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51	Využití molekulárně biologických technik pro identifikaci a analýzu probiotických bakterií / Use of Molecular Biology Techniques for Identification and Analysis of Probiotic Bacteria Konečná, Jana January 2019 (has links) Isolation of deoxyribonucleic acid (DNA) is an important step in the molecular diagnostics of microorganisms. A high quality of isolated DNA is necessary for DNA amplification by the polymerase chain reaction (PCR). The conventional DNA isolation using phenol-chloroform extraction and DNA precipitation in ethanol is time-consuming and requires the use of toxic phenol. Alternative method of DNA isolation is use of commercially available kits which, however, are expensive and their efficiency is low. Magnetic separation techniques using magnetic solid particles are one of modern methods to speed up the nucleic acids isolation. The aim of this work was to use two different types of magnetic particles for solid-phase DNA extraction. Magnetic microparticles P(HEMA – co – GMA) containing –NH2 group and nanoparticles PLL, whitch contains polylysine. The amounts of DNA in separation mixtures were measured using ultraviolet spectrophotometry (UV). The first experimental conditions were tested on chicken erythrocytes DNA. Phosphate buffer (pH 7, 7.6 and 8) was used for adsorption of DNA on magnetic particles. It was shown that approximately almost one half of DNA was adsorbed on the particles. The elution conditions of DNA were also optimized. Secondly, bacterial DNA was tested. After optimalization, the developed method was used for DNA isolation from real food supplements. This DNA eluted from the particles was in PCR ready quality. High resolution melting (HRM) curve analysis is a simple, low-cost method for amplicon discrimination and easy connection with real-time polymerase chain reaction (PCR). In this thesis, we report rapid species identification of strains belonging to the Lactobacillus group using HRM-PCR. Three different DNA isolation methods were used in this work: phenol extraction, separation using magnetic particles and commercial kit. Ten sets of targeted gene fragments primers (LAC1 – LAC2, LAC2 – LAC4, P1V1 – P2V1, Gro F – Gro R, 3BA-338f – Primer 1, V1F – V1R, CHAU - V3F – CHAU - V3R, CHAU - V6F – CHAU - V6R, poxcDNAFw – poxPromRVC, poxcDNAFw – poxPromRVT) were tested for amplification of the 16S rRNA gene. Use of GroF/R and LAC2/4 primers pairs successfully identify strains belong to the Lactobacillus group. The variance between used extraction methods for evidence of HRM curves was found.
52	Voltametrické studium interakce genotoxického 2-nitrofluorenu s DNA na visící rtuťové kapkové elektrodě / Voltammetric Study of the Interaction of Genotoxic 2-Nitrofluorene with DNA at a Hanging Mercury Drop Electrode Krejčová, Zuzana January 2011 (has links) In this Diploma Thesis, an interaction of genotoxic environmental pollutant 2-nitrofluorene with a double-stranded calf thymus DNA has been studied using a hanging mercury drop electrode (HMDE) as an electrochemical sensor. Two types of DNA damage were investigated and electrochemically detected (using cyclic voltammetry and differential pulse voltammetry): (i) The DNA damage caused by the direct interaction with 2-nitrofluorene and (ii) the DNA damage caused by short-lived radicals generated by the electrochemical reduction of the nitro group in 2-nitrofluorene. For the study of direct interaction, HMDE was modified by DNA and the interaction of DNA with 2-nitrofluorene was studied, after their incubation, right at the HMDE surface (adsorptive transfer stripping technique) or the DNA was preincubated with 2-nitrofluorene and, subsequently, the interaction was studied voltammetrically (DNA titration technique). Using both detection techniques, the formation of DNA - 2-nitrofluorene complex was observed and the mutual interaction was interpreted as an intercalation between the DNA base pairs, although such interaction was not clearly confirmed by UV-visible absorption spectroscopy. An electrostatic binding of 2-nitrofluorene on DNA sugar-phosphate backbone was partially formed at low concentrations of...
