In vivo consequences of AML1-ETO fusion protein expression for hematopoiesis

The t(8;21) (q22;q22) translocation fusing the ETO (also known as MTG8) gene on human chromosome 8 with the AML1 (also called Runx1 or CBFα) gene on chromosome 21 is one of the most common genetic aberrations found in acute myeloid leukemia (AML). This chromosomal translocation occurs in 12 % of de novo AML cases and in up to 40 % of the AML-M2 subtype of the French-American-British classification. To date, the in vivo function of aberrant AML1-ETO fusion protein expression has been investigated by several groups. However, in these studies, controversial results were reported and some key issues remain unknown. Importantly, the consequences of aberrant AML1-ETO expression for self-renewing hematopoietic stem cells (HSCs), multipotent hematopoietic progenitors (MPPs) and lineage-restricted precursors are not known. rn The aim of this thesis was to develop a novel experimental AML1-ETO in vivo model that (i) overcomes the current lack of insight into the pre-leukemic condition of t(8;21)-associated AML, (ii) clarifies the in vivo consequences of AML1-ETO for HSCs, MPPs, progenitors and more mature blood cells and (iii) generates an improved mouse model suitable for mirroring the human condition. For this purpose, a conditional tet on/off mouse model expressing the AML1-ETO fusion protein from the ROSA26 (R26) locus was generated. rn Aberrant AML1-ETO activation in compound ROSA26/tetOAML1-ETO (R26/AE) mice caused high rates of mortality, an overall disruption of hematopoietic organs and a profound alteration of hematopoiesis. However, since the generalized activity of the R26 locus did not recapitulate the leukemic condition found in human patients, it was important to restrict AML1-ETO expression to blood cell lineages. Therefore, bone marrow cells from non-induced R26/AE mice were adoptively transplanted into sublethal irradiated RAG2-/- recipient mice. First signs of phenotypical differences between AML1-ETO-expressing and control mice were observed after eight to nine months of transgene induction. AML1-ETO-expressing mice showed profound changes in hematopoietic organs accompanied by manifest extramedullary hematopoiesis. In addition, a block in early erythropoiesis, B- and T-cell maturation was observed and granulopoiesis was significantly enhanced. Most interestingly, conditional activation of AML1-ETO in chimeric mice did not increase HSCs, MPPs, common lymphoid precursors (CLPs), common myeloid progenitors (CMPs) and megakaryocyte-erythrocyte progenitors (MEPs) but promoted the selective amplification of granulocyte-macrophage progenitors (GMPs). rn The results of this thesis provide clear experimental evidence how aberrant AML1-ETO modulates the developmental properties of normal hematopoiesis and establishes for the first time that AML1-ETO does not increase HSCs, MPPs and common lineage-restricted progenitor pools but specifically amplifies GMPs. The here presented mouse model not only clarifies the role of aberrant AML1-ETO for shaping hematopoietic development but in addition has strong implications for future therapeutic strategies and will be an excellent pre-clinical tool for developing and testing new approaches to treat and eventually cure AML.rn

Bayesian space-time data fusion for real-time forecasting and map uncertainty

Environmental computer models are deterministic models devoted to predict several environmental phenomena such as air pollution or meteorological events. Numerical model output is given in terms of averages over grid cells, usually at high spatial and temporal resolution. However, these outputs are often biased with unknown calibration and not equipped with any information about the associated uncertainty. Conversely, data collected at monitoring stations is more accurate since they essentially provide the true levels. Due the leading role played by numerical models, it now important to compare model output with observations. Statistical methods developed to combine numerical model output and station data are usually referred to as data fusion. In this work, we first combine ozone monitoring data with ozone predictions from the Eta-CMAQ air quality model in order to forecast real-time current 8-hour average ozone level defined as the average of the previous four hours, current hour, and predictions for the next three hours. We propose a Bayesian downscaler model based on first differences with a flexible coefficient structure and an efficient computational strategy to fit model parameters. Model validation for the eastern United States shows consequential improvement of our fully inferential approach compared with the current real-time forecasting system. Furthermore, we consider the introduction of temperature data from a weather forecast model into the downscaler, showing improved real-time ozone predictions. Finally, we introduce a hierarchical model to obtain spatially varying uncertainty associated with numerical model output. We show how we can learn about such uncertainty through suitable stochastic data fusion modeling using some external validation data. We illustrate our Bayesian model by providing the uncertainty map associated with a temperature output over the northeastern United States.

