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DOE OSTI2022

Regulators of early maize leaf development inferred from transcriptomes of laser capture microdissection (LCM)-isolated embryonic leaf cells

The superior photosynthetic efficiency of C 4 leaves over C 3 leaves is owing to their unique Kranz anatomy, in which the vein is surrounded by one layer of bundle sheath (BS) cells and one layer of mesophyll (M) cells. Kranz anatomy development starts from three contiguous ground meristem (GM) cells, but its regulators and underlying molecular mechanism are largely unknown. To identify the regulators, we obtained the transcriptomes of 11 maize embryonic leaf cell types from five stages of pre-Kranz cells starting from median GM cells and six stages of pre-M cells starting from undifferentiated cells. Principal component and clustering analyses of transcriptomic data revealed rapid pre-Kranz cell differentiation in the first two stages but slow differentiation in the last three stages, suggesting early Kranz cell fate determination. In contrast, pre-M cells exhibit a more prolonged transcriptional differentiation process. Differential gene expression and coexpression analyses identified gene coexpression modules, one of which included 3 auxin transporter and 18 transcription factor (TF) genes, including known regulators of Kranz anatomy and/or vascular development. In situ hybridization of 11 TF genes validated their expression in early Kranz development. We determined the binding motifs of 15 TFs, predicted TF target gene relationships among the 18 TF and 3 auxin transporter genes, and validated 67 predictions by electrophoresis mobility shift assay. From these data, we constructed a gene regulatory network for Kranz development. Our study sheds light on the regulation of early maize leaf development and provides candidate leaf development regulators for future study.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2020

Direct evidence that tumor cells soften when navigating confined spaces.

The mechanical properties of cells strongly regulate many physiological and pathological processes. For example, in cancer, invasive and metastatic tumor cells have often been reported to be softer than nontumor cells, raising speculation that cancer cells might adaptively soften to facilitate migration through narrow tissue spaces. Despite growing interest in targeting cell softening to impede invasion and metastasis, it remains to be directly demonstrated that tumor cells soften as they migrate through confined spaces. Here, we address this open question by combining topographically patterned substrates with atomic force microscopy (AFM). Using a polydimethylsiloxane open-roof microdevice featuring tapered, fibronectin-coated channels, we followed the migration of U2OS cells through various stages of confinement while simultaneously performing AFM indentation. As cells progress from unconfined migration to fully confined migration, cells soften and exclude Yes-associated protein from the nucleus. Superresolution imaging reveals that confinement induces remodeling of actomyosin stress fiber architecture. Companion studies with flat one-dimensional microlines indicate that the changes in cytoarchitecture and mechanics are intrinsically driven by topographical confinement rather than changes in cellular aspect ratio. Our studies represent among the most direct evidence to date that tumor cells soften during confined migration and support cell softening as a mechanoadaptive mechanism during invasion.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2023

An invertible seven-dimensional Dirichlet cell characterization of lattices

Characterization of crystallographic lattices is an important tool in structure solution, crystallographic database searches and clustering of diffraction images in serial crystallography. Characterization of lattices by Niggli-reduced cells (based on the three shortest non-coplanar lattice vectors) or by Delaunay-reduced cells (based on four non-coplanar vectors summing to zero and all meeting at obtuse or right angles) is commonly performed. The Niggli cell derives from Minkowski reduction. The Delaunay cell derives from Selling reduction. All are related to the Wigner–Seitz (or Dirichlet, or Voronoi) cell of the lattice, which consists of the points at least as close to a chosen lattice point as they are to any other lattice point. The three non-coplanar lattice vectors chosen are here called the Niggli-reduced cell edges. Starting from a Niggli-reduced cell, the Dirichlet cell is characterized by the planes determined by 13 lattice half-edges: the midpoints of the three Niggli cell edges, the six Niggli cell face-diagonals and the four body-diagonals, but seven of the lengths are sufficient: three edge lengths, the three shorter of each pair of face-diagonal lengths, and the shortest body-diagonal length. These seven are sufficient to recover the Niggli-reduced cell.

