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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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At least 109 records · Page 6

A Bright, Photostable, and Far-Red Dye That Enables Multicolor, Time-Lapse, and Super-Resolution Imaging of Acidic Organelles

Lysosomes have long been known for their acidic lumens and efficient degradation of cellular byproducts. In recent years, it has become clear that their function is far more sophisticated, involving multiple cell signaling pathways and interactions with other organelles. Unfortunately, their acidic interior, fast dynamics, and small size make lysosomes difficult to image with fluorescence microscopy. Here we report a far-red small molecule, HMSiR 680 -Me, that fluoresces only under acidic conditions, causing selective labeling of acidic organelles in live cells. HMSiR 680 -Me can be used alongside other far-red dyes in multicolor imaging experiments and is superior to existing lysosome probes in terms of photostability and maintaining cell health and lysosome motility. We demonstrate that HMSiR 680 -Me is compatible with overnight time-lapse experiments as well as time-lapse super-resolution microscopy with a frame rate of 1.5 fps for at least 1000 frames. HMSiR 680 -Me can also be used alongside silicon rhodamine dyes in a multiplexed super-resolution microscopy experiment to visualize interactions between mitochondria and lysosomes with only a single excitation laser and simultaneous depletion. We envision this dye permitting a more detailed study of the role of lysosomes in dynamic cellular processes and disease.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Harnessing Heterologous Bacterial Two-Component Systems as Biosensors to Address Challenges in Fermentation Scale-Up

Scaling up bacterial fermentation from bench to industrial scale often results in unpredictable performance losses, possibly in part due to changes in microenvironmental conditions such as pH. To investigate this, we developed a suite of pH-sensitive biosensors from bacterial two-component systems (TCSs) that provide a dynamic, fluorescent readout in response to extracellular pH changes. TCSs consist of a periplasmic sensor histidine kinase (HK) that, in response to an extracellular stimulus, autophosphorylates intracellularly and subsequently transfers the phosphate to a cognate response regulator (RR) that modulates transcription of target genes. We utilized three pH-responsive TCSs (referred to here as CVJ1, CVJ30, and CVJ79) and linked their output to GFP. This was achieved by placing the RR promoter upstream of GFP or by constructing a chimeric RR composed of the native receiver domain and the DNA-binding domain of another well-characterized RR with a defined promoter. All components - HK, RR (native or chimeric), and GFP under its corresponding promoter - were cloned into a broad-host-range plasmid. Sensors were validated in Escherichia coli and Pseudomonas putida, including the muconic acid-producing strain P. putida TL207. All three biosensors successfully reported pH, with fluorescence (normalized to optical density) correlating strongly with media pH. Among the native sensors, CVJ79 showed the most robust performance while CVJ1 also performed best in its native form; CVJ30 exhibited improved functionality as a chimera, suggesting that modular RR design can enhance compatibility in some heterologous hosts. Further, CVJ79 was activated by alkaline conditions, while CVJ30 responded to acidic environments. Notably, CVJ1 was induced by high pH in wild-type E. coli and P. putida, but low pH in TL207. The observed differences in sensor activation between strains - particularly the divergent response of CVJ1 - suggest that host-specific regulatory pathways may influence how cells perceive and adapt to pH stress. Moving forward, these biosensors can be used to guide the rational design of more robust strains, optimize process conditions in real time, and inform strategies to minimize physiological heterogeneity during scale-up. Integrating these tools into high-throughput screening and bioreactors will be a key step toward improving predictability and performance in industrial bioprocesses.

09 BIOMASS FUELS↗

Optical Reflectance and Fluorescence for Detecting Nitrogen Needs in Zea mays L.

Nitrogen (N) status in field grown corn (Zea mays L.) was assessed using spectral techniques. Passive reflectance remote sensing and, both passive and active fluorescence sensing methods were investigated. Reflectance and fluorescence methods are reported to detect changes in the primary plant pigments (chlorophylls a and b; carotenoids) in higher plant species. As a general rule, foliar chlorophyll a (Chl a) and chlorophyll b (Chl b) usually exist in approx.3:l ratio. In plants under stress, Chl b content is affected before Chl a reductions occur. For reflectance, a version of the chlorophyll absorption in reflectance index (CARI) method was tested with narrow bands from the Airborne Imaging Spectroradiometer for Applications (ASIA). CARI minimizes the effects of soil background on the signal from green canopies. A modified CARI (MCARI) was used to track total Chl a levels in the red dip of the spectrum from the corn canopy. A second MCARI was used to track the auxiliary plant pigments (Chl b and the carotenoids) in the yellow/orange/red edge part of the reflectance spectrum. The difference between these two MCARI indices detected variations in N levels across the field plot canopies using ASIA data. At the leaf level, ratios of fluorescence emissions in the blue, green, red and far-red wavelengths sensed responses that were associated with the plant pigments, and were indicative of energy transfer in the photosynthetic process. N stressed corn stands could be distinguish from those with optimally applied N with fluorescence emission spectra obtained from individual corn leaves. Both reflectance and fluorescence methods are sensitive in detecting corn N needs and may be especially powerful in monitoring crop conditions if both types of information can be combined.

