Induction of alternative oxidase synthesis by herbicides inhibiting branched-chain amino acid synthesis

Sycamore suspension cells ( Acer pseudoplatanus L.) were incubated in the presence of sulfonylurea and imidazolinone herbicides. These inhibitors of acetolactate synthase (ALS), a key enzyme of branched-chain amino acid synthesis, triggered a dramatic induction of the alternative oxidase (AOX). AOX activity increased in treated cells, eventually exceeding cytochrome (cyt) pathway activity. This induction of AOX activity was correlated with the accumulation of a 35 kDa AOX protein in isolated mitochondria, detected by Western blotting with a monoclonal antibody against Sauromatum guttatum AOX. It was preceded by the accumulation of putative 1.6 kb AOX mRNA, detected using an Aox cDNA probe from soybean. The metabolic perturbations induced by the herbicides rather than the herbicide molecules themselves were responsible for this induction of AOX. However, α-oxobutyrate (one of the substrates of ALS) and its transamination product, α-aminobutyrate, which accumulated after herbicide treatment, were not involved. The inhibition of branched-chain amino acid synthesis was probably somehow responsible for the AOX induction since: (i) a mixture of those amino acids (leucine, isoleucine, valine) prevented AOX induction by ALS inhibitors; (ii) the herbicide Hoe 704, a potent inhibitor of acetolactate reducto-isomerase (the enzyme following ALS in the branched-chain amino acid pathway), also triggered AOX induction.     

Auswirkungen von Hochspannungstrassen auf die Fl?chennutzung überwinternder Bl??- und Saatg?nseAnser albifrons, A. fabalis

Zusammenfassung  Während das Mortalitätsrisiko für Vögel durch Drahtanflug und Stromschlag an Hochspannungsleitungen vielfach untersucht wurde, könnte auch ein Einfluß von Hochspannungstrassen (110 kV) auf Vögel bestehen, die sich am Boden aufhalten. Dieser wurde 1994/95 am Beispiel der Flächennutzung überwinternder Bläß- und Saatgänse (Anser albifrons, A. fabalis) am Unteren Niederrhein untersucht. Da Gänse offene ungestörte Äsungsflächen beanspruchen, wurde der Arbeitshypothese nachgegangen, daß trassennahe Bereiche von Gänsen gemieden bzw. vermindert genutzt werden. Die Besuchsfrequenz von vier häufig genutzten trassenüberspannten Äsungsflächen wurde ermittelt. Im Anschluß an die einzelnen mehrtägigen Beäsungsphasen wurde die Losungsdichte (LD) als Maß der Flächennutzung senkrecht zum Trassenverlauf bestimmt. An 16–20 m hohen Trassen stieg die LD mit zunehmender Entfernung zur Trasse. Eine signifikante Abnahme der LD trat unterhalb von 40–80 m Abstand zur Trasse ein. Kleine Teilflächen, die durch die Trassen von größeren Bereichen abgetrennt sind, wurden weniger genutzt. An einer 60 m hohen, den Rhein Überquerenden Trasse, konnte kein trassenbedingter Unterschied der LD festgestellt werden. Mögliche Ursachen für die beschriebenen Effekte werden diskutiert.

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The impact of power lines on field selection and grazing intensity of wintering White-fronted- and Bean GeeseAnser albifrons, A. fabalis

In addition to studies about the mortality risk by collision and electrocution power lines may have an impact on birds during foraging behaviour on the ground. We examined this question in geese (Anser albfrons, A. fabalis) wintering in the lower Rhine area of North Rhine Westfalia (FRG) 1994/95. Because geese prefer large areas with low levels of disturbance by human activities or urbanic structures like buildings or roads, we hypothesized that they might be disturbed by the presence of power lines in their foraging sites and thus avoid areas next to the lines. To quantify the grazing intensity in relation to the distance from the power lines (110 kV), dropping densities have been repeatedly counted after independant grazing intervals in four different feeding sites. Our data show that: 1. Grazing intensity increases with the distance to power lines of low height. Significantly reduced grazing levels have been found in distances less than 80 to 40 m to the power lines. 2. No effects have been observed for a power line in an area next to the river rhine where the wires crossed the ground in a height of about 60 metres. 3. In small pasture or field areas, cut off by power lines from large ones, a generally reduced grazing amount is evident. Several conceivable reasons for the described effects are discussed.

