Development of analytical techniques for hydrocarbons in mineral aggregates
Assay of organic chemical constituents including hydrocarbons of terrestrial sediments varying in age from recent ocean bottom sediments to meteorite samples
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Assay of organic chemical constituents including hydrocarbons of terrestrial sediments varying in age from recent ocean bottom sediments to meteorite samples
The Mars Exploration Rover (MER) "Opportunity" has explored chemically-enriched sedimentary outcrops at Meridiani Planum, Mars. In its first year, three different crater sites - Eagle, Fram and Endurance - have been explored. Nineteen high-interest outcrop rocks were investigated by first grinding a hole to reach the interior (using the Rock Abrasion Tool, RAT), and then conducting APXS (alpha particle x-ray spectrometry) analysis, MB (M ssbauer) analysis, and close up imaging (MI, microscopic imager). Sixteen elements and four Fe-bearing minerals were assayed to good accuracy in each sample, producing 380 compositional data points. The Miniature Thermal Emission Spectrometer (Mini- TES) obtained spectra on outcrop materials which provide direct indication of several mineral classes. Preliminary reports on Eagle crater and three RAT samples have been published. Chemical trends and a derived mineralogical model for all RAT d outcrop samples to date has been developed.
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Measurements and analyses of resources within permanently shadowed craters and along steep embankments on extraterrestrial surfaces pose extreme challenges since astronauts and rovers cannot access them. However, these areas offer the potential for the greatest scientific returns. Researchers at the NASA Langley Research Center (LaRC) invented a deployable wireless micro-spectrometer that can access those challenging areas and deliver telemetry data to a receiver safely outside those extreme environments. The micro-spectrometers are designed for deployment by an astronaut, a rover, or a lander flying overhead. This device can also be installed on rover tires, under the astronaut’s shoes, or on a cane stick. The bullet-like, expendable micro-spectrometer can penetrate into soil to spectrally identify the components of soil, such as water, He-3, or other minerals. The signals from the soil assay data are transmitted to a mother station through a telemetry system. This LaRC developed micro-spectrometer bullet consists of micro-spectrometer optics with an all-imbedded, burst-mode, light-emitting-diode ultraviolet (LED UV) light source, a super-capacitor with control electronics, and telemetry electronics. Prototypes have been fabricated to demonstrate a spectral assay of soil components. Further maturation of this technology would be necessary for demonstrations on the Moon.
Scientific measurements within craters and along steep embankments on Mars pose extreme challenges since astronauts and rovers cannot access them. However, these areas offer the potential for thegreatest scientific return. NASA Langley Research Center (LaRC) has invented a deployable wireless micro-spectrometer [1-13] capable of accessing thosechallenging areas and, using telemetry, send measurements to a receiver safely outside those extreme environments. A prototype of the micro-spectrometer concept was fabricated and successfullytested for measurement of dopamine as a neural probe [14]. The bullet-sized micro-spectrometer concept can be designed for deployment by an astronaut, a rover, an aerial drone, or a lander, and the measurements broadcasted back to a receiver [13,15,16]. This device can also be installed on rovertires, under the astronaut’s boots, or a cane stick. The bullet-like consumable micro-spectrometer can penetrate soil to spectrally identify the components ofthe soil, such as water or minerals. The signals of the soil assay data are transmitted to a mother station through a telemetry system. The LaRC developed micro-spectrometer bullet consists of micro-spectrometer optics with an all imbedded burst-mode LED UV light source, a super-capacitor with control electronics, and telemetry electronics. Prototypes have been fabricated to demonstrate spectral assay of soil components. Further maturation of this technology would be necessary for demonstration on Mars. Future work includes a potential R&D program that includes other NASA Centers and industry partners interested in developing a business case that requires knowledge of the locations of extraterrestrial ice, for instance.
