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At least 109 records · Page 6

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NASA NTRS1999

Tissue-engineered human bioartificial muscles expressing a foreign recombinant protein for gene therapy

Murine skeletal muscle cells transduced with foreign genes and tissue engineered in vitro into bioartificial muscles (BAMs) are capable of long-term delivery of soluble growth factors when implanted into syngeneic mice (Vandenburgh et al., 1996b). With the goal of developing a therapeutic cell-based protein delivery system for humans, similar genetic tissue-engineering techniques were designed for human skeletal muscle stem cells. Stem cell myoblasts were isolated, cloned, and expanded in vitro from biopsied healthy adult (mean age, 42 +/- 2 years), and elderly congestive heart failure patient (mean age, 76 +/- 1 years) skeletal muscle. Total cell yield varied widely between biopsies (50 to 672 per 100 mg of tissue, N = 10), but was not significantly different between the two patient groups. Percent myoblasts per biopsy (73 +/- 6%), number of myoblast doublings prior to senescence in vitro (37 +/- 2), and myoblast doubling time (27 +/- 1 hr) were also not significantly different between the two patient groups. Fusion kinetics of the myoblasts were similar for the two groups after 20-22 doublings (74 +/- 2% myoblast fusion) when the biopsy samples had been expanded to 1 to 2 billion muscle cells, a number acceptable for human gene therapy use. The myoblasts from the two groups could be equally transduced ex vivo with replication-deficient retroviral expression vectors to secrete 0.5 to 2 microg of a foreign protein (recombinant human growth hormone, rhGH)/10(6) cells/day, and tissue engineered into human BAMs containing parallel arrays of differentiated, postmitotic myofibers. This work suggests that autologous human skeletal myoblasts from a potential patient population can be isolated, genetically modified to secrete foreign proteins, and tissue engineered into implantable living protein secretory devices for therapeutic use.

Non-NASA Center↗

NASA NTRS1998

Attenuation of skeletal muscle wasting with recombinant human growth hormone secreted from a tissue-engineered bioartificial muscle

Skeletal muscle wasting is a significant problem in elderly and debilitated patients. Growth hormone (GH) is an anabolic growth factor for skeletal muscle but is difficult to deliver in a therapeutic manner by injection owing to its in vivo instability. A novel method is presented for the sustained secretion of recombinant human GH (rhGH) from genetically modified skeletal muscle implants, which reduces host muscle wasting. Proliferating murine C2C12 skeletal myoblasts stably transduced with the rhGH gene were tissue engineered in vitro into bioartificial muscles (C2-BAMs) containing organized postmitotic myofibers secreting 3-5 microg of rhGH/day in vitro. When implanted subcutaneously into syngeneic mice, C2-BAMs delivered a sustained physiologic dose of 2.5 to 11.3 ng of rhGH per milliliter of serum. rhGH synthesized and secreted by the myofibers was in the 22-kDa monomeric form and was biologically active, based on downregulation of a GH-sensitive protein synthesized in the liver. Skeletal muscle disuse atrophy was induced in mice by hindlimb unloading, causing the fast plantaris and slow soleus muscles to atrophy by 21 to 35% ( < 0.02). This atrophy was significantly attenuated 41 to 55% (p < 0.02) in animals that received C2-BAM implants, but not in animals receiving daily injections of purified rhGH (1 mg/kg/day). These data support the concept that delivery of rhGH from BAMs may be efficacious in treating muscle-wasting disorders.

NASA Discipline Musculoskeletal↗

NASA NTRS1990

Computer aided mechanogenesis of skeletal muscle organs from single cells in vitro

Complex mechanical forces generated in the growing embryo play an important role in organogenesis. Computerized application of similar forces to differentiating skeletal muscle myoblasts in vitro generate three dimensional artificial muscle organs. These organs contain parallel networks of long unbranched myofibers organized into fascicle-like structures. Tendon development is initiated and the muscles are capable of performing directed, functional work. Kinetically engineered organs provide a new method for studying the growth and development of normal and diseased skeletal muscle.

