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At least 109 records · Page 6

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NASA NTRS1972

Prebiotic phosphorylation of thymidine at 65 C in simulated desert conditions.

The phosphorylation of thymidine is described for a variety of conditions at 65 C to demonstrate that the reaction could readily take place in deserts at the present time. This might be used as an indication that urea-phosphate mixtures could have been important as phosphorylating agents on the primitive earth. Reaction products were identified by comparing their chromatographic and electrophoretic mobilities with those of authentic materials and by enzymatic degradation. The results show that good yields of nucleotides are obtained when nucleosides are heated with urea-phospate mixtures at 65 C. Reactions proceed more rapidly at moderate humidities than in a stream of dry nitrogen. Occasional wetting results in even faster and more extensive reactions. There was no reaction for a mixture of urea and trimetaphosphate.

Bishop, M. J.↗

NASA NTRS1973

Method of detecting and counting bacteria in body fluids

A novel method is reported for determining bacterial levels in urine samples, which method depends on the quantitative determination of bacterial adenosine triphosphate (ATP) in the presence of non-bacterial ATP. After the removal of non-bacterial ATP, the bacterial ATP is released by cell rupture and is measured by an enzymatic bioluminescent assay using an enzyme obtained from the firefly.

Chappelle, E. W.↗

NASA NTRS1973

Glycolytic intermediates and adenosine phosphates in rat liver at high altitude /3,800 m/.

Liver tissue obtained from adult rats exposed to 3800 m altitude for intervals ranging from 1.5 hr to 63 days was examined by enzymatic analysis. During the first 3 hr of exposure, an immediate decrease in rephosphorylation of high-energy phosphates led to reduced glycogenesis and eventual pileup of AMP, pyruvate, fructose 1,6-diphosphate, glucose 6-phosphate, and glucose. This was accompanied by a reduction of pentose phosphate pathway activity. After 3 to 6 hr, a secondary adjustment of substrate concentrations occurred along with the apparent facilitation of phosphofructokinase. This secondary adjustment appears to increase anaerobic production of ATP and represents a significant intracellular contribution to the acclimatization process at high altitude.

Cipriano, L. F.↗

NASA NTRS1974

The prebiotic synthesis of deoxythymidine oligonucleotides

Deoxythymidine 5 prime-triphosphate in the presence of deoxythymidine 5 prime-phosphate, cyanamide and 4-amino-5-imidazole carboxamide polymerizes under drying conditions at moderate temperatures (60 to 90 C) to yield oligonucleotides of up to four units in length. Enzymatic analysis indicated that the majority of these oligomers contained natural 3 prime-5 prime phosphodiester bonds. This reaction offers a possible method for the formation of deoxyoligonucleotides under primitive earth conditions.

Stephen-Sherwood, E.↗

NASA NTRS1976

Method of detecting and counting bacteria

An improved method is provided for determining bacterial levels, especially in samples of aqueous physiological fluids. The method depends on the quantitative determination of bacterial adenosine triphosphate (ATP) in the presence of nonbacterial ATP. The bacterial ATP is released by cell rupture and is measured by an enzymatic bioluminescent assay. A concentration technique is included to make the method more sensitive. It is particularly useful where the fluid to be measured contains an unknown or low bacteria count.

Picciolo, G. L.↗

NASA NTRS1976

Streaming potential measurements of biosurfaces

A technique based on the measurement of streaming potentials has been developed to evaluate the electrokinetic region of the cell periphery. This approach is feasible for cell lines propagated in in-vitro cell cultures in monolayer form. The advantage of this system is that cells may be evaluated in the living state atttached to a substrate; it is not necessary to subject the cells to enzymatic, chemical, or mechanical trauma required to obtain monodisperse suspensions which are then normally evaluated by microelectrophoresis. In this manner, it should be possible to study the influence of substrate and environmental factors on the charge density and potential at the cell periphery. The apparatus and procedure are described as well as some results concerning the electrokinetic potential of borosilicate capillaries as a function of ionic strength, pH, and temperature. The effect that turbulence and entrance flow conditions have on accurate streaming-potential measurements is discussed. The electrokinetic potential of BALB/c 3T12 fibroblasts has been quantified as a function of pH, ionic strength, glutaraldehyde fixation, and Giemsa staining.

Van Wagenen, R. A.↗

NASA NTRS1976

Space biosynthesis systems

Model reactions based on chemical, enzymatic or cellular conversion of D glucose into d gluconic acid are designed to unequivocally define the advantages of microgravity on reaction mechanisms, mass-transfers and separation of organic chemicals and to serve as procedures to test the performance characteristics of space bioprocessing equipment.

