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At least 325 records · Page 18

Optimized suspension culture: the rotating-wall vessel

Suspension culture remains a popular modality, which manipulates mechanical culture conditions to maintain the specialized features of cultured cells. The rotating-wall vessel is a suspension culture vessel optimized to produce laminar flow and minimize the mechanical stresses on cell aggregates in culture. This review summarizes the engineering principles, which allow optimal suspension culture conditions to be established, and the boundary conditions, which limit this process. We suggest that to minimize mechanical damage and optimize differentiation of cultured cells, suspension culture should be performed in a solid-body rotation Couette-flow, zero-headspace culture vessel such as the rotating-wall vessel. This provides fluid dynamic operating principles characterized by 1) solid body rotation about a horizontal axis, characterized by colocalization of cells and aggregates of different sedimentation rates, optimally reduced fluid shear and turbulence, and three-dimensional spatial freedom; and 2) oxygenation by diffusion. Optimization of suspension culture is achieved by applying three tradeoffs. First, terminal velocity should be minimized by choosing microcarrier beads and culture media as close in density as possible. Next, rotation in the rotating-wall vessel induces both Coriolis and centrifugal forces, directly dependent on terminal velocity and minimized as terminal velocity is minimized. Last, mass transport of nutrients to a cell in suspension culture depends on both terminal velocity and diffusion of nutrients. In the transduction of mechanical culture conditions into cellular effects, several lines of evidence support a role for multiple molecular mechanisms. These include effects of shear stress, changes in cell cycle and cell death pathways, and upstream regulation of secondary messengers such as protein kinase C. The discipline of suspension culture needs a systematic analysis of the relationship between mechanical culture conditions and biological effects, emphasizing cellular processes important for the industrial production of biological pharmaceuticals and devices.

Non-NASA Center↗

Auxin physiology of the tomato mutant diageotropica

The tomato (Lycopersicon esculentum, Mill.) mutant diageotropica (dgt) exhibits biochemical, physiological, and morphological abnormalities that suggest the mutation may have affected a primary site of auxin perception or action. We have compared two aspects of the auxin physiology of dgt and wild-type (VFN8) seedlings: auxin transport and cellular growth parameters. The rates of basipetal indole-3-acetic acid (IAA) polar transport are identical in hypocotyl sections of the two genotypes, but dgt sections have a slightly greater capacity for IAA transport. 2,3,5-Triiodobenzoic acid and ethylene reduce transport in both mutant and wild-type sections. The kinetics of auxin uptake into VFN8 and dgt sections are nearly identical. These results make it unlikely that an altered IAA efflux carrier or IAA uptake symport are responsible for the pleiotropic effects resulting from the dgt mutation. The lack of auxin-induced cell elongation in dgt plants is not due to insufficient turgor, as the osmotic potential of dgt cell sap is less (more negative) than that of VFN8. An auxin-induced increase in wall extensibility, as measured by the Instron technique, only occurs in the VFN8 plants. These data suggest dgt hypocotyls suffer a defect in the sequence of events culminating in auxin-induced cell wall loosening.

NASA Discipline Plant Biology↗

Development of the Statocyst in Aplysia Californica: Observations on Statoconial Development - Part 1

The gravity receptor organs of gastropod molluscs, such as Aplysia californica, are bilateral paired statocysts, which contain dense statoconia within a fluid-filled cyst. Gravitational forces on the statoconia are sensed through their interaction with ciliated mechanoreceptor cells in the wall of the cyst. Larval Aplysia contain a single statolith within each statocyst; when the animals grow to a critical size, they begin producing multiple statoconia, a process that continues throughout life. The number of statoconia is highly correlated with animal weight but poorly correlated with age, indicating that stone production is related to total metabolism. The single statolith has an amorphous internal structure whereas the multiple statoconia have calcification deposited on concentric layers of membrane or matrix protein. The statolith appears to be produced within the cyst lumen but the multiple statoconia are produced within supporting cells between the receptor cells. Large adult animals have statoconia larger than those in early post-metamorphic animals which have just started producing multiple stones. The maximum statocyst diameter at which the receptor-cell cilia can suspend the statolith in the center of the cyst lumen is 45 micrometers; production of multiple stones begins when the cyst reaches this size. The mechanisms by which statoconia production is initiated and controlled are discussed.

