Search NASA⌕ Search

SEARCH · Search NASA

Results for “segmented cell”

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.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 145 records · Page 8
All sourcesNASA NTRSDOE OSTI
NASA NTRS2000

Highly ordered vertical structure of Synechococcus populations within the one-millimeter-thick photic zone of a hot spring cyanobacterial mat

A variety of contemporary techniques were used to investigate the vertical distribution of thermophilic unicellular cyanobacteria, Synechococcus spp., and their activity within the upper 1-mm-thick photic zone of the mat community found in an alkaline siliceous hot spring in Yellowstone National Park in Wyoming. Detailed measurements were made over a diel cycle at a 61 degrees C site. Net oxygenic photosynthesis measured with oxygen microelectrodes was highest within the uppermost 100- to 200-microm-thick layer until midmorning, but as the day progressed, the peak of net activity shifted to deeper layers, stabilizing at a depth of 300 microm from midday throughout the afternoon. Examination of vertical thin sections by bright-field and autofluorescence microscopy revealed the existence of different populations of Synechococcus which form discrete bands at different vertical positions. Denaturing gradient gel electrophoresis analysis of PCR-amplified 16S rRNA gene segments from horizontal cryosections obtained at 100-microm-thick vertical intervals also suggested vertical stratification of cyanobacterial, green sulfur bacterium-like, and green nonsulfur bacterium-like populations. There was no evidence of diel migration. However, image analysis of vertical thin sections revealed the presence of a narrow band of rod-shaped Synechococcus cells in which the cells assumed an upright position. These upright cells, located 400 to 800 microm below the surface, were observed only in mat samples obtained around noon. In mat samples obtained at other time points, the cells were randomly oriented throughout the mat. These combined observations reveal the existence of a highly ordered structure within the very thin photic zone of this hot spring microbial mat, consisting of morphologically similar Synechococcus populations that are likely to be differentially adapted, some co-occurring with green sulfur bacterium-like populations, and all overlying green nonsulfur bacterium-like populations.

Non-NASA Center↗
NASA NTRS2000

Molecular and biochemical evidence for the involvement of calcium/calmodulin in auxin action

The use of (35)S-labeled calmodulin (CaM) to screen a corn root cDNA expression library has led to the isolation of a CaM-binding protein, encoded by a cDNA with sequence similarity to small auxin up RNAs (SAURs), a class of early auxin-responsive genes. The cDNA designated as ZmSAUR1 (Zea mays SAURs) was expressed in Escherichia coli, and the recombinant protein was purified by CaM affinity chromatography. The CaM binding assay revealed that the recombinant protein binds to CaM in a calcium-dependent manner. Deletion analysis revealed that the CaM binding site was located at the NH(2)-terminal domain. A synthetic peptide of amino acids 20-45, corresponding to the potential CaM binding region, was used for calcium-dependent mobility shift assays. The synthetic peptide formed a stable complex with CaM only in the presence of calcium. The CaM affinity assay indicated that ZmSAUR1 binds to CaM with high affinity (K(d) approximately 15 nM) in a calcium-dependent manner. Comparison of the NH(2)-terminal portions of all of the characterized SAURs revealed that they all contain a stretch of the basic alpha-amphiphilic helix similar to the CaM binding region of ZmSAUR1. CaM binds to the two synthetic peptides from the NH(2)-terminal regions of Arabidopsis SAUR-AC1 and soybean 10A5, suggesting that this is a general phenomenon for all SAURs. Northern analysis was carried out using the total RNA isolated from auxin-treated corn coleoptile segments. ZmSAUR1 gene expression began within 10 min, increased rapidly between 10 and 60 min, and peaked around 60 min after 10 microM alpha-naphthaleneacetic acid treatment. These results indicate that ZmSAUR1 is an early auxin-responsive gene. The CaM antagonist N-(6-aminohexyl)5-chloro-1-naphthalenesulfonamide hydrochloride inhibited the auxin-induced cell elongation but not the auxin-induced expression of ZmSAUR1. This suggests that calcium/CaM do not regulate ZmSAUR1 at the transcriptional level. CaM binding to ZmSAUR1 in a calcium-dependent manner suggests that calcium/CaM regulate ZmSAUR1 at the post-translational level. Our data provide the first direct evidence for the involvement of calcium/CaM-mediated signaling in auxin-mediated signal transduction.