Vandenburgh, Herman H.↗

NASA NTRS1991

Computer-aided mechanogenesis of skeletal muscle organs from single cells in vitro

Vanderburgh, Herman H.↗

NASA NTRS2005

Threshold Gravity Determination and Artificial Gravity Studies Using Magnetic Levitation

What is the threshold gravity (minimum gravity level) required for the nominal functioning of the human system? What dosage is required (magnitude and duration)? Do human cell lines behave differently in microgravity in response to an external stimulus? The critical need for a variable gravity simulator is emphasized by recent experiments on human epithelial cells and lymphocytes on the Space Shuttle clearly showing that cell growth and function are markedly different from those observed terrestrially. Those differences are also dramatic between cells grown in space and those in Rotating Wall Vessels (RWV), or NASA bioreactor often used to simulate microgravity, indicating that although morphological growth patterns (three dimensional growth) can be successfully simulated using RWVs, cell function performance is not reproduced - a critical difference. If cell function is dramatically affected by gravity off-loading, then cell response to stimuli such as radiation, stress, etc. can be very different from terrestrial cell lines. Yet, we have no good gravity simulator for use in study of these phenomena. This represents a profound shortcoming for countermeasures research. We postulate that we can use magnetic levitation of cells and tissue, through the use of strong magnetic fields and field gradients, as a terrestrial microgravity model to study human cells. Specific objectives of the research are: 1. To develop a tried, tested and benchmarked terrestrial microgravity model for cell culture studies; 2. Gravity threshold determination; 3. Dosage (magnitude and duration) of g-level required for nominal functioning of cells; 4. Comparisons of magnetic levitation model to other models such as RWV, hind limb suspension, etc. and 5. Cellular response to reduced gravity levels of Moon and Mars.

Ramachandran, N.↗

NASA NTRS1983

Finite area method for nonlinear supersonic conical flows

A fully conservative numerical method for the computation of steady inviscid supersonic flow about general conical bodies at incidence is described. The procedure utilizes the potential approximation and implements a body conforming mesh generator. The conical potential is assumed to have its best linear variation inside each mesh cell; a secondary interlocking cell system is used to establish the flux balance required to conserve mass. In the supersonic regions the scheme is symmetrized by adding artificial viscosity in conservation form. The algorithm is nearly an order of a magnitude faster than present Euler methods and predicts known results accurately and qualitative features such as nodal point lift off correctly. Results are compared with those of other investigators.

Sritharan, S. S.↗

NASA NTRS1989

The reusable reentry satellite: A new capability for NASA - A vehicle for international cooperation

NASA's LifeSat program, which is designed to study biological systems, is described. The program is also designed to understand how living organisms respond to microgravity as low as 0.00001 G, various levels of artificial gravity up to 1.5 G, and cosmic radiation. Modules to be developed for LifeSat missions include specialized modules to support animals, plants, cells, and tissues.

Ballard, Rodney W.↗

NASA NTRS1972

Pattern of filling in the pulmonary capillary bed.

Artificially ventilated and perfused dog lungs were rapidly frozen under various physiological conditions. In 2-micron-thick sections the number of red blood cells (RBCs) per 10-micron length of alveolar septum was counted. Under conditions in which alveolar pressure exceeded venous pressure, variation in RBC concentration within areas supplied by single arterioles accounted completely for variation between areas supplied by different arterioles. Except at very high perfusion pressures when venous pressure exceeded alveolar pressure, there was no significant correlation between RBC concentrations of pairs of adjacent septa.

Warrell, D. A.↗

NASA NTRS1982

Finite area method for nonlinear conical flows

A fully conservative finite area method for the computation of steady inviscid flow about general conical bodies at incidence is described. The procedure utilizes the potential approximation and implements a body conforming mesh generator. The conical potential is assumed to have its best linear variation inside each mesh cell and a secondary interlocking cell system is used to establish the flux balance required to conserve mass. In the supersonic regions the scheme is desymmetrized by adding appropriate artificial viscosity in conservation form. The algorithm is nearly an order of a magnitude faster than present Euler methods and predicts known results accurately and qualitative features such as nodal point lift off correctly. Results are compared with those of other investigations.

Sritharan, S. S.↗

NASA NTRS1986

Parametric effects of CFL number and artificial smoothing on numerical solutions using implicit approximate factorization algorithm

An implicit approximate factorization algorithm is employed to quantify the parametric effects of Courant number and artificial smoothing on numerical solutions of the unsteady 3-D Euler equations for a windmilling propeller (low speed) flow field. The results show that propeller global or performance chracteristics vary strongly with Courant number and artificial dissipation parameters, though the variation is such less severe at high Courant numbers. Candidate sets of Courant number and dissipation parameters could result in parameter-dependent solutions. Parameter-independent numerical solutions can be obtained if low values of the dissipation parameter-time step ratio are used in the computations. Furthermore, it is realized that too much artificial damping can degrade numerical stability. Finally, it is demonstrated that highly resolved meshes may, in some cases, delay convergence, thereby suggesting some optimum cell size for a given flow solution. It is suspected that improper boundary treatment may account for the cell size constraint.

