Glucose-stimulated phosphorylation of yeast isocitrate lyase in vivo

Incorporation of 32P into Saccharomyces cerevisiae isocitrate lyase was observed after addition of glucose to a culture incubated with [32P]orthophosphoric acid. A band of 32P-labelled protein was coincident with the enzyme band when immunoprecipitates were subjected to SDS-PAGE and autoradiography. No label was found in the band corresponding to the isocitrate lyase when immunoprecipitation was done with a control pre-immune serum or in the presence of excess pure unlabelled enzyme. The incorporation of phosphate was associated with a decrease in enzyme activity. Phosphorylated isocitrate lyase was not proteolytically degraded when cells were cultured in mineral medium. The loss of protein antigenicity only took place when the yeast was grown in a complex medium containing glucose.     

Calmodulin Acts as an intermediary for the effects of calcium on gap junctions from crayfish lateral axons

Summary Lateral axons from the abdominal nerve cord of cray-fish were internally perfused with the calcium receptor calmodulin (CaM) in solutions with low (pCa>7.0) or high (pCa 5.5) calcium concentrations and studied electrophysiologically and morphologically. Results from these experiments show that when the internal solution contains calcium-activated calmodulin (Ca²⁺-CaM) the junctional resistance between the axons increases from control values of about 60 to 500–600 k in 60 min. In contrast, axons perfused with calmodulin in low calcium solutions maintain their junctional resistance at control levels during the 60-min perfusion. Similar results are obtained when only one or both coupled axons are perfused.The morphological study shows that in the perfused axons the axoplasmic organelles are replaced or grossly perturbed by the perfusion solution up to the region of the synapses. Additionally, in axons perfused with Ca²⁺-CaM there are regions where the synaptic gap between the membranes decreases from a control 4–6 to 2–3 nm. Both electrophysiological and morphological results can be interpreted as indicating that calcium-activated calmodulin acts directly on the junctional channels to induce their closure.     

The source of gibberellins in the parthenocarpic development of ovaries on topped pea plants

The role and source of gibberellins (GAs) involved in the development of parthenocarpic fruits of Pisum sativum L. has been investigated. Gibberellins applied to the leaf adjacent to an emasculated ovary induced parthenocarpic fruit development on intact plants. The application of gibberellic acid (GA₃) had to be done within 1 d of anthesis to be fully effective and the response was concentration-dependent. Gibberellin A₁ and GA₃ worked equally well and GA₂₀ was less efficient. [³H]Gibberellin A₁ applied to the leaf accumulated in the ovary and the accumulation was related to the growth response. These experiments show that GA applied to the leaf in high enough concentration is translocated to the ovary. Emasculated ovaries on decapitated pea plants develop without application of growth hormones. When [³H] GA₁ was applied to the leaf adjacent to the ovary a substantial amount of radioactivity accumulated in the growing shoot of intact plants. In decapitated plants, however, this radioactivity was mainly found in the ovary. There it caused growth proportional to the accumulation of CA₁. Application of LAB 150978, an inhibitor of GA biosynthesis, to decapitated plants inhibited parthenocarpic fruit development and this inhibition was counteracted by the application of GA₃ (either to the fruit, or the leaf adjacent to the ovary, or through the lower cut end of the stem). All evidence taken together supports the view that parthenocarpic pea fruit development on topped plants depends on the import of gibberellins or their precursors, probably from the vegetative aerial parts of the plant.Abbreviations FW flesh weight - GA_n gibberellin A_n - HPLC high-performance liquid chromatography     

Centrifugation of 2-cell mouse ova: cytoplasm stratification and recovery

Summary Two-cell mouse ova, which were centrifuged for l h at 70 000–90 000xg, showed a precise stratification of the cytoplasm and an elongation of the nucleus. The ova were fixed at different times and observed by light and electron microscopy using cytochemical methods and detergent extractions. Within 40 min after centrifugation the normal-looking morphology was recovered except for the persisting lipid caps at the centripetal poles of the blastomeres. Cleavage, compaction and blastulation were not prevented by centrifugation. Treatments with colcemid or cytochalasin D delayed but did not impair recovery. These results suggest that a resilient cytoskeletal structure may be involved in this kind of embryonic regulation.     

Microfungus-yeast mixed cultures in the degradation of amylaceous wastes. II: An experimental design for optimization of yeast production

Summary By means of an experimental factorial design the role of several culture conditions is described and its values determined for the maximum yeast production in mixed cultures ofAspergillus oryzae andRhodotorula glutinis.     

Ecology of microalgae in a high rate pond for piggery effluent purification in Singapore

Summary An integrated system for the collection, treatment and utilisation of piggery wastewater has been developed in Singapore which uses the cultivation of microalgae in high rate ponds to achieve reduction of BOD₅ and COD₅ of the effluent as well as producing single cell protein. A wide range of algal flora occurs in the ponds;Oocystis, Micratinium, Scenedesmus, Ankistrodesmus, Chlorella andOscillatoria spp were identified. Total algal counts, recorded from 1979 to 1981, ranged up to 10⁷ per ml of pond water. There were considerable variations in the algal population and in the predominating species. No discernible pattern was evident. Consequently pond operations were frequently disturbed by these fluctuations in population which in turn was attributed to the heterogeneous composition of the piggery waste, to variable weather conditions and to predation by larger organisms particularlyMoina. After passing through the ponds, the total suspended solids were removed by a novel dissolved air flotation method which gave a clear effluent showing an 87% reduction in BOD₅ value.

