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At least 667 records · Page 37

Developmental changes in NMDA receptor expression in the platyfish brain

We have examined the distribution of the N-methyl-D-aspartate (NMDA) receptor in the brain of a freshwater teleost using an antibody against the R1 subunit of the receptor (NMDAR1). The primary site of localization was the nucleus olfactoretinalis (NOR), a significant gonadotropin releasing hormone (GnRH)-containing brain nucleus. The number of cells expressing NMDAR1 in this nucleus was dependent upon developmental stage, with pubescent and mature animals displaying significantly more stained cells than immature and senescent animals. This is the first reported observation of age- and maturity-related NMDA receptor association with GnRH-containing brain areas.

NASA Discipline Developmental Biology↗

Fibronectin gene expression, synthesis and accumulation during in vitro differentiation of chicken osteoblasts

A well-defined chicken osteoblast culture system(18) has been used to examine fibronectin (FN) mRNA levels, synthesis, and accumulation during in vitro differentiation and matrix mineralization. Immunofluorescent staining of cells after 6 or 18 days in culture revealed that FN was initially associated with the cell surface and in partial coalignment with cytoskeletal elements while at the latter time most FN was associated with the extracellular matrix as a ubiquitous fibrillar network. Western blot analysis of total cell-associated proteins also detected FN at all culture times. However, when results were normalized to cellular DNA, FN levels increased until 12-16 and remained relatively constant thereafter. Similarly, FN synthesis as measured by [35S]-methionine labeling, and immunoprecipitation was greatest in early cultures (culture day 3) and then declined such that synthesis decreased 60% at day 18 and 94% after 24-31 days. FN mRNA levels as measured by Northern blot analysis were well correlated with FN synthesis. These results clearly show that FN is made by primary osteoblasts during their in vitro maturation. In contrast to other osteoblast markers such as alkaline phosphatase, osteocalcin, and osteopontin, whose expression increases as cells differentiate, FN accumulates in the matrix during periods of early cell growth and attachment and then remains proportional to cell number. Results with FN differ from those obtained with collagen which continues to accumulate in the extracellular matrix during osteoblast maturation. These results are consistent with FN being important for the initial attachment of early osteoblasts or osteoblast precursors to the pericellular matrix.

NASA Discipline Musculoskeletal↗

Altered skeletal pattern of gene expression in response to spaceflight and hindlimb elevation

Spaceflight leads to osteopenia, in part by inhibiting bone formation. Using an animal model (hindlimb elevation) that simulates the weightlessness of spaceflight, we and others showed a reversible inhibition of bone formation and bone mineralization. In this study, we have measured the mRNA levels of insulin-like growth factor I (IGF-I), IGF-I receptor (IGF-IR), alkaline phosphatase, and osteocalcin in the tibiae of rats flown aboard National Aeronautics and Space Administration Shuttle Flight STS-54 and compared the results with those obtained from their ground-based controls and from the bones of hindlimb-elevated animals. Spaceflight and hindlimb elevation transiently increase the mRNA levels for IGF-I, IGF-IR, and alkaline phosphatase but decrease the mRNA levels for osteocalcin. The changes in osteocalcin and alkaline phosphatase mRNA levels are consistent with a shift toward decreased maturation, whereas the rise in IGF-I and IGF-IR mRNA levels may indicate a compensatory response to the fall in bone formation. We conclude that skeletal unloading during spaceflight or hindlimb elevation resets the pattern of gene expression in the osteoblast, giving it a less mature profile.

