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Dissertationen zum Thema „Real-time cell analysis“
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Hong, Soonjin Barbee Kenneth A. „Quantitative analysis of cell-surface interactions and cell adhesion process in real-time /“. Philadelphia, Pa. : Drexel University, 2008. http://hdl.handle.net/1860/2840.
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Schulz, Craig. „Microsequential injection systems for the real-time monitoring of glucose metabolism of live cells by enzymatic assay /“. Thesis, Connect to this title online; UW restricted, 2002. http://hdl.handle.net/1773/8694.
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Maiuri, Paolo. „Single-cell and real-time analysis of transcription rates from integrated HIV-1 provirus“. Doctoral thesis, Scuola Normale Superiore, 2009. http://hdl.handle.net/11384/85935.
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Viral RNA biogenesis is a crucial step in the replication of retroviruses that require both the production of a genomic RNA as well as of translation templates. Cellular and viral factors concur in the biogenesis of RNA at the specific sub-nuclear chromatin site where the reaction takes place. The possibility of tracking viral RNA in living cells gives the unique possibility of measuring the kinetic parameters of RNA biogenesis as well as defining the dynamic recruitment of host and viral factors to the site of replication. In order to study the activation of HIV-1 gene expression from the integrated viral promoter we exploited a method that allows the visualization of newly transcribed RNA in living cells through the specific recognition of an RNA consensus sequence for the bacteriophage MS2 coat protein tagged with an autofluorescent protein. We observed that transcription of HIV-1 occurred in discrete foci within the nucleus of cells carrying several tandem arrays of the HIV-1 construct. These foci, representing newly transcribed RNA, co-localized with the viral Tat transactivator as well as members of the positive transcription elongation factor (P-TEFb) and RNA polymerase II (RNAPII). This experimental setting was used to measure the dynamic of HIV-1 RNA transcription in living cells. By fluorescence recovery after photobleaching (FRAP) we were able to demonstrate that following photobleaching the process reaches a steady state with a negligible immobile fraction allowing precise kinetic measurements of RNA polymerase elongation rates. We found that elongation proceeded at approximately 2 kb/min, and that 3'-end formation and release took another minute to complete. In addition we also analyzed the dynamic of RNAPII and the TAR:Tat:pTEFb complex at the site of HIV-1 transcription in living cells. Our data suggest that, while the residence time of RNAPII exceeds the time required for elongation through the viral template, the complex dissociates from the polymerase following transcription initiation, and may undergo subsequent cycles of association/dissociation. This approach was extended to the analysis of single integrated HIV- 1 transcription units by transduction of a HIV-based lentiviral vector into a human cell line and subsequent selection for Tat-induction from a latent state. Nascent RNAs from single integrated transcription units were detectable in living cells by MS2 RNA-tagging. At steady state a constant number of RNAs was measured at the transcription site corresponding to a minimal density of polymerases with negligible fluctuations over time both in space and intensity of the signal. Recovery of fluorescence after photobleaching of the transcription site was complete within seconds, much faster than what was observed previously. However, the necessity of taking into account also the diffusion of the tagged MS2 protein required the development of novel analytical tools. To this end we developed a model that describes each polymerase sliding along the DNA like the peak of a positive progressive traveling wave (TranWave) to predict the number of MS2 RNA repeats at the transcription site in function of time. The outcome of this approach and its following improvements are being discussed. This work provides for the first time a kinetic framework to analyze HIV-1 RNA biogenesis and RNA/protein dynamics in living cells.
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Shoshi, Astrit [Verfasser]. „Magnetic lab-on-a-chip for cell analysis : magnetoresistive-based real-time monitoring of dynamic cell-environment interactions / Astrit Shoshi“. Bielefeld : Universitaetsbibliothek Bielefeld, 2013. http://d-nb.info/1036974502/34.
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Li, Min. „Kinetic analysis of Human T-cell leukemia virus type 1 gene expression“. The Ohio State University, 2008. http://rave.ohiolink.edu/etdc/view?acc_num=osu1228156327.
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Zhou, Daming. „Modeling and Multi-Dimensional Analysis of a Proton Exchange Membrane Fuel Cell“. Thesis, Bourgogne Franche-Comté, 2017. http://www.theses.fr/2017UBFCA011/document.
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Un des freins à la commercialisation de masse de la pile à combustible et notamment de la technologie à membrane échangeuse de proton vient de sa faible durée de vie due à la difficulté de contrôler le système sous certaines conditions. Pour pallier à ce problème, l’élaboration d’un modèle mathématique précis de la pile à combustible à membrane échangeuse de protons permettant d’observer les variables internes et l'état de la pile à combustible au cours de son fonctionnement permettrait le développement de la stratégie de contrôle du système.Cette thèse propose d’élaborer un modèle dynamique multi-physique complet pour la pile à combustible à membrane échangeuse de protons. Le modèle proposé couvre les domaines multi-physiques pour les caractéristiques électriques, fluidiques et thermiques. Dans ces deux derniers domaines, les phénomènes transitoires sont notamment pris en compte dans le modèle proposé, tels que les comportements dynamiques de la teneur en eau de la membrane de la pile et la température. Par conséquent, ce modèle peut être utilisé pour analyser les effets de couplage des variables dynamiques entre différents domaines physiques.Grace à ce modèle ainsi définit, un second modèle multi-physique bidimensionnel plus détaillé est également présenté. Le modèle proposé couvre les domaines électriques et fluidiques avec une approche de modélisation 2-D innovante. Les distributions spatiales de quantité physique dans le domaine électrique peuvent ainsi être obtenues. Par conséquent, ce modèle 2-D PEMFC peut être utilisé pour étudier les influences des paramètres de modélisation sur la prédiction de performance multidimensionnelle locale. Une étude expérimentale est effectuée pour valider le modèle 2-D proposé avec une pile commerciale PEMFC Ballard NEXA de 1,2 kW.Dans un second chapitre, une analyse des phénomènes dynamiques est réalisée en fonction du modèle dynamique multidisciplinaire développé en s’appuyant sur la méthode RGA (gain relatif) pour diverses variables d'entrée de contrôle, afin d'analyser quantitativement les effets de couplage dans différents domaines physiques. L’étude s’intéresse entre autre aux interactions de la teneur en eau et de la température de la membrane. L'analyse de couplage présentée dans cette thèse peut aider les ingénieurs à concevoir et à optimiser les stratégies de contrôle des piles à combustible, en particulier pour la gestion de l'eau et de la chaleur dans les systèmes de piles à combustible.Une deuxième analyse portant sur la sensibilité aux paramètres de l'étude est effectuée sur la base du modèle multidisciplinaire bidimensionnel développé. Ces résultats d'analyse de sensibilité globale fournissent des informations utiles pour la compréhension de la dégradation, le réglage des paramètres et la simplification du modèle des piles à combustible.Dans un troisième temps, le modèle proposé se décline dans un algorithme de résolution mathématique en temps réel basé sur un algorithme de matrice tri diagonal efficace (TDMA). Les résultats expérimentaux démontrent les possibilités pratiques du modèle 2-D proposé pour le contrôle