Plants strongly influence soil properties through rhizodeposition, in which exudates diffuse from roots, additional secretions are actively released, and root cells are sloughed into the soil. This contribution by plants of carbon compounds belowground is at the core of soil health, water holding capacity, and the soil carbon storage that pulls carbon dioxide out of the atmosphere. Once in soil, organic matter can bind with minerals such as iron hydroxides, where it can be protected from microbial attack for millenia, preserving very large terrestrial soil carbon pools. However, those same compounds contributed by roots to soil may also destabilize the long-term protective associations of SOM with minerals, making that soil organic matter (SOM) more vulnerable to microbial attack and decomposition. Plant roots thus influence both the buildup and breakdown of soil carbon pools. DOE’s E3SM Land Model (ELM) includes a representation of soil carbon storage on minerals, but the potential vulnerability of SOM–mineral associations to effects of rhizodeposition is not yet represented in ELM. To begin testing for this effect of rhizodeposition on soil carbon storage and decomposition, we worked to develop a novel approach during this TES Exploratory project DE-SC0019142 – we harnessed the power of plant viral infection. We examined whether plant virus infection can serve as a tool to intensify rhizodeposition at the root surface, and therefore possibly intensify mobilization of SOM from minerals making it visible to our analytical techniques. Viral infection is widespread in terrestrial ecosystems; 25-70% of plants have virus infection, yet the influence of such infection on root traits and terrestrial soil carbon dynamics remains largely unexplored. We used two plant hosts: the annual Avena sativa (oats) and the genetically tractable, model grass Brachypodium distachyon. These grasses were infected with the broad host range virus Barley Yellow Dwarf Virus (BYDV) via aphids (Rhopalosiphum padi). BYDV infects at least 150 grass species in agricultural and natural ecosystems, and in previous experiments, oats infected with BYDV had roots that were very sticky to the touch, strongly suggesting that infection altered rhizodeposition. We developed this new experimental approach mostly in a one virus (Barley Yellow Dwarf Virus)–one plant (Avena sativa) system. (Several effects of infection in a Brachypodium-BYDV system were similar in nature to effects on Avena sativa, but were more variable.) In the BYDV-Avena system, we developed protocols for consistently infecting target plants (and avoiding infection of control plants) using aphid caging on leaves. We measured that infected plants exhibited reduced photosynthesis, plant (including root) biomass, and root:shoot ratio, as well as simplified root system architecture. We established procedures for sampling the organic compounds carried specifically in phloem (vascular tissue) of leaves and roots, using aphid stylectomy. We used FTICR-MS, Orbitrap GC-MS, and LC-MS/MS to analyze organic compounds in phloem, liquid around roots of plants grown hydroponically, and pore water around roots in soil, and found differences in the compounds in solution bathing roots when infected and uninfected plants were grown hydroponically. Finally, we synthesized isotopically-labeled mineral–organic matter (MAOM) associations in the lab and developed assays using them in solution and in soil. Assays quantified the extent and rate of mineralization of labeled MAOM that was mobilized by functionally distinct rhizodeposits and then attacked by microbes. Two mechanisms for MAOM mobilization emerged, with distinct dynamics. During “direct” mobilization, rhizodeposits such as the strong ligand oxalic acid could drive rapid dissolution of minerals, mobilizing MAOM. During “indirect” mobilization, rhizodeposits such as the simple sugar glucose did not attack minerals directly but instead intensified microbial activity, which led to mobilization via changes in e.g. pH, Eh, and microbial metabolite production (Li et al. 2021). Mechanistic understanding derived from these data and our ongoing experiments using these techniques will inform future development of ELM. Plant roots not only contribute newly fixed organic compounds to soils, but also root activities can drive mineralization of the carbon and nutrients mobilized off minerals via “indirect” or “direct” mechanisms. Using viral infection as a new tool, ongoing combined experimentation and modeling will explore the strength and larger-scale significance of the cascade of processes from rhizodeposition to MAOM mobilization for soil carbon storage and nutrient cycling in terrestrial ecosystems. And if viral infection leads quite generally to “sticky roots”, our perception of the potential importance of prevalent virus infection in terrestrial landscapes will be transformed.
Broad tissue degeneration and the failure of normal tissue regenerative processes in microgravity because of mechanical unloading are increasing concerns for sustaining life in space as the duration of future flight missions increases. Work in our laboratory has identified normal adult stem cell-based tissue regenerative processes, such as the formation of new bone, cartilage, and immune cells, as being particularly sensitive to the stresses of mechanical unloading in microgravity. Our studies have also identified the inhibition of differentiation of marrow mesenchymal stem cells and activation of CDKN1ap21-mediated cell cycle arrest in proliferative osteoprecursor cells on the bone surface as potential mechanisms for spaceflight-induced skeletal changes. This finding, in combination with the role of CDKN1ap21 as a suppressor of mammalian tissue regeneration, suggests that this gene could be responsible for suppressing stem cell-based tissue regeneration in response to disuse. In this work, we hypothesized that CDKN1ap21 regulates regenerative bone formation in response to alterations in mechanical load and tested this hypothesis by studying the skeletal phenotype and stem cell regenerative ability of juvenile (4-11 weeks old) and adult (18 weeks-12 months old) p21 (--) knockout (KO) mice. Additionally, we analyzed bone micro-architectural properties, bone formation rates and differentiation capacity of bone marrow stem cells (BMSCs) from male and female KO mice exposed to hindlimb unloading (HU) for 15-30 days. We found that juvenile KO mice exhibited increased femoral trabecular and cortical bone formation, whilst three-point bending of the tibias from KO mice showed decreased bone stiffness. Conversely, adult KO mice exhibited no significant differences in micro-architectural properties compared to WT (wild-type) but woven bone structure was indicative of rapid bone remodeling. Furthermore, cortical bone properties showed similar characteristics to aged bone, including increased cross-sectional area and perimeter, whilst three-point bending showed increased stiffness and toughness. Interestingly, in-vitro, KO mice exhibited increased differentiation and mineralized nodule formation in osteoblastogenesis assays compared to WT. Preliminary results from CDKN1ap21 KO mice subjected to HU suggest altered sensitivity to mechanical unloading resulting in decreased cortical thickness compared to WT mice. However, KO mice subjected to short and long-duration HU show increased in-vitro differentiation potential of BMSCs to from form mature, mineral-forming osteoblasts, indicating maintenance of regenerative potential. Analysis of bone formation rates, cell proliferation rates and key genes of interest are currently underway. These results indicate a novel role for CDKN1ap21 in load-dependent osteoprogenitor proliferation and differentiation and that deletion of CDKN1ap21 results in an age-dependent release of osteoblast proliferation inhibition and increased bone formation and turnover.