Nyiri, L. K.↗

NASA NTRS1980

The effects of adrenalectomy and corticsteroid injection on the fibrinolytic activity of complex heparin compounds in the blood during immobilization

Total non-enzymatic fibrinolytic activity in the blood of rats increased three times in response to stress caused by 30 minute immobilization, and the activity of epinephrine-heparin complex increased nine times. In adrenalectomized animals, which showed a weak response to the same stress, intraperitoneal injection of hydrocortisone 30 minutes prior to immobilization normalized the response. Obtained results indicate that adrenalectomy leads to sharp reduction of heparin complexing with thromogenic proteins and epinephrine, while substitution therapy with hydrocortisone restores anticoagulation system function.

Kudryashov, B. A.↗

NASA NTRS1980

Energy status and oxidation reduction status in rat liver at high altitude /3.8 km/

Adult male rats were exposed to 3.8-km altitude for intervals ranging from 1 h-60 d. Liver samples were taken under light ether anesthesia and were examined by enzymatic analyses. Within 1-6 h of hypoxic exposure, ATP levels decreased while ADP and AMP levels increased, producing a fall in calculated ATP/ADP and adenylate charge ratios. Concurrently, lactate/pyruvate and alpha-glycerophosphate/dihydroxyacetone phosphate ratios increased markedly. Direct measurements of cellular pyridine nucleotides indicated increased NADH/NAD and NADPH/NADP ratios. Levels of total adenosine phosphates and pyridine nucleotides decreased in a significant accompanying response. Many metabolite levels and calculated ratios returned to near-normal values within 1 week of exposure, indicating secondary intracellular adjustments to hypoxic stress; however, persistence of that stress is reflected in lactate concentrations and both substrate redox ratios. Results support and explore concepts that increased oxidation-reduction status and decreased energy status are primary events during hypoxia.

Reed, R. D.↗

NASA NTRS1980

Unification of some biochemical methods of research in the pre- and post-flight periods

The biochemical methods for determination of various parameters and factors during pre- and post-flight periods, as used by American and Soviet teams dealing with space flight medicine are compared. The emphasis is on the exchange of information on the study of the blood and urine content of space travelers before and after space flight. A series of electrolytic, enzymatic, and hormonal factors is discussed.

Tigranyan, R. A.↗

NASA NTRS1982

Analysis of volatile metabolites in biological fluids as indicators of prodromal disease condition

The volatile profile cannot be defined as a single class of substances, rather it is a broad spectrum of materials of different polarities characterized by having a boiling-point in the low to medium range (up to approximately 300 C) and the fact that the compounds are suitable for gas chromatography without derivatization. The organic volatile profiles are very complex mixtures of metabolic byproducts, intermediates, and terminal products of enzymatic degradations composed mainly of alcohols, ketones, aldehydes, pyrazines, sulfides, isothiocyanates, pyrroles, and furans. The concentration of organic volatiles in biological fluids covers a wide range with many important components present at trace levels. The complexity of the organic volatile fraction requires the use of capillary columns for their separation.

Zlatkis, A.↗

NASA NTRS1983

Effect of denervation and reinnervation on oxidation of 6-(C-14) glucose by rat skeletal muscle homogenates

The effects of denervation and reinnervation of the rat extensor digitorum longus muscle on the oxidation of 6-(C-14) glucose to (C-14)O2 is investigated. Results show that the rate of (C-14)O2 production decreased dramatically following denervation and the decrease became significant 20 days after nerve section. The changes which occurred prior to day 20 apparently reflected the decline of muscle mass. The decreased (C-14)O2 production was found to be due to reduced capacity of the enzymatic system, while there was no change in the apparent affinity for glucose. Results of mixing experiments showed that the loss of oxidative capacity following denervation is not caused by the production of soluble inhibitors by degenerating muscle. Measurements of the (C-14)O2 revealed that oxidative metabolism recovered during reinnervation. The specific activity in reinnervated muscles displayed an 'overshoot' of approximately 50 percent, which returned to control levels by day 60. The time-course of the denervation-mediated change indicates that altered oxidative capacity is secondary to events that initiate dennervation changes in muscle, although diminished oxidative capacity may be of considerable metabolic significance in denervated muscle.

Dubois, D. C.↗

NASA NTRS1983

Chemical evolution and the origin of life

A review is presented of recent advances made in the understanding of the formation of carbon compounds in the universe and the occurrence of processes of chemical evolution. Topics discussed include the principle of evolutionary continuity, evolution as a fundamental principle of the physical universe, the nuclear synthesis of biogenic elements, organic cosmochemistry and interstellar molecules, the solar nebula and the solar system in chemical evolution, the giant planets and Titan in chemical evolution, and comets and their interaction with the earth. Also examined are carbonaceous chondrites, environment of the primitive earth, energy sources available on the primitive earth, the synthesis of biochemical monomers and oligomers, the abiotic transcription of nucleotides, unified prebiotic and enzymatic mechanisms, phospholipids and membranes, and protobiological evolution.

