6-1.8, adjusted with NH4OH) is pumped into the column. Oxalic acid works as a chelating reagent to bring out metal ions, such as calcium and iron, which would otherwise interfere with amino acid analysis. After oxalic acid, 1 mL 0.01 N HCl and 1 mL deionized water is used to sequentially rinse the resin. Finally, the amino acids attached to the resin, and the analytes are eluted using 2.5 M NH4OH (1 mL), and the NH4OH eluent is collected in a vial for analysis.

Spencer, Maegan K.↗