Isolation of macrocyclic and non-macrocyclic trichothecenes (stachybotrys and fusarium toxins) from the Environment of 200 III Sport Horses

Satratoxins H and G, verrucarin J, and roridin E were isolated from the bedding straw of 200 sport horses exhibiting typical symptoms of stachybotryo-toxicosis. At the same time, the oat feed consumed by the horses contained non-macrocyclicFusarium trichothecenes: T-2 toxin and diacetoxyscirpenol.     

Heymann antibodies induce complement-dependent injury of rat glomerular visceral epithelial cells

The aim of this study was to investigate the in vitro role of the complement membrane attack complex (MAC) in the injury induced by nephritogenic anti-brush border vesicle (Fx1A) antibodies on rat glomerular visceral epithelial cells (GEC). Both sheep and rabbit anti-rat brush border vesicle IgG-induced complement-dependent lysis of cultured GEC. Fab fragments of sheep anti-rat brush border vesicles and polyclonal or monoclonal gp330 IgG were devoid of lytic activity. Shedding of cell-surface antigens induced by sheep or rabbit anti-rat brush border vesicle IgG protected GEC from subsequent exposure to lytic antibodies and complement, an effect that was not obtained with Fab fragments. When GEC were incubated with sheep or rabbit anti-rat brush border vesicle IgG in capping conditions, the C3 component was co-redistributed with Heymann immune complexes; in contrast, the MAC remained diffusely bound to the cell surface, indicating that it was not associated with the antigen-antibody complexes. The MAC was demonstrated on the surface of GEC by immunofluorescence staining with anti-MAC neoantigen and by electron microscopy of negatively stained membranes showing focal clusters of 110 A MAC lesions. When GEC were treated with sheep IgG or rabbit IgG plus C6-deficient sera, the cells were not lysed and MAC was not demonstrable on the surface; however, lytic activity was restored when C6-deficient sera were reconstituted with purified C6. The results are consistent with the interpretation that injury induced by Heymann antibodies on GEC is MAC-dependent.     

Enzymatic treatment of stickwater from fishmeal industry with the protease from Cucurbita ficifolia

Summary The protease fromC. ficifolia was applied to stickwater prior to evaporation. The evaporator was operated at reduced viscosities with an increase in solid concentration beyond 50% as compared to the untreated substrate.     

Addendum: Recovery of cellulases from spent broth of sugar beet pulp fermentation byTrichoderma reesei

Summary The exo- and endo-glucanases ofTrichoderma reesei were recovered after growth on sugar beet pulp by vacuum concentration followed by (NH₄)₂SO₄ or acetone precipitation. Final respective recoveries were 53% and 57% for the exoglucanase and 41% and 42% for the endoglucanase. The resuspended acetone precipitates of both enzymes showed no loss of activity after 30 d at 4°C, pH 4.8 and in the presence of 0.5% sorbic acid. (NH₄)₂SO₄-precipitated exoglucanase lost activity in the same period.

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Resumen Las exo y endoglucanasas deTrichoderma reesei se recuperaron después de su crecimiento en pulpa de remolacha azucarera mediante concentración al vacío seguida de precipitación con (NH₄)₂SO₄ o acetona. La recuperación final fue de 53% y 57% para la exoglucanasa y de 41% y 42% para la endoglucanasa. La resuspensión de los precipitados cetónicos de ambos enzimas no mostró perdidas de actividad después de 30 d a 4°C y pH 4.8 en presencia de ácido sórbico. La exoglucanasa precipitada con (NH₄)₂SO₄ perdió actividad durante este mismo periodo.

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Résumé Les exo- et endoglucanases deTrichoderma reesei ont été récupérées après croissance sur pulpe de betterave sucrière, par concentration sous vide après précipitation par le sulfate ammonique ou l'acétone. Les récupérations ont été respectivement de 53% et 57% pour l'exoglucanase et de 41% et 42% pour l'endoglucanase. Les resuspensions des deux enzymes après précipitation à l'acétone n'ont révélé aucune perte d'activité après 30 jours à 4°C, pH 4.8 et en présence de 0.5% d'acide sorbique. L'exoglucanase précipitée au sulfate ammonique avait perdu toute activité dans le même laps de temps.

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(Based on a paper presented at the First Latin American Congress on Biotechnology, Tucumán, Argentina, October 4–8, 1987)     

Cytology of induced systemic resistance of cucumber to Colletotrichum lagenarium

The infection of cucumber leaves by Colletotrichum lagenarium was studied using cytological methods. Its progress in untreated plants was compared with that in plants in which systemic resistance had been induced by pre-infecting the first true leaf with the same fungus. In induced plants, a reduction of fungal development was observed at the leaf surface, in the epidermis, and in the mesophyll. On the leaf surface, formation of appressoria was slightly reduced. In the epidermis, enhanced formation of papillae beneath appressoria, and possibly increased lignification of entire cells, correlated with reduced development of infection hyphae. Papillae contained callose, identified by staining with aniline-blue fluorochrome and digestion with -1,3-glucanase, as a main structural component. In the mesophyll, reduced fungal development provided evidence for the existence of an additional induced defence reaction. The results imply that preinfection elicited a systemic, multicomponent defence reaction of the host plant against the fungus.Dedicated to the memory of Professor H. Grisebach     

Covalent immobilization of avidin on glassy carbon electrodes as the basis for multivalent biosensors.

