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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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All sourcesNASA NTRSDOE OSTI
DOE OSTI2023

Natural transformation as a tool in Acinetobacter baylyi : Evolution by amplification of gene copy number

For many years, the natural competency of Acinetobacter baylyi ADP1 facilitated studies of bacterial metabolism, biochemistry, and physiology. With the advent of synthetic biology, new opportunities arise to exploit the remarkable transformability and chromosomal plasticity of this model organism. In this chapter, we describe a recently developed method, “Evolution by Amplification and Synthetic Biology” (EASy). EASy allows the targeted amplification of chromosomal segments that give rise to new phenotypes. Increased gene dosage regulates protein expression in a rudimentary fashion by establishing a chromosomal array in which copy number adjusts via recombination between repeated DNA sequences. Selective conditions enrich for cells within the population that confer a growth advantage. Under continuous selective pressure, beneficial mutations may accumulate in any genomic region. Such mutations favor decreases in the average copy number of the target region. Thus, the genetic flexibility afforded by transient copy number variation helps accelerate the selection of engineered strains with desired traits during laboratory evolution. As a result, thanks to the extremely simple genetic manipulation of Acinetobacter baylyi ADP1, the EASy method can be readily implemented by researchers without the need for advanced instrumentation or complex cloning techniques.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS1990

STS-41 Space Shuttle mission report

The STS-41 Space Shuttle Program Mission Report contains a summary of the vehicle subsystem activities on this thirty-sixth flight of the Space Shuttle and the eleventh flight of the Orbiter vehicle, Discovery (OV-103). In addition to the Discovery vehicle, the flight vehicle consisted of an External Tank (ET) (designated as ET-39/LWT-32), three Space Shuttle main engines (SSME's) (serial numbers 2011, 2031, and 2107), and two Solid Rocket Boosters (SRB's), designated as BI-040. The primary objective of the STS-41 mission was to successfully deploy the Ulysses/inertial upper stage (IUS)/payload assist module (PAM-S) spacecraft. The secondary objectives were to perform all operations necessary to support the requirements of the Shuttle Backscatter Ultraviolet (SSBUV) Spectrometer, Solid Surface Combustion Experiment (SSCE), Space Life Sciences Training Program Chromosome and Plant Cell Division in Space (CHROMEX), Voice Command System (VCS), Physiological Systems Experiment (PSE), Radiation Monitoring Experiment - 3 (RME-3), Investigations into Polymer Membrane Processing (IPMP), Air Force Maui Optical Calibration Test (AMOS), and Intelsat Solar Array Coupon (ISAC) payloads. The sequence of events for this mission is shown in tabular form. Summarized are the significant problems that occurred in the Orbiter subsystems during the mission. The official problem tracking list is presented. In addition, each Orbiter problem is cited in the subsystem discussion.

Camp, David W.↗
DOE OSTI2025

A metabolic engineering strategy for producing poly-(3-hydroxyoctanoic acid) in Escherichia coli from glycerol

Poly(3-hydroxyoctanoate) (PHO) is a medium-chain-length PHA with low crystallinity and high elongation to break ratio, unlike the brittle short-chain-PHAs like PHB. These properties make PHO a promising candidate for industrial and biomedical applications. In this paper, we demonstrated the production of PHO in Escherichia coli from a renewable and inexpensive glycerol feedstock by engineering fatty acid synthesis and β-oxidation to create a pool of 2,3-octenoyl-CoAs. In this base strain, E. coli ΔfadRABIJ, an (R)-specific enoyl-CoA hydratase (phaJ) and a PHA synthase (phaC) were expressed to produce PHO. Bioprospecting phaJ and phaC homologs from Pseudomonas aeruginosa and fadD homolog from Pseudomonas putida implicated a combination of phaJ2, phaC2, and Pp fadD genes yielded the highest PHO content from exogenously fed octanoate. Finally, when a single copy of a previously described C 8 -specific thioesterase mutant CpFatB1.2-M4-287 was integrated into the chromosome of E. coli ΔfadRABIJ, the resulting E. coli strain NHL18 was capable of producing 3.69 ± 0.146 g/L of octanoic acid. Subsequently, the integration of PHA synthesis genes in NHL18 resulting in strain SM23 allowed the cell to accumulate 15 % cell dry weight of PHO with a final titer of 1.54 ± 0.234 g/L from glycerol in fed-batch fermentation.