The Modular Autonomous Research System (MARS) was developed to address the pressing need for faster, more reliable, and more adaptable scientific discovery. Traditional experimentation is limited by manual labor, long cycle times, and fragmented data streams, which constrain the ability to explore complex chemical and materials design spaces. To overcome these limitations, we created an integrated, modular platform that combines laboratory robotics, diverse measurement instruments, and a central data infrastructure with artificial intelligence–driven decision-making. The system links liquid handling robots, robotic arms, and optical plate readers into a closed loop where experiments are executed automatically, data is analyzed in real time, and subsequent experimental conditions are adaptively chosen to maximize information gain. Over the course of the project, MARS was validated on two primary test cases—spectroscopic metal–ligand binding assays and peptide-directed mineralization—which highlighted the system’s ability to handle uncertainty and variability in experimental measurements. To further demonstrate modularity and extensibility, we also established additional testbeds in electrochemistry for catalyst discovery and electrolyte formulation for advanced batteries. The results show that MARS can reliably conduct autonomous campaigns with minimal human intervention, adapt to distinct scientific domains, and provide a scalable model for future self-driving laboratories. This work establishes new capabilities for modular, uncertainty-aware automation and directly supports the need for advanced, data-driven research platforms capable of accelerating discovery across a wide range of scientific and national security missions.
The author has identified the following significant results. Major structural trends have been compiled on five separate maps, at a scale of 1:1,000,000, of Utah and Nevada from ERTS-1 imagery. An arbitrary length of ten kilometers has been chosen as a minimum length of the trends. The selection is based upon: (1) obvious displacement of structures; (2) continuity or persistence of trends across structures; (3) line-up of outcrop patterns, drainage, erosional features or vegetation; and (4) near-linear trends. Several recognizable trend directions have noted, viz., N 10-15 deg W, N 35 deg W, N 80 deg W, N 30 deg E, and E-W. More than 1500 structural trends have been identified, some of which are mineralized and extend into pediment or shallow alluvial cover. Those fracture trends that exhibit mineralization in exposed bedrock will be assayed for mercury content with a soil-gas analyzer and similar collection will be done over alluvial posture blankets by the same technique with the hope of discovering blind mineralized zones that are not exposed on the surface.
Recent studies on the interactive effects of hypobaria, low temperatures, and CO2-enriched anoxic atmospheres on the growth of 37 species of mesophilic bacteria identified 14 potential biocidal agents that might affect microbial survival and growth on the martian surface. Biocidal or inhibitory factors include (not in priority): (1) solar UV irradiation, (2) low pressure, (3) extreme desiccating conditions, (4) extreme diurnal temperature fluctuations, (5) solar particle events, (6) galactic cosmic rays, (7) UV-glow discharge from blowing dust, (8) solar UV-induced volatile oxidants [e.g., O2(-), O(-), H2O2, O3], (9) globally distributed oxidizing soils, (10) extremely high salts levels [e.g., MgCl2, NaCl, FeSO4, and MgSO4] in surficial soils at some sites on Mars, (11) high concentrations of heavy metals in martian soils, (12) likely acidic conditions in martian fines, (13) high CO2 concentrations in the global atmosphere, and (14) perchlorate-rich soils. Despite these extreme conditions several studies have demonstrated that dormant spores or vegetative cells of terrestrial microorganisms can survive simulated martian conditions as long as they are protected from UV irradiation. What has not been explored in depth are the effects of potential biotoxic geochemical components of the martian regolith on the survival and growth of microorganisms. The primary objectives of the research included: (1) prepare and characterize Mars analog soils amended with potential biotoxic levels of sulfates, salts, acidifying minerals, etc.; and (2) use the simulants to conduct biotoxicity assays to determine if terrestrial microorganisms from spacecraft can survive direct exposure to the analog soils.
