Made available in DSpace on 2016-06-02T19:30:11Z (GMT). No. of bitstreams: 1 TeseRDO.pdf: 756445 bytes, checksum: dd46098f781f2c54d8f4d7f9d1c6bb92 (MD5) Previous issue date: 2003-04-25 / Universidade Federal de Sao Carlos / Tropical fish generally inhabit environments with high temperatures and low dissolved oxygen. Along the evolutionary process several species developed mechanisms of air-breathing in order to compensate the oxygen demands caused by the aquatic hypoxia. These mechanisms required anatomical, physiological and biochemical adaptations. Studies on the effects of hypoxia and temperature changes on fish involve the comprehension of the cardio-respiratory mechanisms of compensation. The present study was addressed to determine the cardio-respiratory and air-breathing responses of jeju, Hoplerythrinus unitaeniatus, which utilize the swimming bladder as an organ for accessory respiration in the air, submitted to gradual hypoxia after acclimation to 15, 20, 25 and 30oC. The oxygen uptake ( &VO2 ), gill ventilation ( G V& ) breathing frequency (fR), ventilation volume (VT) and the O2 extraction from the ventilatory current (EO2) were obtained for specimens of jeju (Wt = 209,7 ± 5 g) by flow-trough respirometry. The air-breathing frequency (fRA) and time spent in air-breathing (TRA) were determined by means of an experimental chamber specially constructed for this purpose. Independently of the acclimation temperature, jeju responded to hypoxia as an oxyregulator, i. e., the species was able to keep constant &VO2 in response to graded hypoxia until reach a critical oxygen tension (PcO2). The mean &VO2 values at each acclimation temperature, before reaching the PcO2, were: 47 ± 0,8 mlO2.kg-1h-1 (15°C), 82 ± 0,3 mlO2.kg-1h-1 (20°C), 104 ± 2,6 mlO2.kg-1h-1 (25°C) and 112 mlO2.kg-1h-1 (30°C). The PcO2 for each acclimation temperature were 28, 33, 41 e 52 mmHg, respectively. The increased reductions on PcO2 as acclimation temperatures rose from 15 to 30oC showed that this species presents partial compensation (or type 3 compensation) to temperature increases. Jeju increased the G V& to compensate the graded hypoxia due to higher increments on VT than in fR. This kind of compensation, however, was not enough to keep a constant EO2, which decreased gradually in response to graded hypoxia in all acclimation temperature. At 25 and 30°C the fH were significantly higher than at 15 and 20oC. Hypoxic bradicardia was not recorded at 15 and 20oC and at 25 and 30°C it only occurred below the PcO2. The different PcO2 were also the threshold for the development of accessory air-breathing in all acclimation temperatures. Increases in both fRA and TRA were inversely proportional to the water PO2 reductions and directly proportional to the acclimation temperature. / Peixes de água doce tropical vivem em ambientes de altas temperaturas e baixas concentrações de O2. Isto resultou, ao longo do processo evolutivo, na necessidade da tomada do ar atmosférico em várias espécies de peixes, a fim de compensar seus requerimentos metabólicos, o que requereu modificações anatômicas, fisiológicas e bioquímicas. Estudos sobre o efeito da hipóxia e da variação da temperatura ambiental nos peixes devem envolver o conhecimento dos mecanismos de compensação cardiorespiratória. Assim, o objetivo deste estudo foi determinar as respostas cardiorespiratórios e da respiração aérea de jeju (Hoplerythrinus unitaeniatus), que utiliza a bexiga natatória como órgão acessório de troca gasosa, após período de aclimatação nas temperaturas de 15, 20, 25 e 30°C e submetido à hipóxia gradual. As medidas do consumo de O2 ( &VO2 ), ventilação branquial ( G V& ), freqüência respiratória (fR), volume ventilatório (VT) e extração de O2 da corrente ventilatória (EO2) foram obtidas de exemplares de jeju (Wt = 209,7 ± 5 g) por meio de respirometria de fluxo constante. A freqüência (fRA) e o tempo da respiração aérea (TRA) também foram determinados utilizando-se de uma câmara especialmente desenvolvida para esta finalidade. Independentemente da temperatura de aclimatação, H. unitaeniatus comportou-se como um oxi-regulador, mantendo uma &VO2 constante por amplos intervalos de reduções de O2 até atingir a tensão crítica de O2 (PcO2). Nas diferentes temperaturas a &VO2 foi de 47 ± 0,8 mlO2.kg-1h-1 a 15°C, 82 ± 0,3 mlO2.kg-1h-1 a 20°C, 104 ± 2,6 mlO2.kg-1h-1 a 25°C e 112 mlO2.kg-1h-1 a 30°C. As PcO2 nas respectivas temperaturas foram de 28, 33, 41 e 52 mmHg. A redução crescente das PcO2 mostrou que esta espécie apresenta uma compensação parcial ou do tipo 3 aos aumentos crescentes na temperaturas de aclimatação. O jeju aumentou a G V& em resposta à hipóxia gradual valendo-se de maiores incrementos no VT do que na fR. Entretanto, este tipo de compensação não foi suficiente para manter uma constante EO2, a qual decresceu gradualmente em função da hipóxia. A fH a 25 e 30°C foi significativamente mais elevada do que a 15 e 20°C. A bradicardia hipóxica só foi registrada nas temperaturas de 25 e 30°C e somente ocorreu em tensão abaixo das respectivas PcO2. As PcO2 foram o limiar para o início da respiração aérea acessória em todas as temperaturas estudadas. Os aumentos na fRA, assim como da TRA foram inversamente proporcionais às reduções da PO2 da água e diretamente proporcionais às temperaturas de aclimatação.

