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Todd, Paul

Publications and source records attributed to Todd, Paul.

At least 19 records

Multistage Electrophoretic Separators

A multistage electrophoresis apparatus has been invented for use in the separation of cells, protein molecules, and other particles and solutes in concentrated aqueous solutions and suspensions. The design exploits free electrophoresis but overcomes the deficiencies of prior free-electrophoretic separators by incorporating a combination of published advances in mathematical modeling of convection, sedimentation, electro-osmotic flow, and the sedimentation and aggregation of droplets. In comparison with other electrophoretic separators, these apparatuses are easier to use and are better suited to separation in relatively large quantities characterized in the art as preparative (in contradistinction to smaller quantities characterized in the art as analytical). In a multistage electrophoretic separator according to the invention, an applied vertical steady electric field draws the electrically charged particles of interest from within a cuvette to within a collection cavity that has been moved into position of the cuvette. There are multiple collection cavities arranged in a circle; each is aligned with the cuvette for a prescribed short time. The multistage, short-migration-path character of the invention solves, possibly for the first time, the fluid-instability problems associated with free electrophoresis. The figure shows a prototype multistage electrophoretic separator that includes four sample stations and five collection stages per sample. At each sample station, an aqueous solution or suspension containing charged species to be separated is loaded into a cuvette, which is machined into a top plate. The apparatus includes a lower plate, into which 20 collection cavities have been milled. Each cavity is filled with an electrophoresis buffer solution. For the collection of an electrophoretic fraction, the lower plate is rotated to move a designated collection cavity into alignment with the opening of the cuvette. An electric field is then applied between a non-gassing electrode in the collection cavity and an electrolyte compartment, which is separated from the cuvette by a semipermeable membrane. The electrolyte is refreshed by circulation by use of a peristaltic pump. In subsequent steps, the lower plate is rotated to collect other electrophoretic fractions. Later, the collected fractions are removed from the collection cavities through ports that have threaded plugs. The base of the apparatus contains power supplies and a computer interface. The design includes provisions for monitoring and feedback control of cavity position, electric field, and temperature. The operation of the apparatus can easily be automated, as demonstrated by use of software that has already been written for this purpose.

Thomas, Nathan

Diet as a factor in behavioral radiation protection following exposure to heavy particles

Major risks associated with radiation exposures on deep space missions include carcinogenesis due to heavy-particle exposure of cancer-prone tissues and performance decrements due to neurological damage produced by heavy particles. Because exposure to heavy particles can cause oxidative stress, it is possible that antioxidants can be used to mitigate these risks (and possibly some health risks of microgravity). To assess the capacity of antioxidant diets to mitigate the effects of exposure to heavy particles, rats were maintained on antioxidant diets containing 2% blueberry or strawberry extract or a control diet for 8 weeks prior to exposure to 1.5 or 2.0 Gy of accelerated iron particles at Brookhaven National Laboratory. Following irradiation rats were tested on a series of behavioral tasks: amphetamine-induced taste aversion learning, operant responding and spatial learning and memory. The results indicated that the performance of the irradiated rats maintained on the antioxidant diets was, in general, significantly better than that of the control animals, although the effectiveness of the diets ameliorating the radiation-induced deterioration in performance varied as a function of both the specific diet and the specific endpoint. In addition, animals fed antioxidant diets prior to exposure showed reduced heavy particle-induced tumorigenesis one year after exposure compared to the animals fed the control diet. These results suggest that antioxidant diets have the potential to serve as part of a system designed to provide protection to astronauts against the effects of heavy particles on exploratory missions outside the magnetic field of the earth.

