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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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At least 109 records · Page 6

Hypergravity Stimulates the Extracellular Matrix/Integrin-Signaling Axis and Proliferation in Primary Osteoblasts

We set out to determine the molecular mechanisms involved in the proliferative response of primary rat osteoblasts to mechanical stimulation using cell culture centrifugation as a model for hypergravity. We hypothesized that this proliferative response is mediated by specific integrin/Extracellular Matrix (ECM) interactions. To investigate this question we developed a cell culture centrifuge and an automated system that performs cell fixation during hypergravity loading. We generated expression vectors for various focal adhesion and cytoskeletal proteins fused to GFP or dsRed and visualized these structures in transfected (or infected) osteoblasts. The actin cytoskeleton was also visualized using rhodamine-phalloidin staining and Focal Adhesion Kinase (FAK) levels were assessed biochemically. We observed that a 24 hour exposure to 50-g stimulated proliferation compared to the 1-g control when cells were plated on fibronectin, collagen Type I , and collagen Type IV, but not on uncoated tissue culture plastic surfaces. This proliferative response was greatest for osteoblasts grown on fibronectin (2-fold increase over 1-g control) and collagen Type I (1.4 fold increase over 1-g control), suggesting that specific matrices and integrins are involved in the signaling pathways required for proliferation. Exposing osteoblasts grown on different matrices to 10-g or 25-g showed that effects on proliferation depended on both matrix type and loading level. We found that osteoblasts exposed to a short pulse of hypergravity during adhesion spread further and had more GFP-FAK containing focal adhesions compared to their 1-g controls. While overall levels of FAK did not change, more FAK was in the active (phosphorylated) form under hypergravity than in the 1-g controls. Cytoskeletal F-actin organization into filaments was also more prominent after brief exposures to hypergravity during the first five minutes of adhesion. These results suggest that specific integrins sense hypergravity and activate distinct matrix-dependent FAK signaling pathways that can enhance proliferation. Our results also imply that brief exposures to hypergravity accelerate cell adhesion and spreading processes via the focal adhesion-signaling axis. These results support the role of the ECM/integrin-signaling axis in osteoblast response to hypergravity loading.

Parra, M.↗

Cultural systems for growing potatoes in space

Higher plants are being evaluated for life support to provide needed food, oxygen and water as well as removal of carbon dioxide from the atmosphere. The successful utilization of plants in space will require the development of not only highly productive growing systems but also highly efficient bioregenerative systems. It will be necessary to recycle all inedible plant parts and all human wastes so that the entire complement of elemental compounds can be reused. Potatoes have been proposed as one of the desirable crops because they are 1) extremely productive, yielding more than 100 metric tons per hectare from field plantings, 2) the edible tubers are high in digestible starch (70%) and protein (10%) on a dry weight basis, 3) up to 80% of the total plant production is in tubers and thus edible, 4) the plants are easily propagated either from tubers or from tissue culture plantlets, 5) the tubers can be utilized with a minimum of processing, and 6) potatoes can be prepared in a variety of different forms for the human diet (Tibbitts et al., 1982). However potatoes have a growth pattern that complicates the development of growing the plants in controlled systems. Tubers are borne on underground stems that are botanically termed 'rhizomes', but in common usage termed 'stolons'. The stolons must be maintained in a dark, moist area with sufficient provision for enlargement of tubers. Stems rapidly terminate in flowers forcing extensive branching and spreading of plants so that individual plants will cover 0.2 m2 or more area. Thus the growing system must be developed to provide an area that is darkened for tuber and root growth and of sufficient size for plant spread. A system developed for growing potatoes, or any plants, in space will have certain requirements that must be met to make them a useful part of a life support system. The system must 1) be constructed of materials, and involve media, that can be reused for many successive cycles of plant growth, 2) involve a minimum quantity of media, 3) contain media that is essentially inert and not oxidize or degrade with use, 4) utilize a recirculating nutrient solution to permit regulation of pH and nutrient concentrations, and 5) be capable of complete automation of all planting, maintenance and harvesting procedures.

NASA Discipline Number 61-10↗

Size of tuber propagule influences injury of 'Kennebec' potato plants by constant light

Chlorosis and necrotic spotting develop on the foliage of particular cultivars of potato (Solanum tuberosum L.) when grown under constant light. 'Kennebec', a cultivar severely injured by constant light when propagated from tissue-cultured plantlets, also was injured when plants were propagated from small tuber pieces (approximately 1 g). However, plants did not develop injury when propagated from large tuber pieces (approximately 100 g). Plants from large tuber pieces grew more rapidly than plants from small tuber pieces. The role of plant vigor and carbohydrate translocation in controlling injury development is discussed.

