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Manejo de la producción de salinosporamide A en salinispora trópica CNB-440 empleando ingeniería metabólica y genéticaSaucedo Hernández, Vianey Diana January 2019 (has links)
Tesis para optar al grado de Doctora en Ciencias de la Ingeniería Mención Ingeniería Química y Biotecnología / Salinosporamide A is a cytotoxic that has been proven to combat various types of cancer and malaria. It is currently in phases II and III of approval as an anticarcinogen. The advantages that poses over other cytotoxics are greater activity at low concentrations and highly specific. It acts on the proteasome-ubiquitin system, responsible of apoptosis in cells.
This secondary metabolite is naturally ocurred in the actinomycetes bacterium strictly marine, Salinispora tropica that needs a specific ionic force in the medium to grow. Due to his nature it is a promisory source of secondary metabolites for pharmaceutical use hereby is constantly studied. The CNB440 strain is the representative strain of the species and posses a Genome-Scale Metabolic Model (GSM). The goal of this work was to implement diverse metabolic and genetic strategies that allow improve the production of Salinosporamide A.
Chapter 4 of this thesis details the proves to establish the protocols for growth and determination of Salinosporamide A, Define the sensitivity of bacteria to kanamycin (100 ug/ml), thiostreptone (12 µg/ml) and apramycin (12 µg/ml). The growth curves in several minimum mediums, and stablish the methodology for the determination of Salinosporamide A.
Chapter 5 describes the genetic strategy used to modify the bacterium and generate a higher concentration of Salinosporamide A. The strategy followed was by recombination homologous with the temperature sensitive vector (pGM1190) and transferred to S. tropica by conjugation with the strain E. coli ET12567/pUZ8002, to delete specific sites on the chromosome of S. tropica. These molecular tools have been successfully used in the transformation of various Streptomyces, but had not been tested in Salinispora. The sites suggested to be deletedto increase the production of the secondary metabolite were 3 clusters of genes sporolides, lymphostine and salinilactam. But due to various complications in the development of the present work only the deletion of the sporolide gene cluster was evaluated and this resulted in an increase of 20% in metabolite production.
Chapter 6 details the use of genome-scale metabolic model iCC908 for increase the production of Salinosporamide A. The first stage consisted in establishing the working environment of the model, to increase the accuracy of the model, integrated growth data, metabolites in medium production and determination of Salinosporamide A, with this was also able define in silico the supplementation of medium production, to obtain more Salinosporamide A. . The second stage consisted of applying different algorithms OptKnock, OptGene, OptOrf, GDLS, FSEOF, which browse reactions or genes within the genome-scale model that could be, deleted, blocked or overexpressed to increase the production of the secondary metabolite. We found several candidates that were evaluated in silico and we proposed to evaluate the deletion of two genes. As the last stage, were evaluated the metabolic pathways that increase production by gene overexpression. The evaluation of this metabolic pathways consist in add diverse substrates that increase the flow in the pathway of the gene to be overexpresed, tyrosine at a concentration of 5mM increase the production of the secondary metabolite Salinosporamide A by 180%, enhancing the presence of phenylalanine in the medium.
With these results it was possible to obtain a medium production that increased in 2.8 times Salinosporamide A by fermentation based on the use of the genome-scale metabolic model. And also was possible transform the strain S. tropica with genetic tools previously proved in Streptomyces.