     

DNA sequence analysis of mitochondrial Cyt-b and the species status ofLaniarius dubiosus (Rchw. 1899)

In 1899Reichenow described an african bushshrike in juvenile plumage as a new species. He named itLaniarius dubiosus. DNA from type material (feathers and skin) was extracted and DNA sequences from the mitochondrial Cyt-b gene were analysed. Comparisons of DNA-sequences from other bushshrikes (including the type-specimen from Luehder's bushshrikeLaniarius lühderi) support the judgement that dubiosus does not represent a full species rather than a representative of the western subspecies of the Luehder's bushshrikeLaniarius luehderi.

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Zusammenfassung Der vonReichenow (1899) als neue Art beschriebene, sich im Jugendkleid befindliche BuschwürgerLaniarius dubiosus ist nach DNA-Sequenzanalysen des mitochondrialen Cyt-b Genes mit hoher Wahrscheinlichkeit ein Jungvogel der westlichen Rasse des BraunscheitelwürgersLaniarius lühderi. Auf der Basis eines DNA-Stammbaumes von sieben verschiedenen Buschwürgern ist der 1991 neu beschriebeneLaniarius liberatus am wenigsten eng mitL. dubiosus verwandt.

     

Recent approaches to probe functional groups in ribonuclease P RNA by modification interference

Modification interference is a powerful method to identify important functional groups in RNA molecules. We review here recent developments of techniques to screen for chemical modifications that interfere with (i) binding of(pre-)tRNA to bacterial RNase P RNA or (ii) pre-tRNA cleavage by this ribozyme. For example, two studies have analyzed positions at which a substitution of sulfur for thepro-Rp oxygen affects tRNA binding [1] or catalysis [2]. The results emphasize the functional key role of a central core element present in all known RNase P RNA subunits. The four sulfur substitutions identified in one study [2] to inhibit the catalytic step also interfered with binding of tRNA toE. coli RNase P RNA [1]. This suggests that losses in binding energy due to the modification at these positions affect the enzyme-substrate and the enzyme-transition state complex. In addition, the two studies have revealed, for the first time, sites of direct metal ion coordination in RNase P RNA. The potentials, limitations and interpretational ambiguities of modification interference experiments as well as factors influencing their outcome are discussed.Abbreviations nt nucleotide(s) - PAGE polyacrylamide gel electrophoresis     

Comparison of the main siderophores produced by some species of Streptomyces

The production of siderophores by four Streptomyces strains, S. ambofaciens, S. coelicolor, S. lividans, and S. viridosporus, was studied under iron-limited conditions. S. viridosporus produced two different siderophores: the linear desferrioxamine B and the cyclic desferrioxamine E. The latter was produced by the other strains and was the main siderophore of S. ambofaciens. The linear desferrioxamine G was the major form of S. coelicolor and S. lividans. The uptake rates of ⁵⁵Fe-labeled ferrioxamines by S. lividans and S. viridosporus showed that the G form was incorporated less efficiently than the B and E forms.     

Isolation of a YAC clone covering a cluster of nine S100 genes on human chromosome 1q21: rationale for a new nomenclature of the S100 calcium-binding protein family

S100 proteins are low-molecular-weight calcium-binding proteins of the EF-hand superfamily and appear to be involved in the regulation of a number of cellular processes such as cell cycle progression and differentiation. More than 10 members of the S100 protein family have been described from human sources so far. We have now isolated a YAC clone from human chromosome 1q21, on which 9 different genes coding for S100 calcium-binding proteins could be localized. Moreover, we have mapped the gene coding for S100P to human chromosome 4p16 and thereby completed the chromosomal assignments of all known human S100 genes. The clustered organization of S100 genes in the 1q21 region allows us to introduce a new logical nomenclature for these genes, which is based on the physical arrangement on the chromosome. The new nomenclature should facilitate and further the understanding of this protein family and be easily expandable to other species.     

