@article{UeceylerTopuzoğluSchiesseretal.2011, author = {{\"U}{\c{c}}eyler, Nurcan and Topuzoğlu, Teng{\"u} and Schießer, Peter and Hahnenkamp, Saskia and Sommer, Claudia}, title = {IL-4 Deficiency Is Associated with Mechanical Hypersensitivity in Mice}, series = {PLoS One}, volume = {6}, journal = {PLoS One}, number = {12}, doi = {10.1371/journal.pone.0028205}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-137924}, pages = {e28205}, year = {2011}, abstract = {Interleukin-4 (IL-4) is an anti-inflammatory and analgesic cytokine that induces opioid receptor transcription. We investigated IL-4 knockout (ko) mice to characterize their pain behavior before and after chronic constriction injury (CCI) of the sciatic nerve as a model for neuropathic pain. We investigated opioid responsivity and measured cytokine and opioid receptor gene expression in the peripheral and central nervous system (PNS, CNS) of IL-4 ko mice in comparison with wildtype (wt) mice. Na{\"i}ve IL-4 ko mice displayed tactile allodynia (wt: 0.45 g; ko: 0.18 g; p<0.001), while responses to heat and cold stimuli and to muscle pressure were not different. No compensatory changes in the gene expression of tumor necrosis factor-alpha (TNF), IL-1β, IL-10, and IL-13 were found in the PNS and CNS of na{\"i}ve IL-4 ko mice. However, IL-1β gene expression was stronger in the sciatic nerve of IL-4 ko mice (p<0.001) 28 days after CCI and only IL-4 ko mice had elevated IL-10 gene expression (p = 0.014). Remarkably, CCI induced TNF (p<0.01), IL-1β (p<0.05), IL-10 (p<0.05), and IL-13 (p<0.001) gene expression exclusively in the ipsilateral spinal cord of IL-4 ko mice. The compensatory overexpression of the anti-inflammatory and analgesic cytokines IL-10 and IL-13 in the spinal cord of IL-4 ko mice may explain the lack of genotype differences for pain behavior after CCI. Additionally, CCI induced gene expression of μ, κ, and δ opioid receptors in the contralateral cortex and thalamus of IL-4 ko mice, paralleled by fast onset of morphine analgesia, but not in wt mice. We conclude that a lack of IL-4 leads to mechanical sensitivity; the compensatory hyperexpression of analgesic cytokines and opioid receptors after CCI, in turn, protects IL-4 ko mice from enhanced pain behavior after nerve lesion.}, language = {en} } @article{UeceylerHaeuserSommer2011, author = {{\"U}ceyler, Nurcan and H{\"a}user, Winfried and Sommer, Claudia}, title = {Systematic review with meta-analysis: Cytokines in fibromyalgia syndrome}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-69189}, year = {2011}, abstract = {Background: To perform a systematic review and meta-analysis on cytokine levels in patients with fibromyalgia syndrome (FMS). Methods: Through December 2010 we systematically reviewed the databases PubMed, MEDLINE, and PsycINFO and screened the reference lists of 22 review articles for suitable original articles. Original articles investigating cytokines in patients with FMS were included. Data were extracted by two independent authors. Differences of the cytokine levels of FMS patients and controls were summarized by standardized mean differences (SMD) using a random effects model. Study quality was assessed applying methodological scores: modified Center of Evidence Based Medicine, Newcastle-Ottawa-Scale, and W{\"u}rzburg Methodological Quality Score. Results: Twenty-five articles were included investigating 1255 FMS patients and 800 healthy controls. Data of 13/25 studies entered meta-analysis. The overall methodological quality of studies was low. The results of the majority of studies were not comparable because methods, investigated material, and investigated target cytokines differed. Systematic review of the selected 25 articles revealed that FMS patients had higher serum levels of interleukin (IL)-1 receptor antagonist, IL-6, and IL-8, and higher plasma levels of IL-8. Meta-analysis of eligible studies showed that FMS patients had higher plasma IL-6 levels compared to controls (SMD = -0.34 [-0.64, -0.03] 95\% CI; p = 0.03). The majority of investigated cytokines were not different between patients and controls. Conclusions: The pathophysiological role of cytokines in FMS is still unclear. Studies of higher quality and with higher numbers of subjects are needed.}, subject = {Fibromyalgie}, language = {en} } @phdthesis{OenalHartmann2011, author = {{\"O}nal-Hartmann, Cigdem}, title = {Emotional Modulation of Motor Memory Formation}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-64838}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2011}, abstract = {Hintergr{\"u}nde: Wie eine Vielzahl von Studien belegt, kann das explizite Ged{\"a}chtnis, das die bewusste Erinnerung an enkodierte Informationen beinhaltet, durch Emotionen beeinflusst werden, und zwar {\"u}ber den Einfluss auf verschiedene Verarbeitungsebenen (Enkodierung, Konsolidierung, Abruf usw.). Bisher wenig untersucht ist, ob und wie Emotionen Vorg{\"a}nge der motorischen Ged{\"a}chtnisbildung, die nicht auf bewusster Erinnerung beruhen und sich stattdessen durch Ver{\"a}nderungen im Verhalten darstellen, modulieren. Experiment 1: Das Ziel des ersten Experimentes war es, den Einfluss von Emotionen auf motorisches Lernen zu untersuchen. Vier Gruppen von Probanden mussten in einer motorischen Lernaufgabe schnelle, seitliche Bewegungen mit dem Daumen ausf{\"u}hren. W{\"a}hrend dieser Aufgabe h{\"o}rten die Probanden emotionale Kl{\"a}nge, die in Valenz und Arousal variierten: 1. Valenz negativ/ Arousal niedrig (V-/A-), 2. Valenz negativ/ Arousal hoch (V-/A+), 3. Valenz positiv/ Arousal niedrig (V+/A-), 4. Valenz positiv/ Arousal hoch (V+/A+). Die deskriptive Analyse aller Daten sprach f{\"u}r beste Ergebnisse f{\"u}r das motorische Lernen in der Bedingung V-/A-, aber die Unterschiede zwischen den Bedingungen waren nicht signifikant. Die Interaktion zwischen Valenz und Arousal emotionaler T{\"o}ne scheint demnach motorische Enkodierungsprozesse zu modulieren, jedoch m{\"u}ssen zuk{\"u}nftige Studien mit unterschiedlichen emotionalen Stimuli die Annahme weiter untersuchen, dass negative Stimuli mit niedrigem Arousal w{\"a}hrend der Enkodierung einen f{\"o}rdernden Effekt auf das motorische Kurzzeitged{\"a}chtnis haben. Experiment 2: Die Absicht des zweiten Experimentes war es, die Auswirkungen emotionaler Interferenzen auf die Konsolidierung beim Sequenzlernen zu untersuchen. Sechs Gruppen von Probanden trainierten zuerst in getrennten Sitzungen eine SRTT-Aufgabe (serial reaction time task). Um die Konsolidierung der neu erlernten Fertigkeit zu modulieren, wurden die Probanden nach dem Training einer von drei unterschiedlichen Klassen emotionaler Stimuli (positiv, negativ oder neutral) ausgesetzt. Diese bestanden aus einem Set emotionaler Bilder, die mit emotional kongruenten Musikst{\"u}cken oder neutralen Kl{\"a}ngen kombiniert waren. Bei den Probandengruppen wurde die emotionale Interferenz nach zwei unterschiedlichen Zeitintervallen realisiert, entweder direkt nach der Trainingssitzung oder sechs Stunden sp{\"a}ter. 72 Stunden nach der Trainingssitzung wurde jede Gruppe erneut mit der SRTT-Aufgabe getestet. Die Leistung in diesem Nachtest wurde mittels Reaktionszeit und Genauigkeit bei der Ausf{\"u}hrung der Zielsequenz analysiert. Die emotionale Interferenz beeinflusste weder die Nachtestergebnisse f{\"u}r die Reaktionszeit noch die f{\"u}r die Genauigkeit. Allerdings konnte eine Steigerung der expliziten Sequenzerkennung durch erregende negative Stimuli festgestellt werden, wenn diese direkt nach der ersten Trainingseinheit (0h) dargeboten wurden. Diese Ergebnisse lassen vermuten, dass die Konsolidierung der expliziten Aspekte prozeduralen Lernens in einer st{\"a}rkeren Wechselwirkung mit emotionalen Interferenzen stehen k{\"o}nnte als die der impliziten Aspekte. Die Konsolidierung unterschiedlicher Ebenen des Fertigkeitserwerbs k{\"o}nnte demnach von unterschiedlichen Mechanismen gesteuert werden. Da Performanz und explizites Sequenzerkennen nicht korrelierten, vermuten wir, dass implizite und explizite Modalit{\"a}ten bei der Durchf{\"u}hrung der SRTT-Aufgabe nicht komplement{\"a}r sind. Experiment 3: Es sollte untersucht werden, ob es eine Pr{\"a}ferenz der linken Gehirnhemisph{\"a}re bei der Kontrolle von Flexionsreaktionen auf positive Stimuli gibt und der rechten Hemisph{\"a}re bei der Kontrolle von Extensionsreaktionen auf negative Stimuli. Zu diesem Zweck sollten rechtsh{\"a}ndige Probanden einen Joystick zu sich ziehen oder von sich weg dr{\"u}cken, nachdem sie einen positiven oder negativen Stimulus in ihrem linken oder rechten Gesichtsfeld gesehen hatten. Die Flexionsreaktionen waren bei positiven Stimuli schneller, Extensionsreaktion hingegen bei negativen Stimuli. Insgesamt war die Performanz am schnellsten, wenn die emotionalen Stimuli im linken Gesichtsfeld pr{\"a}sentiert wurden. Dieser Vorrang der rechten Gehirnhemisph{\"a}re war besonders deutlich f{\"u}r negative Stimuli, wohingegen die Reaktionszeiten auf positive Bilder keine hemisph{\"a}rische Differenzierung zeigten. Wir konnten keine Interaktion zwischen Gesichtsfeld und Reaktionstyp belegen, auch fand sich keine Dreifachinteraktion zwischen Valenz, Gesichtsfeld und Reaktionstyp. In unserem experimentellen Kontext scheint die Interaktion zwischen Valenz und Gesichtsfeld st{\"a}rker zu sein als die Interaktion zwischen Valenz und motorischem Verhalten. Auf Grund dieser Ergebnisse vermuten wir, dass unter gewissen Bedingungen eine Hierarchisierung der asymmetrischen Muster Vorrang hat, die m{\"o}glicherweise andere vorhandene Asymmetrien maskieren k{\"o}nnte.}, subject = {Motorisches Lernen}, language = {en} } @phdthesis{ZelmanFemiak2011, author = {Zelman-Femiak, Monika}, title = {Single Particle Tracking ; Membrane Receptor Dynamics}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-65420}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2011}, abstract = {Single-molecule microscopy is one of the decisive methodologies that allows one to clarify cellular signaling in both spatial and temporal dimentions by tracking with nanometer precision the diffusion of individual microscopic particles coupled to relevant biological molecules. Trajectory analysis not only enables determination of the mechanisms that drive and constrain the particles motion but also to reveal crucial information about the molecule interaction, mobility, stoichiometry, all existing subpopulations and unique functions of particular molecules. Efficacy of this technique depends on two problematic issues the usage of the proper fluorophore and the type of biochemical attachment of the fluorophore to a biomolecule. The goal of this study was to evolve a highly specific labeling method suitable for single molecule tracking, internalization and trafficking studies that would attain a calculable 1:1 fluorophore-to-receptor stoichiometry. A covalent attachment of quantum dots to transmembrane receptors was successfully achieved with a techinque that amalgamates acyl carrier protein (ACP) system as a comparatively small linker and coenzyme A (CoA)-functionalized quantum dots. The necessity of optimization of the quantum dot usage for more precise calculation of the membrane protein stoichiometries in larger assemblies led to the further study in which methods maximizing the number of signals and the tracking times of diverse QD types were examined. Next, the optimized techniques were applied to analyze behavior of interleukin-5 β-common chain receptor (IL-5Rβc) receptors that are endogenously expressed at low level on living differentiated eosinophil-like HL-60 cells. Obtained data disclosed that perused receptors form stable and higher order oligomers. Additionally, the mobility analysis based on increased in number (>10\%) uninterrupted 1000-step trajectories revealed two patterns of confined motion. Thereupon methods were developed that allow both, determination of stoichiometries of cell surface protein complexes and the acquisition of long trajectories for mobility analysis. Sequentially, the aforementioned methods were used to scrutinize on the mobility, internalization and recycling dynamics characterization of a G protein-coupled receptor (GPCRs), the parathyroid hormone receptor (PTHR1) and several bone morphogenetic proteins (BMPs), a member of the TGF-beta superfamily of receptors. These receptors are two important representatives of two varied membrane receptor classes. BMPs activate SMAD- and non-SMAD pathways and as members of the transforming growth factor β (TGF-β) superfamily are entailed in the regulation of proliferation, differentiation, chemotaxis, and apoptosis. For effective ligand induced and ligand independent signaling, two types of transmembrane serine/threonine kinases, BMP type I and type II receptors (BMPRI and BMPRII, respectively) are engaged. Apparently, the lateral mobility profiles of BMPRI and BMPRII receptors differ markedly, which determinate specificity of the signal. Non-SMAD signaling and subsequent osteoblastic differentiation of precursor cells particularly necessitate the confinement of the BMP type I receptor, resulting in the conclusion that receptor lateral mobility is a dominative mechanism to modulate SMAD versus non-SMAD signaling during differentiation. Confined motion was also predominantly observed in the studies devoted to, entailed in the regulation of calcium homeostasis and in bone remodeling, the parathyroid hormone receptor (PTHR1), in which stimulation with five peptide ligands, specific fragments of PTH: hPTH(1-34), hPTHrP(107-111)NH2; PTH(1-14); PTH(1-28) G1R19, bPTH(3-34), first four belonging to PTH agonist group and the last to the antagonist one, were tested in the wide concentration range on living COS-1 and AD293 cells. Next to the mobility, defining the internalization and recycling rates of the PTHR1 receptor maintained in this investigation one of the crucial questions. Internalization, in general, allows to diminish the magnitude of the receptor-mediated G protein signals (desensitization), receptor resensitization via recycling, degradation (down-regulation), and coupling to other signaling pathways (e.g. MAP kinases). Determinants of the internalization process are one of the most addressed in recent studies as key factors for clearer understanding of the process and linking it with biological responses evoked by the signal transduction. The internalization of the PTH-receptor complex occurs via the clathrin-coated pit pathway involving β-arrestin2 and is initiated through the agonist occupancy of the PTHR1 leading to activation of adenylyl cyclase (via Gs), and phosphatidylinositol-specific phospholipase Cβ (via Gq). Taken together, this work embodies complex study of the interleukin-5 β-common chain receptor (IL-5Rβc) receptors, bone morphogenetic proteins (BMPs) and the parathyroid hormone receptor with the application of single-molecule microscopy with the newly attained ACP-quantum dot labeling method and standard techniques.}, subject = {Einzelmolek{\"u}lmikroskopie}, language = {en} } @article{ZanuccoGoetzPotapenkoetal.2011, author = {Zanucco, Emanuele and G{\"o}tz, Rudolf and Potapenko, Tamara and Carraretto, Irene and Ceteci, Semra and Ceteci, Fatih and Seeger, Werner and Savai, Rajkumar and Rapp, Ulf R.}, title = {Expression of B-RAF V600E in Type II Pneumocytes Causes Abnormalities in Alveolar Formation, Airspace Enlargement and Tumor Formation in Mice}, series = {PLOS ONE}, volume = {6}, journal = {PLOS ONE}, number = {12}, doi = {10.1371/journal.pone.0029093}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-137061}, pages = {e29093}, year = {2011}, abstract = {Growth factor induced signaling cascades are key regulatory elements in tissue development, maintenance and regeneration. Perturbations of these cascades have severe consequences, leading to developmental disorders and neoplastic diseases. As a major function in signal transduction, activating mutations in RAF family kinases are the cause of human tumorigenesis, where B-RAF V600E has been identified as the prevalent mutant. In order to address the oncogenic function of B-RAF V600E, we have generated transgenic mice expressing the activated oncogene specifically in lung alveolar epithelial type II cells. Constitutive expression of B-RAF V600E caused abnormalities in alveolar epithelium formation that led to airspace enlargements. These lung lesions showed signs of tissue remodeling and were often associated with chronic inflammation and low incidence of lung tumors. The inflammatory cell infiltration did not precede the formation of the lung lesions but was rather accompanied with late tumor development. These data support a model where the continuous regenerative process initiated by oncogenic B-RAF-driven alveolar disruption provides a tumor-promoting environment associated with chronic inflammation.