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At least 19 records

Altering the cellular mechanical force balance results in integrated changes in cell, cytoskeletal and nuclear shape

Studies were carried out with capillary endothelial cells cultured on fibronectin (FN)-coated dishes in order to analyze the mechanism of cell and nuclear shape control by extracellular matrix (ECM). To examine the role of the cytoskeleton in shape determination independent of changes in transmembrane osmotic pressure, membranes of adherent cells were permeabilized with saponin (25 micrograms/ml) using a buffer that maintains the functional integrity of contractile microfilaments. Real-time videomicroscopic studies revealed that addition of 250 microM ATP resulted in time-dependent retraction and rounding of permeabilized cells and nuclei in a manner similar to that observed in intact living cells following detachment using trypsin-EDTA. Computerized image analysis confirmed that permeabilized cells remained essentially rigid in the absence of ATP and that retraction was stimulated in a dose-dependent manner as the concentration of ATP was raised from 10 to 250 microM. Maximal rounding occurred by 30 min with projected cell and nuclear areas being reduced by 69 and 41%, respectively. ATP-induced rounding was also accompanied by a redistribution of microfilaments resulting in formation of a dense net of F-actin surrounding retracted nuclei. Importantly, ATP-stimulated changes in cell, cytoskeletal, and nuclear form were prevented in permeabilized cells using a synthetic myosin peptide (IRICRKG) that has been previously shown to inhibit actomyosin filament sliding in muscle. In contrast, both the rate and extent of cell and nuclear rounding were increased in permeabilized cells exposed to ATP when the soluble FN peptide, GRGDSP, was used to dislodge immobilized FN from cell surface integrin receptors.(ABSTRACT TRUNCATED AT 250 WORDS).

Non-NASA Center

Three-Dimensional Analysis of Nuclear Size, Shape and Displacement in Clover Root Cap Statocytes from Space and a Clinostat

Under normal (l-g) conditions the statocytes of root caps have a characteristic polarity with the nucleus in tight association with the proximal cell wall; but, in altered gravity environments including microgravity (mu-g) and the clinostat (c-g) movement of the nucleus away from the proximal cell wall is not uncommon. To further understand the cause of gravity-dependent nuclear displacement in statocytes, three-dimensional cell reconstruction techniques were used to precisely measure the volumes, shapes, and positions of nuclei in white clover (Trifolium repens) flown in space and rotated on a clinostat. Seeds were germinated and grown for 72 hours aboard the Space Shuttle (STS-63) in the Fluid Processing Apparatus (BioServe Space Technologies, Univ. of Colorado, Boulder). Clinorotation experiments were performed on a two-axis clinostat (BioServe). Computer reconstruction of selected groups of statocytes were made from serial sections (0.5 microns thick) using the ROSS (Reconstruction Of Serial Sections) software package (Biocomputation Center, NASA Ames Research Center). Nuclei were significantly displaced from the tops of cells in mu-g (4.2 +/- 1.0 microns) and c-g (4.9 +/- 1.4 microns) when compared to l-g controls (3.4 +/- 0.8 gm); but, nuclear volume (113 +/- 36 cu microns, 127 +/- 32 cu microns and 125 +/- 28 cu microns for l-g, mu-g and c-g respectively) and the ratio of nuclear volume to cell volume (4.310.7%, 4.211.0% and 4.911.4% respectively) were not significantly dependent on gravity treatment (ANOVA; alpha = 0.05). Three-dimensional analysis of nuclear shape and proximity to the cell wall, however, showed that nuclei from l-g controls appeared ellipsoidal while those from space and the clinostat were more spherically shaped. This change in nuclear shape may be responsible for its displacement under altered gravity conditions. Since the cytoskeleton is known to affect nuclear polarity in root cap statocytes, those same cytoskeletal elements could also control nuclear shape. This alteration in nuclear shape and position in mu-g and c-g when compared to l-g may lead to functional differences in the gravity signaling systems of plants subjected to altered gravity environments.

Smith, J.D.