Non-NASA Center

Experiments with osteoblasts cultured under hypergravity conditions

To understand further the role of gravity in osteoblast attachment, osteoblasts were subjected to hypergravity conditions in vitro. Scanning electron microscopy of all confluent coverslips from FPA units show that the number of attached osteoblasts was similar among gravitational levels and growth durations (~90 cells/microscopic field). Specifically, confluent 1.0 G control cultures contained an average of 91 +/- 8 cells/field, 3.3 G samples had 88 +/- 8 cells/field, and 4.0 G cultures averaged 90 +/- 7 cells/field. The sparsely plated cultures assessed by immunohistochemistry also had similar numbers of cells at each time point (l.0 G was similar to 3.3 and 4.0 G), but cell number changed from one time point to the next as those cells proliferated. Immunohistochemistry of centrifuged samples showed an increase in number (up to 160% increase) and thickness (up to 49% increase) of actin fibers, a decrease in intensity of fibronectin fluorescence (18-23% decrease) and an increase in number of vinculin bulbs (202-374% increase in number of vinculin bulbs/area). While hypergravity exposure did not alter the number of attached osteoblasts, it did result in altered actin, fibronectin, and vinculin elements, changing some aspects of osteoblast- substrate adhesion.

NASA Program Fundamental Space Biology

Biological countermeasures in space radiation health

Exposure to the types of ionizing radiation encountered during space travel may cause a number of health-related problems, but the primary concern is related to the increased risk of cancer induction in astronauts. The major types of radiation considered to be of importance during space travel are protons and particles of high atomic number and high energy (HZE particles). It is now clear that biological countermeasures can be used to prevent or reduce the levels of biological consequences resulting from exposure to protons or HZE particles, including the induction of cancer, immunosuppression and neurological defects caused by these types of ionizing radiation. Research related to the dietary additions of agents to minimize the risks of developing health-related problems which can result from exposure to space radiations is reviewed.

NASA Discipline Radiation Health

Influence of Solutocapillary Convection on Macrovoid Defect Formation in Polymeric Membranes

The focus of this research project involved the dry-cast process for polymeric membrane formation, whereby evaporation of solvent from an initially homogeneous polymer/solvent/ nonsolvent solution results in phase separation and the formation of polymer-rich and polymer-lean phases. Under certain conditions the polymer-lean phase gives rise to very large and usually undesirable, tear-drop-shaped pores (size approx. 10 - 50 microns) termed macrovoids (MVs). Although in many cases the presence of MV pores has deleterious effects on membrane performance, there are a number of innovative applications where the presence of such pores is highly desirable. Although researchers have proposed a variety of mechanisms for MV formation over the past three decades, two main hypotheses are currently favored: one asserts that MV growth can be attributed solely to diffusion (the diffusive growth hypothesis), whereas the other states that solutocapillary convection (the SC hypothesis) at the MV interface contributes to growth. The overall goal of this research was to obtain a more comprehensive understanding of the fundamental mechanism of MV growth. This research incorporates a coupled modeling and experimental approach to test a solutocapillary convection hypothesis for the growth of macrovoid pores in polymeric membranes. Specifically, we utilized a modification of the first principles model developed by two of the PIs (ARG and WBK) for dry-cast CA membranes. For the experimental component, two separate and mutually complementary approaches were used to study MV growth. In the first, membranes cast in a zero-g environment aboard the NASA KC-135 aircraft were compared with those cast on the ground to assess the effect of the buoyancy force on membrane morphology and MV size and shape. In the second approach, videomicroscopy flow visualization (VMFV) was utilized to observe MV formation and growth in real time and to assess the effect of surface tension on the MV growth dynamics. As a result of these fundamental studies, our research group advanced a new hypothesis for MV pore development in polymeric membranes.

Greenberg, Alan R.

Investigating the Effect of Impurities on Macromolecule Crystal Growth in Microgravity

Chicken egg-white lysozyme (CEWL) crystals were grown in microgravity and on the ground in the presence of various amounts of a naturally occurring lysozyme dimer impurity. No significant favorable differences in impurity incorporation between microgravity and ground crystal samples were observed. At low impurity concentration the microgravity crystals preferentially incorporated the dimer. The presence of the dimer in the crystallization solutions in microgravity reduced crystal size, increased mosaicity and reduced the signal to noise ratio of the X-ray data. Microgravity samples proved more sensitive to impurity. Accurate indexing of the reflections proved critical to the X-ray analysis. The largest crystals with the best X-ray diffraction properties were grown from pure solution in microgravity.