NASA Discipline Musculoskeletal↗

Stress-induced alterations in interferon production and class II histocompatibility antigen expression

Mild electric foot-shock has been shown to be a stressor that can alter immune responses. Male Lewis rats were exposed to one session of 16 5.0-s 1.6-mA foot-shocks. Production of interferon-gamma by splenocytes in response to concanavalin-A was decreased in spleens from the shocked rats compared to control spleens. Spleen cells from rats treated with nadolol, a peripherally acting beta-adrenergic receptor antagonist, and then shocked, showed dose-dependent attenuation of the suppression of interferon-gamma production. This suggests that catecholamines mediate shock-induced suppression of interferon-gamma production. The percentage of splenic mononuclear cells expressing class II histocompatibility (Ia) antigens on their surfaces from spleens of shocked rats was determined by flow cytometry. Significantly decreased class II positive mononuclear cells were present in the spleens of shocked rats in comparison to the spleens of control rats. This may reflect an alteration of cell trafficking or decreased production of class II antigens.

Non-NASA Center↗

Expression of gibberellin 3 beta-hydroxylase gene in a gravi-response mutant, weeping Japanese flowering cherry

Expressions of the gibberellin biosynthesis gene were investigated in a normal upright type and a gravi-response mutant, a weeping type of Japanese flowering cherry (Prunus spachiana), that is unable to support its own weight and elongates downward. A segment of the gibberellin 3 beta-hydroxylase cDNA of Prunus spachiana (Ps3ox), which is responsible for active gibberellin synthesis, was amplified by using real-time RT-PCR. The content of Ps3ox mRNA in the weeping type was much greater than that in the upright type, while the endogenous gibberellin level was much higher in the elongating zone of the weeping type. These results suggest that the amount and distribution of synthesized gibberellin regulate secondary xylem formation, and the unbalanced distribution of gibberellin affects the gravi-response of the Prunus tree.

Gravitropism/physiology↗

Pluto Express - Out of the Darkness

Pluto, discovered in 1930, is the largest of a class of primordial bodies at the edge of our solar system that have comet-like properties and remain relatively unmodified by warming from the sun. It is the only planet in the solar system not explored via robotic spacecraft. This lecture discusses the status of the Pluto Express preproject (science objectives, etc.), and its telecommunication subsystem.

Pluto Pluto Express Planetary Exploration Spacecra↗

Mars Express: The Exploration of Phobos

The origin, structure and composition of Phobos are essentially still unresolved issue, with major outcomes for understanding the origin and evolution of the Solar system, and of the Mars system. Several missions have been designed with this unique body the only target. Is Phobos a captured small body (and in this case, can we identify the degree of differentiation such a small object has undergone ?), or is there any co-generic process with Mars involved ? Are there still volatile species trapped, and if so where, and in what form ? What are the reciprocal contributions of Mars and Phobos material accreted in the other body? The number of key clues for deciphering the early solar system processes, the Mars-Phobos binary evolution, and the role Phobos could play in the future of Mars exploration (including human expeditions), are numerous, and exciting for a large community. Part of the answers are in the high resolution coverage of this object (optical and IR, for composition variation), and in its full spectral analysis, from UV to thermal IR : Mars Express is the unique mission to provide this opportunity, and orbit 756, August 22, a totally unique opportunity to approach Phobos so closely. OMEGA should provide all across Stickney, the deepest accessible material, and over more than 20 km along, a spatial resolution less than 200 m, and assess the composition (and possibly their variations) wrt silicates (mafic materials), oxides, and eventually hydrated minerals and organics. By no means we should miss this opportunity: no one would understand the rationale not to turn the instruments ON while so close, and pointing them towards the Phobos surface.

Phobos↗

Orbital Express Mission Operations Planning and Resource Management using ASPEN

As satellite equipment and mission operations become more costly, the drive to keep working equipment running with less man-power rises.Demonstrating the feasibility of autonomous satellite servicing was the main goal behind the Orbital Express (OE) mission. Planning the satellite mission operations for OE required the ability to create a plan which could be executed autonomously over variable conditions. The Automated-Scheduling and Planning Environment (ASPEN)tool, developed at the Jet Propulsion Laboratory, was used to create the schedule of events in each daily plan for the two satellites of the OE mission. This paper presents an introduction to the ASPEN tool, the constraints of the OE domain, the variable conditions that were presented within the mission, and the solution to operations that ASPEN provided. ASPEN has been used in several other domains, including research rovers, Deep Space Network scheduling research, and in flight operations for the ASE project's EO1 satellite. Related work is discussed, as are the future of ASPEN and the future of autonomous satellite servicing.

