Studies on the Movement of Water through the Isolated Toad Bladder and Its Modification by Vasopressin

Measurements of diffusion permeability and of net transfer of water have been made across the isolated urinary bladder of the toad, Bufo marinus, and the effects thereon of mammalian neurohypophyseal hormone have been examined. In the absence of a transmembrane osmotic gradient, vasopressin increases the unidirectional flux of water from a mean of 340 to a mean of 570 µl per cm² per hour but the net water movement remains essentially zero. In the presence of an osmotic gradient but without hormone net transfer of water remains very small. On addition of hormone large net fluxes of water occur; the magnitude of which is linearly proportional to the osmotic gradient. The action of the hormone on movement of water is not dependent on the presence of sodium or on active transport of sodium. Comparison of the net transport of water and of unidirectional diffusion permeability of the membrane to water indicates that non-diffusional transport must predominate as the means by which net movement occurs in the presence of an osmotic gradient. An action of the hormone on the mucosal surface of the bladder wall is demonstrated. The effects of the hormone on water movement are most simply explained as an action to increase the permeability and porosity of the mucosal surface of the membrane.     

Feinstruktur und Histochemie der Sexualduftdrüse des Seidenspinners Bombyx mori L.

Zusammenfassung Das Weibchen des Seidenspinners, Bombyx mori L., erzeugt zur Anlockung der männlichen Artgenossen in paarigen, ausstülpbaren Drüsen, den am Abdomenende gelegenen Sacculi laterales, einen spezifischen Sexuallockstoff. Dieser Lockstoff, das Bombykol, ist in seiner chemischen Konstitution bekannt und auch in synthetischer Form verfügbar.Das Drüsenepithel stellt eine differenzierte Form der normalen Insekten-epidermis dar. Wie diese besteht es aus einer einschichtigen Zellage, die an ihrer Außenfläche eine chitinhaltige Cuticula und innen, an der Grenze zum Hämolymphraum, eine Basalmembran trägt. Laterale Verzahnungen (Interdigitationen) und Desmosomen sichern den Zusammenhalt der Zellen, die beim Aus- und Einstülpen der Drüse starken Formveränderungen ausgesetzt sind.Die Zellen enthalten große, gelappte Zellkerne mit sehr locker strukturiertem Chromatin; im Cytoplasma ist ein agranuläres endoplasmatisches Reticulum stark ausgeprägt, das mit dem Ansteigen der Lockaktivität an die Stelle eines granulären endoplasmatischen Reticulums tritt. Der Golgi-Apparat ist nur unscheinbar; Mitochondrien sind in großer Zahl vorhanden.Im Gegensatz zur undifferenzierten Epidermis treten im Drüsenepithel mit Beginn der Lockaktivität in zunehmendem Maße Lipidtröpfchen auf. In diesen wird auf Grund histologischer und histochemischer Befunde eine Vorstufe des Lockstoffes vermutet.Die Grenzfläche der Zelle zur Cuticula ist durch Ausbildung eines Falten-saums 30–60fach vergrößert. Dieser wird von lamellenartigen Zellvorsprüngen gebildet, die sehr dicht stehen und weitgehend parallel zueinander verlaufen.Die Ausbildung des Faltensaums kann mit dem Anstieg der Lockwirkung der Drüse korreliert werden. Es wird ein Zusammenhang zwischen der Vergrößerung der apikalen Zelloberfläche und der Lockstoffsekretion vermutet.Das Drüsenepithel unterscheidet sich von der Intersegmentalmembran durch eine bedeutend stärkere Aktivität der NADP-Tetrazolium-Reduktase (früher als TPN-Diaphorase bezeichnet), was mit der stärkeren Synthesetätigkeit der Drüsenzellen in Zusammenhang gebracht wird.Der Weg des Lockstoffs durch die Zellmembran und die Cuticula konnte nicht verfolgt werden. Die Cytoplasmamembran bleibt stets intakt; die Cuticula läßt keine Kanalbildungen erkennen. Es wird vermutet, daß sich die Absonderung des Lockstoffs auf molekularer Ebene abspielt.Herrn Priv.-Doz. Dr. D. Schneider danke ich für die Anregung und stete Förderung der Arbeit, Herrn Prof. Dr. G. Peters für die Überlassung eines Arbeitsplatzes, den Herren Priv.-Doz. Dr. Dr. H. Hager und Dr. K. Blinzinger (Abteilung für Neurozytologie) und Dr. G. Kreutzberg (Hirnpathologisches Institut) für fördernde Kritik und technische Unterstützung.Dissertation der Naturwissenschaftlichen Fakultät der Universität München.     