}, language = {en} } @phdthesis{Zanucco2011, author = {Zanucco, Emanuele}, title = {Role of oncogenic and wild type B-RAF in mouse lung tumor models}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-69603}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2011}, abstract = {Von Wachstumsfaktoren regulierte Signalkaskaden sind Schl{\"u}sselelemente in der Gewebeentwicklung und Geweberegeneration. Eine Deregulation dieser Kaskaden f{\"u}hrt zu Entwicklungsst{\"o}rungen und neoplastischen Krankheiten. F{\"u}r viele humane Krebsformen sind aktivierende Mutationen der Kinasen der RAF Familie verantwortlich. Das erste Projekt dieser Doktorarbeit fokussiert auf der Rolle des B-RAF V600E, welches als eine der am h{\"a}ufigsten vorkommenden Mutantionen in humanen Krebszellen identifiziert worden ist. Um die onkogene Funktion des B-RAF V600E zu untersuchen, haben wir transgene Mauslinien hergestellt, welche das aktivierte Onkogen spezifisch in alveolaren Lungenepithelzellen des Typ II exprimieren. Konstitutive Expression des B-RAF V600E f{\"u}hrte zu einer abnormen alveolaren Epithelzellbildung und zu Emphysem-{\"a}hnlichen L{\"a}sionen. Diese L{\"a}sionen wiesen Zeichen einer Gewebsumstrukturierung auf, oft in Assoziation mit chronischer Inflammation und geringer Inzidenz von Lungentumoren. Die Infiltration der entz{\"u}ndlichen Zellen erfolgte erst nach der Entstehung von Emphysem-{\"a}hnlichen L{\"a}sionen und k{\"o}nnte zur sp{\"a}teren Tumorbildung beigetragen haben. Diese Ergebnisse unterst{\"u}tzen ein Modell, in welchem der kontinuierliche regenerative Prozess eine tumorf{\"o}rdernde Umgebung schafft. Dabei induziert die Aktivit{\"a}t des onkogenen B-RAF eine alveolare St{\"o}rung, welche urs{\"a}chlich verantwortlich ist f{\"u}r den kontinuierlichen regenerativen Prozess. Das zweite Projekt fokussiert auf die Rolle von endogenem (wildtypischen) B-RAF in einem durch onkogenes C-RAF induzierten Maus Lungentumormodell. F{\"u}r unsere Untersuchungen haben wir eine Mauslinie geschaffen, in welcher B-RAF in den C-RAF Lungentumoren konditionell eliminiert werden kann. Eine konditionelle Eliminierung des B-RAF hat die Entstehung von Lungentumoren nicht blockiert, aber zu reduziertem Tumorwachstum gef{\"u}hrt. Dieses reduzierte Tumorwachstum konnte auf eine reduzierte Zellproliferation zur{\"u}ckgef{\"u}hrt werden. Außerdem konnten wir durch die B-RAF Elimination eine Reduktion der Intensit{\"a}t der mitogenen Signalkaskade beobachten. Insgesamt deuten die Ergebnisse darauf hin, dass das onkogene Potential von C-RAF in vivo unabh{\"a}ngig von B-RAF ist und eine Kooperation von B-RAF und C-RAF jedoch f{\"u}r die vollst{\"a}ndige Aktivierung der mitogenen Signalkaskade wichtig ist.}, subject = {Lungenkrebs}, language = {en} } @phdthesis{ZamaniPedram2011, author = {Zamani Pedram, Masoud}, title = {Source, facies, and sedimentary environments of the Middle to Upper Jurassic strata in the Kerman and Tabas areas, east-central Iran}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-56758}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2011}, abstract = {The present study concerned mainly on the source, facies, and sedimentary environments of the Middle to Upper Jurassic strata in the Kerman and Tabas areas, east-central Iran. The composition of sandstones, and heavy mineral analysis point to pre-existing sedimentary, low, middle to upper rank metamorphic, and plutonic rocks of the Kalmard, Posht-e-Badam, Bayazeh, and Zarand-Kerman areas as the source rocks. According to the diagram of WELTJE et al. (1998), most samples from the Middle-Upper Jurassic rocks suggest a moderate to high elevation of the source area, and indicate a semi-arid and mediterranean to sub-humid climate. In the Qt-F-L ternary diagrams of DICKINSON et al. (1983), most point counting data from the Lower Siliciclastic Member and the top of the Hojedk Formation plot in the recycled orogen (Quartzose recycled) area of the diagram. The sandstones in this area can be interpreted as being derived from the Mid-Cimmerian Movements. Sixteen different types of siliciclastic-carbonate, and evaporatic sedimentary environments have been recognized. Thirty-nine macroinvertebrate taxa have been identified. Ten ichnotaxa have been taxonomically described from the Middle to Upper Jurassic rocks. Quite likely, before rotation of CEIM which were associated with counterclockwise block-rotation, equivalent rocks of the Bidou Formation occurred along the tectonic zone between the Yazd and the Tabas blocks (probably during the Middle Jurassic to Lower Cretaceous). However, from the Cretaceous onwards, most of the Bidou Formation has been removed by a combination of strike-slip and reverse movements of the Kashmar-Kerman tectonic zone. Roughly, these block-rotation movements occurred after the Cretaceous. During the Middle to Upper Jurassic, the tectonic activities were vertical movements producing the sedimentary pattern in the CEIM.}, subject = {Kerman }, language = {en} } @article{YinBrocherFischeretal.2011, author = {Yin, Jun and Brocher, Jan and Fischer, Utz and Winkler, Christoph}, title = {Mutant Prpf31 causes pre-mRNA splicing defects and rod photoreceptor cell degeneration in a zebrafish model for Retinitis pigmentosa}, series = {Molecular neurodegeneration}, volume = {6}, journal = {Molecular neurodegeneration}, number = {56}, doi = {10.1186/1750-1326-6-56}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-141090}, pages = {1-17}, year = {2011}, abstract = {Background: Retinitis pigmentosa (RP) is an inherited eye disease characterized by the progressive degeneration of rod photoreceptor cells. Mutations in pre-mRNA splicing factors including PRPF31 have been identified as cause for RP, raising the question how mutations in general factors lead to tissue specific defects. Results: We have recently shown that the zebrafish serves as an excellent model allowing the recapitulation of key events of RP. Here we use this model to investigate two pathogenic mutations in PRPF31, SP117 and AD5, causing the autosomal dominant form of RP. We show that SP117 leads to an unstable protein that is mislocalized to the rod cytoplasm. Importantly, its overexpression does not result in photoreceptor degeneration suggesting haploinsufficiency as the underlying cause in human RP patients carrying SP117. In contrast, overexpression of AD5 results in embryonic lethality, which can be rescued by wild-type Prpf31. Transgenic retina-specific expression of AD5 reveals that stable AD5 protein is initially localized in the nucleus but later found in the cytoplasm concurrent with progressing rod outer segment degeneration and apoptosis. Importantly, we show for the first time in vivo that retinal transcripts are wrongly spliced in adult transgenic retinas expressing AD5 and exhibiting increased apoptosis in rod photoreceptors. Conclusion: Our data suggest that distinct mutations in Prpf31 can lead to photoreceptor degeneration through different mechanisms, by haploinsufficiency or dominant-negative effects. Analyzing the AD5 effects in our animal model in vivo, our data imply that aberrant splicing of distinct retinal transcripts contributes to the observed retina defects.}, language = {en} } @article{YilmazRoeschKlingeletal.2011, author = {Yilmaz, Ali and R{\"o}sch, Sabine and Klingel, Karin and Kandolf, Reinhard and Helluy, Xavier and Hiller, Karl-Heinz and Jakob, Peter M and Sechtem, Udo}, title = {Molecular magnetic resonance imaging (MRI) of inflamed myocardium using ferucarbotran in patients with acute myocardial infarction}, series = {Journal of Cardiovascular Magnetic Resonance}, volume = {13}, journal = {Journal of Cardiovascular Magnetic Resonance}, number = {Suppl. 1}, doi = {10.1186/1532-429X-13-S1-P149}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-140991}, pages = {P149}, year = {2011}, abstract = {Introduction: Superparamagnetic iron oxide nanoparticle (SPIO)-based molecular imaging agents targeting macrophages have been developed and successfully applied in animal models of myocardial infarction.}, language = {en} } @article{WolterEndesfelderLindeetal.2011, author = {Wolter, Steve and Endesfelder, Ulrike and Linde, Sebastian van de and Heilemann, Mike and Sauer, Markus}, title = {Measuring localization performance of super-resolution algorithms on very active samples}, series = {Optics Express}, journal = {Optics Express}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-85936}, year = {2011}, abstract = {Super-resolution fluorescence imaging based on inglemolecule localization relies critically on the availability of efficient processing algorithms to distinguish, identify, and localize emissions of single fluorophores. In multiple current applications, such as threedimensional, time-resolved or cluster imaging, high densities of fluorophore emissions are common. Here, we provide an analytic tool to test the performance and quality of localization microscopy algorithms and demonstrate that common algorithms encounter difficulties for samples with high fluorophore density. We demonstrate that, for typical single-molecule localization microscopy methods such as dSTORM and the commonly used rapidSTORM scheme, computational precision limits the acceptable density of concurrently active fluorophores to 0.6 per square micrometer and that the number of successfully localized fluorophores per frame is limited to 0.2 per square micrometer.}, language = {en} } @phdthesis{Wolski2011, author = {Wolski, Stefanie Carola}, title = {Structural and functional characterization of nucleotide excision repair proteins}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-67183}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2011}, abstract = {XPD is a 5'-3' helicase of the superfamily 2. As part of the transcription factor IIH it functions in transcription initiation and nucleotide excision repair. This work focus on the role of XPD in nucleotide excision repair. NER is a DNA repair pathway unique for its broad substrate range. In placental mammals NER is the only repair mechanism able to remove lesions induced by UV-light. NER can be divided into four different steps that are conserved between pro- and eukaryotes. Step 1 consists of the initial damage recognition, during step 2 the putative damage is verified, in step 3 the verified damage is excised and in the 4th and final step the resulting gap in the DNA is refilled. XPD was shown to be involved in the damage verification step. It was possible to solve the first apo XPD structure by a MAD approach using only the endogenous iron from the iron sulfur cluster. Based on the apo XPD structure several questions arise: where is DNA bound? Where is DNA separated? How is damage verification achieved? What is the role of the FeS cluster? These questions were addressed in this work. Hypothesis driven structure based functional mutagenesis was employed and combined with detailed biochemical characterization of the variants. The variants were analyzed by thermal unfolding studies to exclude the possibility that the overall stability could be affected by the point mutation. DNA binding assays, ATPase assays and helicase assays were performed to delineate amino acid residues important for DNA binding, helicase activity and damage recognition. A structure of XPD containing a four base pair DNA fragment was solved by molecular replacement. This structure displays the polarity of the translocated strand with respect to the helicase framework. Moreover the properties of the FeS cluster were studied by electron paramagnetic resonance to get insights into the role of the FeS cluster. Furthermore XPD from Ferroplasma acidarmanus was investigated since it was shown that it is stalled at CPD containing lesions. The data provide the first detailed insight into the translocation mechanism of a SF2B helicase and reveal how polarity is achieved. This provides a basis for further anlayses understanding the combined action of the helicase and the 4Fe4S cluster to accomplish damage verification within the NER cascade.}, subject = {DNS-Reparatur}, language = {en} } @article{WobserGaiglTrautmann2011, author = {Wobser, Marion and Gaigl, Zeno and Trautmann, Axel}, title = {The concept of "compartment allergy": prilocaine injected into different skin layers}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-68679}, year = {2011}, abstract = {We herein present a patient with delayed-type allergic hypersensitivity against prilocaine leading to spreading eczematous dermatitis after subcutaneous injections for local anesthesia with prilocaine. Prilocaine allergy was proven by positive skin testing and subcutaneous provocation, whereas the evaluation of other local anesthetics - among them lidocaine, articaine and mepivacaine - did not exhibit any evidence for cross-reactivity. Interestingly, our patient repeatedly tolerated strictly deep subcutaneous injection of prilocaine in provocation testing while patch and superficial subcutaneous application mounted strong allergic responses. We hypothesize, that lower DC density in deeper cutaneous compartments and/or different DC subsets exhibiting distinct functional immunomodulatory properties in the various layers of the skin may confer to the observed absence of clinical reactivity against prilocaine after deep subcutaneous injection. The term compartment allergy indicates that the route of allergen administration together with the targeted immunologic environment orchestrates on the immunologic outcome: overt T-cell mediated allergy or clinical tolerance.