en temps réel avancé des systèmes de pile à combustible avec un temps de calcul de la boucle de contrôle de l'ordre de la milliseconde. Le temps d'exécution du modèle peut être quadruplé par rapport aux algorithme séquentiels présent dans la littérature; garantissant ainsi des décisions et des actions de contrôle rapideBefore mass commercialization of proton exchange membrane fuel cell, the research on the design of appropriate control strategies and auxiliaries need to be done for achieving proton exchange membrane fuel cell (PEMFC) optimal working modes. An accurate mathematical PEMFC model can be used to observe the internal variables and state of fuel cell during its operation, and could further greatly help the system control strategy development.A comprehensive multi-physical dynamic model for PEMFC is developed in chapter I. The proposed model covers multi-physical domains for electric, fluidic and thermal features. Particularly, the transient phenomena in both fluidic and thermal domain are simultaneously considered in the proposed model, such as the dynamic behaviors of fuel cell membrane water content and temperature. Therefore, this model can be used to analyze the coupling effects of dynamic variables among different physical domains.Based on the developed multi-physical PEMFC model, a full two-dimensional multi-physical model is further presented. The proposed model covers electrical and fluidic domains with an innovative 2-D modeling approach. In order to accurately describe the characteristics of reactant gas convection in the channels and diffusion through the gas diffusion layer, the gas pressure drop in the serpentine pipeline is comprehensively analyzed by fully taking the geometric form of flow field into consideration, such as the reactant gas pressure drop due to the pipeline sharp and U-bends. Based on the developed 2-D fluidic domain modeling results, spatial physical quantity distributions in electrical domain can be further obtained. Therefore, this 2-D PEMFC model can be use to study the influences of modeling parameters on the local multi-dimensional performance prediction. The simulation and experimental test are then performed to validate the proposed 2-D model with a commercial Ballard NEXA 1.2 kW PEMFC stack.In chapter II, analyses of dynamic phenomena step responses are conducted based on the developed multi-physical dynamic PEMFC model using the relative gain array (RGA) method for various control input variables, in order to quantitatively analyze the coupling effects in different physical domains, such as the interactions of membrane water content and temperature. Based on the calculated values of relative gain array, the proposed model can be considered as a fuel cell MIMO system, which could be divided into two independent control sub-systems by minimizing parameter coupling effects between each other. Due to the closely coupled parameters in the proposed first control sub-system, a decoupling control method is recommended to achieve optimized control results. The coupling analysis presented in this thesis can help engineers to design and optimize the fuel cell control strategies, especially for the water and thermal management in fuel cell systems
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Czerwieniec, Gregg Allen. „Single cell analysis using bio-aerosol mass spectrometry : development and applications for the real-time detection of bio-warfare pathogens /“. For electronic version search Digital dissertations database. Restricted to UC campuses. Access is free to UC campus dissertations, 2005. http://uclibs.org/PID/11984.
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AL, DOSSARY REEM. „Activation of human endogenous retrovirus K and cellular modifications in human melanoma cell lines: gene expression analysis“. Doctoral thesis, Università degli Studi di Roma "Tor Vergata", 2010. http://hdl.handle.net/2108/1389.
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Presso i laboratori dell’Università di Tor Vergata è stato sviluppato un sistema in vitro dotato di caratteristiche idonee allo studio dei meccanismi che sono alla base dello sviluppo e della progressione del melanoma. Tale sistema è costituito da una linea cellulare di melanoma umano, isolata da una lesione metastatica di una paziente e caratterizzata dalla capacità di crescere in adesione (linea TVM-A12), da cui sono state ottenute due distinte linee cellulari, dotate invece della caratteristica di crescita in sospensione, derivate l’una mediante tecnica della diluizione limite (Clonesp), e l’altra riducendo la concentrazione di siero dal 10% al 1% nel terreno di coltura (linea TVM-A12sp). Il passaggio dalla fase di crescita in adesione a quella in sospensione, osservato quando le cellule TVM-A12 erano poste in condizione di ridotta concentrazione di siero (1%) era accompagnato da riduzione di espressione dell’antigene melanocitico Melan-A/MART-1 e delle molecole HLA di classe I oltre che dall’aumentata capacità di formare colonie in terreno semisolido. Il cambiamento nella modalità di crescita delle cellule risultava inoltre associata all’aumento dell’espressione di HERV-K, sia a livello di mRNA che di proteine e poteva essere inibito dal blocco dell’espressione di HERV-K, ottenuto mediante RNA interference.
Scopo del presente lavoro è stato quello di caratterizzare le linee cellulari TVM-A12, parentale e di derivazione, dotate rispettivamente di capacità di crescita in aderenza ed in sospensione, al fine di chiarire i meccanismi che sono alla base dell’acquisizione del fenotipo dotato di maggiore aggressività ed individuare il ruolo dell’attivazione di HERV-K nella progressione del melanoma. A tale scopo, è stato condotto uno studio dell’espressione genica, mediante microarray, seguito da analisi in Real-Time PCR, che ha mostrato come la transizione fenotipica delle cellule, dalla fase di crescita in adesione a quella di crescita in sospensione, risultava accompagnata dalla modulazione di numerosi geni, che è noto essere coinvolti nell’acquisizione di caratteristiche di malignità. Inoltre il profilo di espressione delle cellule a crescita in sospensione, siano esse originate per diluizione limite o per riduzione della concentrazione di siero nel mezzo di coltura, risultava essere del tutto simile. Lo studio in Real-Time, utilizzato per confermare quanto osservato mediante microarray, ha mostrato che quando le cellule di melanoma iniziavano a distaccarsi dal monostrato, in presenza di FBS all’1%, i geni BHLHB2 e MYC risultavano transitoriamente attivati. Nelle cellule in presenza di bassa concentrazione di siero e durante il passaggio verso la fase definitiva di crescita in sospensione, i geni PTEN, VEFGA, CSK, PITCH1, FOXG1A e TP53 risultavano progressivamente up-regolati. Nelle cellule di melanoma che avevano acquisito la capacità di crescere stabilmente in sospensione, si osservava infine aumento di espressione dei geni WNT3, MYCN, MYCL1, BTK, CCND2, WNT2, TIMP3, IRF3, GTF2I, CTNNB1, E2F1, ARHGAP5, ARHGEF5, GPR39 e ITGB4. Riduzione di espressione dei geni ANXA7, CTNNA1, NME1, RRM1, CDKN1A, XRCC6,
HDAC, TRAM1, CD59 e TOB1 veniva riscontrata invece nelle cellule ancora adese, in presenza di 1 % di siero ed in quelle già in sospensione, rispetto alla linea di origine mantenuta in condizioni di coltura standard (10%FBS).
In questo studio è stato per la prima volta descritto un sistema cellulare in cui è stata dimostrata la correlazione tra aumento di espressione di HERV-K e acquisizione di un fenotipo più aggressivo, nonché la modulazione dei geni che in tali processi risultano coinvolti. Tale modello può fornire pertanto un utile strumento per la comprensione dei meccanismi che regolano lo sviluppo e la progressione del melanoma, nonché per la valutazione di agenti farmacologici e modulatori di geni attivi nei confronti dell’espressione di HERV-K e nella progressione del melanoma.
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Boussemaere, Luc. „Investigating off-axis digital holographic microscopy with a source of partial spatial coherence as a real-time sensor for cell cultures“. Doctoral thesis, Universite Libre de Bruxelles, 2015. http://hdl.handle.net/2013/ULB-DIPOT:oai:dipot.ulb.ac.be:2013/209086.
