TY - JOUR A1 - Arrowsmith, Merle A1 - Böhnke, Julian A1 - Braunschweig, Holger A1 - Celik, Mehmet A1 - Claes, Christina A1 - Ewing, William A1 - Krummenacher, Ivo A1 - Lubitz, Katharina A1 - Schneider, Christoph T1 - Neutral Diboron Analogues of Archetypal Aromatic Species by Spontaneous Cycloaddition JF - Angewandte Chemie, International Edition N2 - Among the numerous routes organic chemists have developed to synthesize benzene derivatives and heteroaro- matic compounds, transition-metal-catalyzed cycloaddition reactions are the most elegant. In contrast, cycloaddition reactions of heavier alkene and alkyne analogues, though limited in scope, proceed uncatalyzed. In this work we present the first spontaneous cycloaddition reactions of lighter alkene and alkyne analogues. Selective addition of unactivated alkynes to boron–boron multiple bonds under ambient con- ditions yielded diborocarbon equivalents of simple aromatic hydrocarbons, including the first neutral 6 π-aromatic dibora- benzene compound, a 2  π-aromatic triplet biradical 1,3-dibor- ete, and a phosphine-stabilized 2  π-homoaromatic 1,3-dihydro- 1,3-diborete. DFT calculations suggest that all three com- pounds are aromatic and show frontier molecular orbitals matching those of the related aromatic hydrocarbons, C\(_6\)H\(_6\) and C\(_4\)H\(_4\)\(^{2+}\), and homoaromatic C\(_4\)H\(_5\)\(^+\). KW - Aromaticity KW - Biradicals KW - Boron KW - Cycloaddition KW - Multiple bonds Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-138226 N1 - This is the peer reviewed version of the following article: Angew. Chem. Int. Ed. 2016, 55, 11271–11275, which has been published in final form at 10.1002/anie.201602384. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving. VL - 55 ER - TY - JOUR A1 - Arrowsmith, Merle A1 - Böhnke, Julian A1 - Braunschweig, Holger A1 - Celik, Mehmet A1 - Dellermann, Theresa A1 - Hammond, Kai T1 - Uncatalyzed Hydrogenation of First-Row Main Group Multiple Bonds JF - Chemistry, A European Journal N2 - Room temperature hydrogenation of an SIDep-stabilized diboryne (SIDep = 1,3-bis(diethylphenyl)-4,5-dihydroimidazol-2-ylidene) and a CAAC-supported diboracumulene (CAAC = 1-(2,6- diisopropylphenyl)-3,3,5,5-tetramethylpyrrolidin-2-ylidene) provided the first selective route to the corresponding 1,2-dihydrodiborenes. DFT calculations showed an overall exothermic (ΔG = 19.4 kcal mol\(^{-1}\) two-step asynchronous H\(_2\) addition mechanism proceeding via a bridging hydride. KW - diborenes KW - carbenes KW - hydrogenation KW - main-group chemistry KW - reaction mechanism KW - Diborane Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-139364 N1 - This is the peer reviewed version of the following article: Chemistry, A European Journal, 2016, 22, 17169–17172, which has been published in final form at 10.1002/chem.201604094. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving. VL - 22 IS - 48 SP - 17169 EP - 17172 ER - TY - THES A1 - Asmus, Elisabeth T1 - Mode of Action of Adjuvants for Foliar Application T1 - Wirkmechanismen von Adjuvantien für die Blattflächenapplikation N2 - Adjuvants are compounds added to an agrochemical spray formulation to improve or modify the action of an active ingredient (AI) or the physico-chemical characteristics of the spray liquid. Adjuvants can have more than only one distinct mode of action (MoA) during the foliar spray application process and they are generally known to be the best tools to improve agrochemical formulations. The main objective for this work was to elucidate the basic MoA of adjuvants by uncoupling different aspects of the spray application. Laboratory experiments, beginning from retention and spreading characteristics, followed by humectant effects concerning the spray deposit on the leaf surface and ultimately the cuticular penetration of an AI, were figured out to evaluate overall in vivo effects of adjuvants which were also obtained in a greenhouse spray test. For this comprehensive study, the surfactant classes of non-ionic sorbitan esters (Span), polysorbates (Tween) and oleyl alcohol polyglycol ether (Genapol O) were generally considered because of their common promoting potential in agrochemical formulations and their structural diversity. The reduction of interfacial tension is one of the most crucial physico-chemical properties of surfactants. The dynamic surface tension (DST) was monitored to characterise the surface tension lowering behaviour which is known to influence the droplet formation and retention characteristics. The DST is a function of time and the critical time frame of droplet impact might be at about 100 ms. None of the selected surfactants were found to lower the surface tension sufficiently during this short timeframe (chapter I). At ca. 100 ms, Tween 20 resulted in the lowest DST value. When surfactant monomers are fully saturated at the droplet-air-interface, an equilibrium surface tension (STeq) value can be determined which may be used to predict spreading or run-off effects. The majority of selected surfactants resulted in a narrow distribution of STeq values, ranging between 30 and 45 mN m- 1. Nevertheless, all surfactants were able to decrease the surface tension considerably compared to pure water (72 mN m- 1). The influence of different surfactants on the wetting process was evaluated by studying time-dependent static contact angles on different surfaces and the droplet spread area on Triticum aestivum leaves after water evaporation. The spreading potential was observed to be better for Spans than for Tweens. Especially Span 20 showed maximum spreading results. To transfer laboratory findings to spray application, related to field conditions, retention and leaf coverage was measured quantitatively on wheat leaves by using a variable track sprayer. Since the retention process involves short time dynamics, it is well-known that the spray retention on a plant surface is not correlated to STeq but to DST values. The relationship between DST at ca. 100 ms and results from the track sprayer showed increasing retention results with decreasing DST, whereas at DST values below ca. 60 mN m- 1 no further retention improvement could be observed. Under field conditions, water evaporates from the droplet within a few seconds to minutes after droplet deposition on the leaf surface. Since precipitation of the AI must essentially being avoided by holding the AI in solution, so-called humectants are used as tank-mix adjuvants. The ability of pure surfactants to absorb water from the surrounding atmosphere was investigated comprehensively by analysing water sorption isotherms (chapter II). These isotherms showed an exponential shape with a steep water sorption increase starting at 60% to 70% RH. Water sorption was low for Spans and much more distinct for the polyethoxylated surfactants (Tweens and Genapol O series). The relationship between the water sorption behaviour and the molecular structure of surfactants was considered as the so-called humectant activity. With an increasing ethylene oxide (EO) content, the humectant activity increased concerning the particular class of Genapol O. However, it could be shown that the moisture absorption across all classes of selected surfactants correlates rather better with their hydrophilic-lipophilic balance values with the EO content. All aboveground organs of plants are covered by the cuticular membrane which is therefore the first rate limiting barrier for AI uptake. In vitro penetration experiments through an astomatous model cuticle were performed to study the effects of adjuvants on the penetration of the lipophilic herbicide Pinoxaden (PXD) (chapter III). In order to understand the influence of different adjuvant MoA like humectancy, experiments were performed under three different humidity levels. No explicit relationship could be found between humidity levels and the PXD penetration which might be explained by the fact that humidity effects would rather affect hydrophilic AIs than lipophilic ones. Especially for Tween 20, it became obvious that a complex balance between multiple MoA like spreading, humectancy and plasticising effects have to be considered. Greenhouse trials, focussing the adjuvant impact on in vivo action of PXD, were evaluated on five different grass-weed species (chapter III). Since agrochemical spray application and its following action on living plants also includes translocation processes in planta and species dependent physiological effects, this investigation may help to simulate the situation on the field. Even though the absolute weed damage was different, depending both on plant species and also on PXD rates, adjuvant effects in greenhouse experiments displayed the same ranking as in cuticular penetration studies: Tween 20 > Tween 80 > Span 20 ≥ Span 80. Thus, the present work shows for the first time that findings obtained in laboratory experiments can be successfully transferred to spray application studies on living plants concerning adjuvant MoA. A comparative analysis, using radar charts, could demonstrate systematic derivations from structural similarities of adjuvants to their MoA (summarising discussion and outlook). Exemplarily, Tween 20 and Tween 80 cover a wide range of selected variables by having no outstanding MoA improving one distinct process during foliar application, compared to non-ethoxylated Span 20 and Span 80 which primarily revealed a surface active action. Most adjuvants used in this study represent polydisperse mixtures bearing a complex distribution of EO and aliphatic chains. From this study it seems alike that adjuvants having a wide EO distribution offer broader potential than adjuvants with a small EO distribution. It might be a speculation that due to this broad distribution of single molecules, all bearing their individual specific physico-chemical nature, a wide range of properties concerning their MoA is covered. N2 - Adjuvantien sind chemische Verbindungen, die einer Pflanzenschutzformulierung hinzugefügt werden, um die Wirkung der Aktivsubstanz oder die physikalisch-chemischen Eigenschaften der Spritzbrühe zu verbessern oder zu modifizieren. Sie können mehr als nur einen einzigen bestimmten Wirkmechanismus während der Blattflächenapplikation aufweisen, sodass sie gemeinhin als wirksamste Hilfsmittel in Pflanzenschutzformulierungen benutzt werden. Der Schwerpunkt dieser Arbeit lag darauf, ihre wesentlichen Wirkmechanismen aufzuklären, indem verschiedene Aspekte der Applikation entkoppelt und unabhängig voneinander untersucht wurden. Hierzu wurden Laborversuche durchgeführt, beginnend mit dem Retentions- und Spreitungsverhalten, über die „humectant“- Eigenschaft, den Tropfenrückstand betreffend und schließlich die kutikuläre Penetration einer Aktivsubstanz. Um schlussendlich die ineinander übergreifenden in vivo Mechanismen von Adjuvantien zusammenfassend bewerten zu können, wurde zusätzlich ein Gewächshausprayversuch ausgeführt. Für diese mechanismenübergreifende Studie wurden aufgrund ihrer allgemein begünstigenden Eigenschaften in Formulierungen und ihrer strukturellen Vielfalt die Tensidklassen der Sorbitanester (Span), Polysorbate (Tween) und Oleylalkoholpolyglykolether (Genapol O) verwendet. Die Absenkung der Grenzflächenspannung ist eine der wesentlichen physikalisch-chemischen Eigenschaften von Tensiden. Die dynamische Oberflächenspannung (DST) wurde untersucht, um das Verhalten beim Absenken der Oberflächenspannung einzuschätzen, welches die Tropfenbildung und die Retentionseigenschaften beeinflusst. Die DST ist zeitabhängig und das kritische Zeitfenster, in dem ein Tropfen auf die Pflanzenoberfläche auftrifft, beläuft sich auf ca. 100 ms. Innerhalb dieses Zeitrahmens konnte keines von den ausgewählten Tensiden die Oberflächenspannung hinreichend herabsetzen (vgl. Kapitel I). Bei ca. 100 ms wies Tween 20 die niedrigsten DST-Werte auf. Bei vollständiger Absättigung der Tropfen-Luft-Grenzfläche durch Tensidmonomere kann eine statische Oberflächenspannung (STeq) bestimmt werden. Diese physikalische Größe kann benutzt werden, um Spreitungs- und „run-off“- Effekte abzuschätzen. Der Großteil der betrachteten Tenside zeigte ähnliche STeq-Ergebnisse zwischen 30 und 45 mN m- 1. Somit waren alle Tenside in der Lage, die Oberflächenspannung von Wasser (72 mN m- 1) beträchtlich abzusenken. Der Einfluss von Tensiden auf den Benetzungsprozess wurde sowohl mit Hilfe von zeitabhängigen, statischen Kontaktwinkelmessungen auf verschiedenen Oberflächen, als auch nach der Wasserverdunstung auf Basis der Spreitungsfläche der Tropfen auf Triticum aestivum Blättern analysiert. Dabei zeigte die Klasse der Spans, besonders Span 20, ein besseres Benetzungsverhalten als die Klasse der Tweens. Um die Erkenntnisse aus dem Labor auf die Sprayapplikation auf dem Feld zu übertragen, wurden Retention und Blattbedeckung quantitativ auf Weizenoberflächen mit Hilfe eines „tracksprayers“ bestimmt. Da der Retention ein sehr schnell ablaufender, dynamischer Prozess zugrunde liegt, korreliert sie nicht mit der statischen, sondern mit der dynamischen Oberflächenspannung. Die Beziehung zwischen DST, bei ca. 100 ms und den „tracksprayer“- Ergebnissen zeigte eine Retentionszunahme bei abnehmender DST. Dabei konnte keine weitere Retentionsverbesserung bei DST-Werten unterhalb von ca. 60 mN m- 1 erzielt werden. Nachdem der Tropfen auf der Blattoberfläche gelandet ist, verdunstet Wasser unter Feldbedingungen aus dem Sprühtropfen innerhalb weniger Sekunden bis Minuten. Da das Auskristallisieren der Aktivsubstanz zwingend vermieden werden muss, werden sog. „humectants“ (dt. Feuchthaltemittel) als Tankmix-Adjuvantien eingesetzt, um die Aktivsubstanz in Lösung zu halten. Die Fähigkeit von Tensiden, Wasser aus der umgebenden Atmosphäre zu binden, wurde mit Hilfe von Wassersorptionsisothermen umfassend analysiert (vgl. Kapitel II). Diese Isothermen zeigten einen exponentiellen Verlauf mit einem steilen Anstieg der Wassersorption, beginnend ab ca. 60 bis 70% RH (relative Luftfeuchte). Dabei zeigten Spans eine geringere Wassersorption als die polyethoxylierten Tenside (Tweens und Genapol O). Die Beziehung zwischen dem Wassersorptionsverhalten und der molekularen Struktur der Tenside wurde als sog. „humectant Aktivität“ betrachtet. Speziell für die Klasse der Genapol O Tenside, wurde mit zunehmenden Ethylenoxidgehalt (EO) eine Zunahme der „humectant Aktivität“ nachgewiesen. Es konnte jedoch auch gezeigt werden, dass die Wassersorption über alle Klassen der hier ausgewählten Tenside eher mit dem „hydrophilic-lipophilic-balance“- Wert (HLB) als mit dem EO-Gehalt korreliert. Die Kutikula ist eine Membran, die alle oberirdischen Pflanzenorgane bedeckt. Damit stellt sie die wichtigste transportlimitierende Barriere für die Aufnahme von Pflanzenschutzmittelwirkstoffen dar. Um die Wirkung von Adjuvantien auf die Penetration des lipophilen Herbizids Pinoxaden (PXD) zu bestimmen, wurden in vitro Penetrationsexperimente durch eine astomatäre Modellkutikula durchgeführt (vgl. Kapitel III). Um den Einfluss verschiedener Wirkmechanismen, wie z.B. die der „humectant“- Eigenschaft zu verstehen, wurden diese Versuche unter drei verschiedenen Luftfeuchtebedingungen durchgeführt. Hierbei konnte kein klarer Zusammenhang zwischen der relativen Luftfeuchte und der PXD-Penetration nachgewiesen werden. Eine Ursache dafür könnte sein, dass sich Luftfeuchteeffekte eher auf hydrophile als auf lipophile Stoffe auswirken. Vielmehr wurde deutlich, dass hier eine komplexe Kombination aus verschiedenen Wirkmechanismen, wie z.B. Spreitungs-, „humectant“- und Weichmachereffekte, zum Tragen kommt. Um den Einfluss von Adjuvantien auf die in vivo Wirkung von PXD zu analysieren, wurden Gewächshausstudien mit fünf verschiedenen Ungräsern durchgeführt (vgl. Kapitel III). Da die Applikation von Pflanzenschutzwirkstoffen auch deren nachfolgende Wirkung auf lebende Pflanzen, wie z.B. Translokationsprozesse in planta und speziesspezifische physiologische Effekte beinhaltet, kann diese Untersuchung helfen, die Situation auf dem Feld besser zu simulieren. Durch die sowohl verschiedenen Pflanzenspezies als auch PXD-Konzentrationen variierte die absolute Schädigung der Ungräser stark. Dennoch kam es zur gleichen Reihenfolge der Auswirkung der Adjuvantien wie in den in vitro Kutikula-Penetrationsversuchen: Tween 20 > Tween 80 > Span 20 ≥ Span 80. Somit konnte in der vorliegenden Arbeit zum ersten Mal gezeigt werden, dass Erkenntnisse aus Laborversuchen, die die Wirkmechanismen von Adjuvantien betreffen, erfolgreich auf die Sprayapplikation auf Pflanzen übertragen werden können. Um eine systematische Herleitung der Wirkmechanismen von Adjuvantien abschließend zusammenzufassen, vergleichen und bewerten zu können, wurde dies mit Hilfe von Netzdiagrammen grafisch dargestellt (vgl. Zusammenfassende Diskussion und Ausblick). Dabei konnten Zusammenhänge zwischen strukturellen Ähnlichkeiten von Adjuvantien und deren Wirkmechanismen gefunden werden. Tween 20 und Tween 80 beispielsweise deckten ein sehr breites Spektrum an Mechanismen ab, zeigten dabei aber keinen herausragenden Wirkmechanismus einen bestimmten Prozess betreffend. Im Gegensatz dazu wiesen die nicht-ethoxylierten Span 20 und Span 80 hauptsächlich nur den oberflächenaktiven Mechanismus auf. Fast alle Adjuvantien, die in dieser Arbeit analysiert wurden, stellen komplexe polydisperse Mischungen dar, denen eine komplizierte Verteilung von EO-Gruppen und aliphatischen Ketten zugrunde liegt. Aus den Ergebnissen der vorliegenden Arbeit kann gemutmaßt werden, dass Adjuvantien mit einer eher breiten EO-Verteilung ein breiteres Anwendungsspektrum bieten können als Adjuvantien mit einer kleineren Verteilung. Es lässt sich vermuten, dass durch das Vorhandensein einer enormen Vielzahl einzelner Moleküle, die jeweils einen individuellen spezifischen physikalisch-chemischen Charakter aufweisen, ein großes Spektrum von Eigenschaften bezüglich der Sprayapplikation abgedeckt wird. KW - Adjuvans KW - Pflanzenschutzmittel KW - Adjuvant KW - Plant Protection KW - Kutikula KW - Plant Protection Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-138159 ER - TY - JOUR A1 - Asthana, Manish Kumar A1 - Brunhuber, Bettina A1 - Mühlberger, Andreas A1 - Reif, Andreas A1 - Schneider, Simone A1 - Herrmann, Martin J. T1 - Preventing the Return of Fear Using Reconsolidation Update Mechanisms Depends on the Met-Allele of the Brain Derived Neurotrophic Factor Val66Met Polymorphism JF - International Journal of Neuropsychopharmacology N2 - Background: Memory reconsolidation is the direct effect of memory reactivation followed by stabilization of newly synthesized proteins. It has been well proven that neural encoding of both newly and reactivated memories requires synaptic plasticity. Brain derived neurotrophic factor (BDNF) has been extensively investigated regarding its role in the formation of synaptic plasticity and in the alteration of fear memories. However, its role in fear reconsolidation is still unclear; hence, the current study has been designed to investigate the role of the BDNF val66met polymorphism (rs6265) in fear memory reconsolidation in humans. Methods: An auditory fear-conditioning paradigm was conducted, which comprised of three stages (acquisition, reactivation, and spontaneous recovery). One day after fear acquisition, the experimental group underwent reactivation of fear memory followed by the extinction training (reminder group), whereas the control group (non-reminder group) underwent only extinction training. On day 3, both groups were subjected to spontaneous recovery of earlier learned fearful memories. The treat-elicited defensive response due to conditioned threat was measured by assessing the skin conductance response to the conditioned stimulus. All participants were genotyped for rs6265. Results: The results indicate a diminishing effect of reminder on the persistence of fear memory only in the Met-allele carriers, suggesting a moderating effect of the BDNF polymorphism in fear memory reconsolidation. Conclusions: Our findings suggest a new role for BDNF gene variation in fear memory reconsolidation in humans. KW - BDNF KW - brain derived neurotrophic factor KW - fear conditioning KW - genetics memory KW - reconsolidation Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-166217 VL - 19 IS - 6 ER - TY - JOUR A1 - Audehm, P. A1 - Schmidt, M. A1 - Brück, S. A1 - Tietze, T. A1 - Gräfe, J. A1 - Macke, S. A1 - Schütz, G. A1 - Goering, E. T1 - Pinned orbital moments - A new contribution to magnetic anisotropy JF - Scientific Reports N2 - Reduced dimensionality and symmetry breaking at interfaces lead to unusual local magnetic configurations, such as glassy behavior, frustration or increased anisotropy. The interface between a ferromagnet and an antiferromagnet is such an example for enhanced symmetry breaking. Here we present detailed X-ray magnetic circular dichroism and X-ray resonant magnetic reflectometry investigations on the spectroscopic nature of uncompensated pinned magnetic moments in the antiferromagnetic layer of a typical exchange bias system. Unexpectedly, the pinned moments exhibit nearly pure orbital moment character. This strong orbital pinning mechanism has not been observed so far and is not discussed in literature regarding any theory for local magnetocrystalline anisotropy energies in magnetic systems. To verify this new phenomenon we investigated the effect at different temperatures. We provide a simple model discussing the observed pure orbital moments, based on rotatable spin magnetic moments and pinned orbital moments on the same atom. This unexpected observation leads to a concept for a new type of anisotropy energy. KW - pinned orbital moments KW - ferromagnet KW - antiferromagnet KW - anisotropy energy Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-167727 VL - 6 IS - 