Sensor Data Fusion e applicazioni

Un trattato che illustra tutto quello che riguarda il Sensor Data Fusion sin dagli strumenti basilari fino alle applicazioni di specializzate

Flagellin:allergen fusion proteins as novel vaccines for the treatment of severe type I allergies

Over the last decades the prevalence of food allergies has continually increased on a world wide scale. While there are effective treatments available for bee and wasp venom allergic patients, there is currently no established therapy for the treatment of severe food allergies. Aim of the project was to genetically fuse different food allergens with the immune modulating Toll-like receptor 5 (TLR5)-ligand flagellin and to test these constructs for their immune modulatory capacities both in vitro and in vivo. Chicken ovalbumin (Ova) as model antigen, Pru p 3, and Ara h 2 the respective major allergens from peach and peanut were used as allergens. The potential vaccine candidates were characterized by protein biochemical methods (purity, folding, endotoxin contaminations). Moreover, their immune modulating effects on cell culture lines (TLR5-receptor activation) and primary mouse immune cells (myeloid and plasmacytoid dendritic cells) were investigated. Additionally, the prophylactic and therapeutic use of the flagellin Ova fusion protein (rflaA:Ova) were investigated in a mouse model of intestinal allergy. In myeloid dendritic cells (mDC) stimulation with the fusion proteins led to a strong cell activation and cytokine secretion. Here, the fusion proteins proved to be a much stronger stimulus than the equimolar amount of both proteins provided alone or as a mixture. Noteworthy, stimulation with rflaA:Ova induced the secretion of the anti-inflammatory cytokine IL-10 from mDC. In co-culture experiments this IL-10 secretion suppressed the Ova-induced secretion of Th1 and Th2 cytokines from Ova-specific CD4 T cells. Using MyD88-deficient mDC this repression of cytokine secretion was shown to be TLR-dependent. Finally, the potency of the rflaA:Ova construct was investigated in a mouse model of Ova-induced intestinal allergy. In a prophylactic vaccination approach rflaA:Ova was shown to prevent the establishment of the intestinal allergy and all associated symptoms (weight loss, temperature drop, soft faeces). This fusion protein-mediated protection was accompanied by a reduced T cell activation, and reduced Th2 cytokines in intestinal homogenates. These effects were paralleled by a strong induction of Ova-specific IgG2a antibodies in rflaA:Ova-vaccinated sera, while Ova-specific IgE antibody production was significantly reduced. Therapeutic vaccination with rflaA:Ova reduced allergic symptoms and T cell activation but did not influence weight loss and antibody production. In all in vivo experiments vaccination with both proteins either provided alone or as a mixture did not have comparable effects. Future experiments aim at elucidating the mechanism and further optimization of the therapeutic vaccination approach. The results presented in this thesis demonstrate, that fusion proteins containing flagellin have strong immune modulatory capacities both in vitro and in vivo. Therefore, such constructs are promising vaccine candidates for the therapy of type I allergies.

A new information fusion method for land-use classification using high resolution satellite imagery