L'augmentation du pouvoir électrocatalytique d'une pile à combustible fonctionnant à haute température (SOFC, Solid Oxide Fuel Cell) passe notamment par l'amélioration des performances de la cathode. A cet effet, notre objectif a été de créer puis mesurer une conductivité mixte (électronique ou ionique) au sein de nouveaux oxydes de structure perovskite ABO3-d. Une étude fondamentale a été menée sur des composés choisis en fonction de l'état de valence et de la taille du cation B et du taux de lacunes d. La caractérisation des propriétés de transport (électronique et ionique) a été réalisée à l'aide de techniques nouvelles au laboratoire (meusure de la diffusion de 18O). Par la suite une corrélation entre les deux types de conductivité et les propriétés physico-chimiques des matériaux (paramètres de maille, polarisabilité des ions, ...) a été proposée.

The growing problem of antibiotic resistance and the lack of promising prospective antibiotics have forced us to search for new classes of antibiotics. Among the candidates to develop into future antibacterials are antimicrobial peptides (AMPs). These potent, broad spectrum compounds are important components of innate immunity of organism from all kingdoms of life. One such family of mini-proteins from plants is called cyclotides, whose members are defines by cyclic backbone and a cystine knot (CCK), which confers to them extreme stability in the face of biological, chemical and physical insults.     Some cyclotides possess Gram-negative specific antibacterial activity; the purpose of this thesis was to characterize how these molecules kill bacteria, and how bacteria would respond to treatment with cyclotides. For this purpose, Salmonella enterica and Escherichia coli mutants resistant to the cyclotides cycloviolacin O2 and cycloviolacin O19, respectively, were selected. These mutants were characterized by whole genome sequencing, genetic reconstitution, fitness measurements, and cross-resistance studies. These studies identified a number of genetic pathways for resistance development to cyclotides. These mutants displayed variable fitness profiles in laboratory growth media and in mice competition experiments, with some mutants possessing a fitness advantage in mice. Cross-resistance studies resulted in the identification of several cases of cross-resistance and collateral sensitivity between cyclotides and other AMPs/antibiotics.      Antimicrobial effects of cyclotides were assayed in different conditions and in bacterial organisms with different surface characteristics. In addition, immunolocalization experiments were performed to explore the biological distribution of cyclotides in plants and to determine the mechanism of action of cyclotides in bacteria, respectively. Antibodies raised against cyO2 were used for this purpose. Immunohistochemical techniques applied to plant cells, tissues and organs provided the information that cyclotides were distributed in all plant organs, and were found in tissues vulnerable to pathogen attack, and that cyclotides were stored in the vacuoles of plant cells. Immunogold staining of cyclotide treated cells of S. typhimurium, showed effects of cyclotide treatment on the cell envelope components as well as cytoplasm. A higher number of cyclotide molecules was associated with the cell envelope, but a considerable fraction of them penetrated into the cytoplasm.