}, subject = {Medizin}, language = {en} } @article{WippelFoertschHuppetal.2011, author = {Wippel, Carolin and F{\"o}rtsch, Christina and Hupp, Sabrina and Maier, Elke and Benz, Roland and Ma, Jiangtao and Mitchell, Timothy J and Iliev, Asparouh I}, title = {Extracellular Calcium Reduction Strongly Increases the Lytic Capacity of Pneumolysin From Streptococcus Pneumoniae in Brain Tissue}, series = {The Journal of Infectious Diseases}, volume = {204}, journal = {The Journal of Infectious Diseases}, number = {6}, doi = {10.1093/infdis/jir434}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-139356}, pages = {930-936}, year = {2011}, abstract = {Background Streptococcus pneumoniae causes serious diseases such as pneumonia and meningitis. Its major pathogenic factor is the cholesterol-dependent cytolysin pneumolysin, which produces lytic pores at high concentrations. At low concentrations, it has other effects, including induction of apoptosis. Many cellular effects of pneumolysin appear to be calcium dependent. Methods  Live imaging of primary mouse astroglia exposed to sublytic amounts of pneumolysin at various concentrations of extracellular calcium was used to measure changes in cellular permeability (as judged by lactate dehydrogenase release and propidium iodide chromatin staining). Individual pore properties were analyzed by conductance across artificial lipid bilayer. Tissue toxicity was studied in continuously oxygenated acute brain slices. Results  The reduction of extracellular calcium increased the lytic capacity of the toxin due to increased membrane binding. Reduction of calcium did not influence the conductance properties of individual toxin pores. In acute cortical brain slices, the reduction of extracellular calcium from 2 to 1 mM conferred lytic activity to pathophysiologically relevant nonlytic concentrations of pneumolysin. Conclusions  Reduction of extracellular calcium strongly enhanced the lytic capacity of pneumolysin due to increased membrane binding. Thus, extracellular calcium concentration should be considered as a factor of primary importance for the course of pneumococcal meningitis. "}, language = {en} } @article{WilliamsMachannKuehleretal.2011, author = {Williams, Tatjana and Machann, Wolfram and K{\"u}hler, Leif and Hamm, Henning and M{\"u}ller-H{\"o}cker, Josef and Zimmer, Michael and Ertl, Georg and Ritter, Oliver and Beer, Meinrad and Sch{\"o}nberger, Jost}, title = {Novel desmoplakin mutation: juvenile biventricular cardiomyopathy with left ventricular non-compaction and acantholytic palmoplantar keratoderma}, series = {Clinical Research in Cardiology}, volume = {100}, journal = {Clinical Research in Cardiology}, number = {12}, doi = {10.1007/s00392-011-0345-9}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-141198}, pages = {1087-1093}, year = {2011}, abstract = {Two sons of a consanguineous marriage developed biventricular cardiomyopathy. One boy died of severe heart failure at the age of 6 years, the other was transplanted because of severe heart failure at the age of 10 years. In addition, focal palmoplantar keratoderma and woolly hair were apparent in both boys. As similar phenotypes have been described in Naxos disease and Carvajal syndrome, respectively, the genes for plakoglobin (JUP) and desmoplakin (DSP) were screened for mutations using direct genomic sequencing. A novel homozygous 2 bp deletion was identified in an alternatively spliced region of DSP. The deletion 5208_5209delAG led to a frameshift downstream of amino acid 1,736 with a premature truncation of the predominant cardiac isoform DSP-1. This novel homozygous truncating mutation in the isoform-1 specific region of the DSP C-terminus caused Carvajal syndrome comprising severe early-onset heart failure with features of non-compaction cardiomyopathy, woolly hair and an acantholytic form of palmoplantar keratoderma in our patient. Congenital hair abnormality and manifestation of the cutaneous phenotype in toddler age can help to identify children at risk for cardiac death.}, language = {en} } @article{WiegeringSchickBeeretal.2011, author = {Wiegering, Verena and Schick, Judith and Beer, Meinrad and Gattenl{\"o}hner, Stefan and Girschick, Hermann and Liese, Johannes and Schlegel, Paul and Eyrich, Matthias}, title = {Varicella-zoster virus infections in immunocompromised patients - a single centre 6-years analysis}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-68723}, year = {2011}, abstract = {Background: Infection with varicella-zoster virus (VZV) contemporaneously with malignant disease or immunosuppression represents a particular challenge and requires individualized decisions and treatment. Although the increasing use of varicella-vaccines in the general population and rapid initiation of VZVimmunoglobulins and acyclovir in case of exposure has been beneficial for some patients, immunocompromised individuals are still at risk for unfavourable courses. Methods: In this single center, 6-year analysis we review incidence, hospitalization and complication rates of VZVinfections in our center and compare them to published data. Furthermore, we report three instructive cases. Results: Hospitalization rate of referred children with VZV-infections was 45\%, among these 17\% with malignancies and 9\% under immunosuppressive therapy. Rate of complications was not elevated in these two high-risk cohorts, but one ALL-patient died due to VZV-related complications. We report one 4-year old boy with initial diagnosis of acute lymphoblastic leukemia who showed a rapidly fatal outcome of his simultaneous varicella-infection, one 1.8-year old boy with an identical situation but a mild course of his disease, and an 8.5-year old boy with a steroiddependent nephrotic syndrome. This boy developed severe hepatic involvement during his varicella-infection but responded to immediate withdrawl of steroids and administration of acyclovir plus single-dose cidofovir after nonresponse to acyclovir after 48 h. Conclusion: Our data show that patients with malignant diseases or immunosuppressive therapy should be hospitalized and treated immediately with antiviral agents. Despite these measures the course of VZV-infections can be highly variable in these patients. We discuss aids to individual decision-making for these difficult situations.}, subject = {Varizellen-Virus}, language = {en} } @article{WernerSchwedeGenieseretal.2011, author = {Werner, Katharina and Schwede, Frank and Genieser, Hans-Gottfried and Geiger, J{\"o}rg and Butt, Elke}, title = {Quantification of cAMP and cGMP analogs in intact cells: pitfalls in enzyme immunoassays for cyclic nucleotides}, series = {Naunyn-Schmiedeberg's Archives of Pharmacology}, volume = {384}, journal = {Naunyn-Schmiedeberg's Archives of Pharmacology}, number = {2}, doi = {10.1007/s00210-011-0662-6}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-141828}, pages = {169-176}, year = {2011}, abstract = {Immunoassays are routinely used as research tools to measure intracellular cAMP and cGMP concentrations. Ideally, this application requires antibodies with high sensitivity and specificity. The present work evaluates the cross-reactivity of commercially available cyclic nucleotide analogs with two non-radioactive and one radioactive cAMP and cGMP immunoassay. Most of the tested cyclic nucleotide analogs showed low degree competition with the antibodies; however, with Rp-cAMPS, 8-Br-cGMP and 8-pCPT-cGMP, a strong cross-reactivity with the corresponding cAMP and cGMP, respectively, immunoassays was observed. The determined EIA-binding constants enabled the measurement of the intracellular cyclic nucleotide concentrations and revealed a time- and lipophilicity-dependent cell membrane permeability of the compounds in the range of 10-30\% of the extracellular applied concentration, thus allowing a more accurate prediction of the intracellular analog levels in a given experiment.}, language = {en} } @misc{Wenzel2011, type = {Master Thesis}, author = {Wenzel, Frank}, title = {Smell and repel: Resin based defense mechanisms and interactions between Australian ants and stingless bees}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-65960}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2011}, abstract = {Bees are subject to permanent threat from predators such as ants. Their nests with large quantities of brood, pollen and honey represent lucrative targets for attacks whereas foragers have to face rivalry at food sources. This thesis focused on the role of stingless bees as third party interactor on ant-aphid-associations as well as on the predatory potential represented by ants and defense mechanisms against this threat. Regular observations of an aphid infested Podocarpus for approaching stingless bees yielded no results. Another aim of this thesis was the observation of foraging habits of four native and one introduced ant species for assessment of their predatory potential to stingless bees. All species turned out to be dietary balanced generalists with one mostly carnivorous species and four species predominantly collecting nectar roughly according to optimal foraging theory. Two of the species monitored, Rhytidoponera metallica and Iridomyrmex rufoniger were considered potential nest robbers. As the name implies, stingless bees lack the powerful weapon of their distant relatives; hence they specialized on other defense strategies. Resin is an important, multipurpose resource for stingless bees that is used as material for nest construction, antibiotic and for defensive means. For the latter purpose highly viscous resin is either directly used to stick down aggressors or its terpenic compounds are included in the bees cuticular surface. In a feeding choice experiment, three ant species were confronted with the choice between two native bee species - Tetragonula carbonaria and Austroplebeia australis - with different cuticular profiles and resin collection habits. Two of the ant species, especially the introduced Tetramorium bicarinatum did not show any preferences. The carnivorous R. metallica predominantly took the less resinous A. australis as prey. The reluctance towards T. carbonaria disappeared when the resinous compounds on its cuticle had been washed off with hexane. To test whether the repulsive reactions were related to the stickiness of the resinous surface or to chemical substances, hexane extracts of bees' cuticles, propolis and three natural tree resins were prepared. In the following assay responses of ants towards extract treated surfaces were observed. Except for one of the resin extracts, all tested substances had repellent effects to the ants. Efficacy varied with the type of extract and species. Especially to the introduced T. bicarinatum the cuticular extract had no effect. GCMS-analyses showed that some of the resinous compounds were also found in the cuticular profile of T. carbonaria which featured reasonable analogies to the resin of Corymbia torelliana that is highly attractive for stingless bees. The results showed that repellent effects were only partially related to the sticky quality of resin but were rather caused by chemical substances, presumably sesqui- and diterpenes. Despite its efficacy this defense strategy only provides short time repellent effects sufficient for escape and warning of nest mates to initiate further preventive measures.}, subject = {Stachellose Biene}, language = {en} } @phdthesis{Weissflog2011, author = {Weißflog, Lena}, title = {Molecular Genetics of Emotional Dysregulation in Attention-Deficit/Hyperactivity Disorder}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-69345}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2011}, abstract = {Attention-deficit/hyperactivity disorder (ADHD) is a genetically complex childhood onset neurodevelopmental disorder which is highly persistent into adulthood. Several chromo-somal regions associated with this disorder were identified previously in genome-wide linkage scans, association (GWA) and copy number variation (CNV) studies. In this work the results of case-control and family-based association studies using a can-didate gene approach are presented. For this purpose, possible candidate genes for ADHD have been finemapped using mass array-based SNP genotyping. The genes KCNIP4, CDH13 and DIRAS2 have been found to be associated with ADHD and, in addition, with cluster B and cluster C personality disorders (PD) which are known to be related to ADHD. Most of the associations found in this work would not withstand correction for multiple testing. However, a replication in several independent populations has been achieved and in conjunction with previous evidence from linkage, GWA and CNV studies, it is assumed that there are true associations between those genes and ADHD. Further investigation of DIRAS2 by quantitative real-time PCR (qPCR) revealed expression in the hippocampus, cerebral cortex and cerebellum of the human brain and a significant increase in Diras2 expression in the mouse brain during early development. In situ hybrid-izations on murine brain slices confirmed the results gained by qPCR in the human brain. Moreover, Diras2 is expressed in the basolateral amygdala, structures of the olfactory system and several other brain regions which have been implicated in the psychopatholo-gy of ADHD. In conclusion, the results of this work provide further support to the existence of a strong genetic component in the pathophysiology of ADHD and related disorders. KCNIP4, CDH13 and DIRAS2 are promising candidates and need to be further examined to get more knowledge about the neurobiological basis of this common disease. This knowledge is essential for understanding the molecular mechanisms underlying the emergence of this disorder and for the development of new treatment strategies.}, subject = {Aufmerksamkeits-Defizit-Syndrom}, language = {en} } @phdthesis{Weiss2011, author = {Weiß, Sabine}, title = {Function of the Spir actin nucleators in intracellular vesicle transport processes}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-64589}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2011}, abstract = {Spir proteins are the founding members of the novel class of WH2-actin nucleators. A C-terminal modified FYVE zinc finger motif is necessary to target Spir proteins towards intracellular membranes. The function and regulation of the Spir actin organizers at vesicular membranes is almost unknown. Live cell imaging analyses performed in this study show that Spir-2 is localized at tubular vesicles. Cytoplasmic Spir-2-associated vesicles branch and form protrusions, which can make contacts to the microtubule network, where the Spir-2 vesicles stretch and slide along the microtubule filaments. The analysis of living HeLa cells expressing eGFP-tagged Spir-2, Spir-2-ΔKIND and Spir-2-ΔKW (lacking the 4 WH2 domains and the KIND domain) showed Spir-2-associated tubular structures which differ in their length and motility. Throughout the course of that study it could be shown that the tail domain of the actin motor protein myosin Vb, as a force-generating molecule, is colocalizing and co-immunoprecipitating with Spir-2-ΔKW. By using the tail domain of myosin Vb as a dominant negative mutant for myosin Vb-dependent vesicle transport processes it could be shown that Spir-2-ΔKW/MyoVb-cc-tail- associated vesicles exhibit an increased elongation. Moreover, using the microtubule depolymerizing drug nocodazole it could be shown that the elongation and the motility of Spir-2-ΔKW-associated vesicles depends on an intact microtubule cytoskeleton. Motility and morphological dynamics of Spir-2-associated vesicles is therefore dependent on actin, actin motorproteins and microtubule filaments. These results propose a model in which myosin/F-actin forces mediate vesicle branching, allowing the vesicles to move to and in between the microtubule filaments and thereby providing a new degree of freedom in vesicular motility. To determine the exact subcellular localization of Spir-2, colocalization studies were performed. It could be shown that Spir-2 shows a partial colocalization to Rab11a-positive compartments. Furthermore, Spir-2 exhibits an almost identical localization to Arf1 and the Arf1 small G protein but not Rab11a could be immunoprecipitated with Spir-2-ΔKW. This suggests, that Arf1 recruits Spir-2 to Arf1/Rab11a-positive membranes. Another important function of the Spir-2 C-terminus is the membrane targeting by the FYVE domain. By performing a protein-lipid overlay assay, it has been shown that purified GST- and 6xHis-tagged Spir-2-ΔKW bind phosphatidic acid suggesting a mechanism in which Spir-2 is recruited to phosphatidic acid-enriched membranes. To further elucidate the mechanism in which Spir-2 membrane-targeting could be regulated, interaction studies of C-terminal parts of Spir-2 revealed that the Spir-2 proteins interact directly.