Oro, J.↗

NASA NTRS1984

Cellular localization of Na(+), K(+)-ATPase in the mammalian vestibular system

Two different, but complementary, procedures for cellular localization of Na+, K+-ATPase in the guinea pig vestibular system were employed. One of these techniques, devised by Stirling, depends upon the well documented ability of the specific inhibitor ouabain to bind selectively to Na+,K+-ATPase, blocking catalytic activity. Microdisected vestibular tissues are incubated with tritium-labelled (3H-) ouabain, and regions with a high concentration of Na+,K+-ATPase are subsequently identified by light microscope autoradiography. A second method, originated by Ernst, detects inorganic phosphate released from an artificial substrate (nitrophenyl phosphate) by catalytic activity of the enzyme. In the presence of strontium ion, phosphate is precipitated near regions of high activity, then converted to a product which may finally be visualized in the electron microscope. This cytochemical enzymatic reaction is inhibited by ouabain.

Kerr, T. P.↗

NASA NTRS1986

Oxygen and the evolution of metabolic pathways

While a considerable amount of evidence has been accumulated about the history of oxygen on this planet, little is known about the relative amounts to which primitive cells might have been exposed. One clue may be found in the metabolic pathways of extant microorganisms. While eucaryotes are principally aerobic organisms, a number are capable of anaerobic growth by fermentation. One such eucaryotic microorganism, Saccharomyces cerevisiae, will grow in the complete absence of oxygen when supplemented with unsaturated fatty acid and sterol. Oxygen-requiring enzymes are involved in the synthesis of both of these compounds. Studies have demonstrated that the oxidative desaturation of palmitic acid and the conversion of squalene to sterols occur in the range of 10-(3) to 10(-2) PAL. Thus, if the oxygen requirements of these enzymatic processes are an indication, eucaryotes might be more primitive than anticipated from the microfossil record. Results of studies on the oxygen requirements for sterol and unsaturated fatty acid synthesis in a more primitive procaryotic system are also discussed.

Jahnke, L. L.↗

NASA NTRS1985

Potential for utilization of algal biomass for components of the diet in CELSS

Techniques which eliminate or reduce the undesirable cell components of algae and enhance the potential nutritional and organoleptic acceptability of algae products are studied. The cell walls, nucleic acids, and pigments and lipids of the green algae Scenedesmus obiliquus need to be removed. The procedures for determining the composition of proteins, pigments and lipids, and moisture and ash are described. Chemical, enzymatic, and physical methods of removing the cell wall to make the algae digestable are analyzed; a homogenization technique is utilized. The problems encountered if algae nucleic acids are ingested directly are discussed; the reduction of DNA and RNA by applying extracellular DNase and RNase to the nucleic acids is examined. The color and flavor of the algae are enhanced with the extraction of pigments and lipids from the algae protein concentration.

Kamarei, A. R.↗

NASA NTRS1987

Electrophoretic enzyme analysis of North American and eastern Asian populations of Agastache sect. Agastache (Labiatae)

Genetic relationships among the seven species of Agastache sect. Agastache common in North America and the one found in eastern Asia were assessed using starch-gel electrophoresis of twelve enzymatic proteins. Nei's (1976) genetic distance and identity values, calculated among the 32 populations used in this study, partitioned the Agastache section into four discrete groups: (1) A. nepetoides (eastern North America), (2) A. scrophulariifolia and A. foeniculum (eastern and central North America), (3) the four species of the western U.S. (A. urticifolia, A. occidentalis, A. parvifolia, and A. cusickii), and (4) A. rugosa (eastern Asia). The Asian Agastache, separated from its American congeners for over 12 million years, differed from American populations at only two (the IDH-1 and LAP-1 alleles) of the fifteen loci surveyed; these alleles were not found in any of the North American plants. Nei's genetic distances between the Asian and North American populations ranged from 0.2877 to 0.6734.

Vogelmann, James E.↗

NASA NTRS1987

Hydroxide decomposition of dimethylsulfoniopropionate to form dimethylsulfide

The kinetics of DMS production resulting from reaction of OH(-) with DMSP were investigated as a function of hydroxide concentration and temperature. The reaction was first-order with respect to DMSP and OH(-). The second order rate constant at 20+/-1 C is 0.0044/M/sec. The activation energy for this reaction is 14.4 kcal/mode. The investigation indicates that the rate of reaction of DMSP with OH(-) is very slow at the pH of seawater, suggesting that DMSP, which may be a major precursor of DMS in seawater, decomposes in the ocean by other mechanisms. A bacterium which produces DMS from DMSP quantitatively at rates many orders of magnitude higher than indicated by OH(-1) decomposition has been cultured, suggesting that enzymatic processes accelerate the production of DMS from DMSP in seawater.

Dacey, John W. H.↗