21,685 research outputs found

    Identifizierung prÀdiktiver und prognostischer Biomarker in unterschiedlichen Tumorkompartimenten des ösophagealen Adenokarzinoms

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    Das ösophageale Adenokarzinom zeigt eine global steigende Inzidenz und hat mit einer 5-Jahres-Überlebensrate von weniger als 25% eine schlechte Prognose. Personalisierte TherapieansĂ€tze sind selten und prognostische/prĂ€diktive Biomarker des Tumormikromilieus sind unzureichend charakterisiert. Die kumulative Promotion nĂ€hert sich dieser Problematik in drei unterschiedlichen Schwerpunkten. 1. Zur Identifizierung Kompartiment-spezifischer Biomarker wurde eine Methode entwickelt, welche als kostengĂŒnstige Alternative zum sc-Seq Expressionsprofile individueller Zelltypen generiert. Dabei erfolgt die Extraktion der RNA nicht aus Einzelzellen, sondern aus flowzytometrisch-getrennten Zellkompartimenten. Die Separation der Proben in Epithelzellen, Immunzellen und Fibroblasten wurde durch verschiedene Verfahren validiert und eine suffiziente Ausbeute an RNA auch fĂŒr kleine Gewebemengen gezeigt. 2. Biomarker des Immunzellkompartiments als therapeutische Angriffspunkte wurden in einem Patientenkollektiv von bis zu 551 Patienten auf ihre Bedeutung beim EAC ĂŒberprĂŒft. Es zeigte sich eine Expression der Immuncheckpoints LAG3, VISTA und IDO auf TILs durch IHC und RNA-Sonden basierte Verfahren in einem relevanten Anteil (LAG3: 11,4%, VISTA: 29%, IDO: 52,6%). Es konnte eine prognostisch gĂŒnstige Bedeutung der VISTA, LAG3 und IDO Expression gezeigt werden. Durch den Vergleich von Genexpressionsprofilen aus therapienaiven und vorbehandelten Tumoren konnte zudem ein immunsuppressiver Effekt von neoadjuvanten Therapiekonzepten auf das Tumormikromilieu des EACs gezeigt werden. Dabei kam es zur verminderten Expression von Checkpoints und Anzahl TILs nach (Radio-) Chemotherapie. 3. Im Tumorzellkompartiment wurde die Rolle von Amplifikationen in ErbB-Rezeptor abhĂ€ngigen Signalwegen durch FISH-Technik und Immunhistochemie evaluiert. Es fanden sich KRAS Amplifikationen in 17,1%, PIK3CA Amplifikationen in 5% sowie eine HER2/neu-Überexpression in 14,9% der untersuchten Tumore

    Reinforcing optimization enabled interactive approach for liver tumor extraction in computed tomography images

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    Detecting liver abnormalities is a difficult task in radiation planning and treatment. The modern development integrates medical imaging into computer techniques. This advancement has monumental effect on how medical images are interpreted and analyzed. In many circumstances, manual segmentation of liver from computerized tomography (CT) imaging is imperative, and cannot provide satisfactory results. However, there are some difficulties in segmenting the liver due to its uneven shape, fuzzy boundary and complicated structure. This leads to necessity of enabling optimization in interactive segmentation approach. The main objective of reinforcing optimization is to search the optimal threshold and reduce the chance of falling into local optimum with survival of the fittest (SOF) technique. The proposed methodology makes use of pre-processing stage and reinforcing meta heuristics optimization based fuzzy c-means (FCM) for obtaining detailed information about the image. This information gives the optimal threshold value that is used for segmenting the region of interest with minimum user input. Suspicious areas are recognized from the segmented output. Both public and simulated dataset have been taken for experimental purposes. To validate the effectiveness of the proposed strategy, performance criteria such as dice coefficient, mode and user interaction level are taken and compared with state-of-the-art algorithms

    Therapeutic effect of mesenchymal stem cell derived extracellular vesicles on 3D model of oligocortical spheroids

