Betrachtung über den Energiekonsum von Tieren mit Atmungsorganen von zweierlei Typ

Zusammenfassung Besitzt eine isometrisch wachsende Tierart zweierlei Atmungsorgane, von denen das eine O₂ prop. L ² aufnimmt und das andere prop. L ³, so ergibt sich in einem Koordinatensystem mit log L als Abszisse und log (O₂) als Ordinate für die Abhängigkeit des (O₂) von L keine Gerade, sondern eine gegen die Ordinate konkave, Linie (Abb. 1), die bei L=O mit der Steigung a=tg =2 beginnt, für L=1 den Wert 2,5 erreicht und bei L 8 mit der Steigung 3 endet. (Bei log G als Abszisse betragen die entsprechenden Werte 2/3, 5/6 und 1.) Für Längenintervalle von einer Zehnerpotenz (also Gewichtsintervallen von 10³) dürfte im Rahmen der experimentellen Breiten oft eine anscheinend lineare Abhängigkeit gefunden werden. Ergibt sich dann empirisch für ein Längenintervall l10 (mit der mittleren Länge L _m) oder das entsprechende Gewichtsintervall l10³ eine Atmungsgerade mit der Steigung , so kann unter den obigen Voraussetzungen vermutet werden, daß sich die durch beide Organe konsumierten O₂-Mengen verhalten wie (3 — )( — 2). Bei der Körperlänge L * (Formel 9), die nicht realisiert zu sein braucht, müßten beide Organe gleichviel O₂ aufnehmen. Die Anwendung dieser Folgerungen auf Tiere, die möglicherweise Atmungsorgane von mehrererlei Typ besitzen (Insekten, Insektenlarven, Isopoden, Schnecken), sowie die Erörterung solcher empirischer Atmungsgeraden, die eine zwischen 2 und 3 liegende Steigung a aufweisen, wird in den weiteren Mitteilungen erfolgen.Zugleich Mitt. IX der Reihe Körpergröße, Körperzeiten und Energiebilanz in dieser Zeitschrift: I: Ludwig, 24,319 (1937); II: Kittel 28, 533 (1941); VIII: Hempel, 36,261 (1954).Erarbeitet an der obengenannten, von Prof. J. M. Pérès geleiteten Station an Schlechtwettertagen, die andere Untersuchungen unmöglich machten. Ihm und der Deutschen Forschungsgemeinschaft bin ich zu großem Danke verpflichtet.     

Autosomal dominant retinitis pigmentosa: Absence of the rhodopsin proline→histidine substitution (codon 23) in pedigrees from Europe

In exon 1 at codon 23 of the rhodopsin gene, a mutation resulting in a proline-to-histidine substitution has previously been observed in approximately 12% of American autosomal dominant retinitis pigmentosa (ADRP) patients. The region around the site of this mutation in the rhodopsin gene has been amplified and analyzed in affected individuals from 91 European ADRP pedigrees. The codon 23 mutation has been found to be absent in all cases, including a large Irish pedigree in which the disease gene has previously been shown to be closely linked to the rhodopsin locus. This indicates the presence of either allelic or nonallelic heterogeneity in ADRP.     

Inhibition of fungal growth by plant chitinases andβ-1,3-glucanases

Summary Plant chitinases and -1,3-glucanases have been demonstrated to inhibit fungal growth in model experiments, both on agar plates or in liquid media. Here,Trichoderma longibrachiatum was taken as a model to study the morphological changes caused by chitinase and glucanase treatments, using cytochemical techniques in combination with fluorescence and electron microscopy. Chitinase, alone or in the presence of glucanase, arrested growth of the hypha: it affected the extreme tip of the fungus producing a thinning of the wall, a balloon-like swelling and a rupture of the plasma membrane. Chitin and glucans were present in the wall, as shown by lectinand enzyme-binding experiments, but they had a different susceptibility to chitinase and -1,3-glucanase. Chitin was present at the apex and in the inner parts of the lateral walls; it was more susceptible to chitinase at the tip than in the subapical part. Glucans mostly occurred on the outer layer where they were degraded by glucanase. The latter did not affect the inner hyphal skeleton. It is suggested that the growth inhibition ofTrichoderma by hydrolytic enzymes is the consequence of a thinning of the cell wall in the hyphal apex, leading to an imbalance of turgor pressure and wall tension which causes the tip to swell and to burst.Abbreviations WGA-FITC wheat germ agglutinin labelled with fluorescein isothiocyanate - ConA-FITC concanavalin A labelled with fluorescein isothiocyanate - PEG polyethylene glycol - SEM scanning electron microscopy - TEM transmission electron microscopy     

Calibration of a resonance energy transfer imaging system.