53	An evaluation of the management of deoxyrinucleic acid (DNA) evidence / An evaluation of the management of deoxyribonucleic acid (DNA) evidence Dywaba, Zukiswa Morencia 09 1900 (has links) DNA is identified as a powerful tool in the solving of rape cases, but it is often destroyed either by members of the public or the police officials who attend to the scene. The aim of the study was to evaluate the management of DNA evidence in rape cases in the Bishop Lavis Policing Area. To address the research topic under investigation, research questions, a legal framework and policies were used. The outcome of the study indicated poor performance in securing the crime scene and ensuring that physical evidence is preserved and not tampered with. On this basis, it was recommended that developmental workshops and intensive training on the management of DNA evidence be conducted to all members of the South African Police Service attend to rape crime scenes. This should be done to equip them with knowledge and an understanding of the management of DNA evidence. / Police Practice / M. Tech. (Forensic Investigation) Crime investigation Crime scene Deoxyribonucleic acid Forensic investigation Chain of custody Rape Individualisation Identification Physical evidence Management 363.25953209687355
54	Intégration d’une méthode d’actuation électrocinétique sur biocapteur plasmonique / Integrating an electrokinetic actuation method on a plasmonic biosensor Avenas, Quentin 20 December 2018 (has links) Cette thèse porte sur le développement d’un capteur plasmonique intégrant une fonction d’actuation des objets visés. L’objectif est de passer outre la limite de diffusion rencontrée à basse concentration en piégeant les particules sur la surface de détection. La stratégie adoptée est de structurer le film d’or servant à la détection de manière à pouvoir l’utiliser pour mettre en mouvement le fluide et les molécules par le biais de champs électriques. Le transfert de masse est réalisé par diélectrophorèse et électroosmose, deux effets électrocinétiques mis en oeuvre par des électrodes servant à la fois d’actuateur et de capteur plasmonique. Un état de l’art exhaustif et des simulations multiphysiques ont permis de concevoir un prototype de capteur intégré constitué d’électrodes interdigitées en or permettant la détection plasmonique. Le dispositif proposé a été obtenu par microfabrication en salle blanche puis caractérisé avant l’étude de ses performances. Une première phase de tests sur un système modèle, des billes de polystyrène dans de l’eau, a permis d’apporter la preuve de concept du fonctionnement du capteur, qui est effectivement capable de piéger rapidement les objets visés à sa surface afin de les détecter. Les mécanismes de transfert de masse ont été expliqués et la preuve de l’amélioration de la limite de détection par un facteur supérieur à 100 a été apportée. Dans un second temps, les performances du capteur appliqué à des objets biologiques ont été évaluées. Celui-ci piège efficacement des levures et des protéines, mais aucune amélioration n’a été observée dans le cas de la détection spécifique de l’hybridation entre deux brins d’acide désoxyribonucléique (ADN). Les causes de ce résultat ont été discutées et comprises et deux solutions différentes ont été explorées : l’adaptation de la fréquence d’opération et l’optimisation de la géométrie des électrodes. Ainsi, cette étude a permis de souligner la problématique de la mise en oeuvre d’effets électrocinétiques dans des milieux biologiques et de réfléchir aux pistes pertinentes pour sa résolution. / This thesis focuses on the development of an integrated plasmonic sensor capable to perform mass transport on targeted objects. The goal is to overcome the diffusion limit by trapping particules directly on the sensing surface. The adopted strategy was to structure the gold layer used for plasmonic detection in order to use the sofabricated structures to set the fluid and the molecules in motion by applying electric fields in the fluid. The mass transfer is realized through dielectrophoresis and electroosmosis, those two electrokinetic effects being operated by electrodes acting as sensor and actuator at the same time. An exhaustive state of the art as well as multiphysical simulations allowed us for designing a prototype for an integrated sensor consisting in gold interdigitated electrodes enabling plasmoninc sensing. The proposed device was obtained