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NASA NTRS1985

Distribution and abundance of organic thiols

The role of glutathione (GSH) in protecting against the toxicity of oxygen and oxygen by products is well established for all eukaryotes studied except Entamoeba histolytica which lacks mitochrondria, chloroplasts, and microtubules. The GSH is not universal among prokaryotes. Entamoeba histolytica does not produce GSH or key enzymes of GSH metabolism. A general method of thiol analysis based upon fluorescent labeling with monobromobimane and HPLC separation of the resulting thiol derivatives was developed to determine the occurrence of GSH and other low molecular weight thiols in bacteria. Glutathione is the major thiol in cyanobacteria and in most bacteria closely related to the purple photosynthetic bacteria, but GSH was not found in archaebacteria, green bacteria, or GRAM positive bacteria. It suggested that glutathione metabolism was incorporated into eukaryotes at the time that mitochondria and chloroplasts were acquired by endosymbiosis. In Gram positive aerobes, coenzyme A occurs at millimolar levels and CoA disulfide reductases are identified. The CoA, rather than glutathione, may function in the oxygen detoxification processes of these organisms.

Fahey, R.↗

NASA NTRS1972

GLC of amino acids - A survey of contamination.

Analyses of biological substances and geochemical samples, of both terrestrial and extraterrestrial origin, are reported for amino acids at the 1-10 nanogram per gram level achieved by gas-liquid and ion-exchange chromatographic methods. These studies have shown that nanogram quantities of amino acids, present in water extracts of geochemical samples or in other samples of low amino acid concentration, can be successfully determined only if the researcher is well aware of the possible sources of contamination.

Rash, J. J.↗

NASA NTRS1989

Frequency response of electrochemical cells

Impedance concepts can be applied to the analysis of battery electrodes, yielding information about the structure of the electrode and the processes occurring in the electrode. Structural parameters such as the specific area (surface area per gram of electrode) can be estimated. Electrode variables such as surface overpotential, ohmic losses, and diffusion limitations may be studied. Nickel and cadmium electrodes were studied by measuring the ac impedance as a function of frequency, and the specific areas that were determined were well within the range of specific areas determined from BET measurements. Impedance spectra were measured for the nickel and cadmium electrodes, and for a 20 A-hr NiCd battery as functions of the state of charge. More work is needed to determine the feasibility of using frequency response as a nondestructive testing technique for batteries.

Thomas, Daniel L.↗

NASA NTRS1961

Effect of Inert, Reducing, and Oxidizing Atmospheres on Friction and Wear of Metals to 1000 F

Experiments were conducted in inert, reducing, and oxidizing atmospheres to determine their influence on the friction and wear properties of various metals. Nitrogen, argon, forming gas (10 volume percent H2, 90 volume percent N2), and various concentrations of oxygen in nitrogen were used. A 3/16-inch-radius hemispherical rider under a load of 1000 grams contacted the flat surface of a rotating disk. The surface speed employed was 35 feet per minute. The presence of surface oxides is vitally important to the protection of metals in sliding contact. Extremely high friction and excessive wear were encountered in the absence of these oxides. In some instances (electrolytically pure copper), the removal of the surface oxides resulted in mass welding of the specimens in sliding contact. Extremely small quantities of oxygen are sufficient to provide protection of metal surfaces; for example, with 440-C stainless steel, 0.03 volume percent oxygen was found to be adequate.

Buckley, Donald H.↗

DOE OSTI2024

Bactericidal effectors of the Stenotrophomonas maltophilia type IV secretion system: functional definition of the nuclease TfdA and structural determination of TfcB