NASA Program Fundamental Space Biology↗
NASA NTRS1995

Regulation of insulin-like growth factor I transcription by cyclic adenosine 3',5'-monophosphate (cAMP) in fetal rat bone cells through an element within exon 1: protein kinase A-dependent control without a consensus AMP response element

Insulin-like growth factor I (IGF-I) is a locally synthesized anabolic growth factor for bone. IGF-I synthesis by primary fetal rat osteoblasts (Ob) is stimulated by agents that increase the intracellular cAMP concentration, including prostaglandin E2 (PGE2). Previous studies with Ob cultures demonstrated that PGE2 enhanced IGF-I transcription through selective use of IGF-I promoter 1, with little effect on IGF-I messenger RNA half-life. Transient transfection of Ob cultures with an array of promoter 1-luciferase reporter fusion constructs has now allowed localization of a potential cis-acting promoter element(s) responsible for cAMP-stimulated gene expression to the 5'-untranslated region (5'-UTR) of IGF-I exon 1, within a segment lacking a consensus cAMP response element. Our evidence derives from three principal observations: 1) a transfection construct containing only 122 nucleotides (nt) of promoter 1 and 328 nt of the 5'-UTR retained full PGE2-stimulated reporter expression; 2) maximal PGE2-driven reporter expression required the presence of nt 196 to 328 of exon 1 when tested within the context of IGF-I promoter 1; 3) cotransfection of IGF-I promoter-luciferase-reporter constructs with a plasmid encoding the alpha-isoform of the catalytic subunit of murine cAMP-dependent protein kinase (PKA) produced results comparable to those seen with PGE2 treatment, whereas cotransfection with a plasmid encoding a mutant regulatory subunit of PKA that cannot bind cAMP blocked PGE2-induced reporter expression. Deoxyribonuclease I footprinting of the 5'-UTR of exon 1 demonstrated protected sequences at HS3A, HS3B, and HS3D, three of six DNA-protein binding sites previously characterized with rat liver nuclear extracts. Of these three regions, only the HS3D binding site is located within the functionally identified hormonally responsive segment of IGF-I exon 1. These results directly implicate PKA in the control of IGF-I gene transcription by PGE2 and identify a segment of IGF-I exon 1 as being essential for this hormonal regulation.

Non-NASA Center↗
NASA NTRS1994

Methodologies to determine forces on bones and muscles of body segments during exercise, employing compact sensors suitable for use in crowded space vehicles

A complete description of an instrumented ergometer system, including the sensors, the data acquisition system, and the methodologies to calculate the kinematic parameters were initially developed at Tulane University. This work was continued by the PI at NASA Johnson Space Center, where a flight ergometer was instrumented and tested during a KC-135 Zero-Gravity flight. The sensors that form part of the system include EMG probes and accelerometers mounted on the subject using the ergometer, load cells to measure pedal forces, and encoders to measure position and orientation of the pedal (foot). Currently, data from the flight test is being analyzed and processed to calculate the kinematic parameters of the individual. The formulation developed during the initial months of the grant will be used for this purpose. The system's components are compact (all sensors are very small). A salient feature of the system and associated methodology to determine the kinematics is that although it uses accelerometers, position is not determined by integration. Position is determined by determining the angle of two frames of reference for which acceleration at one point is known in coordinates of both frames.

Figueroa, Fernando↗
NASA NTRS1981

Voltage gradients in solar array cavities as possible breakdown sites in spacecraft-charging-induced discharges

A possible explanation for environmentally-induced discharges on geosynchronous satellites exists in the electric fields formed in the cavities between solar cells - the small gaps formed by the cover slides, solar cells, metallic interconnects and insulating substrate. When exposed to a substorm environment, the cover slides become less negatively charged than the spacecraft ground. If the resultant electric field becomes large enough, then the interconnect could emit electrons (probably by field emission) which could be accelerated to space by the positive voltage on the covers. An experimental study was conducted using a small solar array segment in which the interconnect potential was controlled by a power supply while the cover slides were irradiated by monoenergetic electrons. It was found that discharges could be triggered when the interconnect potential became at least 500 volts negative with respect to the cover slides. Analytical modeling of satellites exposed to substorm environments indicates that such gradients are possible. Therefore, it appears that this trigger mechanism for discharges is possible.

