This thesis describes possibilities of measuring permittivity of various materials, especially with coaxial probe method. It describes character of measuring method, hardware requirements and how to calculate values from the measured. Objective of this thesis is to design measuring station for measuring permittivity of materials with coaxial probe including calibration set. Code in Matlab is also programmed and whole station is tested with known samples to discuss measurement accuracy.

The object of this work is to apply the measuring probe in the process of the machining and explore the use of the measuring probes. For the practical of the measurement was use measuring probes, the program supplied with the measuring probes, machining centers and the resulting data were evaluated on a PC. Finally, work is to evaluate the measurements and describe the appropriateness of the application.

The Aim of this Diploma thesis is the measurement on turbine cascade. The first part is focused on measurement of the flow characteristics of the turbine cascade using a three–hole pressure probe. The second part includes the blade profile pressure measurement. Both measurements are performed both experimentally and by computer simulation in Ansys and the results are compared. Part of the work is also file containining the computer simulation, which was carried out in CFX program.

This work is dedicated to small distance contact-less measurement. Especially the surface detection of the commutator or collector of the electrical machines was described. The probes utilising the induction principle of the distance measurement have been developed. The part of the work described the history of the induction distances sensing in FEEC BUT and the using of the induction principle in the industry small distances sensing nowadays. The paper concretely determines the possibilities of the small distances measurement using mentioned probes. The sensitivity for different materials was investigated and the error of the measurement for different measuring object was investigated, too. The effects, that could affected the measurement error, were described in the work. The contribution of the each effect for the total measuring error was specified. Next part is including the description of the measuring workstation and of the hardware and software. At the end the verification of the measuring principle and measuring workstation is presented. The verification has been made on three different commutator types. The verification results are in the conclusion.

This thesis deals with designing the probe used for measuring network flows. It contains theoretical analysis of network measurment topic, description of algorithms and principles used for network flow measurement. Emphasis on the probe architecture lies on efficient indexing algorithm and flow record flexibility, such that user is able to define format of flow record.

The doctoral thesis presents the diagnostic neutral lithium beam on the COMPASS tokamak in Prague. The technical part describes the system for injection of accelerated lithium beam into the COMPASS tokamak and the respective detection systems for beam emission spectroscopy. The physical part describes the analysis of measured spectroscopic data, with particular emphasis on density fluctuations in the edge plasma. Detailed analysis of turbulent structures outside of the confined plasma region is presented, as well as investigations of the ELM instabilities during high confinement mode plasma operation. The thesis also presents the development and proof-of-concept measurements of a new and unique diagnostic method - atomic beam probe. The method uses detection of the ionized part of the beam for current density profile measurements in the plasma edge.

Master's thesis applies theme unknown landscape. The theoretical part is devoted to concept landscape, which is viewed from different perspectives in the work; from defining the landscape as a generic term through subjective perception to the possibility of its use as an imaginative space. The research part describes field research, which deals with issues of different interpretations of landscape perception by children of younger school age. Within the framework of the didactic part are presented the tasks in which the theme of an unknown country is reflected in various forms and are designed for art lessons in elementary schools. Finally practic part of my work comes out from theme unknown countryside and I try to express my own artistic expression. KEYWORDS Imagination, spatial concepts, space, unknown landscape, field research, children of younger school age, creative activity

This thesis deals with design and implementation of software architecture for Flexible FlowMon probe, accessories for monitoring high speed computer networks based on IP flows. The probe has been developed in project named Liberouter. There is described flow based monitoring and export formats NetFlow version 5, NetFlow version 9 and IPIFX, which are very widely used. The thesis contains description of hardware part of Flexible FlowMon probe including its requirements for software, which are the base of the whole software architecture. There is detailed description of that part of software architecture which was implemented during the work on this thesis.