Wiederhold, Michael L.↗

Neutrophil to Lymphocyte Ratio: A Prognostic Indicator for Astronaut Health

Short-term and long-term spaceflight missions can cause immune system dysfunction in astronauts. Recent studies indicate elevated white blood cells (WBC) and polymorphonuclear neutrophils (PMN) in astronaut blood, along with unchanged or reduced lymphocyte counts, and reduced T cell function, during short-(days) and long-(months) term spaceflight. A high PMN to lymphocyte ratio (NLR) can acts as a strong predictor of poor prognosis in cancer, and as a biomarker for subclinical inflammation in humans and chronic stress in mouse models, however, the NLR has not yet been identified as a predictor of astronaut health during spaceflight. For this, complete blood cell count data collected from astronauts and rodents that have flown for short- and long-term missions on board the International Space Station (ISS) was repurposed to determine the NLR pre-, in-, and post-flight. The results displayed that the NLR progressively increased during spaceflight in both human and mice, while a spike in the NLR was observed at post-flight landing, suggesting stress-induced factors may be involved. In addition, the ground-based chronic microgravity analog, hindlimb unloading in mice, indicated an increased NLR, along with induced myeloperoxidase expression, as measured by quantitative (q)PCR. The mechanism for increased NLR was further assessed in vitro using the NASA-developed rotating wall vessel (RWV) cell culture suspension system with human WBCs. The results indicated that simulated microgravity led to increased mature PMN counts, NLR profiles, and production of reactive oxygen species (ROS). Collectively, these studies show that an increased NLR is observed in spaceflight missions, and in chronic microgravity-analog simulation in mice, and that this effect may be potentiated by the oxidative stress response in blood cells under microgravity conditions. Furthermore, these results suggest that a disrupted NLR profile in spaceflight may further disrupt immune homeostasis, potentially causing chronic immune-mediated inflammatory diseases. Thus, we propose that the health status of astronauts during short- and long-term space missions can be monitored by their NLR profile, in addition to utilizing this measurement as a tool for interventions and countermeasure development to restore homeostatic immunity.

biomarker↗

White blood cell counting system

The design, fabrication, and tests of a prototype white blood cell counting system for use in the Skylab IMSS are presented. The counting system consists of a sample collection subsystem, sample dilution and fluid containment subsystem, and a cell counter. Preliminary test results show the sample collection and the dilution subsystems are functional and fulfill design goals. Results for the fluid containment subsystem show the handling bags cause counting errors due to: (1) adsorption of cells to the walls of the container, and (2) inadequate cleaning of the plastic bag material before fabrication. It was recommended that another bag material be selected.

Source record↗

Development of polymeric coatings for control of electro-osmotic flow in ASTP MA-011 electrophoresis technology experiment

The development of a methyl cellulose based coating system for control of electro-osmotic flow at the walls of electrophoresis cells is described. Flight electrophoresis columns were coated with this system, resulting in a flight set of six columns. In flight photography of MA-011 electrophoretic separations verified control of electro-osmotic flow.

Patterson, W. J.↗

Acoustic-optical imaging without immersion

System using membraneous end wall of Bragg cell to separate test specimen from acoustic transmission medium, operates in real time and uses readily available optical components. System can be easily set up and maintained by people with little or no training in holography.

Liu, H.↗

Effects of immobilization on spermiogenesis

The influence of immobilization stress on spermiogenesis in rats was investigated. After 96 hour immobilization, histological changes began to manifest themselves in the form of practically complete disappearance of cell population of the wall of seminiferous tubule as well as a markedly increased number of cells with pathologic mitoses. Enzymological investigations showed various changes of activity (of acid and alkaline phosphatase and nonspecific esterase) in the 24, 48, and 96 hour immobilization groups.

Meitner, E. R.↗

Comparison of composite rotor blade models: A coupled-beam analysis and an MSC/NASTRAN finite-element model

A methodology was developed for the structural analysis of composite rotor blades. This coupled-beam analysis is relatively simple to use compared with alternative analysis techniques. The beam analysis was developed for thin-wall single-cell rotor structures and includes the effects of elastic coupling. This paper demonstrates the effectiveness of the new composite-beam analysis method through comparison of its results with those of an established baseline analysis technique. The baseline analysis is an MSC/NASTRAN finite-element model built up from anisotropic shell elements. Deformations are compared for three linear static load cases of centrifugal force at design rotor speed, applied torque, and lift for an ideal rotor in hover. A D-spar designed to twist under axial loading is the subject of the analysis. Results indicate the coupled-beam analysis is well within engineering accuracy.

Hodges, Robert V.↗

Reinforced Honeycomb Panels

New honeycomb panel structure has increased strength and stiffness with little increase in weight. Some or all of walls of honeycomb cells reinforced with honeycomb panels having smaller cells, lightweight foam, or other reinforcing material. Strong, lightweight reinforced panels used in aircraft, car and truck bodies, cabinets for equipment and appliances, and buildings.