25517 ER - TY - JOUR A1 - Babski, Julia A1 - Haas, Karina A. A1 - Näther-Schindler, Daniela A1 - Pfeiffer, Friedhelm A1 - Förstner, Konrad U. A1 - Hammelmann, Matthias A1 - Hilker, Rolf A1 - Becker, Anke A1 - Sharma, Cynthia M. A1 - Marchfelder, Anita A1 - Soppa, Jörg T1 - Genome-wide identification of transcriptional start sites in the haloarchaeon Haloferax volcanii based on differential RNA-Seq (dRNA-Seq) JF - BMC Genomics N2 - Background Differential RNA-Seq (dRNA-Seq) is a recently developed method of performing primary transcriptome analyses that allows for the genome-wide mapping of transcriptional start sites (TSSs) and the identification of novel transcripts. Although the transcriptomes of diverse bacterial species have been characterized by dRNA-Seq, the transcriptome analysis of archaeal species is still rather limited. Therefore, we used dRNA-Seq to characterize the primary transcriptome of the model archaeon Haloferax volcanii. Results Three independent cultures of Hfx. volcanii grown under optimal conditions to the mid-exponential growth phase were used to determine the primary transcriptome and map the 5′-ends of the transcripts. In total, 4749 potential TSSs were detected. A position weight matrix (PWM) was derived for the promoter predictions, and the results showed that 64 % of the TSSs were preceded by stringent or relaxed basal promoters. Of the identified TSSs, 1851 belonged to protein-coding genes. Thus, fewer than half (46 %) of the 4040 protein-coding genes were expressed under optimal growth conditions. Seventy-two percent of all protein-coding transcripts were leaderless, which emphasized that this pathway is the major pathway for translation initiation in haloarchaea. A total of 2898 of the TSSs belonged to potential non-coding RNAs, which accounted for an unexpectedly high fraction (61 %) of all transcripts. Most of the non-coding TSSs had not been previously described (2792) and represented novel sequences (59 % of all TSSs). A large fraction of the potential novel non-coding transcripts were cis-antisense RNAs (1244 aTSSs). A strong negative correlation between the levels of antisense transcripts and cognate sense mRNAs was found, which suggested that the negative regulation of gene expression via antisense RNAs may play an important role in haloarchaea. The other types of novel non-coding transcripts corresponded to internal transcripts overlapping with mRNAs (1153 iTSSs) and intergenic small RNA (sRNA) candidates (395 TSSs). Conclusion This study provides a comprehensive map of the primary transcriptome of Hfx. volcanii grown under optimal conditions. Fewer than half of all protein-coding genes have been transcribed under these conditions. Unexpectedly, more than half of the detected TSSs belonged to several classes of non-coding RNAs. Thus, RNA-based regulation appears to play a more important role in haloarchaea than previously anticipated. KW - Archaea KW - dRNA-Seq KW - Promoter KW - Non-coding RNAs KW - sRNA KW - Haloferax volcanii KW - Transcriptome KW - Leaderless transcript KW - Antisense RNA Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-164553 VL - 17 IS - 629 ER - TY - THES A1 - Backhaus, Philipp T1 - Effects of Transgenic Expression of Botulinum Toxins in Drosophila T1 - Effekte der transgenen Expression von Botulinumtoxinen in Drosophila N2 - Clostridial neurotoxins (botulinum toxins and tetanus toxin) disrupt neurotransmitter release by cleaving neuronal SNARE proteins. We generated transgenic flies allowing for conditional expression of different botulinum toxins and evaluated their potential as tools for the analysis of synaptic and neuronal network function in Drosophila melanogaster by applying biochemical assays and behavioral analysis. On the biochemical level, cleavage assays in cultured Drosophila S2 cells were performed and the cleavage efficiency was assessed via western blot analysis. We found that each botulinum toxin cleaves its Drosophila SNARE substrate but with variable efficiency. To investigate the cleavage efficiency in vivo, we examined lethality, larval peristaltic movements and vision dependent motion behavior of adult Drosophila after tissue-specific conditional botulinum toxin expression. Our results show that botulinum toxin type B and botulinum toxin type C represent effective alternatives to established transgenic effectors, i.e. tetanus toxin, interfering with neuronal and non-neuronal cell function in Drosophila and constitute valuable tools for the analysis of synaptic and network function. N2 - Die verschiedenen Toxine der Bakterienspezies Clostridium (Botulinumtoxine und Tetanustoxin) interferieren mit Neuroexozytose durch Proteolyse der SNARE-Proteine. Wir haben transgene Fliegen generiert, die die Möglichkeit bieten konditional verschiedene Botulinumtoxine zu exprimieren. Durch biochemische Untersuchungen und Verhaltensexperimente haben wir das Potential dieser Toxine als Werkzeuge für die Analyse von Synapsen- und Netzwerkfunktion in Drosophila evaluiert. Durch Western Blot-Analysen stellten wir eine variierende Proteolysierbarkeit der Drosophila SNARE-Substrate durch die verschiedenen Botulinumtoxine dar. In Vivo untersuchten wir die Auswirkungen einer Zell-spezifischen Expression auf die Motorik in Larven und auf die Sehfähigkeit in adulten Fliegen. Unsere Resultate zeigen, dass Botulinumtoxin Typ B und C vielversprechende Alternativen zu etablierten molekularen Werkzeugen, wie Tetanustoxin, darstellen, um synaptische Transmission oder höhere Netzwerkfunktionen aufzuschlüsseln. Hierbei führt Botulinumtoxin Typ B zu einem spezifischen Verlust von neuronaler Aktivität, während Botulinumtoxin Typ C mit nicht Neuronen-spezifischer Zellfunktion interferiert. KW - Botulinustoxin KW - Drosophila KW - Synaptische Transmission KW - Behavioral neuroscience KW - molecular neuroscience KW - neurotoxins KW - SNARE proteins KW - neurotransmission Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-143279 ER - TY - THES A1 - Bahník, Štěpán T1 - Processing fluency and judgment T1 - Verarbeitungsflüssigkeit und Urteilsbildung N2 - To simplify a judgment, people often base it on easily accessible information. One cue that is usually readily available is processing fluency – a metacognitive feeling of ease of cognitive processing. Consequently, processing fluency is used as a cue for many different types of judgment, such as judgment of truth, confidence, and novelty. The present work describes results of three studies investigating various aspects of processing fluency effects on judgment. Processing fluency has been sometimes equated with speed of a cognitive process. Therefore, response times have been used for evaluation of processing fluency. However, response times in experimental tasks often do not encompass only the time needed for a given process, but also the time needed for a decision based on the resulting information. The study described in Chapter II uses a novel experimental method that enables separation of reading and decision times. The results show that people make a decision about liking of pseudowords faster when the pseudowords are hard-to-pronounce (i.e., disfluent) than when they are moderate in pronounceability. This suggests that response times cannot be used as a proxy for processing fluency when they include the time needed to make a decision. One of the studies of judgmental effects of processing fluency showed that food additives with easier pronounceable names are judged to be less harmful than those with hard-to-pronounce names. While people encounter food additives that are safe more often, this environmental association may be in the opposite direction for some categories of objects. For example, people are more likely to see names of especially dangerous criminals in the news. Chapter III describes a study which initially tested whether the fluency-safety association may be in the opposite direction for some categories of objects as a consequence of this selective exposure to especially dangerous exemplars. The results did not show support for this hypothesis. Furthermore, subsequent studies suggest that the previously found association between fluency and safety is replicable with the original stimuli used in the previous research, but not with newly constructed stimuli. Chapter IV describes a study which applied a finding from the processing fluency literature to a positive psychology exercise in order to increase its effectiveness. Namely, the experiment manipulated the number of good things that participants listed daily for two weeks as part of the exercise. While listing more things was considered harder, the number of things listed each day had no effect on effectiveness of the exercise. N2 - Um Urteilsprozesse zu vereinfachen, basieren Menschen diese oft auf leicht zugänglichen Informationen. Eine hierfür immer und jederzeit verfügbare Information ist die Verarbeitungsflüssigkeit – ein metakognitives Gefühl von Leichtigkeit bei der kognitiven Verarbeitung. Verarbeitungsflüssigkeit wird als Information bei vielen verschiedenen Arten von Urteilen genutzt, so beispielsweise bei Urteilen hinsichtlich Wahrheit, Vertrauenswürdigkeit und Neuheit. Die vorliegende Arbeit beschreibt die Ergebnisse von drei Studien, die verschiedene Aspekte des Einflusses von Verarbeitungsflüssigkeit auf Urteilsprozesse untersuchen. Verarbeitungsflüssigkeit wurde in der Literatur manchmal mit der Geschwindigkeit eines kognitiven Prozesses gleichgesetzt. Daher wurden Reaktionszeiten zur Schätzung der Verarbeitungsflüssigkeit herangezogen. In experimentellen Paradigmen umfassen Reaktionszeiten allerdings oft nicht nur die Zeit, die für den Prozess von Interesse benötigt wird, sondern auch die Zeit, die für eine Entscheidung benötigt wird, die auf der aus dem Prozess resultierenden Information basiert. Die Studie, die in Kapitel II beschrieben wird, verwendet daher eine neue experimentelle Methode, die eine Differenzierung zwischen der Zeit, die zum Lesen, und der Zeit, die für die Entscheidung benötigt wurde, ermöglicht. In den Ergebnissen zeigt sich, dass Menschen schneller eine Entscheidung darüber fällen, wie sehr sie bestimmte Pseudowörter mögen, wenn die Pseudowörter schwer aussprechbar sind (also nicht flüssig verarbeitbar sind) als wenn sie moderat aussprechbar sind. Dieser Befund legt nahe, dass Reaktionszeiten nicht zur Schätzung von Verarbeitungsflüssigkeit herangezogen werden können, da sie auch die Zeit beinhalten, die gebraucht wird, um eine Entscheidung zu treffen. Eine Studie zum Einfluss von Verarbeitungsflüssigkeit auf Urteilsbildung zeigte, dass Lebensmittelzusatzstoffe mit leicht aussprechbaren Namen im Vergleich zu solchen mit schwer aussprechbaren Namen als weniger gefährlich beurteilt wurden. Während Menschen ungefährlichen Lebensmittelzusatzstoffen öfter begegnen als gefährlichen, könnte dieser Umweltzusammenhang bei anderen Objektkategorien in entgegengesetzter Richtung verlaufen. Zum Beispiel sehen Menschen die Namen von besonders gefährlichen Verbrechern mit einer höheren Wahrscheinlichkeit in den Nachrichten. In Kapitel III wird daher eine Studie beschrieben, die ursprünglich testen sollte, ob der Verarbeitungsflüssigkeit-Sicherheit-Zusammenhang bei manchen Objektkategorien, bei denen man selektiv besonders gefährlichen Exemplaren ausgesetzt ist, in die entgegengesetzte Richtung verläuft. Die Ergebnisse unterstützten diese Hypothese jedoch nicht. Zudem lassen die Ergebnisse weiterer Studien darauf schließen, dass der zuvor gezeigte Zusammenhang zwischen Verarbeitungsflüssigkeit und Sicherheit nur mit den Originalstimuli aus früheren Studien, nicht aber mit neu konstruierten Studien repliziert werden kann. In Kapitel IV wird eine Studie beschrieben, die einen Befund der Verarbeitungsflüssigkeitsliteratur auf eine Übung aus dem Bereich der Positiven Psychologie anwendet, um deren Effektivität zu steigern. Konkret manipulierte das Experiment die Anzahl an guten Erlebnissen, die die Teilnehmer zwei Wochen lang täglich als Teil der Übung auflisten sollten. Obwohl das Auflisten einer größeren Anzahl an Erlebnissen als schwieriger bewertet wurde, hatte die Anzahl an Erlebnissen, die jeden Tag aufgelistet wurde, keinen Einfluss auf die Effektivität der Übung. KW - Urteilen KW - Judgment KW - Processing fluency KW - Informationsverarbeitung KW - Psychologie Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-144656 ER - TY - JOUR A1 - Bahník, Štěpán A1 - Strack, Fritz T1 - Overlap of accessible information undermines the anchoring effect JF - Judgment and Decision Making N2 - According to the Selective Accessibility Model of anchoring, the comparison question in the standard anchoring paradigm activates information that is congruent with an anchor. As a consequence, this information will be more likely to become the basis for the absolute judgment which will therefore be assimilated toward the anchor. However, if the activated information overlaps with information that is elicited by the absolute judgment itself, the preceding comparative judgment should not exert an incremental effect and should fail to result in an anchoring effect. The present studies find this result when the comparative judgment refers to a general category and the absolute judgment refers to a subset of the general category that was activated by the anchor value. For example, participants comparing the average annual temperature in New York City to a high 102 °F judged the average winter, but not summer temperature to be higher than participants making no comparison. On the other hand, participants comparing the annual temperature to a low –4 °F judged the average summer, but not winter temperature to be lower than control participants. This pattern of results was shown also in another content domain. It is consistent with the Selective Accessibility Model but difficult to reconcile with other main explanations of the anchoring effect. KW - anchoring KW - judgment KW - heuristics and biases KW - selective accessibility Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-169287 VL - 11 IS - 1 ER - TY - JOUR A1 - Baier, Pablo A. A1 - Baier-Saip, Jürgen A. A1 - Schilling, Klaus A1 - Oliveira, Jauvane C. T1 - Simulator for Minimally Invasive Vascular Interventions: Hardware and Software JF - Presence N2 - In the present work, a simulation system is proposed that can be used as an educational tool by physicians in training basic skills of minimally invasive vascular interventions. In order to accomplish this objective, initially the physical model of the wire proposed by Konings has been improved. As a result, a simpler and more stable method was obtained to calculate the equilibrium configuration of the wire. In addition, a geometrical method is developed to perform relaxations. It is particularly useful when the wire is hindered in the physical method because of the boundary conditions. Then a recipe is given to merge the physical and the geometrical methods, resulting in efficient relaxations. Moreover, tests have shown that the shape of the virtual wire agrees with the experiment. The proposed algorithm allows real-time executions, and furthermore, the hardware to assemble the simulator has a low cost. KW - simulation system KW - educational tool KW - invasive vascular interventions Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-140580 SN - 1531-3263 VL - 25 IS - 2 ER - TY - JOUR A1 - Banales, Jesus M. A1 - Cardinale, Vincenzo A1 - Carpino, Guido A1 - Marzioni, Marco A1 - Andersen, Jesper B. A1 - Invernizzi, Pietro A1 - Lind, Guro E. A1 - Folseraas, Trine A1 - Forbes, Stuart J. A1 - Fouassier, Laura A1 - Geier, Andreas A1 - Calvisi, Diego F. A1 - Mertens, Joachim C. A1 - Trauner, Michael A1 - Benedetti, Antonio A1 - Maroni, Luca A1 - Vaquero, Javier A1 - Macias, Rocio I. R. A1 - Raggi, Chiara A1 - Perugorria, Maria J. A1 - Gaudio, Eugenio A1 - Boberg, Kirsten M. A1 - Marin, Jose J. G. A1 - Alvaro, Domenico T1 - Cholangiocarcinoma: current knowledge and future perspectives consensus statement from the European Network for the Study of Cholangiocarcinoma (ENS-CCA) JF - Nature Reviews Gastroenterology & Hepatology N2 - Cholangiocarcinoma (CCA) is a heterogeneous group of malignancies with features of biliary tract differentiation. CCA is the second most common primary liver tumour and the incidence is increasing worldwide. CCA has high mortality owing to its aggressiveness, late diagnosis and refractory nature. In May 2015, the "European Network for the Study of Cholangiocarcinoma" (ENS-CCA: www.enscca.org or www.cholangiocarcinoma.eu) was created to promote and boost international research collaboration on the study of CCA at basic, translational and clinical level. In this Consensus Statement, we aim to provide valuable information on classifications, pathological features, risk factors, cells of origin, genetic and epigenetic modifications and current therapies available for this cancer. Moreover, future directions on basic and clinical investigations and plans for the ENS-CCA are highlighted. KW - primary sclerosing cholangitis KW - growth-factor-receptor KW - biliary-tract cancer KW - epithelial-mesenchymal transition KW - fine-needle-aspiration KW - human intrahepatic cholangiocarcinoma KW - induce cyclooxygenase-2 expression KW - human cholangiocellular carcinoma KW - nucleoside transporter KW - hepatic stellate cells Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-189077 VL - 13 IS - 5 ER - TY - JOUR A1 - Bankoglu, Ezgi Eyluel A1 - Tschopp, Oliver A1 - Schmitt, Johannes A1 - Burkard, Philipp A1 - Jahn, Daniel A1 - Geier, Andreas A1 - Stopper, Helga T1 - Role of PTEN in Oxidative Stress and DNA Damage in the Liver of Whole-Body Pten Haplodeficient Mice JF - PLoS One N2 - Type 2 diabetes (T2DM) and obesity are frequently associated with non-alcoholic fatty liver disease (NAFLD) and with an elevated cancer incidence. The molecular mechanisms of carcinogenesis in this context are only partially understood. High blood insulin levels are typical in early T2DM and excessive insulin can cause elevated reactive oxygen species (ROS) production and genomic instability. ROS are important for various cellular functions in signaling and host defense. However, elevated ROS formation is thought to be involved in cancer induction. In the molecular events from insulin receptor binding to genomic damage, some signaling steps have been identified, pointing at the PI3K/AKT pathway. For further elucidation Phosphatase and Tensin homolog (Pten), a tumour suppressor phosphatase that plays a role in insulin signaling by negative regulation of PI3K/AKT and its downstream targets, was investigated here. Dihydroethidium (DHE) staining was used to detect ROS formation in immortalized human hepatocytes. Comet assay and micronucleus test were performed to investigate genomic damage in vitro. In liver samples, DHE staining and western blot detection of HSP70 and HO-1 were performed to evaluate oxidative stress response. DNA double strand breaks (DSBs) were detected by immunohistostaining. Inhibition of PTEN with the pharmacologic inhibitor VO-OHpic resulted in increased ROS production and genomic damage in a liver cell line. Knockdown of Pten in a mouse model yielded increased oxidative stress levels, detected by ROS levels and expression of the two stress-proteins HSP70 and HO-1 and elevated genomic damage in the liver, which was significant in mice fed with a high fat diet. We conclude that PTEN is involved in oxidative stress and genomic damage induction in vitro and that this may also explain the in vivo observations. This further supports the hypothesis that the PI3K/AKT pathway is responsible for damaging effects of high levels of insulin. KW - insulin KW - mouse models DNA damage KW - oxidative stress KW - mammalian genomics KW - fatty liver KW - micronuclei KW - insulin signaling Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-146970 VL - 11 IS - 11 ER - TY - THES A1 - Bankoglu, Ezgi Eylül T1 - Oxidative status and genomic damage in an obesity model T1 - Oxidativer Status und Genom-Schäden in einem Adipositas-Modell N2 - Several cohort studies showed that obesity increases the risk of chronic disease such as T2DM, hypertension and non-alcoholic fatty liver disease and various types of cancer. Different factors were described that might be involving in these diseases in obesity. Some of these suggested factors were chronic infection, elevated free fatty acids, increased ROS formation, mitochondrial dysfunction and raised NAPDH oxidase activity. Obesity is a multifactorial disease and it is very hard to distinguish between all of these factors. In this study, we wanted to focus on the association between obesity, oxidative stress and genomic damage in kidney, liver and colon, which are the most relevant organs for cancer risk according to the cohort studies. Our findings indicated elevated oxidative stress in kidney, liver and colon together with elevated lipid, RNA and DNA oxidation in the whole body. Additionally, we were able to show increased DNA damage in kidney, liver and colon. Since obesity has become an epidemic all over the world, possible therapeutic applications such as life style changes (diet and sport), pharmacological supplements and various type of surgeries are increasing. As a second question, we focused on the effect of weight loss, which is supplied either by Roux-en-Y gastric bypass surgery or by caloric restriction designed in a way to provide the same extent of weight loss, on oxidative stress and genomic damage. Our results indicated that weight loss either by gastric bypass surgery or by caloric restriction led to reduced oxidative stress and genomic damage in kidney, liver and colon. We could not find any difference between the weight loss methods, except the DNA oxidation and repair marker urinary 8-oxodG, which was still