Globule Leukocyte And Mast Cell in Bile Ducts of Cattle Naturally Infected with Liver Flukes

In n previous report dealing with the pathology of bovine fascio-liasis the author described an unknown cell type in the epithelium of bile ducts. The histological and histochemical investigations published in this paper suggest that the cell may be considered a globule leukocyte. Globule leukocytes are rare in uninfected livers but are occurring in abundance in main bile ducts of cattle with spontaneous fascioliasis and also in small perilobular ducts in dicrocoeliasis. Liver fluke infection causes an increase in the population of subepithelial mast cells. Mast cell and globule leukocyte present similarities In their cytochemical properties. However, at low pH toluldinc blue shows a stronger but Alcian blue a weaker affinity for mast cells than for globule leukocytes.     

Simultaneous Display of Multiple Foreign Peptides in the FliD Capping and FliC Filament Proteins of the Escherichia coli Flagellum

The bacterial flagellum is composed of more than 20 different proteins. The filament, which constitutes the major extracellular part of the flagellum, is built up of approximately 20,000 FliC molecules that assemble at the growing distal end of the filament. A capping structure composed of five FliD molecules located at the tip of the filament promotes polymerization of FliC. Lack of FliD leads to release of the subunits into the growth medium. We show here that FliD can be successfully used in bacterial surface display. We tested various insertion sites in the capping protein, and the optimal region for display was at the variable region in FliD. Deletion and/or insertion at other sites resulted in decreased formation of flagella. We further developed the technique into a multihybrid display system in which three foreign peptides are simultaneously expressed within the same flagellum, i.e., D repeats of FnBPA from Staphylococcus aureus at the tip and fragments of YadA from Yersinia enterocolitica as well as SlpA from Lactobacillus crispatus along the filament. This technology can have biotechnological applications, e.g., in simultaneous delivery of several effector molecules.     

Influence of 1-double bond and 11 beta-hydroxy group on stereospecific microbial reductions of 4-en-3-oxo-steroids

The yeast Rhodotorula glutinis and the anaerobic bacterium Clostridium paraputrificum were used for stereospecific reductions of 4-chloro-11 beta-hydroxy-17 alpha-methyl-testosterone and the corresponding 1-dehydro compound to prepare 5 alpha- and 5 beta-H derivatives, respectively. C. paraputrificum was able to 5 beta-reduce both substances, whereas the 5 alpha-reduction by R. glutinis was inhibited by the structure elements 1-en and 11 beta-OH so that the substrate with both structure elements was not 5 alpha-reduced. The microbial conversion of the four steroids with and without 1-en and 11 beta-OH was compared in semiquantitative experiments. A number of new substances are described, 11 beta-hydroxy and 11-oxo derivatives of 5 alpha- and 5 beta-dihydro-4-chloro-17 alpha-methyltestosterone including some 3-OH compounds and characterized by NMR, mass spectrometric and further data.     

Influence of a 9-double bond on stereospecific microbial 4,5-reductions

By stereospecific microbial reduction with Rhodosporidium rubrum or Rhodotorula glutinis, 17 alpha-cyano-methyl-4-estren-17 beta-ol-3-one was metabolized to 17 alpha-cyanomethyl-5 alpha-estrane-3 beta,17 beta-diol (50%) and 17 alpha-cyanomethyl-5 alpha-estrane-3 alpha,17 beta-diol (30%). By Clostridium paraputrificum the same substrate was reduced stereospecifically to 17 alpha-cyanomethyl-5 beta-estrane-3 alpha, 17 beta-diol (70%). When the corresponding 9-dehydrogenated compound 17 alpha-cyanomethyl-4,9-estradien-17 beta-ol-3-one (STS 557, a new progestagen) was fermented, yeasts failed in 5 alpha-reducing the 4-double bond. Still Clostridium paraputrificum formed the expected 5 beta-reduced metabolite 17 alpha-cyanomethyl-5 beta-estr-9-ene-3 alpha,17 beta-diol (60%). Structures were elucidated by n.m.r. and mass spectra and partly by circular dichroism. By oxidation of the metabolites, the corresponding 3-oxo compounds 17 alpha-cyanomethyl-5 alpha-estran-17 beta-ol-3-one, 17 alpha-cyanomethyl-5 beta-estran-17 beta-ol-3-one and 17 alpha-cyanomethyl-5 beta-estr-9-en-17 beta-ol-3-one were prepared. The evident influence of the 9-double bond on reduction of the 4-en-3-oxo compound STS 557 preventing 5 alpha-reduction but permitting 5 beta-reduction is discussed in view of the distinctly diminished metabolism of this progestagen in mammals.     

