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84 records · Page 5

Targeted gene expression without a tissue-specific promoter: creating mosaic embryos using laser-induced single-cell heat shock

We have developed a method to target gene expression in the Drosophila embryo to a specific cell without having a promoter that directs expression in that particular cell. Using a digitally enhanced imaging system to identify single cells within the living embryo, we apply a heat shock to each cell individually by using a laser microbeam. A 1- to 2-min laser treatment is sufficient to induce a heat-shock response but is not lethal to the heat-shocked cells. Induction of heat shock was measured in a variety of cell types, including neurons and somatic muscles, by the expression of beta-galactosidase from an hsp26-lacZ reporter construct or by expression of a UAS target gene after induction of hsGAL4. We discuss the applicability of this technique to ectopic gene expression studies, lineage tracing, gene inactivation studies, and studies of cells in vitro. Laser heat shock is a versatile technique that can be adapted for use in a variety of research organisms and is useful for any studies in which it is desirable to express a given gene in only a distinct cell or clone of cells, either transiently or constitutively, at a time point of choice.

Non-NASA Center↗

Fluid flow releases fibroblast growth factor-2 from human aortic smooth muscle cells

This study tested the hypothesis that fluid shear stress regulates the release of fibroblast growth factor (FGF)-2 from human aortic smooth muscle cells. FGF-2 is a potent mitogen that is involved in the response to vascular injury and is expressed in a wide variety of cell types. FGF-2 is found in the cytoplasm of cells and outside cells, where it associates with extracellular proteoglycans. To test the hypothesis that shear stress regulates FGF-2 release, cells were exposed to flow, and FGF-2 amounts were measured from the conditioned medium, pericellular fraction (extracted by heparin treatment), and cell lysate. Results from the present study show that after 15 minutes of shear stress at 25 dyne/cm(2) in a parallel-plate flow system, a small but significant fraction (17%) of the total FGF-2 was released from human aortic smooth muscle cells. FGF-2 levels in the circulating medium increased 10-fold over medium from static controls (P<0.01). A 50% increase in FGF-2 content versus control (P<0.01) was found in the pericellular fraction (extracted by heparin treatment). Furthermore, a significant decrease in FGF-2 was detected in the cell lysate, indicating that FGF-2 was released from inside the cell. Cell permeability studies with fluorescent dextran were performed to examine whether transient membrane disruption caused FGF-2 release. Flow cytometry detected a 50% increase in mean fluorescence of cells exposed to 25 dyne/cm(2) versus control cells. This indicates that the observed FGF-2 release from human aortic smooth muscle cells is likely due to transient membrane disruption on initiation of flow.

Non-NASA Center↗

Carbon–concentration and carbon–climate feedbacks in CMIP6 models and their comparison to CMIP5 models

Results from the fully and biogeochemically coupled simulations in which CO 2 increases at a rate of 1 % yr –1 (1pctCO2) from its preindustrial value are analyzed to quantify the magnitude of carbon–concentration and carbon–climate feedback parameters which measure the response of ocean and terrestrial carbon pools to changes in atmospheric CO 2 concentration and the resulting change in global climate, respectively. The results are based on 11 comprehensive Earth system models from the most recent (sixth) Coupled Model Intercomparison Project (CMIP6) and compared with eight models from the fifth CMIP (CMIP5). The strength of the carbon–concentration feedback is of comparable magnitudes over land (mean ± standard deviation = 0.97 ± 0.40 PgC ppm –1 ) and ocean (0.79 ± 0.07 PgC ppm –1 ), while the carbon–climate feedback over land (–45.1 ± 50.6 PgC °C –1 ) is about 3 times larger than over ocean (–17.2 ± 5.0 PgC °C –1 ). The strength of both feedbacks is an order of magnitude more uncertain over land than over ocean as has been seen in existing studies. These values and their spread from 11 CMIP6 models have not changed significantly compared to CMIP5 models. The absolute values of feedback parameters are lower for land with models that include a representation of nitrogen cycle. The transient climate response to cumulative emissions (TCRE) from the 11 CMIP6 models considered here is 1.77 ± 0.37 °C EgC –1 and is similar to that found in CMIP5 models (1.63 ± 0.48 °C EgC –1 ) but with somewhat reduced model spread. The expressions for feedback parameters based on the fully and biogeochemically coupled configurations of the 1pctCO2 simulation are simplified when the small temperature change in the biogeochemically coupled simulation is ignored. Decomposition of the terms of these simplified expressions for the feedback parameters is used to gain insight into the reasons for differing responses among ocean and land carbon cycle models.

