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'Gelegenheitsdichtung'
(2010)
Background: Leishmania tarentolae, a unicellular eukaryotic protozoan, has been established as a novel host for recombinant protein production in recent years. Current protocols for protein expression in Leishmania are, however, time consuming and require extensive lab work in order to identify well-expressing cell lines. Here we established an alternative protein expression work-flow that employs recently engineered infrared fluorescence protein (IFP) as a suitable and easy-to-handle reporter protein for recombinant protein expression in Leishmania. As model proteins we tested three proteins from the plant Arabidopsis thaliana, including a NAC and a type-B ARR transcription factor. Results: IFP and IFP fusion proteins were expressed in Leishmania and rapidly detected in cells by deconvolution microscopy and in culture by infrared imaging of 96-well microtiter plates using small cell culture volumes (2 μL - 100 μL). Motility, shape and growth of Leishmania cells were not impaired by intracellular accumulation of IFP. In-cell detection of IFP and IFP fusion proteins was straightforward already at the beginning of the expression pipeline and thus allowed early pre-selection of well-expressing Leishmania clones. Furthermore, IFP fusion proteins retained infrared fluorescence after electrophoresis in denaturing SDS-polyacrylamide gels, allowing direct in-gel detection without the need to disassemble cast protein gels. Thus, parameters for scaling up protein production and streamlining purification routes can be easily optimized when employing IFP as reporter. Conclusions: Using IFP as biosensor we devised a protocol for rapid and convenient protein expression in Leishmania tarentolae. Our expression pipeline is superior to previously established methods in that it significantly reduces the hands-on-time and work load required for identifying well-expressing clones, refining protein production parameters and establishing purification protocols. The facile in-cell and in-gel detection tools built on IFP make Leishmania amenable for high-throughput expression of proteins from plant and animal sources.
Background: Cysteine is a component in organic compounds including glutathione that have been implicated in the adaptation of plants to stresses. O-acetylserine (thiol) lyase (OAS-TL) catalyses the final step of cysteine biosynthesis. OAS-TL enzyme isoforms are localised in the cytoplasm, the plastids and mitochondria but the contribution of individual OAS-TL isoforms to plant sulphur metabolism has not yet been fully clarified.
Results: The seedling lethal phenotype of the Arabidopsis onset of leaf death3-1 (old3-1) mutant is due to a point mutation in the OAS-A1 gene, encoding the cytosolic OAS-TL. The mutation causes a single amino acid substitution from Gly(162) to Glu(162), abolishing old3-1 OAS-TL activity in vitro. The old3-1 mutation segregates as a monogenic semidominant trait when backcrossed to its wild type accession Landsberg erecta (Ler-0) and the Di-2 accession. Consistent with its semi-dominant behaviour, wild type Ler-0 plants transformed with the mutated old3-1 gene, displayed the early leaf death phenotype. However, the old3-1 mutation segregates in an 11: 4: 1 (wild type: semi-dominant: mutant) ratio when backcrossed to the Colombia-0 and Wassilewskija accessions. Thus, the early leaf death phenotype depends on two semi-dominant loci. The second locus that determines the old3-1 early leaf death phenotype is referred to as odd-ler (for old3 determinant in the Ler accession) and is located on chromosome 3. The early leaf death phenotype is temperature dependent and is associated with increased expression of defence-response and oxidative-stress marker genes. Independent of the presence of the odd-ler gene, OAS-A1 is involved in maintaining sulphur and thiol levels and is required for resistance against cadmium stress.
Conclusions: The cytosolic OAS-TL is involved in maintaining organic sulphur levels. The old3-1 mutation causes genome-dependent and independent phenotypes and uncovers a novel function for the mutated OAS-TL in cell death regulation.
Afrika hat seinen berechtigten Platz auf der Weltbühne noch nicht gefunden. Ein wichtiger Grund dafür besteht darin, dass afrikanische Regierungen – im Gegensatz zu denen Chinas, Indiens, Brasiliens oder Indonesiens – nicht jene Maßnahmen ergriffen haben, die für das Entstehen eines Mittelstandes aus dem Gemenge von Millionen Armen wichtig sind.
Der Aufsatz befasst sich mit der Frage, was man aus der internationalen Forschung darüber weiß, welcher Unterricht und welches Lehrerverhalten Auswirkungen auf die Lernergebnisse der Schüler/innen hat und welche Möglichkeiten es gibt, solches Verhalten zu verstärken. Berücksichtigt werden nur Forschungsergebnisse, die hohen methodischen Anforderungen genügen, kausale Schlussfolgerungen erlauben und generalisierbar sind. Konsequenzen für die Lehrerbildung werden diskutiert.