Glucose as an auxiliary substrate

Summary A theoretical consideration is presented of the comparative efficiency of carbon conversion of glucose by the Embden-Meyerhof-Parnas (EMP) and the oxidative hexosemonophosphate (HMP) pathways. As a result it is shown that maximum carbon conversion, that is 89%, is possible when glucose is assimilated via the EMP pathway. This value is diminished in proportion to the participation of the HMP pathway in carbon assimilation and is halved when glucose is incorporated entirely via this pathway. If NADPH is included as a source of energy, glucose may behave both as an excess carbon and an excess energy substrate, the latter being the case when greater portions of the HMP pathway operate, and the extent of this is in turn dependent on the P/O quotient. If NADPH cannot be used for ATP synthesis, glucose remains an excess carbon substrate throughout, although when the HMP pathway accounts for more than 26% of glucose assimilation an increasing excess of reduction equivalents is produced. These results are interpreted in terms of mixed-substrate utilization for improving growth yield when glucose is to be used as the excess carbon component.     

Identification of Merkel cells in human skin by specific cytokeratin antibodies:

Merkel cells are special neurosecretory cells which, in adult human skin, are usually very scarce. By immunofluorescence microscopy using antibodies to human cytokeratin polypeptide no. 18, we localized distinct non-keratinocyte cells in the glandular ridges of human fetal and adult plantar epidermis. Using electron and immunofluorescence microscopy, these cells were identified as Merkel cells containing typical neurosecretory granules as well as bundles of intermediate-sized filaments and desmosomes. Two-dimensional gel electrophoresis of the cytoskeletal fractions of microdissected epidermal preparations highly enriched in Merkel cells indicated the presence of cytokeratin polypeptides nos. 8, 18 and 19 which are typical of diverse simple epithelia of the human body. Double immunofluorescence microscopy showed that these human Merkel cells contain neither neurofilaments nor vimentin filaments. In human fetuses of 18-24 weeks of age, conspicuously high concentrations of Merkel cells, reaching a density of approximately 1,700 Merkel cells/mm2 skin, were found in the glandular ridges of plantar skin. The concentration decreased considerably at newborn and adult stages. Thin cell processes (up to 20 microns long) were observed in many fetal epidermal Merkel cells. In addition, we detected isolated Merkel cells deeper in the dermis (i.e. at distances of, at most, 100 microns from the epidermis) in fetal and newborn plantar skin. Our results show that Merkel cells are true epithelial cells which, however, differ profoundly from epidermal keratinocytes in their cytokeratin expression. The findings are discussed in relation to the much disputed question of the origin of Merkel cells. The present data speak against the immigration of Merkel cells from the neural crest, but rather suggest that they originate from epithelial cells of the skin, although most probably not from differentiated keratinocytes.     

Peroxidases in Acetabularia: their possible role in development

Abstract. Crude enzymatic extracts from Acetabularia exhibit very low peroxidase activity after a lag period. Starch gel electrophoresis of extracts from growing algae shows a single, extremely anodic band. Extracts of small, slow-growing or cap-bearing algae, which do not grow any more, do not exhibit any peroxidase band. Cytochemical staining with benzidine reveals changes in both the quantity and distribution of peroxidase along the polarized Acetabularia cell. The homogenous staining of small algae becomes distributed along a negative apico-basal gradient when the algae initiate their rapid growth phase. This polarized pattern is repeated on the hair whorls. A similar developmental sequence directs cap growth, with an initial intense staining reaction of the primordium, which later leaves only the corona inferior stained blue. Finally, the Acetabularia cell remains slightly blue at the edges of the rhizoidal out-growths and cap rays. Crude extracts of Acetabularia induce a lag in standard horseradish peroxidase (HRP) activity. The inhibitor is always present in small and growing algae; it is sometimes absent or less active in cap-bearing algae. In no case does it change the kinetics of the HRP reaction with guaïacol. The lag is completely suppressed by pretreatment with either H₂O₂ or ascorbate oxidase. The changes in peroxidase activity, correlated with developmental stage and according to a polarized gradient, suggest that the enzyme could be involved in some way in the control of morphogenesis in Acetabularia . An inhibitor of peroxidase activity, which disappears as the cap matures, might, in turn, exert a regulatory function.