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Cereto, Massagué Adrià. "Development of tools for in silico drug discovery". Doctoral thesis, Universitat Rovira i Virgili, 2017. http://hdl.handle.net/10803/454678.
Pełny tekst źródłaEl cribado virtual es un método quimioinformático que consiste en la criba de moléculas bioactivas de grandes bases de datos de moléculas pequeñas. Esto permite a los investigadores ahorrarse el coste de probar experimentalmente cientos o miles de compuestos candidatos, reduciéndolos hasta cantidades manejables. Para la validación de los métodos de cribado virtual hacen falta bibliotecas de moléculas señuelo. El software DecoyFinder fue desarrollado como aplicación gráfica de fácil uso para la construcción de bibliotecas de moléculas señuelo, y fue posteriormente ampliado con los hallazgos de investigación posterior sobre la construcción i rendimiento de bibliotecas de moléculas señuelo. El Protein Data Bank (PDB) es muy útil porque proporciona estructuras tridimensionales para complejos proteina-ligando, y por tanto, información sobre como interactúan. Para los métodos de cribado virtual que dependen de ellas, es extremadamente importante su fiabilidad. VHELIBS fue desarrollado como herramienta para inspeccionar e identificar, fácil e intuitivamente, las estructuras fiables del PDB, basándose en como de bueno es su encaje con sus correspondientes mapas de densidad electrónica. Mientras que el cribado virtual intenta encontrar nuevas moléculas bioactivas para determinadas dianas, el enfoque inverso también se utiliza: a partir de una molécula, buscar dianas donde presente actividad biológica no documentada. Este cribado inverso es conocido en inglés como “in silico target fishing”, o pesca de dianas “in silico”, y es especialmente útil en el ámbito de la reutilización de fármacos. Al comenzar esta tesis, no había ninguna plataforma de “target fishing” de libre acceso, y aunque durante los años se han desarrollado algunas, en todos los casos su predicción de bioactividad es cualitativa. Por eso se desarrolló una plataforma propia de “target fishing” de libre acceso, con la implementación de un nuevo método que proporciona la primera predicción cuantitativa de bioactividad para este tipo de plataforma.
Virtual screening is a cheminformatics method that consists of screening large small-molecule databases for bioactive molecules. This enables the researcher to avoid the cost of experimentally testing hundreds or thousands of compounds by reducing the number of candidate molecules to be tested to manageable numbers. For their validation, virtual screening approaches need decoy molecule libraries. DecoyFinder was developed as an easy to use graphical application for decoy library building, and later updated after some research into decoy library building and their performance when used for 2D similarity approaches. The Protein Data Bank (PDB) is very useful because it provides 3D structures for protein-ligand complexes and, therefore, information on how certain ligands bind and interact with their targets. For virtual screening apporaches relying on these structures, it is of the utmost importance that the data available on the PDB for the ligand and its binding site are reliable. VHELIBS was developed as a tool to easily and intuitively inspect and identify reliable PDB structures based on the goodness of fitting between ligands and binding sites and their corresponding electron density map. While virtual screening aims to find new bioactive molecules for certain targets, the opposite approach is also used: starting from a given molecule, to search for a biological target for which it presents previously undocumented bioactivity. This reverse screening is known as in silico or computational target fishing or reverse pharmacognosy, and it is specially useful for drug repurposing or repositioning. When this thesis was started, there were no freely available target fishing platforms, but some have been developed during the years. However, they are qualitative in the nature of their activity prediction, and thus we set out to develop a freely accessible target fishing web service implementing a novel method which provides the first quantitative activity prediction: Anglerfish.
Islam, R. S. "Novel engineering tools to aid drug discovery processes". Thesis, University College London (University of London), 2007. http://discovery.ucl.ac.uk/1444794/.
Pełny tekst źródłaHesping, Eva M. "New inhibitors and tools to advance HDAC drug discovery for malaria". Thesis, Griffith University, 2021. http://hdl.handle.net/10072/403646.