DNA Break Clustering as a Predictor of Cell Death across Various Radiation Qualities: Influence of Cell Size, Cell Asymmetry, and Beam Orientation

Cosmic radiation, composed of high charge and energy (HZE) particles, causes cellular DNA damage that can result in cell death or mutation that can evolve into cancer. In this work, a cell death model is applied to several cell lines exposed to HZE ions spanning a broad range of linear energy transfer (LET) values. We hypothesize that chromatin movement leads to the clustering of multiple double strand breaks (DSB) within one radiation-induced foci (RIF). The survival probability of a cell population is determined by averaging the survival probabilities of individual cells, which is function of the number of pairwise DSB interactions within RIF. The simulation code RITCARD was used to compute DSB. Two clustering approaches were applied to determine the number of RIF per cell. RITCARD outputs were combined with experimental data from four normal human cell lines to derive the model parameters and expand its predictions in response to ions with LET ranging from ∼0.2keV/μmto∼3000keV/μm. Spherical and ellipsoidal nuclear shapes and two ion beam orientations were modeled to assess the impact of geometrical properties on cell death. The calculated average number of RIF per cell reproduces the saturation trend for high doses and high-LET values that is usually experimentally observed. The cell survival model generates the recognizable bell shape of LET dependence for the relative biological effectiveness (RBE). At low LET, smaller nuclei have lower survival due to increased DNA density and DSB clustering. At high LET, nuclei with a smaller irradiation area either because of a smaller size or a change in beam orientation have a higher survival rate due to a change in the distribution of DSB/RIF per cell. If confirmed experimentally, the geometric characteristics of cells would become a significant factor in predicting radiation-induced biological effects.

cell survival

Shapes and stability of algebraic nuclear models

A generalization of the procedure to study shapes and stability of algebraic nuclear models introduced by Gilmore is presented. One calculates the expectation value of the Hamiltonian with respect to the coherent states of the algebraic structure of the system. Then equilibrium configurations of the resulting energy surface, which depends in general on state variables and a set of parameters, are classified through the Catastrophe theory. For one- and two-body interactions in the Hamiltonian of the interacting Boson model-1, the critical points are organized through the Cusp catastrophe. As an example, we apply this Separatrix to describe the energy surfaces associated to the Rutenium and Samarium isotopes.

Lopez-Moreno, Enrique

Neutron star crusts

We calculate properties of neutron star matter at subnuclear densities using an improved nuclear Hamiltonian. Nuclei disappear and the matter becomes uniform at a density of about 0.6n(s), where n(s) of about 0.16/cu fm is the saturation density of nuclear matter. As a consequence, the mass of matter in the crusts of neutron stars is only about half as large as previously estimated. In about half of that crustal mass, nuclear matter occurs in shapes very different from the roughly spherical nuclei familiar at lower densities. The thinner crust and the unusual nuclear shape have important consequences for theories of the rotational and thermal evolution of neutron stars, especialy theories of glitches.

Lorenz, C. P.

Devescovinid trichomonad with axostyle-based rotary motor ("Rubberneckia"): taxonomic assignment as Caduceia versatilis sp. nov

An amitochondriate trichomonad cell of the family Devescovinidae (Class Parabasalia), helped demonstrate the fluid model of lipoprotein cell membranes. This wood-ingesting symbiont in the hindgut of the dry wood-eating termite Cryptotermes cavifrons is informally known to cell biologists as "Rubberneckia". As the microtubular axo-style complex generates force causing clockwise movement of the entire anterior portion of the cell at the shear zone the protist displays "head" rotation. Studies by phase contrast and videomicroscopy of live cells, of whole mounts by scanning, and thin sections by transmission electron microscopy extend the observations of Tamm and Tamm [24-26] and Tamm [19-23]. Habitat, cell shape, size, nuclear features, parabasal apparatus and other morphological details permit the assignment of "Rubberneckia" to Kirby's cosmopolitan genus Caduceia. This large-sized devescovinid has distinctive parabasal gyres, an axostylar rotary, motor, and regularly-associated nonflagellated, fusiform and flagellated rod epibiotic surface bacteria. In addition to regularly aligned epibionts intranuclear and endocytoplasmic bacteria are abundant and hydrogenosomes are Present. "Rubberneckia" is compared here to the other seven species of Caduceia. Since it is clearly sufficiently distinctive to warrant new species status, we named it C. versatilis.