elevated after RYGB, but not after caloric restriction. It is known that hyperinsulinemia and in the long term T2DM are among the biggest concerns in obese individuals. Since we know the mutagenic potential of elevated insulin levels from previous data in our working group, the correlation between the highly mutagenic DNA DBSs marker, γ-H2AX and the plasma insulin level was tested and the findings indicated a positive correlation. In order to demonstrate the association between insulin-related oxidative stress and genomic damage, we used in vitro and in vivo models with Pten deficiency. In this part of study, the work was focused on liver. Pten is a known negative regulator of the PI3K/Akt pathway, which is responsible for the elevated NADPH oxidase activity and mitochondrial dysfunction through elevated insulin levels. Pten inhibition or deficiency were used to sensitize the system to insulin. Non-transformed immortalized human hepatocytes were used to show the mutagenic potential of elevated insulin and these in vitro data revealed once more the link between insulin signaling, elevated oxidative stress and genomic damage. Since the metabolic function of the liver is not only due to the extent of the hepatic insulin response but is also affected by systemic interactions, a whole-body Pten haplodeficient mouse model with an additional Pten+/-/Akt2-/- group was utilized for in vivo investigation of insulin-mediated toxicity. Our findings in this model suggested that Pten deficiency alone can cause an increase in oxidative stress. HFD alone was sufficient to increase the expression of HO-1 and genomic damage significantly. Moreover, the combination (whole-body Pten haplodeficient mice fed with HFD) showed significantly elevated oxidative stress and genomic damage in mouse liver. However, Akt2 knockout could only reduce the oxidative stress and DNA damage in high fat diet fed mice significantly. All these findings demonstrated that obesity can induce oxidative stress and genomic damage. Elevated insulin levels are associated with obesity-mediated oxidative stress and genomic damage. However, the underlying mechanisms are surely multifaceted and complicated. For example, Pten as oncogene might also induce other mechanisms besides the elevation of the PI3K/Akt pathway activity. In conclusion, it is clear that oxidative stress and DNA damage are linked to obesity and that weight loss can reduce these two factors. Since DNA-damage is associated with an elevated cancer risk, it might be logical to use an antioxidant therapy in obese individuals to reduce the side effects and oxidative stress dependent mutagenicity and cancer risk in these individuals. However, much more research will be needed to support this idea experimentally. N2 - Mehrere Kohorten-studien zeigten, dass Adipositas das Risiko chronischer Erkrankungen wie Diabetes Mellitus Typ 2 (T2DM), Bluthochdruck, nicht-alkoholische Fettleber sowie das Risiko für unterschiedliche Krebsarten erhöht. Verschiedene Faktoren, die in Zusammenhang mit den Erkrankungen stehen, die Adipositas verursachen wurden bereits beschrieben. Einige dieser möglichen Faktoren sind chronische Infektionen, gesteigerte freie Fettsäuren, sowie reaktive Sauerstoffradikale, mitochondriale Dysfunktion und erhöhte Aktivität von NADPH-Oxidase. Adipositas ist eine multifaktorielle Erkrankung und unter von diesen Faktoren schwierig zu trennen. In dieser Studie wurde der Schwerpunkt auf den Zusammenhang von Adipositas, oxidativem Stress und Genomschäden in der Niere, Leber und dem Darm gelegt. Diese Organe sind gemäß der Kohortenstudien die anfälligsten hinsichtlich des Krebsrisikos. Unsere Befunde zeigten einen erhöhten oxidativen Stress in Niere, Leber und Darm, zusammen mit gesteigerter systemischer RNA-, DNA- und Fettoxidation, detektierbar anhand von Urinmarkern. Zusätzlich konnte eine Zunahme von DNA-Schäden in Niere, Leber und Darm aufgezeigt werden. Da Adipositas weltweit eine Epidemie geworden ist, nehmen mögliche therapeutische Anwendungen sowie eine Änderung des Lebensstils (Diät und Sport), pharmazeutische Ergänzungsmittel und verschiedene Arten von chirurgischen Behandlungen zu. Hier wurde der Fokus auf die Wirkung des Gewichtsverlustes, der durch Roux-en-Y Magen-Bypass-Chirurgie oder durch Kalorienreduzierung mit der Vorgabe eines gleichen Ausmaßes an Gewichtsverlust vorgegeben war, auf die Intensität des oxidativen Stress und des Genomschadens gerichtet. Unsere Befunde zeigten, dass der Gewichtsverlust sowohl durch Magen-Bypass-Chirurgie als auch durch Kalorienreduzierung zu einem reduzierten oxidativem Stress und Genomschaden in der Niere, der Leber und im Darm führten. Es konnte kein Unterschied zwischen den Methoden zur Reduzierung des Gewichtes gefunden werden, außer bei der DNA-Oxidation und dem Reparaturmarker 8-oxodG im Urin, der nach der RYGB immer noch erhöht war, aber nicht nach der Kalorienreduzierung. Es ist bekannt, dass Hyperinsulinämie bzw. Diabetes Mellitus Typ 2 eines der häufigsten Probleme bei übergewichtigen Patienten ist. Da wir das mutagene Potenzial von erhöhten Insulinspiegeln aus vorherigen Daten unserer Arbeitsgruppe kannten, wurde der Zusammenhang zwischen dem hoch mutagenen DNA-DSBs-Marker γ-H2AX und dem Plasma-Insulinspiegel analysiert. Die Befunde wiesen eine positive Korrelation auf. Um die Beziehung zwischen Insulin-verursachtem oxidativem Stress und Genomschaden aufzuzeigen, wurden in-vitro und in-vivo-Modelle mit Pten-Mangel benutzt. In diesem Teil der Studie wurde das Augenmerk auf die Leber gelegt. Das Protein Pten ist als negativer Regulator des PI3K/Akt Signalwegs bekannt, der unter anderem für die erhöhte Aktivität von NADPH Oxidase und mitochondrielle Dysfunktion durch erhöhten Insulinspiegel verantwortlich ist. Pten-Hemmung oder Pten-Mangel wurde genutzt, um unsere Versuchsmodelle für Insulin zu sensibilisieren. Nicht transformierbare immortalisierte menschliche Hepatozyten wurden verwendet, um das mutagene Potenzial von erhöhtem Insulin zu untersuchen, und die damit erzielten in -vitro-Daten wiesen wiederum auf die Beziehung zwischen Insulin-Signalwegen, oxidativem Stress und Genomschaden hin. Da die metabolische Funktion der Leber nicht nur dem Ausmaß der hepatischen Insulin-Reaktion geschuldet ist, sondern auch von systemischen Interaktionen beeinflusst wird, wurde ein Mausmodell für eine in-vivo-Untersuchung eingesetzt, das neben einem haploiden Pten-Mangel (Pten+/-) in einer Tiergruppe mit einer zusätzlichen Akt2-/- Defizienz (Pten+/-/Akt2-/-). Defizienz ausgestattet war. Unsere Befunde in diesem Modell zeigten, der Pten-Mangel alleine bereits erhöhten oxidativen-Stress verursachen kann. HFD war ebenfalls alleine bereits ausreichend, um die Expression von HO-1 und Genomschäden signifikant zu steigern. Darüber hinaus zeigte die Kombination (Pten-Mangel gefüttert mit HFD) eine signifikante Erhöhung des oxidativen Stresses und der Genomschäden in der Mäuseleber. Allerdings konnte das Fehlen von Akt2 den oxidativen Stress und Genomschaden nur in den mit HFD gefütterte Tieren signifikant verringern. Alle diese Befunde wiesen darauf hin, dass Adipositas oxidativen Stress und Genomschaden hervorrufen kann. Erhöhte Insulinspiegel sind mit Insulin-verursachtem oxidativem Stress und Genomschaden assoziiert. Allerdings sind die zugrunde liegenden Mechanismen sicherlich vielfältig und kompliziert. Zum Beispiel könnte Pten als Onkogen auch andere Mechanismen außer dem Anstieg der Aktivität des PI3K/Akt-Signalwegs- herbeiführen. Zusammenfassend ist es klar, dass oxidativer Stress und DNA-Schäden mit Adipositas zusammenhängen, und dass Gewichtsreduzierung diese zwei Faktoren verringern kann. Da DNA-Schäden mit erhöhtem Krebsrisiko assoziiert sind, könnte es folglich eine logische Konsequenz sein, Antioxidantien therapeutisch bei adipösen Patienten anzuwenden, um die Nebenwirkungen und die auf oxidativem Stress beruhende Mutagenität und das Krebsrisiko dieser Patienten zu verringern. Allerdings wird weitere intensive Forschung nötig sein, um dies mit experimentellen Daten zu untermauern.   KW - Übergewicht KW - DNS-Schädigung KW - Oxidativer Stress KW - DNA damage KW - Oxidative stress KW - Obesity KW - RYGB Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-137566 ER - TY - JOUR A1 - Baptista, Marisa A.P. A1 - Keszei, Marton A1 - Oliveira, Mariana A1 - Sunahara, Karen K.S. A1 - Andersson, John A1 - Dahlberg, Carin I.M. A1 - Worth, Austen J. A1 - Liedén, Agne A1 - Kuo, I-Chun A1 - Wallin, Robert P.A. A1 - Snapper, Scott B. A1 - Eidsmo, Liv A1 - Scheynius, Annika A1 - Karlsson, Mikael C.I. A1 - Bouma, Gerben A1 - Burns, Siobhan O. A1 - Forsell, Mattias N.E. A1 - Thrasher, Adrian J. A1 - Nylén, Susanne A1 - Westerberg, Lisa S. T1 - Deletion of Wiskott-Aldrich syndrome protein triggers Rac2 activity and increased cross-presentation by dendritic cells JF - Nature Communications N2 - Wiskott–Aldrich syndrome (WAS) is caused by loss-of-function mutations in theWASp gene. Decreased cellular responses in WASp-deficient cells have been interpreted to mean that WASp directly regulates these responses in WASp-sufficient cells. Here, we identify an exception to this concept and show that WASp-deficient dendritic cells have increased activation of Rac2 that support cross-presentation to CD8þ T cells. Using two different skin pathology models, WASp-deficient mice show an accumulation of dendritic cells in the skin and increased expansion of IFNg-producing CD8þ T cells in the draining lymph node and spleen. Specific deletion of WASp in dendritic cells leads to marked expansion of CD8þ T cells at the expense of CD4þ T cells. WASp-deficient dendritic cells induce increased cross-presentation to CD8þ T cells by activating Rac2 that maintains a near neutral pH of phagosomes. Our data reveals an intricate balance between activation of WASp and Rac2 signalling pathways in dendritic cells. KW - Cell signalling KW - Dendritic cells KW - Disease genetics Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-165966 VL - 7 ER - TY - JOUR A1 - Barbieri, Flavia L. A1 - Gardon, Jacques A1 - Ruiz-Castell, María A1 - Paco V., Pamela A1 - Muckelbauer, Rebecca A1 - Casiot, Corinne A1 - Freydier, Rémi A1 - Duprey, Jean-Louis A1 - Chen, Chih-Mei A1 - Müller-Nordhorn, Jacqueline A1 - Keil, Thomas T1 - Toxic trace elements in maternal and cord blood and social determinants in a Bolivian mining city JF - International Journal of Environmental Health Research N2 - This study assessed lead, arsenic, and antimony in maternal and cord blood, and associations between maternal concentrations and social determinants in the Bolivian mining city of Oruro using the baseline assessment of the ToxBol/Mine-Niño birth cohort. We recruited 467 pregnant women, collecting venous blood and sociodemographic information as well as placental cord blood at birth. Metallic/semimetallic trace elements were measured using inductively coupled plasma mass spectrometry. Lead medians in maternal and cord blood were significantly correlated (Spearman coefficient = 0.59; p < 0.001; 19.35 and 13.50 μg/L, respectively). Arsenic concentrations were above detection limit (3.30 μg/L) in 17.9 % of maternal and 34.6 % of cord blood samples. They were not associated (Fischer’s p = 0.72). Antimony medians in maternal and cord blood were weakly correlated (Spearman coefficient = 0.15; p < 0.03; 9.00 and 8.62 μg/L, respectively). Higher concentrations of toxic elements in maternal blood were associated with maternal smoking, low educational level, and partner involved in mining. KW - environmental exposure KW - metallic trace elements KW - maternal exposure KW - prenatal exposure KW - risk factors Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-150385 VL - 26 IS - 2 ER - TY - JOUR A1 - Barbieri, Flavia L. A1 - Gardon, Jacques A1 - Ruiz-Castell, María A1 - Paco V., Pamela A1 - Muckelbauer, Rebecca A1 - Casiot, Corinne A1 - Freydier, Rémi A1 - Duprey, Jean-Louis A1 - Chen, Chih-Mei A1 - Müller-Nordhorn, Jacqueline A1 - Keil, Thomas T1 - Toxic trace elements in maternal and cord blood and social determinants in a Bolivian mining city JF - International Journal of Environmental Health Research N2 - This study assessed lead, arsenic, and antimony in maternal and cord blood, and associations between maternal concentrations and social determinants in the Bolivian mining city of Oruro using the baseline assessment of the ToxBol/Mine-Nino birth cohort. We recruited 467 pregnant women, collecting venous blood and sociodemographic information as well as placental cord blood at birth. Metallic/semimetallic trace elements were measured using inductively coupled plasma mass spectrometry. Lead medians in maternal and cord blood were significantly correlated (Spearman coefficient=0.59; p<0.001; 19.35 and 13.50 μg/L, respectively). Arsenic concentrations were above detection limit (3.30 μg/L) in 17.9% of maternal and 34.6% of cord blood samples. They were not associated (Fischer's p=0.72). Antimony medians in maternal and cord blood were weakly correlated (Spearman coefficient=0.15; p<0.03; 9.00 and 8.62 μg/L, respectively). Higher concentrations of toxic elements in maternal blood were associated with maternal smoking, low educational level, and partner involved in mining. KW - environmental exposure KW - metallic trace elements KW - maternal exposure KW - prenatal exposure KW - risk factors Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-190703 VL - 26 IS - 2 ER - TY - JOUR A1 - Bargul, Joel L. A1 - Jung, Jamin A1 - McOdimba, Francis A. A1 - Omogo, Collins O. A1 - Adung'a, Vincent O. A1 - Krüger, Timothy A1 - Masiga, Daniel K. A1 - Engstler, Markus T1 - Species-Specific Adaptations of Trypanosome Morphology and Motility to the Mammalian Host JF - PLoS Pathogens N2 - African trypanosomes thrive in the bloodstream and tissue spaces of a wide range of mammalian hosts. Infections of cattle cause an enormous socio-economic burden in sub-Saharan Africa. A hallmark of the trypanosome lifestyle is the flagellate’s incessant motion. This work details the cell motility behavior of the four livestock-parasites Trypanosoma vivax, T. brucei, T. evansi and T. congolense. The trypanosomes feature distinct swimming patterns, speeds and flagellar wave frequencies, although the basic mechanism of flagellar propulsion is conserved, as is shown by extended single flagellar beat analyses. Three-dimensional analyses of the trypanosomes expose a high degree of dynamic pleomorphism, typified by the ‘cellular waveform’. This is a product of the flagellar oscillation, the chirality of the flagellum attachment and the stiffness of the trypanosome cell body. The waveforms are characteristic for each trypanosome species and are influenced by changes of the microenvironment, such as differences in viscosity and the presence of confining obstacles. The distinct cellular waveforms may be reflective of the actual anatomical niches the parasites populate within their mammalian host. T. vivax displays waveforms optimally aligned to the topology of the bloodstream, while the two subspecies T. brucei and T. evansi feature distinct cellular waveforms, both additionally adapted to motion in more confined environments such as tissue spaces. T. congolense reveals a small and stiff waveform, which makes these parasites weak swimmers and destined for cell adherence in low flow areas of the circulation. Thus, our experiments show that the differential dissemination and annidation of trypanosomes in their mammalian hosts may depend on the distinct swimming capabilities of the parasites. KW - swimming KW - viscosity KW - flagella KW - host-pathogen interactions KW - cell motility KW - blood KW - parasitic diseases KW - trypanosoma brucei gambiense Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-146513 VL - 12 IS - 2 ER - TY - JOUR A1 - Barquist, Lars A1 - Mayho, Matthew A1 - Cummins, Carla A1 - Cain, Amy K. A1 - Boinett, Christine J. A1 - Page, Andrew J. A1 - Langridge, Gemma C. A1 - Quail, Michael A. A1 - Keane, Jacqueline A. A1 - Parkhill, Julian T1 - The TraDIS toolkit: sequencing and analysis for dense transposon mutant libraries JF - Bioinformatics N2 - Transposon insertion sequencing is a high-throughput technique for assaying large libraries of otherwise isogenic transposon mutants providing insight into gene essentiality, gene function and genetic interactions. We previously developed the Transposon Directed Insertion Sequencing (TraDIS) protocol for this purpose, which utilizes shearing of genomic DNA followed by specific PCR amplification of transposon-containing fragments and Illumina sequencing. Here we describe an optimized high-yield library preparation and sequencing protocol for TraDIS experiments and a novel software pipeline for analysis of the resulting data. The Bio-Tradis analysis pipeline is implemented as an extensible Perl library which can either be used as is, or as a basis for the development of more advanced analysis tools. This article can serve as a general reference for the application of the TraDIS methodology. KW - mechanisms KW - Transposon insertion sequencing KW - sequencing protocol KW - TraDIS Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-189667 VL - 32 IS - 7 ER - TY - JOUR A1 - Bartelheim, Kerstin A1 - Nemes, Karolina A1 - Seeringer, Angela A1 - Kerl, Kornelius A1 - Buechner, Jochen A1 - Boos, Joachim A1 - Graf, Norbert A1 - Dürken, Matthias A1 - Gerss, Joachim A1 - Hasselblatt, Martin A1 - Kortmann, Rolf-Dieter A1 - Teichert von Luettichau, Irene A1 - Nagel, Inga A1 - Nygaard, Randi A1 - Oyen, Florian A1 - Quiroga, Eduardo A1 - Schlegel, Paul-Gerhardt A1 - Schmid, Irene A1 - Schneppenheim, Reinhard A1 - Siebert, Reiner A1 - Solano-Paez, Palma A1 - Timmermann, Beate A1 - Warmuth-Metz, Monika A1 - Frühwald, Michael Christoph T1 - Improved 6-year overall survival in AT/RT - results of the registry study Rhabdoid 2007 JF - Cancer Medicine N2 - Atypical teratoid rhabdoid tumors (AT/RT) are characterized by mutations and subsequent inactivation of SMARCB1 (INI1, hSNF5), a predilection for very young children and an unfavorable outcome. The European Registry for rhabdoid tumors (EU‐RHAB) was established to generate a common European database and to establish a standardized treatment regimen as the basis for phase I/II trials. Thus, genetic analyses, neuropathologic and radiologic diagnoses, and a consensus treatment regimen were prospectively evaluated. From 2005 to 2009, 31 patients with AT/RT from four countries were recruited into the registry study Rhabdoid 2007 and treated with systemic and intraventricular chemotherapy. Eight patients received high‐dose chemotherapy, 23 radiotherapy, and 17 maintenance therapy. Reference evaluations were performed in 64% (genetic analyses, FISH, MLPA, sequencing) up to 97% (neuropathology, INI1 stain). Germ‐line mutations (GLM) were detected in 6/21 patients. Prolonged overall survival was associated with age above 3 years, radiotherapy and achievement of a complete remission. 