Snell, Edward H.

Preparation and Analysis of RNA Crystals

The crystallization of RiboNucleic Acids (RNA) was studied from the standpoint of mechanisms of crystal growth in three tasks: (1) preparation of high-quality crystals of oligonuclotides for X-ray diffraction, (2) finding pathways to the growth of high-quality crystals for X-ray diffraction and (3) investigation of mechanisms of action of inertial acceleration on crystal growth. In these tasks: (1) RNA crystals were prepared and studied by X-ray diffraction; (2) a pathway to high-quality crystals was discovered and characterized; a combination of kinetic and equilibrium factors could be optimized as described below; and (3) an interplay between purity and gravity was found in a combination of space and ground experiments with nucleic acids and proteins. Most significantly, the rate of concentration of precipitant and RNA can be controlled by membrane-based methods of water removal or by diffusion of multivalent cations across an interface stabilized by a membrane. Oligonucleotide solutions are electrokinetically stabilized colloids, and crystals can form by the controlled addition of multivalent cations.

Todd, Paul

Macromolecule Crystal Quality Improvement in Microgravity: The Role of Impurities

While macromolecule impurities may affect crystal size and morphology the over-riding question is; "How do macromolecule impurities effect crystal X-ray quality and diffraction resolution?" In the case of chicken egg white lysozyme, crystals can be grown in the presence of a number of impurities without affecting diffraction resolution. One impurity however, the lysozyme dimer, does negatively impact the X-ray crystal properties. Crystal quality improvement as a result of better partitioning of this impurity during crystallization in microgravity has been reported'. In our recent experimental work dimer partitioning was found to be not significantly different between the two environments. Mosaicity analysis of pure crystals showed a reduced mosaicity and increased signal to noise for the microgravity grown crystals. Dimer incorporation however, did greatly reduce the resolution limit in both ground and microgravity grown crystals. These results indicate that impurity effects in microgravity are complex and may rely on the conditions or techniques employed.

Judge, Russell A.

The Question of Impurities in Macromolecule Crystal Quality Improvement in Microgravity

While macromolecule impurities may affect crystal size and morphology the over-riding question is how do macromolecule impurities effect crystal X-ray quality and diffraction resolution. In the case of chicken egg white lysozyme previous researchers have reported that crystals grown in the presence of ovalbumin, ovotransferrin, and turkey egg white lysozyme show no difference in diffraction resolution compared to those grown in pure solutions. One impurity however, a naturally occurring lysozyme dimer, does negatively impact the X-ray crystal properties. For this impurity it has been reported that crystal quality improvement in microgravity may be due to improved impurity partitioning during crystallization. In this study we have examined the incorporation of the dimer into lysozyme crystals, both on the ground and in microgravity experiments, and have performed detailed X-ray analysis of the crystals using a new technique for finely probing the mosaicity of the crystal at the Stanford Synchrotron Radiation Laboratory. Dimer partitioning was not significantly different in microgravity compared to the ground based experiments, although it is significantly better than that previously reported in microgravity. Mosaicity analysis of pure crystals, 1422 indexed reflections (microgravity) and 752 indexed reflections (ground), gave average results of 0.0066 and 0.0092 degrees (FWHM) respectively. The microgravity crystals also provided an increased signal to noise. Dimer incorporation increased the average mosaicity in microgravity but not on the ground. However, dimer incorporation did greatly reduce the resolution limit in both ground and microgravity grown crystals. The data is being treated anisotropically to explore these effects. These results indicate that impurity effects in microgravity are complex and that the conditions or techniques employed may greatly affect the role of impurities.

Judge, Russell A.