Non-NASA Center

Comments on comet shapes and aggregation processes

An important question for a comet mission is whether comet nuclei preserve information clarifying aggregation processes of planetary matter. New observational evidence shows that Trojan asteroids, as a group, display a higher fraction of highly-elongated objects than the belt. More recently evidence has accumulated that comet nuclei, as a group, also display highly-elongated shapes at macro-scale. This evidence comes from the several comets whose nuclear lightcurves or shapes have been well studied. Trojans and comet nuclei share other properties. Both groups have extremely low albedos and reddish-to neutral-black colors typical of asteroids of spectral class D, P, and C. Both groups may have had relatively low collision frequencies. An important problem to resolve with spacecraft imaging is whether these elongated shapes are primordial, or due to evolution of the objects. Two hypotheses that might be tested by a combination of global-scale and close-up imaging from various directions are: (1) The irregular shapes are primordial and related to the fact that these bodies have had lower collision frequencies than belt asteroids; or (2) The irregular shapes may be due to volatile loss.

Hartmann, William K.

Effect of filtering on noise and timing in nuclear particle detection

The relations between signal shaping, noise, and timing resolution in nuclear particle detection systems are examined in this paper. Mathematical analysis of five commonly used types of filters leads to numerical results for system noise components arising from sources in the preamplifier, detector, and bias resistors. These results are presented in the form of design curves for the various filters, showing the dependence of each output noise component on both the magnitude of the noise sources and the filter time constants. Three of the filters considered are simple RC types; the remaining two are combinations of delay lines and RC elements. The quantitative calculations specifically assume a silicon detector and a field-effect-transistor preamplifier. Those filters that exhibit bipolar step response are analyzed in terms of crossover timing resolution, based on the earlier noise calculations. Again, the results are summarized in sets of design curves for each filter. Finally, the comparative performance of the various filter types is discussed, and specific examples are given.

Gary, S. A.

Nuclear pumped laser research at the Jet Propulsion Laboratory

Using a partially nuclear excited xenon flashlamp to pump an iodine laser, laser pulse shapes were analyzed with and without nuclear flashlamp augmentation. The pulse shapes indicate that the deposition of nuclear energy is equally as effective as electrical energy deposition in producing laser pulse energy output. The amplification of the E-beam pumped CF3I was measured at pressures of several atmospheres. Preliminary data shows that, for a part of the iodine laser pulse, amplification of almost a factor of two is measured. This measurement indicates that the gain in an E-beam pumped CF3I is an order of magnitude greater than in the coaxial laser tube.

Russell, G. R.

Abrasion cross sections for Ne-20 projectiles at 2.1 GeV/nucleon

Utilizing eikonal scattering theory, an optical model potential approximation to the exact nucleus-nucleus multiple scattering series is used in an abrasion-ablation collision formalism to predict abrasion cross sections for relativistic Ne-20 projectile nuclei. Excellent agreement with recent experimental abrasion results is obtained. The sensitivity of the abrasion predictions to Pauli exclusion principle correlation effects and to the assumed shape of the nuclear single-particle density distribution is also demonstrated.

Townsend, L. W.

CCD-photometry of comets at large heliocentric distances

CCD imaging and time series photometry are used to determine the state of activity, nuclear properties and eventually the rotational motion of cometary nuclei. Cometary activity at large heliocentric distances and mantle evolution are not yet fully understood. Results of observations carried out at the 2.1 telescope on Kitt Peak April 10-12 and May 15-16, 1991 are discussed. Color values and color-color diagrams are presented for several comets and asteroids. Estimations of nuclear radii and shapes are given.

Mueller, Beatrice E. A.