}, subject = {Aktin}, language = {en} } @article{WeiseBasseLuesebrinkKleinschnitzetal.2011, author = {Weise, Gesa and Basse-L{\"u}sebrink, Thomas C. and Kleinschnitz, Christoph and Kampf, Thomas and Jakob, Peter M. and Stoll, Guido}, title = {In Vivo Imaging of Stepwise Vessel Occlusion in Cerebral Photothrombosis of Mice by \(^{19}\)F MRI}, series = {PLoS One}, volume = {6}, journal = {PLoS One}, number = {12}, doi = {10.1371/journal.pone.0028143}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-137792}, pages = {e28143}, year = {2011}, abstract = {Background \(^{19}\)F magnetic resonance imaging (MRI) was recently introduced as a promising technique for in vivo cell tracking. In the present study we compared \(^{19}\)F MRI with iron-enhanced MRI in mice with photothrombosis (PT) at 7 Tesla. PT represents a model of focal cerebral ischemia exhibiting acute vessel occlusion and delayed neuroinflammation. Methods/Principal Findings Perfluorocarbons (PFC) or superparamagnetic iron oxide particles (SPIO) were injected intravenously at different time points after photothrombotic infarction. While administration of PFC directly after PT induction led to a strong \(^{19}\)F signal throughout the entire lesion, two hours delayed application resulted in a rim-like \(^{19}\)F signal at the outer edge of the lesion. These findings closely resembled the distribution of signal loss on T2-weighted MRI seen after SPIO injection reflecting intravascular accumulation of iron particles trapped in vessel thrombi as confirmed histologically. By sequential administration of two chemically shifted PFC compounds 0 and 2 hours after illumination the different spatial distribution of the \(^{19}\)F markers (infarct core/rim) could be visualized in the same animal. When PFC were applied at day 6 the fluorine marker was only detected after long acquisition times ex vivo. SPIO-enhanced MRI showed slight signal loss in vivo which was much more prominent ex vivo indicative for neuroinflammation at this late lesion stage. Conclusion Our study shows that vessel occlusion can be followed in vivo by \(^{19}\)F and SPIO-enhanced high-field MRI while in vivo imaging of neuroinflammation remains challenging. The timing of contrast agent application was the major determinant of the underlying processes depicted by both imaging techniques. Importantly, sequential application of different PFC compounds allowed depiction of ongoing vessel occlusion from the core to the margin of the ischemic lesions in a single MRI measurement.}, language = {en} } @article{WeisSchoenVictoretal.2011, author = {Weis, Eva and Schoen, Holger and Victor, Anja and Spix, Claudia and Ludwig, Marco and Schneider-Raetzke, Brigitte and Kohlschmidt, Nicolai and Bartsch, Oliver and Gerhold-Ay, Aslihan and Boehm, Nils and Grus, Franz and Haaf, Thomas and Galetzka, Danuta}, title = {Reduced mRNA and Protein Expression of the Genomic Caretaker RAD9A in Primary Fibroblasts of Individuals with Childhood and Independent Second Cancer}, series = {PLoS ONE}, volume = {6}, journal = {PLoS ONE}, number = {10}, doi = {10.1371/journal.pone.0025750}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-141838}, pages = {e25750}, year = {2011}, abstract = {Background: The etiology of secondary cancer in childhood cancer survivors is largely unclear. Exposure of normal somatic cells to radiation and/or chemotherapy can damage DNA and if not all DNA lesions are properly fixed, the mis-repair may lead to pathological consequences. It is plausible to assume that genetic differences, i.e. in the pathways responsible for cell cycle control and DNA repair, play a critical role in the development of secondary cancer. Methodology/Findings: To identify factors that may influence the susceptibility for second cancer formation, we recruited 20 individuals who survived a childhood malignancy and then developed a second cancer as well as 20 carefully matched control individuals with childhood malignancy but without a second cancer. By antibody microarrays, we screened primary fibroblasts of matched patients for differences in the amount of representative DNA repair-associated proteins. We found constitutively decreased levels of RAD9A and several other DNA repair proteins in two-cancer patients, compared to one-cancer patients. The RAD9A protein level increased in response to DNA damage, however to a lesser extent in the two-cancer patients. Quantification of mRNA expression by real-time RT PCR revealed lower RAD9A mRNA levels in both untreated and 1 Gy gamma-irradiated cells of two-cancer patients. Conclusions/Significance: Collectively, our results support the idea that modulation of RAD9A and other cell cycle arrest and DNA repair proteins contribute to the risk of developing a second malignancy in childhood cancer patients.}, language = {en} } @article{WeisSchoenVictoretal.2011, author = {Weis, Eva and Schoen, Holger and Victor, Anja and Spix, Claudia and Ludwig, Marco and Schneider-Raetzke, Brigitte and Kohlschmidt, Nicolai and Bartsch, Oliver and Gerhold-Ay, Aslihan and Boehm, Nils and Grus, Franz and Haaf, Thomas and Galetzka, Danuta}, title = {Reduced mRNA and Protein Expression of the Genomic Caretaker RAD9A in Primary Fibroblasts of Individuals with Childhood and Independent Second Cancer}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-74777}, year = {2011}, abstract = {Background: The etiology of secondary cancer in childhood cancer survivors is largely unclear. Exposure of normal somatic cells to radiation and/or chemotherapy can damage DNA and if not all DNA lesions are properly fixed, the mis-repair may lead to pathological consequences. It is plausible to assume that genetic differences, i.e. in the pathways responsible for cell cycle control and DNA repair, play a critical role in the development of secondary cancer. Methodology/Findings: To identify factors that may influence the susceptibility for second cancer formation, we recruited 20 individuals who survived a childhood malignancy and then developed a second cancer as well as 20 carefully matched control individuals with childhood malignancy but without a second cancer. By antibody microarrays, we screened primary fibroblasts of matched patients for differences in the amount of representative DNA repair-associated proteins. We found constitutively decreased levels of RAD9A and several other DNA repair proteins in two-cancer patients, compared to onecancer patients. The RAD9A protein level increased in response to DNA damage, however to a lesser extent in the twocancer patients. Quantification of mRNA expression by real-time RT PCR revealed lower RAD9A mRNA levels in both untreated and 1 Gy c-irradiated cells of two-cancer patients. Conclusions/Significance: Collectively, our results support the idea that modulation of RAD9A and other cell cycle arrest and DNA repair proteins contribute to the risk of developing a second malignancy in childhood cancer patients.}, subject = {Medizin}, language = {en} } @article{WeibelRaabYuetal.2011, author = {Weibel, Stephanie and Raab, Viktoria and Yu, Yong A. and Worschech, Andrea and Wang, Ena and Marincola, Francesco M. and Szalay, Aladar A.}, title = {Viral-mediated oncolysis is the most critical factor in the late-phase of the tumor regression process upon vaccinia virus infection}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-68691}, year = {2011}, abstract = {Background: In principle, the elimination of malignancies by oncolytic virotherapy could proceed by different mechanisms - e.g. tumor cell specific oncolysis, destruction of the tumor vasculature or an anti-tumoral immunological response. In this study, we analyzed the contribution of these factors to elucidate the responsible mechanism for regression of human breast tumor xenografts upon colonization with an attenuated vaccinia virus (VACV). Methods: Breast tumor xenografts were analyzed 6 weeks post VACV infection (p.i.; regression phase) by immunohistochemistry and mouse-specific expression arrays. Viral-mediated oncolysis was determined by tumor growth analysis combined with microscopic studies of intratumoral virus distribution. The tumor vasculature was morphologically characterized by diameter and density measurements and vessel functionality was analyzed by lectin perfusion and extravasation studies. Immunological aspects of viral-mediated tumor regression were studied in either immune-deficient mouse strains (T-, B-, NK-cell-deficient) or upon cyclophosphamide-induced immunosuppression (MHCII+-cell depletion) in nude mice. Results: Late stage VACV-infected breast tumors showed extensive necrosis, which was highly specific to cancer cells. The tumor vasculature in infected tumor areas remained functional and the endothelial cells were not infected. However, viral colonization triggers hyperpermeability and dilatation of the tumor vessels, which resembled the activated endothelium in wounded tissue. Moreover, we demonstrated an increased expression of genes involved in leukocyte-endothelial cell interaction in VACV-infected tumors, which orchestrate perivascular inflammatory cell infiltration. The immunohistochemical analysis of infected tumors displayed intense infiltration of MHCII-positive cells and colocalization of tumor vessels with MHCII+/CD31+ vascular leukocytes. However, GI-101A tumor growth analysis upon VACV-infection in either immunosuppressed nude mice (MHCII+-cell depleted) or in immune-deficient mouse strains (T-, B-, NK-cell-deficient) revealed that neither MHCII-positive immune cells nor T-, B-, or NK cells contributed significantly to VACV-mediated tumor regression. In contrast, tumors of immunosuppressed mice showed enhanced viral spreading and tumor necrosis. Conclusions: Taken together, these results indicate that VACV-mediated oncolysis is the primary mechanism of tumor shrinkage in the late regression phase. Neither the destruction of the tumor vasculature nor the massive VACV-mediated intratumoral inflammation was a prerequisite for tumor regression. We propose that approaches to enhance viral replication and spread within the tumor microenvironment should improve therapeutical outcome.}, subject = {Virusinfektion}, language = {en} } @article{WehnerWeisteDroegeLaser2011, author = {Wehner, Nora and Weiste, Christoph and Dr{\"o}ge-Laser, Wolfgang}, title = {Molecular screening tools to study Arabidopsis transcription factors}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-69226}, year = {2011}, abstract = {In the model plant Arabidopsis thaliana, more than 2000 genes are estimated to encode transcription factors (TFs), which clearly emphasizes the importance of transcriptional control. Although genomic approaches have generated large TF open reading frame (ORF) collections, only a limited number of these genes is functionally characterized, yet. This review evaluates strategies and methods to identify TF functions. In particular, we focus on two recently developed TF screening platforms, which make use of publically available GATEWAY®-compatible ORF collections. (1) The Arabidopsis thaliana TF ORF over-Expression (AtTORF-Ex) library provides pooled collections of transgenic lines over-expressing HA-tagged TF genes, which are suited for screening approaches to define TF functions in stress defense and development. (2) A high-throughput microtiter plate based protoplast trans activation (PTA) system has been established to screen for TFs which are regulating a given promoter:Luciferase construct in planta.}, subject = {Biologie}, language = {en} } @article{WegertBausenweinKneitzetal.2011, author = {Wegert, Jenny and Bausenwein, Sabrina and Kneitz, Susanne and Roth, Sabine and Graf, Norbert and Geissinger, Eva and Gessler, Manfred}, title = {Retinoic acid pathway activity in Wilms tumors and characterization of biological responses in vitro}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-69137}, year = {2011}, abstract = {Background: Wilms tumor (WT) is one of the most common malignancies in childhood. With current therapy protocols up to 90\% of patients can be cured, but there is still a need to improve therapy for patients with aggressive WT and to reduce treatment intensity where possible. Prior data suggested a deregulation of the retinoic acid (RA) signaling pathway in high-risk WT, but its mode of action remained unclear. Results: The association of retinoid signaling and clinical parameters could be validated in a large independent tumor set, but its relevance in primary nephrectomy tumors from very young children may be different. Reduced RA pathway activity and MYCN overexpression were found in high risk tumors as opposed to tumors with low/ intermediate risk, suggesting a beneficial impact of RA especially on advanced WT. To search for possible modes of action of retinoids as novel therapeutic options, primary tumor cell cultures were treated in vitro with all-trans-RA (ATRA), 9cis-RA, fenretinide and combinations of retinoids and a histone deacetylase (HDAC) inhibitor. Genes deregulated in high risk tumors showed opposite changes upon treatment suggesting a positive effect of retinoids. 6/7 primary cultures tested reduced proliferation, irrespective of prior RA signaling levels. The only variant culture was derived from mesoblastic nephroma, a distinct childhood kidney neoplasm. Retinoid/HDAC inhibitor combinations provided no synergistic effect. ATRA and 9cis-RA induced morphological changes suggestive of differentiation, while fenretinide induced apoptosis in several cultures tested. Microarray analysis of ATRA treated WT cells revealed differential expression of many genes involved in extracellular matrix formation and osteogenic, neuronal or muscle differentiation. The effects documented appear to be reversible upon drug withdrawal, however. Conclusions: Altered retinoic acid signaling has been validated especially in high risk Wilms tumors. In vitro testing of primary tumor cultures provided clear evidence of a potential utility of retinoids in Wilms tumor treatment based on the analysis of gene expression, proliferation, differentiation and apoptosis.}, subject = {Krebs}, language = {en} } @article{WedelHudakSeibeletal.2011, author = {Wedel, Steffen and Hudak, Lukasz and Seibel, Jens-Michael and Makarevic, Jasmina and Juengel, Eva and Tsaur, Igor and Waaga-Gasser, Ana and Haferkamp, Axel and Blaheta, Roman A.}, title = {Molecular targeting of prostate cancer cells by a triple drug combination down-regulates integrin driven adhesion processes, delays cell cycle progression and interferes with the cdk-cyclin axis}, series = {BMC Cancer}, volume = {11}, journal = {BMC Cancer}, number = {375}, doi = {10.1186/1471-2407-11-375}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-141075}, pages = {1-14}, year = {2011}, abstract = {Background: Single drug use has not achieved satisfactory results in the treatment of prostate cancer, despite application of increasingly widespread targeted therapeutics. In the present study, the combined impact of the mammalian target of rapamycin (mTOR)-inhibitor RAD001, the dual EGFr and VGEFr tyrosine kinase inhibitor AEE788 and the histone deacetylase (HDAC)-inhibitor valproic acid (VPA) on prostate cancer growth and adhesion in vitro was investigated. Methods: PC-3, DU-145 and LNCaP cells were treated with RAD001, AEE788 or VPA or with a RAD-AEE-VPA combination. Tumor cell growth, cell cycle progression and cell cycle regulating proteins were then investigated by MTT-assay, flow cytometry and western blotting, respectively. Furthermore, tumor cell adhesion to vascular endothelium or to immobilized extracellular matrix proteins as well as migratory properties of the cells was evaluated, and integrin alpha and beta subtypes were analyzed. Finally, effects of drug treatment on cell signaling pathways were determined. Results: All drugs, separately applied, reduced tumor cell adhesion, migration and growth. A much stronger anticancer effect was evoked by the triple drug combination. Particularly, cdk1, 2 and 4 and cyclin B were reduced, whereas p27 was elevated. In addition, simultaneous application of RAD001, AEE788 and VPA altered the membranous, cytoplasmic and gene expression pattern of various integrin alpha and beta subtypes, reduced integrin-linked kinase (ILK) and deactivated focal adhesion kinase (FAK). Signaling analysis revealed that EGFr and the downstream target Akt, as well as p70S6k was distinctly modified in the presence of the drug combination. Conclusions: Simultaneous targeting of several key proteins in prostate cancer cells provides an advantage over targeting a single pathway. Since strong anti-tumor properties became evident with respect to cell growth and adhesion dynamics, the triple drug combination might provide progress in the treatment of advanced prostate cancer.