Chemistry↗

DOE OSTI2020

Antibiotic tigecycline inhibits cell proliferation, migration and invasion via down‐regulating CCNE2 in pancreatic ductal adenocarcinoma

Abstract Recently, many researches have reported that antibiotic tigecycline has significant effect on cancer treatment. However, biomedical functions and molecular mechanisms of tigecycline in human pancreatic ductal adenocarcinoma (PDAC) remain unclear. In the current study, we tried to assess the effect of tigecycline in PDAC cells. AsPC‐1 and HPAC cells were treated with indicated concentrations of tigecycline for indicated time, and then, MTT, BrdU and soft agar assay were used to test cell proliferation. The effect of tigecycline on cell cycle and cellular apoptosis was tested by cytometry. Migration and invasion were detected by wound healing assay and transwell migration/invasion assay. Expressions of cell cycle‐related and migration/invasion‐related protein were determined by using Western blot. The results revealed that tigecycline observably suppressed cell proliferation by inducing cell cycle arrest at G0/G1 phase and blocked cell migration/invasion via holding back the epithelial‐mesenchymal transition (EMT) process in PDAC. In addition, tigecycline also remarkably blocked tumorigenecity in vivo. Furthermore, the effects of tigecycline alone or combined with gemcitabine in vitro or on PDAC xenografts were also performed. The results showed that tigecycline enhanced the chemosensitivity of PDAC cells to gemcitabine. Interestingly, we found CCNE2 expression was declined distinctly after tigecycline treatment. Then, CCNE2 was overexpressed to rescue tigecycline‐induced effect. The results showed that CCNE2 overexpression significantly rescued tigecycline‐inhibited cell proliferation and migration/invasion. Collectively, we showed that tigecycline inhibits cell proliferation, migration and invasion via down‐regulating CCNE2, and tigecycline might be used as a potential drug for PDAC treatment alone or combined with gemcitabine.

Yang, Jie↗

DOE OSTI2024

Data for Hydrothermal Conditioning of Oleaginous Yeast Cells to Enable Recovery of Lipids as Potential Drop-in Fuel Precursors

Lipids produced using oleaginous yeast cells are an emerging feedstock to manufacture commercially valuable oleochemicals ranging from pharmaceuticals to lipid-derived biofuels. Production of biofuels using oleaginous yeast is a multistep procedure that requires yeast cultivation and harvesting, lipid recovery, and conversion of the lipids to biofuels. The quantitative recovery of the total intracellular lipid from the yeast cells is a critical step during the development of a bioprocess. Their rigid cell walls often make them resistant to lysis. The existing methods include mechanical, chemical, biological and thermochemical lysis of yeast cell walls followed by solvent extraction. In this study, an aqueous thermal pretreatment was explored as a method for lysing the cell wall of the oleaginous yeast Rhodotorula toruloides for lipid recovery. Hydrothermal pretreatment for 60 min at 121 °C with a dry cell weight of 7% (w/v) in the yeast slurry led to a recovery of 84.6 ± 3.2% (w/w) of the total lipids when extracted with organic solvents. The conventional sonication and acid-assisted thermal cell lysis led to a lipid recovery yield of 99.8 ± 0.03% (w/w) and 109.5 ± 1.9% (w/w), respectively. The fatty acid profiles of the hydrothermally pretreated cells and freeze-dried control were similar, suggesting that the thermal lysis of the cells did not degrade the lipids. This work demonstrates that hydrothermal pretreatment of yeast cell slurry at 121 °C for 60 min is a robust and sustainable method for cell conditioning to extract intracellular microbial lipids for biofuel production and provides a baseline for further scale-up and process integration.

Conversion↗

DOE OSTI2025

Insights into regulatory T-cell and type-I interferon roles in determining abacavir-induced hypersensitivity or immune tolerance

Introduction Clinical use of several small molecule drugs may lead to severe T-cell-mediated idiosyncratic drug hypersensitivity reactions (iDHR) linked to HLA alleles, including abacavir (ABC) with HLA-B*57:01. Due to study limitations in humans, pathogenic networks in iDHR remain elusive. HLA transgenic murine models have been proposed to bridge knowledge gaps in tolerance and susceptibility to drugs. Methods Mice expressing HLA-B*57:01 and Foxp3-DTR/EGFP were generated to selectively deplete regulatory T-cells (Treg) with diphtheria toxin. ABC was administered for 8 days alone or together with cell- and cytokine-depleting antibodies. Cellular and transcriptomic responses were analyzed by RNA, flow cytometry and fluorescence methods. Results While CD8 + T-cell responses to ABC require HLA presentation, ABC also triggered mitochondrial stress in macrophagesin vitro, independently of HLA.In vivo, Treg were the primary mechanism of drug tolerance controlling HLA presentation and costimulation by antigen presenting cells. Treg ablation uncovered immune adverse events linked to activation and proliferation of both drug-specific and bystander CD8 + T-cells through CD28-mediated pathways with support from CD4 + non-Treg. Type-I interferon (IFN-I) and cellular-stress pathways influenced the fate of lymph node cells responding to ABC, implicating innate immune cells such as macrophages and plasmacytoid dendritic cells in the development of T-cell responses against the drug. IFN-I and IL-2 were necessary for CD8 + T-cell differentiation and ABC-induced adverse reactions. Conclusions This study unveils novel immune mechanisms driven by drug and host-related factors required forin vivoreactions and sheds light on potential biomarker and therapeutic targets for managing and preventing severe and life-threatening iDHR.