Background: For heterogeneous tissues, such as blood, measurements of gene expression are confounded by relative proportions of cell types involved. Conclusions have to rely on estimation of gene expression signals for homogeneous cell populations, e.g. by applying micro-dissection, fluorescence activated cell sorting, or in-silico deconfounding. We studied feasibility and validity of a non-negative matrix decomposition algorithm using experimental gene expression data for blood and sorted cells from the same donor samples. Our objective was to optimize the algorithm regarding detection of differentially expressed genes and to enable its use for classification in the difficult scenario of reversely regulated genes. This would be of importance for the identification of candidate biomarkers in heterogeneous tissues.
Results: Experimental data and simulation studies involving noise parameters estimated from these data revealed that for valid detection of differential gene expression, quantile normalization and use of non-log data are optimal. We demonstrate the feasibility of predicting proportions of constituting cell types from gene expression data of single samples, as a prerequisite for a deconfounding-based classification approach. Classification cross-validation errors with and without using deconfounding results are reported as well as sample-size dependencies. Implementation of the algorithm, simulation and analysis scripts are available.
Conclusions: The deconfounding algorithm without decorrelation using quantile normalization on non-log data is proposed for biomarkers that are difficult to detect, and for cases where confounding by varying proportions of cell types is the suspected reason. In this case, a deconfounding ranking approach can be used as a powerful alternative to, or complement of, other statistical learning approaches to define candidate biomarkers for molecular diagnosis and prediction in biomedicine, in realistically noisy conditions and with moderate sample sizes.
Multi-color fluorescence imaging experiments of wave forming Dictyostelium cells have revealed that actin waves separate two domains of the cell cortex that differ in their actin structure and phosphoinositide composition. We propose a bistable model of actin dynamics to account for these experimental observation. The model is based on the simplifying assumption that the actin cytoskeleton is composed of two distinct network types, a dendritic and a bundled network. The two structurally different states that were observed in experiments correspond to the stable fixed points in the bistable regime of this model. Each fixed point is dominated by one of the two network types. The experimentally observed actin waves can be considered as trigger waves that propagate transitions between the two stable fixed points.
Die faulige Luft aus dem tiefsten Mittelalter, denken sich zivilisierte Mitteleuropabewohnende, und sind erstaunt zu erfahren, dass es im Iran heute legal ist, sich von einer Frau in einen Mann umoperieren zu lassen, oder umgekehrt. Noch besser: Die Geschlechtsumwandlung wird von der Krankenkasse bezahlt.
Brodelnder Iran
(2010)
Die letzten Wahlen im Iran waren die Zündschnur am Pulverfass: Der angestaute Protest entlud sich. Wenig organisiert, jedoch mit Kreativität und erstaunlicher Dauer der Opposition. Zugleich gelingt es dem Regime weiterhin, mit Härte gegen oppositionelle Kräfte vorzugehen. So brodelt es bedrohlich im Iran. Afghanistan bleibt hierzulande auf der Tagesordnung. Obamas "neue Strategie" erinnert an Vietnam. Mit ihr wird sich der Druck auf Berlin verstärken, die Zahl der Soldaten am Hindukusch zu erhöhen. Dies erfordert weiterhin eine Debatte um Alternativen.
A large number and wide variety of lake ecosystem models have been developed and published during the past four decades. We identify two challenges for making further progress in this field. One such challenge is to avoid developing more models largely following the concept of others ('reinventing the wheel'). The other challenge is to avoid focusing on only one type of model, while ignoring new and diverse approaches that have become available ('having tunnel vision'). In this paper, we aim at improving the awareness of existing models and knowledge of concurrent approaches in lake ecosystem modelling, without covering all possible model tools and avenues. First, we present a broad variety of modelling approaches. To illustrate these approaches, we give brief descriptions of rather arbitrarily selected sets of specific models. We deal with static models (steady state and regression models), complex dynamic models (CAEDYM, CE-QUAL-W2, Delft 3D-ECO, LakeMab, LakeWeb, MyLake, PCLake, PROTECH, SALMO), structurally dynamic models and minimal dynamic models. We also discuss a group of approaches that could all be classified as individual based: super-individual models (Piscator, Charisma), physiologically structured models, stage-structured models and traitbased models. We briefly mention genetic algorithms, neural networks, Kalman filters and fuzzy logic. Thereafter, we zoom in, as an in-depth example, on the multi-decadal development and application of the lake ecosystem model PCLake and related models (PCLake Metamodel, Lake Shira Model, IPH-TRIM3D-PCLake). In the discussion, we argue that while the historical development of each approach and model is understandable given its 'leading principle', there are many opportunities for combining approaches. We take the point of view that a single 'right' approach does not exist and should not be strived for. Instead, multiple modelling approaches, applied concurrently to a given problem, can help develop an integrative view on the functioning of lake ecosystems. We end with a set of specific recommendations that may be of help in the further development of lake ecosystem models.