Pełny tekst źródłaThesis (PhD Doctorate)
Doctor of Philosophy (PhD)
School of Environment and Sc
Science, Environment, Engineering and Technology
Full Text
Jenkins, Michael Joseph. "Decisional tools for cost-effective bioprocess design for cell therapies and patient-specific drug discovery tools". Thesis, University College London (University of London), 2018. http://discovery.ucl.ac.uk/10046409/.
Pełny tekst źródłaCarrascosa, Baena María Carmen 1972. "Next generation of informatics tools for big data analytics in drug discovery". Doctoral thesis, Universitat Pompeu Fabra, 2018. http://hdl.handle.net/10803/586011.
Pełny tekst źródłaThe classical silver bullet paradigm of one drug interacting with a single target linked to a disease is currently challenged. It is now widely recognized that one drug interacts with multiple targets and these targets are involved in many biological pathways and expressed in a variety of organs. As the notion of complexity has been gradually accepted, the reductionist drug discovery approach has naturally evolved towards systems multilevel strategies. Thanks to technological advances, there has been a huge increase of data generated in the various fields relevant to drug discovery, namely, chemistry, pharmacology, toxicology, genomics, metabolomics, etc., which has expanded dramatically our ability to generate computational models with increasing performance and coverage. But ultimately, extracting knowledge from this complex, vast and heterogeneous amount of data is not straightforward. The main objective of this Thesis is to develop new interactive analytics and visualization tools and investigate their ability to extract knowledge from highly interconnected data when implemented into an integrated flexible platform to facilitate drawing simple answers from complex questions. In particular, special emphasis will be put in the navigation aspects of the relationships between systemic entities (small molecules and their metabolite, protein targets, safety terms).
Cornet, Bartolomé Carles 1991. "Novel tools in drug discovery : optimising the use of zebrafish for assessing drug safety and antitumoral efficacy". Doctoral thesis, Universitat Pompeu Fabra, 2020. http://hdl.handle.net/10803/668470.
Pełny tekst źródłaLa alta tasa de deserción de medicamentos durante fases clínicas y posteriores a la comercialización es uno de los factores principales que contribuyen a la crisis de productividad que afecta a la industria farmacéutica hoy en día. Este problema es especialmente preocupante en el sector del cáncer, donde es de dos a cuatro veces mayor que en otros sectores de la salud. La mayoría de estos medicamentos son descartados debido a problemas de seguridad (principalmente cardio, neuro, y hepatotoxicidad) y de eficacia, lo que reflejan las limitaciones de los modelos preclínicos actuales para anticipar tales inconvenientes. En este contexto, se necesitan nuevos modelos para abordar este problema y cumplir con las nuevas demandas (mayor rendimiento y predictividad) de los procesos de investigación y desarrollo (I+D). El pez cebra es un vertebrado con alta homología con los humanos y propiedades biológicas únicas, que lo hacen adecuado para estudios de alto rendimiento. El objetivo final de mi tesis es mejorar el uso de este modelo animal en un intento de mejorar la eficiencia general del proceso de I+D y, así, aliviar la crisis de productividad. Primero, se ha generado una metodología de rendimiento medio para la evaluación in vivo de las toxicidades cardíaca, neuronal, y hepática en un mismo animal, en línea con en el principio de las 3Rs. En segundo lugar, se ha estandardizado, validado y automatizado, el xenotrasplante de células tumorales humanas en larvas de pez cebra para el estudio de la eficacia de fármacos antitumorales. Los resultados obtenidos ayudan a consolidar y validar el uso del pez cebra en el proceso de I+D de nuevos fármacos, como puente entre los modelos in vitro y los modelos in vivo de mamíferos.
Mezzanotte, Laura <1982>. "Bioanalytical applications of multicolour bioluminescence imaging: new tools for drug discovery and development". Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2011. http://amsdottorato.unibo.it/3536/1/Mezzanotte_Laura_TESI.pdf.
Pełny tekst źródłaMezzanotte, Laura <1982>. "Bioanalytical applications of multicolour bioluminescence imaging: new tools for drug discovery and development". Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2011. http://amsdottorato.unibo.it/3536/.