Immunology↗

NASA NTRS1963

The solar cells and their mounting

Objectives in development of the solar plant for the Telstar spacecraft were to provide a power source which would withstand launching stresses and the expected space environment, with optimum end-of-life performance. Radiation damage to the silicon solar cells is the primary factor limiting their useful life; the effect of energetic protons or electrons is the generation of recombination centers in the silicon which reduce the minority-carrier diffusion length and therefore the long-wave response of the cell. The spacecraft solar cells use the n-on-p structure, in preference to conventional p-on-n structure, to obtain a factor of 3 to 10 increased life expectancy. Response to light in the 0.4 to 0.7 micron range is enhanced by using a thin n-layer (about 0.5 micron) and an antireflection coating with minimum reflectance at 0.55 micron wavelength. Early estimates of electron and proton fluxes in the satellite orbit showed that even the best cells would not give sufficient life without radiation shielding. Therefore the cells are protected against electrons of energy up to 1 Mev by 0.3 gm/sq. cm sapphire cover plates. The cell mountings are designed to withstand peak vibration stresses of 200 g and repeated temperature cycles from +65°C to −100°C. The 3600-cell solar power plant is composed of 300 twelve-cell groups of 1 cm × 2 cm cells, yielding a nominal initial power of 14 watts at 28 volts for any spin-axis orientation relative to the sun. Telemetry information on performance of the solar plant indicates degradation of the shielded solar cells equal to that measured in the laboratory on unshielded cells with a 1-Mev normal incidence flux of 6 × 10^(12) electrons/(sq. cm day). From this comparison it is estimated that the plant will degrade to 68 per cent of its initial output after two years in orbit.

SPACECRAFT POWER SUPPLY↗

NASA NTRS1972

ATS-5 solar cell experiment results after one year in synchronous orbit

The results of the ATS-5 solar cell experiment after one year in synchronous orbit are reported. A partial failure in the experimental electronics package has caused a loss of data from half the 80 experimental solar cells. Procedures for extracting data due to a partial spacecraft failure are described and discussed. Data from the remaining 40 solar cells, including 15 mounted on a thin flexible structure are analyzed. Data are corrected to a solar intensity of 140 mW/sq cm and a temperature of 25 C. It was found that after one year in synchronous orbit: (1) cells with 1.52-mm-thick coverslides did not show a clear-cut advantage over those with 0.15-mm coverslides, (2) cells with solderless grid lines are degrading at the same rate as are cells with solder-dipped grid lines, (3) cells not quite completely covered with coverslides suffered a large power loss in comparison to cells fully covered, (4) no clear-cut advantage of 10-cm cells over 2-cm cells has yet been observed, (5) cells mounted on the flexible panel with relatively little backshielding did not degrade any faster than those with substantial backshielding, and (6) the flight data in large part confirms the adequacy of the ground-based techniques used in our preflight radiation test program.

Anspaugh, B. E.↗

NASA NTRS1971

Engineering experimental program on the effects of near-space radiation on lithium doped solar cells

The results of an experimental evaluation of the real-time degradation characteristics of lithium-diffused silicon solar cells are reported. A strontium-90 radioisotope was used for simulation of a typical earth-orbital electron environment. The experiment was performed in an ion pump vacuum chamber with samples maintained at -50, +20, +50, and +80 C. Samples were illuminated during the 6-month exposure run with solar cell 1-5 characteristics measured periodically in situ. This 6-month exposure corresponded to a 1 MeV equivalent fluence of approximately 10 to the 14th power electrons/sq cm. Several types of lithium cells were irradiatied and compared directly with conventional N/P cells. The best lithium cells compared favorably with N/P cells, particularly at the higher test temperatures. With a slight improvement of initial performance characteristics, lithium cells appear feasible for 5 to 10 year missions at synchronous altitude. Based on the reported results and those of other irradiation experiments, lithium cells would appear to be superior to N/P cells in proton-dominated earth-orbital environments. Another important conclusion of the effort was that illuminated/loaded cells degrade more rapidly than do dark/unloaded cells. The irradiation experiment provided data of high quality with a high degree of confidence because of the experimental and statistical analysis techniques utilized.