McMurtrey, J. E.↗

Click-Chemistry-Based Free Azide versus Azido Sugar Detection Enables Rapid In Vivo Screening of Glycosynthase Activity

Engineering of carbohydrate-active enzymes such as glycosynthases to enable chemoenzymatic synthesis of bespoke oligosaccharides has been limited by the lack of suitable ultrahigh-throughput screening methods capable of robustly detecting either starting substrates or end-products of the glycosidic bond formation reaction. Currently, there are limited screening methods available for rapid and highly sensitive single-cell-based screening of glycosynthase enzymes employing azido sugars as activated donor glycosyl substrates. Here, we report a fluorescence-based approach employing click-chemistry for the selective detection of glycosyl azides as substrates versus free inorganic azides as reaction products that facilitated an ultrahigh-throughput in vivo single-cell-based assay of glycosynthase activity. This assay was developed based on the distinct differences observed in relative fluorescence intensity of the triazole-containing fluorophore product formed during the click-chemistry reaction of organic glycosyl azides versus inorganic azides. This discovery formed the basis for proof of concept validation of a directed evolution methodology for screening and sorting glycosynthase mutants capable of synthesis of targeted fucosylated oligosaccharides. Our screening approach facilitated fluorescence-activated cell sorting of an error-prone polymerase chain reaction-based mutant library of fucosynthases expressed in Escherichia coli to identify several novel mutants that showed increased activity for ß-fucosyl azide-activated donor sugars toward desired acceptor sugars (e.g., pNP-xylose and lactose). Finally, we discuss avenues for improving this proof of concept in vivo assay method to identify better glycosynthase mutants and further demonstrate the broader applicability of this screening methodology for synthesis of bespoke glycans.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

An essential role for tungsten in the ecology and evolution of a previously uncultivated lineage of anaerobic, thermophilic Archaea

Trace metals have been an important ingredient for life throughout Earth’s history. Here, we describe the genome-guided cultivation of a member of the elusive archaeal lineage Caldarchaeales (syn. Aigarchaeota), Wolframiiraptor gerlachensis, and its growth dependence on tungsten. A metagenome-assembled genome (MAG) of W. gerlachensis encodes putative tungsten membrane transport systems, as well as pathways for anaerobic oxidation of sugars probably mediated by tungsten-dependent ferredoxin oxidoreductases that are expressed during growth. Catalyzed reporter deposition-fluorescence in-situ hybridization (CARD-FISH) and nanoscale secondary ion mass spectrometry (nanoSIMS) show that W. gerlachensis preferentially assimilates xylose. Phylogenetic analyses of 78 high-quality Wolframiiraptoraceae MAGs from terrestrial and marine hydrothermal systems suggest that tungsten-associated enzymes were present in the last common ancestor of extant Wolframiiraptoraceae. Our observations imply a crucial role for tungsten-dependent metabolism in the origin and evolution of this lineage, and hint at a relic metabolic dependence on this trace metal in early anaerobic thermophiles.

59 BASIC BIOLOGICAL SCIENCES↗

Negative thermal quenching of photoluminescence in a copper–organic framework emitter

Negative thermal quenching (NTQ), an abnormal phenomenon that the intensity of photoluminescence (PL) increases with increasing temperature, has essentially been restricted to either bulk semiconductors or very low temperatures. Here, we report a delayed fluorescence copper–organic framework exhibiting negative thermal quenching (NTQ) of photoluminescence, which is driven by the fluctuation between the localized and delocalized form of its imidazole ligand. The process is completely reversible on cooling/heating cycles. This study opens a new avenue to explore the electronically switchable NTQ effect in coordination networks and further to develop the NTQ-based light-emitting diodes.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Carbazole-substituted benzobisoxazoles: near-UV fluorescent emitters and ambipolar hosts for organic light-emitting diodes

The design of organic light emitting diode (OLED) materials with the potential for exhibiting thermally-activated delayed fluorescence (TADF) is reported. Using computational methods (DFT/TD-DFT) as a guiding tool, six materials with a benzobisoxazole (BBO) core and donor–acceptor–donor architectures were designed by changing the conjugation position of carbazole-substituted phenyl substituents and the type of BBO isomer (cis- vs. trans-). Experimental steady-state and transient absorption spectroscopic techniques were utilized to probe the TADF activity of these molecules. Each material was then used in host–guest OLED devices as either near-UV dopants or host with low singlet-triplet energy differences. The electroluminescent properties show that when used as dopants these materials provide near-UV emission (CIE y < 0.06 and CIE x = 0.16), whereas when used as hosts, these materials show reduced operating voltages and increased performance efficiencies when compared to commercial materials.