NASA Discipline Life Support Systems↗

The Cytoskeleton and Force Response Mechanisms

The long term aim of this project was to define the mechanisms by which cells sense and respond to the physical forces experienced at 1g and missing in microgravity. Identification and characterization of the elements of the cells force response mechanism could provide pathways and molecules to serve as targets for pharmacological intervention to mitigate the pathologic effects of microgravity. Mechanical forces experienced by the organism can be transmitted to cells through molecules that allow cells to bind to the extracellular matrix and through other types of molecules which bind cells to each other. These molecules are coupled in large complexes of proteins to structural elements such as the actin cytoskeleton that give the cell the ability to sense, resist and respond to force. Application of small forces to tissue culture cells causes local elevation of intracellular calcium through stretch activated ion channels, increased tyrosine phosphorylation and a restructuring of the actin cytoskeleton. Using collagen coated iron oxide beads and strong magnets, we can apply different levels of force to cells in culture. We have found that force application causes the cells to polymerize actin at the site of mechanical deformation and unexpectedly, to depolymerize actin across the rest of the cell. Observations of GFP- actin expressing cells demonstrate that actin accumulates at the site of deformation within the first five minutes of force application and is maintained for many tens of minutes after force is removed. Consistent with the reinforcement of the cytoskeletal structures underlying the integrin-bead interaction, force also alters the motion of bound magnetic beads. This effect is seen following the removal of the magnetic field, and is only partially ablated by actin disruption with cytochalsin B. While actin is polymerizing locally at the site of force application, force also stimulates a global reduction in actin filament content within the cells. We have examined the roles of several actin filament disassembly factors in the global reduction of cellular actin filaments. The calcium regulated actin filament severing protein gelsolin is not necessary for the increased actin turnover, as cells derived from gelsolin null and wildtype mice still show a reduction in total actin filament content. Instead, our work suggests that the actin binding protein cofilin may be important for these changes in actin dynamics. Cofilin binds to and enhances the disassembly of actin filaments. Using immunological methods, we observe transient changes in the phosphorylation state of cofilin upon force application that suggests that cofilin may mediate actin filament turnover. Early after force application, cofilin is transiently dephosphorylated, activating its actin disassembly activity. Subsequently, we find a hyper-phosphorylation of cofilin, rendering it inactive. This reduction in cofilin activity may explain the stability of the force induced actin structuttes. In testing this hypothesis, we aimed to generate cells that express the constituitively active kinase (LIM-kinase) that phosphorylates cofilin. lnidial attempts in the cell lines used for the our previous studies proved unsuccessful. While we prepare this work for pubication, we are continuing to study other cell lines and tissue sources to determine whether they show a reduction in F-actin content after force application.

Allen, Philip Goodwin↗

General session 2

Analyzing action of tetracyclines on calcification in vitro, and data on tissue cultures

Source record↗

Lasers in biology and medicine

Laser applications in biology and medicine covering surgery, ophthalmology, microorganisms, viruses, tissue culture, tumor eradication, radiation hazards, etc

Fine, S.↗

Effects of the space flight environment on man's immune system. II - Lymphocyte counts and reactivity.

The present studies were undertaken to assess the effects of the environment of space flights on the cellular division of the human immune system. Peripheral blood absolute lymphocyte counts were determined at various preflight and postflight intervals for the 21 crewmen of Apollo Missions 7-13. Mean lymphocyte numbers tended to exhibit a delayed significant but fluctuating increase shortly after recovery, although a variety of responses was seen in individual astronauts. The in vitro reactivity of lymphocytes, reflected by RNA and DNA synthesis rates by unstimulated and PHA-stimulated lymphocytes tissue-cultured preflight and postflight from the same participants, was found to remain within previously established normal ranges. These results indicate that functional integrity of cellular immune potential as reflected by in vitro techniques is maintained during this spaceflight experience.

Fischer, G. L.↗

Effects of geophysical extra-terrestrial and terrestrial physical stimuli on living organisms - Effects of gravity fields on living organisms

The biologic effects of greatly reduced gravity resulting from space flight are examined. Aspects of U.S. space biology during the period from 1960 to 1972 are discussed, giving attention to the Discoverer satellites, the Gemini series, the OV1-4 satellite, the biosatellite project, the orbiting frog otolith experiment, and the Apollo program. Other studies considered are related to the effects of galactic particles on nonproliferating cells, a recoverable tissue culture experiment, cell cycle maintenance in human lung cells, and effects of space flight on circadian rhythms. Viking will land on the planet Mars in 1975 in search for life forms.