through microfabrication in clean room facilities and was characterized before the study of its performances. A first sequence of tests on a model system – polystyrene microbeads in water – brought the proof of concept we needed to validate the correct operation of the sensor, which is indeed capable of quickly trapping targeted objects on its surface and detecting them. The mass transfer mechanisms were explained and we showed the enhancement of the limit of detection by a factor greater than 100. In a second phase, performances of the sensor applied to biological objects were evaluated. It can effectively trap yeasts and proteins but no enhancement has been observed while detecting DNA hybridization events. Causes for this result were discussed and understood and two different solutions were explored: the adaptation of the operating frequency and the optimization of the electrodes geometry. Thus, this study highlighted the problematic of operating electrokinetic effects in biological media and suggested relevant leads towards its resolution. Electrotechnique Biocapteur Résonance de plasmon de surface - SPR Diélectrophorèse - DEP Electro-Osmose - ACEO Limite de détection Transfert de masse Electrodes interdigitées Acide désoxyribonucléique - ADN Electrotechnics BioSensor Surface Plasmon Resonance - SPR Dilectrophoresis - DEP Electro-Osmosis - ACEO Limit of detection Mass transfer Interdigitated electrodes Deoxyribonucleic acid - DNA 621.381 548 072
55	Dynamics and thermal behaviour of films of oriented DNA fibres investigated using neutron scattering and calorimetry techniques Valle Orero, Jessica 26 June 2012 (has links) (PDF) The majority of structural studies on DNA have been carried out using fibre diffraction, while studies of its dynamics and thermal behaviour have been mainly performed in solution. When the DNA double helix is heated, it exhibits local separation of the two strands that grow in size with temperature and lead to their complete separation. This work has investigated various aspects of this phenomenon. The experiments reported in this thesis were carried out on films of oriented fibres of DNA prepared with the Wet Spinning Apparatus. Thus, sample preparation and characterisation are essential parts of the research. The structures of two forms of DNA, A and B, have been explored as a function of relative humidity at fixed ionic conditions. A method to eliminate traces of ever-present B-form contamination in A-form samples was established. The high orientation of the DNA molecules within the samples allowed us to investigate dynamical fluctuations and the melting transition of DNA using neutron scattering, which can provide the spatial information crucial to understand a phase transition, probing the static correlation length along the molecule as a function of temperature. The transition has been investigated for A and B-forms in order to understand its dependence on molecular configuration.Furthermore, after the first melting, denatured DNA films show typical glass behaviour. Their thermal relaxation has been explored using calorimetry.Neutron and X-ray inelastic scattering (INS and IXS) were used in the past to measure longitudinal phonons in fibre DNA, and the results shown disagreement. Recent INS measurements supported with phonon simulations have been crucial to understand the different dispersion curves reported to date. Experiments using INS and IXS have been carried out to continue with this investigation. Attempts to observe the transverse fluctuations associated to the thermal denaturing of DNA, never experimentally investigated before, have been made. Deoxyribonucleic Acid Oriented DNA Fibres Wet Spinning Apparatus A-DNA B-DNA 1-D crystal X-ray fibre diffraction Neutron diffraction Inelastic neutron scattering Inelastic X-ray Scattering Differential scanning calorimetry Optic microscopy Thermal denaturation transition PBD model Enthalpy relaxation Structural relaxation Glass transition Phonon dispersion curves