Quasi-projection-operator calculation of inelastic resonances in He/-/ and Li

Feshbach resonances of electrons incident of He and Li(+) are calculated below the 3(S-3) target threshold by means of the inelastic quasi-projection-operator technique of Temkin and Bhatia (1972). A configuration-interaction-type wave function is employed with up to 46 configurations. The calculations reveal, with reasonable accuracy, all presently observed Feshbach resonances. The method does not reveal shape resonances, but, for isoelectronic systems with greater nuclear charge, it is argued that shape resonance should become less important. This is already apparent in the case of Li, for which the known resonances are calculated and higher resonances, for which there are as yet no experimental data, are predicted.

Wakid, S.

Cell shape, cytoskeletal mechanics, and cell cycle control in angiogenesis

Capillary endothelial cells can be switched between growth and differentiation by altering cell-extracellular matrix interactions and thereby, modulating cell shape. Studies were carried out to determine when cell shape exerts its growth-regulatory influence during cell cycle progression and to explore the role of cytoskeletal structure and mechanics in this control mechanism. When G0-synchronized cells were cultured in basic fibroblast growth factor (FGF)-containing defined medium on dishes coated with increasing densities of fibronectin or a synthetic integrin ligand (RGD-containing peptide), cell spreading, nuclear extension, and DNA synthesis all increased in parallel. To determine the minimum time cells must be adherent and spread on extracellular matrix (ECM) to gain entry into S phase, cells were removed with trypsin or induced to retract using cytochalasin D at different times after plating. Both approaches revealed that cells must remain extended for approximately 12-15 h and hence, most of G1, in order to enter S phase. After this restriction point was passed, normally 'anchorage-dependent' endothelial cells turned on DNA synthesis even when round and in suspension. The importance of actin-containing microfilaments in shape-dependent growth control was confirmed by culturing cells in the presence of cytochalasin D (25-1000 ng ml-1): dose-dependent inhibition of cell spreading, nuclear extension, and DNA synthesis resulted. In contrast, induction of microtubule disassembly using nocodazole had little effect on cell or nuclear spreading and only partially inhibited DNA synthesis. Interestingly, combination of nocodazole with a suboptimal dose of cytochalasin D (100 ng ml-1) resulted in potent inhibition of both spreading and growth, suggesting that microtubules are redundant structural elements which can provide critical load-bearing functions when microfilaments are partially compromised. Similar synergism between nocodazole and cytochalasin D was observed when cytoskeletal stiffness was measured directly in living cells using magnetic twisting cytometry. These results emphasize the importance of matrix-dependent changes in cell and nuclear shape as well as higher order structural interactions between different cytoskeletal filament systems for control of capillary cell growth during angiogenesis.

Non-NASA Center

Observations of disk-shaped bodies in free flight at terminal velocity

Ten disk-shaped models of a proposed nuclear heat source module were released from an aircraft and observed by radar. The initial launch attitude, spin rate, and mass of the models were varied. Significant differences were observed in the mode of flight and terminal velocity among models of different mass and launch attitudes. The data were analyzed to yield lift and drag coefficients as a function of Reynolds number. The total sea-level velocity of the models was found to be well correlated as a function of mass per unit frontal area. The demonstrated terminal velocity of the modular disk heat source, about 27 m/sec for this specific design, is only 33% of that of existing heat source designs.

Vorreiter, J. W.

The shape of the future long-haul transport airplane

Long-haul transport plane configurations are discussed that are considered technically feasible within a practical level of near-term technology. Supersonic and transonic aircraft are considered, as well as different types of subsonic transports. It is indicated that superior second-generation supersonic transports are quite feasible technically; in particular, it is shown that problems of fuel consumption, efficient overland flight, and questions on noise and nitric oxides have straightforward engineering solutions. Transonic aircraft are practical but would have to have a different geometry than subsonic aircraft to provide the versatility necessary to compete at one end of their capability with supersonic transports and at the other end, for shorter ranges, with subsonic transports. The shape of subsonic transports is not expected to change noticeably, although recent advances of airfoil technology imply that new subsonic transports would have wings with higher aspect ratios and somewhat less sweep. But large specialized freighter aircraft and very long range laminar flow control or nuclear-powered aircraft with radically new shapes are possible.

Clay, C. W.