A quantitative technique for the nondestructive visualization of nanometer scale intermolecular separations in a living system is described. A calibration procedure for the acquisition and analysis of resonance energy transfer (RET) image data is outlined. The factors limiting RET imaging of biological samples are discussed. Measurements required for the calibration include: (a) the spectral sensitivity of the image intensifier (or camera); (b) the transmission spectra of the emission filters; and (c) the quantum distribution functions of the energy transfer pair measured in situ. Resonance energy transfer imaging is demonstrated for two DNA specific dyes. The Förster critical distance for energy transfer between Hoechst 33342 (HO) and acridine orange (AO) is 4.5 +/- 0.7 nm. This distance is slightly greater than the distance of a single turn of the DNA helix (3.5 nm or approximately 10 base pairs), and is well below the optical diffraction limit. Timed sequences of intracellular energy transfer reveal nuclear structure, strikingly similar to that observed with confocal and electron microscopy, and may show the spatial distribution of eu- and hetero- chromatin in the interphase nuclei.     

Staphylococcus muscae, a new species isolated from flies.

A new coagulase-negative species of the genus Staphylococcus, Staphylococcus muscae, is described on the basis of the results of a study of four strains that were isolated from flies. 16S rRNA sequences of the type strains of S. muscae, Staphylococcus schleiferi, and Staphylococcus sciuri were determined and used, together with the corresponding sequences of Staphylococcus aureus and Staphylococcus epidermidis, for a comparative analysis. The new species is characterized taxonomically; this species is differentiated from the other novobiocin-susceptible staphylococci by its physiological and biochemical activities, cell wall composition, and levels of genetic relatedness. The type strain of this species is strain MB4 (= CCM 4175).     

Paracoccus denitrificans cytochrome c1 gene replacement mutants.

We describe the construction and characterization of gene replacement mutants for the respiratory chain component cytochrome c1 in the bacterium Paracoccus denitrificans. Its structural gene (fbcC) was inactivated by insertion of the kanamycin resistance gene, introduced into a suicide vector, and conjugated into Paracoccus; chromosomal mutants obtained by homologous recombination were selected by antibiotic resistance screening and further characterized biochemically. They showed the complete spectral, enzymatic, and immunological loss of the fbcC gene product together with a serious defect in the assembly of the two other gene products of the fbc operon, cytochrome b and the FeS protein. A possible role of the cytochrome c1 in the assembly process for the enzyme complex is discussed. A functional restoration to wild-type phenotype was achieved by complementing in trans with a newly constructed broad-host-range vector carrying the fbcC gene cassette. When the complete fbc operon was present on this vector, overexpression of complex III subunits was observed. Apart from their physiological significance, such mutants are a prerequisite for probing structure-function relationships by site-directed mutagenesis in order to understand molecular details of electron transport and energy transduction processes of this respiratory enzyme in bacteria and in mitochondria.     

Characterization of legiolysin (lly), responsible for haemolytic activity, colour production and fluorescence of Legionella pneumophila

A genomic library of Legionella pneumophila, the causative agent of Legionnaires' disease in humans was constructed in Escherichia coli K12 and the recombinant clones were tested for haemolysis and other phenotypic properties. Seven clones were identified which were able to confer haemolysis of human, sheep, and canine erythrocytes but which were unable to mediate proteolytic activities or cytotoxic effects on CHO- or Vero cells. Clones that exhibited this haemolytic property were also able to produce a brown colour and a yellow-green fluorescence activity detected on M9 plates containing tyrosine. The genetic determinant encoding these properties, termed legiolysin (lly) was mapped by Tn1000 mutagenesis and by subcloning experiments. Southern hybridization with an lly-specific gene probe showed that this determinant is part of the genome of L. pneumophila but is not identical to a protease gene of L. pneumophila which also mediates haemolysis. Minicell analysis of lly-specific plasmids exhibited a protein of 39 kDa. Polyclonal antibodies generated against a LacZ-Lly hybrid protein also recognized a 39 kDa protein produced either by the recombinant legiolysin-positive E. coli K12 clones or by L. pneumophila wild-type strains.     

