Saturated Fatty Acid Mutant of Saccharomyces cerevisiae with an Intact Fatty Acid Synthetase

To determine directly the effects of streptomycin on translational fidelity in intact cells, we studied the synthesis of beta-galactosidase and of the coat protein of bacteriophage R17 in an Escherichia coli mutant in which the bactericidal effects of streptomycin are delayed. After the addition of streptomycin to exponentially growing mutant cells, protein synthesis continues at an undiminished rate for approximately an hour; however, as measured by enzyme assays, little functional protein is produced. Serological assays designed to detect beta-galactosidase and bacteriophage R17 coat protein show that substantial amounts of the protein synthesized can react with antisera prepared against active beta-galactosidase and phage R17, indicating the aberrance of the protein produced in the presence of the antibiotic. The polypeptides synthesized in the presence of streptomycin are degraded in the cell to a much greater extent than protein synthesized in the absence of the antibiotic. The proteolytic attack on this protein is not affected by inhibitors of serine proteases, suggesting that enzymes other than those involved in "normal turnover" of cellular protein are responsible. In this strain, certain of the multiple effects of streptomycin are separated in time and the production of abnormal protein (enzymatically inactive and susceptible to proteolytic attack) could be studied in the absence of the lethal effect of the drug.     

The protein patterns of intracytoplasmic membranes and reaction center particles isolated fromRhodopseudomonas capsulata

Intracytoplasmic membranes of wild type strain 37 b 4 and mutant strains A1a car^-bchl^-, A1a car^-bchl⁺ ofRhodopseudomonas capsulata were isolated. The membrane proteins were solubilized and separated by polyacrylamide gel electrophoresis (methods of Takayamaet al., 1964; Weber and Osborn, 1969). The band patterns were compared with each other. From the strain A1a car^-bchl⁺ reaction center particles were isolated by treatment of membrane with Triton X-100 followed by sucrose density gradient centrifugation. The reaction center particles were found to be enriched in reaction center bacteriochlorophyll. This pigment shows a reversible bleaching at 855 nm and a blue shift at 798 nm. The light harvesting bacteriochlorophyll portion of this fraction was 14–22% of the total bacteriochlorophyll content. The three main proteins of the reaction center particles amount to about 80% of the total protein of the particles. The molecular weights of the main proteins were estimated to be 32000, 27500 and 22500 daltons.     

Serum-induced inhibition of isotonic fluid absorption by the kidney proximal tubule I. Mechanism of inhibition

Isotonic reabsorption by the rat kidney proximal tubule was drastically inhibited after less than 2 min intraluminal perfusion with fresh sera from rat (both homologous and autologous), cat, rabbit and human, but not with sera from mouse and guinea pig. The inhibitory factor in serum in a heat (56° C for 30 min) and storage (4°C for 2–5 days) labile macromolecule (mol. wt 50 000) and requires Ca²⁺ for its effect. The cellular electrical potential difference of the proximal tubular cells was irreversively destroyed and intraluminally perfused trypan blue dye incorporated into the tubular cells after the intraluminal perfusion with serum for 2 min. These observations suggest that lysis of the proximal tubular cells is the mechanism for serum-induced inhibition of proximal tubular isotonic reabsorption.     

Morphologische Untersuchungen an der Glandula bulbourethralis der Katze

Zusammenfassung Das Parenchym der Glandula bulbourethralis der Katze besteht aus weitlumigen, gebuchteten intraglandulären Gängen, in welche kurze, englumige, zumeist unverzweigte Tubuli einmünden. Der Drüse fehlt eine äußere Organkapsel, so daß ihre peripheren Tubuli stellenweise direkt zwischen den Fasern des quergestreiften M. bulboglandularis liegen. Die Drüsentubuli und die Buchten der intraglandulären Gänge sind mit einem einschichtigen Zylinderepithel ausgekleidet, auf den Gangfalten ist das Epithel abschnittsweise mehrreihig, Die sezernierende Epitheloberfläche ist durch die Ausbildung von interzellulären Sekretkapillaren vergrößert. Breite Zwischenzellspalten (Durchmesser etwa 1,5), in welche schlanke interdigitierende Cytoplasmafortsätze hineinragen, erstrecken sich von der Basalmembran bis kurz unter das Tubulusbzw. Ganglumen. Die lumenseitigen Zellgrenzen tragen einige stummelförmige Mikrovilli und besitzen zerklüftete Außenkonturen, die durch glykogenreiche Cytoplasmaprojektionen bedingt sind. Alle Epithelzellen sind reich an Mitochondrien. Die supranuklearen Abschnitte der meisten Gang- und Tubuluszellen enthalten Sekretgranula, welche im Elektronenmikroskop unterschiedliche optische Dichten aufweisen können. Die Granula enthalten ein PAS-positives, neuraminsäurehaltiges epitheliales Muzin, das in einzelnen Sekretkörnchen auch eine histochemische Reaktion auf Sulfatgruppen gibt. Alle Epithelzellen reagieren sehr stark auf unspezifische Esterase und stark auf -D-Glucuronidase, -D-Glactosidase sowie die Enzyme des Citronensäurezyklus, der Glykolyse und der Atmungskette (NAD-ICDH, SDH, ALD, LDH, ADH, GDH, NADH-T-Red, Cyt-Ox).