54 ENVIRONMENTAL SCIENCES↗

DQ Admittance Extraction for Inverter-Based Resources

The power grid industry is pushing for electromagnetic transient (EMT)-based studies for generation interconnection and planning process due to high penetrations of inverter-based resource (IBRs). Vendor-specific and site-specific black-box IBR models are preferred in those simulation studies. For small-signal analysis, measurement-based admittance models are necessary. In this paper, we demonstrate the extraction of frequency-domain dq frame IBR models. These linear models are specific to operating conditions. We demonstrate two extraction methods: frequency scan and a step response-based method. The latter relies on converting time-domain responses to Laplace-domain expressions via eigensystem realization analysis (ERA). Both can lead to dq admittance representation and the latter is time saving.

admittance model↗

DQ Admittance Extraction for Inverter-Based Resources: Preprint

The power grid industry is pushing for electro-magnetic transient (EMT)-based studies for generation interconnection and planning process due to high penetrations of inverter-based resource (IBRs). Vendor-specific and site-specific black-box IBR models are preferred in those simulation studies. For small-signal analysis, measurement-based admittance models are necessary. In this paper, we demonstrate the extraction of frequency-domain dq frame IBR models. These linear models are specific to operating conditions. We demonstrate two extraction methods: frequency scan and a step response-based method. The latter relies on converting time-domain responses to Laplace-domain expressions via eigensystem realization analysis (ERA). Both can lead to dq admittance representation and the latter is time saving.

admittance model↗

Genome reduction improves octanoic acid production in scale down bioreactors

Microorganisms in large-scale bioreactors are exposed to heterogeneous environmental conditions due to physical mixing constraints. Nutritional gradients can lead to transient expression of energetically wasteful stress responses and as a result, can reduce the titres, rates and yields of a bioprocess at larger scales. To what extent these process parameters are impacted is often unknown and therefore bioprocess scale-up comes with major risk. Designing platform strains to account for these intermittent stresses before introducing synthesis pathways is one strategy for de-risking bioprocess development. For example, Escherichia coli strain RM214 is a derivative of wild-type MG1655 that has had several genes and whole operons removed from its genome based on their metabolic cost. In this study, we engineered E. coli strain RM214 (referred to as WG02) to produce octanoic acid from glycerol in batch-flask and fed-batch bioreactor cultivations and compared it to an octanoic acid-producing E. coli MG1655 (WG01). In batch flask cultivations, the two strains performed similarly. However, in carbon limited fed-batch bioreactor cultivations, WG02 provided a greater than 22% boost to biomass compared to WG01 while maintaining similar titres of octanoic acid. Reducing the biomass accumulation of WG02 with nitrogen limited fed-batch cultivation resulted in a 16% improvement in octanoic acid titre over WG01. Finally, in a scale-down system consisting of a stirred tank reactor (representing a well-mixed zone) and plug flow reactor (representing an intermittent carbon starvation zone), WG02 again improved octanoic acid titre by almost 18% while maintaining similar biomass concentrations as WG01.

59 BASIC BIOLOGICAL SCIENCES↗

Interplanetary Coronal Mass Ejection Observed at STEREO-A, Mars, Comet 67P/Churyumov-Gerasimenko, Saturn,and New Horizons En Route to Pluto: Comparison of Its Forbush Decreases at 1.4, 3.1, and 9.9 AU

We discuss observations of the journey throughout the Solar System of a large interplanetary coronal mass ejection (ICME) that was ejected at the Sun on 14 October 2014. The ICME hit Mars on 17 October, as observed by the Mars Express, Mars Atmosphere and Volatile EvolutioN Mission (MAVEN), Mars Odyssey, and Mars Science Laboratory (MSL) missions, 44 h before the encounter of the planet with the Siding-Spring comet, for which the space weather context is provided. It reached comet 67P/Churyumov-Gerasimenko, which was perfectly aligned with the Sun and Mars at 3.1 AU, as observed by Rosetta on 22 October. The ICME was also detected by STEREO-A on 16 October at 1 AU, and by Cassini in the solar wind around Saturn on the 12 November at 9.9 AU. Fortuitously, the New Horizons spacecraft was also aligned with the direction of the ICME at 31.6 AU. We investigate whether this ICME has a nonambiguous signature at New Horizons. A potential detection of this ICME by Voyager 2 at 110-111 AU is also discussed. The multispacecraft observations allow the derivation of certain properties of the ICME, such as its large angular extension of at least 116deg, its speed as a function of distance, and its magnetic field structure at four locations from 1 to 10 AU. Observations of the speed data allow two different solar wind propagation models to be validated. Finally, we compare the Forbush decreases (transient decreases followed by gradual recoveries in the galactic cosmic ray intensity) due to the passage of this ICME at Mars, comet 67P, and Saturn.

