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At least 379 records · Page 21

Non-conventional approaches to food processing in CELSS. I - Algal proteins: Characterization and process optimization

Protein isolate obtained from green algae (Scenedesmus obliquus) cultivated under controlled conditions was characterized. Molecular weight determination of fractionated algal proteins using SDS-polyacrylamide gel electrophoresis revealed a wide spectrum of molecular weights ranging from 15,000 to 220,000. Isoelectric points of dissociated proteins were in the range of 3.95 to 6.20. Amino acid composition of protein isolate compared favorably with FAO standards. High content of essential amino acids leucine, valine, phenylalanine and lysine makes algal protein isolate a high quality component of CELSS diets. To optimize the removal of algal lipids and pigments supercritical carbon dioxide extraction (with and without ethanol as a co-solvent) was used. Addition of ethanol to supercritical CO2 resulted in more efficient removal of algal lipids and produced protein isolate with a good yield and protein recovery. The protein isolate extracted by the above mixture had an improved water solubility.

Nakhost, Z.↗

Comparison of bioseparation methods for microgravity experiments

The efficiency of the 1-g version of the continuous-flow electrophoresis (CFE) system flown on Space Shuttle missions is compared with the efficiency of a commercial CFE for separating living cells (human kidney, liver, and pituitary-gland cells and T-lymphocytes). In addition, the CFE system and a reciprocal isoelectric focusing (RIEF) system are compared with respect to protein pyrification efficiency. Correlations were made among electrophoretic mobilities (EPMs), secretory functions of cells, and input sample concentrations. A significant reduction in mean and range EPM was observed when input sample concentrations exceeded a low threshohold. This effect was not observed in microgravity experiments conducted at sample concentrations three times greater than the threshold for the controls. Comparison of CFE and RIEF methods showed that there are apparent advantages for each method depending on the product. For example, RIEF purification of urokinase removed more protein impurities, but focused the enzyme at a pH different than the enzyme's known isoelectric point.

Morrison, Dennis R.↗

Continuous flow electrophoretic separation of proteins and cells from mammalian tissues

This paper describes an apparatus for continuous flow electrophoresis (CFE), designed to separate macromolecules and cells at conditions of microgravity. In this CFE, buffer flows upward in a 120-cm long flow chamber, which is 16-cm wide x 3.0-mm thick in the microgravity version (and 6-cm wide x 1.5-mm thick in the unit-gravity laboratory version). Ovalbumin and rat serum albumin were separated in space (flight STS-4) with the same resolution of the two proteins achieved at 25 percent total w/v concentration that was obtained in the laboratory at 0.2 percent w/v concentration. Rat anterior pituitary cells, cultured human embryonic kidney cells, and canine Langerhans cells were separated into subpopulations (flight STS-8) more effectively than in unit gravity, with comparable resolution having been achieved at 100 times the concentration possible on earth.

Hymer, W. C.↗

Bioseparations - Flight experiment support research

While important principles of electrokinetic technology, such as electrophoresis, have been demonstrated for microgravity environments and promise to be able to alleviate many of the practical limitations imposed by gravity, the minimal crew time and sample-handling facilities available in orbit call for almost total reliance on postflight analysis of separated fractions. Since cell separations are the most difficult to accomplish in 1-g, two model cell types have been studied intensively to determine the basic relationships between cell surface charge and secretory functions. A rationale has been developed for selecting and qualifying new cell types and proteins that warrant processing in microgravity.

Morrison, Dennis R.↗

Evaluation of available analytical techniques for monitoring the quality of space station potable water

To assure the quality of potable water (PW) on the Space Station (SS) a number of chemical and physical tests must be conducted routinely. After reviewing the requirements for potable water, both direct and indirect analytical methods are evaluated that could make the required tests and improvements compatible with the Space Station operation. A variety of suggestions are made to improve the analytical techniques for SS operation. The most important recommendations are: (1) the silver/silver chloride electrode (SB) method of removing I sub 2/I (-) biocide from the water, since it may interfere with analytical procedures for PW and also its end uses; (2) the orbital reactor (OR) method of carrying out chemistry and electrochemistry in microgravity by using a disk shaped reactor on an orbital table to impart artificial G force to the contents, allowing solution mixing and separation of gases and liquids; and (3) a simple ultra low volume highly sensitive electrochemical/conductivity detector for use with a capillary zone electrophoresis apparatus. It is also recommended, since several different conductivity and resistance measurements are made during the analysis of PW, that the bipolar pulse measuring circuit be used in all these applications for maximum compatibility and redundancy of equipment.