Assay of substances stimulatory to legume nodule formation

Two methods were developed for the assay of substances stimulatory to the nodulation of bean (Phaseolus vulgaris) roots growing from segments of hypocotyl tissue. Coconut water was the chief source of active material, but extracts of cotyledons, hypocotyls and leaves of beans and of horse chestnut fruits were also stimulatory. High concentrations of nitrate improved nodulation both in the presence and absence of coconut water. The ash of coconut water was inactive. Whole alfalfa seedlings formed nodules in the dark when grown in the split medium, but nodulation was not improved by the addition of coconut water.     

Partial purification of a legume nodulation factor present in coconut water

The nodulation of adventitious roots growing from segments of bean hypocotyl tissue was used as a bioassay for the material present in coconut water which stimulated nodulation. The active material in coconut water is acidic, but it was not possible to extract it from an acid solution with organic solvents. A purification of approximately 70-fold (on a dry wt basis) was obtained using activated charcoal, but at least 10 different compounds were present in the active fractions. A purified fraction of coconut water, which is stimulatory to the growth of carrot root explants, was active in the nodulation assay at a concentration of 2 μg/ml. This represents a 4000-fold purification of the diffusible fraction of coconut water. The charcoal fractionation procedure can be applied to the active material present in extracts of bean leaves.     

Permeability of Nitrobacter agilis to Organic Compounds.

Ida, S. (Cornell University, Ithaca, N.Y.), and M. Alexander. Permeability of Nitrobacter agilis to organic compounds. J. Bacteriol. 90:151-156. 1965.-None of a variety of inorganic ions or organic compounds served as a sole energy source for the growth of Nitrobacter agilis, and the test substrates were not oxidized by either intact cells or extracts of the obligate chemoautotroph. The organic substances did not serve as sole carbon sources for the bacterium in a synthetic medium, and they failed to enhance the rate of nitrite oxidation. The organism was permeable to acetate and a number of other simple carbon compounds, however, and exogenously supplied acetate was converted to a number of products. On the basis of these findings, possible reasons are examined for the inability of the chemoautotroph to use exogenous organic compounds as energy or carbon sources.     

Competition between soil bacteria and fusarium

Summary The phenomenon of competition has been characterized in liquid medium and sterile soil systems using a variety of soil bacteria andFusarium oxysporum f.cubense as test organisms. For most of the bacteria, suppression of the fungus was the result of a biologically induced nitrogen deficiency, this effect being reversed by the addition of excess inorganic nitrogen. High populations of competitors were found in two soils of neutral pH, but no isolates competed in the acid San Alejo loam.Agrobacterium radiobacter was able to compete when San Alejo loam was limed to about pH 6.6. Inhibition of the fungus by a number of gram-positive, spore-forming rods could not be accounted for in terms of competition for nutrients or by antibiotic production in artificial media.The competitive ability ofA. radiobacter when tested in twelve Central American soils was found to be related to pH in acid and neutral environments but was correlated with texture, organic-matter content and total nitrogen in soils of intermediate pH. In all soils where inhibition occurred, the competitive effect was overcome by additions of inorganic nitrogen. Excluding the group ofBacillus spp., the competitive ability of soil bacteria was related to the ability to develop in the absence of amino acids and growth factors but could not be correlated with growth rates of the bacteria in soil or liquid medium.It is suggested that competition for nutrients is a significant means of ecological control among members of the soil microflora and, together with competitive interactions for space and oxygen, may be the major factors governing the biological control of soil-borne fungi.The investigation was supported in part by a grant from the United Fruit Company. Agronomy Paper No. 471.