Effect of Pigmentation in Particulate Formation from Fluoropolymer Thermodegradation in Microgravity

Fires aboard spacecraft have occurred as a result of overheated electrical wires and thermodegradation of their insulation, which is composed of fluoropolymers. The particulate products of polymer thermodegradation are only 20-50 run in diameter and are thought to play a role in "polymer fume fever". Therefore an experimental study of the particulates produced by intense ohmic heating of various fluoropolymer-insulated 20 AWG copper wire (representative of spacecraft materials) was undertaken in normal gravity and in microgravity. The 2.2 s drop facility at NASA LeRC and 1.5 s drop facility at the Colorado School of Mines were used to achieve low gravity conditions. Thermophoretic sampling was used for particulate collection. Transmission electron microscopy (TEM) and scanning transmission electron microscopy (STEM) were used to characterize the smoke particulates. It was found that the color of PTFE (Polytetrafluoroethylene) insulation has an overwhelming effect on the size, shape, morphology and, composition of the particulates. Size distributions and shape analyses using computerized image analysis showed that particle size distributions were also dependent on the pigment of the fluoropolymer insulation. The influence of pigment was observed in experiments under both normal and microgravity. Under microgravity conditions, owing to the lack of natural convective transport of particulates, much more particle aggregation was observed, and the nature of the aggregates was dependent on the color of the insulation.

Srivastava, Rajiv

Combustion of PTFE: The Effects of Gravity and Pigmentation on Ultrafine Particle Generation

Ultrafine particles generated during polymer thermodegradation are a major health hazard, owing to their unique pathway of processing in the lung. This hazard in manned spacecraft is poorly understood, because the particulate products of polymer thermodegradation are generated under low gravity conditions. Particulate generated from the degradation of PolyTetraFluoroEthylene (PTFE), insulation coating for 20 AWG copper wire (representative of spacecraft application) under intense ohmic heating were studied in terrestrial gravity and microgravity. Microgravity tests were done in a 1.2-second drop tower at the Colorado School of Mines (CSM). Thermophoretic sampling was used for particulate collection. Transmission Electron Microscopy (TEM) and Scanning Transmission Electron Microscopy (STEM) were used to examine the smoke particulates. Image software was used to calculate particle size distribution. In addition to gravity, the color of PTFE insulation has an overwhelming effect on size, shape and morphology of the particulate. Nanometer-sized primary particles were found in all cases, and aggregation and size distribution was dependent on both color and gravity; higher aggregation occurred in low gravity. Particulates from white, black, red and yellow colored PTFE insulations were studied. Elemental analysis of the particulates shows the presence of inorganic pigments.

McKinnon, J. Thomas

Cell and Particle Interactions and Aggregation During Electrophoretic Motion

The stability and pairwise aggregation rates of small spherical particles under the collective effects of buoyancy-driven motion and electrophoretic migration are analyzed. The particles are assumed to be non-Brownian, with thin double-layers and different zeta potentials. The particle aggregation rates may be enhanced or reduced, respectively, by parallel and antiparallel alignments of the buoyancy-driven and electrophoretic velocities. For antiparallel alignments, with the buoyancy-driven relative velocity exceeding the electrophoretic relative velocity between two widely-separated particles, there is a 'collision-forbidden region' in parameter space due to hydrodynamic interactions; thus, the suspension becomes stable against aggregation.

Wang, Hua

Combustion of PTFE: The effects of gravity on ultrafine particle generation

The objective of this project is to obtain an understanding of the effect of gravity on the toxicity of ultrafine particle and gas phase materials produced when fluorocarbon polymers are thermally degraded or burned. The motivation for the project is to provide a basic technical foundation on which policies for spacecraft health and safety with regard to fire and polymers can be formulated.