Pełny tekst źródłaSantiago, Daniel Navarrete. "Use and Development of Computational Tools in Drug Discovery: From Small Molecules to Cyclic Peptides". Scholar Commons, 2012. http://scholarcommons.usf.edu/etd/4398.
Pełny tekst źródłaPatel, Hitesh [Verfasser], i Irmgard [Akademischer Betreuer] Merfort. "Use and development of chem-bioinformatics tools and methods for drug discovery and target identification". Freiburg : Universität, 2015. http://d-nb.info/1115495917/34.
Pełny tekst źródłaLangron, E. "Drug-discovery tools for cystic fibrosis : optical probes to quantify gating and trafficking of ΔF508-CFTR". Thesis, University College London (University of London), 2016. http://discovery.ucl.ac.uk/1538676/.
Pełny tekst źródłaCOLOMBO, ELEONORA. "RATIONAL DESIGN, SYNTHESIS AND NANOTECHNOLOGIES AS TOOLS IN EARLY DRUG DISCOVERY: CANCER AND NEURODEGENERATION AS TARGETS". Doctoral thesis, Università degli Studi di Milano, 2021. http://hdl.handle.net/2434/818993.
Pełny tekst źródłaAbshire, James R. (James Robbins). "Development of novel chemical biology tools to probe malaria parasite physiology and aid in antimalarial drug discovery". Thesis, Massachusetts Institute of Technology, 2015. http://hdl.handle.net/1721.1/98921.
Pełny tekst źródłaThis electronic version was submitted by the student author. The certified thesis is available in the Institute Archives and Special Collections.
Cataloged from student-submitted PDF version of thesis.
Includes bibliographical references.
Malaria remains a major burden to global public health. Antimalarial drugs are a mainstay in efforts to control and eventually eradicate this disease. However, increasing drug resistance threatens to reverse recent gains in malaria control, making the discovery of new antimalarials critical. Antimalarial discovery is especially challenging due to the unique biology of malaria parasites, the scarcity of tools for identifying new drug targets, and the poorly understood mechanisms of action of existing antimalarials. Therefore, this work describes the development of two chemical biology tools to address unmet needs in antimalarial drug discovery. A particular challenge in antimalarial development is a shortage of validated parasite drug targets. Potent antimalarials with demonstrated clinical efficacy, like the aminoquinolines and artemisinins, represent a promising basis for rational drug development. Unfortunately, the molecular targets of these drugs have not been identified. While both are thought to interact with parasite heme, linking in vitro heme binding with drug potency remains challenging because labile heme is difficult to quantify in live cells. This work presents a novel genetically-encoded heme biosensor and describes its application to quantify labile heme in live malaria parasites and test mechanisms of antimalarial action. Another challenge is posed by the widespread malaria parasite Plasmodium vivax, which, unlike P. falciparum, cannot be propagated in vitro, hindering research into parasite biology and drug target identification. P. vivax preferentially invades reticulocytes, which are impractical to obtain in continuous supply. The basis for this invasion tropism remains incompletely understood, mainly because current tools cannot directly link molecular binding events to invasion outcomes. This work presents novel methods for immobilizing synthetic receptors on the red blood cell surface. These receptors are used in proof-of-concept experiments to investigate requirements for efficient invasion via a well-characterized P. falciparum invasion pathway, suggesting this method can be used to elucidate molecular mechanisms underlying parasite invasion tropisms. Future receptor designs could promote the invasion of P. vivax into mature red blood cells and potentially facilitate practical in vitro culture. Taken together, these tools present new opportunities for drug discovery to aid efforts in malaria control and eventual eradication.
by James R. Abshire.
Ph. D.
Gee, Sally. "The case of research tools for drug discovery : evolving user-producer inter-dependencies and the exchange of knowledge". Thesis, University of Manchester, 2007. http://www.manchester.ac.uk/escholar/uk-ac-man-scw:80887.