ABSTRACT Stenotrophomonas maltophilia expresses a type IV protein secretion system (T4SS) that promotes contact-dependent killing of other bacteria and does so partly by secreting the effector TfcB. Here, we report the structure of TfcB, comprising an N-terminal domain similar to the catalytic domain of glycosyl hydrolase (GH-19) chitinases and a C-terminal domain for recognition and translocation by the T4SS. Utilizing a two-hybrid assay to measure effector interactions with the T4SS coupling protein VirD4, we documented the existence of five more T4SS substrates. One of these was protein 20845, an annotated nuclease. A S. maltophilia mutant lacking the gene for 20845 was impaired for killing Escherichia coli , Klebsiella pneumoniae , and Pseudomonas aeruginosa . Moreover, the cloned 20845 gene conferred robust toxicity, with the recombinant E. coli being rescued when 20845 was co-expressed with its cognate immunity protein. The 20845 effector was an 899 amino-acid protein, comprised of a GHH-nuclease domain in its N-terminus, a large central region of indeterminant function, and a C-terminus for secretion. Engineered variants of the 20845 gene that had mutations in the predicted catalytic site did not impede E. coli , indicating that the antibacterial effect of 20845 involves its nuclease activity. Using flow cytometry with DNA staining, we determined that 20845, but not its mutant variants, confers a loss in DNA content of target bacteria. Database searches revealed that uncharacterized homologs of 20845 occur within a range of bacteria. These data indicate that the S. maltophilia T4SS promotes interbacterial competition through the action of multiple toxic effectors, including a potent, novel DNase. IMPORTANCE Stenotrophomonas maltophilia is a multi-drug-resistant, Gram-negative bacterium that is an emerging pathogen of humans. Patients with cystic fibrosis are particularly susceptible to S. maltophilia infection. In hospital water systems and various types of infections, S. maltophilia co-exists with other bacteria, including other pathogens such as Pseudomonas aeruginosa . We previously demonstrated that S. maltophilia has a functional VirB/D4 type VI protein secretion system (T4SS) that promotes contact-dependent killing of other bacteria. Since most work on antibacterial systems involves the type VI secretion system, this observation remains noteworthy. Moreover, S. maltophilia currently stands alone as a model for a human pathogen expressing an antibacterial T4SS. Using biochemical, genetic, and cell biological approaches, we now report both the discovery of a novel antibacterial nuclease (TfdA) and the first structural determination of a bactericidal T4SS effector (TfcB).

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2022

Assessment of Bacterial Inoculant Delivery Methods for Cereal Crops

Despite growing evidence that plant growth-promoting bacteria can be used to improve crop vigor, a comparison of the different methods of delivery to determine which is optimal has not been published. An optimal inoculation method ensures that the inoculant colonizes the host plant so that its potential for plant growth-promotion is fully evaluated. The objective of this study was to compare the efficacy of three seed coating methods, seedling priming, and soil drench for delivering three bacterial inoculants to the sorghum rhizosphere and root endosphere. The methods were compared across multiple time points under axenic conditions and colonization efficiency was determined by quantitative polymerase chain reaction (qPCR). Two seed coating methods were also assessed in the field to test the reproducibility of the greenhouse results under non-sterile conditions. In the greenhouse seed coating methods were more successful in delivering the Gram-positive inoculant (Terrabacter sp.) while better colonization from the Gram-negative bacteria (Chitinophaga pinensis and Caulobacter rhizosphaerae) was observed with seedling priming and soil drench. This suggested that Gram-positive bacteria may be more suitable for the seed coating methods possibly because of their thick peptidoglycan cell wall. We also demonstrated that prolonged seed coating for 12 h could effectively enhance the colonization of C. pinensis, an endophytic bacterium, but not the rhizosphere colonizing C. rhizosphaerae. In the field only a small amount of inoculant was detected in the rhizosphere. This comparison demonstrates the importance of using the appropriate inoculation method for testing different types of bacteria for their plant growth-promotion potential.

Terrabacter↗

NASA NTRS2004

New method for estimating bacterial cell abundances in natural samples by use of sublimation

