Host Cell Deoxyribonucleic Acid Polymerase I and the Support of T4 Bacteriophage Growth

Ultraviolet irradiation of Escherichia coli polA(-) cells reduces their capacity to support the growth of T4 phage. There is no additional loss of capacity observed in pol tsA(-)recA(-) double mutants at the nonpermissive temperature. The reversion frequency of a T4 rII mutant after ultraviolet irradiation is not changed by the absence of host deoxyribonucleic acid polymerase I.     

Enterochelin System of Iron Transport in Escherichia coli: Mutations Affecting Ferric-Enterochelin Esterase

Three mutant strains of Escherichia coli have been isolated which are lacking ferric-enterochelin esterase activity. This enzyme catalyzes the hydrolysis of the enterochelin moiety of ferric-enterochelin to yield ultimately three molecules of N-2,3-dihydroxybenzoylserine. The mutants (designated fes(-)) were shown to be unaffected in enterochelin biosynthesis, capable of enterochelin-mediated iron uptake, and able to utilize ferric-dihydroxybenzoylserine complexes normally. When grown under iron-deficient conditions, however, they showed an absolute requirement for added iron or citrate, a phenotype characteristic of mutants defective in some part of the enterochelin system of iron uptake. These results support the theory that iron, taken up by the cell as ferric-enterochelin is only available for general cell metabolism after hydrolysis of the ligand by enterochelin esterase. The three fes(-) strains were shown to be affected in the B component of enterochelin esterase. The fesB gene which is probably the structural gene coding for component B of the esterase, was shown to be located at about minute 14 on the E. coli chromosome together with seven other genes involved in the enterochelin system of iron transport.     

Ecto-ganglioside-sialidase activity of herpes simplex virus-transformed hamster embryo fibroblasts

Cellular location of ganglioside-sialidase activity was determined in confluent hamster embryo fibroblasts transformed with herpes simplex virus type 2. Approximately equal specific activities of ganglioside-sialidase activity were found to be associated with the crude lysosomal and crude plasma membrane fractions isolated from whole cell homogenates. Whole transformed cells hydrolyzed exogenous ganglioside substrate, suggesting a partial location of the cellular sialidase on the outer surface of the plasma membrane of these cells. Intact cells were treated with the diazonium salt of sulfanilic acid, a nonpenetrating reagent inhibitory to ecto-enzymes (DePierre, J.W., and M. L. Karnovsky. 1974. J. Biol. Chem. 249:7111-7120). Cytoplasmic lactate dehydrogenase activity was not inhibited by this treatment, and mitochondrial succinate dehydrogenase activity was inhibited only 10%, indicating that intracellular enzymes were not affected. 5'-Nucleotidase activity was diminished 90%, and sialidase very rapidly lost 40% of its exogenously directed activity. These results show that, in herpes simplex virus-transformed fibroblasts, ganglioside-sialidase is both a lysosomal and a plasma membrane enzyme. The plasma membrane sialidase is capable of acting on endogenous plasma membrane sialolipids and also functions in the cultured transformed cell as an ecto-enzyme which can attack exogenous substrates.     

Feinstruktur der Leberzellen vonOxidus gracilis (C.L. Koch, 1847) (Diplopoda,Paradoxosomatidae)