Saunders, R. J. F.↗

Survey of cell biology experiments in reduced gravity

The effects of spaceflight on terrestrial cell systems are discussed. With some important exceptions, static cell systems carried aboard U.S.A. and U.S.S.R. space flights have failed to reveal space related anomalies. Some sophisticated devices which were developed for viewing directly, or continuously recording, the growth of cells, tissue cultures and eggs in flight, are described and the results summarized. The unique presence of high energy, multicharged (HZE) particles and full-range ultraviolet irradiation in space prompted evaluation of the response of single cells to these factors. Summary results and general conclusions are presented. Potential areas of research in future space flights are identified.

Taylor, G. R.↗

Assessment of pancreas cells

Pancreatic islets were obtained from guinea pig pancreas by the collagenase method and kept alive in tissue culture prior to further studies. Pancreas cell morphology was studied by standard histochemical techniques using light microscopy. Preparative vertical electrophoresis-levitation of dispersed fetal guinea pig pancreas cells was conducted in phosphate buffer containing a heavy water (D20) gradient which does not cause clumping of cells or alter the osmolarity of the buffers. The faster migrating fractions tended to be enriched in beta-cell content. Alpha and delta cells were found to some degree in most fractions. A histogram showing the cell count distribution is included.

Vanoss, C. J.↗

Chemical Growth Regulators for Guayule Plants

Test Tubes containing Guayule - tissue cultures were used in experiments to test effects of chemical-growth regulators. The shoots grew in response to addition of 2-(3,4-dichlorophenoxy)-triethylamine (triethylamine (TEA) derivative) to agar medium. Preliminary results indicate that a class of compounds that promotes growth in soil may also promote growth in a culture medium. Further experiments are needed to define the effect of the TEA derivative.

Dastoor, M. N.↗

US plant and radiation dosimetry experiments flown on the Soviet satellite Cosmos 1129

Experiments included: 30 young male Wistar SPF rats used for wide range physiological studies; experiments with plants, fungi, insects, and mammalian tissue cultures; radiation physics experiments; a heat convection study; a rat embryology experiment in which an attempt was made to breed 2 male and 5 female rats during the flight; and fertile quail eggs used to determine the effects of spaceflight on avian embryogenesis. Specimens for US experiments were initially prepared at the recovery site or in Moscow and transferred to US laboratories for complete analyses. An overview of the mission focusing on preflight, on orbit, and postflight activities pertinent to the fourteen US experiments aboard Cosmos 1129 is presented.

Heinrich, M. R.↗

Development of guayule (parthenium argentatum)

The cytokinin benzylaminopurine strongly stimulated shoot growth, and the number of regenerated buds on the inoculum was proportional to its concentration. These buds produced shoots several centimeters in length which were caused to root on medium containing indolebutyric acid. Transferred to the septic condition of soil, the plantlets were gradually brought into full sunlight where they showed a brief vegetative growth with production of mature type leaves, and flowered. In contrast, seedlings of the same age remained vegetative. Chromosome studies of root smears from the tissue cultured plantlets showed that 2n = 36, the normal number for sexually reproducing guayules.

Ball, E. A.↗

The Antaeus Project - An orbital quarantine facility for analysis of planetary return samples

A design is presented for an earth-orbiting facility for the analysis of planetary return samples under conditions of maximum protection against contamination but minimal damage to the sample. The design is keyed to a Mars sample return mission profile, returning 1 kg of documented subsamples, to be analyzed in low earth orbit by a small crew aided by automated procedures, tissue culture and microassay. The facility itself would consist of Spacelab shells, formed into five modules of different sizes with purposes of power supply, habitation, supplies and waste storage, the linking of the facility, and both quarantine and investigation of the samples. Three barriers are envisioned to protect the biosphere from any putative extraterrestrial organisms: sealed biological containment cabinets within the Laboratory Module, the Laboratory Module itself, and the conditions of space surrounding the facility.

Sweet, H. C.↗

Space Station and the life sciences

Previous fundamental research in space life sciences is examined, and consideration is devoted to studies relevant to Space Station activities. Microgravity causes weight loss, hemoconcentration, and orthostatic intolerance when astronauts returns to earth. Losses in bone density, bone calcium, and muscle nitrogen have also been observed, together with cardiovascular deconditioning, fluid-electrolyte metabolism alteration, and space sickness. Experiments have been performed with plants, bacteria, fungi, protozoa, tissue cultures, invertebrate species, and with nonhuman vertebrates, showing little effect on simple cell functions. The Spacelab first flight will feature seven life science experiments and the second flight, two. Further studies will be performed on later flights. Continued life science studies to optimize human performance in space are necessary for the efficient operation of a Space Station and the assembly of large space structures, particularly in interaction with automated machinery.

White, R. J.↗