Farbstoffgehalt und Grösse der Augen bei verschiedenen Mutanten von Drosophila melanogaster

Zusammenfassung Quantitative Untersuchungen über den Farbstoffgehalt der Drosophilaaugen haben schon wiederholt gezeigt, daß die Werte bei bestimmten Mutanten von der Erwartung abweichen. So fand man regelmäßig bei den rotäugigen Mutanten v bzw. cn weniger Pterin und bei der braunäugigen Mutante bw weniger Ommochrom als bei Wildfliegen.Wir haben diese Befunde zunächst mit Hilfe einer vereinfachten Extraktions- und Meßtechnik nachgeprüft und bestätigt. Die genauere Analyse ergab dann aber, daß das Farbstoffdefizit der Mutanten v, cn und bw lediglich darauf beruht, daß diese Tiere kleinere Augen haben als die Wildfliegen. Die Augenverkleinerung ist jedoch nicht, wie gelegentlich vermutet wurde, die Folge einer polyphänen Wirkung der Gene v, cn und bw, sondern nur eine besondere Eigenschaft bestimmter Fliegenstämme, die heute in fast allen Laboratorien gehalten werden.Die Erscheinung selbst beruht auf der Wirkung augenverkleinernder Modifikationsgene, die bei diesen Stämmen zufällig mit den Farbgenen gekoppelt sind, durch geeignete Kreuzungen aber eliminiert werden können. Unsere so erhaltenen neuen v-, cn- und bw-Stämme besitzen nicht nur ebenso große Augen wie die Wildfliegen, sondern enthalten auch die theoretisch erwarteten Mengen an Augenfarbstoffen. Der Zusammenhang zwischen der Größe der Augen und ihrem Farbstoffgehalt hat u. a. zur Folge, daß die Männchen, die ja stets kleinere Augen haben als die Weibchen, bei allen Mutanten weniger Augenpigment besitzen als jene.Der Farbstoffgehalt der Augen hängt außerdem von der Zucht-temperatur ab. Fliegen, die sich bei 18° C entwickeln, besitzen weniger Pterin aber mehr Ommochrom als solche, die bei 26° C aufgezogen werden. Auch die Melaninsynthese im Integument der Tiere wird durch Temperaturerniedrigung begünstigt; aus 18°-Zuchten stammende Fliegen sind deutlich dunkler als die entsprechenden 26°-Tiere.     

Upper cretaceous rudist and stromatoporid associations of Central Oman (Arabian Peninsula)

Summary Rudist and stromatoporid associations of the Campanian from Central Oman are nearly monospecific. They are dominated byDurania aff.nicholasi, Vaccinites vesiculosus, Torreites milanovici or phaceloid and massive stromatoporids. Several other rudist genera play a secondary role. The thickness of the associations is rarely more than one metre. Solitary corals do not occur in the associations. Colonial corals are less common, although they are up to 1 m high and show considerable diversity. There are no binders. The reef structure indicates variable hydrodynamic conditions. They are always associated with very shallow water. The pureDurania aff.nicholasi patches with large colonial corals andTorreites milanovici are presumably the most rigid structures. The near monospecific associations ofVaccinites vesiculosus are widely distributed. Although mostly preserved in situ, strong currents, presumably caused by tropical storms, have repeatedly impaired and interrupted growth. The specific growth characteristics of the shell of some rudists, especially the radiolitids, enable an estimation of the individual lifespan. Frameworks of approximately 1 metre thickness probably developed in ±100 years. The sediments of the complete sections are predominantly bioclastic.     

Genetic recombination in mutants of the anthracycline-producer Streptomyces griseus

It was found that genetic recombination occurs if two marked strains of Streptomyces griseus (leukaemomycin-producing strains IMET JA 3933 and IMET JA 5142) are grown together in mixed cultures on semisolid media. The crossing techniques used and the method for carrying out selective analysis were essentially the same as those described by HOPWOOD (1967, 1972). The parent strains used for crosses were marked with single or double nutritional requirements and with mutations for drug resistance. The crosses are quite self-sterile, yielding only in one combination stable prototrophic recombinants at a low frequency (10(-5) to 10(-6)). The majority of recombinants behaved as stable haploid genotypes. A series of four-point crosses of different types of auxotrophs was carried out. The results of these experiments do not provide sufficient data for constructing a chromosome map, but provide basic information on the possibilities of genetic analysis of the production of anthracycline antibiotics. The majority of crosses performed were not fertile at 28 degrees C but, surprisingly, in some crosses carried out at 34 degrees C viable colonies were detected on minimal media at frequencies from 10(-3) to 10(-2).     

Flow cytogenetics of uncloned and cloned Chinese hamster cells

Flow cytometry has greatly facilitated the routine use of DNA content as a cellular indicator of the stages of the cell cycle and ploidy. DNA content can also be used to distinguish individual chromosomes. Fluorescent staining of chromosome DNA was done with a combination of ethidium bromide and mithramycin in hypotonic solution. Subsequent detergent treatment of the cells with Triton X-100 facilitated chromosome isolation. DNA flow cytometry of chromosomes of four established uncloned Chinese master cell lines showed 10 to 12 major subpopulations of chromosomes with varying degrees of overlap in the range of low and intermediate DNA content. Cloning of B14F28 cells, the line with the largest heterogeneity in chromosome number and DNA content, considerably reduced the dispersion in chromosome number and improved the resolution of DNA content distributions. Thus, cloned cells with a relatively homogeneous karyotype permit better discrimination of chromosome subpopulations by DNA content than uncloned cells and provide a more sensitive system to study mutagenic effects.