56	Dynamics and thermal behaviour of films of oriented DNA fibres investigated using neutron scattering and calorimetry techniques / Dynamique et comportement thermique des films de fibres orientées d'ADN étudiés par les techniques de diffusion de neutrons et calorimétrie Valle Orero, Jessica 26 June 2012 (has links) La majorité des études structurales sur l’ADN avaient été réalisées par diffraction sur des fibres tandis que ses propriétés dynamiques thermiques avaient été étudiées en solution. Lorsque la double hélice d’ADN est chauffée elle présente des séparations locales des deux brins, dont la taille augmente avec la température jusqu’à la séparation complète des brins. Ce travail étudie différents aspects de ce phénomène. Les expériences présentées dans cette thèse ont été réalisées sur des films formés de fibres orientées d’ADN préparés par la méthode du ”filage humide“. La préparation et la caractérisation des échantillons en deux formes A et B de l’ADN ont constitué une partie importante de la recherche. Une méthode pour éliminer la contamination résiduelle de la forme B dans les échantillons de forme A a été mise au point. La bonne orientation des molécules d’ADN dans les échantillons nous a permis d’étudier les fluctuations dynamiques et la transition de dénaturation thermique de l’ADN par diffraction de neutrons, sensibles à la longueur de corrélation statique le long de la molécule en fonction de la température. La transition a été étudiée pour les formes A et B pour déterminer comment elle dépend de la conformation. De plus, après la première dénaturation thermique, les films d’ADN présentent un comportement typique d’un verre. Leur relaxation thermique a été étudiée par calorimétrie. La diffusion inélastique de neutrons et de rayons X (INS et IXS) avaient été utilisées antérieurement pour mesurer les phonons longitudinaux dans des fibres d’ADN, avec des désaccords entre les résultats. Des mesures INS récentes, complétées par des simulations, avaient été cruciales pour comprendre les différentes courbes de dispersion observées. Nous avons mené des expériences INS et IXS pour poursuivre cette analyse. Des tentatives pour observer les mouvements transversaux associés à la dénaturation thermique de l’ADN, jamais observés expérimentalement, ont également été faites. / The majority of structural studies on DNA have been carried out using fibre diffraction, while studies of its dynamics and thermal behaviour have been mainly performed in solution. When the DNA double helix is heated, it exhibits local separation of the two strands that grow in size with temperature and lead to their complete separation. This work has investigated various aspects of this phenomenon. The experiments reported in this thesis were carried out on films of oriented fibres of DNA prepared with the Wet Spinning Apparatus. Thus, sample preparation and characterisation are essential parts of the research. The structures of two forms of DNA, A and B, have been explored as a function of relative humidity at fixed ionic conditions. A method to eliminate traces of ever-present B-form contamination in A-form samples was established. The high orientation of the DNA molecules within the samples allowed us to investigate dynamical fluctuations and the melting transition of DNA using neutron scattering, which can provide the spatial information crucial to understand a phase transition, probing the static correlation length along the molecule as a function of temperature. The transition has been investigated for A and B-forms in order to understand its dependence on molecular configuration.Furthermore, after the first melting, denatured DNA films show typical glass behaviour. Their thermal relaxation has been explored using calorimetry.Neutron and X-ray inelastic scattering (INS and IXS) were used in the past to measure longitudinal phonons in fibre DNA, and the results shown disagreement. Recent INS measurements supported with phonon simulations have been crucial to understand the different dispersion curves reported to date. Experiments using INS and IXS have been carried out to continue with this investigation. Attempts to observe the transverse fluctuations associated to the thermal denaturing of DNA, never experimentally investigated before, have been made. Acide désoxyribonucléique Fibres orientées d’ADN Instrument de Filage Humide A-ADN B-ADN Cristal 1-D Diffraction de rayon X sur des fibres Diffraction de neutron Diffusion inélastique neutron Diffusion inélastique de rayon X Microscopie optique Transition de dénaturation thermique Modèle PBD Enthalpie de relaxation Relaxation structurale Transition vitreuse Courbe de dispersion de phonon Phonons longitudinaux et transverses Deoxyribonucleic Acid Oriented DNA Fibres Wet Spinning Apparatus A-DNA B-DNA 1-D crystal X-ray fibre diffraction Neutron diffraction Inelastic neutron scattering Inelastic X-ray Scattering Differential scanning calorimetry Optic microscopy Thermal denaturation transition PBD model Enthalpy relaxation Structural relaxation Glass transition Phonon dispersion curves