Daso, E. O.↗

NASA NTRS1994

Numerical solution of the two-dimensional time-dependent incompressible Euler equations

A numerical method is presented for solving the artificial compressibility form of the 2D time-dependent incompressible Euler equations. The approach is based on using an approximate Riemann solver for the cell face numerical flux of a finite volume discretization. Characteristic variable boundary conditions are developed and presented for all boundaries and in-flow out-flow situations. The system of algebraic equations is solved using the discretized Newton-relaxation (DNR) implicit method. Numerical results are presented for both steady and unsteady flow.

Whitfield, David L.↗

NASA NTRS1994

The response to gravity is correlated with the number of statoliths in Chara rhizoids

In contrast to higher plants, Chara rhizoids have single membrane-bound compartments that appear to function as statoliths. Rhizoids were generated by germinating zygotes of Chara in either soil water (SW) medium or artificial pond water (APW) medium. Differential-interference-contrast microscopy demonstrated that rhizoids form SW-grown plants typically contain 50 to 60 statoliths per cell, whereas rhizoids from APW-grown plants contain 5 to 10 statoliths per cell. Rhizoids from SW are more responsive to gravity than rhizoids from APW because (a) SW rhizoids were oriented to gravity during vertical growth, whereas APW rhizoids were relatively disoriented, and (b) curvature of SW rhizoids was 3 to 4 times greater throughout the time course of curvature. The growth rate of APW rhizoids was significantly greater than that of SW-grown rhizoids. This latter result suggests that APW rhizoids are not limited in their ability for gravitropic curvature by growth and that these rhizoids are impaired in the early stages of gravitropism (i.e. gravity perception). Plants grown in APW appeared to be healthy because of their growth rate and the vigorous cytoplasmic streaming observed in the rhizoids. This study is comparable to earlier studies of gravitropism in starch-deficient mutants of higher plants and provides support for the role of statoliths in gravity perception.

Non-NASA Center↗

NASA NTRS2019

Evaluating Precipitation Features and Rainfall Characteristics in a Multi-scale Modeling Framework

Cloud and precipitation systems over the tropics and subtropics are simulated with a multi-scale modeling framework (MMF) and compared against the TRMM radar precipitation features (RPFs) product. A methodology, in close analogy to the TRMM RPFs, is developed to analyze simulated cloud precipitating structures from the embedded two-dimensional cloud-resolving models (CRMs) within an MMF. Despite the two-dimensionality of the CRMs, the simulated RPFs population distribution, and horizontal and vertical structure are in good agreement with TRMM observations. However, some deficits are also found in the model simulations. The model tends to overestimate mean convective precipitation rates for RPFs with a size less than 100 km, contributing to the excessive precipitation biases in the warm pool and western Pacific, western and northern India Ocean, and eastern Pacific commonly found in most MMFs. For large features with a size greater than 150 km, both convective and stratiform rain rates are underestimated. The distribution of maximum radar echo top heights as a function of RPF size is well simulated except the model tends to underestimate the occurrence frequency of maximum heights greater than 15 km. The maximum echo top heights for convective cells embedded within large RPFs with a size greater than 150 km are also underestimated. The cyclic lateral boundary with a limited model domain generates artificial occurrences for RPFs with a size close to the model domain size, producing a significant contribution to the total rainfall due to their sizes. This cyclic lateral boundary effect can be easily identified and quantified in both probability and cumulative distribution functions of RPFs. The geophysical distribution of the population of the largest RPFs in the control experiment shows they are mainly located in the Subtropics but also partially contribute to the common MMF biases of excessive precipitation in the Tropics. Sensitivity experiments using CRMs with different domain sizes and different grid spacings show larger domains (higher resolution) tend to shift the RPFs distribution to large (small) sizes. The cyclic lateral boundary biases increase as CRM domain size decreases. The impacts of model horizontal and vertical resolution on simulated convective systems are also investigated.