Evaluation Studies↗

NASA NTRSDate not supplied

The Effects of Chronic Sleep Restriction on Calorie and Macronutrient Intake

Introduction: Chronic sleep restriction (CSR) has been associated with increased calorie intake and increased consumption of fats and carbohydrates, with inconsistent changes in protein. However, the majority of studies have either been observational field studies with no sleep intervention, or laboratory-based studies where food availability may not have reflected participants’ real-world choices. We hypothesized that calorie, fat, and carbohydrate intake would increase during a week of imposed CSR compared to a week of sleep satiation (SS) among individuals living in their home environment. Methods: Twelve healthy participants (6 females) kept a fixed sleep-wake schedule, with a constant waketime, at home for four weeks (actigraphy confirmed compliance). During weeks one and three, participants maintained 9 hours in bed. During weeks two and four, participants were randomly assigned to experimental weeks of 5 and 9 hours of time-in-bed in a crossover design. Participants documented their food consumption during both experimental weeks using a picture-based meal logging application (MealLogger). Intake of calories and macronutrients were classified by two blinded evaluators. Descriptive statistics were calculated in SAS (Cary, NC). Results: Participants averaged 4.43 ± 0.33 (SD) hours of sleep per night during CSR compared to 7.42 ± 0.42 hours during SS. Participants consumed a daily average of 1812 ± 672 kilocalories, 71 ± 31 grams of total fat, 217 ± 69 grams of carbohydrates, and 84 ± 40 grams of protein during CSR, compared to 1682 ± 514 kilocalories, 68 ± 23 grams of total fat, 198 ± 61 grams of carbohydrates, and 77 ± 32 grams of protein during SS. Conclusion: Preliminary descriptive findings suggest that, on average, participants consumed more calories, from an increase in consumption of each macronutrient group, during a week of sleep restriction compared to a week of sleep satiation. Further analysis is needed to determine whether these differences are statistically different and to identify when calories were consumed in each of the experimental conditions.

chronic sleep restriction↗

NASA NTRS1988

Lewis Cliff 86010, a unique Antarctic meteorite: Possible new clues to the early history of the solar system

Results form the Antarctic sample, LEW 86010 indicate that it is an extremely interesting meteorite of clear igneous origin, and is probably closely related to ADOR. Several important questions await further detailed study. The most stringent test of the relationship to ADOR will come from the measurement of the oxygen isotopic composition. One possibility proposed is that LEW 86010 represents a partial or total melt of a mixture of material similar to the white Allende clasts and more ordinary chondritic meteorite material. Several lines of investigation will contribute to resolve this question. Further melting studies will be performed to determine whether LEW 86010 represents molten lava. In addition to test the connection with Allende, isotopic studies will look for anomalous isotopic compositions which are ubiquitous in Allende white inclusions. All of these studies will be performed on a tiny sample which weights only 5 grams, and is smaller than a marble, but which contains important clues to events which occurred during the birth of the solar system.

Mckay, G. A.↗

NASA NTRS1992

Mars global reference atmosphere model (Mars-GRAM)

Mars-GRAM is an empirical model that parameterizes the temperature, pressure, density, and wind structure of the Martian atmosphere from the surface through thermospheric altitudes. In the lower atmosphere of Mars, the model is built around parameterizations of height, latitudinal, longitudinal, and seasonal variations of temperature determined from a survey of published measurements from the Mariner and Viking programs. Pressure and density are inferred from the temperature by making use of the hydrostatic and perfect gas laws relationships. For the upper atmosphere, the thermospheric model of Stewart is used. A hydrostatic interpolation routine is used to insure a smooth transition from the lower portion of the model to the Stewart thermospheric model. Other aspects of the model are discussed.

Justus, C. G.↗

NASA NTRS2014

The Flux of Large Meteoroids Observed with Lunar Impact Monitoring

The flux of large meteoroids is not well determined due to relatively low number statistics, due mainly to the lack of collecting area available to meteor camera systems (10(2)-10(5) km2). Larger collecting areas are needed to provide reasonable statistics for flux calculations. The Moon, with millions of square kilometers of lunar surface, can be used as a detector for observing the population of large meteoroids in the tens of grams to kilogram mass range. This is accomplished by observing the flash of light produced when a meteoroid impacts the lunar surface, converting a portion of its kinetic energy to visible light detectable from Earth. A routine monitoring program at NASA's Marshall Space Flight Center has recorded over 300 impact flashes since early 2006. The program utilizes multiple 0.35 m (14 inch) Schmidt-Cassegrain telescopes, outfitted with video cameras using the 1/2 inch Sony EXview HAD CCDTM chip, to perform simultaneous observations of the earthshine hemisphere of the Moon when the lunar phase is between 0.1 and 0.5. This optical arrangement permits monitoring of approximately 3.8x10(6) km2 of lunar surface. A selection of 126 flashes recorded in 266.88 hours of photometric skies was analyzed, creating the largest and most homogeneous dataset of lunar impact flashes to date. Standard CCD photometric techniques outlined in [1] were applied to the video to determine the luminous energy, kinetic energy, and mass for each impactor, considering a range of luminous efficiencies. The flux to a limiting energy of 2.5x10(-6) kT TNT or 1.05×10(7) J is 1.03×10(-7) km(-2) hr(-1) and the flux to a limiting mass of 30 g is 6.14×10(-10) m(-2) yr(-1). Comparisons made with measurements and models of the meteoroid population indicate that the flux of objects in this size range is slightly lower (but within the error bars) than the power law distribution determined for the near Earth object population by [2].