Formation of lipid-bound oligosaccharides in yeast

Incubation of a membrane fraction from Saccharomyces cerevisiae with UDP-N-acetyl [¹⁴C] glucosamine catalyzes the tranfer of N-acetylglucosamine to an endeenous lipid fraction as well as a methanol-insoluble polymer. The glycolipid was shown to separate into three compounds by thin-layer chromatography. The biosynthesis of two of them could clearly be stimulated by the addition of dolichol monophosphate to the incubation mixture. Evidence is presented that the substances are dolichol pyrophosphate derivatives: dolichol pyrophosphate N-acetylglucosamine and dolichol pyrophosphate di-N-acetylchitobiose. The formation of the chitobiose-containing lipid was increased by reincubation of the glycolipid with non-radioactive UDP-N-acetylglucosamine.The same particulate preparation transferred mannose from GDPmannose to dolichol pyrophosphate di-N-acetylchitobiose, giving rise to a lipid-bound oligosaccharide. Molecular weight determination of the oligosaccharide moiety gave a value of 780, which is consistent with a tetrasaccharide containing two mannose subunits attached to di-N-acetylchitobiose.The methanol-insoluble radioactive product obtained in the presence of UDP-N-acetyl[¹⁴C]glucosamine was transformed by pronase treatment to a large extent into dialyzable material. It is suggested that the glycolipids described serve as intermediates in the glycosylation of yeast mannoproteins.     

Elektronenmikroskopische Untersuchungen anPaulinella chromatophora Lauterborn,einer Thekamöbe mit blau-grünen Endosymbionten (Cyanellen)

Zusammenfassung Die Feinstruktur der Cyanellen vonPaulinella chromatophora sowie die Bildung und Struktur der Kieselschuppen, die das Gehäuse dieser Thekamöbe aufbauen, wurden untersucht.Die beiden wurstförmigen Cyanellen besitzen eine 6–13 nm dicke Wandschicht. Sie liegen eingeschlossen in Vesikeln im Cytoplasma des Wirtes. Das Chromatoplasma der Cyanelle enthält 15–20 konzentrisch angeordnete Thylakoide, Plastoglobuli und Phycobilisomen. Das Centroplasma enthält polyedrische Körper.Das Gehäuse der Thekamöbe besteht aus verkieselten rechteckigen Schuppen, die sehr regelmäßig zum Gehäuse zusammengefügt sind. Die Schuppen haben eine komplizierte Feinstruktur. Sie entstehen, vielleicht unter Mitwirkung von Mikrotubuli, vor der Zellteilung in Vesikeln, die wahrscheinlich aus Zisternen des einzigen Dictyosomes der Thekamöbe hervorgehen. Dieses Dictyosom liegt dem Zellkern am aboralen Pol derPaulinella an. Hexagonale Körper (Virionen?) werden aus dem Zellkern und dem Cytoplasma des Wirtes beschrieben.

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Electron microscopical investigations onPaulinella chromatophora Lauterborn, a thecamoeba containing blue-green endosymbionts (cyanelles)

Summary The ultrastructure of the sausage-shaped cyanelles and the ultrastructure and formation of the thecal scales ofPaulinella chromatophora were investigated. The cyanelles have a 6–13 nm thick wall. They are lying within vesicles in the cytoplasma of the host. The chromatoplasma has 15–20 concentrically arranged thylakoids, plastoglobuli and phycobilisomes. The centroplasma contains polyhedral bodies. The theca ofPaulinella chromatophora is composed of rectangular scales arranged in a very regular manner. These scales exhibit a very complex ultrastructure. They are produced prior to cell division in large vesicles probably derived from cisternae of the only dictyosom which is located close to the nucleus in the aboral part of the thecamoeba. Microtubules may play a role in the morphogenesis of these scales.Hexagonal particles (virions?) are described from the nucleus and the cytoplasma of some of the thecamoebae.

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FräuleinBrigitte Schendel danke ich für ihre gute technische Mitarbeit, der Deutschen Forschungsgemeinschaft danke ich für Sachbeihilfe.     

Carbon monoxide and hydroxymercuribenzoate sensitivity of a fatty acid (omega-2) hydroxylase from Bacillus megaterium.

DNA-free minicells of $\text{Escherichia coli}$ will not allow growth of phage T-7, nor is RNA synthesis stimulated by phage infection. Thus, these miniature cells seem not to contain $\text{in vivo}$ RNA polymerase activity. However, DNA-dependent RNA polymerase activity can be unmasked in extracts with poly(dA-T) and Mn²⁺. This activity may represent a cytoplasmic pool of inactive RNA polymerase in normal cells.