One of the most crucial steps for the successful construction of a biosensor is the appropriate and reproducible coupling of the biological part (e.g. enzyme, antibody) to the inorganic moiety of the device (e.g. electrode, microchip). In this paper three methods of immobilization of avidin to a glassy carbon electrode are described. Depending on the type of immobilization, avidin may lose its biological activity as determined by an enzyme immunoassay, using biotinylated reagents. If avidin is covalently bound to the glassy carbon electrode via the bridge molecule 4.4'-diaminodiphenylamine, the biological activity is retained. About 1.5 pmol of avidin can be bound to the electrode (3 mm in diameter), resulting in a nearly complete monolayer of protein.     

Evidence for the presence of muscarinic acetylcholine receptors in bovine brain coated vesicles

Evidence obtained from quinuclidinylbenzilate binding determinations suggested that muscarinic acetylcholine receptor molecules are present in purified bovine brain coated vesicles. Immunoprecipitates formed from coated vesicles with polyclonal antibodies to clathrin bound on the surface of fixed Staphylococcus aureus cells also showed quinuclidinylbenzilate binding activity. The high purity of coated vesicles was established by assays for biochemical markers and by electron microscopy. Muscarinic receptor sites for quinuclidinylbenzilate binding to coated vesicles displayed a Kd of 25 pM and a Bmax of about 191 fmol/mg of protein. Binding competition experiments using atropine, N-methylscopolamine, oxotremorine, and carbamylcholine confirmed the typical muscarinic nature of the binding site. Ranking order of potency for the receptors was: atropine greater than N-methylscopolamine greater than oxotremorine greater than carbachol Analysis of data using a two-site model revealed 13% high-affinity sites for oxotremorine, 66% high-affinity sites for carbachol, and 62% for the antagonist N-methylscopolamine. Heterogeneity of binding affinities for muscarinic drugs detected in the coated vesicles may be related to the presence of coated vesicle subpopulations in brain tissue, (Kohtz, D. S., Kohtz, J. D., Schook, W. J., and Puszkin, S. (1985) J. Cell Biol. 101, 48a; Pfeffer, S. R., and Kelly, R. B. (1985) Cell 40, 949-957).     

Mikropinozytose im Zentralnervensystem

Zusammenfassung An durch Perfusion mit Glutaraldehyd fixierten Rattengehirnen wurde das Erscheinungsbild der Mikropinozytose in Elementen der Meso- und Neuroglia sowie an den Perikarya und synaptischen Endformationen der Nervenzellen elektronenmikroskopisch dargestellt.Die bei der Mikropinozytose von der Zellmembran invaginierten Caveolen und Tubuli können einfache Verzweigungen zeigen. Ihre Oberfläche und die der mikropinozytotischen Bläschen zeigen an der gegen das Zytoplasma gerichteten Membranseite einen Stachelsaum. Diese Membrandifferenzierung dürfte mit der Resorption besonderer, zum Teil makromolekularer Substanzen zusammenhängen.Im Bereich großer Synapsen, z.B. in den Moosfasertelodendren der Glomerula cerebellaria oder in der Zona glomerulosa des Bulbus olfactorius sind mikropinozytotische Invaginationen und Bläschen sehr häufig. Möglicherweise übernehmen sie von den postsynaptischen Dendriten, die dünne Zytoplasmaprotrusionen in die Invaginationen hineinsenden, Stoffe. Es wird vermutet, daß es sich hierbei um inaktivierte Transmittersubstanz handelt, die auf diesem Wege dem präsynaptischen Abschnitt wieder zugeführt wird. Die zurückresorbierten Abbauprodukte der Transmittersubstanz werden in einem präsynaptischen Golgi-Komplex resynthetisiert und in synaptischen Bläschen angereichert. Dieses morphologische Bild ergänzt die biochemische Hypothese eines Acetylcholin-Kreislaufes im Bereich von Nervenendigungen.Entsprechende mikropinozytotische Erscheinungen wurden in caudalen Abdominalganglien von Leucophaea maderae beobachtet. Es wird angenommen, daß die Mikropinozytose ein allgemein verbreiteter Resorptionsmechanismus im Zentralnervensystem ist.Herrn Prof. Dr. F. Wassermann zum 80. Geburtstag gewidmet.Mit dankenswerter Unterstützung durch die Deutsche Forschungsgemeinschaft.