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Morphological studies on the bulbourethral gland of the male cat

Summary The bulbourethral glands of sexually mature male cats are studied with the light and electron microscope. The parenchyma consists of spacious, sinus-like intraglandular ducts and short, narrow, mostly unbranched tubular endpieces. The gland has no complete connective tissue capsule, consequently some of the peripheral tubules are situated directly in between the fibers of the surrounding bulboglandularis muscle. The endpieces and the sinus of the intraglandular ducts are lined by a simple columnar epithelium, whereas the folds of the ducts are generally covered by a low pseudostratified epithelium. The secretory surface of the cells is increased by intercellular canaliculi which communicate with the gland lumen. These canaliculi are identified on the light microscopic level by their strong 5-nucleotidase activity. Furthermore widened intercellular spaces (approximately 1,5 in diameter) filled with slender, interdigitating cytoplasmic processes extend from the basal lamina to the apical junctional complexes. The luminal cell pole exhibits some short microvilli and forms irregularly shaped, glycogen containing protrusions. Within the cytoplasm of the gland cells numerous spherical mitochondria, some dense bodies, a typical Golgi apparatus, free ribosomes and a poorly developed endoplasmic reticulum are to be observed. Secretory granules which can be grouped into three types on the basis of their electron density occur in the supranuclear regions of most of the cells. According to histochemical tests all granules contain a periodate reactive sialomucin and some of them also sulfate groups. The glandular parenchyma is site of an exceptionally strong unspecific esterase activity and is rich in -D-glucuronidase, -D-glactosidase, aldolase, -glycerophosphate dehydrogenase, lactate dehydrogenase, alcohol dehydrogenase, NAD-dependent isocitrate dehydrogenase, succinate dehydrogenase and cytochrome oxydase.

Mit dankenswerter Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Der cis-trans-Positionseffekt in Heterokaryen morphologischer Mutanten von Podospora anserina

Zusammenfassung Der cis-trans-Positionseffekt in Heterokaryen der nicht gekoppelten morphologischen Mutanten z und oct-1 von Podospora anserina wird durch quantitative Kernunterschiede vorgetäuscht.Im cis-Fall führt die Wuchsverminderung von Hyphen mit überwiegendem Gehalt an Doppelmutantenkernen nach Mikrosektorenbildung im Mycel zu einer Selektion der Hyphen mit überwiegendem Wildkerngehalt. Die Doppelmutantenkerne sind nach 6 cm Wuchsstrecke aus dem Mycel verschwunden. Dieses weist daher Wildphänotyp auf.Im trans-Fall wird eine Kernentmischung durch Anastomosenbildung kompensiert, da die entestehenden Kleinsektoren gleich schnell wachsen. Das Mycel bringt daher die oct-1- und z-Merkmale in einem intermediären Phänotyp zum Ausdruck.

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The cis-trans position effect in heterocaryons of morphological mutants of Podospora anserina

Summary The cis-trans position effect of the two non-linked morphological mutants z and oct-1 in heterocaryons of Podospora anserina is caused by quantitative differences of the two types of nuclei.In the cis configuration, there is a growth diminuition of hyphae containing predominantly nuclei of the double mutant type which leads to a selection of hyphae containing mainly the wildtype nuclei. This is due to the formation of microsectors. After 6 cm of mycelial growth the double mutant nuclei are no longer distinguishable; the mycelium adopts the wild phenotype.In the trans configuration, the microsectors of both types of mutant nuclei exhibit the same growth rate; the separation of nuclei is prevented as a result of the formation of hyphal anastomoses. The mycelium shows then an intermediate phenotype with both mutant characteristics.

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Mit Unterstützung der Deutschen Forschungsgemeinschaft.     