}, language = {en} } @article{WangorschButtMarketal.2011, author = {Wangorsch, Gaby and Butt, Elke and Mark, Regina and Hubertus, Katharina and Geiger, J{\"o}rg and Dandekar, Thomas and Dittrich, Marcus}, title = {Time-resolved in silico modeling of fine-tuned cAMP signaling in platelets: feedback loops, titrated phosphorylations and pharmacological modulation}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-69145}, year = {2011}, abstract = {Background: Hemostasis is a critical and active function of the blood mediated by platelets. Therefore, the prevention of pathological platelet aggregation is of great importance as well as of pharmaceutical and medical interest. Endogenous platelet inhibition is predominantly based on cyclic nucleotides (cAMP, cGMP) elevation and subsequent cyclic nucleotide-dependent protein kinase (PKA, PKG) activation. In turn, platelet phosphodiesterases (PDEs) and protein phosphatases counterbalance their activity. This main inhibitory pathway in human platelets is crucial for countervailing unwanted platelet activation. Consequently, the regulators of cyclic nucleotide signaling are of particular interest to pharmacology and therapeutics of atherothrombosis. Modeling of pharmacodynamics allows understanding this intricate signaling and supports the precise description of these pivotal targets for pharmacological modulation. Results: We modeled dynamically concentration-dependent responses of pathway effectors (inhibitors, activators, drug combinations) to cyclic nucleotide signaling as well as to downstream signaling events and verified resulting model predictions by experimental data. Experiments with various cAMP affecting compounds including antiplatelet drugs and their combinations revealed a high fidelity, fine-tuned cAMP signaling in platelets without crosstalk to the cGMP pathway. The model and the data provide evidence for two independent feedback loops: PKA, which is activated by elevated cAMP levels in the platelet, subsequently inhibits adenylyl cyclase (AC) but as well activates PDE3. By multi-experiment fitting, we established a comprehensive dynamic model with one predictive, optimized and validated set of parameters. Different pharmacological conditions (inhibition, activation, drug combinations, permanent and transient perturbations) are successfully tested and simulated, including statistical validation and sensitivity analysis. Downstream cyclic nucleotide signaling events target different phosphorylation sites for cAMP- and cGMP-dependent protein kinases (PKA, PKG) in the vasodilator-stimulated phosphoprotein (VASP). VASP phosphorylation as well as cAMP levels resulting from different drug strengths and combined stimulants were quantitatively modeled. These predictions were again experimentally validated. High sensitivity of the signaling pathway at low concentrations is involved in a fine-tuned balance as well as stable activation of this inhibitory cyclic nucleotide pathway. Conclusions: On the basis of experimental data, literature mining and database screening we established a dynamic in silico model of cyclic nucleotide signaling and probed its signaling sensitivity. Thoroughly validated, it successfully predicts drug combination effects on platelet function, including synergism, antagonism and regulatory loops.}, subject = {Vasodilatator-stimuliertes Phosphoprotein}, language = {en} } @phdthesis{Wang2011, author = {Wang, Huiqiang}, title = {Enhanced Replication of Vaccinia Virus GLV-1h68 in Cancer Stem-like Cells of Human Breast Cancer Cell Preparations}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-64750}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2011}, abstract = {There is more and more evidence for the cancer stem cell hypothesis which believes that cancers are driven by a cellular subcomponent that has stem cell properties which is self-renewal, tumorigenicity and multilineage differentiation capacity. Cancer stem cells have been connected to the initiation of tumors and are even found to be responsible for relapses after apparently curative therapies have been undertaken. This hypothesis changes our conceptual approach of oncogenesis and shall have implications in breast cancer prevention, detection and treatment, especially in metastatic breast cancer for which no curative treatment exists. Given the specific stem cell features, novel therapeutic pathways can be targeted. Since the value of vaccinia virus as a vaccination virus against smallpox was discovered by E. Jenner at 18th century, it plays an important role in human medicine and molecular biology. After smallpox was successfully eradicated, vaccinia virus is mainly used as a viral vector in molecular biology and increasingly in cancer therapy. The outstanding capability to specifically target and destroy cancer cells makes it a perfect agent for oncolytic virotherapy. Furthermore, the virus can easily be modified by inserting genes which encode therapeutic or diagnostic proteins to be expressed when a tumor is infected. The emphasis in this study was the establishment of methods for the enrichment of human breast cancer stem-like cells from cancer cell lines and characterization of those cancer stem-like cells in vitro and in vivo. Furthermore, by using the Genelux Corporation vaccinia virus strain GLV-1h68, the isolated cancer stem-like cells can be targeted not only in vitro but also in vivo more efficiently. Side-population (SP) cells within cancers and cell lines are rare cell populations known to be enriched cancer stem-like cells. In this study, we used Hoechst 33342 staining and flow cytometry to identify SP cells from the human breast cancer cell lines MCF-7 and GI-101A as models for cancer stem-like cells. Considering the cytotoxicity of Hoechst dye and the restriction of instrument, we did not carry out further studies by this method. Utilizing in vitro and in vivo experimental systems, we showed that human breast cancer cell line GI-101A with aldehyde dehydrogenase activity (ALDH) have stemlike properties. Higher ALDH activity identifies the tumorigenic cell fraction which is capable of self-renewal and of generating tumors that could recapitulate the heterogeneity of the parental tumor. Furthermore, the cells with higher ALDH activity display significant resistance to chemotherapy and ionizing radiation, which proves their stem-like properties again. The cells which have higher ALDH activity also are more invasive compared to cells which have lower ALDH activity, which connects the cancer stem-like cells with cancer metastases. By analyzing the popular human breast cancer stem cells surface markers CD44, CD49f and CD24, it was discovered that the cells with higher ALDH activity have stronger CD44 and CD49f expression than in those cells with lower ALDH activity, which further confirms their stem-like properties. Finally, the cells with higher ALDH activity and lower ALDH activity were infected in vitro and used in virotherapy in a mouse xenograft model was performed. The results indicated that the vaccinia virus GLV-1h68 can replicate in cells with higher ALDH activity more efficiently than cells with lower ALDH activity. GLV-1h68 also can selectively target and eradicate the xenograft tumors which were derived from cells with higher ALDH activity. The epithelial-mesenchymal transition (EMT) is a key developmental program that is often activated during cancer invasion and metastases. EMT was induced in immortalized human mammary epithelial cells (HMLEs) and in GI-101A cells, which results in the acquisition of mesenchymal traits and in the expression of stem cell markers. Furthermore, the EMT-induced GI-101A cells showed resistance to chemotherapy and invasion capacity. CD44+/CD24- cells were enriched during the EMT induction. Following flow cytometry sorting by using CD44, CD24 and ESA surface marker, the sorted cells were tested in a mouse model regarding tumorigenicity. Unexpectedly, we found that CD44+/CD24+/ESA+ cells could initiate tumors more efficiently rather than CD44+/CD24-/ESA+ and other fractions in EMTinduced GI-101A cells. We also infected the CD44+/CD24+/ESA+ and CD44+/CD24- /ESA+ cells in vitro and performed virotherapy in a mouse xenograft model. The results indicated that the vaccinia virus GLV-1h68 is able to replicate in CD44+/CD24+/ESA+ cells more efficiently than in CD44+/CD24-/ESA+ cells. GLV-1h68 was also capable to selectively target and eradicate the xenograft tumors which derived from CD44+/CD24+/ESA+ cells. Moreover, CD44- cells have much lower tumorigenicity in the mouse model and CD44- cells derived-tumors are not responsive to vaccinia virotherapy. In summary, we have successfully established an in vitro and in vivo system for the identification, characterization and isolation of cancer stem-like cells from the human breast cancer cell line GI-101A by using the ALDEFLUOR assay. The vaccinia virus GLV-1h68 was able to efficiently target and eradicate the higher ALDH activity cells and tumors derived from those cells. Although contrary to the current assumption, CD44+/CD24+/ESA+ cells in the EMT-induced GI-101A cell line showed stem-like properties and GLV-1h68 was able to efficiently target and eradicate the CD44+/CD24+/ESA+ cells and tumors which derived from those cells. Finally, improved understanding of cancer stem cells may have tremendous relevance for how cancer should be treated. It is menacing that cancer stem cells are resistant to almost all anti-tumor approaches which have already been established for the treatment of metastatic diseases such as ionizing radiation, hormonal therapy, chemotherapy, and small molecular inhibitors. Therefore, it is promising that our results suggest that these cancer stem cells may be susceptible to treatment with oncolytic vaccinia virus.}, subject = {Vaccinia Virus}, language = {en} } @article{WalitzaMelfsenJansetal.2011, author = {Walitza, Susanne and Melfsen, Siebke and Jans, Thomas and Zellmann, Henrike and Wewetzer, Christoph and Warnke, Andreas}, title = {Obsessive-Compulsive Disorder in Children and Adolescents}, series = {Deutsches {\"A}rzteblatt International}, volume = {108}, journal = {Deutsches {\"A}rzteblatt International}, number = {11}, doi = {10.3238/arztebl.2011.0173}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-141214}, pages = {173-I}, year = {2011}, abstract = {Background: Early-onset obsessive-compulsive disorder (OCD) is one of the more common mental illnesses of children and adolescents, with prevalence of 1\% to 3\%. Its manifestations often lead to severe impairment and to conflict in the family. In this review, we summarize the manifestations, comorbidity, pathophysiology, and course of this disease as well as current modes of diagnosis and treatment. Methods: We selectively review the relevant literature and the German-language guidelines for the diagnosis and treatment of mental illnesses in children and adolescents. Results: Obsessive-compulsive manifestations are of many types and cause severe impairment. Comorbid mental disturbances are present in as many as 70\% of patients. The disease takes a chronic course in more than 40\% of patients. Cognitive behavioral therapy is the treatment of first choice, followed by combination pharmacotherapy including selective serotonin reuptake inhibitors (SSRI) and then by SSRI alone. Conclusion: OCD often begins in childhood or adolescence. There are empirically based neurobiological and cognitive-behavioral models of its pathophysiology. Multiaxial diagnostic evaluation permits early diagnosis. Behavioral therapy and medications are highly effective treatments, but the disorder nonetheless takes a chronic course in a large percentage of patients.}, language = {en} } @article{WaldholmWangBrodinetal.2011, author = {Waldholm, Johan and Wang, Zhi and Brodin, David and Tyagi, Anu and Yu, Simei and Theopold, Ulrich and {\"O}stlund Farrants, Ann Kristin and Visa, Neus}, title = {SWI/SNF regulates the alternative processing of a specific subset of pre-mRNAs in \(Drosophila\) \(melanogaster\)}, series = {BMC Molecular Biology}, volume = {12}, journal = {BMC Molecular Biology}, number = {46}, doi = {10.1186/1471-2199-12-46}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-142613}, pages = {1-12}, year = {2011}, abstract = {Background: The SWI/SNF chromatin remodeling factors have the ability to remodel nucleosomes and play essential roles in key developmental processes. SWI/SNF complexes contain one subunit with ATPase activity, which in Drosophila melanogaster is called Brahma (Brm). The regulatory activities of SWI/SNF have been attributed to its influence on chromatin structure and transcription regulation, but recent observations have revealed that the levels of Brm affect the relative abundances of transcripts that are formed by alternative splicing and/or polyadenylation of the same pre-mRNA. Results: We have investigated whether the function of Brm in pre-mRNA processing in Drosophila melanogaster is mediated by Brm alone or by the SWI/SNF complex. We have analyzed the effects of depleting individual SWI/SNF subunits on pre-mRNA processing throughout the genome, and we have identified a subset of transcripts that are affected by depletion of the SWI/SNF core subunits Brm, Snr1 or Mor. The fact that depletion of different subunits targets a subset of common transcripts suggests that the SWI/SNF complex is responsible for the effects observed on pre-mRNA processing when knocking down Brm. We have also depleted Brm in larvae and we have shown that the levels of SWI/SNF affect the pre-mRNA processing outcome in vivo. Conclusions: We have shown that SWI/SNF can modulate alternative pre-mRNA processing, not only in cultured cells but also in vivo. The effect is restricted to and specific for a subset of transcripts. Our results provide novel insights into the mechanisms by which SWI/SNF regulates transcript diversity and proteomic diversity in higher eukaryotes.}, language = {en} } @article{WagnerFischerThomaetal.2011, author = {Wagner, Toni U. and Fischer, Andreas and Thoma, Eva C. and Schartl, Manfred}, title = {CrossQuery : A Web Tool for Easy Associative Querying of Transcriptome Data}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-76088}, year = {2011}, abstract = {Enormous amounts of data are being generated by modern methods such as transcriptome or exome sequencing and microarray profiling. Primary analyses such as quality control, normalization, statistics and mapping are highly complex and need to be performed by specialists. Thereafter, results are handed back to biomedical researchers, who are then confronted with complicated data lists. For rather simple tasks like data filtering, sorting and cross-association there is a need for new tools which can be used by non-specialists. Here, we describe CrossQuery, a web tool that enables straight forward, simple syntax queries to be executed on transcriptome sequencing and microarray datasets. We provide deepsequencing data sets of stem cell lines derived from the model fish Medaka and microarray data of human endothelial cells. In the example datasets provided, mRNA expression levels, gene, transcript and sample identification numbers, GO-terms and gene descriptions can be freely correlated, filtered and sorted. Queries can be saved for later reuse and results can be exported to standard formats that allow copy-and-paste to all widespread data visualization tools such as Microsoft Excel. CrossQuery enables researchers to quickly and freely work with transcriptome and microarray data sets requiring only minimal computer skills. Furthermore, CrossQuery allows growing association of multiple datasets as long as at least one common point of correlated information, such as transcript identification numbers or GO-terms, is shared between samples. For advanced users, the object-oriented plug-in and event-driven code design of both server-side and client-side scripts allow easy addition of new features, data sources and data types.