36 MATERIALS SCIENCE↗

Peeling back the many layers of competitive exclusion

Baby chicks administered a fecal transplant from adult chickens are resistant to Salmonella colonization by competitive exclusion. A two-pronged approach was used to investigate the mechanism of this process. First, Salmonella response to an exclusive (Salmonella competitive exclusion product, Aviguard ® ) or permissive microbial community (chicken cecal contents from colonized birds containing 7.85 Log 10 Salmonella genomes/gram) was assessed ex vivo using a S. typhimurium reporter strain with fluorescent YFP and CFP gene fusions to rrn and hilA operon, respectively. Second, cecal transcriptome analysis was used to assess the cecal communities’ response to Salmonella in chickens with low (≤5.85 Log 10 genomes/g) or high (≥6.00 Log 10 genomes/g) Salmonella colonization. The ex vivo experiment revealed a reduction in Salmonella growth and hilA expression following co-culture with the exclusive community. The exclusive community also repressed Salmonella’s SPI-1 virulence genes and LPS modification, while the anti-virulence/inflammatory gene avrA was upregulated. Salmonella transcriptome analysis revealed significant metabolic disparities in Salmonella grown with the two different communities. Propanediol utilization and vitamin B12 synthesis were central to Salmonella metabolism co-cultured with either community, and mutations in propanediol and vitamin B12 metabolism altered Salmonella growth in the exclusive community. There were significant differences in the cecal community’s stress response to Salmonella colonization. Cecal community transcripts indicated that antimicrobials were central to the type of stress response detected in the low Salmonella abundance community, suggesting antagonism involved in Salmonella exclusion. This study indicates complex community interactions that modulate Salmonella metabolism and pathogenic behavior and reduce growth through antagonism may be key to exclusion.

59 BASIC BIOLOGICAL SCIENCES↗

Aerosol Microphysics and Chemical Measurements at Mt. Soledad and Scripps Pier during the Eastern Pacific Cloud Aerosol Precipitation Experiment (EPCAPE) from February 2023 to February 2024 UCSD Library Collection

This dataset includes guest instrument measurements and other PI products for aerosol microphysics and chemical measurements collected at Mt. Soledad and Scripps Pier during the Eastern Pacific Cloud Aerosol Precipitation Experiment (EPCAPE) from February 2023 to February 2024. The measurements include the following instruments at Mt. Soledad: High-Resolution Time-of-Flight Aerosol Mass Spectrometer (HR-ToF-AMS, Aerodyne), Scanning Electrical Mobility Spectrometer (SEMS, Brechtel Manufacturing Inc.), Aerodynamic Particle Sizer (APS, Droplet Measurements Technologies), Single Particle Soot Photometer (SP2, Drople Measurements Technologies), Meteorological Station (WXT520, Vaisala), Ozone (Teco), and trace gas proxies (Teledyne). In addition, the analyses of particle filters collected at Mt. Soledad for three dry-diameter size cuts (<1 micron, <0.5 micron, <0.18 micron) and at Scripps Pier for one dry-diameter size cut (<1 micron) by Fourier Transform Infrared (FTIR) and X-ray Fluorescence (XRF) are reported. A differential mobility analyzer operated as a scanning mobility particle sizer (SMPS, TSI Inc.), a printed particle optical spectrometer (POPS, Grimm), and a continuous flow diffusion cloud condensation nuclei (CCN, DMT) counter provide the mobility aerosol size distribution (30-360 nm), optical size distribution (150 - 6000 nm), size-resolved CCN distribution (30-360 nm) at 0.2, 0.4, 0.6, 0.8, and 1.0% supersaturation. Measurements are reported for both sampling from an isokinetic aerosol inlet and from a Counterflow Virtual Impactor (CVI, Brechtel Manufacturing Inc.). The data are available at the following link: https://library.ucsd.edu/dc/collection/bb0898306q

54 ENVIRONMENTAL SCIENCES↗

Absolute concentrations of O/3P/ in the lower thermosphere at night

A rocketborne instrument module for detecting atomic oxygen concentrations by the resonance fluorescence technique is reported. The module contained a source lamp emitting O I 1304 A photons and a detector for the scattered photons. The module was constructed so that it could be coupled to a fast flow reactor for calibration purposes. During the flight the module was radially deployed from the payload on an oscillating boom with a maximum extension of 90 cm from the skin. Analysis of the initial flight data indicates that the shock formed by the motion of the rocket had a substantial effect on the data and the maximum concentration of the layer at 93 km was about 1.1 x 10 to the 11th/cu cm.

