Rapid recording microspectrophotometer.
Rapid recording microspectrophotometer for IR to UV absorption spectra of living cells, cell organelles and half micron diameter particles
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Rapid recording microspectrophotometer for IR to UV absorption spectra of living cells, cell organelles and half micron diameter particles
The pathogenesis of oxygen toxicity in the lung of rats was studied by electron microscopy. The following long-term effects were established: (1) a progressive destruction of the blood-air barrier beginning with the endothelial cell layer; (2) a profuse edema in the interstitial spaces in the pleural space in the alveoli and in the cytoplasm and organelles; (3) a continuing increase in the quantity and complexity of the alveolar exudate; (4) gradual hemolysis of red blood cells; and (5) eventual subsiding of the interstitial edema in surviving rats with a concomitant development of emphysema.
The dynamics of secondary vacuole movement is studied in living hair cells of Tradescantia virginiana. The pattern of movement of these vacuoles is found to be similar to that described by the author previously for organelles in cultured cells. Evidence is presented in support of the thesis that the occurrence and dynamics of secondary vacuoles is a common phenomenon for plant cells.
The anatomy of the intratemporal part of the vestibular nerve in man, and the possible age related degenerative changes in the nerve were studied. The form and structure of the vestibular ganglion was studied with the light microscope. A numerical analysis of the vestibular nerve, and caliber spectra of the myelinated fibers in the vestibular nerve branches were studied in individuals of varying ages. It was found that the peripheral endings of the vestibular nerve form a complicated pattern inside the vestibular sensory epithelia. A detailed description of the sensory cells and their surface organelles is included.
Plant and certain animal embryos appear to be responsive to the gravity vector during early stages of development. The convection of particle sedimentation as the basis for the sensing of gravity is investigated using the cells of wheat seedlings, amphibian embryos, and mammals. Exploration of the mammalian cell for sedimenting particles reveals that their existence is unlikely, especially in the presence of a network of microtubules and microfilaments considered to be responsible for intracellular organization. Destruction of these structures renders the cell susceptible to accelerations several times g. Large dense particles, such as chromosomes, nucleoli, and cytoplasmic organelles are acted upon by forces much larger than that due to gravity, and their positions in the cell appear to be insensitive to gravity.
It is argued that the transition to an oxidizing atmosphere preceded the origin of eukaryotic cells, which in turn must have preceded the origin of metazoa. Moreover, the number of methods by which organisms can protect themselves from harmful UV radiation is sufficiently large to suggest that solar UV, even when the atmosphere was anaerobic, was not such as to control the distribution and diversification of life. An alternative explanation for the late and sudden appearance of metazoa in lower Cambrian sediments is proposed, which is related to the mechanisms by which fully mature eukaryotic cells probably originated. There was probably a protracted evolution of modern genetic systems based on mitosis in cells which acquired organelles (e.g., plastids and mitochondria) by hereditary endosymbiosis. The origin of hard parts underlies the Cambrian explosion of metazoans.
Studies about the origin of life are reviewed. The nonrandom organization of organelles is discussed from a structural and functional point of view. After postulating that the origin of biomacromolecules was not a random event, the paper develops the hypothesis that polypeptides and polynucleotides were formed on an inorganic template. Only information-containing structures can pass natural selection and develop through evolution.
The possibility is put forth that the mitochondrion did not originate from an endosymbiosis 1-2 billion years ago involving an aerobic bacterium. Rather, it arose by endosymbiosis in a much earlier anaerobic period and was initially a photosynthetic organelle analogous to the modern chloroplast. This suggestion arises from a reconsideration of the nature of endosymbiosis. It explains the remarkable diversity in mitochondrial information storage and processing systems.
A computer branching model is used to analyze cellular evolution. Attention is given to certain key amino acids and nucleotide residues (ferredoxin, 5s ribosomal RNA, and c-type cytochromes) because of their commonality over a wide variety of cell types. Each amino acid or nucleotide residue is a sequence in an inherited biological trait; and the branching method is employed to align sequences so that changes reflect substitution of one residue for another. Based on the computer analysis, the symbiotic theory of cellular evolution is considered the most probable. This theory holds that organelles, e.g., mitochondria and chloroplasts invaded larger bodies, e.g., bacteria, and combined functions to form eucaryotic cells.
The past, present and expected future accomplishments and limitations of plant cell and tissue culture are reviewed. Consideration is given to the pioneering insights of Haberlandt in 1902, the development of culture techniques, and past work on cell division, cell and tissue growth and development, somatic embryogenesis, and metabolism and respiration. Current activity in culture media and technique development for plant regions, organs, tissues, cells, protoplasts, organelles and embryos, totipotency, somatic embryogenesis and clonal propagation under normal and space conditions, biochemical potentialities, and genetic engineering is surveyed. Prospects for the investigation of the induced control of somatic cell division, the division of isolated protoplasts, the improvement of haploid cell cultures, liquid cultures for somatic embryogenesis, and the genetic control of development are outlined.
An experiment to study the effect of weightlessness on lymphocyte proliferation to detect possible alteration of the cells responsible for the immune response during long-duration space flights is described. Human lymphocytes in culture medium will be delivered shortly before launch in an incubator which will be kept at 37C. Mitogen will be added to the culture. A control without mitogen will be run in parallel. After 70 hours of incubation, radioactive thymidine will be added. After two hours, cellular activity will be stopped by fixation and incubator power switched off. Later, the amount of incorporated thymidine will be determined and the cell morphology and the distribution of cell organelles will be investigated.
