Degradation of Coal by the Fungi Polyporus versicolor and Poria monticola

We report that two species of basidiomycete fungi (Polyporus versicolor and Poria monticola) grow in minimal liquid or solid medium when supplemented with crushed lignite coal. The fungi also grow directly on crushed lignite coal. The growth of both fungi was observed qualitatively as the production and extension of hyphae. No fungal growth occurred in minimal agar medium without coal. The fungi degraded solid lignite coal to a black liquid product which never appeared in cultures unless fungi and coal were present together. Apparently, lignite coal can serve as the principal substrate for the growth of the fungi. Infrared analyses of the liquid products of lignite degradation showed both similarities to and differences from the original lignite.     

An improved approach for transformation of plant cells by microinjection: molecular and genetic analysis

A new culture method for the injection of tobacco mesophyll protoplasts has been established. The protoplasts are embedded in a thin layer of alginate and are nourished from the medium in the underlying basislayer. In the alginate layer the protoplasts regenerate to calli at a frequency of up to 80%. Embedded protoplasts can be selected either with 50 mg l⁻¹ kanamycin or 5 mg l⁻¹ paromomycin. Single resistant cells can be recovered from about 10 000 sensitive cells in one alginate layer. Injection of theneo gene (coding for neomycin phosphotransferase II) into protoplast derived single cells in the alginate layer results in kanamycin resistant colonies that can be regenerated to mature plants. These plants express the neomycin phosphotransferase as shown by enzyme activity assay. The integration of the transgene into the plant genome could be proved by Southern hybridization to high molecular weight DNA. With this culture method 100 cells can be injected per hour. Transformation frequencies range from 2 to 20%. In crossing experiments, it was shown that the foreign gene is transmitted to the next generation in a Mendelian fashion.     

Allosteric characteristics of GTP cyclohydrolase I from Escherichia coli.

The kinetic and regulatory properties of GTP cyclohydrolase I were investigated using an improved enzyme assay and direct determination of the product, dihydroneopterin triphosphate. The enzyme was purified from Escherichia coli to absolute homogeneity as demonstrated by N-terminal sequencing of up to 50 amino acid residues. A 30-residue internal fragment showed 42% similarity with rat liver GTP cyclohydrolase I. The enzyme did not obey Michaelis-Menten kinetics or show a sigmoid reaction curve. The substrate saturation kinetics were found to be slow with low response to minor changes in GTP concentrations. GTP cyclohydrolase I has a relatively high apparent Km. The values are slightly different for enzyme purified by GTP-agarose (100 microM) and UTP-agarose (110 microM). Low turnover numbers of 12/min and 19/min were calculated for the respective enzyme preparations. GTP-cyclohydrolase-I activity was modulated in Vmax by K+, divalent cations, UTP and tetrahydrobiopterin. Divalent cations, such as Mg2+, had an activating effect with an optimum at 8 mM Mg2+. A different catalytic function and formation of a new, unidentified product by GTP cyclohydrolase I was observed in the presence of Ca2+. In the presence of 1 mM EDTA and Mg2+, GTP-cyclohydrolase-I activity was strongly inhibited by chelate complexes. UTP proved not to be a competitive inhibitor, but a positive modulator. The inhibition by chelate complexes was totally abolished by UTP. Tetrahydrobiopterin showed an inhibitory effect, with 50% inhibition at 100 microM tetrahydrobiopterin. UTP was able to reduce the inhibition by tetrahydrobiopterin. Using monoclonal antibody 1F11 (related to the GTP-binding site), and monoclonal antibody NS7 (mimicking tetrahydrobiopterin), different binding sites were demonstrated for GTP and tetrahydrobiopterin on each enzyme subunit. Western-blot competition analysis revealed a UTP-binding site different from the binding sites of GTP and tetrahydrobiopterin. Based on the kinetic behaviour and the kind of modulations observed we defined GTP cyclohydrolase I as an M-class allosteric enzyme.     