Pełny tekst źródłaKamal, Ahmed [Verfasser], i Rolf W. [Akademischer Betreuer] Hartmann. "From Small Organic Molecules to Peptides As Tools For Anti-infectives Drug Discovery / Ahmed Kamal ; Betreuer: Rolf W. Hartmann". Saarbrücken : Saarländische Universitäts- und Landesbibliothek, 2017. http://d-nb.info/1152095374/34.
Pełny tekst źródłaKamal, Ahmed Verfasser], i Rolf W. [Akademischer Betreuer] [Hartmann. "From Small Organic Molecules to Peptides As Tools For Anti-infectives Drug Discovery / Ahmed Kamal ; Betreuer: Rolf W. Hartmann". Saarbrücken : Saarländische Universitäts- und Landesbibliothek, 2017. http://d-nb.info/1152095374/34.
Pełny tekst źródłaHsing, Michael. "Developing bioinformatics tools and analyses on protein indels and protein-protein interactions : novel applications for drug discovery in Staphylococcus aureus". Thesis, University of British Columbia, 2010. http://hdl.handle.net/2429/18714.
Pełny tekst źródłaKhashan, Raed Saeed Tropsha Alexander. "Development and application of ligand-based and structure-based computational drug discovery tools based on frequent subgraph mining of chemical structures". Chapel Hill, N.C. : University of North Carolina at Chapel Hill, 2007. http://dc.lib.unc.edu/u?/etd,1243.
Pełny tekst źródłaTitle from electronic title page (viewed Mar. 26, 2008). " ... in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the School of Pharmacy (Division of Medicinal Chemistry and Natural Products)." Discipline: Medicinal Chemistry and Natural Products; Department/School: Pharmacy.
Yglesias, Tatiana. "Application of process analysis and optimization tools in hit-to-lead and lead optimization phases of drug discovery at EPP, NIBR". Thesis, Massachusetts Institute of Technology, 2010. http://hdl.handle.net/1721.1/59882.
Pełny tekst źródłaCataloged from PDF version of thesis.
Includes bibliographical references (p. 82-84).
Given that research is based on innovation, it has been believed that its activities can only be optimized with equipment upgrade, increment in personnel scientific knowledge, development of new analytical software and/or changing the areas of study. After realizing the limited results achieved with these approaches, lab representatives started to notice the opportunity of introducing process optimization tools, such as Lean and Six Sigma, which showed success in manufacturing environments,. This project analyzes the interrelation between process and results, providing a clear explanation of cause and effect conditions, and a concise list of areas for improvement. Specifically, the document defines a measurement system using process maps and key performance indicators (KPIs). With this, the document describes the current state through historic trends, provides a complete data and root cause analysis for current state description, and provides a process capability study for the available indicators. Implementation of the steps mentioned above show how focus in lab turnaround times have been deviating attention from more impactful improvements, which can greatly affect overall drug discovery duration. Also, the analysis identifies that constant technology changes caused constant adaptation of process procedures, which generated non-value added activities. These non-value added activities today occupy about 50% of a lab associate's time. Lastly, historic data evaluation shows that root cause statistical analysis is limited by the presence of a combination of special and common cause variations. Some of the project recommendations include: incorporation of chemist's knowledge about compound potency, integration of equipment and software information, change in booking system, incorporation of assay and plate criteria, definition of standard procedures for specific activities, and integration of assay development and data submission tools. Overall, these changes can lead to a 50% reduction in the profiling times greater than 60 days, decrease of 62% and 60% in Compound Manager (CM) and Compound Profiler (CP) non-value added times respectively, 30% decrease in CM and CP total duration per assay plate, and increase in profiling time stability and predictability. Despite the fact that timing and scale of available resources will impact the realized benefits, the proposed framework gives EPP the opportunity to assess the improvements by their effect and alignment with goals.
by Tatiana Yglesias.
M.B.A.
S.M.
Goldflam, Michael. "Combined use of NMR and computational tools for fragment based drug discovery targeting protein-protein interactions VEGF protein surface recognition as a case study". Doctoral thesis, Universitat de Barcelona, 2013. http://hdl.handle.net/10803/123711.