Sharp, W. E.↗

Quantitative measurement of transverse injector and free stream interaction in a nonreacting SCRAMJET combustor using laser-induced iodine fluorescence

A preliminary quantitative study of the compressible flowfield in a steady, nonreacting model SCRAMJET combustor using laser-induced iodine fluorescence (LIIF) is reported. Measurements of density, temperature, and velocity were conducted with the calibrated, nonintrusive, optical technique for two different combustor operating conditions. First, measurements were made in the supersonic flow over a rearward-facing step without transverse injection for comparison with calculated pressure profiles. The second configuration was staged injection behind the rearward-facing step at an injection dynamic pressure ratio of 1.06. These experimental results will be used to validate computational fluid dynamic (CFD) codes being developed to model supersonic combustor flowfields.

Fletcher, D. G.↗

The emergence of X-rays and gamma-rays from supernova 1987A

Attempts to arrive at a unified scenario for the optical, X-ray, and gamma-ray emission of SN 1987A are discussed. Theoretical spectra are in reasonable agreement with recent observations of hard X-rays by the Ginga and Mir satellites, but the early turn-on of X-rays suggests that the mantle and envelope may be 'leaky', perhaps as a result of Rayleigh-Taylor instabilities and clumping. The soft X-ray spectrum should be dominated by a 6.4 keV Fe K-alpha fluorescence line. The reported Ginga detection of 4-10 keV X-ray emission is also discussed.

Shull, J. Michael↗

Membrane association of sucrose synthase: changes during the graviresponse and possible control by protein phosphorylation

Sucrose synthase (SuSy) plays an important role in sucrose degradation and occurs both as a soluble and as a membrane-associated enzyme in higher plants. We show that membrane association can vary in vivo in response to gravistimulation, apparently involving SuSy dephosphorylation, and is a reversible process in vitro. Phosphorylation of SuSy has little effect on its activity but decreases its surface hydrophobicity as reported with the fluorescent probe bis-ANS. We postulate that phosphorylation of SuSy (and perhaps other membrane proteins) is involved in the release of the membrane-bound enzyme in part as a result of decreased surface hydrophobicity.

Non-NASA Center↗

Time-Resolved Ultraviolet Spectroscopy of The M-Dwarf GJ 876 Exoplanetary System

Extrasolar planets orbiting M-stars may represent our best chance to discover habitable worlds in the coming decade. The ultraviolet spectrum incident upon both Earth-like and Jovian planets is critically important for proper modeling of their atmospheric heating and chemistry. In order to provide more realistic inputs for atmospheric models of planets orbiting low-mass stars, we present new near- and far-ultraviolet (NUV and FUV) spectroscopy of the M-dwarf exoplanet host GJ 876 (M4V). Using the COS and STIS spectrographs on board the Hubble Space Telescope, we have measured the 1150-3140 A spectrum of GJ 876. We have reconstructed the stellar H1 Ly alpha emission line profile, and find that the integrated Ly alpha flux is roughly equal to the rest of the integrated flux (1150-1210 A + 1220-3140 A) in the entire ultraviolet bandpass (F(Ly alpha)/F(FUV+NUV) equals approximately 0.7). This ratio is approximately 2500x greater than the solar value. We describe the ultraviolet line spectrum and report surprisingly strong fluorescent emission from hot H2 (T(H2) greater than 2000 K). We show the light curve of a chromospheric + transition region flare observed in several far-UV emission lines, with flare/quiescent flux ratios greater than or equal to 10. The strong FUV radiation field of an M-star (and specifically Ly alpha) is important for determining the abundance of O2--and the formation of biomarkers-in the lower atmospheres of Earth-like planets in the habitable zones of low-mass stars.