Witasse, O.↗

Isoform-specific functions of synaptopodin-2 variants in cytoskeleton stabilization and autophagy regulation in muscle under mechanical stress

Highlights: • Muscle cells express different synaptopodin-2 isoforms with individual functions. • Reduced SYNPO2 expression results in enhanced vulnerability of myofibrils. • SYNPO2e recruites BAG3 to mechanically stressed myofibrillar areas. • SYNPO2 is an organizer and regulator of BAG3-containing proteostasis systems. • BAG3 forms functionally distinct protein complexes with different SYNPO2 isoforms. Protein homeostasis (proteostasis) in multicellular organisms depends on the maintenance of force-bearing and force-generating cellular structures. Within myofibrillar Z-discs of striated muscle, isoforms of synaptopodin-2 (SYNPO2/myopodin) act as adapter proteins that are engaged in proteostasis of the actin-crosslinking protein filamin C (FLNc) under mechanical stress. SYNPO2 directly binds F-actin, FLNc and α-actinin and thus contributes to the architectural features of the actin cytoskeleton. By its association with autophagy mediating proteins, i.e. BAG3 and VPS18, SYNPO2 is also engaged in protein quality control and helps to target mechanical unfolded and damaged FLNc for degradation. Here we show that deficiency of all SYNPO2-isoforms in myotubes leads to decreased myofibrillar stability and deregulated autophagy under mechanical stress. In addition, isoform-specific proteostasis functions were revealed. The PDZ-domain containing variant SYNPO2b and the shorter, PDZ-less isoform SYNPO2e both localize to Z-discs. Yet, SYNPO2e is less stably associated with the Z-disc than SYNPO2b, and is dynamically transferred into FLNc-containing myofibrillar lesions under mechanical stress. SYNPO2e also recruits BAG3 into these lesions via interaction with the WW domain of BAG3. Our data provide evidence for a role of myofibrillar lesions as a transient quality control compartment essential to prevent and repair contraction-induced myofibril damage in muscle and indicate an important coordinating activity for SYNPO2 therein.

60 APPLIED LIFE SCIENCES↗

Parathyroid hormone induces c-fos and c-jun messenger RNA in rat osteoblastic cells

PTH is a potent regulator of osteoblast gene expression, yet the nuclear events that mediate PTH action are poorly understood. We were interested in identifying immediate early genes which may regulate PTH-altered gene expression in the osteoblast. Therefore, we examined the effects of PTH on c-fos and c-jun gene expression in a rat osteoblastic cell line (UMR 106-01). Under control conditions, c-fos and c-jun mRNAs were present at low basal levels. After PTH treatment, c-fos mRNA abundance dramatically increased, with a maximal and transient response at 30 min. PTH also stimulated an increase in c-jun mRNA, but in a biphasic manner, with maximal levels at 30 min and 2 h. These responses were dose dependent, not altered by cotreatment with the protein synthesis inhibitor cycloheximide, and preceded PTH-induced expression of matrix metallo-proteinase-1 mRNA. Nuclear run-on assays demonstrated an increased rate of c-fos and c-jun transcription after PTH exposure. To determine the signal transduction pathways involved, second messenger analogs were tested for their ability to mimic the effects of PTH. 8-Bromo-cAMP and phorbol 12-myristate 13-acetate (PMA) caused increases in the abundance of c-fos and c-jun transcripts. Ionomycin had no effect on the expression of these genes. Pretreatment of the cells with PMA resulted in a decrease in basal c-jun expression, but did not alter the PTH-mediated increase in c-fos, c-jun, or matrix metalloproteinase-1 mRNAs.(ABSTRACT TRUNCATED AT 250 WORDS).