Zusammenfassung Bei den Vertretern einiger Diplopoden-Familien ist der Mitteldarm von einer zelligen Hüllschicht umgeben. Die Perikaryen stehen dicht bei dicht, basale Ausläufer durchziehen die Fasern der Muskularis und verzweigen sich sekundär beim Eintritt in die sehr dicke Basallamina des Mitteldarmepithels. Die terminalen Endigungen jeder Zelle dringen in ein extrazelluläres basales Labyrinth je einer ausdifferenzierten Mitteldarmzelle ein und verflechten so die Hüllschicht mit dem Darmepithel. Darüberhinaus scheint es Fusome zwischen den Endaufzweigungen und der Mitteldarmzelle und damit die Möglichkeit eines intrazellulären Stofftransports zwischen beiden Zellen zu geben.Die Hüllschicht ist kein Epithel, denn Zellkontakte zwischen benachbarten Zellen fehlen ebenso wie eine gemeinsame Basallamina. jede Zelle ist von ihrer eigenen, dünnen Basallamina umgeben. Sie wird außerdem bei normal ernährten Individuen durch einen großen Vorrat an Glykogen und zahlreiche Zytosomen charakterisiert. Viele Mitochondrien sind dadurch vor allem in die Zellperipherie oder in die basalen Ausläufer gedrängt. In deren Nähe findet man auch überwiegend glattes endoplasmatisches Retikulum, und auch freie Ribosomen sind nicht selten. Der Zellkern vermittelt den Eindruck hoher Stoffwechselaktivität; ein Nukleolus ist immer vorhanden. Mikrotubuli sind im Zellkortex parallel zur Oberfläche orientiert, vor allem aber in den basalen Ausläufern in Längsrichtung. Obwohl kein Fett gespeichert wird, kann man die Zellen mit dem Chloragog-Gewebe der Anneliden oder mit den Leberzellen der Vertebraten vergleichen. Sie scheinen von den Mitteldarmzellen Nährstoffe zu übernehmen, diese zu verarbeiten, die Produkte freizusetzen oder zu speichern und dabei wertlose stickstoffhaltige Moleküle in den Zytosomen abzulagern. Der gebräuchliche Terminus Leberzellen ist daher angebracht.Tiere, die 14 Tage ohne Nahrung gehalten wurden, zeigen eine erhebliche Schrumpfung der Leberzellen. Deren Plasmalemm ist nur an einigen Stellen mit der Basallamina verbunden, überall sonst ist der extrazelluläre Raum zwischen Zellmembran und Basallamina stark erweitert. Das Glykogen ist größtenteils abgebaut. Der elektronenoptische Kontrast der verschiedenen Zellelemente ist äußerst schwach. Die Mikrotubuli sind völlig verschwunden. Nur die Zytosomen heben sich weiterhin deutlich ab. Aber auch sie scheinen umgewandelt und zu wenigen Ansammlungen zusammengeflossen zu sein. Dies deutet auf Mobilisierung des gespeicherten Glykogens hin, kann aber auch beginnende Degeneration der Leberzellen signalisieren.

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Fine structure of the liver cells ofOxidus gracilis (C.L. Koch, 1847) (Diplopoda, Paradoxosomatidae)

Summary In some families of Diplopods the midgut is surrounded by an enveloping cell layer. Basal runners of their pericarya get through the longitudinal and transversal muscle fibres and ramify secundarily before entering the very thick basement membrane of the midgut epithelium. The terminal branches of each cell enter into a basal labyrinth of only one differentiated midgut cell interlacing intensively with the epithelium in this way. Moreover it seems that there are open junctions (fursomes) between terminal branches and the midgut cell and therefore an intracellular transport of materials.The enveloping layer is no epithelium. THere are no typical contacts between neighbouring cells; each enveloping cell is surrounded by its own thin basement membrane, a common basement membrane does not exist. Enveloping cells of normally nourished individuals are characterized by a large storage of glycogene in the cytoplasm. Besides this there are many distinct cytosomes which point at a significant activity in metabolism. Many mitochondria, which mostly pushed off towards the cell periphery or lie in the primarily ramifications indicate the same. Close by there is also prevailing smooth endoplasmic reticulum. Free ribosomes are not rare. The nucleus gives the impression of activity in metabolism too; a nucleolus does exist. Microtubules arranged parallely to the cell surface in the cortex, but especially in the basal ramifications in longitudinal direction can be seen. Though lipoids are not being stored, one might compare these cells with the chloragog tissue in annelids or with the liver cells in vertebrates. They seem to accept food from the midgut cells, to digest and release, respectively store it temporarily, depositing at that valueless nitrogene containing molecules into the cytosomes. The usual term liver cell therefore is wellchosen.Animals after 14 days without food signify a large shrinkage of the liver cells. Plasmalemma is fixed only at certain points to the basement membrane; the extracellular space between them is widened extremely. Glycogene mostly is removed. The electron microscopically visible contrast of the cell elements is very faint. Microtubules have disappeared. Cytosomes, however, contrast distinctly. But they, too, seem to be transformed and joined into a few accumulations. This points to mobilization of the stored glycogene, but in the same manner also to symptomes of degeneration brought into action.