Tesis por compendio / [EN] The selective and sensitive detection of single nucleotide variations is essential for early diagnosis, individualized therapy, and disease prognosis. The SARS-CoV-2 pandemic has highlighted the pressing need to develop reliable, rapid and simple detection methods for mass diagnosis. Likewise, there is a growing demand for genomic biosensors that are selective, multi-analyte and low-cost and allow the detection and identification of certain oncological biomarkers. This doctoral thesis has focused on the development of integrated systems of isothermal amplification, selective hybridization and optical biosensing for detection of point mutations in the PIK3CA, KRAS and BRAF genes associated with colorectal cancer. These predictive biomarkers are related to increased cell proliferation, apoptosis, and resistance to monoclonal antibody treatments (Cetuximab and Panitumumab). Isothermal DNA amplification methods have been explored, as they are particularly suitable for the development of a new generation of diagnostic devices aimed at supporting precision medicine. Specifically, isothermal amplification by recombinase-polymerase (RPA) has been selected as an alternative to detection methods that require unique infrastructures. In addition, its integration with bioanalytical platforms has been investigated, which represents a great advance for the simplification of biorecognition processes and their optical detection. This methodology has presented advantages over other DNA detection systems, in terms of portability and equipment, as well as reduction of test times and the possibility of performing diagnostic tests outside the laboratory. This doctoral thesis, framed in this context, is structured in 4 chapters: In chapter 1 a new variant of recombinase-polymerase amplification is presented, called RPA-blocked, based on the enrichment of minority alleles by introducing a blocking agent. In addition, this research has developed an analytical support consisting of a polycarbonate chip with covalently anchored allele-specific probes.The integration of the method has allowed the development of a portable system for the simultaneous genotyping of mutations in exons 9 and 20 of the PIK3CA gene. in cell lines and tumor tissues of cancer patients. In Chapter 2, the development of a genosensor that incorporates magnetic particles conjugated to allele-specific probes for the concentration and detection of the selective amplification product derived from RPA-blocked is described. With this genosensor, hybridization times and reaction volumes have been reduced. This approach has resulted in a portable and low-cost system for genotyping the KRAS gene applicable to solid tumor samples. Chapters 3 and 4 focus on the development of thermoplastic surfaces for the covalent anchoring of allele-specific probes mediated by dendrimeric molecules, with the aim of increasing the immobilization density. Thus, the covalent anchoring to activated polycarbonate and cycloolefin thermoplastic surfaces of allele-specific probes has been studied, mediated by carboxylic dendrimers through carbodiimide chemistry and by dendrons through the chemoselective reaction of the thiol-ino group. These multiplexed genosensors have allowed the genotyping of the V600 codon of the BRAF gene and the H1047 codon of the PIK3CA gene, in biopsied tissue samples. The research carried out in this thesis has given rise to new methodological contributions of interest based on obtaining integrated biosensor systems. These platforms will contribute to the development of massive diagnostic tools. / [ES] La detección selectiva y sensible de variaciones de nucleótido único es fundamental para el diagnóstico precoz, la terapia individualizada y el pronóstico de enfermedades. La pandemia SARS-CoV-2 ha puesto de manifiesto la