McKinnon, Thomas

Electrophoretic interactions and aggregation of colloidal biological particles

The separation of cells or particles from solution has traditionally been accomplished with centrifuges or by sedimentation; however, many particles have specific densities close to unity, making buoyancy-driven motion slow or negligible, but most cells and particles carry surface charges, making them ideal for electrophoretic separation. Both buoyancy-driven and electrophoretic separation may be influenced by hydrodynamic interactions and aggregation of neighboring particles. Aggregation by electrophoresis was analyzed for two non-Brownian particles with different zeta potentials and thin double layers migrating through a viscous fluid. The results indicate that the initial rate of electrophoretically-driven aggregation may exceed that of buoyancy-driven aggregation, even under conditions in which buoyancy-driven relative motion of noninteracting particles is dominant.

Davis, Robert H.

Transport phenomena in the crystallization of lysozyme by osmotic dewatering and liquid-liquid diffusion in low gravity

Two methods of protein crystallization, osmotic dewatering and liquid-liquid diffusion, like the vapor diffusion (hanging-drop and sessile-drop) methods allow a gradual approach to supersaturation conditions. The crystallization of hen egg-white lysozyme, an extensively characterized protein crystal, in the presence of sodium chloride was used as an experimental model with which to compare these two methods in low gravity and in the laboratory. Comparisons of crystal growth rates by the two methods under the two conditions have, to date, indicated that the rate of crystal growth by osmotic dewatering is nearly the same in low gravity and on the ground, while much faster crystal growth rates can be achieved by the liquid-liquid diffusion method in low gravity.

Todd, Paul

Physics, chemistry and pulmonary sequelae of thermodegradation events in long-mission space flight

An event in which electronic insulation consisting of polytetrafluoroethylene undergoes thermodegradation on the Space Station Freedom is considered experimentally and theoretically from the initial chemistry and convective transport through pulmonary deposition in humans. The low-gravity enviroment impacts various stages of event simulation. Vapor-phase and particulate thermodegradation products were considered as potential spacecraft contaminants. A potential pathway for the production of ultrafine particles was identified. Different approaches to the simulation and prediction of contaminant transport were studied and used to predict the distribution of generic vapor-phase products in a Space Station model. A lung transport model was used to assess the pulmonary distribution of inhaled particles, and, finally, the impact of adaptation to low gravity on the human response to this inhalation risk was explored on the basis of known physiological modifications of the immune, endocrine, musculoskeletal and pulmonary systems that accompany space flight.

Todd, Paul

Clinostats and Centrifuges: Their Use, Value, and Limitations in Gravitational Biological Research

The present volume addresses physical phenomena and effects associated with clinostat and centrifuge operations as well as their physiological effects. Particular attention is given to the simulation of the gravity conditions on the ground, the internal dynamics of slowly rotating biological systems, and qualitative and quantitative aspects of the fast-rotating clinostat as a research tool. Also discussed are the development and use of centrifuges in gravitational biology, the use of centrifuges in plant gravitational biology and a comparison of ground-based and flight experiment results, the ability of clinostat to mimic the effect of microgravity on plant cells and organs, and the impact of altered gravity conditions on early EGF-induced signal transduction in human epidermal A431 cells.

Halstead, Thora W.

Physical effects at the cellular level under altered gravity conditions

Several modifications of differentiated functions of animal cells cultivated in vitro have been reported when cultures have been exposed to increased or decreased inertial acceleration fields by centrifugation, clinorotation, and orbital space flight. Variables modified by clinorotation conditions include inertial acceleration, convection, hydrostatic pressure, sedimentation, and shear stress, which also affect transport processes in the extracellular chemical environment. Autocrine, paracrine and endocrine substances, to which cells are responsive via specific receptors, are usually transported in vitro (and possibly in certain embryos) by convection and in vivo by a circulatory system or ciliary action. Increased inertial acceleration increases convective flow, while microgravity nearly abolishes it. In the latter case the extracellular transport of macromolecules is governed by diffusion. By making certain assumptions it is possible to calculate the Peclet number, the ratio of convective transport to diffusive transport. Some, but not all, responses of cells in vitro to modified inertial environments could be manifestations of modified extracellular convective flow.

Todd, Paul