Biodegradable plastic↗
NASA NTRS1995

STS-68 Space Shuttle mission report

The STS-68 Space Shuttle Program Mission Report summarizes the Payload activities as well as the Orbiter, External Tank (ET), Solid Rocket Booster (SRB), Redesigned Solid Rocket Motor (RSRM), and the Space Shuttle main engine (SSME) systems performance during the sixty-fifth flight of the Space Shuttle Program and the seventh flight of the Orbiter vehicle Endeavour (OV-105). In addition to the Orbiter, the flight vehicle consisted of an ET that was designated ET-65; three SSMEs that were designated as serial numbers 2028, 2033, and 2026 in positions 1, 2, and 3, respectively; and two SRBs that were designated BI-067. The RSRMs that were installed in each SRB were designated as 360W040A for the left SRB and 360W040B for the right SRB. The primary objective of this flight was to successfully perform the operations of the Space Radar Laboratory-2 (SRL-2). The secondary objectives of the flight were to perform the operations of the Chromosome and Plant Cell Division in Space (CHROMEX), the Commercial Protein Crystal Growth (CPCG), the Biological Research in Canisters (BRIC), the Cosmic Radiation Effects and Activation Monitor (CREAM), the Military Application of Ship Tracks (MAST), and five Get-Away Special (GAS) payloads.

Fricke, Robert W., Jr.↗
NASA NTRS1993

STS-54 Space Shuttle mission report

The STS-54 Space Shuttle Program Mission Report is a summary of the Orbiter, External Tank (ET), Solid Rocket Booster/Redesigned Solid Rocket Motor (SRB/RSRM), and the Space Shuttle Main Engine (SSME) subsystems performance during this fifty-third flight of the Space Shuttle Program, and the third flight of the Orbiter vehicle Endeavour (OV-105). In addition to the Orbiter, the flight vehicle consisted of an ET, which was designated ET-51; three SSME's, which were serial numbers 2019, 2033, and 2018 in positions 1, 2, and 3, respectively; and two retrievable and reusable SRB's which were designated BI-056. The lightweight RSRM's that were installed in each SRB were designated 360L029A for the left SRB, and 360L029B for the right SRB. The primary objectives of this flight were to perform the operations to deploy the Tracking and Data Relay Satellite-F/Inertial Upper Stage payload and to fulfill the requirements of the Diffuse X-Ray Spectrometer (DXS) payload. The secondary objective was to fly the Chromosome and Plant Cell Division in Space (CHROMEX), Commercial Generic Bioprocessing Apparatus (CGBA), Physiological and Anatomical Rodent Experiment (PARE), and the Solid Surface Combustion Experiment (SSCE). In addition to presenting a summary of subsystem performance, this report also discusses each Orbiter, ET, SSME, SRB, and RSRM in-flight anomaly in the applicable section of the report. The official tracking number for each in-flight anomaly, assigned by the cognizant project, is also shown. All times are given in Greenwich mean time (G.m.t.) and mission elapsed time (MET).