Satellite image classification involves designing and developing efficient image classifiers. With satellite image data and image analysis methods multiplying rapidly, selecting the right mix of data sources and data analysis approaches has become critical to the generation of quality land-use maps. In this study, a new postprocessing information fusion algorithm for the extraction and representation of land-use information based on high-resolution satellite imagery is presented. This approach can produce land-use maps with sharp interregional boundaries and homogeneous regions. The proposed approach is conducted in five steps. First, a GIS layer - ATKIS data - was used to generate two coarse homogeneous regions, i.e. urban and rural areas. Second, a thematic (class) map was generated by use of a hybrid spectral classifier combining Gaussian Maximum Likelihood algorithm (GML) and ISODATA classifier. Third, a probabilistic relaxation algorithm was performed on the thematic map, resulting in a smoothed thematic map. Fourth, edge detection and edge thinning techniques were used to generate a contour map with pixel-width interclass boundaries. Fifth, the contour map was superimposed on the thematic map by use of a region-growing algorithm with the contour map and the smoothed thematic map as two constraints. For the operation of the proposed method, a software package is developed using programming language C. This software package comprises the GML algorithm, a probabilistic relaxation algorithm, TBL edge detector, an edge thresholding algorithm, a fast parallel thinning algorithm, and a region-growing information fusion algorithm. The county of Landau of the State Rheinland-Pfalz, Germany was selected as a test site. The high-resolution IRS-1C imagery was used as the principal input data.

Il concetto del "ma" e la musica di Tōru Takemitsu: traduzione di "The concept of 'ma' and the music of Takemitsu" di Jonathan L. Chenette

Lo scopo di questa tesi è di esplorare l'importanza del concetto giapponese del "ma" nella musica tradizionale, in particolare in quella del compositore Tōru Takemitsu, tramite la traduzione del saggio "The concept of 'ma' and the music of Tōru Takemitsu" (Jonathan L. Chenette, 1982) dall'inglese all'italiano. L'elaborato partirà da un'introduzione generale sul concetto del "ma" nella mentalità giapponese, per proseguire con la biografia di Tōru Takemitsu e una panoramica dei motivi che hanno portato a scegliere la traduzione del saggio di Chenette. Dopo la traduzione in sé e per sé e il commento della stessa, sarà anche fornito in appendice un glossario inglese-italiano della terminologia musicale utilizzata dall'autore all'interno del saggio.

Tradurre la popular music. Il caso di studio Ministri per il mercato estero

La presente tesi si concentra sulla traduzione musicale, focalizzandosi in particolare sulla traduzione di canzoni popular. L’obiettivo è creare dei testi di arrivo cantabili in lingua inglese a partire da originali del gruppo musicale italiano Ministri, così da rendere esportabile il loro prodotto oltre i confini del mercato discografico nazionale. La struttura della tesi è composta da quattro capitoli e due appendici. Nel primo capitolo si individua la cornice all’interno del quale si muove il caso di studio selezionato e si inserisce l’attività di traduzione. Si analizzano nello specifico alcune caratteristiche essenziali della popular music, i suoi testi, gli attori e gli intermediari coinvolti. Nel secondo capitolo si analizza il caso di studio, passando in rassegna rapidamente la biografia, l’estetica e la modalità di comunicazione del gruppo, per poi approfondirne in maniera particolare una parte della produzione, con speciale enfasi sui testi verbali che vanno a costituire il materiale oggetto di traduzione. Nel terzo capitolo si individua nella teoria dello scopo la base teorica per la metodologia specifica della traduzione di canzoni. Quest’ultima rientra nella traduzione musicale, appartenente alle categorie di traduzione vincolata e multimediale. Nel quarto capitolo viene esposto l’obiettivo della traduzione e la metodologia specifica utilizzata per portarlo a termine, passando poi a un esteso commento delle problematiche traduttive individuate e delle strategie utilizzate per risolverle, apportando alcuni esempi per ogni caso. Infine, nella prima appendice vengono riportati integralmente i testi originali con le relative proposte di traduzione, mentre la seconda contiene un piccolo contributo di Federico Dragogna, chitarrista e principale autore dei Ministri, sotto forma di breve intervista.

A novel fusion toxin derived from an EpCAM-specific designed ankyrin repeat protein has potent antitumor activity

Designed ankyrin repeat proteins (DARPins) hold great promise as a new class of binding molecules to overcome the limitations of antibodies for biomedical applications. Here, we assessed the potential of an epithelial cell adhesion molecule (EpCAM)-specific DARPin (Ec4) for tumor targeting as a fusion toxin with Pseudomonas aeruginosa exotoxin A.