Geer, Richard D.↗

Controlled method of reducing electrophoretic mobility of various substances

A method of reducing electrophoretic mobility of macromolecules, particles, cells, and the like is provided. The method comprises interacting the particles or cells with a polymer-linked affinity compound composed of: a hydrophilic neutral polymer such as polyethylene glycol, and an affinity component consisting of a hydrophobic compound such as a fatty acid ester, an immunocompound such as an antibody or active fragment thereof or simular macromolecule, or other ligands. The reduction of electrophoretic mobility achieved is directly proportional to the concentration of the polymer-linked affinity compound employed, and the mobility reduction obtainable is up to 100 percent for particular particles and cells. The present invention is advantageous in that analytical electrophoretic separation can not be achieved for macromolecules, particles, and cells whose native surface charge structure had prevented them from being separated by normal electrophoretic means. Depending on the affinity component utilized, separation can be achieved on the basis of specific/irreversible, specific/reversible, semi-specific/reversible, relatively nonspecific/reversible, or relatively nonspecific/irreversible ligand-substance interactions. The present method is also advantageous in that it can be used in a variety of standard laboratory electrophoresis equipment.

Vanalstine, James M.↗

Laser light scattering review

Since the development of laser light sources and fast digital electronics for signal processing, the classical discipline of light scattering on liquid systems experienced a strong revival plus an enormous expansion, mainly due to new dynamic light scattering techniques. While a large number of liquid systems can be investigated, ranging from pure liquids to multicomponent microemulsions, this review is largely restricted to applications on Brownian particles, typically in the submicron range. Static light scattering, the careful recording of the angular dependence of scattered light, is a valuable tool for the analysis of particle size and shape, or of their spatial ordering due to mutual interactions. Dynamic techniques, most notably photon correlation spectroscopy, give direct access to particle motion. This may be Brownian motion, which allows the determination of particle size, or some collective motion, e.g., electrophoresis, which yields particle mobility data. Suitable optical systems as well as the necessary data processing schemes are presented in some detail. Special attention is devoted to topics of current interest, like correlation over very large lag time ranges or multiple scattering.

Schaetzel, Klaus↗

Particle migration in suspensions by thermocapillary or electrophoretic motion

Two problems of similar mathematical structure are presented: the thermocapillary motion of bubbles and the electrophoresis of colloidal particles. For both problems, it is shown that in a cloud of n particles surrounded by an infinite expanse of fluid, the velocity of each sphere under creeping flow conditions is equal to the velocity of an isolated particle, unchanged by interactions between the particles. However, when the cloud fills a container, conservation of mass shows that this result cannot continue to hold, and the average translational velocity must be calculated subject to a constraint on the mass flux. It is concluded that the average thermocapillary or electrophoretic translational velocity of a particle in the cloud is related to the effective conductivity of the cloud over the whole range of particle volume fractions, provided that the particles are identical, non-conducting and, for the thermocapillary problem, inviscid.

Acrivos, A.↗

Variable-Speed Instrumented Centrifuges

Report describes conceptual pair of centrifuges, speed of which varied to produce range of artificial gravities in zero-gravity environment. Image and data recording and controlled temperature and gravity provided for 12 experiments. Microprocessor-controlled centrifuges include video cameras to record stop-motion images of experiments. Potential applications include studies of effect of gravity on growth and on production of hormones in corn seedlings, experiments with magnetic flotation to separate cells, and electrophoresis to separate large fragments of deoxyribonucleic acid.

Chapman, David K.↗

Cell separation and electrofusion in space

In microgravity, free-fluid electrophoretic methods for separating living cells and proteins are improved significantly by the absence of gravity-driven phenomena. Cell fusion, culture, and other bioprocessing steps are being investigated to understand the limits of earth-based processing. A multistep space bioprocess is described that includes electrophoretic separation of human target cells, single-cell manipulations using receptor-specific antibodies, electrofusion to produce immortal hybridomas, gentle suspension culture, and monoclonal antibody recovery using continuous-flow electrophoresis or recirculating isoelectric focusing. Improvements in several key steps already have been demonstrated by space experiments, and others will be studied on Space Station Freedom.