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Fracture toughness and fracture surface energy of inorganic and non-metallic glasses / Ténacité et énergie de surface de fracture de verres inorganiques et non métalliquesTo, Theany 27 February 2019 (has links)
La ténacité et l’énergie de surface de fracture de verres, de vitrocéramiques et de composites à matrice en verre ont été étudiées. Tout d'abord, un test de flexion bi-axiale (configuration anneau/anneau) a été réalisé sur des verres de silice et des verre-à-vitres avec différentes conditions de surface afin d’identifier la relation entre le défaut de surface, la résistance et la ténacité du verre. Ensuite, trois méthodes d’expérimentation ont été mises en œuvre, principalement la méthode de poutre de flexion à pré-entaille droit (SEPB), la méthode de la poutre entaillée en chevron (CNB) et la méthode de la poutre avec indentations Vickers (VIF), afin de déterminer la ténacité de quatre verres produits industriellement et de déterminer les avantages et les inconvénients des différents méthodes sélectionnées. La méthode qui est apparue la plus fiable et auto-cohérente, la méthode SEPB (Single Edge Precrack Beam), a été appliquée à la détermination de la ténacité de nombreux verres et vitrocéramiques, afin d’étudier l’influence de la composition et de la microstructure sur les caractéristiques de fissuration (KIC et énergie de fissuration, γ). Enfin, l’influence de la température et de l'environnement sur la ténacité a été étudiée à l'aide de la méthode SEPB. Deux verres d'oxyde ont été testés à des températures élevées et avec une vitesse de charge de 10 MPa∙√m/s, une température de transition de 1,11Tg a été trouvée. Quatre autres verres d'oxyde ont été testés en environnement inerte et les mêmes valeurs de ténacité ont été obtenues à partir de deux vitesses de charge (100 fois) différentes. / Fracture toughness and fracture surface energy of commercial and laboratory glasses, glass-ceramics and glass matrix composites have been studied. First, bi-axial bending test (RoR configuration) was performed on fused silica and window float glasses with different surface conditions to identify the relationship between the surface flaw, the strength and fracture toughness. After, three experiment methods, mainly single-edge precracked beam (SEPB), chevron-notched beam (CNB) and Vickers indentation fracture (VIF) were performed to determine the fracture toughness of four commercial known glasses and to determine the advantages and inconveniences of the different selected methods. The method that is appeared as the most reliable and self-consistent, the SEPB (Single Edge Precrack Beam) method, was applied to determine the toughness of the large amount of glasses and glass-ceramics, to study the influence of the composition and the microstructure on the characteristics of cracking (KIC and fracture energy, γ). Last but not least, the influence of the temperature and environment on the fracture toughness was studied by means of the SEPB method. Two oxide glasses were tested in elevated temperatures and with the loading rate of 10 MPa∙√m/s, a transition temperature of 1.11Tg was found. Four other oxide glasses were tested in the inert environment and the same fracture toughness values were obtained from (100 times) two different cross-head speeds.
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Deset let fungování eurozóny - možné efekty pro nové členy včetně České republiky / Ten years of eurozone performance – potential effects for newcommers including Czech republicKnotek, Pavel January 2009 (has links)
The master thesis deals with european monetary integration. Its aim is to evaluate results of eurozone performance mainly for the first ten years of its existence. The thesis starts with short historical introduction, than continues by analysis OCA theory and Maastricht convergence criteria. The evaluation of effects, which eurozone brought to its members or candidates is made firstly from the point of view of monetary policy and than by confirming or refusing arguments for euro (non)adoption.
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Právní aspekty postavení a činnosti ČNB / Legal aspects of the status and activities of the Czech National BankRůžička, Roman January 2011 (has links)
The aim of this thesis is to describe and analyse the law position and activities of Czech national bank. The content of thesis is composed of three imaginary parts. In first one the historical development of central banking on Czech territory is described. It is devided into four chapters respecting the chronological development. The thesis is focused on a state of current legislation mainly, therefore the second and third part creates a widest portion of the thesis. The second part examines a framework of constitutional relations of Czech national bank and president, Parliament and goverment particularly, and the independency and aim of CNB in 7 chapters. The third part concerns with statutory law and is composed of 7 chapters also. It deals with a variety of activities of Czech national bank. Introduction, conclusion, abstract, key words and source list are drawn in other parts.
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Interações físicas entre o estuário do Rio Pará e a Plataforma Continental no norte do BrasilPRESTES, Yuri Onça 19 February 2016 (has links)
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Previous issue date: 2016-02-19 / Este trabalho tem foco em processos físicos oceanográficos que dominam a interação
entre o estuário do Rio Pará e a Plataforma Continental Amazônica (PCA). Esta,
amplamente estudada pela comunidade científica por diversas áreas do conhecimento,
comumente atrelada à influência do Rio Amazonas sobre a plataforma continental e/ou
às correntes oceânicas de contorno oeste que atuam nessa região do Oceano Atlântico
tropical. Contudo, uma importante questão ainda não respondida é qual a influência do
estuário do Rio Pará sobre a PCA? O objetivo do presente trabalho é analisar a interação
das forçantes físicas dentro do sistema Rio Pará-PCA. Medições eulerianas na
plataforma e no estuário foram realizadas de modo a abranger as variabilidades
temporais da área de estudo. Na plataforma, a coleta de dados ocorreu de Abril à Julho
de 2008, período que abrange a diminuição da descarga fluvial, mudanças na direção
dos alísios e intensificação da Corrente Norte do Brasil (CNB). No estuário, os
levantamentos realizados ocorreram ao longo de um ciclo completo de maré
semidiurno, durante o período seco (Outubro 2014) e o período chuvoso (Junho 2015)
da região amazônica. Também foi elaborado um modelo em caixa (box model) de
descarga para estivar a magnitude do aporte fluvial do Rio Pará. A propagação da onda
de maré domina o sistema, gerando fluxos bidirecionais em regiões de micromaré a
mais de 300 km de distância da foz do estuário. Na PCA, a maré gera intensas correntes
barotrópicas no sentido da componente de velocidade transversal à plataforma, até
alcançar a foz do sistema estuarino. Na quebra da plataforma, no período em que a CNB
se intensifica, suas bordas de corrente atingem a PCA em profundidades < 100 m,
agindo como um importante mecanismo nas trocas entre a plataforma amazônica e o
oceano profundo do Atlântico Tropical. A magnitude da descarga média estimada pelo
box model foi de 2 x 104 m3s-1, demonstrando também que o maior contribuinte hídrico
para o Rio Pará é o Rio Tocantins. A força de empuxo gerada pela descarga fluvial
interage com a força da maré, produzindo variabilidade lateral no estuário do Rio Pará.