Die Frage, ob die CDU-Bundestagsabgeordnete Erika Steinbach, seit 1998 Präsidenten des Bundes der Vertriebenen (BdV), einen Sitz im Stiftungsrat des "Zentrums gegen Vertreibung" einnehmen soll, spaltet die politische Landschaft. Die von der SPD beantragte Aktuelle Stunde des Bundestages am 26. November 2009 machte dies erneut deutlich. Auch die neue Bundesregierung ist in dieser Frage gespalten.
Das Ende der Theokratie?
(2010)
Iran steht vor einer neuen Revolution. Die Wahlen und deren offensichtlicher Betrug haben die Spaltung des Landes zu Tage gefördert. Die theokratische Herrschaft kann sich nur noch durch einen ölgeförderten Klientelismus am Leben erhalten und steht nun dort, wo der Schah vor dreißig Jahren gestanden hat. Nur durch eine kluge Politik, die auch Versöhnung bietet, kann ein friedlicher Machtwechsel herbeigeführt und die Demokratisierung des Landes ermöglicht werden.
Außenpolitik ist von den Werten, dem Selbstverständnis einer Gesellschaft und den ökonomischen Interessen ihrer Volkswirtschaft geprägt. In den ersten beiden Teilen dieses Essays zeigen wir auf, wie Deutschland seine Werte behutsam erneuern sollte und welche Bereiche zur Wahrung seines ökonomischen Wohlstands zentral sind. Auf dieses Fundament setzen wir ein Plädoyer für eine neue außenpolitische Kultur, die Interessen offen debattiert und klar definiert, um sie erfolgreich durchzusetzen.
Der Aufsatz befasst sich mit der Frage, was man aus der internationalen Forschung darüber weiß, welcher Unterricht und welches Lehrerverhalten Auswirkungen auf die Lernergebnisse der Schüler/innen hat und welche Möglichkeiten es gibt, solches Verhalten zu verstärken. Berücksichtigt werden nur Forschungsergebnisse, die hohen methodischen Anforderungen genügen, kausale Schlussfolgerungen erlauben und generalisierbar sind. Konsequenzen für die Lehrerbildung werden diskutiert.
ERP-Architekturen
(2010)
Es geht nicht um "Versagen"!
(2010)
Die sich zunächst um Griechenland rankende Krise wurde zu einer europäischen, die den Euro bedroht. Allenthalben wird die Frage gestellt: Wer ist schuld? In deutschen Groß-Medien wird auf das Zögern der derzeitigen Bundesregierung verwiesen. Der Euro rutschte in die Gefahrenzone. Aber ist der Verweis auf falsche Politik die Antwort auf die Frage nach dem erneuten Aufbrechen der Finanzkrise?
We present an approach that provides automatic or semi-automatic support for evolution and change management in heterogeneous legacy landscapes where (1) legacy heterogeneous, possibly distributed platforms are integrated in a service oriented fashion, (2) the coordination of functionality is provided at the service level, through orchestration, (3) compliance and correctness are provided through policies and business rules, (4) evolution and correctness-by-design are supported by the eXtreme Model Driven Development paradigm (XMDD) offered by the jABC (Margaria and Steffen in Annu. Rev. Commun. 57, 2004)—the model-driven service oriented development platform we use here for integration, design, evolution, and governance. The artifacts are here semantically enriched, so that automatic synthesis plugins can field the vision of Enterprise Physics: knowledge driven business process development for the end user.
We demonstrate this vision along a concrete case study that became over the past three years a benchmark for Semantic Web Service discovery and mediation. We enhance the Mediation Scenario of the Semantic Web Service Challenge along the 2 central evolution paradigms that occur in practice: (a) Platform migration: platform substitution of a legacy system by an ERP system and (b) Backend extension: extension of the legacy Customer Relationship Management (CRM) and Order Management System (OMS) backends via an additional ERP layer.
Models employed in exercise psychology highlight the role of reflective processes for explaining behavior change. However, as discussed in social cognition literature, information-processing models also consider automatic processes (dual-process models). To examine the relevance of automatic processing in exercise psychology, we used a priming task to assess the automatic evaluations of exercise stimuli in physically active sport and exercise majors (n = 32), physically active nonsport majors (n = 31), and inactive students (n = 31). Results showed that physically active students responded faster to positive words after exercise primes, whereas inactive students responded more rapidly to negative words. Priming task reaction times were successfully used to predict reported amounts of exercise in an ordinal regression model. Findings were obtained only with experiential items reflecting negative and positive consequences of exercise. The results illustrate the potential importance of dual-process models in exercise psychology.
Exit Afghanistan
(2010)
Den Krieg um Afghanistan hat der Westen verloren - wieder einmal. Die Frage des Abzugs aus dem Land am Hindukusch bestimmt die außen- und sicherheitspolitische Agenda, nicht nur in Washington D.C. und Berlin. Ziel ist, die gescheiterte Intervention möglichst glimpflich enden zu lassen: "Exit Afghanistan" - aber wie?