necesidad acuciante de desarrollar métodos de detección fiables, rápidos y sencillos para el diagnóstico masivo. Asimismo, existe una demanda creciente de biosensores genómicos que sean selectivos, multianalito y de bajo coste y permitan detectar e identificar ciertos biomarcadores oncológicos.La presente tesis doctoral se ha centrado en el desarrollo de sistemas integrados de amplificación isoterma, hibridación selectiva y biosensado óptico para la detección de mutaciones puntuales en los genes PIK3CA, KRAS y BRAF asociadas al cáncer colorrectal. Estos biomarcadores predictivos se relacionan con un incremento de la proliferación celular, apoptosis y resistencia a tratamientos con anticuerpos monoclonales (Cetuximab y Panitumumab). En este contexto, se han explorado los métodos isotermos de amplificación de ADN, dado que son particularmente adecuados para el desarrollo de una nueva generación de dispositivos de diagnóstico dirigidos a apoyar la medicina de precisión. En concreto, se ha seleccionado la amplificación isoterma por recombinasa-polimerasa (RPA) como una alternativa a los métodos de detección que requieren de infraestructuras singulares. Además, se ha investigado su integración con plataformas bioanalíticas, lo que supone un gran avance para la simplificación de los procesos de biorreconocimiento y su detección óptica. Esta metodología ha presentado ventajas frente a otros sistemas de detección de ADN, en términos de portabilidad y equipos, así como reducción de tiempos de ensayo y la posibilidad de realizar las pruebas de diagnóstico fuera del laboratorio. La presente tesis doctoral, enmarcada en este contexto, se estructura en 4 capítulos: En el capítulo 1 se presenta una nueva variante de la amplificación por recombinasa-polimerasa, denominada RPA-bloqueada, basado en el enriquecimiento de alelos minoritarios mediante de la introducción de un agente bloqueante. Además, en esta investigación se ha desarrollado un soporte analítico formado por un chip de policarbonato con sondas alelo-específicas ancladas covalentemente.La integración del método ha permitido desarrollar un sistema portátil para el genotipado simultáneo de mutaciones en los exones 9 y 20 del gen PIK3CA en líneas celulares y en tejidos tumorales de pacientes oncológicos. En el capítulo 2, se describe el desarrollo de un genosensor que incorpora partículas magnéticas conjugadas a sondas alelo-específicas para la concentración y detección del producto de amplificación selectivo derivado de la RPA-bloqueada. Con este genosensor, se han reducido los tiempos de hibridación y los volúmenes de reacción. Esta aproximación se ha concretado en un sistema portátil y de bajo coste para el genotipado del gen KRAS aplicable a muestras de tumores sólidos. Los capítulos 3 y 4 se centran en el desarrollo de superficies termoplásticas para el anclaje covalente de sondas alelo-específicas mediado por moléculas dendriméricas, con el objetivo de incrementar la densidad de inmovilización. Así, se ha estudiado el anclaje covalente a superficies termoplásticas de policarbonato y cicloolefina activadas, de sondas alelo-específicas, mediado por dendrímeros carboxílicos mediante la química de la carbodiimida y por dendrones mediante la reacción quimioselectiva del grupo tiol-ino. Dichos genosensores multiplexados han permitido el genotipado del codón V600 del gen BRAF y el codón H1047 del gen PIK3CA, en muestras de tejido biopsiado. Las investigaciones desarrolladas en la presente tesis han dado lugar a nuevas aportaciones metodológicas de interés basadas en la obtención de sistemas biosensores integrados. Estas plataformas, contribuirán al desarrollo de herramientas de diag / [CAT] La detecció selectiva i sensible de variacions de nucleòtid únic és fonamental per al diagnòstic precoç, la teràpia individualitzada i el pronòstic de