Cooke, W. J.↗

DOE OSTI2025

A widespread family of molecular chaperones promotes the intracellular stability of type VIIb secretion system– exported toxins

To survive in highly competitive environments, bacteria use specialized secretion systems to deliver antibacterial toxins into neighboring cells, thereby inhibiting their growth. In many Gram-positive bacteria, the export of such toxins requires a membrane-bound molecular apparatus known as the type VIIb secretion system (T7SSb). Recently, it was shown that toxin recruitment to the T7SSb requires a physical interaction between a toxin and two or more so-called targeting factors, which harbor key residues required for T7SS-dependent protein export. However, in addition to these targeting factors, some toxins additionally require a protein belonging to the DUF4176 protein family. Here, by examining two toxin–DUF4176 protein pairs, we demonstrate that DUF4176 constitutes a family of toxin-specific molecular chaperones. In addition to being required for toxin stability in producing cells, we find that DUF4176 proteins facilitate toxin export by specifically interacting with a previously uncharacterized intrinsically disordered region found in many T7SS toxins. Using X-ray crystallography, we determine structures of several DUF4176 chaperones in their unbound state, and of a DUF4176 chaperone in complex with the binding site of its cognate toxin. These structures reveal that this binding site consists of a disordered amphipathic α-helix that requires interaction with its cognate chaperone for proper folding. Overall, we have identified a family of secretion system associated molecular chaperones found throughout T7SSb-containing Gram-positive bacteria.

Gkragkopoulou, Polyniki↗

NASA NTRS2004

A New Method for Estimating Bacterial Abundances in Natural Samples using Sublimation

We have developed a new method based on the sublimation of adenine from Escherichia coli to estimate bacterial cell counts in natural samples. To demonstrate this technique, several types of natural samples including beach sand, seawater, deep-sea sediment, and two soil samples from the Atacama Desert were heated to a temperature of 500 C for several seconds under reduced pressure. The sublimate was collected on a cold finger and the amount of adenine released from the samples then determined by high performance liquid chromatography (HPLC) with UV absorbance detection. Based on the total amount of adenine recovered from DNA and RNA in these samples, we estimated bacterial cell counts ranging from approx. l0(exp 5) to l0(exp 9) E. coli cell equivalents per gram. For most of these samples, the sublimation based cell counts were in agreement with total bacterial counts obtained by traditional DAPI staining. The simplicity and robustness of the sublimation technique compared to the DAPI staining method makes this approach particularly attractive for use by spacecraft instrumentation. NASA is currently planning to send a lander to Mars in 2009 in order to assess whether or not organic compounds, especially those that might be associated with life, are present in Martian surface samples. Based on our analyses of the Atacama Desert soil samples, several million bacterial cells per gam of Martian soil should be detectable using this sublimation technique.