Stevens, N. J.↗
NASA NTRS2004

Novel Image Processing Interface to Relate DSB Spatial Distribution from Immunofluorescence Foci Experiments to the State-of-the-Art Models of DNA Breakage

A recently developed software (NASARadiationTrackImage) allows a quick and automatic segmentation of foci that indicate spatial localization of specific proteins that are visualized by immunofluorescence. Of interest are the spatial and temporal distribution of foci such as gammaH2AX, a signal of the phosphorylation of a variant of the histone H2A that has been shown to correspond to DSBs, or proteins involved in DSB processing, such as ATM, Rad51, and p53, following exposures of human cells to high charge and energy (HZE) ion irradiation. Experimental data are recorded as sets of two-dimensional images in color with cells and foci of gammaH2AX, ATM, Rad51 or others shown. Different cells, levels of radiation and timing after radiation were recorded. The software allows us to calculate the number of foci per cell, overall intensity of light in foci and their spatial organization. A simple statistical model allows for testing of foci overlap (eclipse). A more complex statistical model previously known as DNAbreak simulates track structure and random chromosome geometry. It has one adjustable parameter corresponding to an average intensity of DSB creation in cubic micrometers of DNA volume per particle track or unit dose. Its limitation is the low-resolution limit both in physical space and DSB's along DNA. It works adequately on the scale of a cell and provides further insights on how the geometry of tracks and DNA affects genomic damage of the cell and subsequent repair. Future developments of the model for the description of the time evolution of DNA damage response proteins, and more robust track structure models will be discussed.

Ponomarev, A. L.↗
NASA NTRS2010

NASA Tech Briefs, April 2010

Topics covered include: Active and Passive Hybrid Sensor; Quick-Response Thermal Actuator for Use as a Heat Switch; System for Hydrogen Sensing; Method for Detecting Perlite Compaction in Large Cryogenic Tanks; Using Thin-Film Thermometers as Heaters in Thermal Control Applications; Directional Spherical Cherenkov Detector; AlGaN Ultraviolet Detectors for Dual-Band UV Detection; K-Band Traveling-Wave Tube Amplifier; Simplified Load-Following Control for a Fuel Cell System; Modified Phase-meter for a Heterodyne Laser Interferometer; Loosely Coupled GPS-Aided Inertial Navigation System for Range Safety; Sideband-Separating, Millimeter-Wave Heterodyne Receiver; Coaxial Propellant Injectors With Faceplate Annulus Control; Adaptable Diffraction Gratings With Wavefront Transformation; Optimizing a Laser Process for Making Carbon Nanotubes; Thermogravimetric Analysis of Single-Wall Carbon Nanotubes; Robotic Arm Comprising Two Bending Segments; Magnetostrictive Brake; Low-Friction, Low-Profile, High-Moment Two-Axis Joint; Foil Gas Thrust Bearings for High-Speed Turbomachinery; Miniature Multi-Axis Mechanism for Hand Controllers; Digitally Enhanced Heterodyne Interferometry; Focusing Light Beams To Improve Atomic-Vapor Optical Buffers; Landmark Detection in Orbital Images Using Salience Histograms; Efficient Bit-to-Symbol Likelihood Mappings; Capacity Maximizing Constellations; Natural-Language Parser for PBEM; Policy Process Editor for P(sup 3)BM Software; A Quality System Database; Trajectory Optimization: OTIS 4; and Computer Software Configuration Item-Specific Flight Software Image Transfer Script Generator.

