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At least 73 records · Page 4

Experiment K304: Studies of specific hepatic enzymes and liver constituents involved in the conversion of carbohydrates to lipids in rats exposed to prolonged space flight

The effects of space flight on the activities of 26 enzymes concerned with carbohydrate and lipid metabolism in hepatic tissue taken from male Wistar rats are investigated. These activities were measured in the various hepatic cell compartments, i.e., cytosol, mitochondria and microsomes. In addition, the levels of glycogen, total lipids, phospholipids, triglycerides, cholesterol, cholesterol esters, and the fatty acid composition of the rat livers were also examined and quantified. A similar group of ground-based rats treated in an identical manner served as controls. Both flight and synchronous control rats were sacrificed at three time intervals: R+0, 7-11 hours after recovery; R+6, after 6 days; R+6(S), after 6 days (having undergone 2-5 hour periods of fixed stress in a "backupward" position on days 0, 3, 4, 5 and 6) and R+29, after 29 days post-flight. Although most of the enzyme activities and the amounts of liver constituents studied were unaffected by the period of weightlessness, some significant differences were observed.

Abraham, S.↗

Symbiosis in cell evolution: Life and its environment on the early earth

The book treats cell evolution from the viewpoint of the serial endosymbiosis theory of the origin of organelles. Following a brief outline of the symbiotic theory, which holds that eukaryotes evolved by the association of free-living bacteria with a host prokaryote, the diversity of life is considered, and five kingdoms of organisms are distinguished: the prokaryotic Monera and the eukaryotic Protoctista, Animalia, Fungi and Plantae. Symbiotic and traditional direct filiation theories of cell evolution are compared. Recent observations of cell structure and biochemistry are reviewed in relation to early cell evolution, with attention given to the geological context for the origin of eukaryotic cells, the origin of major bacterial anaerobic pathways, the relationship between aerobic metabolism and atmospheric oxygen, criteria for distinguishing symbiotic organelles from those that originated by differentiation, and the major classes of eukaryotic organelles: mitochondria, cilia, microtubules, the mitotic and meiotic apparatuses, and pastids. Cell evolution during the Phanerozoic is also discussed with emphasis on the effects of life on the biosphere

Margulis, L.↗

Ultrastructure of meristem and root cap of pea seedlings under spaceflight conditions

Data of electron microscopic analysis of meristem and root cap of pea seedlings grown aboard the Salyut-6 orbital research station in the Oazis apparatus and in the laboratory are presented. The main morphological and anatomical characteristics of the test and control plants are shown to be similar. At the same time, some differences are found in the structural and functional organization of the experimental cells as compared to the controls. They concern first of all the plastic apparatus, mitochondria and Golgi apparatus. It is assumed that cell function for certain periods of weightlessness on the whole ensures execution of the cytodifferentiation programs genetically determined on the Earth. Biochemical and physiological processes vary rather markedly due to lack of initially rigorous determination.

Sytnyk, K. M.↗

A comparative study of monocot and dicot root development in normal /earth/ and hypogravity /space/ environments

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.

Slocum, R. D.↗

The Plasma Membrane Calcium Pump

Three aspect of cellular calcium metabolism in animal cells was discussed including the importance of the plasma membrane in calcium homeostasis, experiments dealing with the actual mechanism of the calcium pump, and the function of the pump in relationship to the mitochondria and to the function of calmodulin in the intact cell.

Rasmussen, H.↗

Evolutionary connections of biological kingdoms based on protein and nucleic acid sequence evidence

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.

Dayhoff, M. O.↗

Calcium and Calmodulin Localization in Gravitropically-responding Plant Organs

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.

Roux, S. J.↗

Distribution and abundance of organic thiols

The role of glutathione (GSH) in protecting against the toxicity of oxygen and oxygen by products is well established for all eukaryotes studied except Entamoeba histolytica which lacks mitochrondria, chloroplasts, and microtubules. The GSH is not universal among prokaryotes. Entamoeba histolytica does not produce GSH or key enzymes of GSH metabolism. A general method of thiol analysis based upon fluorescent labeling with monobromobimane and HPLC separation of the resulting thiol derivatives was developed to determine the occurrence of GSH and other low molecular weight thiols in bacteria. Glutathione is the major thiol in cyanobacteria and in most bacteria closely related to the purple photosynthetic bacteria, but GSH was not found in archaebacteria, green bacteria, or GRAM positive bacteria. It suggested that glutathione metabolism was incorporated into eukaryotes at the time that mitochondria and chloroplasts were acquired by endosymbiosis. In Gram positive aerobes, coenzyme A occurs at millimolar levels and CoA disulfide reductases are identified. The CoA, rather than glutathione, may function in the oxygen detoxification processes of these organisms.