Pełny tekst źródłaEn el contexto de la presente tesis hemos abordado los siguientes objetivos: 1. El uso de métodos de RMN, basados tanto en la observación del ligando como en la observación de proteína, para estudiar la unión de los compuestos de una quimioteca a la zona de VEGF involucrada en la unión a sus receptores. La interacción VEGF/VEGFR puede ser considerada como un caso de estudio para la evaluación de las interfaces proteína-proteína mediante cribado de fragmentos. 2. Desarrollar herramientas basadas en la combinación de RMN y métodos computacionales para abordar: i) un sistema automático de diseño de mezclas de fragmentos; ii) el análisis automático de datos procedentes de cribados basados en RMN; iii) la evolución de fragmentos con muy baja afinidad. 3. Explorar el uso de técnicas de “mRNA display” para el descubrimiento de nuevos ligandos peptídicos para VEGF.
Guidotti, Laura <1987>. "Design and synthesis of small molecules as tools for drug discovery and chemical biology: identification of new LDH-A inhibitors and development of fluorescent probes to investigate neurogenesis". Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2018. http://amsdottorato.unibo.it/8438/1/Laura%20Guidotti%20Tesi.pdf.
Pełny tekst źródłaHatherley, Rowan. "Structural bioinformatics studies and tool development related to drug discovery". Thesis, Rhodes University, 2016. http://hdl.handle.net/10962/d1020021.
Pełny tekst źródłaHendry, Adam. "Xenopus laevis as a chemical genetic screening tool for drug discovery and development". Thesis, University of East Anglia, 2014. https://ueaeprints.uea.ac.uk/49595/.
Pełny tekst źródłaAnderson, Gregory William. "Critically derived human neural stem cell line as a potential tool for neuropsychiatric drug discovery". Thesis, King's College London (University of London), 2012. https://kclpure.kcl.ac.uk/portal/en/theses/critically-derived-human-neural-stem-cell-line-as-a-potential-tool-for-neuropsychiatric-drug-discovery(aa19f8bb-217d-46f7-9da4-782fcbed0ab9).html.
Pełny tekst źródłaMeiby, Elinor. "Progress of Weak Affinity Chromatography as a Tool in Drug Development". Doctoral thesis, Linnéuniversitetet, Institutionen för kemi och biomedicin (KOB), 2013. http://urn.kb.se/resolve?urn=urn:nbn:se:lnu:diva-25970.
Pełny tekst źródłaEtt läkemedel utövar sin funktion genom att påverka aktiviteten hos ett protein i kroppen då det binder till dess aktiva säte. Förändringen i aktivitet leder till fysiologiska förändringar i kroppen beroende på vilken funktion proteinet har. Med läkemedelsmolekyl avses här en liten organisk molekyl. Fragment-baserad läkemedelsutveckling är en ny metod for att ta fram nya läkemedel. Metoden fungerar genom att man bygger läkemedelsmolekyler utifrån mindre fragment som binder till målproteinet. Fragmenten hittar man genom att screena hela bibliotek av olika fragment mot samma målprotein för att urskilja de som binder till proteinets aktiva säte. Fördelen med den här metoden är bl. a. att med mindre molekyler som utgångspunkt kan en större del av antalet möjliga kombinationer av atomer representeras med ett mindre antal fragment än för större molekyler. Normalt utgörs ett fragmentbibliotek enbart av några hundra till några tusen substanser. Eftersom fragmenten är små har de få interaktionspunker och binder relativt svagt. De svaga bindningarna är svåra att se och mycket känsliga metoder behövs. Svagaffinitetskromatografi är en vätskekromatografisk metod som utvecklades för att studera svaga men mycket selektiva bindningar mellan biomolekyler. Den här avhandlingen syftar till att utveckla metoden för olika användningsområden inom läkemedelsutveckling, främst som en ny metod för fragment-screening. Här mäter man interaktionen mellan ett protein och ett fragment. Proteinet kopplas till ett material som sedan packas i en kolonn i formen av en cylinder. När provet pumpas igenom kolonnen kommer de