malalties. La pandèmia SARS-CoV-2 ha posat de manifest la necessitat apressant de desenvolupar mètodes de detecció fiables, ràpids i senzills per al diagnòstic massiu. Així mateix, existeix una demanda creixent de biosensors genòmics que siguen selectius, multianàlit i de baix cost i permeten detectar i identificar uns certs biomarcadors oncológics. La present tesi doctoral s'ha centrat en el desenvolupament de sistemes integrats d'amplificació isoterma, hibridació selectiva i biosensat òptic per a la detecció de mutacions puntuals en els gens PIK3CA, KRAS i BRAF associades al càncer colorectal. Aquests biomarcadors predictius es relacionen amb un increment de la proliferació cel·lular, apoptosi i resistència a tractaments amb anticossos monoclonals (Cetuximab i Panitumumab). S'han explorat els mètodes isoterms d'amplificació d'ADN, atés que són particularment adequats per al desenvolupament d'una nova generació de dispositius de diagnòstic dirigits a donar suport a la medicina de precisió. En concret, s'ha seleccionat l'amplificació isoterma per recombinasa-polimerasa (RPA) com una alternativa als mètodes de detecció que requereixen d'infraestructures singulars. A més, s'ha investigat la seua integració amb plataformes bioanalítiques, la qual cosa suposa un gran avanç per a la simplificació dels processos de biorreconeiximent i la seua detecció òptica. Aquesta metodologia ha presentat avantatges enfront d'altres sistemes de detecció d'ADN, en termes de portabilitat i equips, així com reducció de temps d'assaig i la possibilitat de realitzar les proves de diagnòstic fora del laboratori. La present tesi doctoral, emmarcada en aquest context, s'estructura en 4 capítols: En el capítol 1 es presenta una nova variant de l'amplificació per recombinasa-polimerasa, denominada RPA-bloquejada, basat en l'enriquiment d'al·lels minoritaris mitjançant de la introducció d'un agent bloquejant. A més, en aquesta investigació s'ha desenvolupat un suport analític format per un xip de policarbonat amb sondes al·lel-específiques ancorades covalentemente.la integració del mètode ha permés desenvolupar un sistema portàtil per al genotipat simultani de mutacions en els exons 9 i 20 del gen PIK3CA en línies cel·lulars i en teixits tumorals de pacients oncològics. En el capítol 2, es descriu el desenvolupament d'un genosensor que incorpora partícules magnètiques conjugades a sondes al·lel-específiques per a la concentració i detecció del producte d'amplificació selectiu derivat de la RPA-bloquejada. Amb aquest genosensor, s'han reduït els temps d'hibridació i els volums de reacció. Aquesta aproximació s'ha concretat en un sistema portàtil i de baix cost per al genotipat del gen KRAS aplicable a mostres de tumors sòlids. Els capítols 3 i 4 se centren en el desenvolupament de superfícies termoplàstiques per a l'ancoratge covalent de sondes al·lel-específiques mediat per molècules dendrimériques, amb l'objectiu d'incrementar la densitat d'immobilització. Així, s'ha estudiat l'ancoratge covalent a superfícies termoplàstiques de policarbonat i cicloolefina activades, de sondes al·lel-específiques, mediat per dendrimers carboxílics mitjançant la química de la carbodiimida i per dendrones mitjançant la reacció quimioselectiva del grup tiol-ino. Dits genosensors multiplexats han permés el genotipat del codó V600 del gen BRAF i el codó H1047 del gen PIK3CA, en mostres de teixit biopsiat Les investigacions desenvolupades en la present tesi han donat lloc a noves aportacions metodològiques d'interés basades en l'obtenció de sistemes biosensors integrats. Aquestes plataformes, contribuiran al desenvolupament d'eines de diagnòstic massiu. / Martorell Tejedor, S. (2021). Desarrollo de métodos integrados para la determinación de biomarcadores genéticos [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/176006 / Compendio