Source record↗
NASA NTRS2003

Rod/Coil Block Copolyimides for Ion-Conducting Membranes

Rod/coil block copolyimides that exhibit high levels of ionic conduction can be made into diverse products, including dimensionally stable solid electrolyte membranes that function well over wide temperature ranges in fuel cells and in lithium-ion electrochemical cells. These rod/coil block copolyimides were invented to overcome the limitations of polymers now used to make such membranes. They could also be useful in other electrochemical and perhaps some optical applications, as described below. The membranes of amorphous polyethylene oxide (PEO) now used in lithium-ion cells have acceptably large ionic conductivities only at temperatures above 60 C, precluding use in what would otherwise be many potential applications at lower temperatures. PEO is difficult to process, and, except at the highest molecular weights it is not very dimensionally stable. It would be desirable to operate fuel cells at temperatures above 80 C to take advantage of better kinetics of redox reactions and to reduce contamination of catalysts. Unfortunately, proton-conduction performance of a typical perfluorosulfonic polymer membrane now used as a solid electrolyte in a fuel cell decreases with increasing temperature above 80 C because of loss of water from within the membrane. The loss of water has been attributed to the hydrophobic nature of the polymer backbone. In addition, perfluorosulfonic polymers are expensive and are not sufficiently stable for long-term use. Rod/coil block copolyimides are so named because each molecule of such a polymer comprises short polyimide rod segments alternating with flexible polyether coil segments (see figure). The rods and coils can be linear, branched, or mixtures of linear and branched. A unique feature of these polymers is that the rods and coils are highly incompatible, giving rise to a phase separation with a high degree of ordering that creates nanoscale channels in which ions can travel freely. The conduction of ions can occur in the coil phase, the rod phase, or both phases.

Meador, Mary Ann B.↗
NASA NTRS1993

The three-dimensional structure of cumulus clouds over the ocean. 1: Structural analysis

Thermal channel (channel 6, 10.4-12.5 micrometers) images of five Landsat thematic mapper cumulus scenes over the ocean are examined. These images are thresholded using the standard International Satellite Cloud Climatology Project (ISCCP) thermal threshold algorithm. The individual clouds in the cloud fields are segmented to obtain their structural statistics which include size distribution, orientation angle, horizontal aspect ratio, and perimeter-to-area (PtA) relationship. The cloud size distributions exhibit a double power law with the smaller clouds having a smaller absolute exponent. The cloud orientation angles, horizontal aspect ratios, and PtA exponents are found in good agreement with earlier studies. A technique also is developed to recognize individual cells within a cloud so that statistics of cloud cellular structure can be obtained. Cell structural statistics are computed for each cloud. Unicellular clouds are generally smaller (less than or equal to 1 km) and have smaller PtA exponents, while multicellular clouds are larger (greater than or equal to 1 km) and have larger PtA exponents. Cell structural statistics are similar to those of the smaller clouds. When each cell is approximated as a quadric surface using a linear least squares fit, most cells have the shape of a hyperboloid of one sheet, but about 15% of the cells are best modeled by a hyperboloid of two sheets. Less than 1% of the clouds are ellipsoidal. The number of cells in a cloud increases slightly faster than linearly with increasing cloud size. The mean nearest neighbor distance between cells in a cloud, however, appears to increase linearly with increasing cloud size and to reach a maximum when the cloud effective diameter is about 10 km; then it decreases with increasing cloud size. Sensitivity studies of threshold and lapse rate show that neither has a significant impact upon the results. A goodness-of-fit ratio is used to provide a quantitative measure of the individual cloud results. Significantly improved results are obtained after applying a smoothing operator, suggesting the eliminating subresolution scale variations with higher spatial resolution may yield even better shape analyses.

Kuo, Kwo-Sen↗
NASA NTRS2000

Molluscan engrailed expression, serial organization, and shell evolution

Whether the serial features found in some molluscs are ancestral or derived is considered controversial. Here, in situ hybridization and antibody studies show iterated engrailed-gene expression in transverse rows of ectodermal cells bounding plate field development and spicule formation in the chiton, Lepidochitona cavema, as well as in cells surrounding the valves and in the early development of the shell hinge in the clam, Transennella tantilla. Ectodermal expression of engrailed is associated with skeletogenesis across a range of bilaterian phyla, suggesting a single evolutionary origin of invertebrate skeletons. The shared ancestry of bilaterian-invertebrate skeletons may help explain the sudden appearance of shelly fossils in the Cambrian. Our interpretation departs from the consideration of canonical metameres or segments as units of evolutionary analysis. In this interpretation, the shared ancestry of engrailed-gene function in the terminal/posterior addition of serially repeated elements during development explains the iterative expression of engrailed genes in a range of metazoan body plans.