Recent evidence from the Opportunity and Spirit rovers suggests that the soils on Mars might be very high in biotoxic materials induding sulfate salts, chlorides, and acidifying agents. Yet, very little is known about how the chemistries of Mars soils might affect the survival and growth of terrestrial microorganisms. The primary objectives of the proposed research will be to: (1) prepare and characterize Mars analog soils amended with potential biotoxic levels of sulfates, chlorides, and acidifying minerals; (2) use the stimulants to conduct a series of toxicology assays to determine if terrestrial microorganisms from spacecraft or extreme environments can survive direct exposure to the biotoxic soils, and (3) mix soils from extreme environments on Earth into Mars analog soils to determine if terrestrial microorganisms can grow and replicate under Martian conditions. The Mars analog soils will be thoroughly characterized by a wide diversity of soil chemistry assays to determine the exact nature of the soluble biotoxic components following hydration. The microbial experiments will be designed to test the effects of Mars stimulants on microbial survival, growth and replication during direct challenge experiments. Toxicology experiments will be designed to mimic terrestrial microbes coming into contact with biotoxic soils with and without liquid water. Results are expected to help" ... characterize the limits of life in ... planetary environments ... " and may help constrain the search for life on Mars.
Recent evidence from the Opportunity and Spirit rovers and the Mars Express mission suggests that the soils on Mars might be very high in biotoxic materials including sulfate salts, chlorides, and acidifying agents. Yet, very little is known about how the chemistries of Mars soils might affect the survival and growth of terrestrial microorganisms. The primary objectives of the research included: (1) prepare and characterize Mars analog soils amended with potential biotoxic levels of sulfates, chlorides, and acidifying minerals; and (2) use the simulants to conduct a series of toxicology assays to determine if terrestrial microorganisms from spacecraft can survive direct exposure to the biotoxic soils.
A nuclear morphometric assay for preosteoblasts is introduced as a cell-kinetic technique, applicable to routine histological preparations of mineralized tissue. Because this method is a morphological marker for osteoblast precursor cell differentiation, it provides a new dimension for determining the mechanism of osteoblast histogenesis. Osteoblast precursors of the periodontal ligament are a mixed population of progenitors, kinetically separable into two distinct groups according to nuclear size. Preosteoblasts, the immediate proliferating precursors of osteoblasts, have large nuclei (greater than 170 micrometers3) and are derived from relatively undifferentiated fibroblastlike cells, which have smaller nuclei (less than 80 micrometers3). Increase in nuclear volume, during G1 phase of the cell cycle, is apparently a morphological manifestation of change in genomic expression. This key event in preosteoblast differentiation is related to mechanical stress/strain and may be an important rate-limiting step in osteoblast histogenesis.
In an attempt to verify the results reported previously concerning the phytotoxicity of wet-oxidation (wet-ox) products, lettuce solids were suspended in water and then heated to 548 K for 3.6 ksec (1 hr) under 4.1x10 to the -7 Pa (400 psig at 294 K) oxygen pressure and 1.52x10 to the 8th (1500 psig at 548 K) total pressure. Such treatment resulted in oxidation of 80% of the initial organic carbon to carbon dioxide. Thirty-three percent of the remaining organic carbon was present in acetic acid. Organic nitrogen in the feed was decreased 90% by the wet-ox treatment. Ammonia and nitrogen gas were the main nitrogen products. Analysis of the liquid product of wet-ox indicated that most of the minerals essential for plant growth were present. However, when tested using a lettuce-root growth-rate assay, the solution was toxic. This toxicity was not due to excessive salt or ammonia or to an improper pH. Analysis of the wet-ox solution revealed the presence of silver and chromium, thus implicating reactor corrosion as the cause of the phytotoxicity. Both cation and anion exchange resins removed the silver and the toxicity of the liquid effluent, indicating silver as the toxic component. Uptake of both silver and chromium by lettuce roots correlated with diminished root growth. Toxicity of the solution from wet-ox was not observed when precautions were taken to minimize contact of the liquid in the reactor with the metal reactor components.