Chern, Jiun-Dar↗

NASA NTRS1981

A development of grid generation procedure for multicomponent aerodynamic configuration

Two approaches for solving the transonic flow in a multi-block grid were explored. The first approach examines a method involving "zonal decomposition" wherein block boundaries are treated as true boundary surfaces separating interfacing grids. The issues investigated involve techniques for matching solutions at a block boundary. A feasibility study was completed and the results are presented. The second approach involves overlapping grids for differencing across a block boundary near an artificially induced coordinate singularity occurring at a fictitious corner. This approach selects a set of neighboring nodes for the fictitious corner such that the resulting physical cells for a node are topologically the same as any other node on the airfoil surface.

Chen, H. C.↗

NASA NTRS1991

Entropy-Based Approach To Nonlinear Stability

NASA technical memorandum suggests schemes for numerical solution of differential equations of flow made more accurate and robust by invoking second law of thermodynamics. Proposes instead of using artificial viscosity to suppress such unphysical solutions as spurious numerical oscillations and nonlinear instabilities, one should formulate equations so that rate of production of entropy within each cell of computational grid be nonnegative, as required by second law.

Merriam, Marshal L.↗

NASA NTRS2004

New Gas Polarographic Hydrogen Sensor

Polarography is the measurement of the current that flows in solution as a function of an applied voltage. The actual form of the observed polarographic current depends upon the manner in which the voltage is applied and on the characteristics of the working electrode. The new gas polarographic H2 sensor shows a current level increment with concentration of the gaseous H2 similar to those relating to metal ions in liquid electrolytes in well-known polarography. This phenomenon is caused by the fact that the diffusion of the gaseous H2 through a gas diffusion hole built in the sensor is a rate-determining step in the gaseous-hydrogen sensing mechanism. The diffusion hole artificially limits the diffusion of the gaseous H2 toward the electrode located at the sensor cavity. This gas polarographic H2 sensor. is actually an electrochemical-pumping cell since the gaseous H2 is in fact pumped via the electrochemical driving force generated between the electrodes. Gaseous H2 enters the diffusion hole and reaches the first electrode (anode) located in the sensor cavity to be transformed into an H+ ions or protons; H+ ions pass through the electrolyte and reach the second electrode (cathode) to be reformed to gaseous H2. Gas polarographic 02 sensors are commercially available; a gas polarographic 02 sensor was used to prove the feasibility of building a new gas polarographic H2 sensor.

Dominguez, Jesus A.↗

NASA NTRS2006

H2 Detection via Polarography

Polarography is the measurement of the current that flows in solution as a function of an applied voltage. The actual form of the observed polarographic current depends upon the manner in which the voltage is applied and on the characteristics of the working electrode. The new gas polarographic H2 sensor shows a current level increment with concentration of the gaseous H2 similar to those relating to metal ions in liquid electrolytes in well-known polarography. This phenomenon is caused by the fact that the diffusion of the gaseous H2 through a gas diffusion hole built in the sensor is a rate-determining step in the gaseous-hydrogen sensing mechanism. The diffusion hole artificially limits the diffusion of the gaseous H2 toward the electrode located at the sensor cavity. This gas polarographic H2 sensor is actually an electrochemical-pumping cell since the gaseous H2 is in fact pumped via the electrochemical driving force generated between the electrodes. Gaseous H2 enters the diffusion hole and reaches the first electrode (anode) located in the sensor cavity to be transformed into an H ions or protons; H ions pass through the electrolyte and reach the second electrode (cathode) to be reformed to gaseous H2. Gas polarographic O2 sensors are commercially available; a gas polarographic O2 sensor was used to prove the feasibility of building a new gas polarographic H2 sensor.

Dominquez, Jesus↗

NASA NTRS1975

L-phase variants of Agromyces ramosus

Earlier results suggested that Agromyces ramosus possibly might exist naturally in soil as a cell-wall-defective form. The purpose of the present study was to test this hypothesis by determining whether the laboratory-adapted strains of A. ramosus could be artificially induced into the L-phase and, if so, to examine some parameters affecting induction and the stability of the L-forms. The hypothesis was also tested by attempting to revert the laboratory L-phase strains by subjecting them to the technique originally used for isolation of the bacterial form from soil. It is shown that A. ramosus is easily induced into the L-phase by growing it on an agar media containing low levels of penicillin or glycine. The L-forms are found to be stable after initial contact with the inducing agent and to be unable to be reverted to the bacterial form. However, this lack of reversion does not completely negate the hypothesis that L-forms might occur in nature, because it is possible that L-forms existing in the natural state are less stable than those found in the laboratory where there is little selective pressure toward reversion.

Horwitz, A. H.↗