Fahey, R.↗

Higher-order structure of rRNA

A comparative search for phylogenetically covarying basepair replacements within potential helices has been the only reliable method to determine the correct secondary structure of the 3 rRNAs, 5S, 16S, and 23S. The analysis of 16S from a wide phylogenetic spectrum, that includes various branches of the eubacteria, archaebacteria, eucaryotes, in addition to the mitochondria and chloroplast, is beginning to reveal the constraints on the secondary structures of these rRNAs. Based on the success of this analysis, and the assumption that higher order structure will also be phylogenetically conserved, a comparative search was initiated for positions that show co-variation not involved in secondary structure helices. From a list of potential higher order interactions within 16S rRNA, two higher-order interactions are presented. The first of these interactions involves positions 570 and 866. Based on the extent of phylogenetic covariation between these positions while maintaining Watson-Crick pairing, this higher-order interaction is considered proven. The other interaction involves a minimum of six positions between the 1400 and 1500 regions of the 16S rRNA. Although these patterns of covariation are not as striking as the 570/866 interaction, the fact that they all exist in an anti-parallel fashion and that experimental methods previously implicated these two regions of the molecule in tRNA function suggests that these interactions be given serious consideration.

Gutell, R. R.↗

Is the ATPase from halobacterium saccharovorum an evolutionary relic?

The ATP Synthase Complex present in the membranes of mitochondria, chloroplasts or bacteria is composed of 2 sectors: FO, an integral membrane protein consisting of 3 subunits mediating proton translocation across the membrane and F1, the catalytic component composed of 5 non-identical subunits. The apparent early origin of the ATP Synthase Complex, as implied by its ubiquitous distribution, seems inconsistent with its structural and functional complexity and raises the question if simpler versions of the ATP Synthase exist. Such an ATP Synthase has been searched for in various Archaebacteria. A purified halobacterial ATPase activity which possesses certain properties consistent with those of an ATP Synthase but which has a different subunit structure is described.

Hochstein, L. I.↗

Phylogenetic origins of the plant mitochondrion based on a comparative analysis of 5S ribosomal RNA sequences

The complete nucleotide sequences of 5S ribosomal RNAs from Rhodocyclus gelatinosa, Rhodobacter sphaeroides, and Pseudomonas cepacia were determined. Comparisons of these 5S RNA sequences show that rather than being phylogenetically related to one another, the two photosynthetic bacterial 5S RNAs share more sequence and signature homology with the RNAs of two nonphotosynthetic strains. Rhodobacter sphaeroides is specifically related to Paracoccus denitrificans and Rc. gelatinosa is related to Ps. cepacia. These results support earlier 16S ribosomal RNA studies and add two important groups to the 5S RNA data base. Unique 5S RNA structural features previously found in P. denitrificans are present also in the 5S RNA of Rb. sphaeroides; these provide the basis for subdivisional signatures. The immediate consequence of obtaining these new sequences is that it is possible to clarify the phylogenetic origins of the plant mitochondrion. In particular, a close phylogenetic relationship is found between the plant mitochondria and members of the alpha subdivision of the purple photosynthetic bacteria, namely, Rb. sphaeroides, P. denitrificans, and Rhodospirillum rubrum.

Villanueva, E.↗

Axoglial contacts in the area postrema of the cat - An ultrastructural study

Axoglial contacts were observed in an ultrastructural study of the area postrema of the cat. According to the disposition of the electron-dense projections attached to the adjoining membranes these contacts were classified as symmetrical or asymmetrical. The axon profiles contained aggregations of clear vesicles randomly distributed or grouped in clusters adjacent to the electron-dense projections. Dense core vesicles were occasionally seen. The neuroglial profiles were either astrocytic or ependymoglial in nature. The astrocytes showed a clear cytoplasm, polymorphous vesicles, mitochondria, glycogen granules, and bundles of filaments. The ependymal cells, in contrast, had a more electron-dense and granular appearance, tubular structures, irregular vesicular formations, profiles of smooth reticuloendoplasm, and filaments grouped in bundles or isolated in the cytoplasm. The possibility that these contacts might play a role in the chemical transfer from neurons to glial cells is discussed on the basis of existing biochemical data.

Damelio, Fernando E.↗

Experiment K-6-09. Morphological and biochemical investigation of microgravity-induced nerve and muscle breakdown. Part 1: Investigation of nerve and muscle breakdown during spaceflight; Part 2: Biochemical analysis of EDL and PLT muscles

The present findings on rat hindlimb muscles suggest that skeletal muscle weakness induced by prolonged spaceflight can result from a combination of muscle fiber atrophy, muscle fiber segmental necrosis, degeneration of motor nerve terminals and destruction of microcirculatory vessels. Damage was confined to the red adductor longus (AL) and soleus muscles. The midbelly region of the AL muscle had more segmental necrosis and edema than the ends. Macrophages and neutrophils were the major mononucleated cells infiltrating and phagocytosing the cellular debris. Toluidine blue-positive mast cells were significantly decreased in Flight AL muscles compared to controls; this indicated that degranulation of mast cells contributed to tissue edema. Increased ubiquitination of disrupted myofibrils may have promoted myofilament degradation. Overall, mitochondria content and SDH activity were normal, except for a decrease in the subsarcolemmal region. The myofibrillar ATPase activity shifted toward the fast type in the Flight AL muscles. Some of the pathological changes may have occurred or been exacerbated during the 2 day postflight period of readaptation to terrestrial gravity. While simple atrophy should be reversible by exercise, restoration of pathological changes depends upon complex processes of regeneration by stem cells. Initial signs of muscle and nerve fiber regeneration were detected. Even though regeneration proceeds on Earth, the space environment may inhibit repair and cause progressive irreversible deterioration during long term missions. Muscles obtained from Flight rats sacrificed immediately (within a few hours) after landing are needed to distinguish inflight changes from postflight readaptation.