Remote sensor data (primarily LANDSAT) was analyzed by photogeologic and computer-assisted enhancement techniques to evaluate the metallic mineral potential of Baja California. Overlays were prepared at 1:1,000,000 and 1:500,000 and included known geologic relationships and mineral occurrences, lineament, drainage and structural patterns, tonal anomalies, and enhancement results. Computer-assisted enhancement and classification of the test sites was performed using the IMAGE 100 system to identify subtle tonal anomalies thought related to mineralization using known sites as analysis guides. Mineral potential maps of Baja California were generated from these analyses and the ten highest priority targets visited. Preliminary assay results (atomic absorption analysis) for the samples recovered showed moderate to high geochemical anomalies for Copper (10 of 12 samples), Zinc (3 of 12 samples) and Lead (4 of 12 samples).
Neonatal rat calvarial osteoblasts were cultured in 90% porous, 75:25 poly(DL-lactic-co-glycolic acid) (PLGA) foam scaffolds for up to 56 days to examine the effects of the cell seeding density, scaffold pore size, and foam thickness on the proliferation and function of the cells in this three-dimensional environment. Osteoblasts were seeded at either 11.1 x 10(5) or 22.1 x 10(5) cells per cm2 onto PLGA scaffolds having pore sizes in the range of 150-300 or 500-710 microm with a thickness of either 1.9 or 3.2 mm. After 1 day in culture, 75.6 and 68.6% of the seeded cells attached and proliferated on the 1.9 mm thick scaffolds of 150-300 microm pore size for the low and high seeding densities, respectively. The number of osteoblasts continued to increase throughout the study and eventually leveled off near 56 days, as indicated by a quantitative DNA assay. Osteoblast/foam constructs with a low cell seeding density achieved comparable DNA content and alkaline phosphatase (ALPase) activity after 14 days, and mineralization results after 56 days to those with a high cell seeding density. A maximum penetration depth of osseous tissue of 220+/-40 microm was reached after 56 days in the osteoblast/foam constructs of 150-300 microm pore size initially seeded with a high cell density. For constructs of 500-710 microm pore size, the penetration depth was 190+/-40 microm under the same conditions. Scaffold pore size and thickness did not significantly affect the proliferation or function of osteoblasts as demonstrated by DNA content, ALPase activity, and mineralized tissue formation. These data show that comparable bone-like tissues can be engineered in vitro over a 56 day period using different rat calvarial osteoblast seeding densities onto biodegradable polymer scaffolds with pore sizes in the range of 150-710 microm. When compared with the results of a previous study where similar polymer scaffolds were seeded and cultured with marrow stromal cells, this study demonstrates that PLGA foams are suitable substrates for osteoblast growth and differentiated function independent of cell source.
Understanding the response of bone to mechanical loading (unloading) is extremely important in defining the means of adaptation of the body to a variety of environmental conditions such as during heightened physical activity or in extended explorations of space or the sea floor. The mechanisms of the adaptive response of bone are not well defined, but undoubtedly they involve changes occurring at the cellular level of bone structure. This proposal has intended to examine the hypothesis that the loading (unloading) response of bone is mediated by specific cells through modifications of their activity cytoskeletal elements, and/or elaboration of their extracellular matrices. For this purpose, this laboratory has utilized the results of a number of previous studies defining molecular biological, biochemical, morphological, and ultrastructural events of the reproducible mineralization of a primary bone cell (osteoblast) culture system under normal loading (1G gravity level). These data and the culture system then were examined following the use of the cultures in two NASA shuttle flights, STS-59 and STS-63. The cells collected from each of the flights were compared to respective synchronous ground (1G) control cells examined as the flight samples were simultaneously analyzed and to other control cells maintained at 1G until the time of shuttle launch, at which point they were terminated and studied (defined as basal cells). Each of the cell cultures was assayed in terms of metabolic markers- gene expression; synthesis and secretion of collagen and non-collagenous proteins, including certain cytoskeletal components; assembly of collagen into macrostructural arrays- formation of mineral; and interaction of collagen and mineral crystals during calcification of the cultures. The work has utilized a combination of biochemical techniques (radiolabeling, electrophoresis, fluorography, Western and Northern Blotting, and light microscopic immunofluorescence) and structural methods (conventional and high voltage electron microscopy, inununocytochemistry, stereomicroscopy, and 3D image reconstruction). The studies have provided new knowledge of aspects of bone cell development and structural regulation, extracellular matrix assembly, and mineralization during spaceflight and under normal gravity. The information has contributed to insights into the means in general by which cells respond and adapt to different conditions of gravity (loading). The data may as well have suggested an underlying basis for the observed loss of bone by vertebrates, including man, in microgravity; and these scientific results may have implications for understanding bone loss following fracture healing and extended periods of inactivity such as during long-term bedrest.