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Bio-pharmaceutical industry is a vast growing market and recent recommendations of the Food and Drug Administration have put a large emphasis on the characterization of biological processes and models. As a consequence, there is a high incentive on developing modern sensors in order to more accurately monitor and control processes. In that way, Digital Holographic Microscopy (DHM) presents unique features thanks to the refocusing and quantitative phase contrast imaging capabilities. In this thesis we investigate the usage of DHM to monitor yeast cultures that are often used in both the bio-pharmaceutical and bread industries and lay the basis of a methodological framework for the study of in-line cell cultures in the context of process control. We begin with a description of Digital Holography and the microscopy setup used in the thesis as well as a detailed explanation of the image processing required to extract the holographic data and its implementation on GPU with some speed execution figures given for three popular programming paradigms. We then describe the flow setup used and infer the limitations on the dynamic range of the technique due to both Poisson statistics and overlapping phenomena. Finally, we describe an algorithm that extracts the cells position, count and morphological information such as the size, aspect ratio, circularity and refraction index. Some experimental results are presented for yeasts before drawing a general overview of the technology and its dependencies. We further end with some conclusions concerning the technology and a brief comparison with existing competitors.Doctorat en Sciences de l'ingénieur
info:eu-repo/semantics/nonPublished
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Pretorius, Ashley. „Functional analysis of the mouse RBBP6 gene using Interference RNA“. Thesis, University of the Western Cape, 2007. http://etd.uwc.ac.za/index.php?module=etd&action=viewtitle&id=gen8Srv25Nme4_4435_1264363734.
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The aim of this thesis was to investigate the cellular role of the mouse RBBP6 gene using the interference RNA (RNAi) gene targeting technology and also to understand the relevance of two promoters for the RBBP6 gene.
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Thompson, Bruce Thomas. „Fourier transform infrared spectroscopic measurement of carbon monoxide and nitric oxide in sidestream cigarette smoke in real time using a hollow waveguide gas cell and nonimaging optics“. Diss., Available online, Georgia Institute of Technology, 2004:, 2004. http://etd.gatech.edu/theses/available/etd-06232004-172923/unrestricted/thompson%5Fbruce%5Ft%5F200407%5Fphd.pdf.
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Thesis (Ph. D.)--Chemistry and Biochemistry, Georgia Institute of Technology, 2005. Directed by Boris Mizaikoff.Hunt, William, Committee Member ; Weck, Marcus, Committee Member ; Mizaikoff, Boris, Committee Chair ; Janata, Jiri, Committee Member ; Orlando, Thomas, Committee Member. Includes bibliographical references.
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Du, Toit Jean. „An analysis of the effect of transformation on global– and gene–specific DNA methylation in four cultured cell lines / Jean du Toit“. Thesis, North-West University, 2010. http://hdl.handle.net/10394/4560.
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DNA methylation plays a role in several biological functions, such as gene expression regulation, and several endogenous and exogenous factors affect these DNA methylation patterns in the cell. One such alteration of a cell line's DNA methylation pattern is caused by the insertion of a vector into the cell line. Using the cytosine–extension assay and realtime methylation–specific PCR, alterations of DNA methylation levels on both global and gene–specific levels were investigated. In some cell lines the cellular transformation led to an increase in DNA methylation levels, and in others a decrease in DNA methylation amounts was observed. The same phenomenon was seen in the promoter regions of specific genes, showing that vector–insertion into a cell line caused DNA methylation alterations in many regions of the genome. These alterations in DNA methylation are investigated in this reduced representation study using enrichment of the methylated fraction of fragmented DNA and subsequent GS FLX Titanium sequencing of these methylated fragments. The results of sequence data analysis showed that methylated fragments are distributed over the whole genome, but could be related to only a few specific genes. These results have implications for cell culture work, biotechnological applications and uses in gene therapy.Thesis (M.Sc. (Biochemistry))--North-West University, Potchefstroom Campus, 2011.
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Normand, Camille. „La rhinοpneumοnie équine : caractérisatiοn mοléculaire et cellulaire de l'herpèsvirus équin 4, un mοdèle d'étude pοur l'appοrt de cοnnaissances dans la pathοgénie de la maladie, la survie et l'intégrité du virus ainsi que l'identificatiοn de mοlécules antivirales“. Electronic Thesis or Diss., Normandie, 2024. http://www.theses.fr/2024NORMC405.
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Jusqu’en 1981, l’herpèsvirus équin 4 (HVE-4) et l’HVE-1, responsables de la rhinopneumonie, étaient considérés comme deux sous-types d’un même virus. Il a depuis été démontré que l’HVE-4 était très peu impliqué dans les avortements et son implication dans la forme nerveuse de la maladie n’est pas démontrée à ce jour. C’est probablement la raison qui fait que ce virus est moins étudié que l’HVE-1. Ce travail de thèse a porté sur trois aspects importants de la rhinopneumonie chez le cheval afin d’apporter de la connaissance sur l’HVE-4 et d’évaluer l’intérêt d’utiliser ce virus comme modèle. Nous avons pu démontrer en comparant deux épizooties dans deux haras au profil vaccinal différents et en mesurant l’excrétion d’HVE-4 et la virémie par qPCR, l’apparition de séroconversions, l’intérêt de la vaccination. Si la contamination se fait essentiellement par contact, l’impact de la survie du virus dans l’environnement doit être étudiée. Nous avons montré que l’HVE-4 pouvait survivre au moins 28 jours à 4°C dans l’eau mais également sur différentes surfaces. Nous avons aussi développé une méthode d’integrity-PCR pour différencier les virus infectieux des virus non infectieux. Enfin, nous avons criblé un nombre de molécules importants par RTCA et démontré l’efficacité de plusieurs d’entre elles dont la décitabine pour laquelle nous avons réalisé une étude préliminaire du mode d’action par une analyse transcriptomiqueUntil 1981, equine herpesvirus 4 (EHV-4) and EHV-1, which cause rhinopneumonitis, were considered to be two subtypes of the same virus. Since then, it has been shown that EHV-4 is only marginally involved in abortions, and its involvement in the nervous form of the disease has not yet been demonstrated. This is probably why this virus is less studied than HVE-1. This thesis focused on three important aspects of rhinopneumonitis in horses in order to gain a better understanding of EHV 4 and to assess the value of using this virus as a model. By comparing two epizootics at two stud farms with different vaccination profiles and measuring EHV-4 excretion and viremia by qPCR, we were able to demonstrate the benefits of vaccination and the appearance of seroconversions. Although contamination occurs mainly through contact, the impact of the virus's survival in the environment needs to be studied. We have shown that EHV-4 can survive for at least 28 days at 4 °C in water and on various surfaces. We have also developed an integrity-PCR method to differentiate between infectious and non-infectious viruses. Finally, we have screened a number of compounds using RTCA and demonstrated the efficacy of several of them, including decitabine, for which we have carried out a preliminary study of the mode of action using transcriptomic analysis
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Abrahams, Beynon. „The effects of various combinations of different classes of anticancer drugs and tyrosine kinase inhibitors on the human MCF-7 breast carcinoma cell line“. Thesis, University of the Western Cape, 2014. http://hdl.handle.net/11394/3846.
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Magister Scientiae (Medical Bioscience) - MSc(MBS)This study investigated the effects of TKIs on the growth and proliferation of MCF-7 breast carcinoma cells in culture. MCF-7 cells were exposed to different concentrations of TKIs alone and in combination with each other. Inhibition of cell growth by TKIs used individually occurred in a dose- and time-dependent manner. When EGFR Inhibitor I, EGFR Inhibitor II/BIBX1382 and the multi-specific EGFR/ErbB-2/ErB-4 Inhibitor were used in combination with each other at equimolar log dose concentrations, the combined effects on cell growth was significantly different to inhibitors used individually as reflected in a decreased EC50 (IC50) during combination treatments. Generally, for the combinations with DOX, CPL and the TKIs, synergistic as well as antagonistic effects were observed at isoeffective concentrations with resultant decreases in dose reduction indices (DRIs) implying greater efficacies with the respective combinations. In this study, conventional PCR was used to detect and illustrate the presence of the EGFR gene in the samples, while RT-qPCR was used to determine the mRNA expression levels of this gene in MCF-7 breast carcinoma cells
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Xavier, Flavia Dias. „Padrão de expressão e significado prognóstico dos genes BCL2, BCL6, CCND2, FN1, LMO2 e SCYA3 pela técnica de PCR em tempo real com linfoma difuso de grandes células B tratado com rituximabe“. Universidade de São Paulo, 2013. http://www.teses.usp.br/teses/disponiveis/5/5167/tde-24062013-114437/.