Chromosome Replication in Salmonella typhimurium

The replication of the Salmonella typhimurium chromosome was studied. As with E. coli 15T(-), replication was sequential. After amino acid starvation, replication proceeded from a unique and heritable region of the chromosome. 5-Bromouracil, when substituted for thymine, did not disturb the sequence of replication nor did it initiate extra replication cycles. By labeling the origin and the terminus of the chromosome with (3)H- and (14)C-thymine, respectively, it was possible to determine that the rate of chain elongation decreases as the growth rate decreases. No gap in the replication cycle could be observed.     

Zur Morphologie und Phylogenie der Schizorhynchia (Turbellaria,Kalyptorhynchia)

The proboscis organs of 21 species of Schizorhynchia represent new characters. The meaning of apomorph characters of the proboscis organ and of certain parts of the genital apparatus is discussed. In the phylogenetic system the position of the four families of Schizorhynchia is considered.

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Gefördert durch eine Sachbeihilfe der Deutschen Forschungsgemeinschaft im Schwerpunktprogramm Litoralforschung — Abwassereinflüsse in Küstennahe.     

Feinstrukturen der Mikrokörper (Microbodies) des proximalen Nierentubulus

Zusammenfassung Die Feinstrukturen der Mikrokörper des Nierenepithels werden beschrieben und mit denjenigen der Leber-Mikrokörper verglichen. Als besondere Charakteristika der Nieren-Mikrokörper sind eine (nicht kristalline) nucleoide Verdichtung und eigentümliche stabförmige Ausstülpungen (Stäbe) anzusehen. Die Stäbe stellen unterschiedlich lange Zylinder mit einem Durchmesser von 100 nm dar. Im Inneren findet sich eine unmittelbar der Membran anliegende, ring- oder spiralförmig angeordnete, granuläre Struktur (Granula-Durchmesser 50 Å), die in Stablängsschnitten eine Querstreifung vortäuscht. Es wird eine in Phasen ablaufende Bildung der Mikrokörper-Stäbe angenommen: ein in der Matrix entstandener Granula-Zylinder hebt sich aus dem Mikrokörper heraus, wobei die Mikrokörper-Membran entsprechend vorgebuchtet wird, und wächst schließlich zu einem eigenständigen, allseits membranumzogenen Stab aus. Die Möglichkeit, daß die Stäbe von Mikrokörpern abgestoßen werden, wird diskutiert. — Die Segregation von Mikrokörpern in Vakuolen wird nicht als aktive Beteiligung an lytischen Prozessen, sondern als autophagischer Vorgang gedeutet.

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Fine structure of microbodies in proximal tubular epithelium of the kidney

Summary Ultrastructural observations on microbodies in normal proximal tubule cells of the rat kidney are described and compared with microbodies of hepatic parenchymal cells. After fixation in osmium tetroxide with phosphate buffer the special features of the renal microbodies are the non-crystalline nucleoid and protrusions (rods) extending from the main body. These rods are cylindrical in shape having a diameter about 100 nm and are of varying lengths. Inside the limiting membrane are ring- or spiral-like ordered profiles consisting of granules (about 50 Å in diameter) which often appeared as a row of parallel linear densities arranged at approximately right angles to the long axis of the rod. It can be demonstrated that the parallel linear pattern depends on the projection of the granules in the photographic plane. — The findings suggest that the cylindrical structures of granules are formed in the peripheral matrix of microbodies; in a second phase they are lifted outside, in part enveloped with the membrane of the microbody; in this situation, the protrusions are formed. This form of creation would explain the characteristic excentrical (tangential) relation between protrusions and the main body. The observation that rods are often seen apparently isolated in the cytoplasm without visible connection with a microbody is only discussed hypothetically, because of the plane of sectioning. — Microbodies and rods can be identified in cytosegresomes. These investigations were interpreted as an autophagic degradation and not as an active role of the enzymes of microbodies in digestive mechanisms.

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Herrn Prof. Dr. Helmut Ruska zum 60. Geburtstag gewidmet.

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.

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Herrn Doz. Dr. W. Thoenes danke ich für wertvolle Hinweise und für die kritische Durchsicht des Manuskriptes.     

Zum Bewegungssehen des Mehlkäfers Tenebrio molitor

Summary The locomotory turns of the beetle Tenebrio molitor were recorded from free walking animals which were held in stationary position and orientation by means of a tread compensator. Striped patterns revolved around an animal elicit optomotor responses which undergo an inversion at smaller stripe widths. The position of the inversion point characterizes the resolving power of the motion detectors, and corresponds to a 6.5° average angle between the optical axes of interacting elements in the motion detecting device. This indirectly determined value is consistent with the anatomically measured 7° average angle between the axes of neighbouring ommatidia in the Tenebrio compound eye.