}, subject = {CrossQuery}, language = {en} } @article{WagnerFischerThomaetal.2011, author = {Wagner, Toni U. and Fischer, Andreas and Thoma, Eva C. and Schartl, Manfred}, title = {CrossQuery: A Web Tool for Easy Associative Querying of Transcriptome Data}, series = {PLoS ONE}, volume = {6}, journal = {PLoS ONE}, number = {12}, doi = {10.1371/journal.pone.0028990}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-134787}, pages = {e28990}, year = {2011}, abstract = {Enormous amounts of data are being generated by modern methods such as transcriptome or exome sequencing and microarray profiling. Primary analyses such as quality control, normalization, statistics and mapping are highly complex and need to be performed by specialists. Thereafter, results are handed back to biomedical researchers, who are then confronted with complicated data lists. For rather simple tasks like data filtering, sorting and cross-association there is a need for new tools which can be used by non-specialists. Here, we describe CrossQuery, a web tool that enables straight forward, simple syntax queries to be executed on transcriptome sequencing and microarray datasets. We provide deep-sequencing data sets of stem cell lines derived from the model fish Medaka and microarray data of human endothelial cells. In the example datasets provided, mRNA expression levels, gene, transcript and sample identification numbers, GO-terms and gene descriptions can be freely correlated, filtered and sorted. Queries can be saved for later reuse and results can be exported to standard formats that allow copy-and-paste to all widespread data visualization tools such as Microsoft Excel. CrossQuery enables researchers to quickly and freely work with transcriptome and microarray data sets requiring only minimal computer skills. Furthermore, CrossQuery allows growing association of multiple datasets as long as at least one common point of correlated information, such as transcript identification numbers or GO-terms, is shared between samples. For advanced users, the object-oriented plug-in and event-driven code design of both server-side and client-side scripts allow easy addition of new features, data sources and data types.}, language = {en} } @article{vonRahdenKircherLazariotouetal.2011, author = {von Rahden, Burkhard H.A. and Kircher, Stefan and Lazariotou, Maria and Reiber, Christoph and Stuermer, Luisa and Otto, Christoph and Germer, Christoph T. and Grimm, Martin}, title = {LgR5 expression and cancer stem cell hypothesis: clue to define the true origin of esophageal adenocarcinomas with and without Barrett's Esophagus?}, series = {Journal of Experimental \& Clinical Cancer Research}, volume = {30}, journal = {Journal of Experimental \& Clinical Cancer Research}, number = {23}, doi = {10.1186/1756-9966-30-23}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-137783}, year = {2011}, abstract = {Background Investigation of the expression of an intestinal stem cell marker in esophageal adenocarcinomas (EAC) with and without Barrett's Esophagus (BE), with respect to a cancer stem cell (CSC) hypothesis. Materials and methods Expression of a putative intestinal stem cell marker LgR5 was analyzed in esophageal cancer specimen (n = 70: 41 EAC with BE, 19 EAC without BE, and n = 10 esophageal squamous-cell carcinomas, ESCC) and in the adenocarcinoma cell line OE-33. Ki-67 and Cdx-2 were co-labelled with LgR5 in double staining experiments. Immunhistochemical expression results were confirmed by RT-PCR and correlated with tumor stage and five-year survival rates. Results LgR5was found expressed in 35 of 41 (85\%) EAC with BE and in 16 of 19 (81\%) EAC without BE. By contrast, LgR5 was not found to be expressed in ESCC. Quantification of immunolabeling showed 15\% LgR5+ cells in EAC with BE, 32\% LgR5+ cells in adjacent BE and 13\% in EAC without BE. Immunofluorescence double staining experiments with LgR5 and Ki-67 revealed a subpopulation (~5\%) of proliferating LgR+/Ki-67+ cells. On mRNA-level, expression of LgR5 was higher in BE in comparison to EAC (p = 0.0159). High levels of LgR5 expression in BE associated EAC were associated with poorer survival in univariate analysis. Conclusion The stem cell marker LgR5 is expressed in EAC, irrespective of association with BE, and appears to have negative impact on survival. The subset of proliferating LgR5+ cells (<5\%) might resemble rapidly cycling CSCs, which needs to be substantiated in further investigations.}, language = {en} } @article{vonRahdenKircherLazariotouetal.2011, author = {von Rahden, Burkhard H. A. and Kircher, Stefan and Lazariotou, Maria and Reiber, Christoph and Stuermer, Luisa and Otto, Christoph and Germer, Christoph T. and Grimm, Martin}, title = {LgR5 expression and cancer stem cell hypothesis: clue to define the true origin of esophageal adenocarcinomas with and without Barrett's Esophagus?}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-68810}, year = {2011}, abstract = {Background: Investigation of the expression of an intestinal stem cell marker in esophageal adenocarcinomas (EAC) with and without Barrett's Esophagus (BE), with respect to a cancer stem cell (CSC) hypothesis. Materials and methods: Expression of a putative intestinal stem cell marker LgR5 was analyzed in esophageal cancer specimen (n = 70: 41 EAC with BE, 19 EAC without BE, and n = 10 esophageal squamous-cell carcinomas, ESCC) and in the adenocarcinoma cell line OE-33. Ki-67 and Cdx-2 were co-labelled with LgR5 in double staining experiments. Immunhistochemical expression results were confirmed by RT-PCR and correlated with tumor stage and five-year survival rates. Results: LgR5was found expressed in 35 of 41 (85\%) EAC with BE and in 16 of 19 (81\%) EAC without BE. By contrast, LgR5 was not found to be expressed in ESCC. Quantification of immunolabeling showed 15\% LgR5+ cells in EAC with BE, 32\% LgR5+ cells in adjacent BE and 13\% in EAC without BE. Immunofluorescence double staining experiments with LgR5 and Ki-67 revealed a subpopulation (~5\%) of proliferating LgR+/Ki-67+ cells. On mRNAlevel, expression of LgR5 was higher in BE in comparison to EAC (p = 0.0159). High levels of LgR5 expression in BE associated EAC were associated with poorer survival in univariate analysis. Conclusion: The stem cell marker LgR5 is expressed in EAC, irrespective of association with BE, and appears to have negative impact on survival. The subset of proliferating LgR5+ cells (<5\%) might resemble rapidly cycling CSCs, which needs to be substantiated in further investigations.}, subject = {Medizin}, language = {en} } @article{vonKriesWeissFalkenhorstetal.2011, author = {von Kries, R{\"u}diger and Weiss, Susanne and Falkenhorst, Gerhard and Wirth, Stephan and Kaiser, Petra and Huppertz, Hans-Iko and Tenenbaum, Tobias and Schroten, Horst and Streng, Andrea and Liese, Johannes and Shai, Sonu and Niehues, Tim and Girschick, Hermann and Kuscher, Ellen and Sauerbrey, Axel and Peters, Jochen and Wirsing von Koenig, Carl Heinz and R{\"u}ckinger, Simon and Hampl, Walter and Michel, Detlef and Mertens, Thomas}, title = {Post-Pandemic Seroprevalence of Pandemic Influenza A (H1N1) 2009 Infection (Swine Flu) among Children < 18 Years in Germany}, series = {PLoS ONE}, volume = {6}, journal = {PLoS ONE}, number = {9}, doi = {10.1371/journal.pone.0023955}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-141698}, pages = {e23955}, year = {2011}, abstract = {Background: We determined antibodies to the pandemic influenza A (H1N1) 2009 virus in children to assess: the incidence of (H1N1) 2009 infections in the 2009/2010 season in Germany, the proportion of subclinical infections and to compare titers in vaccinated and infected children. Methodology/Principal Findings: Eight pediatric hospitals distributed over Germany prospectively provided sera from in-or outpatients aged 1 to 17 years from April 1(st) to July 31(st) 2010. Vaccination history, recall of infections and sociodemographic factors were ascertained. Antibody titers were measured with a sensitive and specific in-house hemagglutination inhibition test (HIT) and compared to age-matched sera collected during 6 months before the onset of the pandemic in Germany. We analyzed 1420 post-pandemic and 300 pre-pandemic sera. Among unvaccinated children aged 1-4 and 5-17 years the prevalence of HI titers (>= 1:10) was 27.1\% (95\% CI: 23.5-31.3) and 53.5\% (95\% CI: 50.9-56.2) compared to 1.7\% and 5.5\%, respectively, for pre-pandemic sera, accounting for a serologically determined incidence of influenza A (H1N1) 2009 during the season 2009/2010 of 25,4\% (95\% CI : 19.3-30.5) in children aged 1-4 years and 48.0\% (95\% CI: 42.6-52.0) in 5-17 year old children. Of children with HI titers >= 1: 10, 25.5\% (95\% CI: 22.5-28.8) reported no history of any infectious disease since June 2009. Among vaccinated children, 92\% (95\%-CI: 87.0-96.6) of the 5-17 year old but only 47.8\% (95\%-CI: 33.5-66.5) of the 1-4 year old children exhibited HI titers against influenza A virus (H1N1) 2009. Conclusion: Serologically determined incidence of influenza A (H1N1) 2009 infections in children indicates high infection rates with older children (5-17 years) infected twice as often as younger children. In about a quarter of the children with HI titers after the season 2009/2010 subclinical infections must be assumed. Low HI titers in young children after vaccination with the AS03(B)-adjuvanted split virion vaccine need further scrutiny.}, language = {en} } @article{vonBuerenOehlerShalabyetal.2011, author = {von Bueren, Andr{\´e} O. and Oehler, Christoph and Shalaby, Tarek and von Hoff, Katja and Pruschy, Martin and Seifert, Burkhardt and Gerber, Nicolas U. and Warmuth-Metz, Monika and Stearns, Duncan and Eberhart, Charles G. and Kortmann, Rolf D. and Rutkowski, Stefan and Grotzer, Michael A.}, title = {c-MYC expression sensitizes medulloblastoma cells to radio- and chemotherapy and has no impact on response in medulloblastoma patients}, series = {BMC Cancer}, volume = {11}, journal = {BMC Cancer}, number = {74}, doi = {10.1186/1471-2407-11-74}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-134185}, pages = {1-11}, year = {2011}, abstract = {Background: To study whether and how c-MYC expression determines response to radio-and chemotherapy in childhood medulloblastoma (MB). Methods: We used DAOY and UW228 human MB cells engineered to stably express different levels of c-MYC, and tested whether c-MYC expression has an effect on radio-and chemosensitivity using the colorimetric 3-(4,5-dimethylthiazol- 2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt (MTS) assay, clonogenic survival, apoptosis assays, cell cycle analysis, and western blot assessment. In an effort to validate our results, we analyzed c-MYC mRNA expression in formalin-fixed paraffin-embedded tumor samples from well-documented patients with postoperative residual tumor and compared c-MYC mRNA expression with response to radio-and chemotherapy as examined by neuroradiological imaging. Results: In DAOY -and to a lesser extent in UW228 -cells expressing high levels of c-MYC, the cytotoxicity of cisplatin, and etoposide was significantly higher when compared with DAOY/UW228 cells expressing low levels of c-MYC. Irradiation-and chemotherapy-induced apoptotic cell death was enhanced in DAOY cells expressing high levels of c-MYC. The response of 62 of 66 residual tumors was evaluable and response to postoperative radio-(14 responders (CR, PR) vs. 5 non-responders (SD, PD)) or chemotherapy (23 CR/PR vs. 20 SD/PD) was assessed. c-MYC mRNA expression was similar in primary MB samples of responders and non-responders (Mann-Whitney U test, p = 0.50, ratio 0.49, 95\% CI 0.008-30.0 and p = 0.67, ratio 1.8, 95\% CI 0.14-23.5, respectively). Conclusions: c-MYC sensitizes MB cells to some anti-cancer treatments in vitro. As we failed to show evidence for such an effect on postoperative residual tumors when analyzed by imaging, additional investigations in xenografts and larger MB cohorts may help to define the exact function of c-MYC in modulating response to treatment.}, language = {en} } @phdthesis{Vogl2011, author = {Vogl, Silvia}, title = {Investigation of individual differences in the metabolic elimination of drugs by the polymorphic enzymes CYP2C9, 2C19 and 2D6 based on metabolite profiling by LC-MS/MS}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-67216}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2011}, abstract = {Mit der vorliegenden Studie sollte zu dem wichtigen Forschungsfeld der Pharmakogenetik beigetragen werden, indem zum einen eine einfache und sichere kombinierte Ph{\"a}notypisierung der drei zuvor erw{\"a}hnten CYPs (CYP2D6, CYP2C9 und CYP2C19) entwickelt, und zum anderen die Vorhersagekraft des Genotyps f{\"u}r den gemessenen Ph{\"a}notyp n{\"a}her untersucht werden sollte. Es ist uns gelungen eine sichere, einfache, schnelle und kombinierte Ph{\"a}notypisierung der beiden wichtigen Monooxygenasen CYP2D6 und CYP2C9 zu etablieren. Zun{\"a}chst wurden dazu Wechselwirkungsstudien mit den ausgew{\"a}hlten Testsubstanzen Dextromethorphan (DEX, CYP2D6), Flurbiprofen (FLB, CYP2C9) und Omeprazole (OME, CYP2C19) durchgef{\"u}hrt. Es konnte gezeigt werden, dass DEX und FLB als Kombination verabreicht werden k{\"o}nnen. Die Gabe von OME gemeinsam mit FLB ver{\"a}ndert jedoch das Ergebnis der CYP2C9 Ph{\"a}notypisierung. Dies ist eine neue Erkenntnis, denn noch 2004 wurde ein Ph{\"a}notypisierungscocktail ver{\"o}ffentlicht, der die Kombination von FLB und OME enthielt. Bei der genannten Studie wurden jedoch, unseres Wissens nach, keine Wechselwirkungsstudien zu den einzelnen Testsubstanz-Kombinationen durchgef{\"u}hrt. Die von uns entwickelte Ph{\"a}notypisierungsmethode wurde durch Wechselwirkungsstudien verifiziert. Sie ist jedoch auch in anderen Bereichen den bisher ver{\"o}ffentlichten ph{\"a}notypisierungscocktails {\"u}berlegen. Zum einen wurden nur sehr kleine Dosen sicherer Testsubstanzen verwendet. Dies wurde durch Entwicklung neuer, sensitiver LC-MS/MS Methoden erm{\"o}glicht. Zum anderen ist diese neue Prozedur schnell und nicht-invasiv durchf{\"u}hrbar. Nach Verabreichung der Testsubstanz muss der Urin nur f{\"u}r zwei Stunden gesammelt werden. Zudem weisen unsere Ergebnisse darauf hin, dass die normalerweise durchgef{\"u}hrte, aufwendige Glucuronidspaltung des CYP2D6 abh{\"a}ngigen DEX-Metaboliten, Dextrorphan, vermutlich vernachl{\"a}ssigt werden kann. Die wichtigsten Ergebnisse dieser Studie sind jedoch die Einblicke, die in die Vorhersagekraft der CYP2D6 und CYP2C9 Genotypen f{\"u}r die entsprechenden Ph{\"a}notypen gewonnen werden konnten. Fast 300 ph{\"a}notypisierte Kaukasier wurden auch in Hinsicht auf die wichtigsten varianten Allele von CYP2D6, CYP2C9 und CYP2C19 mithilfe bekannter und neu etablierter Methoden genotypisiert. Aufgrund der parallelen Ph{\"a}no- und Genotypisierung konnten Geno- und Ph{\"a}notyp direkt korreliert werden. Mit linearen Modellen war es m{\"o}glich, allen detektierten varianten CYP2D6- und CYP2C9-Allelen Aktivit{\"a}tskoeffizienten zuzuweisen. Diese k{\"o}nnen nun verwendet werden, um den Beitrag der einzelnen Allele zur resultierenden Enzymaktivit{\"a}t zu bestimmen, wodurch sich die Vorhersage dieser Aktivit{\"a}t ausgehend vom Genotyp verbessern lassen sollte. Besonders f{\"u}r CYP2D6 erm{\"o}glicht das neue Korrelationsmodel pr{\"a}zisere Vorhersagen des Ph{\"a}notyps als bisher ver{\"o}ffentlichte Modelle. Zusammengefasst leistet diese Studie durch die Entwicklung eines sicheren und einfachen Ph{\"a}notypisierungsprozesses f{\"u}r CYP2D6 und CYP2C9 und durch die Bestimmung von Aktivit{\"a}tskoeffizienten f{\"u}r alle einbezogenen CYP2D6 und CYP2C9 Allele und der damit verbundenen pr{\"a}ziseren Vorhersage des Ph{\"a}notyps ausgehend vom Genotyp einen wesentlichen Beitrag zum Forschungsfeld der Pharmakogenetik.