57	Jesus Christ’s humanity in the contexts of the pre-fall and post-fall natures of humanity: a comparative and critical evaluative study of the views of Jack Sequeira, Millard J. Erickson and Norman R. Gulley Mwale, Emanuel 12 1900 (has links) Bibliography: leaves 653-669 / Before God created human beings, He devised a plan to save them in case they sinned. In this plan, the second Person of the Godhead would become human. Thus, the incarnation of the second Person of the Godhead was solely for the purpose of saving fallen, sinful human beings. There would have been no incarnation if human beings had not sinned. Thus, the nature of the mission that necessitated the incarnation determined what kind of human nature Jesus was to assume. It was sin that necessitated the incarnation – sin as a tendency and sin as an act of disobedience. In His incarnational life and later through His death on Calvary’s cross, Jesus needed to deal with this dual problem of sin. In order for Him to achieve this, He needed to identify Himself with the fallen humanity in such a way that He would qualify to be the substitute for the fallen humanity. In His role as fallen humanity’s substitute, He would die vicariously and at the same time have sin as a tendency rendered impotent. Jesus needed to assume a human nature that would qualify Him to be an understanding and sympathetic High Priest. He needed to assume a nature that would qualify Him to be an example in overcoming temptation and suffering. Thus, in this study, after comparing and critically evaluating the Christological views of Jack Sequeira, Millard J. Erickson and Norman R. Gulley, I propose that Jesus assumed a unique post-fall (postlapsarian) human nature. He assumed the very nature that all human beings since humankind’s fall have, with its tendency or leaning towards sin. However, unlike other human beings, who are sinners by nature and need a saviour, Jesus was not a sinner. I contend that Jesus was unique because, first and foremost, He was conceived in Mary’s womb by the power of the Holy Spirit and was filled with the Holy Spirit throughout His earthly life. Second; He was the God-Man; and third, He lived a sinless life. This study contributes to literature on Christology, and uniquely to Christological dialogue between Evangelical and Seventh-day Adventist theologians. / Philosophy, Practical and Systematic Theology / D. Phil. (Systematic Theology) Adoptionism Alleles Anhypostatic Christology Anthropotokos Alternative Christology Apollinarianism Apthartodocetism Arianism Augustinian model Autosomal chromosomes Autosomal inheritance Born-again Christology Chalcedonian creed Christology from above Christotokos Chromosomes Chromosomal abnormalities Cloning Co-dominance Communicatio idiomatum Complementary base pairing Cri-du-chat syndrome Deoxyribonucleic acid (DNA) Diploid cell Docetism Dominant gene Down syndrome Dynamic incarnation Dynamic monarchianism Eastern Orthodox Christology Ebionitism Embryogenesis Eutychianism Evangelical Christology Existential Christology Functional Christology Genes Gene mutation Gene expression Genome Genotype Gnosticism Hamartiology Haploid cell Heterozygous History of Jesus’ Christology Homoiousios Theology Homozygous Human cloning Immaculate conception Incarnation Incarnational life Inheritance Karyotype Kenoticism Klinefelter syndrome Logos-flesh Christology Meiosis Mitosis Modalistic monarchianism Monarchianism Monophysitism Mutation Nestorianism Nicene Creed One-nature Christology Ontological Christology Organogenesis Patripassianism Phenotype Polygenic inheritance Post lapsarian view/model Prelapsarian view/model Recessive gene Roman Catholic Christology Sabellianism Seventh-day Adventist Christology Sex chromosomes Sex-linked inheritance Speculative Christology Theotokos Turner syndrome Two-natures Christology Unique post-fall Christology Socinianism Soteriology Theotokos Turner syndrome Two-natures Christology Unique post-fall Christology Virginal conception 231.4 Seventh-Day Adventists -- Doctrines Salvation -- Seventh-Day Adventists Fall of man -- History of doctrines Jesus Christ -- History of doctrines Sequerra, Jack Erickson, Millard J. Gulley, Norman R.

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