Zum mikrochemischen Nachweis der Betaine in der Pflanze

Ohne Zusammenfassung     

The Bacillus subtilis Nucleotidyltransferase Is a tRNA CCA-Adding Enzyme

There has been increased interest in bacterial polyadenylation with the recent demonstration that 3′ poly(A) tails are involved in RNA degradation. Poly(A) polymerase I (PAP I) of Escherichia coli is a member of the nucleotidyltransferase (Ntr) family that includes the functionally related tRNA CCA-adding enzymes. Thirty members of the Ntr family were detected in a search of the current database of eubacterial genomic sequences. Gram-negative organisms from the β and γ subdivisions of the purple bacteria have two genes encoding putative Ntr proteins, and it was possible to predict their activities as either PAP or CCA adding by sequence comparisons with the E. coli homologues. Prediction of the functions of proteins encoded by the genes from more distantly related bacteria was not reliable. The Bacillus subtilis papS gene encodes a protein that was predicted to have PAP activity. We have overexpressed and characterized this protein, demonstrating that it is a tRNA nucleotidyltransferase. We suggest that the papS gene should be renamed cca, following the notation for its E. coli counterpart. The available evidence indicates that cca is the only gene encoding an Ntr protein, despite previous suggestions that B. subtilis has a PAP similar to E. coli PAP I. Thus, the activity involved in RNA 3′ polyadenylation in the gram-positive bacteria apparently resides in an enzyme distinct from its counterpart in gram-negative bacteria.     

The functional and structural borders between the CSF-and blood-dominated milieus in the choroid plexuses and the area postrema of the rat

Summary In the borderline area between the hemal milieu of the choroid plexuses (PC) and the interstitial cerebrospinal-fluid (CSF) compartment, ground substances displaying increased amounts of basal lamina-like material and containing negatively charged sulfated glycosaminoglycans appear to be endowed with selective properties. They may function as a sieve or filtration barrier gradually controlling the passage of substances between the two milieus, depending on their charge and molecular weight. Special structural features and functional properties of ependymal cells are associated with such bordering structures. These ependymal cells are transitional elements between choroid epithelium and ciliated ependymal cells. As judged from experiments with horseradish peroxidase and conventional electron microscopy, occluding junctions at the basal pole of these cells prevent a rapid alteration in the milieu conditions, enabling gradual change from hemal to CSF composition near the bases of these transitional ependymal cells. The borderline structures between the hemal milieu of the PC and the area postrema (1) are established by leptomeningeal cells which face a hemal mileu, (2) are endowed with conspicuous tight junctions, and (3) produce a flocculent substance, the light-microscopic equivalent of which is PAS positive. These structures probably establish an effective barrier between the two milieus of different composition. The functional characteristics and the morphology of the meningeal cells facing the hemal milieu of neurohemal regions resemble closely the neurothelial cells, which are interposed between the CSF milieu and the hemal milieu in the dura mater. The present results suggest that the location between the hemal and the CSF milieu is decisive for the transformation of leptomeningeal cells into neurothelial elements.Supported by the Deutsche Forschungsgemeinschaft (Grant Kr 569/5-1)Dedicated to Professor Dr. med., Dr. med. vet. h.c., Dr. phil. h.c. Andreas Oksche on the occasion of his 60th birthday     

Phage-Antibiotic Synergy (PAS): beta-lactam and quinolone antibiotics stimulate virulent phage growth

Although the multiplication of bacteriophages (phages) has a substantial impact on the biosphere, comparatively little is known about how the external environment affects phage production. Here we report that sub-lethal concentrations of certain antibiotics can substantially stimulate the host bacterial cell's production of some virulent phage. For example, a low dosage of cefotaxime, a cephalosporin, increased an uropathogenic Escherichia coli strain's production of the phage PhiMFP by more than 7-fold. We name this phenomenon Phage-Antibiotic Synergy (PAS). A related effect was observed in diverse host-phage systems, including the T4-like phages, with beta-lactam and quinolone antibiotics, as well as mitomycin C. A common characteristic of these antibiotics is that they inhibit bacterial cell division and trigger the SOS system. We therefore examined the PAS effect within the context of the bacterial SOS and filamentation responses. We found that the PAS effect appears SOS-independent and is primarily a consequence of cellular filamentation; it is mimicked by cells that constitutively filament. The fact that completely unrelated phages manifest this phenomenon suggests that it confers an important and general advantage to the phages.     

The presence of N-acetylneuraminic acid in Malpighian tubules of larvae of the cicada Philaenus spumarius

Sialic acid-containing glycoconjugates are generally considered to be unique to the deuterostomes, a lineage of the animal kingdom which includes animals from the echinoderms up to the vertebrates. There are, however, two isolated reports of sialic acid occurring in the insect species Drosophila melanogaster and Galleria mellonella. Since insects are classified as protostomes, these findings call previous assumption on the phylogenetic distribution and thus on the evolution of sialic acids into question. Here, we report the occurrence of N-acetylneuraminic acid (Neu5Ac) in larvae of the cicada Philaenus spumarius. Cytochemical analysis of larval sections with lectins from Sambucus nigra and Limax flavus suggested the presence of sialic acids in the concrement vacuoles of the Malpighian tubules. The monoclonal antibody MAb 735, which is specific for polysialic acid, labelled the same structures. A chemical analysis performed by HPLC of fluorescent derivatives of sialic acids and by GLC-MS provided sound evidence for the presence of Neu5Ac in the Philaenus spumarius larvae. These data suggest that in this cicada Neu5Ac occurs in 2,8-linked polysialic acid structures and in 2,6-linkages. The results provide further evidence for the existence of sialic acids in insects and in linkages known to occur in glycoconjugates of deuterostomate origin.     