}, subject = {Pharmakogenetik}, language = {en} } @article{VogelLoeschbergerSaueretal.2011, author = {Vogel, Benjamin and L{\"o}schberger, Anna and Sauer, Markus and Hock, Robert}, title = {Cross-linking of DNA through HMGA1 suggests a DNA scaffold}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-68865}, year = {2011}, abstract = {Binding of proteins to DNA is usually considered 1D with one protein bound to one DNA molecule. In principle, proteins with multiple DNA binding domains could also bind to and thereby cross-link different DNA molecules. We have investigated this possibility using high-mobility group A1 (HMGA1) proteins, which are architectural elements of chromatin and are involved in the regulation of multiple DNA-dependent processes. Using direct stochastic optical reconstruction microscopy (dSTORM), we could show that overexpression of HMGA1a-eGFP in Cos-7 cells leads to chromatin aggregation. To investigate if HMGA1a is directly responsible for this chromatin compaction we developed a DNA cross-linking assay. We were able to show for the first time that HMGA1a can cross-link DNA directly. Detailed analysis using point mutated proteins revealed a novel DNA cross-linking domain. Electron microscopy indicates that HMGA1 proteins are able to create DNA loops and supercoils in linearized DNA confirming the cross-linking ability of HMGA1a. This capacity has profound implications for the spatial organization of DNA in the cell nucleus and suggests cross-linking activities for additional nuclear proteins.}, subject = {DNA}, language = {en} } @phdthesis{Veryha2011, author = {Veryha, Katarzyna}, title = {Qualitative and quantitative SEM margin analysis of Ormocer restorations in molars and premolars - 4 year long observation}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-64858}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2011}, abstract = {The most important aim of restorative therapy in dentistry is to achieve a restoration that remains dense from bacteria and this way from tooth pulp irritation as well. Patients on the other hand appreciate and expect additionally good aesthetics. This way the decision which material the practitioner should chose very often still causes dilemmas. The aim of this 4 year long study was to evaluate the Admira filling material, that belongs to ormocer group and its future in the area of restorative dentistry. SEM analysis of fillings margins followed on epoxy resin casts (achieved from impressions taken at each of the control appointments) and showed that after four years of clinical observation more than 90 percent of the restoratives margins remained perfectly adapted. Due to technical reasons the examination followed only in the enamel area and as a result this study is not answering the question of margin quality within the dentin.}, subject = {Ormocer}, language = {en} } @article{vanOorschotBeckmannSchulzeetal.2011, author = {van Oorschot, Birgitt and Beckmann, Gabriele and Schulze, Wolfgang and Rades, Dirk and Feyer, Petra}, title = {Radiotherapeutic options for symptom control in breast cancer}, series = {Breast Care}, volume = {6}, journal = {Breast Care}, number = {1}, issn = {1661-3791}, doi = {10.1159/000324564}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-199105}, pages = {14-19}, year = {2011}, abstract = {The majority of breast cancer patients will require radiation therapy at some time during the course of their disease. An estimated 30-50\% of all radiation treatments are of palliative nature, either to alleviate symptoms or prophylactic to prevent deterioration of quality of life due to locally progressive disease. Radiotherapy is a locally effective tool, and typically causes no systemic and mostly mild acute side effects. The following article provides an overview of options and decision-making in palliative radiotherapy for symptom control.}, language = {en} } @article{VandenHoveJakobSchrautetal.2011, author = {Van den Hove, Daniel and Jakob, Sissi Brigitte and Schraut, Karla-Gerlinde and Kenis, Gunter and Schmitt, Angelika Gertrud and Kneitz, Susanne and Scholz, Claus-J{\"u}rgen and Wiescholleck, Valentina and Ortega, Gabriela and Prickaerts, Jos and Steinbusch, Harry and Lesch, Klaus-Peter}, title = {Differential Effects of Prenatal Stress in 5-Htt Deficient Mice: Towards Molecular Mechanisms of Gene x Environment Interactions}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-75795}, year = {2011}, abstract = {Prenatal stress (PS) has been shown to influence the development of the fetal brain and to increase the risk for the development of psychiatric disorders in later life. Furthermore, the variation of human serotonin transporter (5-HTT, SLC6A4) gene was suggested to exert a modulating effect on the association between early life stress and the risk for depression. In the present study, we used a 5-Htt6PS paradigm to investigate whether the effects of PS are dependent on the 5-Htt genotype. For this purpose, the effects of PS on cognition, anxiety- and depression-related behavior were examined using a maternal restraint stress paradigm of PS in C57BL6 wild-type (WT) and heterozygous 5-Htt deficient (5-Htt +/2) mice. Additionally, in female offspring, a genome-wide hippocampal gene expression profiling was performed using the Affymetrix GeneChipH Mouse Genome 430 2.0 Array. 5-Htt +/2 offspring showed enhanced memory performance and signs of reduced anxiety as compared to WT offspring. In contrast, exposure of 5-Htt +/2 mice to PS was associated with increased depressive-like behavior, an effect that tended to be more pronounced in female offspring. Further, 5-Htt genotype, PS and their interaction differentially affected the expression of numerous genes and related pathways within the female hippocampus. Specifically, MAPK and neurotrophin signaling were regulated by both the 5-Htt +/2 genotype and PS exposure, whereas cytokine and Wnt signaling were affected in a 5-Htt genotype6PS manner, indicating a gene6environment interaction at the molecular level. In conclusion, our data suggest that although the 5-Htt +/2 genotype shows clear adaptive capacity, 5-Htt +/2 mice -particularly females- at the same time appear to be more vulnerable to developmental stress exposure when compared to WT offspring. Moreover, hippocampal gene expression profiles suggest that distinct molecular mechanisms mediate the behavioral effects of the 5-Htt genotype, PS exposure, and their interaction.}, subject = {Medizin}, language = {en} } @article{VandenHoveJakobSchrautetal.2011, author = {Van den Hove, Daniel and Jakob, Sissi Brigitte and Schraut, Karla-Gerlinde and Kenis, Gunter and Schmitt, Angelika Gertrud and Kneitz, Susanne and Scholz, Claus-J{\"u}rgen and Wiescholleck, Valentina and Ortega, Gabriela and Prickaerts, Jos and Steinbusch, Harry and Lesch, Klaus-Peter}, title = {Differential Effects of Prenatal Stress in 5-Htt Deficient Mice: Towards Molecular Mechanisms of Gene x Environment Interactions}, series = {PLoS ONE}, volume = {6}, journal = {PLoS ONE}, number = {8}, doi = {10.1371/journal.pone.0022715}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-135111}, pages = {e22715}, year = {2011}, abstract = {Prenatal stress (PS) has been shown to influence the development of the fetal brain and to increase the risk for the development of psychiatric disorders in later life. Furthermore, the variation of human serotonin transporter (5-HTT, SLC6A4) gene was suggested to exert a modulating effect on the association between early life stress and the risk for depression. In the present study, we used a 5-HttxPS paradigm to investigate whether the effects of PS are dependent on the 5-Htt genotype. For this purpose, the effects of PS on cognition, anxiety-and depression-related behavior were examined using a maternal restraint stress paradigm of PS in C57BL6 wild-type (WT) and heterozygous 5-Htt deficient (5-Htt +/-) mice. Additionally, in female offspring, a genome-wide hippocampal gene expression profiling was performed using the Affymetrix GeneChip (R) Mouse Genome 430 2.0 Array. 5-Htt +/- offspring showed enhanced memory performance and signs of reduced anxiety as compared to WT offspring. In contrast, exposure of 5-Htt +/- mice to PS was associated with increased depressive-like behavior, an effect that tended to be more pronounced in female offspring. Further, 5-Htt genotype, PS and their interaction differentially affected the expression of numerous genes and related pathways within the female hippocampus. Specifically, MAPK and neurotrophin signaling were regulated by both the 5-Htt +/- genotype and PS exposure, whereas cytokine and Wnt signaling were affected in a 5-Htt genotypexPS manner, indicating a genexenvironment interaction at the molecular level. In conclusion, our data suggest that although the 5-Htt +/- genotype shows clear adaptive capacity, 5-Htt +/- mice -particularly females-at the same time appear to be more vulnerable to developmental stress exposure when compared to WT offspring. Moreover, hippocampal gene expression profiles suggest that distinct molecular mechanisms mediate the behavioral effects of the 5-Htt genotype, PS exposure, and their interaction.}, language = {en} } @article{UppaluriNaglerStellamannsetal.2011, author = {Uppaluri, Sravanti and Nagler, Jan and Stellamanns, Eric and Heddergott, Niko and Herminghaus, Stephan and Pfohl, Thomas and Engstler, Markus}, title = {Impact of Microscopic Motility on the Swimming Behavior of Parasites: Straighter Trypanosomes are More Directional}, series = {PLoS Computational Biology}, volume = {7}, journal = {PLoS Computational Biology}, number = {6}, doi = {10.1371/journal.pcbi.1002058}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-140814}, pages = {e1002058}, year = {2011}, abstract = {Microorganisms, particularly parasites, have developed sophisticated swimming mechanisms to cope with a varied range of environments. African Trypanosomes, causative agents of fatal illness in humans and animals, use an insect vector (the Tsetse fly) to infect mammals, involving many developmental changes in which cell motility is of prime importance. Our studies reveal that differences in cell body shape are correlated with a diverse range of cell behaviors contributing to the directional motion of the cell. Straighter cells swim more directionally while cells that exhibit little net displacement appear to be more bent. Initiation of cell division, beginning with the emergence of a second flagellum at the base, correlates to directional persistence. Cell trajectory and rapid body fluctuation correlation analysis uncovers two characteristic relaxation times: a short relaxation time due to strong body distortions in the range of 20 to 80 ms and a longer time associated with the persistence in average swimming direction in the order of 15 seconds. Different motility modes, possibly resulting from varying body stiffness, could be of consequence for host invasion during distinct infective stages.}, language = {en} } @phdthesis{Tran2011, author = {Tran, Nam Binh}, title = {Climate change assessment in Southeast Asia and implications for agricultural production in Vietnam}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-64570}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2011}, abstract = {Seit vielen Jahren ist die Erforschung von Klimawandel und -schwankungen das zentrale Thema der Klimatologie. Besonderes deutlich wird dies anhand der IPCC-Berichte, ebenso wie der zahlreichen Einzelstudien zur Entwicklung des Klimas auf unterschiedlichsten raum-zeitlichen Skalen. Insbesondere seit den 1980er Jahren befassen sich zahlreiche Forschungsgruppen weltweit mit der systematischen Sammlung, Aufbereitung und auch Auswertung von Klimadaten. Diese Datengrundlage erlaubt Analysen zur Entwicklung der globalen Lufttemperatur, des Niederschlags und anderer Klimaelemente (Jones et al., 1986; Hansen und Lebedeff, 1987; Vinnikov et al., 1987, 1990). Das wichtigste {\"u}bergreifende Ergebnis dieser Untersuchungen ist die Feststellung einer globalen Erw{\"a}rmung w{\"a}hrend des 20. Jahrhunderts, die sich in den beiden letzten Jahrzehnten besonders intensivierte. Absch{\"a}tzungen {\"u}ber die Art und St{\"a}rke des Klimawandels auf gr{\"o}ßeren, planungsrelevanten Massst{\"a}ben sind jedoch nach wie vor mit großen Unsicherheiten verbunden. F{\"u}r eine detailliertere Erforschung der Auswirkungen der globalen Erw{\"a}rmung auf regionaler oder gar lokaler Ebene besteht daher noch großer Forschungsbedarf. In dieser Dissertation wird zu diesem Zweck ein statistischer Ansatz verfolgt. Dieser erlaubt die Identifikation systematischer Unterschiede zwischen den Auspr{\"a}gungen klimatologischer Feldgr{\"o}ßen (bodennahe Lufttemperatur und Niederschlag) wie sie von sogenannten General Circulation Models (GCMs) simuliert werden im Vergleich zu den betreffenden Parametern aus Beobachtungsdaten. Als Beobachtungsdaten werden die NCEP Reanalysen, die statistisch interpolierten Datens{\"a}tze der CRU sowie Stationsdaten aus Vietnam verwendet. Hierbei zeigt sich, dass die aktuellen Klimamodelle die r{\"a}umlichen Muster der betrachteten Variablen in befriedigender Weise reproduzieren. Die Analyse des regionalen Klimawandels in S{\"u}dost-Asien erfolgt durch die Auswertung von Klimamodellrechnungen. Diese wurden von verschiedenen GCMs durchgef{\"u}hrt, wobei unterschiedliche Annahmen {\"u}ber die zuk{\"u}nftigen Treibhausgasemissionen ber{\"u}cksichtigt wurden. Der Fokus dieser Dissertation ist die Analyse der projizierten zeitlichen Entwicklung von bodennaher Temperatur und Niederschlag im 21. Jahrhundert. Hierbei werden sowohl j{\"a}hrliche als auch saisonale Mittelwerte bzw. Summen ber{\"u}cksichtigt. Neben diesen rein physikalisch-klimatologischen Betrachtungen behandelt diese Dissertation auch einen angewandten Aspekt, n{\"a}mlich den Impakt des Klimawandels auf die Landwirtschaft, exemplarisch untersucht am Beispiel Vietnams. F{\"u}r die Absch{\"a}tzung der Vulnerabilit{\"a}t dieses essentiellen Wirtschaftsbereiches wird ein statistisches Modell entwickelt in das an klimatischen Parametern die bodennahe temperatur sowie der Niederschlag einfliessen. Diese Untersuchung leistet damit einen wichtigen Beitrag zum Wissenstand {\"u}ber die Auswirkungen des Klimawandels in den niederen Breiten. Die sozio-{\"o}konomische Entwicklung jedes Staates der Erde wird von den Folgen des Klimawandels beeinflusst, allerdings variiert der Grad der Beeintr{\"a}chtigung erheblich. Vermutlich werden Entwicklungsl{\"a}nder wie Vietnam die Auswirkungen des Klimawandels besonders stark zu sp{\"u}ren bekommen. Die Ursachen f{\"u}r diese hohe Vulnerabilit{\"a}t liegen unter anderem in der Wirtschaftsstruktur: der allgemein hohe Stellenwert nat{\"u}rlicher Ressourcen und eine geringe Diversit{\"a}t verringern hier die M{\"o}glichkeiten zur Adaption an die beobachteten und projizierten Ver{\"a}nderungen. Die vorliegende Dissertation gliedert sich wie folgt: In Kapitel 1 stellt eine allgemeine Einf{\"u}hrung zur Thematik dar. Die Begriffe Klima und Klimawandel sowie einige {\"u}bliche Modelle zum Klimawandel, verbunden mit einer Abw{\"a}gung der spezifischen Vor- und Nachteile, werden erl{\"a}utert. Kapitel 2 besch{\"a}ftigt sich mit der Methodik. Hier werden die r{\"a}umliche Interpolation sowie die angewendeten explorativen und inferentiellen statistischen Verfahren diskutiert. Die Kapitel 3 und 4 beschreiben die Datengrundlage und die betrachtete Region. Im Kapitel 5 werden die Untersuchungsergebnisse dargelegt. In Kapitel 6 erfolgt die Abschlussbetrachtung und ein Ausblick auf die Zukunft. Am Ende der Dissertation finden sich die verwendeten Quellen sowie ein Appendix mit landwirtschaftlichen Daten.