6‐year overall and event‐free survival rates were 46% (±0.10) and 45% (±0.09), respectively. Serious adverse events and one treatment‐related death due to insufficiency of a ventriculo peritoneal shunt (VP‐shunt) and consecutive herniation were noted. Acquisition of standardized data including reference diagnosis and a standard treatment schedule improved data quality along with a survival benefit. Treatment was feasible with significant but manageable toxicity. Although our analysis is biased due to heterogeneous adherence to therapy, EU‐RHAB provides the best available basis for phase I/II clinical trials. KW - AT/RT KW - EU‐RHAB Registry KW - pediatric brain tumor KW - Rhabdoid 2007 Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-164799 VL - 5 IS - 8 ER - TY - INPR A1 - Bartfeld, Sina T1 - Modeling infectious diseases and host-microbe interactions in gastrointestinal organoids T2 - Developmental Biology N2 - Advances in stem cell research have allowed the development of 3-dimensional (3D) primary cell cultures termed organoid cultures, as they closely mimic the in vivo organization of different cell lineages. Bridging the gap between 2-dimensional (2D) monotypic cancer cell lines and whole organisms, organoids are now widely applied to model development and disease. Organoids hold immense promise for addressing novel questions in host-microbe interactions, infectious diseases and the resulting inflammatory conditions. Researchers have started to use organoids for modeling infection with pathogens, such as Helicobacter pylori or Salmonella enteritica, gut- microbiota interactions and inflammatory bowel disease. Future studies will broaden the spectrum of microbes used and continue to establish organoids as a standard model for human host-microbial interactions. Moreover, they will increasingly exploit the unique advantages of organoids, for example to address patient-specific responses to microbes. KW - gastrointestinal disease KW - salmonella KW - microbiota KW - inflammatory bowel disease KW - organoid culture KW - helicobacter KW - rotavirus KW - norovirus Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-138788 UR - http://www.sciencedirect.com/science/article/pii/S0012160616304602 SN - 0012-1606 N1 - This is the accepted version of the following article: Bartfeld, Sina, Modeling infectious diseases and host-microbe interactions in gastrointestinal organoids, Developmental Biology, 2016, 420, 2, 262-270. http://dx.doi.org/10.1016/j.ydbio.2016.09.014 ER - TY - THES A1 - Batsching, Sophie Johanna T1 - Behavior under uncontrollable stress in \(Drosophila\) \(melanogaster\) - Learned Helplessness revisited T1 - Verhalten unter nicht kontrollierbarem Stress - Neubetrachtung der Erlernten Hilflosigkeit bei \(Drosophila\) \(melanogaster\) N2 - In order to select the appropriate behavior, it is important to choose the right behavior at the right time out of many options. It still remains unclear nowadays how exactly this is managed. To address this question, I expose flies (Drosophila melanogaster) to uncontrollable stress to study their behavior under restrictive circumstances by using the so-called shock box. Exposing animals to uncontrollable stress may have an impact on subsequent behavior and can last for some time. The animal learns that whatever it does, it cannot change the situation and therefore can develop something called learned helplessness. The term was first conceptualized by two American psychologists Maier and Seligman (1967), who discovered this phenomenon while doing experiments with dogs. They found out that dogs which are exposed to inescapable stress, later fail in a learning task (‘shuttle box’). In this work the walking patterns of three different types of experimental flies, walking in a small dark chamber, were evaluated. Using the triadic design (Seligman and Maier, 1967), flies were either exposed to electric shock randomly (yoked), could turn it off by being active (master) or did not receive punishment at all (control). Master flies were shocked whenever they sat for more than 0.9 seconds. At the same time yoked flies received a shock as well independent of what they were doing, to ensure the same amount of shocks received and to create random punishment pattern for the yoked group. With this so-called no-idleness paradigm flies were conditioned either 10 minutes, which resulted in a short (3 minutes) after-effect, or 20 minutes that turned out to be more stable (10 minutes). In a second part, the behavior during the 20 minute conditioning and a 10 minutes post-test was described in detail. Female flies of the yoked group developed lower activity levels, longer pauses and walked more slowly than master and control flies during conditioning. In the time after the shocks while still in the box, the yoked flies also reduced the frequency and duration of walking bouts as well as their walking speed. Additionally, they took more time to resume walking after the onset of an electric shock than master flies (escape latency) and turned out to make less pauses lasting between 1-1.5 seconds which supports the finding concerning the escape latency. Male flies, tested under the same conditions, showed a slightly weaker after-effect regarding the difference between master and yoked during conditioning and post-test when compared to female flies. When comparing the 20 minutes conditioning with subsequent 10 minutes test in the heat and the shock box in parallel, one finds the same effect: Flies which do not have control over the shocks, lower their activity, make less but longer pauses and walk more slowly than their respective master flies. Despite the similar effect of heat and shock on the flies, some differences between the devices occurred, which can partly be explained by different humidity conditions as well as by different surfaces within the chambers. When the control over the shocks is given back to the yoked flies, it takes them about seven minutes to realize it. One could also show that dopamine levels in the brain were reduced in comparison to flies which did not receive shocks. Yoked flies also were impaired in a place learning task (place learning) and their reaction to light (exit from the box towards the light) directly after conditioning. After characterizing the walking behavior in the chambers, the study deals with the question whether the effects observed in the chambers transfer to different environments. In free walk they only differed from flies which did not receive electric shocks and no effect of uncontrollability was transferred to courtship behavior. Handling as the cause could be excluded. Since handling could be exclude to be the cause of losing the effect, I assumed that the behavior shown in the boxes are context depend. Not only were the after-effects of inescapable shock subject of the current research also the impact of the rearing situation on the response to electric shock was investigated in the present study. Flies which grew up in a single-reared situation turned out to be less affected by inescapable stress in both sexes. In the next part, the first steps to unravel the neuronal underpinning were taken. A mutant – fumin – which is defective in the dopamine re-uptake transporter showed less reaction to inescapable foot shocks, while a mutant for the gene which encodes an adenylate cyclase (rutabaga2080) resulted in a good score during conditioning, but showed no stable after-effect. Downregulating the expression of the adenylate cyclase gene (rutabaga) in different parts of the mushroom bodies showed, that rutabaga is necessary in the α’β’-lobes for expressing the differences between master and yoked flies in the no-idleness paradigm. The study further confirmed previous findings, that rutabaga is needed in operant but not in classical conditioning. As a result, the study could show that not the stimulus itself causes the state of uncontrollability but the fact that the fly learned that it was not in control of the stimulus. This state turned out to be context and time dependent. N2 - Eine wichtige Aufgabe für ein Tier ist es, das passende Verhalten zur richtigen Zeit zu wählen. Heutzutage ist immer noch unklar, wie dieser Prozess exakt abläuft. Zur Untersuchung dieser Frage werden Fliegen (Drosophila melanogaster) in der so genannten Schockbox unkontrollierbarem Stress ausgesetzt um auf diesem Weg Verhaltenskontrolle unter stressigen und stark restriktiven Umständen untersuchen zu können. Wenn Tiere unkontrollierbarem Stress ausgesetzt sind, kann dieser Zustand sowohl langanhaltend sein als auch Einfluss auf das Folgeverhalten haben. Das Tier lernt, dass alle Aktivitäten, die es in dieser Situation unternimmt keinen Einfluss auf die Situation haben. Dadurch kann das Tier einen Zustand der sogenannten Erlernten Hilflosigkeit entwickeln. Dieser Begriff wurde von zwei amerikanischen Psychologen, Maier und Seligman (1976), geprägt, die dieses Phänomen während Experimenten mit Hunden entdeckten und konzipierten. Sie fanden heraus, dass Hunde, die unkontrollierbarem Stress ausgesetzt waren, an einer anschließend gestellten Lernaufgabe scheiterten (‚shuttle-box‘). Gegenstand der vorliegenden Arbeit ist es, das Laufmuster in einer schmalen und kleinen Kammer an drei verschiedenen Versuchsgruppen von Fliegen zu analysieren. Unter Verwendung des sogenannten triadischen Konzepts (Seligman and Maier, 1967) wurden die Fliegen drei unterschiedlichen Situationen ausgesetzt: Zufällige Elektroschocks (Yoked-Gruppe), durch Laufen abschaltbare Elektroschocks (Master-Gruppe) oder keine Bestrafung (Kontroll-Gruppe). Master-Fliegen wurden immer dann geschockt, wenn sie für länger als 0,9 Sekunden saßen. Unabhängig ihres Verhaltens erhielten die Yoked-Fliegen zeitgleich einen Schock um einen zufälligen Bestrafungsreiz zu generieren. Mit diesem so genannten ‚no-idleness‘ (nicht ruhen dürfen) Paradigma wurden die Fliegen entweder zehn Minuten oder 20 Minuten konditioniert. Während eine zehnminütige Konditionierung zu einem kurzen Nacheffekt führte (Nacheffekt von drei Minuten), stellte sich die zwanzigminütige Konditionierung als nachhaltiger heraus (Nacheffekt von zehn Minuten). In einem zweiten Teil der Arbeit wurde das Verhalten der Fliegen sowohl während der zwanzig Minuten andauernden Konditionierung also auch im nachfolgenden zehnminütigen Test im Detail beschrieben. Während der zwanzigminütigen Konditionierung zeigten weibliche Yoked-Fliegen eine geringere Aktivität, saßen länger und liefen langsamer als Master- oder Kontroll-Fliegen. In der Zeit nach den Schocks, zeigten sie immer noch eine verminderte Lauffrequenz sowie kürzere und langsamere Laufphasen. Zusätzlich benötigten sie länger um nach dem Einsetzten eines Elektroschocks loszulaufen (Flucht-Latenzzeit) und machten weniger Kurzpausen die zwischen 1 bis 1,5 Sekunden lang waren. Dies unterstützt das Ergebnis der verlängerten Flucht-Latenzzeit. Männchen, die unter gleichen Bedingungen getestet wurden, wiesen im Vergleich zu weiblichen Fliegen eine leicht abgeschwächte Reaktion bezüglich des Master-Yoked-Unterschieds auf. Wenn die Konditionierung mit dem anschließenden Test in der Schock- und der Hitzekammer gleichzeitig durchgeführt wurde, resultierte dies in vergleichbaren Ergebnissen: Fliegen, die keine Kontrolle über den Reiz haben, vermindern ihr Aktivitätslevel, sitzen seltener aber länger und laufen langsamer als die dazugehörigen Master-Fliegen. Neben der Tatsache, dass ein ähnlicher Effekt auftritt, weisen die Apparaturen dennoch kleine Unterschiede auf. Diese können zu Teilen mit den unterschiedlichen Luftfeuchtigkeitsniveaus als auch durch die Verschiedenheit der Laufoberfläche der jeweiligen Kammern erklärt werden. Wird den Fliegen die Kontrolle über die Schocks zurückgegeben, benötigen sie etwa sieben Minuten um dies zu erkennen. Zudem konnte gezeigt werden, dass die Dopaminkonzentration in den Köpfen, im Vergleich zu Tieren die keine Schocks erhalten haben, vermindert war. Yoked-Fliegen wiesen außerdem unmittelbar nach der Konditionierung Defekte im Ortslernen und in ihrer positiven Reaktion auf Licht auf. Nachdem das Laufverhalten innerhalb der Kammern ausführlich charakterisiert wurde, geht diese Studie darauf ein, ob die Effekte, die in den Kammern gemessen wurden, auch in anderen Umgebungen zu beobachten sind. Im freien Lauf unterschieden sie sich lediglich von Fliegen, die keine Schocks erhalten hatten und es sind keine Auswirkungen durch Kontrollverlust im Paarungsverhalten festzustellen. Da die Handhabung der Tiere als Grund für den Verlust des Nacheffektes ausgeschlossen werden konnte, lässt sich schlussfolgern, dass das Verhalten das in den Kammern gemessen wurde, kontextabhängig ist. Zusätzlich zur Untersuchung der Auswirkungen unausweichlichen Stresses, wurde der Einfluss, der Aufzuchtbedingungen auf die Stress-Antwort in der vorliegenden Studie untersucht. Fliegen, die einzeln aufgezogen wurden, weisen bei beiden Geschlechtern eine verminderte Antwort auf Stress auf. Im darauffolgenden Abschnitt wurden erste Schritte unternommen, um die neuronalen Grundlagen der Erlernten Hilflosigkeit zu untersuchen. Eine Mutante – fumin – die ein defektes Wiederaufnahmetransporter-Gen für Dopamin besitzt, wies eine verminderte Stressantwort auf. Während eine Mutante des Adenylatzyklasegens (rutabaga2080) normale Ergebnisse während der Konditionierung aufzeigten, war im Post-test kein signifikanter Nacheffekt messbar. Das Herunterregulieren des Adenylatcyclasengens (rutabaga), in verschiedenen Teilen der Pilzkörper, zeigte dass die Expression von rutabaga in den α’β’-Loben für die Entwicklung der Erlernten Hilflosigkeit im no-idleness Paradigma benötigt wird. Zudem konnten vorangegangene Studien bestätigt werden, die rutabaga eine Rolle im operanten Lernen jedoch nicht im klassischen Lernen zuordnen. Als Fazit zeigt die Studie, dass nicht der Stressor selbst, sondern die Unkontrollierbarkeit des Stressors der Grund für die Entwicklung der Erlernten Hilflosigkeit darstellt und das Phänomen, innerhalb der hier gewählten Zeitspanne (20 Minuten Stress), kontextabhängig zu sein scheint. KW - Taufliege KW - Stress KW - Verhalten KW - Gelernte Hilflosigkeit KW - Erlernte Hilflosigkeit KW - Learned Helplessness KW - Behavior KW - Drosophila melanogaster Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-145416 ER - TY - JOUR A1 - Batsching, Sophie A1 - Wolf, Reinhard A1 - Heisenberg, Martin T1 - Inescapable Stress Changes Walking Behavior in Flies - Learned Helplessness Revisited JF - PLoS ONE N2 - Like other animals flies develop a state of learned helplessness in response to unescapable aversive events. To show this, two flies, one 'master', one 'yoked', are each confined to a dark, small chamber and exposed to the same sequence of mild electric shocks. Both receive these shocks when the master fly stops walking for more than a second. Behavior in the two animals is differently affected by the shocks. Yoked flies are transiently impaired in place learning and take longer than master flies to exit from the chamber towards light. After the treatment they walk more slowly and take fewer and shorter walking bouts. The low activity is attributed to the fly's experience that its escape response, an innate behavior to terminate the electric shocks, does not help anymore. Earlier studies using heat pulses instead of electric shocks had shown similar effects. This parallel supports the interpretation that it is the uncontrollability that induces the state. KW - learning KW - locomotion KW - animal behavior KW - behavioral conditioning KW - walking KW - vibration KW - light pulses KW - conditioned response Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-178640 VL - 11 IS - 11 ER - TY - JOUR A1 - Baumeister, Joachim A1 - Striffler, Albrecht A1 - Brandt, Marc A1 - Neumann, Michael T1 - Collaborative Decision Support and Documentation in Chemical Safety with KnowSEC JF - Journal of Cheminformatics N2 - To protect the health of human and environment, the European Union implemented the REACH regulation for chemical substances. REACH is an acronym for Registration, Evaluation, Authorization, and Restriction of Chemicals. Under REACH, the authorities have the task of assessing chemical substances, especially those that might pose a risk to human health or environment. The work under REACH is scientifically, technically and procedurally a complex and knowledge-intensive task that is jointly performed by the European Chemicals Agency and member state authorities in Europe. The assessment of substances under REACH conducted in the German Environment Agency is supported by the knowledge-based system KnowSEC, which is used for the screening, documentation, and decision support when working on chemical substances. The software KnowSEC integrates advanced semantic technologies and strong problem solving methods. It allows for the collaborative work on substances in the context of the European REACH regulation. We discuss the applied methods and process models and we report on experiences with the implementation and use of the system. KW - decision support KW - knowledge-based systems KW - ontologies KW - expert systems KW - semantic technologies Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-146575 VL - 8 IS - 21 ER - TY - JOUR A1 - Baur, Johannes A1 - Ritter, Christian O. A1 - Germer, Christoph-Thomas A1 - Klein, Ingo A1 - Kickuth, Ralph A1 - Steger, Ulrich T1 - Transarterial chemoembolization with drug-eluting beads versus conventional transarterial chemoembolization in locally advanced hepatocellular carcinoma JF - Hepatic Medicine N2 - Purpose: In hepatocellular carcinoma patients with large or multinodal tumors, where curative treatment options are not feasible, transarterial therapies play a major role. Transarterial chemoembolization (TACE) with drug-eluting beads (DEB-TACE) is a promising new approach due to higher intratumoral and lower systemic concentration of the chemotherapeutic agent compared to conventional TACE (cTACE). Patients and methods: In a retrospective analysis, 32 patients with hepatocellular carcinoma who received either DEB or a cTACE were compared regarding survival time, disease recurrence, and side effects such as pain and fever. Results: No significant differences could be detected between the cTACE and DEB-TACE groups with regard to mean hospital stay, appearance of postinterventional fever, or 30-day mortality. However, the application of intravenous analgesics as postinterventional pain medication was needed more often in patients treated with DEB-TACE (57.1% vs 12.5%, P=0.0281). The overall median survival after the initial procedure was 10.8 months in the cTACE group and 9.2 months in the DEB-TACE group, showing no significant difference. Conclusion: No survival benefit for patients treated with either DEB-TACE or cTACE was observed. Surprisingly, a higher rate of postinterventional pain could be detected after DEB-TACE. KW - transarterial chemoembolization KW - hepatocellular carcinoma KW - drug-eluting beads Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-146553 VL - 2016 IS - 8 ER - TY - THES A1 - Baur, Ramona T1 - Adult Attention-Deficit/Hyperactivity Disorder (ADHD), Emotion Processing, and Emotion Regulation in Virtual Reality T1 - Aufmerksamkeitsdefizit-/Hyperaktivitätsstörung (ADHS) im Erwachsenenalter, Emotionsverarbeitung und Emotionsregulation in virtueller Realität N2 - Attention-Deficit/Hyperactivity Disorder (ADHD) is characterized by symptoms of inattentiveness and hyperactivity/impulsivity. Besides, increasing evidence points to ADHD patients showing emotional dysfunctions and concomitant problems in social life. However, systematic research on emotional dysfunctions in ADHD is still rare, and to date most studies lack conceptual differentiation between emotion processing and emotion regulation. The aim of this thesis was to systematically investigate emotion processing and emotion regulation in adult ADHD in a virtual reality paradigm implementing social interaction. Emotional reactions were assessed on experiential, physiological, and behavioral levels. Experiment 1 was conducted to develop a virtual penalty kicking paradigm implying social feedback and to test it in a healthy sample. This paradigm should then be applied in ADHD patients later on. Pleasant and unpleasant trials in this paradigm consisted of hits respectively misses and subsequent feedback from a virtual coach. In neutral trials, participants were teleported to different spots of the virtual stadium. Results indicated increased positive affectivity (higher valence and arousal ratings, higher zygomaticus activations, and higher expression rates of positive emotional behavior) in response to pleasant compared to neutral trials. Reactions to unpleasant trials were contradictory, indicating increased levels of both positive and negative affectivity, compared to neutral trials. Unpleasant vs. neutral trials revealed lower valence ratings, higher arousal ratings, higher zygomaticus activations, slightly lower corrugator activations, and higher expression rates of both positive and negative emotional behavior. The intensity of emotional reactions correlated with experienced presence in the virtual reality. To better understand the impact of hits or misses per se vs. hits or misses with coach feedback healthy participants’ emotional reactions, only 50% of all shots were followed by coach feedback in experiment 2. Neutral trials consisted of shots over the free soccer field which were followed by coach feedback in 50 % of all trials. Shots and feedback evoked more extreme valence and arousal ratings, higher zygomaticus activations, lower corrugator activations, and higher skin conductance responses than shots alone across emotional conditions. Again, results speak for the induction of positive emotions in pleasant trials whereas the induction of negative emotions in unpleasant trials seems ambiguous. Technical improvements of the virtual reality were reflected in higher presence ratings than in experiment 1. Experiment 3 investigated emotional reactions of adult ADHD patients and healthy controls after emotion processing and response-focused emotion regulation. Participants successively went through an ostensible online ball-tossing game (cyber ball) inducing negative emotions, and an adapted version of the virtual penalty kicking game. Throughout cyber ball, participants were included or ostracized by two other players in different experimental blocks. Participants were instructed to explicitly show, not regulate, or hide their emotions in different experimental blocks. Results provided some evidence for deficient processing of positive emotions in ADHD. Patients reported slightly lower positive affect than controls during cyber ball, gave lower valence ratings than controls in response to pleasant penalty kicking trials, and showed lower zygomaticus activations than controls especially during penalty kicking. Patients in comparison with controls showed slightly increased processing of unpleasant events during cyber ball (higher ratings of negative affect, especially in response to ostracism), but not during penalty kicking. Patients showed lower baseline skin conductance levels than controls, and impaired skin conductance modulations. Compared to controls, patients showed slight over-expression of positive as well as negative emotional behavior. Emotion regulation analyses revealed no major difficulties of ADHD vs. controls in altering their emotional reactions through deliberate response modulation. Moreover, patients reported to habitually apply adaptive emotion regulation strategies even more frequently than controls. The analyses of genetic high-risk vs. low-risk groups for ADHD across the whole sample revealed similar results as analyses for patients vs. controls for zygomaticus modulations during emotion processing, and for modulations of emotional reactions due to emotion regulation. To sum up, the virtual penalty kicking paradigm proved to be successful for the induction of positive, but not negative emotions. The importance of presence in virtual reality for the intensity of induced emotions could be replicated. ADHD patients showed impaired processing of primarily positive emotions. Aberrations in negative emotional responding were less clear and need further investigation. Results point to adult ADHD in comparison to healthy controls suffering from baseline deficits in autonomic arousal and deficits in arousal modulation. Deficits of ADHD in the deliberate application of response-focused emotion regulation could not be found. N2 - Die Aufmerksamkeitsdefizit-/hyperaktivitätsstörung (ADHS) ist gekennzeichnet durch