[pt] A Doença de Alzheimer (DA) é a maior causa de demência, com cerca de 50 milhões de pacientes em todo mundo em 2020, e este número é projetado para triplicar até 2050, com o custo estimado de 1 trilhão de dólares anualmente. Biometais como cobre e zinco em sua forma iônica são capazes de interagir com o peptídeo beta-amiloide (Abeta), relacionado à doença, deixando-o mais suscetível a agregação, além de contribuírem para o aumento do estresse oxidativo no organismo.A perturbação na homeostase metálica no cérebro, especialmente de cobre e zinco, aumenta o estresse oxidativo no cérebro, gerando espécies reativas de oxigênio (ERO) e nitrogênio (ERN) principalmente através de reações de Fenton e Haber-Weiss. Adicionalmente, cobre e zinco aceleram a agregação de A(beta) ao se ligarem em certos resíduos deste peptídeo.Neste contexto, ligantes conhecidos como metalóforos vêm sido desenvolvidos com objetivo de sequestrar os íons metálicos ligados ao peptídeo ou proteína na finalidade de reduzir os oligômeros de A(beta) e/ou estresse oxidativo induzido por metais. Invés de ligar e sistematicamente remover quaisquer metais dos tecidos, os metalóforos agem a fim de restaurar a homeostase metálica fisiológica através da ligação específica com cobre e/ou zinco, atenuando suas interações anormais com o peptídeo, assim prevenindo a oligomerização.Neste sentido, dois ligantes N-acil-hidrazônicos, com fração eletroativa contendo ferroceno, denominados Feizona e Ferfurona, foram sintetizados e caracterizados com técnicas como FTIR, (1)H NMR, TGA, UV-Vis e XRD. A complexação desses ligantes com Cu(2+) e Zn(2+), bem como sua desprotonação amídica, foram estudadas por voltametria cíclica. Estes ligantes apresentaram capacidade satisfatória em coordenar íons de Cu(2+) em solução em temperatura ambiente, porém necessitam de um meio fortemente básico para coordenarem espécies de Zn(2+). Esta característica indica uma seletividade para íons de Cu(2+)em meio fisiológico.Estudos de estabilidade hidrolítica foram avaliados por espectroscopia eletrônica, mostrando excelente estabilidade configurada pela conjugação eletrônica na estrutura dos compostos. Adicionalmente, a estabilidade fotolítica também foi avaliada por diferentes técnicas, mostrando-se dependente do solvente.O estudo de complexação por voltametria cíclica destes ligantes comprova a dependência de um meio básico para coordenação efetiva de zinco, apresentando perfis sigmoidais duplos para ambos os metais, apontando para diferentes possibilidades na estequiometria de formação destes complexos.Parâmetros farmacológicos calculados foram avaliados utilizando ferramentas disponíveis, apresentando bons resultados teóricos para atravessar barreiras biológicas. As propriedades de potencial eliminação de radicais foram estudadas com radicais superóxidos gerados eletroquimicamente, mostrando eliminação destes radicais com diferenças estatisticamente significativas nas ondas anódicas das espécies de superóxido estudadas.A interação destes ligantes com o peptídeo A(beta)(1-40) foi estimado através de experimentos por voltametria de onda quadrada, mostrando mudanças na corrente do voltamograma, sugerindo uma interação com este peptídeo. O comportamento assintótico da variação de corrente ao longo da adição do peptídeo A(beta)(1-40) indica a saturação na interação dos ligantes com o peptídeo. Como as placas de A(beta) constitui um dos mais importantes marcadores da DA, os sinais eletroquímicos gerados a partir da interação dos ligantes com este marcador demostra o potencial destes compostos como sondas eletroquímicas no contexto dessa doença. / [en] Alzheimer s Disease (AD) is the leading cause of dementia, with around 50 million patients worldwide in 2020, and this number is projected to triple by 2050, with an estimated cost of 1 trillion dollars annually. Biometals such as copper and zinc in their ionic forms can interact with the disease-related beta-amyloid peptide (Abeta), making it more susceptible to aggregation, in addition to contributing to increased oxidative stress in the body. In this context, ligands known as metallophores have been developed to sequester metal ions bound to the peptide in order to reduce A(beta) oligomerization and/or metal-induced oxidative stress. Instead of binding and systematically removing any metals from tissues, metallophores aim to act by restoring physiological metal homeostasis through specific binding with copper and/or zinc, attenuating their abnormal interactions with the peptide. In this scenario, two N-acylhydrazone ligands, with an electroactive fraction containing ferrocene, named Feizone and Ferfurone, were synthesized and characterized using techniques such as FTIR, (1)H NMR, TGA, UV-Vis, and XRD. The complexation of these ligands towards Cu(2+) and Zn(2+), as well as their amide deprotonation, were studied by cyclic voltammetry. The hydrolytic and photolytic stabilities were also assessed through electronic spectroscopy. Their radical scavenging potential was studied towards electrochemically generated superoxide ions and their suitability as electrochemical probes was estimated through interaction with the A(beta)(1-40) peptide by square-wave voltammetry. Both ligands demonstrated satisfactory stability compared to other molecules of the same class, also showing radical scavenging potential with statistically significant differences in the anodic waves of superoxide species. Additionally, experiments involving the full-length A(beta)(1-40) showed current changes in the voltammogram, suggesting their interaction with this peptide. Since plaques formed by A(beta) are one of the main hallmarks of AD, these ligands show potential as electrochemical probes in the context of the disease.