Non-NASA Center↗
NASA NTRS1988

STS-26/Discovery Preparations for Launch

This NASA Kennedy Space Center two-part video release is comprised of footage covering STS-26 launch preparations from the arrival of the Tracking and Data Relay Satellite (TDRS) at the Orbiter Processing Facility (OPF) to the lift and mate of the external tanks. The STS-26 flight crew include: Frederick H. (Rick) Hauck, mission commander; Richard O. Covey, pilot; John M. (Mike) Lounge, mission specialist; David C. Hilmers, mission specialist; and George D. (Pinky) Nelson, mission specialist. The primary payload of STS-26 is the TDRS while the secondary payloads include the Physical Vapor Transport of Organic Solids (PVTOS); Protein Crystal Growth (PCG); Infrared Communications Flight Experiment (IRCFE); Aggregation of Red Blood Cells (ARC); Isoelectric Focusing Experiment (IFE); Mesoscale Lightning Experiment (MLE); Phase Partitioning Experiment (PPE); Earth-Limb Radiance Experiment (ELRAD); Automated Directional Solidification Furnace (ADSF) and two Shuttle Student Involvement Program (SSIP) experiments. Launch preparation footage includes flight crew arrival at KSC, rollout of Discovery to Pad B, OV-103 Discovery power-up, main engine unpacking and installation, solid rocket boosters' arrival prep and stacking, and aft skirt to aft segment mating.

Source record↗
NASA NTRS2010

International Space Station USOS Potable Water Dispenser On-Orbit Functionality Versus Design

The International Space Station (ISS) currently provides potable water dispensing for rehydrating crewmember food and drinking packages. There is one system located in the United States On-orbit Segment (USOS) and one system in the Russian Segment. Shuttle mission STS-126 delivered the USOS Potable Water Dispenser (PWD) to ISS on ULF2; subsequent activation occurred on November 2008. The PWD is capable of supporting an ISS crew of six, but nominally supplies only half this crew size. The PWD design provides incremental quantities of hot and ambient temperature potable water to US food and beverage packages. PWD receives iodinated water from the US Water Recovery System (WRS) Fuel Cell Water Bus, which feeds from the Water Processing Assembly (WPA). The PWD removes the biocidal iodine to make the water potable prior to dispensing. A heater assembly contained within the unit supplies up to 2.0 L of hot water (65 to 93 ?C) every 30 min. During a single meal, this quantity of water supports three to four crewmembers? food rehydration and beverages. The unit design has a functional life expectancy of 10 years, with replacement of limited life items, such as filters. To date, the PWD on-orbit performance is acceptable. Since activation of the PWD, there were several differences between on-orbit functionality and expected performance of hardware design. The comparison of on-orbit functionality to performance of hardware design is discussed for the following key areas: 1) microbial contamination, 2) no-dispense and water leakage scenarios, and 3) under-dispense scenarios.

Toon, Katherine P.↗
NASA NTRS2019

Vestibulo-Spinal Pathways in Tetrapods: Chapter - 6.26

The vestibulospinal system provides the spinal motor circuits controlling head/neck and limb movements and body posture with rapid reflex adjustments to maintain equilibrium and stability and with a continuous essential excitatory drive, called tonus, to enhance reactive responses to perturbations that force the animal off normal posture. The sensory signals to these reflex circuits originate from hair cells in the inner ear of otolith structures, namely the utricle and saccule, that transduce inertial acceleration and orientation of the head with respect to gravity and in the three orthogonally arranged semicircular canals that transduce angular head rotation. The principal vestibulospinal pathways are 1) the medial vestibulospinal tract that descends in the ventromedial funiculus and innervates inter- and motoneurons located mainly in lamina VII, VIII, and dorsomedial IX throughout the cervical segments; and 2) the lateral vestibulospinal tracts that course in the lateral to ventrolateral funiculi and are distinguished by two divisions: i) a cervical-projecting tract that overlaps many of the targets of medial vestibulospinal tract neurons including the motoneurons in ventromedial IX and also contributes to reflex control of shoulder and forelimb (arm) muscles; and ii) a lumbosacral-projecting tract that provides a rapid input to maintain stable posture and reflex control of the lower body. A striking observation in understanding the functional organization of this sensory-motor system is both that the driving sensory input can be dynamically modified by the behavioral context in which the sensation is made and that it remains able to quickly respond to an external force during self-generated head movements. The structural basis for vestibulospinal inputs to spinal motor control circuits in quadrupeds and bipeds rely in part on the animal's need for coordination between fore- and hind-limb reflex movements. Understanding the sensory-to-motor transformations in the diverse species rely on the correlations of the conserved and unique species behavior, morphology and physiologic function.