Glavin, Daniel P.↗

DOE OSTI2025

Characterization of Tank 9H Annulus Sample in Support of Residual Material Inventory Determinations

The Savannah River National Laboratory (SRNL) was requested by Savannah River Mission Completion (SRMC) to provide sample preparation and characterization of the Tank 9H annulus sample in support of Residual Material Inventory Determinations. One Tank 9H sample in three vials [HTF-9-25-13, HTF-9-25-14 and HTF-9-25-15], with each vial containing approximately 200 mL of the Tank 9H annulus salt solution, were delivered to the SRNL Shielded Cells for sample preparation and characterizations in February 2025. The density of the “as-received” solution contained in each of the three Tank 9H annulus sample vials were determined followed by a solid-liquid separation on each one using 0.45-micron Nalgene® nylon filter membranes. The resulting filtrates were combined to form the Tank 9H annulus sample with a total volume of about 600 mL. The combined wet solid fractions, about a total of 4.8 grams of salt material, remaining on the filter membranes were air-dried in the Shielded Cells for 72 hours. The total weight of the air-dried solids was 2.1 grams. These air-dried solids were washed with deionized water (DI water) at a phase ratio of 60 mL DI water/gram of solids to recover insoluble solids, if any. No visible or measurable quantity of insoluble solids were recovered after DI water washing of the air-dried solids because the air-dried solids completely dissolved in the DI water. The solid fraction-wash water was not combined with the 600 mL of the filtrate solution, and the resulting solution was not screened or analyzed for radionuclides. Aliquot sample volumes of the undiluted Tank 9H annulus sample were sent to the SRNL analytical services groups for radionuclides, elementals, anions and total mercury analysis by various methods including radiochemical separations/counting methods, inductively coupled plasma-atomic emission spectroscopy (ICP-AES), and Inductively Coupled Plasma Mass Spectroscopy (ICP-MS) and special preparations. All sample analyses were performed in triplicate. This report presents the analytical characterization results for the Tank 9H annulus sample. The results are also reported where analytical methods yielded additional analytes, other than those requested by SRMC. In the characterization of the Tank 9H annulus sample, the detection limits for all the analytes, as specified in the Technical Task Request (TTR) and Task Technical and Quality Assurance Plan (TTQAP), were met.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

DOE OSTI2020

Scaling Trivalent Actinide and Lanthanide Recovery by Diglycolamide Resin from Savannah River Site’s Mark-18A Targets

The Savannah River National Laboratory is implementing a separation flowsheet to recover rare isotopes and transplutonium elements from irradiated 242 Pu targets known as Mark-18A targets. The Mark-18A targets contain the United States’ supply of nonseparated 244 Pu, which has a wide range of applications from nuclear forensics to production of superheavy elements such as flerovium. The targets also contain hundreds of grams of heavy curium ( 246–248 Cm), which is used as a target material for 252 Cf production. This work investigates the use of diglycolomide resin (DGA Resin) to recover valuable trivalent actinides from the Mark-18A targets. Batch contact experiments were performed on a representative simulant to determine mass loadings. Furthermore, the resin showed an overall capacity of 11 mg/mL for a mixed metal matrix (Zr and La–Gd). Column experiments showed chromatographic separation with transition-metal breakthroughs occurring first followed by the lanthanide series La–Gd. The experiments showed that lanthanide breakthrough occurred after 11 mg/mL mass loading was reached on the column with the mixed metal matrix. A radiological column experiment with an in-line UV/vis cell was able to detect Nd breakthrough just prior to 241 Am breakthrough. Implementing an in-line UV/vis cell into full-scale Mark-18A target processing will be used to limit breakthrough of trivalent actinides recovered from the targets.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2024

The Impact of Sorbent Amendments for Mercury Remediation on the Viability of Soil Microorganisms

Mercury (Hg) remediation across contaminated environments in the United States is an ongoing project. As part of the Hg cleanup strategy at East Fork Poplar Creek (EFPC), located in Oak Ridge, TN, the deployment of sorbents is considered. However, the impact of sorbents on soil microorganisms is poorly understood. In this study, we investigated the effect of sorbents on soil microorganism viability and biofilm formation to assess soil health during sorbent application for Hg remediation. We specifically investigated the effect of two engineered sorbents, Organoclay PM-199 and Organoclay MRM (which are manufactured from clay minerals formulated for various remediation applications), on two gram-negative organisms ( Serratia marcescens and Burkholderia thailandensis ) isolated from the Hg-contaminated EFPC bank soil. Pure cultures of S. marcescens or B. thailandensis were amended with 5% (w/v) and 25% (w/v) PM-199 and MRM, respectively, for 9 days. The samples were harvested, and bacterial cell viability was determined using a BacLight staining kit. Results showed that the growth of sorbent-amended S. marcescens was inhibited in contrast to that of unamended control. Furthermore, biochemical assays were used to analyze bacterial biofilm formation and integral biofilm components. Our results suggest that biofilm formation by sorbent-amended S. marcescens was negatively affected. In contrast, B. thailandensis amended with low concentrations of MRM showed enhanced growth and notable differences in biofilm morphology. These results suggest that the use of organoclay PM-199 and MRM at higher concentrations in field studies may hinder the growth of specific soil microorganisms.