Automated Scheduling and Planning Environment (ASP↗

Orbital Express Mission Operations Planning and Resource Management using ASPEN

The Orbital Express satellite servicing demonstrator program is a DARPA program aimed at developing "a safe and cost-effective approach to autonomously service satellites in orbit". The system consists of: a) the Autonomous Space Transport Robotic Operations (ASTRO) vehicle, under development by Boeing Integrated Defense Systems, and b) a prototype modular next-generation serviceable satellite, NEXTSat, being developed by Ball Aerospace. Flexibility of ASPEN: a) Accommodate changes to procedures; b) Accommodate changes to daily losses and gains; c) Responsive re-planning; and d) Critical to success of mission planning Auto-Generation of activity models: a) Created plans quickly; b) Repetition/Re-use of models each day; and c) Guarantees the AML syntax. One SRP per day vs. Tactical team

automated planning scheduling↗

Conducting On-orbit Gene Expression Analysis on ISS: WetLab-2

WetLab-2 will enable expanded genomic research on orbit by developing tools that support in situ sample collection, processing, and analysis on ISS. This capability will reduce the time-to-results for investigators and define new pathways for discovery on the ISS National Lab. The primary objective is to develop a research platform on ISS that will facilitate real-time quantitative gene expression analysis of biological samples collected on orbit. WetLab-2 will be capable of processing multiple sample types ranging from microbial cultures to animal tissues dissected on orbit. WetLab-2 will significantly expand the analytical capabilities onboard ISS and enhance science return from ISS.

quantitative PCR↗

Towards the Gravituhedron: new expressions for NMHV gravity amplitudes

In this paper, we present new expressions for n-point NMHV tree-level gravity amplitudes. We introduce a method of factorization diagrams which is a simple graphical representation of R-invariants in Yang-Mills theory. We define the gravity analogues which we call G-invariants, and expand the NMHV gravity amplitudes in terms of these objects. We provide explicit formulas of NMHV gravity amplitudes up to eight points in terms of G-invariants, and give the general definition for any number of points. We discuss the connection to BCFW representation, special behavior under large momentum shift, the role of momentum twistors and the intricate web of spurious poles cancelation. Because of the close connection between R-invariants and the (tree-level) Amplituhedron for Yang-Mills amplitudes, we speculate that the new expansion for gravity amplitudes should correspond to the triangulation of the putative Gravituhedron geometry.

72 PHYSICS OF ELEMENTARY PARTICLES AND FIELDS↗

Heterologous expression of phosphite dehydrogenase in the chloroplast or nucleus enables phosphite utilization and genetic selection in Picochlorum spp.

Microalgae present a path to ameliorate problems associated with climate change via capture and reduction of CO2 to sustainable fuels and chemicals. Picochlorum is a genus of algae recently recognized for potential application in these regards due to its high productivity, thermotolerance, and halotolerance. Foundational genetic tools have recently been established in this genus. However, at present, genetic markers are limited, hindering genetic throughput and trait stacking approaches. To expand the suite of genetic tools and markers available for this genus, we sought to heterologously express the phosphite dehydrogenase (ptxD) gene from Pseudomonas stutzeri WM88 in both the nucleus and chloroplast of Picochlorum renovo and Picochlorum celeri. Additionally, the resultant strains allow for utilization of phosphite as a sole phosphorous source and as a nuclear and plastidial selection marker for genetic engineering. Growth analysis indicated comparable growth and composition when transgenic algae were grown in media containing phosphite as a sole phosphorus source, as compared to the conventionally used phosphate. Combined, these results expand the genetic toolbox available to the Picochlorum genus and present a potential crop protection and biocontainment strategy.