Spinal Cord↗
NASA NTRS2006

Numerical Evaluation of Mode 1 Stress Intensity Factor as a Function of Material Orientation For BX-265 Foam Insulation Material

Foam; a cellular material, is found all around us. Bone and cork are examples of biological cell materials. Many forms of man-made foam have found practical applications as insulating materials. NASA uses the BX-265 foam insulation material on the external tank (ET) for the Space Shuttle. This is a type of Spray-on Foam Insulation (SOFI), similar to the material used to insulate attics in residential construction. This foam material is a good insulator and is very lightweight, making it suitable for space applications. Breakup of segments of this foam insulation on the shuttle ET impacting the shuttle thermal protection tiles during liftoff is believed to have caused the space shuttle Columbia failure during re-entry. NASA engineers are very interested in understanding the processes that govern the breakup/fracture of this complex material from the shuttle ET. The foam is anisotropic in nature and the required stress and fracture mechanics analysis must include the effects of the direction dependence on material properties. Material testing at NASA MSFC has indicated that the foam can be modeled as a transversely isotropic material. As a first step toward understanding the fracture mechanics of this material, we present a general theoretical and numerical framework for computing stress intensity factors (SIFs), under mixed-mode loading conditions, taking into account the material anisotropy. We present mode I SIFs for middle tension - M(T) - test specimens, using 3D finite element stress analysis (ANSYS) and FRANC3D fracture analysis software, developed by the Cornel1 Fracture Group. Mode I SIF values are presented for a range of foam material orientations. Also, NASA has recorded the failure load for various M(T) specimens. For a linear analysis, the mode I SIF will scale with the far-field load. This allows us to numerically estimate the mode I fracture toughness for this material. The results represent a quantitative basis for evaluating the strength and fracture properties of anisotropic foam insulation material.

Knudsen, Erik↗
NASA NTRS2009

International Space Station USOS Potable Water Dispenser On-Orbit Functionality vs Design

The International Space Station (ISS) currently provides potable water dispensing for rehydrating crewmembers food and drinking packages with one system located in the United States On-orbit Segment (USOS) and one system in the Russian Segment. The USOS Potable Water Dispenser (PWD) was delivered to ISS on ULF2, Shuttle Mission STS-126, and was subsequently activated in November 2008. The PWD activation on ISS is capable of supporting an ISS crew of six but nominally supplies only half the crew. The PWD is designed to provide incremental quantities of hot and ambient temperature potable water to US style food packages. PWD receives iodinated water from the US Laboratory Fuel Cell Water Bus, which is fed from the Water Processing Assembly (WPA). The PWD removes the biocidal iodine to make the water potable prior to dispensing. A heater assembly contained within the unit supplies up to 2.0 liters of hot water (65 to 93oC) every thirty minutes. This quantity supports three to four crewmembers to rehydrate their food and beverages from this location during a single meal. The unit is designed to remain functional for up to ten years with replacement of limited life items such as filters. To date, the PWD on-orbit performance has been acceptable. Since activation of the PWD, there have been several differences between on-orbit functionality and expected performance of hardware design. The comparison of on-orbit functionality to performance of hardware design is outlined for the following key areas: microbiology, PWD to food package water leakage, no-dispense scenarios, under-dispense scenarios, and crewmember feedback on actual on-orbit use.