Aqueous phase-separated polymer solutions can be used as support media for the partition of biological macromolecules, organelles and cells. Cell separations using the technique have proven to be extremely sensitive to cell surface properties but application of the systems are limited to cells or aggregates which do not significantly while the phases are settling. Partition in zero g in principle removes this limitation but an external driving force must be applied to induce the phases to separate since their density difference disappears. We have recently shown that an applied electric field can supply the necessary driving force. We are proposing to utilize the NASA FES to study field-driven phase separation and cell partition on the ground and in zero g to help define the separation/partition process, with the ultimate goal being to develop partition as a zero g cell separation technique.
The anatomy and fine structure of roots from oat and mung bean seedlings grown under hypogravity conditions aboard NASA's Space Shuttle were examined and compared to those of roots from ground control plants grown under similar conditions. Oat roots from both sets of plants exhibited normal tissue organization and ultrastructural features, with the exception of cortex cell mitochondria, which characteristically showed a 'swollen' morphology. Flight-grown mung bean roots differed significantly from the controls in that root cap cells were somewhat disorganized and degraded in appearance, especially at the cap periphery. At the EM level, these cells exhibited a loss organelle integrity and a condensed cytoplasm. The potential significance of this finding for the putative gravity-sensing cap cells were noted.
Amyloplast DNA (apDNA), when subjected to digestion with restriction endonucleases, yields patterns nearly identical to that of DNA from mature pea chloroplasts (ctDNA). Southern transfers of apDNA and ctDNA, probed with the large subunit (LS) gene of ribulose-1,5-bisphosphate carboxylase (Rubisco), shows hybridization to the expected restriction fragments for both apDNA and ctDNA. However, Northern transfers of total RNA from chloroplasts and amyloplasts, probed again with the LS gene of Rubisco, shows that no detectable LS meggage is found in amyloplasts although LS expression in mature chloroplasts is high. Likewise, two dimensional polyacrylamide gel electrophoresis of etiolated gravisensitive pea tissue shows that both large and small subunits of Rubisco are conspicuously absent; however, in greening tissue these two constitute the major soluble proteins. These findings suggest that although the informational content of these two organelle types is equivalent, gene expression is quite different and is presumably under nuclear control.
Gravity affects biology at almost all levels above that of the cell organelle. Attention is presently given to progress made in the understanding of gravitational effects through studies employing centrifuges, clinostats, inverted preparations, linear devices, water immersion, free fall, and short- and long-term spaceflight. The cardiovascular changes which cause malaise and illness during the first few days of extended space missions are the direct result of fluid translocation from the lower extremities. Upon reentry, there is hypovolumnia and a cardiovascular deconditioning that can include tachycardia, changes in arterial blood pressure, narrow pulse pressure, and syncope. Attention is also given to NASA's gravitational physiology reseach program.
Prokaryotic and eukaryotic evolutionary trees are developed from protein and nucleic-acid sequences by the methods of numerical taxonomy. Trees are presented for bacterial ferredoxins, 5S ribosomal RNA, c-type cytochromes , cytochromes c2 and c', and 5.8S ribosomal RNA; the implications for early evolution are discussed; and a composite tree showing the branching of the anaerobes, aerobes, archaebacteria, and eukaryotes is shown. Single lines are found for all oxygen-evolving photosynthetic forms and for the salt-loving and high-temperature forms of archaebacteria. It is argued that the eukaryote mitochondria, chloroplasts, and cytoplasmic host material are descended from free-living prokaryotes that formed symbiotic associations, with more than one symbiotic event involved in the evolution of each organelle.
Antimonate staining procedures were used to detect calcium redistribution changes in corn roots. Results show that an asymmetric redistribution of Ca is induced by a gravitropic stimulus in roots as it is in shoots. Since this response occurs within 10 min, at least 5 min before any visible bending, it could play a role in the regulation of root gravitropism. Two different general approaches were used to localize calmodulin in plant tissue: radioimmunoassay of its content in tissue and in purified subcellular organelles, and immunocytochemical detection of it in roots and coleoptiles. Radioimmunoassay results indicate that calmodulin is present in large quantities in pllant cells and that it is specifically associated with mitochondria, etioplasts and nuclei. An assayed of an extract of soluble wall proteins revealed that over 1% of these proteins was calmodulin. Controls indicate that this calmodulin is not cytoplasmic in origin. A reaction product from anti-calmodulin was found mainly in the root cap cells, moderately in metazylem elements, in some cells in the stele surrounding metaxylem elements and in cortical cells.
The overall process of gravity sensing and response processes in plants may be divided conveniently into at least four components or stages: Stimulus susception (a physical event, characteristically the input to the G receptor system of environmental information about the G force magnitude, its vector direction, or both); information perception (an influence of susception on some biological structure or process that can be described as the transformation of environmental information into a biologicallly meaningful change); information transport (the export, if required, of an influence (often chemical) to cells and organs other than those at the sensor location); and biological response (almost always (in plants) a growth change of some kind). Some analysts of the process identify, between information perception and information transport, an additional stage, transduction, which would emphasize the importance of a transformation from one form of information to another, for example from mechanical statolith displacement to an electric, chemical, or other alteration that was its indirect result. These four (or five) stages are temporally sequential. Even if all that occurs at each stage can not be confidently identified, it seems evident that during transduction and transport, matters dealt with are found relatively late in the information flow rather than at the perception stage. As more and more is learned about the roles played by plant hormones which condition the G responses, the mechanism(s) of perception which should be are not necessarily better understood. However, if by asking the right questions and being lucky with experiments perhaps the discovery of how some process (such as sedimentation of protoplasmic organelles) dictates what happens down stream in the information flow sequence may be made.