analyter med affinitet till proteinets aktiva säte att fördröjas på kolonnen i relation till hur starkt de interagerar med målproteinet. I den här avhandlingen presenteras fragment-screening med svagaffinitetskromatografi gentemot ett antal olika typer av målproteiner. Resultatet överensstämmer väl med andra metoder för fragment-screening. Analys av reaktionsblandningar med svagaffinitetskromatografi demonstreras också. Därmed kan bindningen mellan en produkt i en reaktionsblandning och ett målprotein mätas direkt utan föregående uppreningssteg av reaktionsblandningen. Lipodiskar är små diskformade modellmembran som kan användas för att bl. a. mäta hur effektivt läkemedlet tas upp i kroppen vid behandling. Ett system med immobiliserade lipodiskar i en kolonn utvecklades med det framtida målet att kunna arbeta med membranproteiner med svagaffinitetskromatografi. Detta arbete utgör en del i att utveckla svagaffinitetskromatografi som en lättillgänglig och relativt billig metod för användning inom industrin och akademin för läkemedelsutveckling.
Christopeit, Tony. "Protein Interaction Studies with Low Molecular Weight Ligands : Applications for Drug Discovery, Basic Research and Diagnostic Tool Design". Doctoral thesis, Uppsala universitet, Institutionen för kemi - BMC, 2013. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-188328.
Pełny tekst źródłaNaidoo, Jerolen. "Functional miRNA-based Phenotypic Screening as a tool to delineate HIV-host interactions and facilitate Novel Drug Discovery". Thesis, Faculty of Health Sciences, 2021. http://hdl.handle.net/11427/33227.
Pełny tekst źródłaSchoepe, Stefanie. "Investigations of in vitro test systems for the detection of Glucocorticoid-induced skin atrophy as a tool in drug discovery". Doctoral thesis, Humboldt-Universität zu Berlin, Mathematisch-Naturwissenschaftliche Fakultät I, 2009. http://dx.doi.org/10.18452/15967.
Pełny tekst źródłaTopical glucocorticoids (GCs) are effective for the therapy of inflammatory skin diseases. However, their use is limited by their side effect potential, with skin atrophy being the most prominent one. Thus, determining the atrophogenic potential of novel compounds is of importance for drug development. Currently, there are no according predictive in vitro models available. The aim of this study was to establish such atrophy models. Rodent and human cutaneous cell types (3T3 cells, rat fibroblasts, HaCaT cells, human keratinocytes [NHEK] and fibroblasts) and human full-thickness skin equivalents (CellSystems AST-2000 and Phenions FTSM) were investigated. Atrophy markers related to proliferation, collagen metabolism and epidermal thickness were measured on mRNA, protein and cellular level, respectively. Additionally, by gene expression profiling of GC-treated rodent skin novel potential markers were identified, but subsequently not confirmed in vitro. After pilot studies 3 models were selected and treated with reference GCs for evaluation experiments: 1.) MMP1, -2, -3 and -9 mRNA expression in NHEK, 2.) COL1A1 and COL3A1 mRNA expression in 3T3 cells, 3.) epidermal thickness, collagen and MMP synthesis in FTSM. The read out parameters of all 3 test systems turned out to be regulated dose-dependently and correlated with the atrophogenic potential of the GCs. Finally, the predictability of the 3 recommended in vitro test system for the rodent in vivo situation was analyzed. In all 3 in vitro test systems, the treatment with a novel selective GC receptor agonist induced less atrophogenic effects than the reference GC clobetasol. Indeed, similar results were found in the hr/hr rat skin atrophy model. In summary, a cascade of 3 in vitro models is recommended to be applied for the characterization of the atrophogenicity of GC receptor ligands. Further experiments are necessary to eventually demonstrate the true predictability of these models for the clinical situation.