Non-NASA Center↗

GRO: Black hole models for gamma-ray bursts

The Burst and Transient Source Experiment (BATSE) on board the Compton Gamma Ray Observatory (CGRO) has established that the distribution of gamma-ray bursts (GRB's) is isotropic but is bound radially. This finding suggests that the bursts are either cosmological or they originate from an extended Galactic halo. The implied luminosities and the observed variability of the GRB's on time scales as short as one millisecond suggest that they originate from compact objects. We are presently studying black hole models for GRB's. Any such model must produce a non-thermal photon spectrum to agree with the observed properties. For a wide range of burst parameters the assumed bursting source consists of a non-thermal electron-positron-photon plasma of very high density. It seems possible to produce such a plasma in accretion onto black holes. In our on-going work, we are developing the kinetic theory for a non-equilibrium pair plasma. The main new features of our work are as follows: (1) We do not assume the presence of a thermal electron bath. (2) Non-thermal, high-energy pairs are allowed to have an arbitrary concentration and energy distribution. (3) There is no soft photon source in our model; initially all the photons in the plasma are either energetic X-rays or gamma-rays. (4) The initial energy distribution of the pairs as well as photons is arbitrary. (5) We collect the analytical expressions for the kinetic kernels for all relevant processes. And (6) we present a different approach to finding the time-evolution of pair and photon spectra, which is a combination of the kinetic-theory and the non-linear Monte-Carlo schemes. We have developed many Monte-Carlo programs to model various process, to take into account the time evolution, and to incorporate various physical effects which are unique to non-thermal plasmas. The hydrodynamics of fireballs in GRB's was studied before. Applying results from kinetic theory will improve our understanding of these systems.

Ruderman, Malvin↗

Rapid systemic up-regulation of genes after heat-wounding and electrical stimulation

When one leaf of a tomato plant is electrically-stimulated or heat-wounded, proteinase inhibitor genes are rapidly up-regulated in distant leaves. The identity of the systemic wound signal(s) is not yet known, but major candidates include hormones transmitted via the phloem or the xylem, the electrically-stimulated self-propagating electrical signal in the phloem (the action potential, AP), or the heat-wound-induced surge in hydraulic pressure in the xylem evoking a local change in membrane potential in adjacent living cells (the variation potential, VP). In order to discriminate between these signals we have adopted two approaches. The first approach involves applying stimuli that evoke known signals and determining whether these signals have similar effects on the "model" transcripts for proteinase inhibitors (pin) and calmodulin (cal). Here we show that a heat wound almost invariably evokes a VP, while an electrical stimulation occasionally evokes an AP, and both of these signals induce accumulation of transcripts encoding proteinase inhibitors. The second approach involves identifying the array of genes turned on by heat-wounding. To this end, we have constructed a subtractive library for heat-wounded tissue, isolated over 800 putatively up-regulated clones, and shown that all but two of the fifty that we have analyzed by Northern hybridization are, indeed, up-regulated. Here we show the early kinetics of up-regulation of three of these transcripts in the terminal (4th) leaf in response to heat-wounding the 3rd leaf, about 5 cm away. Even though these transcripts show somewhat different time courses of induction, with one peaking at 30 min, another at 15 min, and another at 5 min after flaming of a distant leaf, they all exhibit a similar pattern, i.e., a transient period of transcript accumulation preceding a period of transcript decrease, followed by a second period of transcript accumulation.