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Introdução: O linfoma difuso de grandes células B é o mais freqüente grupo de linfoma não- Hodgkin, perfazendo quase 50% dos casos no serviço de hematologia do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo e Instituto do Câncer do Estado de São Paulo. Possui heterogeneidade clínica e biológica traduzida em mais de vinte subtipos na Organização Mundial da Saúde. Sua terapêutica se baseia na associação do anticorpo monoclonal anti-CD20 e quimioterapia com antracíclico, esquema que resulta em 43,5% de sobrevida global em 10 anos. Determinantes de prognóstico clínico como o Índice Internacional de Prognóstico e o Índice Internacional de Prognóstico Revisado carecem de acurácia, pois até 20% dos pacientes de baixo risco falecerão da doença e 60% dos pacientes de alto risco estarão vivos em quatro anos. Essas discrepâncias podem, em parte, ser atribuídas a fatores genéticos. A assinatura gênica do linfoma difuso de grandes células B tipo centro germinativo apresenta sobrevida global superior ao tipo células B ativadas (76% versus 16%, p=0,01), contudo o perfil de expressão gênica por microarray ainda não está disponível na prática clínica. Entretanto, o escore preditivo de mortalidade para linfoma difuso de grandes células B baseado no valor prognóstico da expressão dos genes BCL2, BCL6, CCND2, FN1, LMO2 e SCYA3 por PCR em tempo real quantitativa mostrou-se independente do Índice Internacional de Prognóstico na era pré-rituximabe. Mas não foi significante em pacientes de alto risco clínico tratados com R-CHOP. Os genes BCL2, CCND2 e SCYA3 integram a assinatura de células B ativadas, BCL6 e LMO2 a do centro germinativo e FN1 a linfonodal. Objetivo: Avaliar o impacto da expressão absoluta dos genes BCL2, BCL6, CCND2, FN1, LMO2 e SCYA3 em população brasileira com linfoma difuso de grandes células B tratada com R-CHOP em relação à resposta global, sobrevida livre de doença, sobrevida livre de progressão e sobrevida global. Métodos: A expressão gênica foi analisada por PCR em tempo real quantitativa de RNA extraído de amostras parafinadas de 63 pacientes, porém foi avaliável em 42. Seus valores foram normatizados pelo gene endógeno ABL e transformados em escala logarítmica na base 2 para posterior correlação com variáveis clínicas e de desfecho. Resultados: Com mediana de seguimento de 29 meses, as sobrevidas global, livre de doença e livre de progressão foram, respectivamente, 82,8%, 97,14% e 87,53%, enquanto a resposta completa foi 82,5%. A expressão de LMO2>3logs e BCL6>3,5logs definiu um grupo de maior sobrevida global (91% versus 64,3%, p=0,040) e sobrevida livre de doença (95,5% versus 70,7%, p=0,03), independentemente do Índice Internacional de Prognóstico (p=0,010 e p=0,042) e com significativa hiperexpressão do SCYA3 (p=0,046). Não se observou associação entre escore preditivo de mortalidade baseado nos seis genes e prognóstico. Assim, foi criado novo escore genético prognóstico baseado no poder da expressão concomitante de LMO2 e CCND2, definindo-se grupos de baixo risco (<2,5) e alto risco (>=2,5) com distintas sobrevidas global (92,4% versus 57,1%, p=0,011) e livre de progressão (96,2% versus 66,7%, p=0,013), independentes do IPI. Conclusão: Em pacientes com linfoma difuso de grandes células B tratados com R-CHOP, a hiperexpressão de BCL6, LMO2 e SCYA3 correlacionou-se com melhor prognóstico. O novo escore genético prognóstico definido por LMO2 e CCND2 estratificou grupos de risco de prognósticos distintos independentes do Índice Internacional de PrognósticoIntroduction: Diffuse large B-cell lymphoma is the most common type of non-Hodgkin lymphoma; which accounts for almost 50% of the cases at the Hematology Department of Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo and Instituto do Câncer do Estado de São Paulo. Its clinical and biological heterogeneity results in more than twenty subtypes according to the World Health Organization classification. Its treatment is based on a combination of anti-CD20 monoclonal antibody and antracycline-based chemotherapy, with a 10-year overall survival of 43.5%. Clinical prognostic determinants such as the International Prognostic Index and the Revised International Prognostic Index lack accuracy, since up to 20% of low-risk patients will die from the disease and up to 60% of high-risk patients will be alive within four years. Such discrepancies can partially be attributed to genetic factors. Diffuse large B-cell lymphoma germinal center gene signature shows superior overall survival compared to activated B-cell signature (76% versus 16%, p=0.01), however microarray gene expression profile is not yet available in clinical practice. Nonetheless, the Mortality Predictor Score for diffuse large B-cell lymphoma based on the prognostic value of BCL2, BCL6, CCND2, FN1, LMO2 and SCYA3 gene expression by quantitative real-time PCR has proved to be independent from the International Prognostic Index in the pre-rituximab era. But it was not significant in high clinical risk patients treated with R-CHOP. The genes BCL2, CCND2 and SCYA3 compose activated B-cell signature, whereas BCL6 and LMO2 compose the germinal center signature and FN1 the lymph-node signature. Objective: Evaluate the impact of BCL2, BCL6, CCND2, FN1, LMO2 and SCYA3 absolute gene expression in Brazilian population diagnosed with diffuse large B-cell lymphoma and treated with R-CHOP, with respect to overall response, disease free survival, progression free survival and overall survival. Methods: Gene expression was analyzed by quantitative real-time PCR of RNA extracted from paraffin-embedded samples of 63 patients, although evaluable in 42. Their values were normalized by endogenous gene ABL and log- transformed on a base 2 scale for subsequent correlation with clinical and outcome variables. Results: With a median follow-up of 29 months, overall survival, disease free survival and progression free survival accounted for 82.8%, 97.14% and 87.53% respectively, while complete response was 82.5%. The expression of LMO2>3logs and BCL6>3.5logs defined a group with higher overall survival (91% versus 64.3%, p=0.040) and progression free survival (95.5% versus 70.7%, p=0.03), independent of International Prognostic Index (p=0.010 and p=0.042) and with significant overexpression of SCYA3 (p=0.046). It was not identified any association between six gene Mortality Predictor Score and prognosis. As a result, we developed the New Genetic Prognostic Score based on the power of concomitant expression of LMO2 and CCND2, defining low-risk (<2.5) and high-risk (>=2.5) groups with distinct overall survival (92.4% versus 57.1%, p=0.011) and progression free survival (96.2% versus 66.7%, p=0.013), independent of International Prognostic Index. Conclusion: In patients with diffuse large B-cell lymphoma treated with R-CHOP, hyperexpression of BCL6, LMO2 and SCYA3 was correlated with a better prognosis. The New Genetic Prognostic Score, defined by LMO2 and CCND2, stratified risk groups with different prognosis, independent of International Prognostic Index
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Puebla-Osorio, Nahum. „Transcriptional analysis of chicken immune cells following exposure to 2,3,7,8,-tetrachlorodibenzo-p-dioxin (TCDD)“. Texas A&M University, 2004. http://hdl.handle.net/1969.1/3165.