Symptome der Unaufmerksamkeit und Hyperaktivität/Impulsivität. Zudem sprechen zunehmende Befunde für emotionale Defizite und damit einhergehende soziale Probleme bei ADHS. Bisher gibt es jedoch kaum systematische Untersuchungen zu emotionalen Defiziten bei ADHS, und die meisten bisherigen Studien trennen nicht klar zwischen den Konzepten der Emotionsverarbeitung und Emotionsregulation. Das Ziel dieser Arbeit war es, Emotionsverarbeitung und Emotionsregulation bei erwachsenen ADHS Patienten in einem Paradigma in virtueller Realität mit sozialer Interaktion zu untersuchen. Emotionale Reaktionen wurden auf Erlebnisebene, physiologischer Ebene und Verhaltensebene erfasst. In Experiment 1 wurde ein virtuelles Elfmeterparadigma mit sozialem Feedback entwickelt und an einer gesunden Stichprobe getestet. Dieses Paradigma sollte später mit ADHS Patienten verwendet werden. Angenehme und unangenehme Versuchsdurchgänge bestanden aus Treffern bzw. gehaltenen Torschüssen und einer darauffolgenden Rückmeldung von einem virtuellen Trainer. In neutralen Durchgängen wurden die Teilnehmer zu verschiedenen Punkten im virtuellen Stadion teleportiert. Die Ergebnisse sprechen für erhöhte positive Affektivität (höhere Valenz- und Arousalratings, höhere Zygomaticusaktivität, mehr positiver emotionaler Ausdruck) durch angenehme im Vergleich zu neutralen Durchgängen. Reaktionen auf unangenehme im Vergleich zu neutralen Durchgängen waren widersprüchlich und sprechen für erhöhte positive und negative Affektivität. Unangenehme vs. neutrale Durchgänge führten zu niedrigeren Valenzratings, höheren Arousalratings, höherer Zygomaticusaktivität, etwas niedrigerer Corrugatoraktivität und mehr positivem und negativem emotionalen Ausdruck. Die Intensität emotionaler Reaktionen korrelierte mit dem Präsenzerleben in der virtuellen Realität. Um den Einfluss von Torschüssen allein vs. Torschüssen mit Trainerrückmeldung auf emotionale Reaktionen bei Gesunden besser zu verstehen, folgte in Experiment 2 nur auf 50 % aller Schüsse eine Trainerrückmeldung. Neutrale Durchgänge bestanden aus freien Schüssen über das Fußballfeld, auf die zu 50 % eine Trainerrückmeldung folgte. Schüsse mit Rückmeldung führten durchgängig zu stärkeren Valenz- und Arousalratings, höherer Zygomaticusaktivität, niedrigerer Corrugatoraktivität, und höheren Hautleitfähigkeits- reaktionen als Schüsse allein. Auch diese Ergebnisse sprechen für die Induktion positiver Emotionen durch angenehme Durchgänge, während die Induktion negativer Emotionen durch unangenehme Durchgänge uneindeutig scheint. Technische Verbesserungen der virtuellen Realität schlugen sich in höherem Präsenzerleben als in Experiment 1 nieder. Experiment 3 untersuchte emotionale Reaktionen von erwachsenen ADHS Patienten und gesunden Kontrollen nach Emotionsverarbeitung und reaktionsfokussierter Emotions- regulation. Versuchsteilnehmer absolvierten nacheinander ein vorgebliches Online-Ballspiel (Cyber Ball) welches negative Emotionen induzierte, und eine weiterentwickelte Version des virtuellen Elfmeterspiels. Während Cyber Ball wurden die Teilnehmer von zwei anderen Mitspielern in verschiedenen Versuchsblöcken entweder eingeschlossen oder ausgeschossen. Die Teilnehmer wurden in verschiedenen Versuchsblöcken instruiert, ihre Emotionen entweder deutlich zu zeigen, nicht zu regulieren, oder zu verbergen. Einige Ergebnisse sprechen für eine defizitäre Verarbeitung positiver Emotionen bei ADHS. Patienten berichteten niedrigeren positiven Affekt als Kontrollen während Cyber Ball, niedrigere Valenzratings als Kontrollen nach angenehmen Elfmeterdurchgängen, und zeigten niedrigere Zygomaticusaktivität als Kontrollen insbesondere während des Elfmeterschießens. Im Vergleich zu Kontrollen zeigten Patienten eine leicht verstärkte Verarbeitung von unangenehmen Ereignissen beim Cyber Ball (höhere Ratings von negativem Affekt v.a. nach Ausschluss), aber nicht beim Elfmeterschießen. Patienten zeigten eine niedrigere Baseline- Hautleitfähigkeit als Kontrollen, sowie beeinträchtigte Hautleitfähigkeitsmodulationen. Im Vergleich zu Kontrollen zeigten Patienten leicht erhöhten Ausdruck von positiven und negativen Emotionen. Emotionsregulationsanalysen zeigten keine bedeutenden Einschränkungen von ADHS vs. Kontrollen im Verändern ihrer emotionalen Reaktionen durch absichtliche reaktionsfokussierte Emotionsregulation. Außerdem gaben Patienten an, gewohnheitsmäßig sogar häufiger als Kontrollen adaptive Emotionsregulationsstrategien anzuwenden. Der Vergleich einer genetischen Hochrisikogruppe mit einer Niedrigrisikogruppe für ADHS über die gesamte Stichprobe zeigte ähnliche Ergebnisse wie die Analysen für Patienten vs. Kontrollen in der Modulation von Zygomaticusaktivität während der Emotionsverarbeitung, sowie in der Modulation emotionaler Reaktionen durch Emotionsregulation. Zusammenfassend lässt sich sagen, dass das virtuelle Elfmeterparadigma geeignet scheint, um positive, aber nicht negative Emotionen zu induzieren. Die Bedeutsamkeit von Präsenzerleben in einer virtuellen Realität für die Intensität von induzierten Emotionen konnte repliziert werden. ADHS Patienten zeigten Beeinträchtigungen v.a. in der Verarbeitung von positiven Emotionen. Abweichungen in der negativen emotionalen Reagibilität waren weniger eindeutig und sollten weiter untersucht werden. Die Ergebnisse deuten darauf hin, dass erwachsene ADHS Patienten im Vergleich zu gesunden Kontrollen eine niedrigere autonome Erregbarkeit bei Baseline und Defizite in der Erregbarkeitsmodulation zeigen. Es zeigten sich keine Defizite von ADHS in der absichtlichen Anwendung von ausdrucksfokussierter Emotionsregulation. KW - emotion processing KW - Emotionsverarbeitung KW - Emotionsregulation KW - ADHS KW - virtuelle Realität KW - virtual reality KW - adult ADHD KW - emotion regulation KW - Aufmerksamkeitsdefizit-Syndrom KW - Erwachsener KW - Gefühl Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-142064 ER - TY - JOUR A1 - Bechtle, Philip A1 - Camargo-Molina, José Eliel A1 - Desch, Klaus A1 - Dreiner, Herbert K. A1 - Hamer, Matthias A1 - Krämer, Michael A1 - O'Leary, Ben A1 - Porod, Werner A1 - Sarrazin, Björn A1 - Stefaniak, Tim A1 - Uhlenbrock, Mathias A1 - Wienemann, Peter T1 - Killing the cMSSM softly JF - The European Physical Journal C N2 - We investigate the constrained Minimal Supersymmetric Standard Model (cMSSM) in the light of constraining experimental and observational data from precision measurements, astrophysics, direct supersymmetry searches at the LHC and measurements of the properties of the Higgs boson, by means of a global fit using the program Fittino. As in previous studies, we find rather poor agreement of the best fit point with the global data. We also investigate the stability of the electro-weak vacuum in the preferred region of parameter space around the best fit point. We find that the vacuum is metastable, with a lifetime significantly longer than the age of the Universe. For the first time in a global fit of supersymmetry, we employ a consistent methodology to evaluate the goodness-of-fit of the cMSSM in a frequentist approach by deriving p values from large sets of toy experiments. We analyse analytically and quantitatively the impact of the choice of the observable set on the p value, and in particular its dilution when confronting the model with a large number of barely constraining measurements. Finally, for the preferred sets of observables, we obtain p values for the cMSSM below 10 %, i.e. we exclude the cMSSM as a model at the 90 % confidence level. KW - Dark Matter KW - Higgs Boson KW - Higgs Mass KW - Supersymmetry Breaking KW - Light Supersymmetric Particle Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-165045 VL - 76 IS - 96 ER - TY - JOUR A1 - Becker, Nils A1 - Kucharski, Robert A1 - Rössler, Wolfgang A1 - Maleszka, Ryszard T1 - Age‐dependent transcriptional and epigenomic responses to light exposure in the honey bee brain JF - FEBS Open Bio N2 - Light is a powerful environmental stimulus of special importance in social honey bees that undergo a behavioral transition from in-hive to outdoor foraging duties. Our previous work has shown that light exposure induces structural neuronal plasticity in the mushroom bodies (MBs), a brain center implicated in processing inputs from sensory modalities. Here, we extended these analyses to the molecular level to unravel light-induced transcriptomic and epigenomic changes in the honey bee brain. We have compared gene expression in brain compartments of 1- and 7-day-old light-exposed honey bees with age-matched dark-kept individuals. We have found a number of differentially expressed genes (DEGs), both novel and conserved, including several genes with reported roles in neuronal plasticity. Most of the DEGs show age-related changes in the amplitude of light-induced expression and are likely to be both developmentally and environmentally regulated. Some of the DEGs are either known to be methylated or are implicated in epigenetic processes suggesting that responses to light exposure are at least partly regulated at the epigenome level. Consistent with this idea light alters the DNA methylation pattern of bgm, one of the DEGs affected by light exposure, and the expression of microRNA miR-932. This confirms the usefulness of our approach to identify candidate genes for neuronal plasticity and provides evidence for the role of epigenetic processes in driving the molecular responses to visual stimulation. KW - DNA methylation KW - insect brain KW - light-induced gene expression KW - microRNA KW - neuronal plasticity Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-147080 VL - 6 IS - 7 ER - TY - JOUR A1 - Beer, Katharina A1 - Steffan-Dewenter, Ingolf A1 - Härtel, Stephan A1 - Helfrich-Förster, Charlotte T1 - A new device for monitoring individual activity rhythms of honey bees reveals critical effects of the social environment on behavior JF - Journal of Comparative Physiology A N2 - Chronobiological studies of individual activity rhythms in social insects can be constrained by the artificial isolation of individuals from their social context. We present a new experimental set-up that simultaneously measures the temperature rhythm in a queen-less but brood raising mini colony and the walking activity rhythms of singly kept honey bees that have indirect social contact with it. Our approach enables monitoring of individual bees in the social context of a mini colony under controlled laboratory conditions. In a pilot experiment, we show that social contact with the mini colony improves the survival of monitored young individuals and affects locomotor activity patterns of young and old bees. When exposed to conflicting Zeitgebers consisting of a light-dark (LD) cycle that is phase-delayed with respect to the mini colony rhythm, rhythms of young and old bees are socially synchronized with the mini colony rhythm, whereas isolated bees synchronize to the LD cycle. We conclude that the social environment is a stronger Zeitgeber than the LD cycle and that our new experimental set-up is well suited for studying the mechanisms of social entrainment in honey bees. KW - Social entrainment KW - Foragers KW - Nurses KW - Locomotor activity KW - Temperature rhythms Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-188030 VL - 202 IS - 8 ER - TY - JOUR A1 - Bekes, Inga A1 - Friedl, Thomas W. P. A1 - Köhler, Tanja A1 - Möbus, Volker A1 - Janni, Wolfgang A1 - Wöckel, Achim A1 - Wulff, Christine T1 - Does VEGF facilitate local tumor growth and spread into the abdominal cavity by suppressing endothelial cell adhesion, thus increasing vascular peritoneal permeability followed by ascites production in ovarian cancer? JF - Molecular Cancer N2 - Background Ovarian cancer is mostly associated with pathologically regulated permeability of peritoneal vessels, leading to ascites. Here, we investigated the molecular regulation of endothelial permeability by the vascular endothelial growth factor (VEGF) and both tight and adherens junction proteins (VE-cadherin and claudin 5) with regards to the tumor biology of different ovarian cancer types. Methods Serum and ascites samples before and after surgery, as well as peritoneal biopsies of 68 ovarian cancer patients and 20 healthy controls were collected. In serum and ascites VEGF protein was measured by ELISA. In peritoneal biopsies co-localization of VE-cadherin and claudin 5 was investigated using immunohistochemical dual staining. In addition, the gene expression of VE-cadherin and claudin 5 was quantified by Real-time PCR. Differences in VEGF levels, VE-cadherin and claudin 5 gene expression were analyzed in relation to various tumor characteristics (tumor stage, grading, histological subtypes, resection status after surgery) and then compared to controls. Furthermore, human primary ovarian cancer cells were co-cultured with human umbilical vein endothelial cells (HUVEC) and changes in VE-cadherin and claudin 5 were investigated after VEGF inhibition. Results VEGF was significantly increased in tumor patients in comparison to controls and accumulates in ascites. The highest VEGF levels were found in patients diagnosed with advanced tumor stages, with tumors of poor differentiation, or in the group of solid / cystic-solid tumors. Patients with residual tumor after operation showed significantly higher levels of VEGF both before and after surgery as compared to tumor-free resected patients. Results of an immunohistochemical double-staining experiment indicated co-localization of VE-cadherin and claudin 5 in the peritoneal vasculature. Compared to controls, expression of VE-cadherin and claudin 5 was significantly suppressed in peritoneal vessels of tumor patients, but there were no significant differences regarding VE-cadherin and claudin 5 expression in relation to different tumor characteristics. A significant positive correlation was found between VE-cadherin and claudin 5 expression. VEGF inhibition in vitro was associated with significant increase in VE-cadherin and claudin 5. Conclusions Our results indicate that increased peritoneal permeability in ovarian cancer is due to down-regulation of adhesion proteins via tumor derived VEGF. Advanced ovarian cancer with aggressive tumor biology may be associated with early dysregulation of vascular permeability leading to ascites. These patients may benefit from therapeutic VEGF inhibition. KW - ovarian cancer KW - vascular permeability KW - ascites KW - VEGF KW - claudin 5 KW - VE-cadherin Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-169298 VL - 15 IS - 13 ER - TY - THES A1 - Bellinger, Daniel T1 - Implementation of new reaction pathway determining methods and study of solvent effects on the excited state nature of perylene based dyes T1 - Implementierung neuer Reaktionspfad bestimmender Methoden und Untersuchung von Lösungsmitteleinflüssen auf die Natur der angeregten Zustände perylen-basierter Farbstoffe N2 - Two thematic complexes were addressed within this work. One part is related to improvements and new implementations into the CAST program package. Thereby the main focus laid on the delivery of a tool which can be used to characterize complex reactions and their mechanisms. But also within the new force field (FF) method (SAPT-FF) within the CAST program, several improvements were made. The second topic is related to the description of dye molecules and their spectral properties. The main focus within these studies was set on the influence of the environment on these properties. In the first topic improvements of the local acting NEB (nudged elastic band) methods were included and the number of available methods was extended. The initial pathway generation was improved by implementing the IDPP (image dependent pair potential) method and a new method was implemented for describing temperature dependent pathways. Additionally, improvements have been made to the optimization routines (global NEB). As a second part the Pathopt (PO) method was considerably improved. In the beginning of the work the original PO idea was used. In this approach one starts with a global optimization on one n-1 dimensional hyperplane which divides the reaction into two sub-areas for obtaining guesses of TSs (transition states). These found TS guesses were used to optimize to the ”true” TS. Starting from the optimized ones a relaxation to the next connected minima is done. This idea has been automatically implemented and extended to several number of hyperplanes. In this manner a group of pathsegments is obtained which needs to be connected, but within this work it was realized that such a procedure might be not very efficient. Therefore, a new strategy was implemented which is founded on the same constrained global optimization scheme (MCM) for which the user defines the number of hyperplanes generated. The number of such generated hyperplanes should be large enough 134 to describe the space between the concerning reactants in a sufficient way. The found minima are directly used to built up the reaction pathway. For this purpose a RMSD (root mean square deviation) criterion is used to walk along ways of minimal change from one to another hyperplane. To prove the implementations various test calculations were carried out and extensions included to prove the capabilities of the new strategy. Related to these tests a new strategy for applying the move steps in MCM (Monte Carlo with minimization) was realized which is also related to the question of the coordinates representation. We were able to show that the hopping steps in MCM can be improved by applying Cartesian steps in combination of random dihedral moves with respect to the constraint. In this way it was possible to show that a large variety of systems can be treated. An additional chapter shows the improvements of the SAPT-FF implementation and related test cases. It was possible to treat benzene dimer and cluster systems of different sizes consistently also in accordance with high level ab initio based approaches. Furthermore, we showed that the SAPT-FF with the right parameters outperforms the standard AMOEBA implementation which is the basis of the SAPT-FF implementation. In the last three chapters deal with the description of perlyene-based dyes. In the first smaller chapter ground state chemistry description of macro cycles of PBI (perylene bisimide) derivatives were investigated. Therefore, AFM (atomic force microscopy) based pictures were explained within our study. The methods to explain aggregation behavior in dependency of the ring size were MD simulations and configuration studies. The last two chapters deal with opto-electronic or photo-physical properties of PBI and PTCDA (perylene-3,4,9,10-tetracarboxylic dianhydride). In detail, we investigated the role of the environment and the aggregate or crystal surrounding by applying different models. In that way implicit and explicit solvation models, the size of aggregates and vibration motions were used. In the case of PBI the recent work is found on preliminary studies related to my bachelor thesis and extends it. It was shown that the direct influence of a polarizable surrounding, as well as explicit inclusion of solvent molecules on the overall description of the excitations and nature of the excited states is weaker as one might expect. However the inclusion of intra-molecular degrees of freedom showed a stronger influence on the state characteristics and can induce a change of the order of states within the dimer picture. For the PTCDA molecule the main focus was set on the description of the absorption spectrum of crystalline thin films. Related to this older works exist which already gave a description and assignment of the absorption band, but are based on different approaches compared to the one used in this work. We used the supermolecule ansatz, whereas the environment and different aggregate sizes were investigated. Within the dimer based approach we were able to show that using continuum solvation (IEFPCM/COSMO) based description for the environment the relative order of states remains unchanged. Similar to the PBI calculations the influence of the vibrational motions /distortions is larger. The simulation of the crystal environment by using QM/MM (quantum mechanics/molecular mechanics) approaches delivered that an asymmetric charge distribution might induce a localization of the excitation and a stronger mixing of states. For obtaining further insights we go beyond the dimer picture and aggregates of different sizes were used, whereas the simulations up to the octadecamer mono- and even dual-layer stack were carried out. Within these calculations it was shown that the H-coupling is dominating over a weaker J-coupling between different stacks. Additionally the calculations based on DFT (density functional theory) and semi-empirics showed that the lowest state in terms of energy are mostly of Frenkel type, whereas the higher lying states are CT ones which mix with embedded Frenkel type states. The first band of the absorption spectrum was explained by inclusion of vibrational motions within the stacks which induce an intensity gain of the first excited state. This intensity was not explainable by using the undistorted stacks. Also relaxations at the crystal surface might play a role, but are experimentally not explainable. N2 - In der Arbeit wurden zwei große Themenkomplexe bearbeitet. Zum einen wurden Verbesserungen und neue Methoden in CAST, unserem Entwicklungstool, implementiert. Hierbei geht es vor allem darum ein Werkzeug bereit zu stellen, mit dem es möglich ist Reaktionen genauer zu charakterisieren. Aber auch neue Beschreibungen innerhalb der Kraftfeldmethoden (SAPT-FF) wurden bereitgestellt bzw. erweitert. Der zweite Themenkomplex behandelt die Beschreibung von Farbstoffmolekülen und ihrer spektralen Eigenschaften. Insbesondere