Source record↗

NASA NTRS1973

A simple method for the determination of effective rain cell dimensions and orientation

A simple method is proposed for the determination of effective rain cell dimensions and orientation. Two rain cell models were considered: the circular cell and the elliptical cell. In both models it is assumed that the rain rate is constant throughout the cell, that all cell locations are equally likely, and that the cell dimensions depend upon the rain rate. It is then shown that the effective cell dimensions and orientation may be deduced from rain rate statistics accumulated at two or three closely spaced sites, depending upon the model chosen. The results of this study alone with previous estimates of rain cell dimension indicate that the rain rate measurement points should be spaced on the order of one-half kilometer apart. The proposed method is simple and relatively inexpensive; thus, this approach is readily suited to the study of the dependence of rain cell size and orientation upon climatic region. The resulting rain cell characteristics are of direct value in the prediction of millimeter wavelength attenuation statistics on both single-terminal and diversity earth-space propagation paths as well as point-to-point terrestrial links.

Hodge, D. B.↗

NASA NTRS1976

A theoretical study of heterojunction and graded band gap type solar cells

The theory of heterojunction and graded bandgap solar cells was studied to help determine the material compositions and device dimensions needed for high efficiency solar cells. Because of the involved analysis of the fundamental equations describing solar cell operation, a general numerical device analysis program was used. A major part of the initial work was involved in modifying an existing silicon solar cell analysis program to account for the unique features of graded bandgap and heterojunction solar cells. The most successful III-V solar cells have so far been constructed in the GaAs and Gal-xAlxAs material systems; this project was concerned with such solar cells. The most efficient solar cell so far evaluated is an abrupt heterojunction cell with a pure AlAs layer at the surface with a GaAs substrate. The predicted efficiency for this cell is slightly larger than that of a graded bandgap Gal-xAlxAs solar cell.

Hauser, J. R.↗

NASA NTRS1980

Lightweight fuel cell powerplant components program

A lightweight hydrogen-oxygen alkaline fuel cell incorporated into the design of a lightweight fuel cell powerplant (LFCP) was analytically and experimentally developed. The powerplant operates with passive water removal which contributes to a lower system weight and extended operating life. A preliminary LFCP specification and design table were developed along with a lightweight power section for the LFCP design, consisting of repeating two-cell modules was designed. Two, four-cell modules were designed incorporating 0.508 sq ft active area space shuttle technology fuel cells. Over 1,200 hours of single-cell and over 8,800 hours of two-cell module testing was completed. The 0.25 sq ft active area lightweight cell design was shown to be capable of operating on propellant purity reactants out to a current density of 600ASF. Endurance testing of the two-cell module configuration exceeded the 2,500-hour LFCP voltage requirements out to 3700-hours. A two-cell module capable of operating at increased reactant pressure completed 1000 hours of operation at a 30 psia reactant pressure. A lightweight power section consisting of fifteen, two-cell modules connected electrically in series was fabricated.

Martin, R. E.↗

NASA NTRS1981

Coplanar back contacts for thin silicon solar cells

The type of coplanar back contact solar cell described was constructed with interdigitated n(+) and p(+) type regions on the back of the cell, such that both contacts are made on the back with no metallization grid on the front. This cell construction has several potential advantages over conventional cells for space use namely, convenience of interconnects, lower operating temperatures and higher efficiency due to the elimination of grid shadowing. However, the processing is more complex, and the cell is inherently more radiation sensitive. The latter problem can be reduced substantially by making the cells very thin (approximately 50 micrometers). Two types of interdigitated back contact cells are possible, the types being dependent on the character of the front surface. The front surface field cell has a front surface region that is of the same conductivity type as the bulk but is more heavily doped. This creates an electric field at the surface which repels the minority carriers. The tandem junction cell has a front surface region of a conductivity type that is opposite to that of the bulk. The junction thus created floats to open circuit voltage on illumination and injects carriers into the bulk which then can be collected at the rear junction. For space use, the front surface field cell is potentially more radiation resistant than the tandem junction cell because the flow of minority carriers (electrons) into the bulk will be less sensitive to the production of recombination centers, particularly in the space charge region at the front surface.