09 BIOMASS FUELS↗

Development of a novel minigenome and recombinant VSV expressing Seoul hantavirus glycoprotein-based assays to identify anti-hantavirus therapeutics

Seoul virus (SEOV) is an emerging global health threat that can cause hemorrhagic fever with renal syndrome (HFRS), which results in case fatality rates of ~2%. There are no approved treatments for SEOV infections. We developed a cell-based assay system to identify potential antiviral compounds for SEOV and generated additional assays to characterize the mode of action of any promising antivirals. Here, to test if candidate antivirals targeted SEOV glycoprotein-mediated entry, we developed a recombinant reporter vesicular stomatitis virus expressing SEOV glycoproteins. To facilitate the identification of candidate antiviral compounds targeting viral transcription/replication, we successfully generated the first reported minigenome system for SEOV. This SEOV minigenome (SEOV-MG) screening assay will also serve as a prototype assay for discovery of small molecules inhibiting replication of other hantaviruses, including Andes and Sin Nombre viruses. Ours is a proof-of-concept study in which we tested several compounds previously reported to have activity against other negative-strand RNA viruses using our newly developed hantavirus antiviral screening systems. These systems can be used under lower biocontainment conditions than those needed for infectious viruses, and identified several compounds with robust anti-SEOV activity. Our findings have important implications for the development of anti-hantavirus therapeutics.

60 APPLIED LIFE SCIENCES↗

Ablating Lgr5-expressing prostatic stromal cells activates the ERK-mediated mechanosensory signaling and disrupts prostate tissue homeostasis

The heterogeneity of the prostate stromal cells is widely appreciated but the functional implication remains incompletely understood. Using genetic lineage tracing and light-sheet imaging, we show that some fibroblast cells near the junction of the mouse proximal prostatic ducts and prostatic urethra highly express Lgr5. Genetic ablation of these anatomically restricted stromal cells, but not nonselective ablation of prostatic stromal cells, rapidly induces prostate epithelial turnover and dedifferentiation that are reversed following spontaneous restoration of the Lgr5 + stromal cells. RNA-Seq analysis implies that ablating the Lgr5 + stromal cells activates a mechanosensory response. We show that ablating the Lgr5 + stromal cells impairs the control of prostatic ductal outlet, increases prostate tissue stiffness, and activates the mitogen-activated protein kinase (MAPK). Suppressing MAPK overrides the increased epithelial proliferation. This study demonstrates that the Lgr5 + stromal cells regulate tissue homeostasis in a long-distance manner by maintaining anatomic integrity and implies that the cells near the transitional regions between organs likely control organ homeostasis by sustaining a balanced mechanoforce.

59 BASIC BIOLOGICAL SCIENCES↗

Ectopic expression of pectate lyase PtxtPL1-27 in aspen affects leaf cuticle development

Cuticle - a hydrophobic barrier of cutin and waxes covering the outer cell wall surface of plants - enables survival in terrestrial habitats. However, it is not understood how the hydrophobic cuticle precursors travel through the homogalacturonan-rich hydrophilic cell wall. To elucidate the role of homogalacturonan in cuticle development, we disrupted its integrity by overexpressing a pectate lyase, PtxtPL1-27, in aspen. PtxtPL1-27 had pleiotropic effects on shoot development, including the reduction of cuticle thickness and changes in cutin and wax composition, but the expression of cutin biosynthetic genes was little affected. Despite a reduction in homogalacturonan content in the leaves, labeling with the homogalacturonan-specific antibody JIM5 in the outer epidermal cell wall layer increased and displayed an altered pattern. Moreover, the ultrastructure of cell walls was changed concomitant with lipid accumulation. We propose that the disruption of homogalacturonan integrity affected the cutinsome-dependent transport and polymerization of cutin monomers in the cell wall.

Plant Biology↗