liegt in dieser Studie der Fokus auf Umgebungseinflüsse. Im ersten Abschnitt wurden Erweiterungen in den lokalen Methoden, die auf NEB (nudged elastic band) basieren, implementiert. Hier wurde zum einen das Spektrum an Methoden erweitert. So wurde der initiale Vorschlag für den Startpfad durch Implementierung der IDPP (image dependent pair potential) Methode verbessert. Des Weiteren wurde eine Methode zur temperaturabhängigen NEB Beschreibung integriert, die auf Maximierung des Fluxes beruht. Auch wurden Verbesserungen hinsichtlich der Optimierungsroutinen durchgeführt. Der wesentliche Teil im ersten Themenbereich beschäftigt sich mit der Verbesserung und Automatisierung von Pathopt (PO). Zu Beginn der Arbeit wurde die ursprüngliche Idee aufgegriffen. Hierbei ermittelt man Vorschläge für Übergangszustände (¨UZ) durch eine globale Optimierung mit Nebendbedingungen auf einer n-1 dimensionalen Hyperfläche, die den Reaktionsraum teilt. Diese ¨UZ bilden den Startpunkt, um mittels einer ”uphill” Optimierung hin zum ”wirklichen ÜZ” zu gelangen. Ausgehend von diesen wurde in die nächst verknüpften Minima relaxiert. Diese Idee wurde automatisiert und auf mehrere Hyperflächen ausgeweitet. So erhält man eine Schar an Pfadsegmenten, die verknüpft werden müssen. Im Laufe der Arbeit, stellte sich jedoch heraus, dass diese Vorgehensweise nicht sehr effizient ist und daher wurde eine neue Idee verwirklicht. Diese beruht wiederum auf der globalen Optimierung mittels Monte Carlo mit Minimierung und Nebenbedingungen auf einer vom Nutzer bestimmten Anzahl an n-1 dimensionalen Hyperflächen. Nun wählt man diese Anzahl entsprechend groß genug aus, um den Raum zwischen den Reaktanden zu beschreiben. Die so gefundenen Mininima auf den n-1 Hyperflächen werden für die direkte Pfaderzeugung genutzt. Dies geschieht mittels eines RMSD (root mean square deviation) Kriteriums, um so den Weg der geringsten Änderungen anhand der Hyperflächen zu wählen. Im Zuge der Implementierung der Methode wurden zahlreiche Testrechnungen und Methodenerweiterungen durchgeführt, um die Funktionalität zu überpüfen und zu verbessern. Diese Verbesserungen liegen zum Bsp. in den Sprungstrategien bzw. der Wahl des Koordinatensystems. Hier konnte gezeigt werden, dass eine Verbindung unterschiedlicher Strategien für die Durchführung des ”Hüpfens” in Monte Carlo zu entscheidenden Verbesserungen führt. Diese Verbesserung besteht in der Verknüpfung von Kartesischen Schritten und zufälliger Veränderungen der Diederwinkel im Rahmen der Nebenbedingungen. Mit Hilfe dieser Verbesserungen konnte eine Vielzahl von Systemen behandelt werden. Ein weiteres Kapitel beschreibt Verbesserungen zum SAPT-FF (FF=Kraftfeld). Testrechnungen zu strukturellen Eigenschaften von Benzol Clustern belegen die Genauigkeit der Ansätze. Auch wurde aufgezeigt, dass das SAPT-verbesserte AMOEBA Kraftfeld der Standard Parametrisierung überlegen ist. Die letzten drei Abschnitte dieser Arbeit behandeln Perylen-basierte Farbstoffe. In einem ersten kleinen Kapitel geht es um die Grundzustandseigenschaften von PBI (Perylenbisimide) Makrozyklen und Erklärung von AFM (Atomic Force Mycroscopy) Messungen. Hier konnten wir mittels MD-Simluation (Molekular Dynamik) und deren Analyse, sowie Beschreibungen unterschiedlicher Konfigurationen, das Aggregationsverhalten in Abhängigkeit der Ringgröße genauer beleuchten. In den beiden letzten Kapiteln geht es um die optoelektronischen Eigenschaften bzw. die photophysikalische Beschreibung von PBI und PTCDA (Perylen-3,4,9,10-Tetracarboxyl Dianhydrid). Im Genaueren wurde die Rolle der Umgebung in Aggregat und Kristall durch unterschiedliche methodische Ansätze untersucht. So wurden implizite Solvensmodelle und explizite Solvatation, Aggregatgröße und vibronische Freiheitsgrade untersucht. In den Arbeiten zum PBI konnte gezeigt werden, dass ein direkter Einfluss durch die Beschreibung mittels impliziter Solvatation, als auch expliziter Solvensmoleküle, auf die Lage der Zustände auch in Hinsicht auf deren Charakterisik nicht auftritt. Berücksichtigt man intra-molekulare Freiheitsgrade, so wird die Lage der Zustände deutlich stärker beeinflusst und sogar ein Wechsel der Zustände wird induziert. Im Fall von PTCDA lag vor allem die Beschreibung und Erklärung der Absorptionsspektren von kristallinem PTCDA im Fokus. Hierzu gibt es ¨altere Arbeiten, die bestimmte Zuordnungen der Banden und ihrer Übergänge postuliert haben. In dieser Arbeit sollte diese Beschreibung im Rahmen eines Supermolekül Ansatzes geklärt und weiter beschrieben werden, wobei Umgebungseinflüsse und auch Eigenschaften verschiedener Aggregate untersucht wurden. Im Dimer Bild konnten wir zeigen, dass die Umgebung, beschrieben durch Continuums Ansätze (IEFPCM/COSMO) die Lage der Zustände nicht beeinflusst und im Wesentlichen nur Zustände mit großer Oszillatorstärke stabilisiert werden. Ähnlich wie im Falle des PBIs hat die Berücksichtigung vibronischer Freiheitsgrade einen wesentlich größeren Einguss. Die Simulation der Kristallumgebung durch QM/MM-Ansätze (Quantenmechanik/Molekularmechanik) ergab, dass eine asymmetrische Ladungsverteilung zu einer Lokalisierung der Anregung und einem stärkeren Durchmischen der Zustände fuhrt. Für eine noch weitergehende Beschreibung wurde das Dimer Bild verlassen und unterschiedliche Aggregate, bis hin zum Oktadekamer im mono- bzw. zweifach-Lagen-Aggregat untersucht. Hier konnte gezeigt werden, dass die Kopplung im H-Aggregat die dominierende Rolle einnimmt und die J-Aggregat Kopplung vernachlässigt werden kann. Zudem zeigen die Rechnungen, die mittels DFT (Dichtefunktionaltheorie) und semi-empirischen Ansätzen durchgeführt wurden, dass die energetisch niedriger liegenden Zustände im wesentlichen Frenkel Charakter aufweisen während die energetisch höher liegenden Zustände CT (Charge Transfer) Charakter haben. Das Auftreten der ersten Bande im Absorptionsspektrum wurde zudem auf das Vorhandensein von möglichen Schwingungsanregungen (mehrere Moden) zurückgeführt, da diese zu einer Zunahme an Intensität des ersten angeregten Zustandes führen, die ohne Berücksichtigung nicht in diesem Maße erhalten wird. Auch könnten Oberflächenrelaxationen eine Rolle spielen, wobei diese experimentell nicht beobachtbar sind. KW - Globale Optimierung KW - Reaktionsmechanismus KW - Exziton KW - Reaktionspfadsuche KW - QM/MM KW - organische Halbleiter KW - Exzitonen Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-144435 ER - TY - JOUR A1 - Bemm, Felix A1 - Becker, Dirk A1 - Larisch, Christina A1 - Kreuzer, Ines A1 - Escalante-Perez, Maria A1 - Schulze, Waltraud X. A1 - Ankenbrand, Markus A1 - Van de Weyer, Anna-Lena A1 - Krol, Elzbieta A1 - Al-Rasheid, Khaled A. A1 - Mithöfer, Axel A1 - Weber, Andreas P. A1 - Schultz, Jörg A1 - Hedrich, Rainer T1 - Venus flytrap carnivorous lifestyle builds on herbivore defense strategies JF - Genome Research N2 - Although the concept of botanical carnivory has been known since Darwin's time, the molecular mechanisms that allow animal feeding remain unknown, primarily due to a complete lack of genomic information. Here, we show that the transcriptomic landscape of the Dionaea trap is dramatically shifted toward signal transduction and nutrient transport upon insect feeding, with touch hormone signaling and protein secretion prevailing. At the same time, a massive induction of general defense responses is accompanied by the repression of cell death-related genes/processes. We hypothesize that the carnivory syndrome of Dionaea evolved by exaptation of ancient defense pathways, replacing cell death with nutrient acquisition. KW - Dionaea-muscipula ellis KW - Plant utricularia-gibba KW - Programmed cell-death KW - Genomics data sets KW - RNA-SEQ data KW - Arabidopsis-thaliana KW - Jasmonate perception KW - Action potentials KW - Stress responses KW - Wonderful plants Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-188799 VL - 26 IS - 6 ER - TY - JOUR A1 - Ben Ami, Tal A1 - Tong, Yuehong A1 - Bhuiyan, Alauddin A1 - Huisingh, Carrie A1 - Ablonczy, Zsolt A1 - Ach, Thomas A1 - Curcio, Christine A. A1 - Smith, R. Theodore T1 - Spatial and Spectral Characterization of Human Retinal Pigment Epithelium Fluorophore Families by Ex Vivo Hyperspectral Autofluorescence Imaging JF - Translational Vision Science & Technology N2 - Purpose: Discovery of candidate spectra for abundant fluorophore families in human retinal pigment epithelium (RPE) by ex vivo hyperspectral imaging. Methods: Hyperspectral autofluorescence emission images were captured between 420 and 720 nm (10-nm intervals), at two excitation bands (436–460, 480–510 nm), from three locations (fovea, perifovea, near-periphery) in 20 normal RPE/Bruch's membrane (BrM) flatmounts. Mathematical factorization extracted a BrM spectrum (S0) and abundant lipofuscin/melanolipofuscin (LF/ML) spectra of RPE origin (S1, S2, S3) from each tissue. Results: Smooth spectra S1 to S3, with perinuclear localization consistent with LF/ML at all three retinal locations and both excitations in 14 eyes (84 datasets), were included in the analysis. The mean peak emissions of S0, S1, and S2 at λ\(_{ex}\) 436 nm were, respectively, 495 ± 14, 535 ± 17, and 576 ± 20 nm. S3 was generally trimodal, with peaks at either 580, 620, or 650 nm (peak mode, 650 nm). At λ\(_{ex}\) 480 nm, S0, S1, and S2 were red-shifted to 526 ± 9, 553 ± 10, and 588 ± 23 nm, and S3 was again trimodal (peak mode, 620 nm). S1 often split into two spectra, S1A and S1B. S3 strongly colocalized with melanin. There were no significant differences across age, sex, or retinal location. Conclusions: There appear to be at least three families of abundant RPE fluorophores that are ubiquitous across age, retinal location, and sex in this sample of healthy eyes. Further molecular characterization by imaging mass spectrometry and localization via super-resolution microscopy should elucidate normal and abnormal RPE physiology involving fluorophores. Translational Relevance: Our results help establish hyperspectral autofluorescence imaging of the human retinal pigment epithelium as a useful tool for investigating retinal health and disease. KW - spectral characterization KW - human retinal pigment epithelium KW - fluorophores KW - hyperspectral autofluorescence imaging Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-168328 VL - 5 IS - 3 ER - TY - JOUR A1 - Bening, C. A1 - Hamouda, K. A1 - Leyh, R. T1 - Sex differences in volume overload in skinned fibers JF - BMC Cardiovascular Disorders N2 - Background The impact of sex on cardiac morphology and function in chronic volume overload has been described in detail. However, the relation between sex and contractile properties at the actin-myosin level has not been well defined. Therefore, we evaluated the influence of sex on the contractile capacities of patients with chronic volume overload. Methods In 36 patients (18 males, 65 ± 9 years; 18 females, 65 ± 13 years) scheduled for elective mitral valve surgery due to severe mitral regurgitation (MR) with preserved left ventricular function, right auricle samples were obtained prior to extracorporal circulation. The fibers were prepared and skinned and exposed to a gradual increase in the calcium concentration (from pCa of 6.5–4.0) for calcium-induced force-developing measurements. Calcium sensitivity was also measured and recorded. Results The pCa-force relationship of the fibers obtained from males and females was significantly different, with the force values of the female fibers greater than those of male fibers at maximum calcium concentrations (pCa of 4.0: 3.6 ± 0.3 mN versus 3.2 ± 0.4 mN, p 0.02) and pCa of 4.5 2.6 ± 0.6 versus 2.0 ± 0.5, p 0.002). In contrast, the force values of female fibers were lower at mean calcium concentrations compared to those of male fibers (at 5.5 and pCa of 6.0: 1.0 ± 0.3 mN versus 1.2 ± 0.5 mN, p 0.04; 0.61 ± 0.05 versus 0.88 ± 0.09, p 0.04). Calcium sensitivity was observed at pCa of 5.0 in females and pCa of 4.5 in males. Conclusion This study demonstrated that female fibers from patients exposed to chronic volume overload developed higher force values at a given calcium concentration compared to fibers from male patients. We assume that female patients might tap the full force potential, which is required when exposed to the highest calcium concentrations in our experimental cycle. The calcium sensitivity among genders was significantly different, with the results suggesting that males have higher calcium sensitivity and might compensate for lower force values at maximal calcium concentrations by a higher affinity for calcium. Hence, female patients with MR seem to work more “energy efficient”. KW - calcium sensitivity KW - pCa KW - force relationship KW - skinned fibers Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-147896 VL - 16 IS - 197 ER - TY - JOUR A1 - Benoit, Joshua B. A1 - Adelman, Zach N. A1 - Reinhardt, Klaus A1 - Dolan, Amanda A1 - Poelchau, Monica A1 - Jennings, Emily C. A1 - Szuter, Elise M. A1 - Hagan, Richard W. A1 - Gujar, Hemant A1 - Shukla, Jayendra Nath A1 - Zhu, Fang A1 - Mohan, M. A1 - Nelson, David R. A1 - Rosendale, Andrew J. A1 - Derst, Christian A1 - Resnik, Valentina A1 - Wernig, Sebastian A1 - Menegazzi, Pamela A1 - Wegener, Christian A1 - Peschel, Nicolai A1 - Hendershot, Jacob M. A1 - Blenau, Wolfgang A1 - Predel, Reinhard A1 - Johnston, Paul R. A1 - Ioannidis, Panagiotis A1 - Waterhouse, Robert M. A1 - Nauen, Ralf A1 - Schorn, Corinna A1 - Ott, Mark-Christoph A1 - Maiwald, Frank A1 - Johnston, J. Spencer A1 - Gondhalekar, Ameya D. A1 - Scharf, Michael E. A1 - Raje, Kapil R. A1 - Hottel, Benjamin A. A1 - Armisén, David A1 - Crumière, Antonin Jean Johan A1 - Refki, Peter Nagui A1 - Santos, Maria Emilia A1 - Sghaier, Essia A1 - Viala, Sèverine A1 - Khila, Abderrahman A1 - Ahn, Seung-Joon A1 - Childers, Christopher A1 - Lee, Chien-Yueh A1 - Lin, Han A1 - Hughes, Daniel S.T. A1 - Duncan, Elizabeth J. A1 - Murali, Shwetha C. A1 - Qu, Jiaxin A1 - Dugan, Shannon A1 - Lee, Sandra L. A1 - Chao, Hsu A1 - Dinh, Huyen A1 - Han, Yi A1 - Doddapaneni, Harshavardhan A1 - Worley, Kim C. A1 - Muzny, Donna M. A1 - Wheeler, David A1 - Panfilio, Kristen A. A1 - Jentzsch, Iris M. Vargas A1 - Jentzsch, IMV A1 - Vargo, Edward L. A1 - Booth, Warren A1 - Friedrich, Markus A1 - Weirauch, Matthew T. A1 - Anderson, Michelle A.E. A1 - Jones, Jeffery W. A1 - Mittapalli, Omprakash A1 - Zhao, Chaoyang A1 - Zhou, Jing-Jiang A1 - Evans, Jay D. A1 - Attardo, Geoffrey M. A1 - Robertson, Hugh M. A1 - Zdobnov, Evgeny M. A1 - Ribeiro, Jose M.C. A1 - Gibbs, Richard A. A1 - Werren, John H. A1 - Palli, Subba R. A1 - Schal, Coby A1 - Richards, Stephen T1 - Unique features of a global human ectoparasite identified through sequencing of the bed bug genome JF - Nature Communications N2 - The bed bug, Cimex lectularius, has re-established itself as a ubiquitous human ectoparasite throughout much of the world during the past two decades. This global resurgence is likely linked to increased international travel and commerce in addition to widespread insecticide resistance. Analyses of the C. lectularius sequenced genome (650 Mb) and 14,220 predicted protein-coding genes provide a comprehensive representation of genes that are linked to traumatic insemination, a reduced chemosensory repertoire of genes related to obligate hematophagy, host–symbiont interactions, and several mechanisms of insecticide resistance. In addition, we document the presence of multiple putative lateral gene transfer events. Genome sequencing and annotation establish a solid foundation for future research on mechanisms of insecticide resistance, human–bed bug and symbiont–bed bug associations, and unique features of bed bug biology that contribute to the unprecedented success of C. lectularius as a human ectoparasite. KW - human ectoparasite KW - bed bug KW - Cimex lectularius KW - genome Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-166221 VL - 7 IS - 10165 ER - TY - JOUR A1 - Berges, Carsten A1 - Kerkau, Thomas A1 - Werner, Sandra A1 - Wolf, Nelli A1 - Winter, Nadine A1 - Hünig, Thomas A1 - Einsele, Hermann A1 - Topp, Max S. A1 - Beyersdorf, Niklas T1 - Hsp90 inhibition ameliorates CD4\(^{+}\) T cell-mediated acute Graft versus Host disease in mice JF - Immunity, Inflammation and Disease N2 - Introduction: For many patients with leukemia only allogeneic bone marrow transplantion provides a chance of cure. Co‐transplanted mature donor T cells mediate the desired Graft versus Tumor (GvT) effect required to destroy residual leukemic cells. The donor T cells very often, however, also attack healthy tissue of the patient inducing acute Graft versus Host Disease (aGvHD)—a potentially life‐threatening complication. Methods: Therefore, we used the well established C57BL/6 into BALB/c mouse aGvHD model to evaluate whether pharmacological inhibition of heat shock protein 90 (Hsp90) would protect the mice from aGvHD. Results: Treatment of the BALB/c recipient mice from day 0 to +2 after allogeneic CD4\(^{+}\) T cell transplantation with the Hsp90 inhibitor 17‐(dimethylaminoethylamino)‐17‐demethoxygeldanamycin (DMAG) partially protected the mice from aGvHD. DMAG treatment was, however, insufficient to prolong overall survival of leukemia‐bearing mice after transplantation of allogeneic CD4\(^{+}\) and CD8\(^{+}\) T cells. Ex vivo analyses and in vitro experiments revealed that DMAG primarily inhibits conventional CD4\(^{+}\) T cells with a relative resistance of CD4\(^{+}\) regulatory and CD8\(^{+}\) T cells toward Hsp90 inhibition. Conclusions: Our data, thus, suggest that Hsp90 inhibition might constitute a novel approach to reduce aGvHD in patients without abrogating the desired GvT effect. KW - Hsp90 KW - leukemia KW - acute Graft versus Host Disease KW - Graft versus Tumor Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-168318 VL - 4 IS - 4 ER - TY - JOUR A1 - Bernt, Alexander A1 - Rangrez, Ashraf Y. A1 - Eden, Matthias A1 - Jungmann, Andreas A1 - Katz, Sylvia A1 - Rohr, Claudia A1 - Müller, Oliver J. A1 - Katus, Hugo A. A1 - Sossalla, Samuel T. A1 - Williams, Tatjana A1 - Ritter, Oliver A1 - Frank, Derk A1 - Frey, Norbert T1 - Sumoylation-independent activation of Calcineurin-NFAT-signaling via SUMO2 mediates cardiomyocyte hypertrophy JF - Scientific Reports N2 - The objective of this study was to identify unknown modulators of Calcineurin (Cn)-NFAT signaling. Measurement of NFAT reporter driven luciferase activity was therefore utilized to screen a human cardiac cDNA-library (~10\(^{7}\) primary clones) in C2C12 cells through serial dilutions until single clones could be identified. This extensive screening strategy culminated in the identification of SUMO2 as a most efficient Cn-NFAT activator. SUMO2-mediated activation of Cn-NFAT signaling in cardiomyocytes translated into a hypertrophic phenotype. Prohypertrophic effects were also observed in mice expressing SUMO2 in the heart using AAV9 (Adeno-associated virus), complementing the in vitro findings. In addition, increased SUMO2-mediated sumoylation in human cardiomyopathy patients and in mouse models of cardiomyopathy were observed. To decipher the underlying mechanism, we generated a sumoylation-deficient SUMO2 mutant (ΔGG). Surprisingly, ΔGG replicated Cn-NFAT-activation and the prohypertrophic effects of native SUMO2, both in vitro and in vivo, suggesting a sumoylation-independent mechanism. Finally, we discerned a direct interaction between SUMO2 and CnA, which promotes CnA nuclear localization. In conclusion, we identified SUMO2 as a novel activator of Cn-NFAT signaling in cardiomyocytes. In broader terms, these findings reveal an unexpected role for SUMO2 in cardiac hypertrophy and cardiomyopathy, which may open the possibility for therapeutic manipulation of this pathway. KW - Calcineurin-NFATsignaling KW - activation KW - SUMO2 KW - cardiac hypertrophy Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-167525 VL - 6 IS - 35758 ER - TY - JOUR A1 - Bert, Bettina A1 - Chmielewska, Justyna A1 - Bergmann, Sven A1 - Busch, Maximilian A1 - Driever, Wolfgang A1 - Finger-Baier, Karin A1 - Hößler, Johanna A1 - Köhler, Almut A1 - Leich, Nora A1 - Misgeld, Thomas A1 - Nöldner, Torsten A1 - Reiher, Annegret A1 - Schartl, Manfred A1 - Seebach-Sproedt, Anja A1 - Thumberger, Thomas A1 - Schönfelder, Gilbert A1 - Grune, Barbara T1 - Considerations for a European animal welfare standard to evaluate adverse phenotypes in teleost fish JF - The EMBO Journal N2 - No abstract available. KW - Danio-rerio KW - Zebrafish KW - Pain Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-188783 VL - 35 IS - 11 ER - TY - THES A1 - Bertho, Sylvain T1 - Biochemical and molecular characterization of an original master sex determining gene in Salmonids T1 - Biochemische und molekulare Charakterisierung des Mastergens bei der Sex-bestimmung in Salmoniden N2 - Sexual development is a fundamental and versatile process that shapes animal morphology, physiology and behavior. The underlying developmental process is composed of the sex determination and the sex differentiation. Sex determination mechanisms are extremely labile among taxa. The initial triggers of the sex determination process are often genetics called sex determining genes. These genes are expressed in the bipotential gonad and tilt the balance to a developmental program allowing the differentiation of either a testis or an ovary. Fish represent a large and fascinating vertebrate group to study both sex determination and sex differentiation mechanisms. To date, among the known sex determining genes, three gene families namely sox, dmrt and TGF-β factors govern this developmental program. As exception to this rule, sdY “sexually dimorphic on the Y” does not belong to one of these families as it comes from the duplication / evolution of an ancestor gene related to immunity, i.e., the