Na margem direita, há tendência de fluxos laminares, estratificação vertical e correntes
baroclínicas, com processos mistura principalmente por difusão. Entre tanto, na margem
esquerda, foi verificado a presença de fluxos turbulentos, ausência de gradientes de
densidade vertical e correntes barotrópicas, de modo que os processos de mistura por
advecção dominam a mistura no sistema. Os resultados apresentados ao longo de três
artigos científicos enfatizaram o quão complexo e dinâmico é o sistema Rio Pará-PCA.
Esta característica dinâmica do sistema gera a extrema necessidade de que as atividades
potencialmente danosas devem ser desenvolvidas juntamente com medidas de
prevenção para evitar danos ao meio ambiente. / This work aims to study on oceanographic physical process that dominate the
interaction between the Pará River estuary and Amazon Continental Shelf (ACS). This
widely studied by the scientific community in various areas of geosciences, commonly
associated to Amazon River influence on the continental shelf, or to boundary ocean
currents which occur in western of Atlantic Tropical. However, an important question
not answered yet, is the influence of Para River estuary on the ACS? The objective of
this study is to analyze the interaction of physical forcings within the Pará River-ACS
system. Eulerian measurements on the platform and in the estuary were carried out to
cover the temporal variability of the study area. On the amazon shelf, data collection
occurred from period of April to July 2008 covering the decrease in river discharge,
changes in the direction of the trade and intensification of the North Brazil Current
(NBC). In the estuary, the surveys occur over a full semidiurnal tidal cycle during the
amazonian dry season (October 2014) and the rainy season (June 2015). It was also
prepared a discharge box model to estimate the magnitude of fluvial contribution of
Pará River on the ACS. The tidal wave propagation dominates the system, generating
bidirecional flows in microtides regions more than 300 km away from the estuary
mouth. In the ACS, the tides creates intense barotropic currents towards the across-shelf
velocity component until reach the mouth of the estuary system. On the shelf break, in
the period in which the CNB intensifies, its current edges reach the ACS at < 100 depth,
acting as an important mechanism in the exchange process between the Amazon shelf
and the deep ocean of the Atlantic Tropical. The estimated of average discharge
magnitude by box model was 2 x 104 m3s-1, also showing that the largest input to the
Pará River is the Tocantins River. The buoyancy force generated by river discharge
interacts with the tidal force, producing lateral variability in the estuary of Para River.
On the right margin, there is a tendency of laminar flow, vertical stratification and
baroclinic currents, with mixing processes primarily forced by diffusion. Meanwhile,
the left margin, it was verified the presence of turbulent flow, no vertical density
gradients, and intense barotropic currents, so that the advection processes dominate the
mixing in the system. The results presented over the three papers emphasize how
complex and dynamic is the Pará river-ACS system. This dynamic characteristic
generates extreme need for potentially damaging activities should be developed together
with preventive measures to avoid damage to the environment.
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Změny bankovní regulace a dohledu v EU v reakci na finanční krizi / Reaction to financial crisis: changes in bank regulation and supervision in the European UnionAndr, Jakub January 2010 (has links)
This thesis presents changes proposed in the European Union as a reaction to financial crisis. The aim of this thesis is to provide a summary and comment of changes both in organisation of supervision and banking regulation, especially concentrating on Basel III and taxation. The first part contains a description of the structure of supervision both in the Czech Republic and in the European Union. The description includes development in the Czech Republic in last five years. The second part contains a description of causes and development of the global financial crisis, defines systemic risk and financial stability and the ways to measure it. The third part describes new structure of supervision in the EU and impact of the change. It also includes a description of Basel III and CRD IV and its impact, discussion of options of taxation in the financial market. The end of this part is a brief summary of changes applied in the USA and its comparison to changes in the EU. The last chapter contains the CNB's position to these changes.