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In the present investigation, microarray analysis was used to identify potential TCDD gene targets. Three microarray experiments were performed to study the effect of TCDD in an established chicken B-cell line (DT40), in a chicken macrophage cell line (HD11), and in the bursa of Fabricius from embryos exposed in ovo at 6 days of incubation. From the DT40 microarray analyses, clones with sequence similarity to the apoptotic genes caspase 8 and caspase 9, and the transcription factor NFΜB, among others, were identified. Real-time quantitative polymerase chain reaction (RT-PCR) revealed that TCDD elicits aryl hydrocarbon receptor (AhR)-mediated apoptosis in the avian DT40 pre-B-cell line through activation of caspases 9 and 3 (see chapter III). During the course of the HD11 microarray analyses, a consistent down-regulation of the matrix metalloprotease MMP-2 was observed. This finding was the basis for the hypothesis that TCDD has an effect on the gene expression of the MMP-2 and MMP-9 in macrophages. Then, gene expression analysis and functional zymography showed that TCDD impairs the MMP-2 and MMP-9 response to LPS stimulation in HD11 chicken macrophages (see chapter V). The microarray analyses of the embryonic bursa of Fabricius provided the basis to further study of the effect of TCDD in the chicken embryo. The shifted genes were classified according to their function. The down-regulated genes included: precursor of matrix metalloprotease-inhibitor, histone acyl-transferase 1, homeobox protein CUX-2, Death Associated Protein Kinase, and UDPglucosyl transferase, among others. The up-regulated genes included: phosphoinositidespecific phospholipase, acyl Co-A oxidase, and protein effector of Cdc42, among others.
Together, these microarray analyses produced a database of genes of interest that will provide sufficient hypotheses to inspire multiple investigations aimed at confirming and refining the gene expression alterations as a consequence of TCDD exposure.
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Giedt, Randy James. „Real-Time Acquisition and Analysis of Endothelial Mitochondrial Superoxide Radical Production and Membrane Potential During In Vitro Ischemia/Reperfusion“. The Ohio State University, 2009. http://rave.ohiolink.edu/etdc/view?acc_num=osu1243541457.
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18
Albuquerque, Andreia de, Sepp Kaul, Georg Breier, Petra Krabisch und Nikos Fersis. „Multimarker Analysis of Circulating Tumor Cells in Peripheral Blood of Metastatic Breast Cancer Patients: A Step Forward in Personalized Medicine“. Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2014. http://nbn-resolving.de/urn:nbn:de:bsz:14-qucosa-136627.
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Aim: To develop an immunomagnetic assay for the isolation of circulating tumor cells (CTCs) followed by the analysis of a multimarker panel, which will enable the characterization of these malignant cells with high accuracy. Patients and Methods: Peripheral blood (PB) was collected from 32 metastatic breast cancer patients and 42 negative controls. The antibodies BM7 and VU1D9 were used for immunomagnetic tumor cell enrichment. A real-time reverse transcription-polymerase chain reaction (RT-PCR) approach for the markers KRT19, SCGB2A2, MUC1, EPCAM, BIRC5 and ERBB2 was used for CTC detection and characterization. Results: The positivity rates for each marker were as follows: 46.9% for KRT19, 25.0% for SCGB2A2, 28.1% for MUC1, 28.1% for EPCAM, 21.9% for BIRC5, and 15.6% for ERBB2. After the creation of individualized cutoffs, the sensitivity and specificity of the combined marker gene panel increased to 56.3% and 100%, respectively. Interestingly, 27.0% of the HER2-negative tumor patients showed ERBB2 mRNA-positive CTCs. Conclusions: The described technique can be used to measure CTCs with great accuracy. The use of a multimarker panel for the characterization of CTCs may provide real-time information and be of great value in therapy monitoringZiel: Entwicklung eines immunomagnetischen Verfahrens zur Isolierung zirkulierender Tumorzellen (CTCs) in Kombination mit einer molekularen Multimarkeranalyse für die hochspezifische Identifizierung maligner Zellen. Patientinnen und Methoden: Peripheres Blut (PB) von 32 Patientinnen mit metastasiertem Mammakarzinom und von 42 gesunden Kontrollen wurde für die immunomagnetische Tumorzellanreicherung mit den Antikörpern BM7 und VU1D9 genutzt. Eine Real-Time Reverse Transkription Polymerase-Kettenreaktion (RT-PCR)-Methodik mit den Markern KRT19, SCGB2A2, MUC1, EPCAM, BIRC5 und ERBB2 wurde für den CTC-Nachweis und die Tumorzellcharakterisierung entwickelt. Ergebnisse: Für die einzelnen Marker wurden die folgenden Positivitätsraten ermittelt: 46,9% für KRT19, 25,0% für SCGB2A2, 28,1% für MUC1, 28,1% für EPCAM, 21,9% für BIRC5 und 15,6% für ERBB2. Nach der Bestimmung individualisierter Cut-off-Werte ergab sich für den kombinierten Multimarkernachweis eine Sensitivität und Spezifität von 56,3% bzw. 100%. Bemerkenswert war der Befund, dass 27,0% der HER2-tumornegativen Patientinnen ERBB2-mRNA-positive CTCs aufwiesen. Schlussfolgerung: Die hier beschriebene Methodik bestimmt CTCs mit hoher Spezifität. Die molekulare Multimarkeranalyse liefert wertvolle Real-Time-Informationen für personalisierte Behandlungsmodalitäten
Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich
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Albuquerque, Andreia de, Sepp Kaul, Georg Breier, Petra Krabisch und Nikos Fersis. „Multimarker Analysis of Circulating Tumor Cells in Peripheral Blood of Metastatic Breast Cancer Patients: A Step Forward in Personalized Medicine“. Karger, 2012. https://tud.qucosa.de/id/qucosa%3A27718.
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Aim: To develop an immunomagnetic assay for the isolation of circulating tumor cells (CTCs) followed by the analysis of a multimarker panel, which will enable the characterization of these malignant cells with high accuracy. Patients and Methods: Peripheral blood (PB) was collected from 32 metastatic breast cancer patients and 42 negative controls. The antibodies BM7 and VU1D9 were used for immunomagnetic tumor cell enrichment. A real-time reverse transcription-polymerase chain reaction (RT-PCR) approach for the markers KRT19, SCGB2A2, MUC1, EPCAM, BIRC5 and ERBB2 was used for CTC detection and characterization. Results: The positivity rates for each marker were as follows: 46.9% for KRT19, 25.0% for SCGB2A2, 28.1% for MUC1, 28.1% for EPCAM, 21.9% for BIRC5, and 15.6% for ERBB2. After the creation of individualized cutoffs, the sensitivity and specificity of the combined marker gene panel increased to 56.3% and 100%, respectively. Interestingly, 27.0% of the HER2-negative tumor patients showed ERBB2 mRNA-positive CTCs. Conclusions: The described technique can be used to measure CTCs with great accuracy. The use of a multimarker panel for the characterization of CTCs may provide real-time information and be of great value in therapy monitoring.Ziel: Entwicklung eines immunomagnetischen Verfahrens zur Isolierung zirkulierender Tumorzellen (CTCs) in Kombination mit einer molekularen Multimarkeranalyse für die hochspezifische Identifizierung maligner Zellen. Patientinnen und Methoden: Peripheres Blut (PB) von 32 Patientinnen mit metastasiertem Mammakarzinom und von 42 gesunden Kontrollen wurde für die immunomagnetische Tumorzellanreicherung mit den Antikörpern BM7 und VU1D9 genutzt. Eine Real-Time Reverse Transkription Polymerase-Kettenreaktion (RT-PCR)-Methodik mit den Markern KRT19, SCGB2A2, MUC1, EPCAM, BIRC5 und ERBB2 wurde für den CTC-Nachweis und die Tumorzellcharakterisierung entwickelt. Ergebnisse: Für die einzelnen Marker wurden die folgenden Positivitätsraten ermittelt: 46,9% für KRT19, 25,0% für SCGB2A2, 28,1% für MUC1, 28,1% für EPCAM, 21,9% für BIRC5 und 15,6% für ERBB2. Nach der Bestimmung individualisierter Cut-off-Werte ergab sich für den kombinierten Multimarkernachweis eine Sensitivität und Spezifität von 56,3% bzw. 100%. Bemerkenswert war der Befund, dass 27,0% der HER2-tumornegativen Patientinnen ERBB2-mRNA-positive CTCs aufwiesen. Schlussfolgerung: Die hier beschriebene Methodik bestimmt CTCs mit hoher Spezifität. Die molekulare Multimarkeranalyse liefert wertvolle Real-Time-Informationen für personalisierte Behandlungsmodalitäten.
Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.
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Zikmund, Tomáš. „Matematické metody pro zpracování obrazu v biologických pozorováních“. Doctoral thesis, Vysoké učení technické v Brně. Fakulta strojního inženýrství, 2014. http://www.nusl.cz/ntk/nusl-234208.
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The dissertation deals with the image processing in digital holographic microscopy and X-ray computed tomography. The focus of the work lies in the proposal of data processing techniques to meet the needs of the biological experiments. Transmitted light holographic microscopy is particularly used for quantitative phase imaging of transparent microscopic objects such as living cells. The phase images are affected by the phase aberrations that make the analysis particularly difficult. Here, we present a novel algorithm for dynamical processing of living cells phase images in a time-lapse sequence. The algorithm compensates for the deformation of a phase image using weighted least squares surface fitting. Moreover, it identifies and segments the individual cells in the phase image. This property of the algorithm is important for real-time cell quantitative phase imaging and instantaneous control of the course of the experiment. The efficiency of the propounded algorithm is demonstrated on images of rat fibrosarcoma cells using an off-axis holographic microscope. High resolution X-ray computed tomography is increasingly used technique for the study of the small rodent bones micro-structure. In this part of the work, the trabecular and cortical bone morphology is assessed in the distal half of rat femur. We developed new method for mapping the cortical position and dimensions from a central longitudinal axis with one degree angular resolution. This method was used to examine differences between experimental groups. The bone position in tomographic slices is aligned before the mapping using the propound standardization procedure. The activity of remodelling process of the long bone is studied on the system of cortical canals.
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de, Albuquerque Andreia, Ilja Kubisch, Georg Breier, Gudrun Stamminger, Nikos Fersis, Astrid Eichler, Sepp Kaul und Ulrich Stölzel. „Multimarker Gene Analysis of Circulating Tumor Cells in Pancreatic Cancer Patients: A Feasibility Study“. Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2014. http://nbn-resolving.de/urn:nbn:de:bsz:14-qucosa-133530.
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Objective: The aim of this study was to develop an immunomagnetic/real-time reverse transcriptase polymerase chain reaction (RT-PCR) assay and assess its clinical value for the molecular detection of circulating tumor cells (CTCs) in peripheral blood of pancreatic cancer patients. Methods: The presence of CTCs was evaluated in 34 pancreatic cancer patients before systemic therapy and in 40 healthy controls, through immunomagnetic enrichment, using the antibodies BM7 and VU1D9 [targeting mucin 1 and epithelial cell adhesion molecule (EpCAM), respectively], followed by real-time RT-PCR analysis of the genes KRT19, MUC1, EPCAM, CEACAM5 and BIRC5. Results: The developed assay showed high specificity, as none of the healthy controls were found to be positive for the multimarker gene panel. CTCs were detected in 47.1% of the pancreatic cancer patients before the beginning of systemic treatment. Shorter median progression-free survival (PFS) was observed for patients who had at least one detectable tumor-associated transcript, compared with patients who were CTC negative. Median PFS time was 66.0 days [95% confidence interval (CI) 44.8–87.2] for patients with baseline CTC positivity and 138.0 days (95% CI 124.1–151.9) for CTC-negative patients (p = 0.01, log-rank test). Conclusion: Our results suggest that in addition to the current prognostic methods, CTC analysis represents a potential complementary tool for prediction of outcome in pancreatic cancer patientsDieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich
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de, Albuquerque Andreia, Ilja Kubisch, Georg Breier, Gudrun Stamminger, Nikos Fersis, Astrid Eichler, Sepp Kaul und Ulrich Stölzel. „Multimarker Gene Analysis of Circulating Tumor Cells in Pancreatic Cancer Patients: A Feasibility Study“. Karger, 2012. https://tud.qucosa.de/id/qucosa%3A27513.
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Objective: The aim of this study was to develop an immunomagnetic/real-time reverse transcriptase polymerase chain reaction (RT-PCR) assay and assess its clinical value for the molecular detection of circulating tumor cells (CTCs) in peripheral blood of pancreatic cancer patients.
Methods: The presence of CTCs was evaluated in 34 pancreatic cancer patients before systemic therapy and in 40 healthy controls, through immunomagnetic enrichment, using the antibodies BM7 and VU1D9 [targeting mucin 1 and epithelial cell adhesion molecule (EpCAM), respectively], followed by real-time RT-PCR analysis of the genes KRT19, MUC1, EPCAM, CEACAM5 and BIRC5.
Results: The developed assay showed high specificity, as none of the healthy controls were found to be positive for the multimarker gene panel. CTCs were detected in 47.1% of the pancreatic cancer patients before the beginning of systemic treatment. Shorter median progression-free survival (PFS) was observed for patients who had at least one detectable tumor-associated transcript, compared with patients who were CTC negative. Median PFS time was 66.0 days [95% confidence interval (CI) 44.8–87.2] for patients with baseline CTC positivity and 138.0 days (95% CI 124.1–151.9) for CTC-negative patients (p = 0.01, log-rank test).
Conclusion: Our results suggest that in addition to the current prognostic methods, CTC analysis represents a potential complementary tool for prediction of outcome in pancreatic cancer patients.Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.
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PIZZOLATO, ALBERTO. „Topology optimization for energy problems“. Doctoral thesis, Politecnico di Torino, 2018. http://hdl.handle.net/11583/2710567.
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The optimal design of energy systems is a challenge due to the large design space and the complexity of the tightly-coupled multi-physics phenomena involved.
Standard design methods consider a reduced design space, which heavily constrains the final geometry, suppressing the emergence of design trends. On the other hand, advanced design methods are often applied to academic examples with reduced physics complexity that seldom provide guidelines for real-world applications. This dissertation offers a systematic framework for the optimal design of energy systems by coupling detailed physical analysis and topology optimization.
Contributions entail both method-related and application-oriented innovations.
The method-related advances stem from the modification of topology optimization approaches in order to make practical improvements to selected energy systems. We develop optimization models that respond to realistic design needs, analysis models that consider full physics complexity and design models that allow dramatic design changes, avoiding convergence to unsatisfactory local minima and retaining analysis stability.