Morrison, D. R.↗

Further analyses of human kidney cell populations separated on the Space Shuttle

Cultured human embryonic kidney cells were separated into electrophoretic subpopulations in laboratory experiments and in two separation experiments on the STS-8 (Challenger) Space Shuttle flight using the mid-deck Continuous Flow Electrophoretic Separator (CFES). Populations of cells from each fraction were cultured for the lifetime of the cells, and supernatant medium was withdrawn and replaced at 4-day intervals. Withdrawn medium was frozen at -120 C for subsequent analysis. Enzyme assays, antibodies and gel electrophoresis were used as analytical tools for the detection and quantization of plasminogen activators in these samples. These assays of frozen-culture supernatant fluids confirmed the electrophoretic separation of plasminogen-activator-producing cells from nonproducing cells, the isolation of cells capable of sustained production, and the separation of cells that produce different plasminogen activators from one other.

Stewart, Robin M.↗

Comparison of membrane ATPases from extreme halophiles isolated from ancient salt deposits

Halophilic microorganisms were isolated from Triassic and Permian salt deposits. Two were rods and grew as red colonies; another was a coccus and produced pink colonies. The rods lysed in solutions that lacked added sodium chloride. Growth of all isolates was inhibited by aphidicolin and their bulk-proteins were acidic as judged from isoelectric focusing. Therefore, these organisms were tentatively identified as extreme halophiles. Whole cell proteins patterns of the isolates following gel electrophoresis were distinct and differed from those of representative type strains of halophilic bacteria. The membrane ATPases from the rods were similar to the enzyme from Halobacterium saccharovorum with respect to subunit composition, enzymatic properties and immunological cross-reaction, but differed slightly in amino acid composition. If the age of the microbial isolated is similar to that of the salt deposits, they can be considered repositories of molecular information of great evolutionary interest.

Stan-Lotter, Helga↗

Integrating payload design, planning, and control in the Dutch Utilisation Centre

Spacecraft payload design, experiment planning and scheduling, and payload control are traditionally separate areas of activity. This paper describes the development of a prototype software tool--the Activity Scheduling System (ASS)--which integrates these activity areas. ASS is part of a larger project to build a Dutch Utilisation Centre (DUC), intended eventually to support all space utilization activities in The Netherlands. ASS has been tested on the High Performance Capillary Electrophoresis payload. The paper outlines the integrated preparation and operations concept embodied in ASS. It describes the ASS prototype, including a typical session. The results of testing are summarized. Possible enhancement of ASS, including integration into DUC, is sketched.

Grant, T. J.↗

Role of dielectric constant in electrohydrodynamics of conducting fluids

Electrohydrodynamic sample distortion during continuous flow electrophoresis is an experiment to be conducted during the second International Microgravity Laboratory (IML-2) in July 1994. The specific objective of this experiment is the distortion caused by the difference in dielectric constant between the sample and surrounding buffer. Although the role of sample conductivity in electrohydrodynamic has been the subject of both flight and ground experiments, the separate role of dielectric constant, independent of sample conductivity, has not been measured. This paper describes some of the laboratory research and model development that will support the flight experiment on IML-2.

Rhodes, Percy H.↗

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.↗

Electrokinetic transport of heterogeneous particles in suspension

The Smoluchowski equation for electrophoresis predicts that the electrophoretic velocity of a particle is proportional to its zeta potential but not its size, shape, or orientation. Furthermore, the equation predicts that the rotation rate is identically zero. The Smoluchowski equation fails for heterogeneous particles (i.e., those with nonuniform zeta potentials). Recent theories and experiments show that particles with a dipole moment of zeta potential rotate into alignment with an externally applied electric field. For doublets (particles composed of two spheres) the rotation rate depends on (1) whether the spheres are rigidly rocked or freely rotating, and (2) the gap distance between the spheres. The relative configuration of two coagulated spheres is determined by the colloidal forces of the system. The goal of our research is to use measurements of electrophoretic rotation to determine the gap between two spheres of a colloidal doublet and also to determine whether or not the doublet is rigid.

Velegol, D.↗

Electrophoretic separation of cells and particles from rat pituitary

In spite of the fact that a vast majority of the electrophoresis effort (approximately 90%) could not be done on this mission (IML-2) due to failure of FFEU hardware, we find some interesting differences in flight samples obtained from other parts of the experiment. These differences are entirely novel and sometimes unexpected. This report is organized into 4 parts. Each part describes the data collected thus far from each of the 4 cell culture kits (CCK) which flew in space. Each CCK was loaded with 40x10(exp 6) fresh pituitary cells; all CCK's were identical at the start of the experiment because we prepared one pool of cells.

Hymer, W. C.↗