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    Extracellular vesicles (EVs) are released by nearly every cell type and are an important structure in inter-cellular communication. Abnormal EV signaling is found in many conditions including ischemia, Alzheimer’s Disease (AD) and Down Syndrome (DS). However, EVs from stem cells from healthy animals have recently emerged as a possible therapeutic intervention to address a variety of neurological conditions. Mesenchymal stromal cell-derived (MSCs) extracellular vesicles from the bone marrow of young healthy monkeys contain microRNAs and proteins and previous studies have shown that MSC-EV treatment mitigates inflammation and oxidative stress, promotes myelination, and improves functional recovery in a rhesus monkey model of cortical injury. EVs have also been shown to reduce AD pathology in mouse models by promoting anti-inflammatory processes and slowing the progression of AD. While AD currently effects over 6 million people in the United States, individuals with DS are disproportionately affected by early onset AD. Therefore, investigating the efficacy of MSC-EVs as a potential therapeutic to mitigate AD like pathology in DS is critical. Accordingly, the current study aims to explore the application of EVs on 3D human brain models of DS, ischemia, and oxygen glucose deprivation (OGD). We generated human oligocortical spheroids (OLS) containing neurons, astrocytes and oligodendrocytes allowing investigation of the effects of the EVs in human, physiologically relevant conditions. First, with OLS in ischemic conditions results were insufficient in demonstrating the recapitulation of cell death and oxidative damage associated with ischemia in vivo. Consequently, the inconsistency of the model prevented us from comprehensive evaluation of the therapeutic potential of EVs in OGD model in OLS. However, we next used DS-derived OLS generated from isogenic induced pluripotent stem cell (iPSCs) lines to evaluate the efficacy of EV treatment in DS. Trisomic OLS display significantly higher levels of amyloid beta (AÎČ40 and AÎČ42) depositions in both the soluble and insoluble fractions. Additionally, trisomic OLS are consistently smaller than their euploid counterparts, and have elevated levels of cleaved-caspase 3 (CC3) detection indicating more cell death. When treated with EVs, trisomic OLS demonstrated greater preserved cortical volume, significantly decreased levels of AÎČ40 and AÎČ42 in both fractions, and significant reduction in cell death compared to the untreated trisomic OLS. These results suggest that EVs alleviated the AD-related pathology in DS-derived OLS. Evaluation of the markers of cortical layer neurons demonstrated significantly higher counts of neurons expressing deep and superficial layer markers, suggesting that EVs contributed to greater preserved cortical volume of trisomic OLS by promoting neurogenesis and alleviating trisomy-induced deficits. Our studies show for the first time the efficacy of MSC-EVs in mitigating DS and AD-related cellular phenotypes and pathological depositions in human OLS. Furthermore, oligocortical spheroids present a unique tool for a target validation of potential therapeutics

    Elite perceptions of the Victorian and Edwardian past in inter-war England

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    It is often argued by historians that members of the cultivated Elite after 1918 rejected the pre-war past. or at least subjected it to severe denigration. This thesis sets out to challenge such a view. Above all, it argues that inter-war critics of the Victorian and Edwardian past were unable to reject it even if that was what they felt inclined to do. This was because they were tied to those periods by the affective links of memory, family, and the continually unfolding consequences of the past in the present. Even the severest critics of the pre-war world, such as Lytton Strachey, were less frequently dismissive of history than ambivalent towards it. This ambivalence, it is argued, helped to keep the past alive and often to humanise it. The thesis also explores more positive estimation of Victorian and Edwardian history between the wars. It examines nostalgia for the past, as well as instances of continuity of practice and attitude. It explores the way in which inter-war society drew upon aspects of Victorian and Edwardian history both as illuminating parallels to contemporary affairs and to understand directly why the present was shaped as it was. Again, this testifies to the enduring power of the past after 1918. There are three parts to this thesis. Part One outlines the cultural context in which writers contemplated the Victorian and Edwardian past. Part Two explores some of the ways in which history was written about and used by inter-war society. Part Three examines the ways in which biographical depictions of eminent Victorians after 1918 encouraged emotional negotiation with the pas

    Beyond the icon: Core cognition and the bounds of perception

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    This paper refines a controversial proposal: that core systems belong to a perceptual kind, marked out by the format of its representational outputs. Following Susan Carey, this proposal has been understood in terms of core representations having an iconic format, like certain paradigmatically perceptual outputs. I argue that they don’t, but suggest that the proposal may be better formulated in terms of a broader analogue format type. Formulated in this way, the proposal accommodates the existence of genuine icons in perception, and avoids otherwise troubling objections

    Towards a sociology of conspiracy theories: An investigation into conspiratorial thinking on Dönmes