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Právní a ekonomické aspekty institutu věřitele poslední instance / Legal and economic aspects of the institute of the lender of last resortVágnerová, Tereza January 2021 (has links)
Legal and economic aspects of the institute of the lender of last resort Abstract The master thesis deals with the legal and economic aspects of the institute of the lender of last resort, as a key institute that helps ensure the stability of financial markets. The thesis is divided into four parts. The first part is focused on the economic aspects of this institute, especially deals with the general definition of the term lender of last resort and its historical genesis, as well as the principles and forms of assistance of the lender of last resort. The key knowledge of this part is the Thornton-Bagehot doctrine of the lender of last resort. The second part of the thesis deals with the legal regulation of the lender of last resort in the Czech Republic. The third part is aimed at relevant European legislation and provides a summary of the relevant case law of the Court of Justice of the European Union, namely the cases Pringle, Gauweiler and Weiss. The fourth and final part of the thesis outlines the legal framework of this institute in the USA. The second to fourth part of the thesis also analyzes the behavior of 3 central banks (CNB, ECB and FED) in fulfilling the role of lender of last resort during the global financial crisis of 2008, the European debt crisis and the current coronavirus crisis,...
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Dohled nad finanční činností / Supervision over financial activityDrahota, Jiří January 2017 (has links)
The aim of this diploma thesis is to describe and analyze supervision of financial activities in the Czech Republic. The thesis is structured into three chapters, which focuses on analysis of actual acts of CNB supervision in connection to the legal basis. The first chapter defines supervision of financial activities reflecting its historical development, economical aspect and basic trends in financial market regulation. The second chapter is the main part which describes the status and activities of the Czech National Bank in the position of the supervisory authority. The emphasis is placed especially on supervisory and regulatory activity, which represents the core of this thesis. Supervision is characterized from the methodological point of CNB's approach. Different types of supervision and regulation executions are also analyzed in this thesis.
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Česká národní banka a národní plán zavedení eura v České republice / The Czech National Bank and the national plan of adopting euro in the Czech RepublicTabačák, Anton January 2011 (has links)
- - 157 - - RESUMÉ or SUMMARY Keywords: Czech National Bank, National Changeover Plan, relationship, commercial banks, banking sector, financial market, europe structure of the banking sector, communication, responsibility of the CNB, non-financial sector, consumer protection, legislation - responsibility of the Ministry of Justice My dissertation thesis starts from the analysis of czech central banking, its history and role in modern europe economy. It refers to old famous,, historiae magistra vitae", because only problems, which had central banking in history, created central banking today. Special chapter is devoted to the Czech National Bank. The Czech National Bank is the central bank of the Czech Republic and the supervisor of the Czech financial market. It is established under the Constitution of the Czech Republic and carries out its activities in compliance with Act No. 6/1993 Coll., on the Czech National Bank and other regulations. It manages the assets entrusted to it by the state independently and with due diligence. Interventions in its activities are only permissible on the basis of a law. The supreme governing body of the CNB is the Bank Board, consisting of the CNB Governor, two Vice- Governors and four Chief Executive Directors. All Bank Board members are appointed by the President of the...
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Česká národní banka a národní plán zavedení měny euro v České republice / The Czech National Bank and the national plan of adopting euro in the Czech RepublicTabačák, Anton January 2012 (has links)
- - 179 - - RESUMÉ or SUMMARY Keywords: Czech National Bank, National Changeover Plan, relationship, commercial banks, banking sector, financial market, europe structure of the banking sector, communication, responsibility of the CNB, non-financial sector, consumer protection, legislation - responsibility of the Ministry of Justice My dissertation thesis starts from the analysis of czech central banking, its history and role in modern europe economy. It refers to old famous,, historiae magistra vitae", because only problems, which had central banking in history, created central banking today. Special chapter is devoted to the Czech National Bank. The Czech National Bank is the central bank of the Czech Republic and the supervisor of the Czech financial market. It is established under the Constitution of the Czech Republic and carries out its activities in compliance with Act No. 6/1993 Coll., on the Czech National Bank and other regulations. It manages the assets entrusted to it by the state independently and with due diligence. Interventions in its activities are only permissible on the basis of a law. The supreme governing body of the Czech National Bank is the Bank Board, consisting of the Czech National Bank Governor, two Vice-Governors and four Chief Executive Directors. All Bank Board members are...
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