Storti, G.↗

NASA NTRS1982

Nondestructive evaluation techniques for nickel-cadmium aerospace battery cells

The ac impedance characteristics of Ni-Cd cells as an in-situ, nondestructive means of determining cell lifetime, particularly with respect to the probability of premature failure were evaluated. Emphasis was on evaluating Ni-Cd cell impedance over a wide frequency range (10,000 to 0.0004 Hz) as the cells were subjected to charge/discharge cycle testing. The results indicate that cell degradation is reflected in the low frequency (Warburg) impedance characteristics associated with diffusion processes. The Warburg slope (W) was found to steadily increase as a function of cell aging for completely discharged cells. In addition, based on data for two cells, a high or rapidly increasing value for W signals imminent cell failure by one mechanism. Degradation by another mechanism is apparently reflected in a fall-off (roll-over) of W at lower frequencies. As a secondary result, the frequency dependence of the absolute cell impedance at low frequencies (5 - 500 mHz) was found to be a good indication of the cell state-of-charge.

Haak, R.↗

NASA NTRS1987

Development of ambient temperature secondary lithium cells

JPL is developing ambient temperature secondary lithium cells for future spacecraft applications. Prior studies on experimental laboratory type Li-TiS2 cells yielded promising results in terms of cycle life and rate capability. To further assess the performance of this cell, 5 Ah engineering model cells were developed. Initially baseline cells were designed and fabricated. Each cell had 15 cathodes and 16 anodes and the ratio of anode to cathode capacity is 6:1. A solution of 1.5 molar LiAsF6 in 2Me-THF was used as the electrolyte. Cells were evaluated for their cycle life at C/1 and C/5 discharge rates and 100 percent depth of discharge. The cells were cycled between voltage limits 1.7 and 2.8 volts. The rate of charge in all cases is C/10. The results obtained indicate that cells can operate at C/10 to C/2 discharge rates and have an initial energy density of 70 Wh/kg. Cells delivered more than 100 cycles at C/2 discharge rate. The details of cell design, the test program, and the results obtained are described.

Subbarao, S.↗

NASA NTRS1987

Development of 8 cm x 8 cm silicon gridded back solar cell for space station

The 8-cm x 8-cm gridded back cell being developed for the space station photovoltaic array is described. Modeling studies show that the beginning of life power output of 1.039 W per cell may be met by several different configurations, the most promising being either a fielded or nonfielded 2 ohm-cm planar 8-mil cell. Experimental data are presented which show that a thermal alpha of 0.63 is achievable on a planar cell but at the expense of some Isc. Planar cells are found to possess short-circuit currents which are 9 percent lower than on textured cells due to the loss of near-IR radiation by transmission through the cell. Preliminary modeling shows that the efficiency of the textured cell exceeds the planar cell by approximately 9 percent at 25 C but the situation is reversed in orbit due to the high thermal alpha of textured cells (0.82). Experimental data show that the IR absorption in textured cells can be reduced by about 3 alpha points without loss of efficiency by controlling the doping concentration in the diffused layers, thus increasing the efficiency in orbit.

Lillington, D. R.↗

NASA NTRS1988

Development of ambient temperature secondary lithium cells

Subbarao, S.↗

NASA NTRS1991

Effects of proton irradiation on the performance of InP/GaAs solar cells

InP solar cells are known to be more radiation resistant than either GaAs or Si. In addition, AMO total area efficiencies approaching 19 percent were attained for InP. However, the present high substrate cost presents a barrier to the eventual widespread use of InP cells in space. In addition, if cell thinning becomes desirable, their relative fragility presents a problem. For these reasons, the NASA Lewis Research Center has initiated a program, aimed at producing thin InP cells, by heteroepitaxial deposition of InP on cheaper, more durable substrates. To date, a short term feasibility study as Spire has resulted in cells processed from InP heteroepitaxially deposited on Si substrates with an intervening thin GaAs layer (InP/GaAs/Si) and cells produced from InP deposited on GaAs (InP/GaAs). As a result of this short study efficiencies of over 7 and 9 percent were achieved for InP/GaAs/Si and InP/GaAs respectively. Although these efficiencies are low, they represent a modest and encouraging starting point for a more intensive program. Obviously, when considering economy and mechanical strength, cells processed on silicon substrates are preferred. However, although the InP/GaAs cells are not the final desirable products of this program, their properties serve to highlight several roadblocks to be overcome in producing cells with the more desirable cost and strength properties. Hence, in the present case, the properties of the InP/GaAs cells before and after irradiation by 10 MeV protons are examined. A similar study of InP/GaAs/Si cells will be reported on at a later date.

Weinberg, Irving↗