Steuerung und potentielle Evolutionsgeschwindigkeit des obligaten Teilzieherverhaltens: Ergebnisse eines Zweiweg-Selektionsexperiments mit der M?nchsgrasmücke (Sylvia atricapilla)

Zusammenfassung 1. In Südfrankreich lebt eine obligat teilziehende Population der Mönchsgrasmücke (Sylvia atricapilla) mit rund drei Vierteln Zugvögeln und einem Viertel Standvögeln. Das Teilzugverhalten dieser Population wird in beträchtlichem Umfang von genetischen Faktoren gesteuert (Berthold &Querner 1982). Der starke Selektionserfolg eines Zweiweg-Selektionsexperiments bis zur F₁-Generation ließ eine hohe potentielle Evolutionsgeschwindigkeit des obligaten Teilzieherverhaltens vermuten (2). Wir haben das Evolutionspotential in einem Langzeit-Selektionsversuch experimentell untersucht.2. Wir zogen insgesamt 267 Individuen der freilebenden Mönchsgrasmückenpopulation in fünf Gruppen von Hand auf, und alle fünf Stichproben zeigten entsprechende Verhältnisse von Ziehern und Nichtziehern. Wir konnten das Langzeit-Zweiweg-Selektionsexperiment bis zu einer F₆-Generation durchführen und konnten dabei insgesamt 455 Mönchsgrasmücken in Volieren züchten.3. Das Zweiweg-Selektionsexperiment zeigt, daß die untersuchte teilziehende Mönchsgrasmückenpopulation bereits nach drei Generationen eine ausschließlich ziehende und nach vier bis sechs Generationen eine fast nicht mehr ziehende Population werden kann.4. Das Zweiweg-Selektionsexperiment zeigt starken Selektionserfolg und hohe Heritabilitätswerte (ca. 0,6–1), die deutlich höher liegen als bisher für das Verhalten von (meist domestizierten) Wirbeltieren angegeben. Demnach sind genetische Faktoren für die Steuerung dieses obligaten Teilzugs sehr bedeutsam und wahrscheinlich dominant über Umwelteinflüsse.5. Die Versuchsergebnisse lassen im Falle von schnellen Umweltveränderungen und der Wirkung von starken Selektionsfaktoren ein enorm großes Evolutionspotential des obligaten Teilzieherverhaltens annehmen und damit allgemein eine schnelle Mikroevolution der Ortsbewegung von Vögeln. Sollten die von Meteorologen prophezeiten Klimaänderungen in Bälde eintreten (Erwärmung aufgrund von Verunreinigungen der Atmosphäre durch den Menschen, Treibhauseffekt), dann könnten viele obligate Teilzieher auf genetischer Basis rasch zu Standvögeln werden und wenig ausgeprägte Zieher rasch zu Teilziehern. Unter diesem allgemeinen Anwachsen von Standvögeln könnten ausgeprägte Zugvögel mehr und mehr zu leiden haben.6. Eine genetische Kontrolle obligaten Teilzugs scheint weit verbreitet zu sein, und sie stellt wohl auch für alle Individuen den zuverlässigsten Steuerungsmechanismus dar. Beim fakultativen Teilzug hingegen dürften Umweltfaktoren mehr unmittelbar steuernd wirken.

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Control and evolutionary potential of obligate partial migration: results of a two-way selective breeding experiment with the Blackcap (Sylvia atricapilla)

Summary 1. In southern France breeds an obligate partially migratory population of the Blackcap with about three quarters of migrants and one quarter of residents. Partial migration of this population is considerably controlled by genetic factors (Berthold &Querner 1982). The large selection response of a two-way selective breeding experiment up to F₁-generations suggested a high evolutionary potential of obligate partial migration (2). We have this evolutionary potential experimentally investigated in a long-term selective breeding study.2. We raised by hand a total of 267 individuals of the free-living Blackcap population in five groups, and all five samples showed corresponding ratios of migrants and nonmigrants. We were able to conduct the long-term selective breeding experiment up to a F₆-generation, and thereby a total of 455 Blackcaps could be bred in aviaries.3. The two-way selective breeding experiment shows, that the investigated partially migratory Blackcap population can become completely migratory in just three generations and almost exclusively sedentary in just four to six generations.4. The two-way selective breeding experiment yielded large selection responses and high heritability values (about 0.6–1) which are distinctly higher than those previously reported for behavioural traits of (mostly domesticated) vertebrates. Hence, genetic factors are very important for the control of this obligate partial migration and are presumedly dominant over environmental influences.5. In case of rapid environmental changes and the effect of strong selection factors the experimental results suggest a strikingly high evolutionary potential of obligate partial migration and thus in general a rapid microevolution of bird movements. If climatic changes would occur in the near future as they are predicted by meteorologists (rise in temperature due to man-made pollution of the atmosphere, greenhouse-effect) then many obligate partial migrants could rapidly become sedentary on a genetic basis and many less typical migrants could then considerably suffer from the general increase of residents.6. A genetic control of obligate partial migration appears to be widespread and it also appears to be the most reliable control mechanism for all individuals of such a partially migratory population. Facultative partial migration, however, appears to be controlled more directly by environmental factors.