interferon related factor 9, irf9. sdY is the master sex determining gene in salmonids, a group of fishes that include species such as rainbow trout and Atlantic salmon. The present study was aimed to firstly characterize the features of SdY protein. Results indicate that SdY is predominantly localized in the cytoplasm tested in various fish and mammalian cell lines and confirmed by different methods. Predictive in silico analysis revealed that SdY is composed of a β-sandwich core surrounded by three α-helices as well specific characteristics conferring a putative protein-protein interaction site. Secondly, the study was aimed to understand how SdY could trigger testicular differentiation. SdY is a truncated divergent version of Irf9 that has a conserved protein-protein domain but lost the DNA interaction domain of its ancestor gene. It was then hypothesized that SdY could initiate testicular differentiation by protein-protein interactions. To evaluate this we first conducted a yeast-two-hybrid screen that revealed a high proportion of transcription factors including fox proteins. Using various biochemical and cellular methods we confirm an interaction between SdY and Foxl2, a major transcription factor involved in ovarian differentiation and identity maintenance. Interestingly, the interaction of SdY with Foxl2 leads to nuclear translocation of SdY from the cytoplasm. Furthermore, this SdY translocation mechanism was found to be specific to fish Foxl2 and to a lesser extend Foxl3 and not other Fox proteins or mammalian FoxL2. In addition, we found that this interaction allows the stabilization of SdY and prevents its degradation. Finally, to better decipher SdY action we used as a model a mutated version of SdY that was identified in XY females of Chinook salmon natural population. Results show that this mutation induces a local conformation defect obviously leading to a misfolded protein and a quick degradation. Moreover, the mutated version compromised the interaction with Foxl2 defining a minimal threshold to induce testicular differentiation. Altogether results from my thesis propose that SdY would trigger testicular differentiation in salmonids by preventing Foxl2 to promote ovarian differentiation. Further research should be now carried out on how this interaction of SdY and Foxl2 acts in-vivo. N2 - Le développement du sexe est un processus fondamental et versatile qui forme la morphologie, la physiologie et le comportement des animaux. Le processus de développement sous-jacent est composé de la détermination et de la différentiation du sexe. Les mécanismes de détermination du sexe sont extrêment labile parmi les taxons. Les signaux initiaux du processus de détermination du sexe sont souvent génétiques et nommés gènes de détermination du sexe. Ces gènes sont exprimés dans la gonade bipotente et font pencher l’équilibre vers un programme de développement permettant la formation soit d’un testicule soit d’un ovaire. Les poissons représentent un large et fascinant groupe de vertébrés pour étudier les processus de détermination et de différentiation du sexe. A l’heure actuelle, parmi les gènes de détermination connus, trois familles de gènes nommément sox, dmrt and les facteurs TGF-β gouvernent ce processus de développement. Comme exception à cette règle, sdY « sexually dimorphic on the Y » n’appartient à aucune de ces familles puisqu’il provient d’une duplication/évolution d’un gène ancestral de l’immunité, c’est-à-dire d’un facteur lié à l’interféron, irf9. sdY est le gène maître de la détermination du sexe chez les salmonidés, un groupe de poissons incluant des espèces tel que la truite arc-en-ciel et le saumon Altantique. L’étude présentée avait pour but de premièrement caractériser les propriétés de la protéine SdY. Les résultats indiquent que SdY est localisée de façon prédominante dans le cytoplasme testés dans diverses cellules de poissons et de mammifères et confirmé par des différentes méthodes. Une analyse in silico prédictive a révélé que SdY est composé d’un core β-sandwich entouré par trois hélices-α ainsi que des caractéristiques lui conférant un site d’interaction protéine-protéine. Deuxièment, l’étude avait pour but de comprendre comment SdY pouvait entraîner la différentiation testiculaire. SdY est une version tronquée divergente de Irf9 qui a conservé le domaine protéine-protéine mais a perdu le domaine d’interaction à l’ADN présent dans le gène ancestral. Il a été proposé que SdY entraîne la différentiation testiculaire par interaction(s) protéine-protéine. Afin d’évaluer cette hypothèse, un crible double-hybride en système levure a révélé une forte proportion de facteurs de transcription incluant les protéines fox. En utilisant de nombreuses méthodes au niveau cellulaire et biochimique, nous avons confirmé une interaction entre SdY et Foxl2, un facteur majeur impliqué dans la différentiation ovarienne et gardien de son identité. De façon intéressante, l’interaction de SdY avec Foxl2 conduit à une translocation nucléaire de SdY à partir du cytoplasme. De plus, le mécanisme de translocation de SdY est spécifique à la protéine Foxl2 et dans une moindre mesure à Foxl3 parmi les protéines Fox de poissons ou bien des protéines FoxL2 de mammifères. Puis, nous avons montré que cette interaction permet la stabilisation de SdY et empêche sa dégradation. Enfin, pour mieux décrypter l’action de SdY, nous avons utilisé comme modèle une version mutée qui a été identifiée dans une population naturelle de saumon Chinook avec des individus XY femelles. Les résultats montrent que la mutation induit un défaut de conformation local menant à une protéine mal-repliée et à sa dégradation. De plus, la version mutée compromet l’interaction avec Foxl2 définissant un seuil minimal d’induction de la différentiation testiculaire. Les résultats de ma thèse pris dans leur ensemble proposent que SdY pourrait entraîner la différentiation testiculaire chez les salmonidés en empêchant Foxl2 d’induire la différentiation ovarienne. Les recherches doivent se poursuivre dans le but de comprendre comment l’interaction SdY avec Foxl2 fonctionne in vivo. N2 - Sexuelle Entwicklung ist ein grundlegender und vielfältiger Prozess, der die Morphologie, Physiologie und das Verhalten von Tieren gestaltet. Der zugrundeliegende Entwicklungsprozess besteht aus der Geschlechtsbestimmung und der Geschlechtsdifferenzierung. Die Mechanismen der Geschlechtsbestimmung sind sehr instabil zwischen verschiedenen Arten. Die Auslöser des Prozesses der Geschlechtsbestimmung sind oft genetischen Ursprungs wie geschlechtsbestimmende Gene. Diese Gene werden in den bipotentialen Gonaden exprimiert und steuern die Balance eines entwicklungsgemäßen Programms, das die Differenzierung zum Testis oder Ovar erlaubt. Fische repräsentieren eine umfangreiche und faszinierende Gruppe von Vertebraten, um die Mechanismen der Geschlechtsbestimmung und –differenzierung zu untersuchen. Bislang ist bekannt, dass –unter den bekannten geschlechtsbestimmenden Genen- die drei Gen-Familien sox, dmrt und die TGFß-Faktoren dieses Entwicklungsprogramm steuern. Als Ausnahme von dieser Regel ist sdY „sexually dimorphic on the Y“ keiner dieser Familien zugehörig da es von der Duplikation / Evolution eines Vorgänger-Gens, das mit Immunität wie z.B. interferon related factor9, irf9, in Verbindung steht, herrührt. sdY ist das Mastergen der Geschlechtsbestimmung in Salmoniden, die als Gruppe von Fischen Arten wie die Regenbogenforelle und den Atlantischen Lachs umfassen. Das Ziel der vorliegenden Arbeit war es zunächst die Eigenschaften des SdY Proteins zu charakterisieren. Die Ergebnisse zeigen, dass SdY vor allem im Zytoplasma lokalisiert ist. Dies wurde in verschiedenen Fischen und Säugetier Zelllinien untersucht und mit Hilfe verschiedener Methoden bestätigt. Prädiktive in silico Analysen zeigten, dass SdY aus einem ß-sandwich Kern besteht, der von drei α-Helices umgeben ist sowie spezifischen Eigenschaften für eine putative Protein-Protein Interaktion Stelle. Das zweite Ziel der vorliegenden Arbeit war es, zu verstehen, wie SdY die testikuläre Differenzierung auslösen könnte. SdY ist eine verkürzte, divergente Version von Irf9, das eine konservierte Protein-Protein Domäne aufweist, jedoch seine DNA Interaktion Domäne a seines Vorläufer Gens verloren hat. Daher wurde angenommen, dass SdY die testikuläre Differenzierung durch Protein-Protein Interaktion initiieren könnte. Um diese Hypothese zu bestätigen führten wir zuerst einen Yeast Two-Hybrid Screen durch, der einen hohen Anteil an Transkriptionsfaktoren darunter fox Proteine zeigte. Unter Einsatz verschiedener biochemischer und zellulärer Methoden bestätigten wir eine Interaktion zwischen SdY und Foxl2, einem wesentlichen Transkriptionsfaktor, der in die Differenzierung und die Erhaltung der Identität der Ovarien involviert ist. Interessanterweise führt die Interaktion von SdY mit Foxl2 zu einer nukleären Translokation von SdY aus dem Zytoplasma. Außerdem wurde festgestellt, dass dieser SdY Translokations-Mechanismus für das Fisch Foxl2 und in einem geringerem Maße für Foxl3 spezifisch ist aber nicht für andere Fox Proteine oder Säuger FoxL2. Des Weiteren haben wir herausgefunden, dass diese Interaktion die Stabilisierung von SdY ermöglicht und sein Abbau verhindert. Zuletzt haben wir ein Modell einer mutierten Version von SdY benutzt, die in XY Weibchen der natürlichen Population der Königslachse identifiziert wurde, um die Wirkung von SdY besser zu entschlüsseln. Die Ergebnisse zeigen, dass diese Mutation einen lokalen Konformationsdefekt verursacht, der zu fehlgefalteten Proteinen und einem raschen Abbau führt. Darüber hinaus beeinträchtigt die mutierte Version die Interaktion mit FoxL2 und definiert einen minimalen Grenzwert, um die testikuläre Differenzierung zu induzieren. Insgesamt deuten die Ergebnisse meiner Dissertation darauf hin, dass SdY die testikuläre Differenzierung in Salmoniden auslöst, indem es verhindert, dass Foxl2 die Differenzierung der Ovarien fördert. In Zukunft soll erforscht werden, wie sich die Interaktion von SdY und Foxl2 in-vivo auswirkt. KW - Fish Sex determination KW - gonad development KW - SdY KW - salmonids KW - Lachsartige KW - Geschlechtsdifferenzierung KW - Molekulargenetik Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-139130 ER - TY - JOUR A1 - Beykan, Seval A1 - Dam, Jan S. A1 - Eberlein, Uta A1 - Kaufmann, Jens A1 - Kjærgaard, Benedict A1 - Jødal, Lars A1 - Bouterfa, Hakim A1 - Bejot, Romain A1 - Lassmann, Michael A1 - Jensen, Svend Borup T1 - \(^{177}\)Lu-OPS201 targeting somatostatin receptors: in vivo biodistribution and dosimetry in a pig model JF - EJNMMI Research N2 - Background \(^{177}\)Lu is used in peptide receptor radionuclide therapies for the treatment of neuroendocrine tumors. Based on the recent literature, SST2 antagonists are superior to agonists in tumor uptake. The compound OPS201 is the novel somatostatin antagonist showing the highest SST2 affinity. The aim of this study was to measure the in vivo biodistribution and dosimetry of \(^{177}\)Lu-OPS201 in five anesthetized Danish Landrace pigs as an appropriate substitute for humans to quantitatively assess the absorbed doses for future clinical applications. Results \(^{177}\)Lu-OPS201 was obtained with a specific activity ranging from 10 to 17 MBq/μg. Prior to administration, the radiochemical purity was measured as s > 99.7 % in all cases. After injection, fast clearance of the compound from the blood stream was observed. Less than 5 % of the injected activity was presented in blood 10 min after injection. A series of SPECT/CT and whole-body scans conducted until 10 days after intravenous injection showed uptake mostly in the liver, spine, and kidneys. There was no visible uptake in the spleen. Blood samples were taken to determine the time-activity curve in the blood. Time-activity curves and time-integrated activity coefficients were calculated for the organs showing visible uptake. Based on these data, the absorbed organ dose coefficients for a 70-kg patient were calculated with OLINDA/EXM. For humans after an injection of 5 GBq \(^{177}\)Lu-OPS201, the highest predicted absorbed doses are obtained for the kidneys (13.7 Gy), the osteogenic cells (3.9 Gy), the urinary bladder wall (1.8 Gy), and the liver (1.0 Gy). No metabolites of 177Lu-OPS201 were found by radio HPLC analysis. None of the absorbed doses calculated will exceed organ toxicity levels. Conclusions The \(^{177}\)Lu-OPS201 was well tolerated and caused no abnormal physiological or behavioral signs. In vivo distributions and absorbed doses of pigs are comparable to those observed in other publications. According to the biodistribution data in pigs, presented in this work, the expected radiation exposure in humans will be within the acceptable range. KW - lutetium-177 KW - JR11 KW - antagonist KW - dosimetry KW - neuroendocrine tumor (NET) KW - OPS201 KW - pig model KW - PRRT Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-146888 VL - 6 IS - 50 ER - TY - JOUR A1 - Bhavsar, Shefalee K. A1 - Singh, Yogesh A1 - Sharma, Piyush A1 - Khairnar, Vishal A1 - Hosseinzadeh, Zohreh A1 - Zhang, Shaqiu A1 - Palmada, Monica A1 - Sabolic, Ivan A1 - Koepsell, Hermann A1 - Lang, Karl S. A1 - Lang, Philipp A. A1 - Lang, Florian T1 - Expression of JAK3 Sensitive Na\(^+\) Coupled Glucose Carrier SGLT1 in Activated Cytotoxic T Lymphocytes JF - Cellular Physiology and Biochemistry N2 - Background: Similar to tumor cells, activated T-lymphocytes generate ATP mainly by glycolytic degradation of glucose. Lymphocyte glucose uptake involves non-concentrative glucose carriers of the GLUT family. In contrast to GLUT isoforms, Na+-coupled glucose-carrier SGLT1 accumulates glucose against glucose gradients and is effective at low extracellular glucose concentrations. The present study explored expression and regulation of SGLT1 in activated murine splenic cytotoxic T cells (CTLs) and human Jurkat T cells. Methods: FACS analysis, immunofluorescence, confocal microscopy, chemiluminescence and Western blotting were employed to estimate SGLT1 expression, function and regulation in lymphocytes, as well as dual electrode voltage clamp in SGLT1 ± JAK3 expressing Xenopus oocytes to quantify the effect of janus kinase3 (JAK3) on SGLT1 function. Results: SGLT1 is expressed in murine CTLs and also in human Jurkat T cells. 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose uptake was significantly decreased by SGLT1-blocker phloridzin (0.2 mM) and by pharmacological inhibition of JAK3 with WHI-P131 (156 µM), WHI-P154 (11.2 µM) and JAK3 inhibitor VI (0.5 µM). Electrogenic glucose transport (Iglucose) in Xenopus oocytes expressing human SGLT1 was increased by additional expression of human wild type JAK3, active A568VJAK3 but not inactive K851AJAK3. Coexpression of JAK3 enhanced the maximal transport rate without significantly modifying affinity of the carrier. Iglucose in SGLT1+JAK3 expressing oocytes was significantly decreased by WHI-P154 (11.2 µM). JAK3 increased the SGLT1 protein abundance in the cell membrane. Inhibition of carrier insertion by brefeldin A (5 µM) in SGLT1+JAK3 expressing oocytes resulted in a decline of Iglucose, which was similar in presence and absence of JAK3. Conclusions: SGLT1 is expressed in murine cytotoxic T cells and human Jurkat T cells and significantly contributes to glucose uptake in those cells post activation. JAK3 up-regulates SGLT1 activity by increasing the carrier protein abundance in the cell membrane, an effect enforcing cellular glucose uptake into activated lymphocytes and thus contributing to the immune response. KW - tumor cell KW - Cytotoxic T lymphocytes KW - Glucose uptake KW - Jurkat T cells KW - Energy depletion KW - Janus kinase Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-164900 VL - 39 IS - 3 ER - TY - JOUR A1 - Bialas, David A1 - Zitzler-Kunkel, André A1 - Kirchner, Eva A1 - Schmidt, David A1 - Würthner, Frank T1 - Structural and quantum chemical analysis of exciton coupling in homo- and heteroaggregate stacks of merocyanines JF - Nature Communications N2 - Exciton coupling is of fundamental importance and determines functional properties of organic dyes in (opto-)electronic and photovoltaic devices. Here we show that strong exciton coupling is not limited to the situation of equal chromophores as often assumed. Quadruple dye stacks were obtained from two bis(merocyanine) dyes with same or different chromophores, respectively, which dimerize in less-polar solvents resulting in the respective homo- and heteroaggregates. The structures of the quadruple dye stacks were assigned by NMR techniques and unambiguously confirmed by single-crystal X-ray analysis. The heteroaggregate stack formed from the bis(merocyanine) bearing two different chromophores exhibits remarkably different ultraviolet/vis absorption bands compared with those of the homoaggregate of the bis(merocyanine) comprising two identical chromophores. Quantum chemical analysis based on an extension of Kasha’s exciton theory appropriately describes the absorption properties of both types of stacks revealing strong exciton coupling also between different chromophores within the heteroaggregate. KW - exciton coupling KW - merocyanines KW - quantum chemical analysis Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-170200 VL - 7 ER - TY - JOUR A1 - Biedermann, B. A1 - Billoni, M. A1 - Denner, A. A1 - Dittmaier, S. A1 - Hofer, L. A1 - Jäger, B. A1 - Salfelder, L. T1 - Next-to-leading-order electroweak corrections to pp -> W\(^{+}\)W\(^{-}\) -> 4 leptons at the LHC JF - JOURNAL OF HIGH ENERGY PHYSICS N2 - We present results of the first calculation of next-to-leading-order electroweak corrections to W-boson pair production at the LHC that fully takes into account leptonic W-boson decays and off-shell effects. Employing realistic event selections, we discuss the corrections in situations that are typical for the study of W-boson pairs as a signal process or of Higgs-boson decays H → WW∗, to which W-boson pair production represents an irreducible background. In particular, we compare the full off-shell results, obtained treating the W-boson resonances in the complex-mass scheme, to previous results in the so-called double-pole approximation, which is based on an expansion of the loop amplitudes about the W resonance poles. At small and intermediate scales, i.e. in particular in angular and rapidity distributions, the two approaches show the expected agreement at the level of fractions of a percent, but larger differences appear in the TeV range. For transverse-momentum distributions, the differences can even exceed the 10% level in the TeV range where “background diagrams” with one instead of two resonant W bosons gain in importance because of recoil effects. KW - NLO Computations KW - W-boson KW - LHC Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-167790 VL - 06 IS - 065 ER - TY - JOUR A1 - Biehl, Stefanie C. A1 - Merz, Christian J. A1 - Dresler, Thomas A1 - Heupel, Julia A1 - Reichert, Susanne A1 - Jacob, Christian P. A1 - Deckert, Jürgen A1 - Herrmann, Martin J. T1 - Increase or Decrease of fMRI Activity in Adult Attention Deficit/ Hyperactivity Disorder: Does It Depend on Task Difficulty? JF - International Journal of Neuropsychopharmacology N2 - Background: Attention deficit/hyperactivity disorder has been shown to affect working memory, and fMRI studies in children and adolescents with attention deficit/hyperactivity disorder report hypoactivation in task-related attentional networks. However, studies with adult attention deficit/hyperactivity disorder patients addressing this issue as well as the effects of clinically valid methylphenidate treatment are scarce. This study contributes to closing this gap. Methods: Thirty-five adult patients were randomized to 6 weeks of double-blind placebo or methylphenidate treatment. Patients completed an fMRI n-back working memory task both before and after the assigned treatment, and matched healthy controls were tested and compared to the untreated patients. Results: There were no whole-brain differences between any of the groups. However, when specified regions of interest were investigated, the patient group showed enhanced BOLD responses in dorsal and ventral areas before treatment. This increase was correlated with performance across all participants and with attention deficit/hyperactivity disorder symptoms in the patient group. Furthermore, we found an effect of treatment in the right superior frontal gyrus, with methylphenidate-treated patients exhibiting increased activation, which was absent in the placebo-treated patients. Conclusions: Our results indicate distinct activation differences between untreated adult attention deficit/hyperactivity disorder patients and matched healthy controls during a working memory task. These differences might reflect compensatory efforts by the patients, who are performing at the same level as the healthy controls. We furthermore found a positive effect of methylphenidate on the activation of a frontal region of interest. These observations contribute to a more thorough understanding of adult attention deficit/hyperactivity disorder and provide impulses for the evaluation of therapy-related changes. KW - working memory KW - clinical trial KW - child memory KW - short-term methylphenidate brain KW - methylphenidate KW - adult attention deficit/hyperactivity disorder KW - fMRI KW - functional magnetic resonance imaging Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-147551 VL - 19 IS - 10 ER - TY - JOUR A1 - Biscotti, Maria Assunta A1 - Gerdol, Marco A1 - Canapa, Adriana A1 - Forconi, Mariko A1 - Olmo, Ettore A1 - Pallavicini, Alberto A1 - Barucca, Marco A1 - Schartl, Manfred T1 - The Lungfish Transcriptome: A Glimpse into Molecular Evolution Events at the Transition from Water to Land JF - Scientific Reports N2 - Lungfish and coelacanths are the only living sarcopterygian fish. The phylogenetic relationship of lungfish to the last common ancestor of tetrapods and their close morphological similarity to their fossil ancestors make this species uniquely interesting. However their genome size, the largest among vertebrates, is hampering the generation of a whole genome sequence. To provide a partial solution to the problem, a high-coverage lungfish reference transcriptome was generated and assembled. The present findings indicate that lungfish, not coelacanths, are the closest relatives to land-adapted vertebrates. Whereas protein-coding genes evolve at a very slow rate, possibly reflecting a “living fossil” status, transposable elements appear to be active and show high diversity, suggesting a role for them in the remarkable expansion of the lungfish genome. Analyses of single genes and gene families documented changes connected to the water to land transition and demonstrated the value of the lungfish reference transcriptome for comparative studies of vertebrate evolution. KW - lungfish KW - transcriptome KW - genome KW - sarcopterygian fish Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-167753 VL - 6 IS - 21571 ER - TY - JOUR A1 - Bluemel, Christina A1 - Linke, Fraenze A1 - Herrmann, Ken A1 - Simunovic, Iva A1 - Eiber, Matthias A1 - Kestler, Christian A1 - Buck, Andreas K. A1 - Schirbel, Andreas A1 - Bley, Thorsten A. A1 - Wester, Hans-Juergen A1 - Vergho, Daniel A1 - Becker, Axel T1 - Impact of \(^{68}\)Ga-PSMA PET/CT on salvage radiotherapy planning in patients with prostate cancer and persisting PSA values or biochemical relapse after prostatectomy JF - EJNMMI Research N2 - Background Salvage radiotherapy (SRT) is clinically established in prostate cancer (PC) patients with PSA persistence or biochemical relapse (BCR) after prior radical surgery. PET/CT imaging prior to SRT may be performed to localize disease recurrence. The recently introduced \(^{68}\)Ga-PSMA outperforms other PET tracers for detection of recurrence and is therefore expected also to impact radiation planning. Forty-five patients with PSA persistence (16 pts) or BCR (29 pts) after prior prostatectomy, scheduled to undergo SRT of the prostate bed, underwent \(^{68}\)Ga-PSMA PET/CT. The median PSA level was 0.67 ng/ml. The impact of \(^{68}\)Ga-PSMA PET/CT on the treatment decision was assessed. Patients with oligometastatic (≤5 lesions) PC underwent radiotherapy (RT), with the extent of the RT area and dose escalation being based on PET positivity. Results Suspicious lesions were detected in 24/45 (53.3 %) patients. In 62.5 % of patients, lesions were only detected by 68Ga-PSMA PET. Treatment was changed in 19/45 (42.2 %) patients, e.g., extending SRT to metastases (9/19), administering dose escalation in patients with morphological local recurrence (6/19), or replacing SRT by systemic therapy (2/19). 