Fricke, Robert W., Jr.↗
DOE OSTI2022

Artificial intelligence based analysis of nanoindentation load–displacement data using a genetic algorithm

In this work, we developed an automated tool, Nanoindentation Neo package for the analysis of nanoindentation load–displacement curves using a Genetic Algorithm (GA) applied to the Oliver-Pharr method (Oliver et al.,1992). For some materials, such as polycrystalline isotropic graphites, Least Squares Fitting (LSF) of the unload curve can produce unrealistic fit parameters. These graphites exhibit sharply peaked unloading curves not easily fit using the LSF, which tends to overestimate the indenter tip geometry parameter. To tackle this problem, we extended our general materials characterization tool Neo for EXAFS analysis (Terry et al., 2021) to fit nanoindentation data. Nanoindentation Neo automatically processes and analyzes nanoindentation data with minimal user input while producing meaningful fit parameters. GA, a robust metaheuristic method, begins with a population of temporary solutions using model parameters called chromosomes; from these we evaluate a fitness value for each solution, and select the best solutions to mix with random solutions producing the next generation. A mutation operator then modifies existing solutions by random perturbations, and the optimal solution is selected. We tested the GA method using Silica and Al reference standards. We fit samples of graphite and a high entropy alloy (HEA) consisting of BCC and FCC phases.

42 ENGINEERING↗
DOE OSTI2023

Clonality, local population structure, and gametophyte sex ratios in cryptic species of the Sphagnum magellanicum complex

Sphagnum (peatmoss) comprises a moss (Bryophyta) clade with approximately 300-500 species. The genus has unparalleled ecological importance because Sphagnum-dominated peatlands store almost a third of the terrestrial carbon pool and peatmosses engineer the formation and microtopography of peatlands. Genomic resources for Sphagnum are being actively expanded, but many aspects of their biology are still poorly known. Among these are the degree to which Sphagnum species reproduce asexually, and the relative frequencies of male and female gametophytes in these haploid-dominant plants. Here, we assess clonality and gametophyte sex ratios and test hypotheses about the local-scale distribution of clones and sexes in four North American species of the S. magellanicum complex. These four species are difficult to distinguish morphologically and are very closely related. We also assess microbial communities associated with Sphagnum host plant clones and sexes at two sites. 405 samples of the four species, representing 57 populations, were subjected to RADseq. Analyses of population structure and clonality based on the molecular data utilized both phylogenetic and phenetic approaches. Multi-locus genotypes (genets) were identified using the RADseq data. Sexes of sampled ramets were determined using a molecular approach that utilized coverage of loci on the sex chromosomes after the method was validated using a sample of plants that expressed sex phenotypically. Sex ratios were estimated for each species, and populations within species. Differences in fitness between genets was estimated as the numbers of ramets each genet comprised. Degree of clonality (numbers of genets/numbers of ramets [samples]) within species, among sites, and between gametophyte sexes were estimated. Sex ratios were estimated for each species, and populations within species. Sphagnum-associated microbial communities were assessed at two sites in relation to Sphagnum clonality and sex. All four species appear to engage in a mixture of sexual and asexual (clonal) reproduction. A single ramet represents most genets but 2-8 ramets were detected for some genets. Only one genet is represented by ramets in multiple populations; all other genets are restricted to a single population. Within populations ramets of individual genets are spatially clustered, suggesting limited dispersal even within peatlands. Sex ratios are male-biased in S. diabolicum but female-biased in the other three species, although significantly so only in S. divinum. Neither species nor males/females differ in levels of clonal propagation. At St. Regis Lake (NY) and Franklin Bog (VT), microbial community composition is strongly differentiated between the sites, but differences between species, genets, and sexes were not detected. Within S. divinum, however, female gametophytes harbored 2-3 times the number oi of microbial taxa as males. These four Sphagnum species all exhibit a similar reproductive patterns that result from a mixture of sexual and asexual reproduction. The spatial patterns of clonally replicated ramets of genets suggest that these species fall between the so-called phalanx patterns where genets abut one another but do not extensively mix, because of limited ramet fragmentation, and the guerrilla patterns where extensive genet fragmentation and dispersal results in greater mixing of different genets. Although sex ratios in bryophytes are most often female-biased, both male and female biases occur in this complex of closely related species. The association of far greater microbial diversity for female gametophytes in S. divinum, which has a female-biased sex ratio, suggests additional research to determine if levels of microbial diversity are consistently correlated with differing patterns of sex ratio biases.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS2015