54 ENVIRONMENTAL SCIENCES↗

NASA NTRS2019

How Does Water Delivery System Design Impact the Microbial Load of Salad Crops?

In a microgravity setting, such as the environment aboard the International Space Station (ISS), an ideal plant water delivery system is one that can grow edible crops with minimal resource consumption and minimal risk to crew members. There are also concerns associated with the ability to control fluid escape and biofilm formation resulting in potential dangers to systems, crops, or crewmembers. To identify an appropriate system, candidate systems were assembled and operated under simulated ISS environmental conditions (T,CO2,and RH) with red romaine lettuce (Lactuca sativa cultivar 'Outredgeous') as a model crop. Fluid reservoirs and randomly selected planting sites were sampled every seven days until maturity at which point edible plant biomass and root samples were also taken. Heterotrophic bacteria and fungi growth patterns throughout each planting cycle were determined by plate counts on appropriate agar media. The candidate systems were compared to a classic hydroponics system as a control and harvested crops were compared to controls as well as Veggie-grown and market produce. Plants harvested from candidate systems yielded lower average heterotrophic bacteria and fungi per gram of plant mass levels when compared to market and Veggie samples as well as those from the control system. Additional studies to evaluate the system sanitation regimen as well as testing additional crops should be considered to aid in the selection of an ideal system.

Curry, Aaron B.↗

DOE OSTI2022

‘ Candidatus Liberibacter asiaticus’-Encoded BCP Peroxiredoxin Suppresses Lipopolysaccharide-Mediated Defense Signaling and Nitrosative Stress In Planta

The lipopolysaccharides (LPS) of gram-negative bacteria trigger a nitrosative and oxidative burst in both animals and plants during pathogen invasion. Liberibacter crescens strain BT-1 is a surrogate for functional genomic studies of the uncultured pathogenic ‘Candidatus Liberibacter’ spp. that are associated with severe diseases such as citrus greening and potato zebra chip. Structural determination of L. crescens LPS revealed the presence of a very long chain fatty acid modification. L. crescens LPS pretreatment suppressed growth of Xanthomonas perforans on nonhost tobacco (Nicotiana benthamiana) and X. citri subsp. citri on host orange (Citrus sinensis), confirming bioactivity of L. crescens LPS in activation of systemic acquired resistance (SAR). L. crescens LPS elicited a rapid burst of nitric oxide (NO) in suspension cultured tobacco cells. Pharmacological inhibitor assays confirmed that arginine-utilizing NO synthase (NOS) activity was the primary source of NO generation elicited by L. crescens LPS. LPS treatment also resulted in biological markers of NO-mediated SAR activation, including an increase in the glutathione pool, callose deposition, and activation of the salicylic acid and azelaic acid (AzA) signaling networks. Transient expression of ‘Ca. L. asiaticus’ bacterioferritin comigratory protein (BCP) peroxiredoxin in tobacco compromised AzA signaling, a prerequisite for LPS-triggered SAR. Western blot analyses revealed that ‘Ca. L. asiaticus’ BCP peroxiredoxin prevented peroxynitrite-mediated tyrosine nitration in tobacco. ‘Ca. L. asiaticus’ BCP peroxiredoxin (i) attenuates NO-mediated SAR signaling and (ii) scavenges peroxynitrite radicals, which would facilitate repetitive cycles of ‘Ca. L. asiaticus’ acquisition and transmission by fecund psyllids throughout the limited flush period in citrus.

59 BASIC BIOLOGICAL SCIENCES↗