Dinh, Hong-Khanh Bao. "Gene expression profiling of thermal induced injury : a drug discovery and development tool for evaluating potential cytoprotective agents for the amelioration of thermal and laser injury to the eye /". Full text (PDF) from UMI/Dissertation Abstracts International, 2001. http://wwwlib.umi.com/cr/utexas/fullcit?p3008316.
Pełny tekst źródłaCheemakurthi, Usha Deepika. "DESIGN, DEVELOPMENT AND IMPLEMENTATION OF TOOLS IN". Thesis, 2010. http://hdl.handle.net/1805/2268.
Pełny tekst źródłaDr. Kelsey Forsythe,PhD, Chair; Dr. Malika Mahoui, PhD;Dr. David Wild, PhD
Spare, Lawson K. "The development of flow-assisted synthetic methodologies as tools for drug discovery". Thesis, 2020. http://hdl.handle.net/1959.7/uws:67212.
Pełny tekst źródłaAGARWAL, ANURAG. "COMPARATIVE ANALYSIS, GAP ANALYSIS AND OPTIMIZATION OF DRUG DISCOVERY TOOLS: A SYSTEMATIC EVALUATION FOR ENHANCED EFFICIENCY". Thesis, 2023. http://dspace.dtu.ac.in:8080/jspui/handle/repository/19988.
Pełny tekst źródłaFonseca, Maria Marques da Silva Mega da. "New in vitro models and synthetic biology tools for research and drug discovery against hepatotropic pathogens". Master's thesis, 2019. http://hdl.handle.net/10451/41699.
Pełny tekst źródłaThe lack of competent in vitro hepatic cell models has hindered the study of hepatotropic pathogens such as Hepatitis C virus (HCV) and consequently the development of new therapies. The main limitation is the shortage of a primary-like phenotype and functionality as well as limited permissiveness to infection. This work aimed at developing highly competent hepatic cell lines through lentiviral vector mediated immortalization of primary human hepatocytes (PHH). To this end, we started by developing a strategy to sustain lentiviral vector transduction of PHH by improving lentiviral vector design and optimizing transduction conditions. A dualcolor VCN PCR method was also implemented to titrate the immortalizing vectors with increased throughput. The data obtained and the tools established allowed the construction of immortalization libraries capable of driving appropriate expression. The functionality of one of these libraries was confirmed by qPCR and notable changes in morphology of PHH after one week of transduction. More, transduced PHH generated a heterogeneous population and cell clones. A total of 39 colonies was obtained by limiting dilution isolation. Proliferative cells that survived to expansion have demonstrated distinct morphologies confirming the ability of this strategy to generate biologically diverse cells. This high diversity will enable to dissect the determinants leading to the generation of cells permissive to infection and the identification of key factors underlying the hepatic primarylike phenotype. This work contributed to the creation of a platform for PHH immortalization by establishing a high throughput lentiviral vector production and titration methods compatible with use in PHH and by optimizing transduction conditions. A functional immortalization library was also constructed which led to the expansion of a heterogeneous population and 2 proliferating clones. With further optimization this platform will contribute to the establishment of new highly permissive and competent cells to support HCV entry and infection progression that ultimately can assist the efficient drug or vaccine development against HCV.
D'Aniello, Filomena. "Characterization of new bioactive natural products from marine sources as pharmaceutical tools and lead compounds in drug discovery processes". Tesi di dottorato, 2014. http://www.fedoa.unina.it/9845/1/d%27aniello%20filomena.pdf.
Pełny tekst źródłaMachado, Rita Biltes Lopes Gonçalves. "Optimization of a molecular cell-based sensor to detect PPARγ ligands - a tool for drug discovery in cyanobacteria". Master's thesis, 2021. https://hdl.handle.net/10216/136866.
Pełny tekst źródłaSchoepe, Stefanie [Verfasser]. "Investigations of in vitro test systems for the detection of Glucocorticoid-induced skin atrophy as a tool in drug discovery / von Stefanie Schoepe". 2009. http://d-nb.info/996968598/34.
Pełny tekst źródła