The application-oriented advances comprise the identification of novel optimized geometries that largely outperform industrial solutions. A thorough analysis of these configurations gives insights into the relationship between design and physics, revealing unexplored design trends and suggesting useful guidelines for practitioners.
Three different problems along the energy chain are tackled. The first one concerns thermal storage with latent heat units. The topology of mono-scale and multi-scale conducting structures is optimized using both density-based and level-set descriptions. The system response is predicted through a transient conjugate heat transfer model that accounts for phase change and natural convection. The optimization results yield a large acceleration of charge and discharge dynamics through three-dimensional geometries, specific convective features and optimized assemblies of periodic cellular materials. The second problem regards energy distribution with district heating networks. A fully deterministic robust design model and an adjoint-based control model are proposed, both coupled to a thermal and fluid-dynamic analysis framework constructed using a graph representation of the network. The numerical results demonstrate an increased resilience of the infrastructure thanks to particular connectivity layouts and its rapidity in handling mechanical failures. Finally, energy conversion with proton exchange membrane fuel cells is considered. An analysis model is developed that considers fluid flow, chemical species transport and electrochemistry and accounts for geometry modifications through a density-based description. The optimization results consist of intricate flow field layouts that promote both the efficiency and durability of the cell.
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Guidi, Mònica. „Micro RNA-Mediated regulation of the full-length and truncated isoforms of human neurotrophic tyrosine kinase receptor type 3 (NTRK 3)“. Doctoral thesis, Universitat Pompeu Fabra, 2009. http://hdl.handle.net/10803/7114.
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Neurotrophins and their receptors are key molecules in the development of thenervous system. Neurotrophin-3 binds preferentially to its high-affinity receptor
NTRK3, which exists in two major isoforms in humans, the full-length kinaseactive
form (150 kDa) and a truncated non-catalytic form (50 kDa). The two
variants show different 3'UTR regions, indicating that they might be differentially
regulated at the post-transcriptional level. In this work we explore how
microRNAs take part in the regulation of full-length and truncated NTRK3,
demonstrating that the two isoforms are targeted by different sets of microRNAs.
We analyze the physiological consequences of the overexpression of some of the
regulating microRNAs in human neuroblastoma cells. Finally, we provide
preliminary evidence for a possible involvement of miR-124 - a microRNA with no
putative target site in either NTRK3 isoform - in the control of the alternative
spicing of NTRK3 through the downregulation of the splicing repressor PTBP1.
Las neurotrofinas y sus receptores constituyen una familia de factores cruciales
para el desarrollo del sistema nervioso. La neurotrofina 3 ejerce su función
principalmente a través de una unión de gran afinidad al receptor NTRK3, del cual
se conocen dos isoformas principales, una larga de 150KDa con actividad de tipo
tirosina kinasa y una truncada de 50KDa sin dicha actividad. Estas dos isoformas
no comparten la misma región 3'UTR, lo que sugiere la existencia de una
regulación postranscripcional diferente. En el presente trabajo se ha explorado
como los microRNAs intervienen en la regulación de NTRK3, demostrando que las
dos isoformas son reguladas por diferentes miRNAs. Se han analizado las
consecuencias fisiológicas de la sobrexpresión de dichos microRNAs utilizando
células de neuroblastoma. Finalmente, se ha estudiado la posible implicación del
microRNA miR-124 en el control del splicing alternativo de NTRK3 a través de la
regulación de represor de splicing PTBP1.
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Lu, Chun-Han, und 呂俊翰. „Real-time Fluorescence Detection System for Cell Analysis“. Thesis, 2015. http://ndltd.ncl.edu.tw/handle/58921215391122022950.
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碩士國立屏東科技大學
車輛工程系所
103
In this study, a real-time fluorescence detection system for cell growth analysis is proposed utilizing the CMOS (Complementary Metal-Oxide-Semiconductor) modules with blue LEDs. The RAW 264.7 murine macrophage cell labels with CFSE (Carboxyfluorescein succinimidyl ester) fluorescence dye and the blue LEDs are for cells fluorescence excitation for real time cell growth fluorescence monitoring. The CMOS modules and LEDs are controlled using custom-made LABVIEW code to capture real time cell growth fluorescence images automatically. The cell growth images are analysis by the NI Vision program. In the proposed system, the cell proliferation was evaluated by doubling time and cells division index in the cells growth processes. In this study, the doubling time and cells division index of RAW264.7 cell are 24 hr, 66.59%. Compare to the traditional hemocytometer cells counter, the doubling time and cells division index of RAW264.7 cell are 17.2 hr, 68.75%. The fluorescence intensities decay in the doubling time are 33.8%, 41.3%, 46.2% and 50%, respectively. The CMOS module magnification rate is about 40 x to 50 x compare to the traditional microscope. As a result, the proposed system provides an ideal solution for cell proliferation real time fluorescence monitoring applications in the bio-medicine field.
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Hsu, Che Wei, und 許哲瑋. „Development of a microfluidic chip for perfusion three-dimensional cell culture and real-time dynamic cell growth analysis“. Thesis, 2013. http://ndltd.ncl.edu.tw/handle/23123445306536155343.
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碩士長庚大學
醫療機電工程研究所
101
A perfusion three dimensional (3D) cell culture microfluidic chip has been developed for the real-time and non-invasive impedimetric monitoring of cell dynamics. The chip can continuously estimate the cell proliferation or chemosensitivity under different culture conditions. It is capable of onsite detecting the cell number or viability in the 3D cell culture construct without sacrificing the cultured cells. The chip consists of a substrate, a cover layer and a fluidic layer. The chip substrate is a glass substrate and a pair of 3D vertical electrodes is fabricated on its surface. The cover layer and fluidic layer are made of poly¬dimethylsiloxane (PDMS) material and formed by soft lithography. A culture chamber is defined by the openings of the cover layer and the fluidic layer provides fluidic connection for medium perfusion purpose. By bonding these layers, the microfluidic chip can be fabricated. For the fabrication of the 3D vertical electrodes, planar electrodes were first fabricated on the substrate by standard micro-fabrication techniques. Then, the cover layer was bonded to the substrate with the alignment of the culture chamber and the planar electrodes. By copper electroplating process, a pair of 3D vertical electrodes were grown from the planar electrodes and located at the opposite sidewalls of the culture chamber. In this study, human oral cancer cell-line (OEC-M1) were used and encapsulated in 3D agarose scaffold and cultured in the microfluidic chip. On-site impedance measurement was performed to estimate the cellular response, i.e., cell number and viability, in the 3D culture construct. Cell number in the 3D construct was shown to be proportional to the impedance magnitude of the entire construct. Therefore, perfusion 3D cell culture was performed for up to 5 days and cell proliferation can be monitored by the impedimetric analysis. Furthermore, real-time monitoring of cell viability under the perfusion of anti-cancer drug in different concentrations was conducted and the impedance magnitude was directly correlated with the cell viability. From the confirmation of the endpoint cell viability assays, a concentration-dependent effect was shown; however, the response of cell viability during the drug treatment was able to be traced by the impedance measurement. This work showed cell proliferation and chemosensitivity in 3D cell culture format can be monitored by impedance measurement. This microfluidic chip has a high potential to develop a useful analytical platform for cancer research.
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Chang, Chia-Hao, und 張家豪. „Evaluation of digital real-time PCR assay as a molecular diagnostic tool for single-cell analysis“. Thesis, 2018. http://ndltd.ncl.edu.tw/handle/ycm2d3.
