Different effects of the glucosidase inhibitors 1-deoxynojirimycin, N-methyl-1-deoxynojirimycin and castanospermine on the glycosylation of rat alpha 1-proteinase inhibitor and alpha 1-acid glycoprotein.

The glucosidase inhibitors 1-deoxynojirimycin, N-methyl-1-deoxynojirimycin and castanospermine were used to inhibit oligosaccharide processing in primary cultures of rat hepatocytes. Their effect on the glycosylation of alpha 1-proteinase inhibitor (alpha 1PI) and alpha 1-acid glycoprotein (alpha 1AGP) was studied. Of the three glucosidase inhibitors examined, 1-deoxynojirimycin inhibited not only oligosaccharide trimming but also glycosylation de novo of newly synthesized proteins, resulting in the formation of alpha 1PI with two and three (normally carrying three) and alpha 1AGP with two to five (normally carrying six) oligosaccharide side chains. In the presence of the glucosidase inhibitors, glucosylated high-mannose-type oligosaccharides accumulated. Whereas most of the endoglucosaminidase-H-sensitive oligosaccharides formed in the presence of 1-deoxynojirimycin contained only one glucose residue, N-methyl-1-deoxynojirimycin and castanospermine led mainly to the formation of oligosaccharides with three glucose residues. None of the three glucosidase inhibitors completely prevented the formation of complex-type oligosaccharides. Thus, in their presence, alpha 1PI and alpha 1AGP with a mixture of both high-mannose and complex-type oligosaccharides were secreted.     

In vivo visualization of individual neurons in arthropod ganglia facilitates intracellular neuropil recording

Summary A simple method for the in vivo visualization of dye filled cells by laser illumination is used to characterize neurons in situ in the segmentai ganglia of the locust and the crayfish (Fig. 1). Neuron visualization provides the structural information necessary for identification of cells during an ongoing physiological experiment (Figs. 2, 3). Sequential penetrations of soma and neuropil as well as simultaneous double neuropil penetrations of spiking and nonspiking cells are facilitated by the visual control afforded by neuron visualization (Figs. 4, 5, 6). Furthermore, neuron visualization allows the sampling of cellular properties at multiple, predetermined sites in the dendritic and axonal arbors of identified neurons (Fig. 7) and aids in establishing synaptic connectivity through double neuropil recordings (Fig. 8).     

THE SEPARATION OF DIFFERENT CELL CLASSES FROM LYMPHOID ORGANS : III. The Purification of Erythroid Cells by pH-Induced Density Changes

1. Mammalian erythrocytes swell as the pH of the isotonic suspending medium is lowered, as a direct consequence of the specialized permeability properties of the erythrocyte membrane. Lymphocytes and granulocytes from a variety of sources did not exhibit this property. 2. The behaviour of mouse bone marrow erythroid cells at various stages of differentiation was studied by using a change in buoyant density with pH as an index of swelling. The ability to swell with a pH drop was acquired while the cell was still nucleated. All non-nucleated cells showed swelling. Most small erythroblasts shared this property, whereas most large erythroblasts did not. 3. The density shift with pH was used to provide a purification scheme specific for erythroid cells. The bone marrow cells were first centrifuged to equilibrium in an isotonic albumin density gradient at neutral pH. Regions of the gradient containing the erythroid cells were collected, and the cells were recovered and redistributed in an albumin gradient at acid pH. The erythroid cells showed a specific density shift which removed them from contaminants. Preparations containing 90–97% erythroblasts were obtained by this technique. 4. Differentiation within the erythroid series was accompanied by a general increase in cell buoyant density at neutral pH. This density increase may have been a discontinuous process, since erythroid cells appeared to form a number of density peaks. 5. The pH shift technique, in association with established density distribution and sedimentation velocity procedures, provides a range of cell separation techniques for biological or biochemical studies of erythroid cell differentiation in the complex cell mixtures in bone marrow or spleen.     

Light-dependent hydrogen evolution by Scenedesmus

Summary The effect of glucose and the uncoupler Cl-CCP upon hydrogen production was studied in adapted cells of Scenedesmus obliquus D₃. Cl-CCP at 10^-5M concentration completely inhibited the evolution of H₂ in the dark and increased the apparent rate of H₂ evolution in the light. At 10^-5M Cl-CCP, photosynthesis and photoreduction by anaerobically adapted algae were only temporarily inhibited; O₂ evolution reappeared after approximately 1 hr of illumination if CO₂ was present. Increasing the Cl-CCP concentration to 5 x 10^-5M led to a maximum rate of photohydrogen production and fully inhibited H₂ evolution, photoreduction and dark H₂ evolution. H₂ evolution was accompanied by a release of varying amounts of CO₂ in the light, as well as in the dark. Dark CO₂ production was stimulated by Cl-CCP. H₂ evolution in the light was stimulated by adding glucose to autotrophically grown cells or by growing the cells heterotrophically with glucose; starvation had an opposite effect. Adapted cells released ¹⁴CO₂ from the 3 and/or 4 position of specifically labeled glucose, indicating that degradation occurred via the Embden-Meyerhof pathway. The amount of H₂ released by autotrophically grown cells was the same either with continuous illumination or with short periods of light, followed by darkness. Scenedesmus mutant No. 11, which is unable to evolve O₂ was not inhibited in its capacity to evolve H₂ in the light. These data indicate that the evolution of H₂ in the light by adapted Scenedesmus depends upon the degradation of organic material and does not require the production of free O₂ by photosystem II.The following abbreviations are used: Cl-CCP = carbonyl cyanide m-chlorophenylhydrazone; DCMU = 3-(3,4-dichlorophenyl)-1,1-dimethylurea, DNP = 2,4-dinitrophenol.This work was supported by contract AT-(40-1)-2687 from the U.S. Atomic Energy Commission.     