}, subject = {Klima{\"a}nderung}, language = {en} } @article{TonyBurmesterSchulzeKoopsetal.2011, author = {Tony, Hans-Peter and Burmester, Gerd and Schulze-Koops, Hendrik and Grunke, Mathias and Henes, Joerg and K{\"o}tter, Ina and Haas, Judith and Unger, Leonore and Lovric, Svjetlana and Haubitz, Marion and Fischer-Betz, Rebecca and Chehab, Gamal and Rubbert-Roth, Andrea and Specker, Christof and Weinerth, Jutta and Holle, Julia and M{\"u}ller-Ladner, Ulf and K{\"o}nig, Ramona and Fiehn, Christoph and Burgwinkel, Philip and Budde, Klemens and S{\"o}rensen, Helmut and Meurer, Michael and Aringer, Martin and Kieseier, Bernd and Erfurt-Berge, Cornelia and Sticherling, Michael and Veelken, Roland and Ziemann, Ulf and Strutz, Frank and von Wussow, Praxis and Meier, Florian MP and Hunzelmann, Nico and Schmidt, Enno and Bergner, Raoul and Schwarting, Andreas and Eming, R{\"u}diger and Schwarz-Eywill, Michael and Wassenberg, Siegfried and Fleck, Martin and Metzler, Claudia and Zettl, Uwe and Westphal, Jens and Heitmann, Stefan and Herzog, Anna L. and Wiendl, Heinz and Jakob, Waltraud and Schmidt, Elvira and Freivogel, Klaus and D{\"o}rner, Thomas and Hertl, Michael and Stadler, Rudolf}, title = {Safety and clinical outcomes of rituximab therapy in patients with different autoimmune diseases: experience from a national registry (GRAID)}, series = {Arthritis Research \& Therapy}, volume = {13}, journal = {Arthritis Research \& Therapy}, number = {R75}, doi = {10.1186/ar3337}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-142856}, pages = {1-14}, year = {2011}, abstract = {Introduction: Evidence from a number of open-label, uncontrolled studies has suggested that rituximab may benefit patients with autoimmune diseases who are refractory to standard-of-care. The objective of this study was to evaluate the safety and clinical outcomes of rituximab in several standard-of-care-refractory autoimmune diseases (within rheumatology, nephrology, dermatology and neurology) other than rheumatoid arthritis or non-Hodgkin's lymphoma in a real-life clinical setting. Methods: Patients who received rituximab having shown an inadequate response to standard-of-care had their safety and clinical outcomes data retrospectively analysed as part of the German Registry of Autoimmune Diseases. The main outcome measures were safety and clinical response, as judged at the discretion of the investigators. Results: A total of 370 patients (299 patient-years) with various autoimmune diseases (23.0\% with systemic lupus erythematosus, 15.7\% antineutrophil cytoplasmic antibody-associated granulomatous vasculitides, 15.1\% multiple sclerosis and 10.0\% pemphigus) from 42 centres received a mean dose of 2,440 mg of rituximab over a median (range) of 194 (180 to 1,407) days. The overall rate of serious infections was 5.3 per 100 patient-years during rituximab therapy. Opportunistic infections were infrequent across the whole study population, and mostly occurred in patients with systemic lupus erythematosus. There were 11 deaths (3.0\% of patients) after rituximab treatment (mean 11.6 months after first infusion, range 0.8 to 31.3 months), with most of the deaths caused by infections. Overall (n = 293), 13.3\% of patients showed no response, 45.1\% showed a partial response and 41.6\% showed a complete response. Responses were also reflected by reduced use of glucocorticoids and various immunosuppressives during rituximab therapy and follow-up compared with before rituximab. Rituximab generally had a positive effect on patient well-being (physician's visual analogue scale; mean improvement from baseline of 12.1 mm)}, language = {en} } @phdthesis{Tichy2011, author = {Tichy, Michael}, title = {On algebraic aggregation methods in additive preconditioning}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-56541}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2011}, abstract = {In the following dissertation we consider three preconditioners of algebraic multigrid type, though they are defined for arbitrary prolongation and restriction operators, we consider them in more detail for the aggregation method. The strengthened Cauchy-Schwarz inequality and the resulting angle between the spaces will be our main interests. In this context we will introduce some modifications. For the problem of the one-dimensional convection we obtain perfect theoretical results. Although this is not the case for more complex problems, the numerical results we present will show that the modifications are also useful in these situation. Additionally, we will consider a symmetric problem in the energy norm and present a simple rule for algebraic aggregation.}, subject = {Pr{\"a}konditionierung}, language = {en} } @phdthesis{Tichy2011, author = {Tichy, Diana}, title = {On the Fragility Index}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-73610}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2011}, abstract = {The Fragility Index captures the amount of risk in a stochastic system of arbitrary dimension. Its main mathematical tool is the asymptotic distribution of exceedance counts within the system which can be derived by use of multivariate extreme value theory. Thereby the basic assumption is that data comes from a distribution which lies in the domain of attraction of a multivariate extreme value distribution. The Fragility Index itself and its extension can serve as a quantitative measure for tail dependence in arbitrary dimensions. It is linked to the well known extremal index for stochastic processes as well the extremal coefficient of an extreme value distribution.}, subject = {Extremwertstatistik}, language = {en} } @phdthesis{Tian2011, author = {Tian, Rui}, title = {Structural and functional organization of synaptic proteins in Drosophila melanogaster}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-57399}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2011}, abstract = {Structural and functional modifications of synaptic connections ("synaptic plasticity") are believed to mediate learning and memory processes. Thus, molecular mechanisms of how synapses assemble in both structural and functional terms are relevant for our understanding of neuronal development as well as the processes of learning and memory. Synapses form by an asymmetric association of highly specialized membrane domains: at the presynaptic active zone transmitter filled vesicles fuse, while transmitter receptors at the opposite postsynaptic density sense this signal. By genetic analysis, matrix proteins of active zones from various families have been shown to be important for fast vesicle fusion, and were suggested to contribute to synapse stability and assembly. The Sigrist lab in collaboration with the Buchner lab previously had shown that the large scaffold protein Bruchpilot (Brp) is essential for both the structural and functional integrity of active zones and for synaptic plasticity in Drosophila melanogaster. The work described in this thesis investigated several candidate proteins which appear to be involved in preand postsynaptic function, as summarized in the following: (1) DREP-2 (DEF45 related protein-2) had been found by co-immunoprecipitations with anti-Brp antibodies by Dr. Manuela Schmidt (unpublished data). Mutants and antibodies for the further study of DREP- 2 were generated in this thesis. Yeast two hybrid results suggest that DREP-2 might interact with dynein light chain 2, while in vivo imaging indicates that DREP-2 might be involved in bidirectional axonal transport. (2) Coimmunoprecipitation and pull down experiments suggested that the ARFGAP [ADP-ribosylation factor (ARF)-directed GTPase activating protein (GAP)] protein GIT (G-protein coupled receptor kinase interacting protein) could interact with the endocytosis associated molecule Stoned B (StnB). Mutants in the dgit gene showed an accumulation of large size vesicles, membrane intermediates and decreased vesicle density at the 3rd instar larval neuromuscular junction (NMJ) by electron microscopy (EM). The phenotypes accumulation of large size vesicles and membrane intermediates could be rescued partially by expression of Drosophila GIT (DGIT) or human GIT in dgit mutant background. Furthermore, by immunofluorescence the dgit mutant shows specifically decreased levels of StnB, which could be restored partially by the expression of DGIT. These results strongly support the suggestion that DGIT interacts with StnB, which is involved in the regulation of vesicle size, endocytosis or recycling of synaptic vesicles (SVs). Furthermore, the dgit mutants also showed signs of a mislocalization of the presynaptic protein Brp relative to the postsynaptic protein GluRIID, which could be rescued by expression of DGIT or human GIT in the dgit mutant background, but not by StnB. These results suggest that GIT on one hand executes roles in the regulation of synaptic vesicle endocytosis, but potentially also has structural roles for synapse assembly (3) Djm-1 is a candidate locus to mediate mental retardation in human patients when it is mutated. As a first step towards an understanding of the mechanistic role of DJM-1, Drosophila genetics were used to address DJM-1 function. So far, however, the djm-1 mutant generated in this thesis did not show a nervous system phenotype.}, subject = {Taufliege}, language = {en} } @article{ThormannRaupachWagneretal.2011, author = {Thormann, Birthe and Raupach, Michael J. and Wagner, Thomas and W{\"a}gele, Johann W. and Peters, Marcell K.}, title = {Testing a Short Nuclear Marker for Inferring Staphylinid Beetle Diversity in an African Tropical Rain Forest}, series = {PLoS ONE}, volume = {6}, journal = {PLoS ONE}, number = {3}, doi = {10.1371/journal.pone.0018101}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-142666}, pages = {e18101}, year = {2011}, abstract = {Background: The use of DNA based methods for assessing biodiversity has become increasingly common during the last years. Especially in speciose biomes as tropical rain forests and/or in hyperdiverse or understudied taxa they may efficiently complement morphological approaches. The most successful molecular approach in this field is DNA barcoding based on cytochrome c oxidase I (COI) marker, but other markers are used as well. Whereas most studies aim at identifying or describing species, there are only few attempts to use DNA markers for inventorying all animal species found in environmental samples to describe variations of biodiversity patterns. Methodology/Principal Findings: In this study, an analysis of the nuclear D3 region of the 28S rRNA gene to delimit species-like units is compared to results based on distinction of morphospecies. Data derived from both approaches are used to assess diversity and composition of staphylinid beetle communities of a Guineo-Congolian rain forest in Kenya. Beetles were collected with a standardized sampling design across six transects in primary and secondary forests using pitfall traps. Sequences could be obtained of 99\% of all individuals. In total, 76 molecular operational taxonomic units (MOTUs) were found in contrast to 70 discernible morphospecies. Despite this difference both approaches revealed highly similar biodiversity patterns, with species richness being equal in primary and secondary forests, but with divergent species communities in different habitats. The D3-MOTU approach proved to be an efficient tool for biodiversity analyses. Conclusions/Significance: Our data illustrate that the use of MOTUs as a proxy for species can provide an alternative to morphospecies identification for the analysis of changes in community structure of hyperdiverse insect taxa. The efficient amplification of the D3-marker and the ability of the D3-MOTUs to reveal similar biodiversity patterns as analyses of morphospecies recommend its use in future molecular studies on biodiversity.}, language = {en} } @phdthesis{Thoma2011, author = {Thoma, Eva Christina}, title = {Directed differentiation of pluripotent stem cells induced by single genes}, url = {http://nbn-resolving.de/urn:nbn:de:bvb:20-opus-54706}, school = {Universit{\"a}t W{\"u}rzburg}, year = {2011}, abstract = {Pluripotency describes the ability of stem cells to form every cell type of the body.. Pluripotent stem cells are e.g. embryonic stem cells (ESCs), but also the so called induced pluripotent stem cells (IPS cells), that are generated by reprogramming differentiated somatic cells into a pluripotent state. Furthermore, it has been shown that spermatogonia (SG) derived from adult testes of mouse or human are pluripotent. Because of their ability to differentiate into every somatic cell type, pluripotent stem cells have a unique status in research and regenerative medicine. For the latter, they offer a valuable opportunity to replace destroyed tissues or organs. For basic research, stem cells represent a useful system to study differentiation or developmental processes that are difficult to access in the physiological situation e.g. during embryogenesis. Both applications, however, require methods that allow efficient and directed differentiation of stem cells into defined specialized cell types. This study first aims to investigate the differentiation potential of SG derived from the teleost fish medaka (Oryzias latipes). My results demonstrate that medaka SG are able to form different somatic cell types, namely adipocytes, melanocytes, osteoblasts, and neurons. This indicates that medake SG have retained a broad differentiation potential suggesting that pluripotency is not restricted to mouse and human SG but might be conserved among vertebrates. Next, I wanted to establish a differentiation method that is solely based on ectopic expression of genes known to be essential for the formation of certain somatic cell types - so called master regulators (MRs). My findings show that ectopic expression of the melanocyte-specific transcription factor mitf-m that has previously been shown to induce differentiation of medaka ESCs into pigment cells resulted in the formation of the same cell type in medaka SG. This approach could be used to generate other somatic cell types. Thus, ectopic expression of the MRs cbfa1 and mash1 in MF-SG was sufficient to induce differentiation into osteoblasts and neurons, respectively. Interestingly, these differentiation processes included the activation of genes that are expressed earlier during embryogenesis than the differentiation-inducing MR. Furthermore, my findings show that the approach of MR-induced differentiation can be transferred to mammalian stem cell systems. Ectopic expression of the neural transcription factor ngn2 was sufficient to induce efficient and rapid differentiation of neurons in mouse ESCs. This differentiation process also included the induction of genes that in vivo are activated at earlier stages that ngn2. By generating a transgenic cell line allowing induction of ectopic ngn2 expression, it was possible to obtain a relatively pure culture of functional neurons. Ngn2-induced differentiation did not require any additional signals and occurred even under pluripotency promoting conditions. Moreover, ectopic expression of ngn2 did also induce the formation of cells with neuronal morphology in IPS cells indicating that MR-induced differentiation is operative in different stem cell types. Furthermore, protein transduction of Ngn2 into mouse ESCs also resulted in a neuronal differentiation process up to the appearance of neural precursor cells. Last, my results show that MR-induced differentiation can also be used to generate other cell types than neurons from mouse ESCs. Myoblasts and macrophage-like cells were generated by ectopic expression of the MRs myoD and cebpa, respectively. Using transgenic cell lines enabling induction of MR expression it was possible to obtain mixed cultures with two different differentiation processes occurring in parallel. Altogether this study shows that ectopic expression of single genes is sufficient to induce directed differentiation of stem cells into defined cell types. The feasibility of this approach was demonstrated for different MRs and consequently different somatic cell types. Furthermore, MR induced differentiation was operative in different stem cell types from fish and mouse. Thus, one can conclude that certain genes are able to define cell fates in in vitro stem cell systems and that this cell fate defining potential appears to be a conserved feature in vertebrates. These findings therefore provide new insights in the role of MRs in cell commitment and differentiation processes. Furthermore, this study presents a new method to induce directed differentiation of stem cells that offers several advantages regarding efficiency, rapidness, and reproducibility. MR-induced differentiation therefore represents a promising tool for both stem cell research and regenerative medicine.}, subject = {Stammzelle}, language = {en} }