Fall 2015 NASA Internship, and Space Radiation Health Project

This fall, I was fortunate enough to have been able to participate in an internship at NASA's Lyndon B. Johnson Space Center. I was placed into the Human Health & Performance Directorate, where I was specifically tasked to work with Dr. Zarana Patel, researching the impacts of cosmic level radiation on human cells. Using different laboratory techniques, we were able to examine the cells to see if any damage had been done due to radiation exposure, and if so, how much damage was done. Cell culture samples were exposed at different doses, and fixed at different time points so that we could accumulate a large pool of quantifiable data. After examining quantifiable results relative to the impacts of space radiation on the human body at the cellular and chromosomal level, researchers can defer to different areas of the space program that have to do with astronaut safety, and research and development (extravehicular mobility unit construction, vehicle design and construction, etc.). This experience has been very eye-opening, and I was able to learn quite a bit. I learned some new laboratory techniques, and I did my best to try and learn new ways to balance such a hectic work and school schedule. I also learned some very intimate thing about working at NASA; I learned that far more people want to watch you succeed, rather than watch you fail, and I also learned that this is a place that is alive with innovators and explorers - people who have a sole purpose of exploring space for the betterment of humanity, and not for any other reason. It's truly inspiring. All of these experiences during my internship have impacted me in a really profound way, so much that my educational and career goals are completely different than when I started. I started out as a biotechnology major, and I discovered recently toward the end of the internship, that I don't want to work in a lab, nor was I as enthralled by biological life sciences as a believed myself to be. Taking that all into consideration, I've actually changed my major to mechanical engineering. I discovered that I enjoy building things, and I enjoy learning about materials and interactions between different things. And I quickly became obsessed with rocket and aerospace engineering, so I've decided that after a mechanical engineering degree, I will be pursuing an advanced degree in aerospace engineering. One final way that I was effected by this internship, is that I discovered that I don't want to have a career at NASA. I love this agency with all of my heart, but I refuse to allow my innovation to be bound by a scientifically illiterate congress. As such, I have decided to pursue commercial aerospace companies, such as Space, XCOR, Masten Space Systems, Orbital ATK, and many, many, more. Maybe one day I'll end up back here. I believe in what this agency is doing with my whole heart, and it's unfortunate to see them curtailed in some capacities as a result of budgetary constraints, brought on by people who don't fully understand the effort behind putting people in to space. All in all, this experience has been the best experience of my life - literally a childhood dream came true during this experience - and I cannot adequately explain how grateful I am to have been here for the past sixteen weeks.

