Catabolism of Tritiated Thymidine by Aquatic Microbial Communities and Incorporation of Tritium into RNA and Protein

The incorporation of tritiated thymidine by five microbial ecosystems and the distribution of tritium into DNA, RNA, and protein were determined. All microbial assemblages tested exhibited significant labeling of RNA and protein (i.e., nonspecific labeling), as determined by differential acid-base hydrolysis. Nonspecific labeling was greatest in sediment samples, for which ≥95% of the tritium was recovered with the RNA and protein fractions. The percentage of tritium recovered in the DNA fraction ranged from 15 to 38% of the total labeled macromolecules recovered. Nonspecific labeling was independent of both incubation time and thymidine concentration over very wide ranges. Four different RNA hydrolysis reagents (KOH, NaOH, piperidine, and enzymes) solubilized tritium from cold trichloroacetic acid precipitates. High-pressure liquid chromatography separation of piperidine hydrolysates followed by measurement of isolated monophosphates confirmed the labeling of RNA and indicated that tritium was recovered primarily in CMP and AMP residues. We also evaluated the specificity of [2-³H]adenine incorporation into adenylate residues in both RNA and DNA in parallel with the [³H]thymidine experiments and compared the degree of nonspecific labeling by [³H]adenine with that derived from [³H]thymidine. Rapid catabolism of tritiated thymidine was evaluated by determining the disappearance of tritiated thymidine from the incubation medium and the appearance of degradation products by high-pressure liquid chromatography separation of the cell-free medium. Degradation product formation, including that of both volatile and nonvolatile compounds, was much greater than the rate of incorporation of tritium into stable macromolecules. The standard degradation pathway for thymidine coupled with utilization of Krebs cycle intermediates for the biosynthesis of amino acids, purines, and pyrimidines readily accounts for the observed nonspecific labeling in environmental samples.     

A note on the structure of tail hairs from a pygmy hippopotamus (Choeropsis liberiensis)

The pygmy hippopotamus (Choeropsis liberiensis), like the Nile hippopotamus (Hippopotamus amphibius), defecates by backing into vertical objects while making a series of rapid, propellerlike tail movements that spread a mixture of urine and feces in a wide swath. Split hairs from the distal ventral surface of the pygmy hippopotamus tail were studied with the scanning electron microscope to determine whether the splitting was a normal character of the hair or was due to damage. The results suggest that splitting is a normal feature of the hair that may facilitate the dispersal of urine and feces.     

Plasminogen activator: The major secreted neutral protease of cultured skeletal muscle cells

Clonal mouse skeletal muscle cells which differentiate in culture and from synpases with neuronal cells were found to secrete high levels of protease activity as measured with an ¹²⁵I-fibrin assay. The secreted proteolytic activity was more than 90% dependent upon the presence of plasminogen in the medium, and had a pH optimum at 7 to 8. This activity was not inhibited by n-ethylmaleimide, pepstatin, EDTA, or EGTA. At millimolar concentrations, greater than 90% inhibition was obtained with either soybean typsin inhibitor, epsilon aminocaproic acid, Trasylol, or leupeptin. Almost complete inhibition occured with 1 mM diisopropylfluorophosphate suggesting the presence of a serine residue at the catalytic site. In contrast to the high levels of secreted activity, a lower steady-state level of cell-associated protease activity was detected in cell lysates. The high level of plasminogen activator secreted into the medium of cultured muscle cells suggests a role for such extracellular protease activity in myogenesis during development and remodeling following muscle injury. Such information may be useful in understanding the initial degeneration of neuromusclar contacts in experimental and pathologic denervation.     

Carbon Balance Studies of Glucose Metabolism in Rat Cerebral Cortical Synaptosomes

Synaptosomes were isolated from rat cerebral cortex and incubated with [U-14C]-, [1-14C]- or [6-14C]glucose. Glucose utilization and the metabolic partitioning of glucose carbon in products were determined by isotopic methods. From the data obtained a carbon balance was constructed, showing lactate to be the main product of glucose metabolism, followed by CO2, amino acids and pyruvate. Measuring the release of 14CO2 from glucose labelled in three different positions allowed the construction of a flow diagram of glucose carbon atoms in synaptosomes, which provides information about the contribution of the various pathways of glucose metabolism. Some 2% of glucose utilized was calculated to be degraded via the pentose phosphate pathway. Addition of chlorpromazine, imipramine or haloperidol at concentrations of 10(-5) M reduced glucose utilisation by 30% without changing the distribution pattern of radioactivity in the various products.     

Der informationsgehalt des intrazellularwassers und die informations-speicherung in der embryo- und Phylogenese

The nucleo-free erythrocyte is presented as the simplest differentiated cell whose energy-exchange has the single purpose of preserving the cell structure; this structure is based on the regular quasi-crystalline state of intracellular water, which is expressed by the negative entropy(-S _W )and the temperature T _W which is higher than the temperature of the extracellular water T. The information content of intracellular water, J _W is proportional to the temperature difference: J _W T=T _W -T. The regular state of intracellular water is maintained by the basal metabolism that takes place in the membrane of the differentiated cell. The energy exchange, i.e., the absorption of free energy and the liberation of an equivalent amount of heat, occurs in the form of work cycles of the enzym-water-complexes in the cell membrane. The differentiated cell of the multicellular animal organism is the result of embryogenetic processes accompanied by heat-liberation. The specific heat-liberation, i.e., the heat produced by a single cell, begins with a quasi-zero value of the fertilized egg cell and grows with acceleration to a maximum value at the end of embryogenesis. This process of acceleration of heat-liberation is caused by the entrance of the water from the outer medium into the embryonic cell; the water undergoes the phasechange fluid crystal with heat-liberation. The intracellular water within the embryonic cell becomes structurated; this is also accompanied by growing heatliberation. The thermodynamic characteristic of embryo genetic development is expressed by the principle of the maximum of velocity of entropy production of the cell at the end state of the process of differentiation. This principle applied to phylogenetics, leads to the formulation of the principle of accumulation of biological information: J(t). In the course of evolution living systems are able not only to store information of past generations, but also to create information: J(t)e^Intl where t is the time of phylogenesis.