Toon, Katherine P.↗
NASA NTRS1998

1,25-Dihydroxyvitamin D3 inhibition of col1a1 promoter expression in calvariae from neonatal transgenic mice

We studied the effect of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) on organ cultures of transgenic mouse calvariae containing segments of the Col1a1 promoter extending to -3518, -2297, -1997, -1794, -1763, and -1719 bp upstream of the transcription start site fused to the chloramphenicol acetyltransferase (CAT) reporter gene. 1,25(OH)2D3 had a dose-dependent inhibitory effect on the expression of the -3518 bp promoter construct (ColCAT3.6), with maximal inhibition of about 50% at 10 nM. This level of inhibition was consistent with the previously observed effect on the endogenous Col1a1 gene in bone cell models. All of the shorter constructs were also inhibited by 10 nM 1,25(OH)2D3, suggesting that the sequences required for 1, 25(OH)2D3 inhibition are downstream of -1719 bp. The inhibitory effect of 1,25(OH)2D3 on transgene mRNA was maintained in the presence of the protein synthesis inhibitor cycloheximide, suggesting that the inhibitory effect on Col1a1 gene transcription does not require de novo protein synthesis. We also examined the in vivo effect of 1,25(OH)2D3 treatment of transgenic mice on ColCAT activity, and found that 48 h treatment caused a dose-dependent inhibition of CAT activity in calvariae comparable to that observed in organ cultures. In conclusion, we demonstrated that 1,25(OH)2D3 inhibits Col1A1 promoter activity in transgenic mouse calvariae, both in vivo and in vitro. The results indicate that there is a 1, 25(OH)2D3 responsive element downstream of -1719 bp. The inhibitory effect does not require new protein synthesis.

NASA Discipline Cell Biology↗
NASA NTRS1986

Cadmium-induced accumulation of putrescine in oat and bean leaves

The effects of Cd2+ on putrescine (Put), spermidine (Spd), and spermine (Spm) titers were studied in oat and bean leaves. Treatment with Cd2+ for up to 16 hours in the light or dark resulted in a large increase in Put titer, but had little or no effect on Spd or Spm. The activity of arginine decarboxylase (ADC) followed the pattern of Put accumulation, and experiments with alpha-difluoromethylarginine established that ADC was the enzyme responsible for Put increase. Concentrations of Cd2+ as low as 10 micromolar increased Put titer in oat segments. In bean leaves, there was a Cd(2+)-induced accumulation of Put in the free and soluble conjugated fractions, but not in the insoluble fraction. This suggests a rapid exchange between Put that exists in the free form and Put found in acid soluble conjugate forms. It is concluded that Cd2+ can act like certain other stresses (K+ and Mg2+ deficiency, excess NH4+, low pH, salinity, osmotic stress, wilting) to induce substantial increases in Put in plant cells.

NASA Discipline Plant Biology↗
NASA NTRS1992

Zuotin, a putative Z-DNA binding protein in Saccharomyces cerevisiae

A putative Z-DNA binding protein, named zuotin, was purified from a yeast nuclear extract by means of a Z-DNA binding assay using [32P]poly(dG-m5dC) and [32P]oligo(dG-Br5dC)22 in the presence of B-DNA competitor. Poly(dG-Br5dC) in the Z-form competed well for the binding of a zuotin containing fraction, but salmon sperm DNA, poly(dG-dC) and poly(dA-dT) were not effective. Negatively supercoiled plasmid pUC19 did not compete, whereas an otherwise identical plasmid pUC19(CG), which contained a (dG-dC)7 segment in the Z-form was an excellent competitor. A Southwestern blot using [32P]poly(dG-m5dC) as a probe in the presence of MgCl2 identified a protein having a molecular weight of 51 kDa. The 51 kDa zuotin was partially sequenced at the N-terminal and the gene, ZUO1, was cloned, sequenced and expressed in Escherichia coli; the expressed zuotin showed similar Z-DNA binding activity, but with lower affinity than zuotin that had been partially purified from yeast. Zuotin was deduced to have a number of potential phosphorylation sites including two CDC28 (homologous to the human and Schizosaccharomyces pombe cdc2) phosphorylation sites. The hexapeptide motif KYHPDK was found in zuotin as well as in several yeast proteins, DnaJ of E.coli, csp29 and csp32 proteins of Drosophila and the small t and large T antigens of the polyoma virus. A 60 amino acid segment of zuotin has similarity to several histone H1 sequences. Disruption of ZUO1 in yeast resulted in a slow growth phenotype.

NASA Discipline Exobiology↗