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Resumen En Singapur se ha desrrollado un sistema integrado para le recogida, tratamiento y utilización de aguas residuales de pocilgas. Este sistema usa el cultivo de microalgas en estanques, de caudal rápido afin de reducir las DBO₅ y DCO₅ del efluencte produciendo asimismo proteínas celulares. La flora de algas producida en estos estanques es amplia y variada, habiendose identificado:Oocystis, Micratinium, Scenesdemus, Ankistrodesmus, Chlorella y Oscillatoria. Los recuentos totales de algas tomados desde 1979 a 1981 llegaron a alcanzar 10;⁷ por ml de agua del estanque. Se observaron variaciones considerables tanto en la población total de algas como en las especies predominantes sin que se pudiese, determinar un patrón de variación característico. Estas fluctuaciones en la población, causantes de frecuentes alteraciones en el funcionamiento del estanque, se atribuyeron a la composición heterogenea de los resuduos, a las variaciones climáticas y la predación por otros organismos particularmenteMoina Después de su paso por los estanque los solidos suspendidos totales se eliminaron mediante un nuevo método de flotación con aire disuelto, obteniendose un efluente limpio con una reducción de la DBO₅ del 87%.

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Résumé Un système intégré pour la collecte, le traitement et l'utilisation d'eaux résiduaires de porcherie a été développé à Singapoure. Il utilise la culture de micro-algues tant pour la réduction accélérée en lagune de la DBO₅ et de la DCO de l'effluent que pour la production de protéines uni-cellulaires. On trouve une large gamme de flore algale dans les lagunes; des espèces d'-Oocystis, deMicratinium, deScenedesmus, Ankistrodesmus, deChlorella etd'Oscillatoria out été identifiées. Les comptages totaux d'algues, enregistrées de 1979 à 1981, ont donné jusqu' à 10⁷ cellules par ml d'eau de la lagune. On a observé des variations considérables de population algale tant quantitatives que qualitatives. On ne discernait pas de spectres évidents. En conséquence, les opérations lagunaires ont été fréquemment perturbées par ces fermentations en population, qui, à leur tour, ont été attribuées à la composition hétérogène de l'effluent de porcherie, aux conditions atmosphériques et climatiques variables et à la prédation par des organismes plus conséquents, plus particulièrement desMoina. Après passage par les lagunes, les solides totaux en suspension ont été enlevés par une méthode nouvelle de flottation à l'air dissous, qui a donné un effluent limpide, présentant une réduction de DBO₅ de 87%.

     

Active glycolysis and glycogenolysis in early stages of primary cultured hepatocytes. Role of AMP and fructose 2.6-bisphosphate

Summary This study examines the factors involved in the rapid glycolysis and glycogenolysis that occur during the first stages of hepatocyte culture: a) Shortly after seeding glycolysis, estimated as lactate released to culture medium, increased 10 times in comparison to that reported in vivo. By 8 to 9 h of culture, hepatocytes were nearly glycogen-depleted even in the presence of insulin. b) 6-Phosphofructo-2-kinase remained 100% active during this period. The proportion of the initial active phosphorylase (87%) decreased to 57% by 7 h of culture. c) Fructose 2,6-bisphosphate content was initially similar to that found in liver of fed animals, decreased after seeding and increased thereafter up to four times the initial concentration. In spite of changes in the concentration of this activator, the glycolytic rate remained high and constant. d) ADP and AMP increased sharply after cell plating, reaching values 1.7 and 3.5 times higher. The rise in AMP levels may be involved in the activation of glycolysis and glycogenolysis, because this metabolite is known to act as an allosteric activator of phosphofrucktokinase and glycogen phosphorylase. This metabolic situation resembles that of cells under hypoxia. Part of this work was presented at the 38th Annual Meeting of the Tissue Culture Association, Washington, DC, May 1987.     

Lesion and regeneration in the medial cerebral cortex of lizards.

The cerebral cortex of Squamate reptiles (lizards and snakes) may be regarded as an archicortex or "reptilian hippocampus". In lizards, one cortical area, the medial cortex, may be considered as a true "fascia dentata" on grounds of its anatomy, connectivity and cyto- chemo-architectonics of its main zinc-rich axonal projection. Moreover, its late ontogenesis and postnatal development support this view. In normal conditions, it shows delayed postnatal neurogenesis and growth during the lizard's life span. Remnant neuroblasts in the medial cortical ependyma of adult lizards seasonally proliferate. The late-produced immature neurocytes migrate to the medial cortex cell layer where they differentiate and give off zinc-containing axons directed to the rest of cortical areas. This results in a continuous growth of the medial cortex and its zinc-rich axonal projection. Perhaps the most important characteristic of the lizard medial cortex is that it can regenerate after having been almost completely destroyed. Recent experiments in our laboratory have shown that chemical lesion of its neurons (up to 95%) results in a cascade of events; first, those related with massive neuronal death and axonal-dendritic retraction and, secondly, those related with a triggered neuroblast proliferation and subsequent neo-histogenesis, and the regeneration of an almost new medial cortex that shows itself undistinguishable from a normal undamaged one. This is the only report to our knowledge that an amniote central nervous centre may regenerate by new neuron production and neo-histogenesis. Perhaps the medial cortex of lizards may be used as a model for neuronal regeneration and/or transplant experiments in mammals or even in primates.