NASA Discipline Plant Biology↗

Proceedings of the 40th Lunar and Planetary Science Conference

The 40th Lunar and Planetary Science Conference included sessions on: Phoenix: Exploration of the Martian Arctic; Origin and Early Evolution of the Moon; Comet Wild 2: Mineralogy and More; Astrobiology: Meteorites, Microbes, Hydrous Habitats, and Irradiated Ices; Phoenix: Soil, Chemistry, and Habitability; Planetary Differentiation; Presolar Grains: Structures and Origins; SPECIAL SESSION: Venus Atmosphere: Venus Express and Future Missions; Mars Polar Caps: Past and Present; SPECIAL SESSION: Lunar Missions: Results from Kaguya, Chang'e-1, and Chandrayaan-1, Part I; 5 Early Nebula Processes and Models; SPECIAL SESSION: Icy Satellites of Jupiter and Saturn: Cosmic Gymnasts; Mars: Ground Ice and Climate Change; SPECIAL SESSION: Lunar Missions: Results from Kaguya, Chang'e-1, and Chandrayaan-1, Part II; Chondrite Parent-Body Processes; SPECIAL SESSION: Icy Satellites of Jupiter and Saturn: Salubrious Surfaces; SNC Meteorites; Ancient Martian Crust: Primary Mineralogy and Aqueous Alteration; SPECIAL SESSION: Messenger at Mercury: A Global Perspective on the Innermost Planet; CAIs and Chondrules: Records of Early Solar System Processes; Small Bodies: Shapes of Things to Come; Sulfur on Mars: Rocks, Soils, and Cycling Processes; Mercury: Evolution and Tectonics; Venus Geology, Volcanism, Tectonics, and Resurfacing; Asteroid-Meteorite Connections; Impacts I: Models and Experiments; Solar Wind and Genesis: Measurements and Interpretation; Mars: Aqueous Processes; Magmatic Volatiles and Eruptive Conditions of Lunar Basalts; Comparative Planetology; Interstellar Matter: Origins and Relationships; Impacts II: Craters and Ejecta Mars: Tectonics and Dynamics; Mars Analogs I: Geological; Exploring the Diversity of Lunar Lithologies with Sample Analyses and Remote Sensing; Chondrite Accretion and Early History; Science Instruments for the Mars Science Lander; . Martian Gullies: Morphology and Origins; Mars: Dunes, Dust, and Wind; Mars: Volcanism; Early Solar System Chronology; Seek Out and Explore: Upcoming and Future Missions; Mars: Early History and Impact Processes; Mars Analogs II: Chemical and Spectral; Achondrites and their Parent Bodies; and Planning for Future Exploration of the Moon The poster sessions were: Lunar Missions: Results from Kaguya, Chang'e-1, and Chandrayaan-1; LRO and LCROSS; Geophysical Analysis of the Lunar Surface and Interior; Remote Observation and Geologic Mapping of the Lunar Surface; Lunar Spectroscopy; Venus Geology, Geophysics, Mapping, and Sampling; Planetary Differentiation; Bunburra and Buzzard Coulee: Recent Meteorite Falls; Meteorites: Terrestrial History; CAIs and Chondrules: Records of Early Solar System Processes; Volatile and Organic Compounds in Chondrites; Crashing Chondrites: Impact, Shock, and Melting; Ureilite Studies; Petrology and Mineralogy of the SNC Meteorites; Martian Meteorites; Phoenix Landing Site: Perchlorate and Other Tasty Treats; Mars Polar Atmospheres and Climate Modeling; Mars Polar Investigations; Mars Near-Surface Ice; Mars: A Volatile-Rich Planet; Mars: Geochemistry and Alteration Processes; Martian Phyllosilicates: Identification, Formation, and Alteration; Astrobiology; Instrument Concepts, Systems, and Probes for Investigating Rocks and Regolith; Seeing is Believing: UV, VIS, IR, X- and Gamma-Ray Camera and Spectrometer Instruments; Up Close and Personal: In Situ Analysis with Laser-Induced Breakdown Spectroscopy and Mass Spectrometry; Jupiter and Inscrutable Io; Tantalizing Titan; Enigmatic Enceladus and Intriguing Iapetus; Icy Satellites: Cryptic Craters; Icy Satellites: Gelid Geology/Geophysics; Icy Satellites: Cool Chemistry and Spectacular Spectroscopy; Asteroids and Comets; Comet Wild 2: Mineralogy and More; Hypervelocity Impacts: Stardust Models, LDEF, and ISPE; Presolar Grains; Early Nebular Processes: Models and Isotopes; Solar Wind and Genesis: Measurements and Interpretation; Education and Public Outreach; Mercury; Pursuing Lunar Exploration; Sources and Eruptionf Lunar Basalts; Chemical and Physical Properties of the Lunar Regolith; Lunar Dust and Transient Surface Phenomena; Lunar Databases and Data Restoration; Meteoritic Samples of the Moon; Chondrites, Their Clasts, and Alteration; Achondrites: Primitive and Not So Primitive; Iron Meteorites; Meteorite Methodology; Antarctic Micrometeorites; HEDs and Vesta; Dust Formation and Transformation; Interstellar Organic Matter; Early Solar System Chronology; Comparative Planetology; Impacts I: Models and Experiments; Impacts II: Craters and Ejecta; Mars: Volcanism; Mars: Tectonics and Dynamics; Martian Stratigraphy: Understanding the Geologic History of Mars Through the Sedimentary Rock Record; Mars: Valleys and Valley Networks; Mars: Aqueous Processes in Valles Marineris and the Southern Highlands; Mars: Aqueous Geomorphology; Martian Gullies: Morphology and Origins; Mars: Dunes, Dust, and Wind; Mars: Remote Sensing; Mars: Geologic Mapping, Photogrammetry, and Cratering; Martian Mineralogy: Constraints from Missions and Laboratory Investigations; Mars Analogs: Chemical and Physical; Mars Analogs: Sulfates and Sulfides; Missions: Approaches, Architectures, Analogs, and Actualities; Not Just Skin Deep: Electron Microscopy, Heat Flow, Radar, and Seismology Instruments and Planetary Data Systems, Techniques, and Interpretation.

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