Riley, D. A.↗

Experiment K-6-13. Morphological and biochemical examination of heart tissue. Part 1: Effects of microgravity on the myocardial fine structure of rats flown on Cosmos 1887. Ultrastructure studies. Part 2: Cellular distribution of cyclic ampdependent protein kinase regulatory subunits in heart muscle of rats flown on Cosmos 1887

The left ventricle of hearts from rats flown on the Cosmos 1887 biosatellite for 12.5 days was compared to the same tissue of synchronous and vivarium control animals maintained in a ground based laboratory. The volume density of the mitochondria in the myocardium of the space-flown animals was statistically less (p equal less than 0.01) than that of the synchronous or vivarium control rats. Exposure to microgravity resulted in a certain degree of myocardial degeneration manifested in mitochondrial changes and accumulation of myeloid bodies. Generalized myofibrillar edema was also observed.

Philpott, D. E.↗

Rat limb unloading - Soleus histochemistry, ultrastructure, and electromyography

The effects of hindlimb unloading on rat-soleus histochemisty, ultrastructure, and electromyogram (EMG) activity were investigated. It was found that, after 14 days of tail suspension, the area of type I and type IIa muscle fibers decreased by 63 and 47 percent, respectively, mainly due to the degradation of subsarcolemmal mitochondria and myofibrils. After 10 days, 3 percent of type IIa fibers exhibited segmental necrosis. After four days, video monitoring revealed abnormal plantar flexion of the hindfeet, which shortened the soleus working range. The EMG activity shifted from tonic to phasic, and aggregate activity decreased drastically after only seven days. The results indictate that the pathological changes in the soleus resulted from unloaded contractions, reduced use, compromised blood flow, and shortened working length.

Riley, D. A.↗

Symbiosis and the origin of eukaryotic motility

Ongoing work to test the hypothesis of the origin of eukaryotic cell organelles by microbial symbioses is discussed. Because of the widespread acceptance of the serial endosymbiotic theory (SET) of the origin of plastids and mitochondria, the idea of the symbiotic origin of the centrioles and axonemes for spirochete bacteria motility symbiosis was tested. Intracellular microtubular systems are purported to derive from symbiotic associations between ancestral eukaryotic cells and motile bacteria. Four lines of approach to this problem are being pursued: (1) cloning the gene of a tubulin-like protein discovered in Spirocheata bajacaliforniesis; (2) seeking axoneme proteins in spirochets by antibody cross-reaction; (3) attempting to cultivate larger, free-living spirochetes; and (4) studying in detail spirochetes (e.g., Cristispira) symbiotic with marine animals. Other aspects of the investigation are presented.

Margulis, L.↗

Cardiac morphology after conditions of microgravity during Cosmos 2044

Light- and electron-microscopic studies were performed on cardiac muscle from rats flown on Cosmos 2044 and from four control groups. Average cross-sectional area of myofibers was measured by video analysis of the light-microscopic images of papillary and ventricular muscle samples from all animals. This cross-sectional area was significantly decreased in flight rats (P = 0.03) compared with synchronous controls. Additional findings at the electron microscopic level consistent with this atrophy were obtained by stereological analysis and optical diffraction analysis of papillary muscle samples. Slightly higher mitochondrial volume density values and mitochondria-to-myofibril ratios as well as normal A-band spacings (d1,0) and Z-band spacings of myofibrils were observed in the tail-suspension and flight groups. General morphological features similar to those in ventricular samples from the previous Cosmos 1887 flight were observed.

Goldstein, Margaret A.↗

Muscle mitochondrial density after exhaustive exercise in dogs - Prolonged restricted activity and retraining

The effect of exhaustive treadmill exercise on mitochondrial density (MD) and ultrastructural changes in quadriceps femoris muscle was studied in 7 normal, healthy, male mongrel dogs before and after restricted activity (RA) and following a subsequent 2-month exercise retraining period. Mean time to exhaustion in the 2-month group decreased from 177 +/- 11 min before to 90 +/- 16 min after RA; retraining increased tolerance to 219 +/- 36 min above the pre-RA and 143 percent above the post-RA time. Post-RA exhaustion time in the 5-months group was 25 and 45 min. Muscle samples taken after RA showed abnormalities indicative of degeneration, which were reversed by retraining. Resting MD decreased from a control level of 27.8 percent to 14.7 percent and 16.3 percent, and was restored to 27.1 percent after retraining. Exhaustive exercise caused an increase in MD under control conditions and after RA, but not following retraining. Disruption of mitochondria after exercise was evident after 5-month confinement. Factors causing mitochondrial changes and eventually their disruption during exercise after restricted activity are not related as much to the state of fatigue as to the pre-exercise quality of the muscle modified by disease or training.

Nazar, K.↗