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Herrn Professor Dr. Dr. h.c. W.E. Ankel zu seinem 80. Geburtstag am 7.8.1977 gewidmet     

Endocytosis in Chang liver cells. Quantitation by sucrose- 3 H uptake and inhibition by cytochalasin B

The addition of 0.08 M sucrose to a culture medium containing Chang-strain human liver cells causes intense cytoplasmic vacuolation. Electron microscopy of these cells grown inferritin, time-lapse cinematography, and radioautography reveal that the vacuoles arise by endocytosis and that the sucrose is taken into the cell and localized in the vacuoles. Tracer studies demonstrate that sucrose-³H provides a marker for quantitation of endocytosis and that it neither induces nor stimulates endocytosis. Electron micrographs of vacuolated liver cells show microfilaments in close proximity to the inside of the plasma membrane, in the pseudopodia, and to the cytoplasmic side of the membrane surrounding endocytosis vacuoles. Cytochalasin B (CB), a mold metabolite that inhibits various types of cell motility, has a dose-dependent inhibitory effect on the uptake of sucrose-³H by these cells. This inhibition is accompanied by a cessation of the movement of ruffles and pseudopodia on the surface of the cells and the formation of blebs which arise from the cell''s surface. These morphological changes are quickly reversible upon removal of CB. Alterations in the appearance and location of microfilaments are also observed in CB-treated cells.     

Rapid prenatal and postnatal detection of inborn errors of propionate,methylmalonate, and cobalamin metabolism: A sensitive assay using cultured cells

Summary A sensitive, reliable, and easily performed procedure is described for the prenatal and postnatal detection of inborn errors of propionate, methylmalonate, and cobalamin metabolism using cultured amniotic cells and skin fibroblasts. With this assay, control fibroblast lines incorporated a mean of 6.89 nanoatoms ¹⁴C/mg protein from [1-¹⁴C]propionate into trichloroacetic acid (TCA)-precipitable cell material in 10h. Twenty-five mutant fibroblast lines from patients with propionicacidemia or one of the methylmalonicacidemias fixed 0.04 to 0.93 nanoatoms ¹⁴C/mg. Considerable variation was observed, both among and within discrete mutant classes, with respect to the residual amount of propionate pathway activity, possibly reflecting further molecular heterogeneity in these disorders.We applied this procedure to cultured amniotic cells from controls and 4 midtrimester pregnancies at risk for methylmalonicacidemia and diagnosed one fetus with a methylmalonyl CoA apomutase defect and 3 fetuses which were unaffected.Presented in part at the annual meeting of the Society for Pediatric Research, St. Louis, Missouri, April 1976.     

Prednisone in MOPP chemotherapy for Hodgkin's disease.

High remission rates have been produced by MOPP (mustine, vincristine, procarbazine, and prednisone) chemotherapy in patients with advanced Hodgkin''s disease, but the prednisone component has caused adverse effects in patients who have undergone radiotherapy. The remission rates and length of remission were reviewed in 211 patients with Hodgkin''s disease who received chemotherapy either with or without prednisone. In contrast to the findings of a British study, there were no significant differences in remission rates or length of remission between patients who had received prednisone and patients who had not. There were differences between the British prospective study and this retrospective one, but it is difficult to know what accounted for the substantial differences in the findings.