Zur Feinstruktur der Blutmonozyten des Haushuhns

Zusammenfassung Blutmonozyten von 10 klinisch gesunden, legenden weißen Hybridhennen wurden elektronenmikroskopisch untersucht. Dabei wurde eine zweite Fixierungsmethode in Anlehnung anHirsch u.Fedorko (1968) angewandt. — Deutlich ausgeprägte Golgi-, ER-sowie vesikuläre bzw. granuläre Strukturen kennzeichnen die Aktivität der Monozyten. Sie enthalten außerdem feine Filamente, die stellenweise durch Querbrücken verbunden sind, Mikrotubuli sowie in einem Fall phagozytierte zelluläre Partikel.

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Fine structure of the hen's monocytes

Summary The fine structure of the monocytes from 10 clinically healthy hybrid hens was investigated electron microscopically. A second fixation method afterHirsch andFedorko (1968) was used. — The activity of the monocytes is characterised by a well developed Golgi apparatus, ER- and vesicular respectively granular structures. Furthermore the cells contain fine filaments with cross-bridges, microtubules and - in a single case-phagocytized cellular particles.

     

Zur Verbreitung der Mallophagen der Gattung Myrsidea Waterston auf der Dschungelkrähe Corvus macrorhynchos Wagler

Im allgemeinen ist eine Mallophagengattung auf eine einzige Familie oder Ordnung ihrer Wirte beschränkt; seit N itzsch (1815) und K ellogg (1896) konnte diese Erscheinung immer wieder bestätigt werden. Damit ist auch naturgemäß die Verbreitung der Mallophagen identisch mit der ihrer Wirte; allerdings können extreme Klimate (s. E ichler , 1963, und N iethammer , 1962) das Vorkommen der Mallophagen im Gegensatz zu dem ihrer Wirte beschränken. Trotz dieser Ausnahmen ist es jedoch wegen des engen Wirt-Parasit-Verhältnis grundsätzlich möglich, mit Hilfe der Mallophagen-Verbreitung den Wirt betreffende, taxonomische, phylogenetische und zoogeographische Fragen zu erörtern. Wie erfolgreich diese Untersuchungsmethoden sind, zeigen etwa die Arbeiten von C lay (1961, 1964, 1966 a, b), E ichler (1963), E lbel & E merson (1959) und T immermann (1957, 1965).     

Induktive Bildung partikelgebundener Uricase bei Hydrogenomonas H16 und anderen aeroben Bakterien

Zusammenfassung Induktive Bildung von Uricase (E.C. 1.7.3.3. Urat: O₂-Oxidoreductase) wurde während des Wachstums mit Harnsäure als alleiniger Kohlenstoff-und Stickstoffquelle bei Hydrogenomonas H 16, Micrococcus denitrificans und Pseudomonas aeruginosa beobachtet.Das Enzym wurde in der Partikelfraktion nachgewiesen, die aus Ultraschallextrakten bei 100 000 g sedimentiert.Gewaschene Partikeln aus Hydrogenomonas H 16 verbrauchten 0,45 Mole O₂ je Mol Harnsäure, gemessen in Boratpuffer beim Optimum von pH 9,0. Die Reaktion ist empfindlich gegenüber Cyanid; 4 · 10^-6 m KCN führt zu einer 50%igen Hemmung.

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Inductive biosynthesis of particle-bound uricase in Hydrogenomonas H 16 and other aerobic bacteria

Summary Induced biosynthesis of uricase (E.C. 1.7.3.3 Urate: O₂ oxidoreductase) was observed during growth with uric acid as the only carbon and nitrogen source in strains of Hydrogenomonas H 16, Micrococcus denitrificans and Pseudomonas aeruginosa.The enzyme was located in particles, sedimenting from ultrasonic preparations at 100000 g.An average of 0.45 moles of oxygen were utilized during the oxidation of one mole of uric acid by washed particles from Hydrogenomonas H 16 in borate buffer at the pH optimum of 9.0. The reaction is sensitive to cyanide; 4 · 10^-6 m KCN caused a 50% inhibition.