France, Kevin↗

Time-Resolved Ultraviolet Spectroscopy of the M-Dwarf GJ 876 Exoplanetary System

Extrasolar planets orbiting M-stars may represent our best chance to discover habitable worlds in the coming decade. The ultraviolet spectrum incident upon both Earth-like and Jovian planets is critically important for proper modeling of their atmospheric heating and chemistry. In order to provide more realistic inputs for atmospheric models of planets orbiting low-mass stars, we present new near- and far-ultraviolet (NUV and FUV) spectroscopy of the M-dwarf exoplanet host GJ 876 (U4V). Using the COS and STIS spectrographs aboard the Hubble Space Telescope, we have measured the 1150 - 3140 Ang. spectrum of GJ 876. We have reconstructed the stellar H I Ly-alpha emission line profile, and find that the integrated Ly-apha flux is roughly equal to the rest of the integrated flux (1150 - 1210 Ang + 1220 - 3140 Ang) in the entire ultraviolet bandpass (F(Ly-alpha)/F(FUV+NUV) approximately equals 0.7). This ratio is approximately 2500 x greater than the solar value. We describe the ultraviolet line spectrum and report surprisingly strong fluorescent emission from hot H2 (T(H2) > 2000 K). We show the light-curve of a chromospheric + transition region flare observed in several far-UV emission lines, with flare/ quiescent flux ratios :2: 10. The strong FUV radiation field of an M-star (and specifically Ly-alpha) is important for determining the abundance of O2 - and the formation of biomarkers - in the lower atmospheres of Earth-like planets in the habitable zones of low-mass stars.

France, Kevin↗

Pioneering a Biobased UAS

With the exponential growth of interest in unmanned aerial vehicles (UAVs) and their vast array of applications in both space exploration and terrestrial uses such as the delivery of medicine and monitoring the environment, the 2014 Stanford-Brown-Spelman iGEM team is pioneering the development of a fully biological UAV for scientific and humanitarian missions. The prospect of a biologically-produced UAV presents numerous advantages over the current manufacturing paradigm. First, a foundational architecture built by cells allows for construction or repair in locations where it would be difficult to bring traditional tools of production. Second, a major limitation of current research with UAVs is the size and high power consumption of analytical instruments, which require bulky electrical components and large fuselages to support their weight. By moving these functions into cells with biosensing capabilities - for example, a series of cells engineered to report GFP, green fluorescent protein, when conditions exceed a certain threshold concentration of a compound of interest, enabling their detection post-flight - these problems of scale can be avoided. To this end, we are working to engineer cells to synthesize cellulose acetate as a novel bioplastic, characterize biological methods of waterproofing the material, and program this material's systemic biodegradation. In addition, we aim to use an "amberless" system to prevent horizontal gene transfer from live cells on the material to microorganisms in the flight environment. So far, we have: successfully transformed Gluconacetobacter hansenii, a cellulose-producing bacterium, with a series of promoters to test transformation efficiency before adding the acetylation genes; isolated protein bands present in the wasp nest material; transformed the cellulose-degrading genes into Escherichia coli; and we have confirmed that the amberless construct prevents protein expression in wild-type cells. In addition, as part of our human outreach project, we have been in touch with leaders in the fields of UAVs, synthetic biology, and earth sciences, and it is clear that biodegradable UAVs could have a significant impact on the industry.

Escherichia↗

Towards a Biosynthetic UAV

We are currently working on a series of projects towards the construction of a fully biological unmanned aerial vehicle (UAV) for use in scientific and humanitarian missions. The prospect of a biologically-produced UAV presents numerous advantages over the current manufacturing paradigm. First, a foundational architecture built by cells allows for construction or repair in locations where it would be difficult to bring traditional tools of production. Second, a major limitation of current research with UAVs is the size and high power consumption of analytical instruments, which require bulky electrical components and large fuselages to support their weight. By moving these functions into cells with biosensing capabilities - for example, a series of cells engineered to report GFP, green fluorescent protein, when conditions exceed a certain threshold concentration of a compound of interest, enabling their detection post-flight - these problems of scale can be avoided. To this end, we are working to engineer cells to synthesize cellulose acetate as a novel bioplastic, characterize biological methods of waterproofing the material, and program this material's systemic biodegradation. In addition, we aim to use an "amberless" system to prevent horizontal gene transfer from live cells on the material to microorganisms in the flight environment.

Biological↗

Multi Spectural Flourescence Imager (MSFI)

Genetic transformation with in vivo reporter genes for fluorescent proteins can be performed on a variety of organisms to address fundamental biological questions. Model organisms that may utilize an ISS imager include unicellular organisms (Saccharomyces cerevisiae), plants (Arabidopsis thaliana), and invertebrates (Caenorhabditis elegans).

Multi-Spectural↗