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博士國立臺灣大學
高分子科學與工程學研究所
106
In a single-cell study, isolating and identifying single cells are essential, but these processes often require a large investment of time or money. The aim of this study was to isolate and analyse single cells using a novel platform, the PanelChip™ Analysis System, which includes 2500 microwells chip and a digital real-time polymerase chain reaction (dqPCR) assay, in comparison with a standard RT-PCR (qPCR) assay. Through the serial dilution of a known concentration standard, namely pUC19, the accuracy and sensitivity levels of two methodologies were compared. The two systems were also tested on the basis of expression levels of the genetic markers vimentin, E-cadherin, N-cadherin and GAPDH in A549 lung carcinoma cells at two known concentrations. Furthermore, the influence of a known PCR inhibitor commonly found in blood samples, heparin, was evaluated in both methodologies. Finally, mathematical models were proposed and separation method of single cells was verified; moreover, gene expression levels during epithelial–mesenchymal transition in single cells under TGFβ1 treatment were measured. The drawn conclusion is that dqPCR performed using PanelChip™ has good linearity (R^2=0.9974) at 3.4 to 3.4 x 10^8 copies/μL, and it is superior to the standard qPCR in terms of, reproducibility (at 2pg/μL, dqPCR CV:1.97 < qPCR CV:3.94 ; at 200pg/μL , dqPCR CV:1.30 < qPCR CV:8.11) and heparin tolerance (dqPCR IC50: 0.02IU/mL > qPCR IC50: 0.002IU/mL). The dqPCR assay is a potential tool for clinical diagnosis and single-cell applications.
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Malik, Neelam. „Gene expression analysis of squamous cell carcinoma of the oesophagus using a novel real time PCR probe system“. Thesis, 2010. http://hdl.handle.net/10413/706.
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Squamous cell carcinoma of the oesophagus (OSCC) is a common malignancy that occurs with high frequency in certain parts of the world, including South Africa. The aetiology of OSCC has remained unclear although many studies suggest that it is caused by a combination of variable risk factors. Recent reports implicate a variety of genetic factors in the carcinogenesis of OSCC but their involvement is yet to be defined.Thesis (M.Med)-University of KwaZulu-Natal, Durban, 2010.
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Yuan, Fan. „Optimization of microelectrode sensor sensitivity for real-time monitoring important physiological parameters of human renal epithelial cell“. Thesis, 2020. https://hdl.handle.net/2144/40715.
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In order to calculate specific impedance of cell-covered electrodes in a Equation of morphological parameters of cell per se, an ECIS model of Human Renal Epithelial Cell are created by analysis partial differential equations describing three intrinsic pathways of electrical currents in the system. Based on this cell model, this research explores how some adjustable dimensional parameters of electrode-configuration impact sensor sensitivity by changing the overall impedance contribution of electrical double layer. Namely, it includes electrode planner area, spacing between working and counter electrode and geometry of electrode, scanning frequency. Qualitative studies on how sensor sensitivity rely on configurational parameters are conducted with these parameters involved. Moreover, theoretical analysis of sensitivity by using equivalent circuit model is also carried out. As results of COMSOL simulations, special double layer electrode configurations and selectively planted cell monolayer arrangement are proposed regardless of fabrication difficulties. Accordingly, some possible strategies to make these arrangements come true are also illustrated. Finally, superior possible COMSOL simulation model is suggested and discussed for future optimization works.2021-05-07T00:00:00Z
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LIN, YU-JEN, und 林俞任. „Investigation on Equivalent Circuit Analysis of Large-area Arrayed TiO2/ZnO Dye-sensitized Solar Cell Modified by Graphene Integrated with Magnetic Beads and Study on Wireless-based Remote Real-time Monitoring System“. Thesis, 2016. http://ndltd.ncl.edu.tw/handle/rsa2ju.
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Kuo, Chien-Hung, und 郭建宏. „Investigation on Photovoltaic Properties of Flexible Arrayed Dye-sensitized Solar Cell Based on IGZO/ TiO2 Double Layered Structure Modified by Graphene under the Low Illumination and Study on Impedance Analysis and Wireless-based Remote Real-time Monitoring System“. Thesis, 2018. http://ndltd.ncl.edu.tw/handle/a6xv27.
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碩士國立雲林科技大學
電子工程系
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In this thesis, a way to improve the photovoltaic conversion efficiency (η) of dye-sensitized solar cell (DSSC) has been provided. The structure was divided into two parts. In the first part, the reduced graphene oxide (RGO) - TiO2 composite was fabricated by using hydro-thermal method, which was acted as the dye - adsorbed layer. In the second part, the indium gallium zinc oxide (IGZO) was deposited between dye-adsorbed layer and electrolyte by using sputter system. The DSSC was investigated by electrochemical impedance spectroscopy (EIS), sun light simulation system, field emission scanning electron microscopy (SEM), energy dispersive spectrometer (EDS), ultraviolet-visible spectrophotometer (UV-Visible), X-ray photoelectron spectroscopy (XPS)/electron spectroscopy for chemical analysis (ESCA), X-ray diffractometer (XRD), Raman spectroscopy and transmission electron microscope (TEM). We investigated the photovoltaic properties, series-parallel connection module, internal interface impedance, surface morphology and energy band diagram of arrayed dye-sensitized solar cell based on RGO - TiO2 /IGZO photoelectrode under low illumination. According to the experimental results, due to the high mobility of RGO, which acted as a bridge and accelerated the electron transportation from conduction band of titanium dioxide to conduction band of fluorine doped tin oxide (FTO) glass. That was to say, probability of electron recombination between photo-generated electrons and oxidized-dye molecule was reduced. Furthermore, the energy band gap of dye-adsorbed layer decreased after introducing RGO, which could extend the wavelength range of absorbed-light. Particularly, the amount of harvesting-light is increased. In addition, the high specific surface of RGO was able to increase the amount of dye-loading. The IGZO film was acted as an energy barrier to prevent I-3 from recombining with electrons, which means that it could reduce the probability of reverse recombination. Those modifications of photoelectrode could improve the short-circuit current density (Jsc) of DSSC. Because the photo-generated electrons were reduced with decrease in illumination intensity, that indicated the scattering among electrons was reduced. In order words, the photoluminescence quantum yield (PLQY) will be increased, and the photovoltaic conversion efficiency of DSSC could increase under lower illumination intensity. Finally, the device was investigated by using the wireless-based remote real-time monitor, stability and life-time by source measure unit (SMU) and LabVIEW from National Instruments.
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Shimoga, Muddappa Vinay Kumar. „Electrochemical model based condition monitoring of a Li-ion battery using fuzzy logic“. Thesis, 2014. http://hdl.handle.net/1805/5588.
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Indiana University-Purdue University Indianapolis (IUPUI)There is a strong urge for advanced diagnosis method, especially in high power battery packs and high energy density cell design applications, such as electric vehicle (EV) and hybrid electric vehicle segment, due to safety concerns. Accurate and robust diagnosis methods are required in order to optimize battery charge utilization and improve EV range. Battery faults cause significant model parameter variation affecting battery internal states and output. This work is focused on developing diagnosis method to reliably detect various faults inside lithium-ion cell using electrochemical model based observer and fuzzy logic algorithm, which is implementable in real-time. The internal states and outputs from battery plant model were compared against those from the electrochemical model based observer to generate the residuals. These residuals and states were further used in a fuzzy logic based residual evaluation algorithm in order to detect the battery faults. Simulation results show that the proposed methodology is able to detect various fault types including overcharge, over-discharge and aged battery quickly and reliably, thus providing an effective and accurate way of diagnosing li-ion battery faults.