Bhat, Balakrishna T.↗

Extension-torsion coupling behavior of advanced composite tilt-rotor blades

An analytic model was developed to study the extension-bend-twist coupling behavior of an advanced composite helicopter or tilt-rotor blade. The outer surface of the blade is defined by rotating an arbitrary cross section about an initial twist axis. The cross section can be nonhomogeneous and composed of generally anisotropic materials. The model is developed based upon a three dimensional elasticity approach that is recast as a coupled two-dimensional boundary value problem defined in a curvilinear coordinate system. Displacement solutions are written in terms of known functions that represent extension, bending, and twisting and unknown functions for local cross section deformations. The unknown local deformation functions are determined by applying the principle of minimum potential energy to the discretized two-dimensional cross section. This is an application of the Ritz method, where the trial function family is the displacement field associated with a finite element (8-node isoparametric quadrilaterals) representation of the section. A computer program was written where the cross section is discretized into 8-node quadrilateral subregions. Initially the program was verified using previously published results (both three-dimensional elasticity and technical beam theory) for pretwisted isotropic bars with an elliptical cross section. In addition, solid and thin-wall multi-cell NACA-0012 airfoil sections were analyzed to illustrate the pronounced effects that pretwist, initial twist axis location, and spar location has on coupled behavior. Currently, a series of advanced composite airfoils are being modeled in order to assess how the use of laminated composite materials interacts with pretwist to alter the coupling behavior of the blade. These studies will investigate the use of different ply angle orientations and the use of symmetric versus unsymmetric laminates.

Kosmatka, J. B.↗

Aeroelastic optimization of a helicopter rotor using an efficient sensitivity analysis

To reduce oscillatory hub loads in forward flight, a structural optimization analysis of a hingeless helicopter rotor has been developed and applied. The aeroelastic analysis of the rotor is based on a finite element method in space and time, and linked with automated optimization algorithms. For the optimization analysis two types of structural representation are used: a generic stiffness-distribution and a single-cell thin-walled beam. For the first type, the design variables are nonstructural mass and its placement, chordwise center of gravity offset from the elastic axis, and stiffness. For the second type, width, height and thickness of spar are used as design variables. For the behavior constraints, frequency placement, autorotational inertia and aeroelastic stability of the blade are included. The required sensitivity derivatives are obtained using a direct analytical approach. An optimum oscillatory hub load shows a 25-77 percent reduction for the generic blade, and 30-50 percent reduction for the box-beam.

Lim, Joon W.↗

Transverse vibrations of shear-deformable beams using a general higher order theory

A general higher order theory is developed to study the static and vibrational behavior of beam structures having an arbitrary cross section that utilizes both out-of-plane shear-dependent warping and in-plane (anticlastic) deformations. The equations of motion are derived via Hamilton's principle, where the full 3D constitutive relations are used. A simplified version of the general higher-order theory is also presented for beams having an arbitrary cross section that includes out-of-plane shear deformation but assumes that stresses within the cross section and in-plane deformations are negligible. This simplified model, which is accurate for long to moderately short wavelengths, offers substantial improvements over existing higher order theories that are limited to beams with thin rectangular cross sections. The current approach will be very useful in the study of thin-wall closed-cell beams such as airfoil-type sections where the magnitude of shear-related cross-sectional warping is significant.

Kosmatka, J. B.↗

Reduction of turbulence in wind tunnels

A brief nonmathematical outline is given of modern views as to the nature of the effect of turbulence, and their bearing on the desirability of designing wind tunnels for small or large turbulence. Experiments made on a particular wind tunnel for the purpose of reducing the turbulence are described, to illustrate the influence of certain factors on the magnitude of the turbulence. Moderate changes in the size, shape, and wall thickness of cells of the honeycomb were found to have little effect. The addition of a room honeycomb at the entrance was also of little value in reducing the turbulence. The turbulence decreased with increasing distance between the honeycomb and the measuring station. A further decrease was obtained by using a large area reduction in the entrance cone, with the honeycomb at the extreme entrance end. (author)

Dryden, Hugh L↗

A Fluorescence Recovery After Photobleaching (FRAP) Technique for the Measurement of Solute Transport Across Surfactant-Laden Interfaces

The technique of Fluorescence Recovery After Photobleaching (FRAP) has been applied to the measurement of interfacial transport in two-phase systems. FRAP exploits the loss of fluorescence exhibited by certain fluorophores when over-stimulated (photobleached), so that a two-phase system, originally at equilibrium, can be perturbed without disturbing the interface by strong light from an argon-ion laser and its recovery monitored by a microscope-mounted CCD camera as it relaxes to a new equilibrium. During this relaxation, the concentration profiles of the probe solute are measured on both sides of the interface as a function of time, yielding information about the transport characteristics of the system. To minimize the size of the meniscus between the two phases, a photolithography technique is used to selectively treat the glass walls of the cell in which the phases are contained. This allows concentration measurements to be made very close to the interface and increases the sensitivity of the FRAP technique.

Browne, Edward P.↗

Screech Tones of Supersonic Jets from Bevelled Rectangular Nozzles

It is known experimentally that an imperfectly expanded rectangular jet from a thin-lip convergent nozzle emits only a single dominant screech tone. The frequency of the screech tone decreases continuously with increase in jet Mach number. However, for a supersonic jet issued from a bevelled nozzle or a convergent-divergent nozzle with straight side walls, the shock cell structure and the screech frequency pattern are fairly complicated and have not been predicted before. In this paper, it is shown that the shock cell structures of these jets can be decomposed into waveguide modes of the jet flow. The screech frequencies are related to the higher-order waveguide modes following the weakest-link screech tone theory. The measured screech frequencies are found to compare well with the predicted screech frequency curves.

Tam, Christopher K. W.↗