Patience, Luke↗
NASA NTRS1999

Three-Dimensional, Transgenic Cell Models to Quantify Space Genotoxic Effects

The space environment contains radiation and chemical agents known to be mutagenic and carcinogenic to humans. Additionally, microgravity is a complicating factor that may modify or synergize induced genotoxic effects. Most in vitro models fail to use human cells (making risk extrapolation to humans more difficult), overlook the dynamic effect of tissue intercellular interactions on genotoxic damage, and lack the sensitivity required to measure low-dose effects. Currently a need exists for a model test system that simulates cellular interactions present in tissue, and can be used to quantify genotoxic damage induced by low levels of radiation and chemicals, and extrapolate assessed risk to humans. A state-of-the-art, three-dimensional, multicellular tissue equivalent cell culture model will be presented. It consists of mammalian cells genetically engineered to contain multiple copies of defined target genes for genotoxic assessment,. NASA-designed bioreactors were used to coculture mammalian cells into spheroids, The cells used were human mammary epithelial cells (H184135) and Stratagene's (Austin, Texas) Big Blue(TM) Rat 2 lambda fibroblasts. The fibroblasts were genetically engineered to contain -a high-density target gene for mutagenesis (60 copies of lacl/LacZ per cell). Tissue equivalent spheroids were routinely produced by inoculation of 2 to 7 X 10(exp 5) fibroblasts with Cytodex 3 beads (150 micrometers in diameter). at a 20:1 cell:bead ratio, into 50-ml HARV bioreactors (Synthecon, Inc.). Fibroblasts were cultured for 5 days, an equivalent number of epithelial cells added, and the fibroblast/epithelial cell coculture continued for 21 days. Three-dimensional spheroids with diameters ranging from 400 to 600 micrometers were obtained. Histological and immunohistochemical Characterization revealed i) both cell types present in the spheroids, with fibroblasts located primarily in the center, surrounded by epithelial cells; ii) synthesis of extracellular matrix; and iii,, mitotic cells located throughout the spheroids. Spheroidal integrity and cell viability were retained for the 30-day test period after removal of spheroids from the bioreactor. Potential utility of this three-dimensional, transgenic model for genotoxicity was initially assessed by exposure of spheroids to 0-2 Gy neon at dose rates of 0.3 to 1.5 Gy/min (National Institute of Radiological Sciences, Chiba, Japan). Quantification of mutation at the lacl gene revealed a linear dose response for mutation induction. Limited sequencing analysis of mutant clones revealed higher frequencies of deletions and multiple base sequence changes with increasing dose. These results suggest that our three-dimensional, transgenic model is applicable to a wide variety of studies involving the quantification, identification, and characterization of genotoxicity incurred in space and on Earth. This model uniquely allows investigation of the interaction of relevant factors, namely cell-to-cell interactions and the mechanistic interaction of microgravity with radiation insults and DNA repair. Using this three-dimensional model will allow us to obtain dual genotoxic information (i.e., mutation rate plus chromosome aberration data) from the same system so that one endpoint can be used to reference the other, thereby increasing the fidelity of the data set. Moreover, the tissue-equivalent nature of the three-dimensional model provides high confidence for relevance of risk assessment, i.e., the establishment of quality factors directly applicable to the microgravity environment.

Gonda, S. R.↗
DOE OSTI2023

Molecular and biochemical characterization of rice developed through conventional integration of nDart1-0 transposon gene

Mutations, the genetic variations in genomic sequences, play an important role in molecular biology and biotechnology. During DNA replication or meiosis, one of the mutations is transposons or jumping genes. An indigenous transposon nDart1-0 was successfully introduced into local indica cultivar Basmati-370 from transposon-tagged line viz., GR-7895 (japonica genotype) through conventional breeding technique, successive backcrossing. Plants from segregating populationsshowed variegated phenotypes were tagged as BM-37 mutants. Blast analysis of the sequence data revealed that the GTP-binding protein, located on the BAC clone OJ1781_H11 of chromosome 5, contained an insertion of DNA transposon nDart1-0. The nDart1-0 has “A” at position 254 bp, whereas nDart1 homologs have “G”, which efficiently distinguishes nDart1-0 from its homologs. The histological analysis revealed that the chloroplast of mesophyll cells in BM-37 was disrupted with reduction in size of starch granules and higher number of osmophillic plastoglobuli, which resulted in decreased chlorophyll contents and carotenoids, gas exchange parameters (Pn, g, E, Ci), and reduced expression level of genes associated with chlorophyll biosynthesis, photosynthesis and chloroplast development. Along with the rise of GTP protein, the salicylic acid (SA) and gibberellic acid (GA) and antioxidant contents(SOD) and MDA levels significantly enhanced, while, the cytokinins (CK), ascorbate peroxidase (APX), catalase (CAT), total flavanoid contents (TFC) and total phenolic contents (TPC) significantly reduced in BM-37 mutant plants as compared with WT plants. These results support the notion that GTP-binding proteins influence the process underlying chloroplast formation. Therefore, it is anticipated that to combat biotic or abiotic stress conditions, the nDart1-0 tagged mutant (BM-37) of Basmati-370 would be beneficial.

60 APPLIED LIFE SCIENCES↗