     

Function-Triggering Antibodies to the Adhesion Molecule L1 Enhance Recovery after Injury of the Adult Mouse Femoral Nerve

L1 is among the few adhesion molecules that favors repair after trauma in the adult central nervous system of vertebrates by promoting neuritogenesis and neuronal survival, among other beneficial features. In the peripheral nervous system, L1 is up-regulated in Schwann cells and regrowing axons after nerve damage, but the functional consequences of this expression remain unclear. Our previous study of L1-deficient mice in a femoral nerve injury model showed an unexpected improved functional recovery, attenuated motoneuronal cell death, and enhanced Schwann cell proliferation, being attributed to the persistent synthesis of neurotrophic factors. On the other hand, transgenic mice over-expressing L1 in neurons led to improved remyelination, but not improved functional recovery. The present study was undertaken to investigate whether the monoclonal L1 antibody 557 that triggers beneficial L1 functions in vitro would trigger these also in femoral nerve repair. We analyzed femoral nerve regeneration in C57BL/6J mice that received this antibody in a hydrogel filled conduit connecting the cut and sutured nerve before its bifurcation, leading to short-term release of antibody by diffusion. Video-based quantitative analysis of motor functions showed improved recovery when compared to mice treated with conduits containing PBS in the hydrogel scaffold, as a vehicle control. This improved recovery was associated with attenuated motoneuron loss, remyelination and improved precision of preferential motor reinnervation. We suggest that function-triggering L1 antibodies applied to the lesion site at the time of injury over a limited time period will not only be beneficial in peripheral, but also central nervous system regeneration.     

A Design Pattern for Decentralised Decision Making

The engineering of large-scale decentralised systems requires sound methodologies to guarantee the attainment of the desired macroscopic system-level behaviour given the microscopic individual-level implementation. While a general-purpose methodology is currently out of reach, specific solutions can be given to broad classes of problems by means of well-conceived design patterns. We propose a design pattern for collective decision making grounded on experimental/theoretical studies of the nest-site selection behaviour observed in honeybee swarms (Apis mellifera). The way in which honeybee swarms arrive at consensus is fairly well-understood at the macroscopic level. We provide formal guidelines for the microscopic implementation of collective decisions to quantitatively match the macroscopic predictions. We discuss implementation strategies based on both homogeneous and heterogeneous multiagent systems, and we provide means to deal with spatial and topological factors that have a bearing on the micro-macro link. Finally, we exploit the design pattern in two case studies that showcase the viability of the approach. Besides engineering, such a design pattern can prove useful for a deeper understanding of decision making in natural systems thanks to the inclusion of individual heterogeneities and spatial factors, which are often disregarded in theoretical modelling.     

Bisulfite compounds as metabolic inhibitors: nonspecific effects on membranes

Bisulfite compounds are shown to be nonspecific inhibitors ofphotosynthetic processes and of ion transport in green tissues.CO₂ fixation and light-dependent transient changes in externalpH are inhibited about 50% by 5x10^–4 M glyoxal-Na-bisulfite.Chloride uptake in the light and in the dark is inhibited tothe same extent at this concentration. At 5x10^–3 M theinhibitor reduces ATP levels in the light and in the dark, andeffects on glycolate oxidase and PEP carboxylase are observed.The extent of inhibition is dependent on time of treatment withglyoxal-Na-bisulfite and freshly prepared NaHSO₃ is equallyas effective as the addition compound. Possible explanations of the bisulfite effects and the relationshipsto SO₂ effects on photosynthesis are discussed. (Received September 1, 1971; )