Characterization of an extracellular laccase of Leptosphaerulina chartarum

Laccase-producing fungi were isolated from air, using selective media with a chromogenic substrate to indicate enzyme activity. The best laccase producer strain proved to be a Leptosphaerulina chartarum isolate. Laccase production was investigated in the presence of various inducers in different cultivation conditions. The extracellular laccase was purified for further investigations. SDS-PAGE showed that this laccase is a monomeric protein of 38 kDa molecular weight. The enzyme is active in the pH-range of 3.5–6, with an optimum at pH 3.8. It is active in the 10–60 °C temperature range, with an optimum at 40 °C. After 20 min incubation at temperatures above 70 °C the enzyme lost its activity. Degradation of seven aniline and phenol compounds (2,4-dichlorophenol; 2-methyl-4-chlorophenol; 3-chloroaniline; 4-chloroaniline; 2,6-dimethylaniline; 3,4-dichloroaniline and 3-chloro-4-methylaniline) was investigated, with or without guaiacol (2-methoxyphenol) as mediator molecule. Addition of a mediator to the system significantly increased the degradation levels. These results confirmed that the isolated laccase is able to convert these harmful xenobiotics at in vitro conditions.     

Somatostatin-immunoreactive fiber projections into the brain stem and the spinal cord of the rat

Summary By use of the PAP-immunohistochemical staining technique with serial sections, somatostatin-immunoreactive fiber projections into the brain stem and the spinal cord are described. These projections originate in the periventricular somatostatin-immunoreactive perikarya of the hypothalamus and form three main pathways: (1) along the stria medullaris thalami and the fasciculus retroflexus into the interpeduncular nucleus; (2) along the medial forebrain bundle into the mammillary body; and (3) via the periventricular gray and the bundle of Schütz into the midbrain tegmentum. Densely arranged immunoreactive fibers and/or basket-like fiber terminals are observed within the following afferent systems: somatic afferent systems (nucleus spinalis nervi trigemini, substantia gelatinosa dorsalis of the entire spinal cord), and visceral afferent systems (nucleus solitarius, regio intermediolateralis and substantia gelatinosa of the sacral spinal cord). These projections form terminals around the perikarya of the second afferent neuron. Perikarya of the third afferent neuron are influenced by somatostatin-immunoreactive projections into the auditory system (nucleus dorsalis lemnisci lateralis, nucleus corporis trapezoidei). Furthermore, a somatostatin-immunoreactive fiber projection is found in the ventral part of the medial accessory olivary nucleus, in nuclei of the limbic system (nucleus habenularis medialis, nuclei supramamillaris and mamillaris lateralis) and in the formatio reticularis (nucleus Darkschewitsch, nuclei tegmenti lateralis and centralis, nucleus parabrachialis lateralis, as well as individual perikarya of the reticular formation). Targets of these projections are interneurons within interlocking neuronal chains.Supported by the Deutsche Forschungsgemeinschaft (Grant Nr. Kr 569/3) and Stiftung Volkswagenwerk     

Isolation and Characterization of Antagonistic Bacillus Strains Capable to Degrade Ethylenethiourea

In this study, more than 150 bacteria showing antagonistic properties against bacterial and fungal pathogens of the tomato plant were isolated and characterized. The most efficient agents against these phytopathogenic microorganisms belong to the genus Bacillus: the best biocontrol isolates were representatives of Bacillus subtilis, B. mojavensis and B. amyloliquefaciens species. They intensively produced fengycin or/and surfactin depsipeptide antibiotics and also proved to be excellent protease secretors. It was proved, that the selected strains were able to use ethylenethiourea (ETU) as sole nitrogen source. These antagonistic and ETU-degrading Bacillus strains can be applied as biocontrol and also as bioremediation agents.     

Anticandidal effect of berry juices and extracts from Ribes species

The biological activities of fruit juices and pomace (skin, seeds) extracts from blackcurrant (Ribes nigrum), gooseberry (Ribes uva-crispa) and their hybrid plant (jostaberry, Ribes × nidigrolaria) were evaluated against the most frequently isolated twelve human pathogenic Candida species by broth dilution tests. The phenolic content of juice, water and methanol extracts were measured and the relationship with antifungal activity was assessed. Growth of the most Candida species was inhibited, with the exception of C. albicans, C. krusei, C. lusitaniae and C. pulcherrima. R. nigrum, with the highest phenol content, was observed to have the highest anticandidal activity, indicating a positive correlation between phenol content and antifungal activity.