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    This thesis investigates the social and political significance of conspiracy theories, which has been an academically neglected topic despite its historical relevance. The academic literature focuses on the methodology, social significance and political impacts of these theories in a secluded manner and lacks empirical analyses. In response, this research provides a comprehensive theoretical framework for conspiracy theories by considering their methodology, political impacts and social significance in the light of empirical data. Theoretically, the thesis uses Adorno's semi-erudition theory along with Girardian approach. It proposes that conspiracy theories are methodologically semi-erudite narratives, i.e. they are biased in favour of a belief and use reason only to prove it. It suggests that conspiracy theories appear in times of power vacuum and provide semi-erudite cognitive maps that relieve alienation and ontological insecurities of people and groups. In so doing, they enforce social control over their audience due to their essentialist, closed-to-interpretation narratives. In order to verify the theory, the study analyses empirically the social and political significance of conspiracy theories about the Dönme community in Turkey. The analysis comprises interviews with conspiracy theorists, conspiracy theory readers and political parties, alongside a frame analysis of the popular conspiracy theory books on Dönmes. These confirm the theoretical framework by showing that the conspiracy theories are fed by the ontological insecurities of Turkish society. Hence, conspiracy theorists, most readers and some political parties respond to their own ontological insecurities and political frustrations through scapegoating Dönmes. Consequently, this work shows that conspiracy theories are important symptoms of society, which, while relieving ontological insecurities, do not provide politically prolific narratives

    RNA pull-down-confocal nanoscanning (RP-CONA), a novel method for studying RNA/protein interactions in cell extracts that detected potential drugs for Parkinson’s disease targeting RNA/HuR complexes

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    MicroRNAs (miRNAs, miRs) are a class of small non-coding RNAs that regulate gene expression through specific base-pair targeting. The functional mature miRNAs usually undergo a two-step cleavage from primary miRNAs (pri-miRs), then precursor miRNAs (pre-miRs). The biogenesis of miRNAs is tightly controlled by different RNA-binding proteins (RBPs). The dysregulation of miRNAs is closely related to a plethora of diseases. Targeting miRNA biogenesis is becoming a promising therapeutic strategy. HuR and MSI2 are both RBPs. MiR-7 is post-transcriptionally inhibited by the HuR/MSI2 complex, through a direct interaction between HuR and the conserved terminal loop (CTL) of pri-miR-7-1. Small molecules dissociating pri-miR-7/HuR interaction may induce miR-7 production. Importantly, the miR-7 levels are negatively correlated with Parkinson’s disease (PD). PD is a common, incurable neurodegenerative disease causing serious motor deficits. A hallmark of PD is the presence of Lewy bodies in the human brain, which are inclusion bodies mainly composed of an aberrantly aggregated protein named α-synuclein (α-syn). Decreasing α-syn levels or preventing α-syn aggregation are under investigation as PD treatments. Notably, α-syn is negatively regulated by several miRNAs, including miR-7, miR-153, miR-133b and others. One hypothesis is that elevating these miRNA levels can inhibit α-syn expression and ameliorate PD pathologies. In this project, we identified miR-7 as the most effective α-syn inhibitor, among the miRNAs that are downregulated in PD, and with α-syn targeting potentials. We also observed potential post-transcriptional inhibition on miR-153 biogenesis in neuroblastoma, which may help to uncover novel therapeutic targets towards PD. To identify miR-7 inducers that benefit PD treatment by repressing α-syn expression, we developed a novel technique RNA Pull-down Confocal Nanoscaning (RP-CONA) to monitor the binding events between pri-miR-7 and HuR. By attaching FITC-pri-miR-7-1-CTL-biotin to streptavidin-coated agarose beads and incubating them in human cultured cell lysates containing overexpressed mCherry-HuR, the bound RNA and protein can be visualised as quantifiable fluorescent rings in corresponding channels in a confocal high-content image system. A pri-miR-7/HuR inhibitor can decrease the relative mCherry/FITC intensity ratio in RP-CONA. With this technique, we performed several small-scale screenings and identified that a bioflavonoid, quercetin can largely dissociate the pri-miR-7/HuR interaction. Further studies proved that quercetin was an effective miR-7 inducer as well as α-syn inhibitor in HeLa cells. To understand the mechanism of quercetin mediated α-syn inhibition, we tested the effects of quercetin treatment with miR-7-1 and HuR knockout HeLa cells. We found that HuR was essential in this pathway, while miR-7 hardly contributed to the α-syn inhibition. HuR can directly bind an AU-rich element (ARE) at the 3’ untranslated region (3’-UTR) of α-syn mRNA and promote translation. We believe quercetin mainly disrupts the ARE/HuR interaction and disables the HuR-induced α-syn expression. In conclusion, we developed and optimised RP-CONA, an on-bead, lysate-based technique detecting RNA/protein interactions, as well as identifying RNA/protein modulators. With RP-CONA, we found quercetin inducing miR-7 biogenesis, and inhibiting α-syn expression. With these beneficial effects, quercetin has great potential to be applied in the clinic of PD treatment. Finally, RP-CONA can be used in many other RNA/protein interactions studies