38/45 (84.4 %) followed the treatment recommendation, with data on clinical follow-up being available in 21 patients treated with SRT. All but one showed biochemical response (mean PSA decline 78 ± 19 %) within a mean follow-up of 8.12 ± 5.23 months. Conclusions \(^{68}\)Ga-PSMA PET/CT impacts treatment planning in more than 40 % of patients scheduled to undergo SRT. Future prospective studies are needed to confirm this significant therapeutic impact on patients prior to SRT. KW - prostate cancer KW - salvage radiotherapy KW - PSMA KW - PET/CT KW - recurrence Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-147798 VL - 6 IS - 78 ER - TY - THES A1 - Blättner, Sebastian T1 - The role of the non-ribosomal peptide synthetase AusAB and its product phevalin in intracellular virulence of Staphylococcus aureus T1 - Die Rolle der nicht-ribosomalen Peptidsynthetase AusAB und ihres Produktes Phevalin in der intrazellulären Virulenz von Staphylococcus aureus N2 - Staphylococcus aureus is a prevalent commensal bacterium which represents one of the leading causes in health care-associated bacterial infections worldwide and can cause a variety of different diseases ranging from simple abscesses to severe and life threatening infections including pneumonia, osteomyelitis and sepsis. In recent times multi-resistant strains have emerged, causing severe problems in nosocomial as well as community-acquired (CA) infection settings, especially in the United States (USA). Therefore S. aureus has been termed as a superbug by the WHO, underlining the severe health risk originating from it. Today, infections in the USA are dominated by S. aureus genotypes which are classified as USA300 and USA400, respectively. Strains of genotype USA300 are responsible for about 70% of the CA infections. The molecular mechanisms which render S. aureus such an effective pathogen are still not understood in its entirety. For decades S. aureus was thought to be a strictly extracellular pathogen relying on pore-forming toxins like α-hemolysin to damage human cells and tissue. Only recently it has been shown that S. aureus can enter non-professional phagocytes, using adhesins like the fibronectin-binding proteins which mediate an endocytotic uptake into the host cells. The bacteria are consequently localized to endosomes, where the degradation of enclosed bacterial cells through phagosome maturation would eventually occur. S. aureus can avoid degradation, and translocate to the cellular cytoplasm, where it can replicate. The ability to cause this so-called phagosomal escape has mainly been attributed to a family of amphiphilic peptides called phenol soluble modulins (PSMs), but as studies have shown, they are not sufficient. In this work I used a transposon mutant library in combination with automated fluorescence microscopy to screen for genes involved in the phagosomal escape process and intracellular survival of S. aureus. I thereby identified a number of genes, including a non-ribosomal peptide synthetase (NRPS). The NRPS, encoded by the genes ausA and ausB, produces two types of small peptides, phevalin and tyrvalin. Mutations in the ausAB genes lead to a drastic decrease in phagosomal escape rates in epithelial cells, which were readily restored by genetic complementation in trans as well as by supplementation of synthetic phevalin. In leukocytes, phevalin interferes with calcium fluxes and activation of neutrophils and promotes cytotoxicity of intracellular bacteria in both, macrophages and neutrophils. Further ausAB is involved in survival and virulence of the bacterium during mouse lung pneumoniae. The here presented data demonstrates the contribution of the bacterial cyclic dipeptide phevalin to S. aureus virulence and suggests, that phevalin directly acts on a host cell target to promote cytotoxicity of intracellular bacteria. N2 - Staphylococcus aureus ist ein weit verbreitetes kommensales Bakterium, welches zugleich einer der häufigsten Verursacher von Krankenhausinfektionen ist, und eine Reihe verschiedener Krankheiten, angefangen bei simplen Abszessen, bis hin zu schweren Erkrankungen wie Lungenentzündung, Osteomylitis und Sepsis verursachen kann. Das Risiko durch nosokomiale sowie epidemische S. aureus Infektionen ist in den vergangenen Jahren weiter gestiegen. Dazu beigetragen hat das Auftreten multiresistenter und hoch cytotoxischer Stämme, vor allem in den USA. Als Konsequenz hat die WHO S. aureus inzwischen als „Superbug“ tituliert und als globales Gesundheitsrisiko eingestuft. Bei CA-Infektionen dominieren die Isolate der Klassifizierung USA300 und USA400, wobei den Erstgenannten bis zu 70% aller in den USA registrierten CA-MRSA Infektionen der letzten Jahre zugesprochen werden. Lange Zeit wurde angenommen, dass S. aureus strikt extrazellulär im Infektionsbereich vorliegt und die cytotoxische Wirkung von z.B. α-Toxin für Wirtszelltod und Gewebeschädigungen verantwortlich ist. Erst vor kurzem wurde festgestellt, dass S. aureus auch durch fakultativ phagozytotische Zellen, wie Epithel- oder Endothelzellen, mittels zahlreicher Adhäsine aufgenommen wird. Die Aufnahme in die Zelle erfolgt zunächst in ein Phagoendosom, in dem die Pathogene durch antimikrobielle Mechanismen abgebaut würden. Um dies zu verhindern, verfügt S. aureus über Virulenzfaktoren, welche die endosomale Membran schädigen. Die Bakterien gelangen so in das Zellzytoplasma, wo sie sich vervielfältigen können, bevor die Wirtszelle schließlich getötet wird. Eine wichtige Funktion in diesem Vorgang konnte bereits in mehreren Studien den Phenol löslichen Modulinen (PSM) zugesprochen werden, Arbeiten unserer Gruppe deuten jedoch darauf hin, dass diese nicht alleine für den phagosomalen Ausbruch von S. aureus verantwortlich sind. In dieser Arbeit verwendete ich eine Transposon Mutantenbibliothek des S. aureus Stammes JE2 (USA300) in Verbindung mit automatisierter Fluoreszenzmikroskopie, um Gene zu identifizieren, die den phagosomalen Ausbruch von S. aureus beeinflussen. Unter den Mutanten, welche eine Minderung der Ausbruchsraten zeigten, fanden sich auch Mutanten in beiden Genen eines Operons, welches für die nicht-ribosomale Peptidsynthetase AusA/B codiert, die die beiden Dipeptide Phevalin und Tyrvalin produziert. Verminderte Ausbruchsraten konnten sowohl durch genetische Komplementation als auch mittels des Zusatzes synthetischen Phevalins wiederhergestellt werden. In Leukozyten verhindert Phevalin effizienten Calcium-Flux und die Aktivierung von Neutrophilen. Zudem fördert Phevalin die Cytotoxizität intrazellulärer Bakterien sowohl in Makrophagen, als auch Neutrophilen. Darüber hinaus konnten wir zeigen, dass die NRPS AusAB und ihre Produkte eine Rolle beim Überleben der Bakterien während einer Infektion im Tiermodell einnehmen. Die hier präsentierten Daten hinsichtlich des Einflusses von Phevalin auf Virulenz und der Interaktion zwischen Wirt und Pathogen lassen den Schluss zu, dass Phevalin direkt auf einen Wirtszellfaktor wirkt, um die Cytotoxicität intrazellulärer Bakterien zu stärken. KW - Staphylococcus aureus KW - MRSA KW - Virulenz KW - Intracellular virulence KW - Non-ribosomal peptide synthetase KW - USA300 Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-146662 ER - TY - JOUR A1 - Blättner, Sebastian A1 - Das, Sudip A1 - Paprotka, Kerstin A1 - Eilers, Ursula A1 - Krischke, Markus A1 - Kretschmer, Dorothee A1 - Remmele, Christian W. A1 - Dittrich, Marcus A1 - Müller, Tobias A1 - Schuelein-Voelk, Christina A1 - Hertlein, Tobias A1 - Mueller, Martin J. A1 - Huettel, Bruno A1 - Reinhardt, Richard A1 - Ohlsen, Knut A1 - Rudel, Thomas A1 - Fraunholz, Martin J. T1 - Staphylococcus aureus Exploits a Non-ribosomal Cyclic Dipeptide to Modulate Survival within Epithelial Cells and Phagocytes JF - PLoS Pathogens N2 - Community-acquired (CA) Staphylococcus aureus cause various diseases even in healthy individuals. Enhanced virulence of CA-strains is partly attributed to increased production of toxins such as phenol-soluble modulins (PSM). The pathogen is internalized efficiently by mammalian host cells and intracellular S. aureus has recently been shown to contribute to disease. Upon internalization, cytotoxic S. aureus strains can disrupt phagosomal membranes and kill host cells in a PSM-dependent manner. However, PSM are not sufficient for these processes. Here we screened for factors required for intracellular S. aureus virulence. We infected escape reporter host cells with strains from an established transposon mutant library and detected phagosomal escape rates using automated microscopy. We thereby, among other factors, identified a non-ribosomal peptide synthetase (NRPS) to be required for efficient phagosomal escape and intracellular survival of S. aureus as well as induction of host cell death. By genetic complementation as well as supplementation with the synthetic NRPS product, the cyclic dipeptide phevalin, wild-type phenotypes were restored. We further demonstrate that the NRPS is contributing to virulence in a mouse pneumonia model. Together, our data illustrate a hitherto unrecognized function of the S. aureus NRPS and its dipeptide product during S. aureus infection. KW - cell death KW - cytotoxicity KW - Staphylococcus aureus KW - host cells KW - neutrophils KW - macrophages KW - transposable elements KW - epithelial cells Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-180380 VL - 12 IS - 9 ER - TY - THES A1 - Bolaños-Rosales, Alejandro T1 - Low Mach Number Simulations of Convective Boundary Mixing in Classical Novae T1 - Simulationen der konvektiven Mischung in Klassischen Novae im Strömungsbereich kleiner Machzahlen N2 - Classical novae are thermonuclear explosions occurring on the surface of white dwarfs. When co-existing in a binary system with a main sequence or more evolved star, mass accretion from the companion star to the white dwarf can take place if the companion overflows its Roche lobe. The envelope of hydrogen-rich matter which builds on top of the white dwarf eventually ignites under degenerate conditions, leading to a thermonuclear runaway and an explosion in the order of 1046 erg, while leaving the white dwarf intact. Spectral analyses from the debris indicate an abundance of isotopes that are tracers of nuclear burning via the hot CNO cycle, which in turn reveal some sort of mixing between the envelope and the white dwarf underneath. The exact mechanism is still a matter of debate. The convection and deflagration in novae develop in the low Mach number regime. We used the Seven League Hydro code (SLH ), which employs numerical schemes designed to correctly simulate low Mach number flows, to perform two and three- dimensional simulations of classical novae. Based on a spherically-symmetric model created with aid of a stellar evolution code, we developed our own nova model and tested it on a variety of numerical grids and boundary conditions for validation. We focused on the evolution of temperature, density and nuclear energy generation rate at the layers between white dwarf and envelope, where most of the energy is generated, to understand the structure of the transition region, and its effect on the nuclear burning. We analyzed the resulting dredge-up efficiency stemming from the convective motions in the envelope. Our models yield similar results to the literature, but seem to depend very strongly on the numerical resolution. We followed the evolution of the nuclear species involved in the CNO cycle and concluded that the thermonuclear reactions primarily taking place are those of the cold and not the hot CNO cycle. The reason behind this could be that under the conditions generally assumed for multi-dimensional simulations, the envelope is in fact not degenerate. We performed initial tests for 3D simulations and realized that alternative boundary conditions are needed. N2 - Klassische Novae sind thermonukleare Explosionen an der Oberfläche von Weißen Zwergen. Wenn ein solcher sich in einem Doppelsternsystem zusammen mit einem Hauptreihenstern oder einem späteren Stern befindet, kann Akkretion vom Begleiter zum Weißen Zwerg stattfinden, falls der Begleitstern seine Roche-Grenze überschre- itet. Die wasserstoffreiche Hülle, die sich auf der Oberfläche des Sterns bildet, zündet aufgrund des enormen Gravitationsdrucks in einer Deflagration. Aufgrund der Entar- tung des Gases führt das nukleare Brennen zu einem thermonuklearen Durchgehen (engl. runaway)und schließlich zu einer Explosion mit Energien in der Größenord- nung von 1046 erg. Der Weiße Zwerg bleibt dabei unberührt. Spektralanalysen der ausgestoßenen Gase deuten auf Isotope hin, die am heißen CNO-Zyklus beteiligt sind. Dies legt nahe, dass vor oder während der Brennphase eine Durchmischung von Materie zwischen der akkretierten Hülle und dem Weißen Zwerg stattfinden muss. Die Konvektion und Deflagration entwickeln sich im Strömungsbereich kleiner Machzahlen. Wir benutzten den Seven League Hydro code (SLH ), welcher ëber numerische Verfahren verfügt, die auf einen weiten Bereich von Machzahlen anwend- bar sind. Daraus errechneten wir Simulationen von Klassischen Novae in zwei und drei Dimensionen. Basierend auf einem sphärisch-symmetrischen Modell, das wir mit einem Sternentwicklungscode erstellten, entwickelten wir ein eigenes Nova-Modell. Wir testeten dies in Kombination mit eienr Reihe von Gittern und Randbedingun- gen. Anschließend analysierten wir im Detail das Verhalten von Temperatur, Dichte und nuklearer Energieerzeugungsrate in den Schichten zwischen Weißem Zwerg und Wasserstoffhülle, wo die Kernfusion hauptsächlich stattfindet, um die Struktur der Brennzone und deren Einfluss auf die Nukleosynthese zu verstehen. Wir analysierten die Effizienz der Konvektion, welche Elemente aus dem Weißen Zwerg nach oben in die Hülle transportiert. Die Ergebnisse entsprechen denen der Literatur, dennoch hängen sie stark von der numerischen Auflösung ab. Wir untersuchten die Isotopen- häufigkeit der im CNO-Zyklus beteiligten Elemente, und schloßen hieraus, dass das Brennen durch den weniger energetischen “kalten” CNO-Zyklus verläuft. Dies kann darauf zurückgeführt werden, dass unter den Bedingungen, die die Mehrzahl der multi- dimensionalen Modelle aus der Fachliteratur mit sich bringen, die Wasserstoffhülle tatsächlich nicht entartet ist. Abschließend simulierten wir testweise 3D-Modelle, aber neue Randbedingungen sind nötig, um mit den Berechnungen fortfahren zu können. KW - Nova KW - Weißer Zwerg KW - classical novae KW - convection KW - numerical hydrodynamics KW - nuclear reactions KW - stellar evolution KW - simulation KW - low Mach number flows KW - numerische Hydrodynamik KW - thermonukelare Reaktionen KW - Sternentwicklung Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-132863 ER - TY - JOUR A1 - Boll, Sabine A1 - Bartholomaeus, Marie A1 - Peter, Ulrike A1 - Lupke, Ulrike A1 - Gamer, Matthias T1 - Attentional mechanisms of social perception are biased in social phobia JF - Journal of Anxiety Disorders N2 - Previous studies of social phobia have reported an increased vigilance to social threat cues but also an avoidance of socially relevant stimuli such as eye gaze. The primary aim of this study was to examine attentional mechanisms relevant for perceiving social cues by means of abnormalities in scanning of facial features in patients with social phobia. In two novel experimental paradigms, patients with social phobia and healthy controls matched on age, gender and education were compared regarding their gazing behavior towards facial cues. The first experiment was an emotion classification paradigm which allowed for differentiating reflexive attentional shifts from sustained attention towards diagnostically relevant facial features. In the second experiment, attentional orienting by gaze direction was assessed in a gaze-cueing paradigm in which non-predictive gaze cues shifted attention towards or away from subsequently presented targets. We found that patients as compared to controls reflexively oriented their attention more frequently towards the eyes of emotional faces in the emotion classification paradigm. This initial hypervigilance for the eye region was observed at very early attentional stages when faces were presented for 150 ms, and persisted when facial stimuli were shown for 3 s. Moreover, a delayed attentional orienting into the direction of eye gaze was observed in individuals with social phobia suggesting a differential time course of eye gaze processing in patients and controls. Our findings suggest that basic mechanisms of early attentional exploration of social cues are biased in social phobia and might contribute to the development and maintenance of the disorder. KW - Social anxiety KW - Eye gaze KW - Vigilance KW - Amygdala KW - Emotional expression KW - Gaze perception Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-189046 VL - 40 ER - TY - JOUR A1 - Born, Dennis-Peter A1 - Zinner, Christoph A1 - Düking, Peter A1 - Sperlich, Billy T1 - Multi-Directional Sprint Training Improves Change-Of-Direction Speed and Reactive Agility in Young Highly Trained Soccer Players JF - Journal of Sports Science and Medicine N2 - The aim of this study was to evaluate the effect of a repeated sprint training with multi-directional change-of-direction (COD) movements (RSmulti) compared to repeated shuttle sprints (RSS) on variables related to COD speed and reactive agility. Nineteen highly-trained male U15 soccer players were assigned into two groups performing either RSmulti or RSS. For both groups, each training session involved 20 repeated 15 s sprints interspersed with 30 s recovery. With RSmulti the COD movements were randomized and performed in response to a visual stimulus, while the RSS involved predefined 180° COD movements. Before and following the six training sessions, performance in the Illinois agility test (IAT), COD speed in response to a visual stimulus, 20 m linear sprint time and vertical jumping height were assessed. Both groups improved their performance in the IAT (p < 0.01, ES = 1.13; p = 0.01, ES = 0.55). The COD speed in response to a visual stimulus improved with the RSmulti (p < 0.01, ES = 1.03), but not the RSS (p = 0.46, ES = 0.28). No differences were found for 20 m sprint time (P=0.73, ES = 0.07; p = 0.14, ES = 0.28) or vertical jumping height (p = 0.46, ES = 0.11; p = 0.29, ES = 0.12) for the RSmulti and RSS, respectively. In conclusion, performance in the IAT improved with the RSmulti as well as RSS. With the RSmulti however, the COD movements are performed in response to a visual stimulus, which may result in specific adaptations that improve COD speed and reactive agility in young highly trained soccer players. KW - team sport KW - COD movements KW - repeated shuttle sprints KW - speed KW - Speedcourt Y1 - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-146866 UR - http://www.jssm.org/researchjssm-15-314.xml.xml VL - 15 ER -