L’endurance aérobie est rarement mesurée à des intensités maximales, mais plutôt sous-maximales, principalement via des protocoles à puissance ou à durée constante. Or, les conditions réelles d’une épreuve sportive en compétition sont mieux représentées par le maintien de la plus haute puissance métabolique possible, et ce, le plus longtemps possible. Cela implique une autogestion de la puissance de travail de la part du participant – qui peut donc varier tout le long de l’épreuve – ainsi que l’atteinte des capacités aérobies maximales, dans certains cas. Dans cette étude, nous avons voulu comparer les variations de certains paramètres ventilatoires (ventilation, volume courant, fréquence respiratoire) et systémiques (rendement mécanique externe brut [RMEC], consommation d’oxygène [VO2]) entre deux protocoles d’effort maximal, l’un à puissance constante (100 % PAM), l’autre à puissance décrémentée (> 95 % VO2max). En effet, comparativement à un protocole d’effort classique à puissance constante, un protocole décrémenté implique l’utilisation de VO2 et non de la puissance mécanique comme critère de sélection de l’intensité d’effort, et augmente la durée de l’épreuve. En conséquence, le maintien de la consommation d’oxygène maximale (VO2max), et non de la puissance aérobie maximale (PAM), nous a permis d’observer des réponses physiologiques différentes de celles généralement décrites lors d’efforts incrémentés ou à puissance constante. Ces différences se sont traduites par une fréquence respiratoire plus élevée et un volume courant plus bas, indiquant l’existence d’une réponse tachypnéique plus prononcée lors d’un protocole décrémenté par rapport à un protocole à puissance constante, et reflétant ainsi une respiration plus superficielle, probablement due à une fatigue plus accrue des muscles ventilatoires. Cette fatigue serait caractérisée par un rendement mécanique plus bas et un VO2 plus élevé, et ce, malgré la diminution progressive de la puissance de travail lors de l’atteinte d’une consommation d’oxygène proche de VO2max. / Aerobic endurance is more commonly measured at submaximal rather than maximal work intensities, mainly through constant-power output trials, or time trials. However, real competition conditions are better represented through the ability to sustain the highest power output (PO) for the largest period of time. That implies the use of self-pacing strategies, which leads power output to vary throughout the exercise duration, and can also induce the attainment of maximal oxygen consumption (VO2max), in some cases. The aim of this study was thus to compare the variation of some ventilatory (tidal volume, breathing frequency) and systemic (gross mechanical efficiency, O2 cost) parameters, between two maximal endurance protocols, the first one consisting of a constant-load test (100% MAP) and the second one a decremental test at near-maximal aerobic capacity (> 95 % VO2max). Indeed, compared to a “classical” constant-power output test, a decremental test implies the use of VO2 rather than mechanical PO to control intensity throughout the task, and a higher effort duration. Consequently, we observed that the sustainment of maximal O2 consumption rather than maximal aerobic power (MAP) led to different physiological responses from those usually observed during constant-power output tests. These differences were caracterized by a higher breathing frequency as well as a lower tidal volume during the decremental test, which both reflect a more exagerated tachypneic response, and therefore indicate the existence of a shallower breathing pattern in this type of effort, probably due to a higher ventilatory muscle fatigue. This fatigue could also be expressed through a lower gross mechanical efficiency and a higher O2 cost, despite a progressive decrease of mechanical power as soon as oxygen consumption gets near VO2max.

This paper is about usage of accounting information in financial analysis. In the first part financial statements under Czech Accounting Standards and under International Financial Reporting Standards are described. The most important differences are discussed. In the second part methods of financial analysis are explained. In the third part practical financial analyses are done for the companies Vodafone Czech Republic, a.s. (reports under CAS) and Telefónica O2 Czech Republic, a.s. (reports under IFRS). Main differences in financial analysis which rise from different accounting standards are discussed.

L’activité catalytique de l’or en réaction d’oxydation préférentielle du CO (PROX) dépend de plusieurs facteurs tels que la nature du support, l’ajout de promoteurs, la taille des particules ou la nature du prétraitement. L’objectif de cette thèse a donc été de clarifier l’influence de ces paramètres sur cette réaction en utilisant des catalyseurs à base d’or modifiés. Dans une 1ère partie, nous avons étudié un système à base d’or supporté sur des oxydes Ce-Fe préparés par co-imprégnation ou imprégnations successives en faisant varier le rapport Ce/Fe. Le dopage de la cérine par le fer conduit à la formation d’une solution solide qui améliore l’activité en PROX en raison d’une mobilité d’oxygène accrue. Dans une 2ème partie, nous avons évalué une série de supports (Al2O3, CeO2 et Ce2Zr2O7) pour optimiser le dépôt de Au, Cu ou Au-Cu. La combinaison Au-Cu/CeO2 s’est révélée être la meilleure en PROX. Nous avons donc étudié l’influence du rapport Au/Cu (1/3, 1/1 ou 3/1) ainsi que l’influence du pré-traitement (calcination ou réduction) sur ce système. Les catalyseurs ont été systématiquement caractérisés par des méthodes physico-chimiques (BET, DRX, MET, XPS, RTP, FTIR). Les performances catalytiques du système Au-Cu/CeO2 dépendent fortement de la taille des particules, de l’état chimique de l’or déterminé par XPS, de l’interaction Au-support et de la distribution de surface des 2 métaux. Une étude cinétique corrélée à de l’infrarouge à transformée de Fourier a permis d’établir un mécanisme de réaction d’oxydation de CO dans le lequel CO adsorbé réagit avec O2 adsorbé pour conduire à la formation de CO2. / The catalytic activity of gold for CO preferential oxidation in rich hydrogen (PROX) is generally influenced by various parameters such as the nature of support, the addition of a promoter, the particle size or the pre-treatment. Therefore, the objectives of this work were to clarify the influence of these parameters on a series of modified gold catalysts in PROX reaction. In a 1rst part, we have studied a system based on Au supported on Ce-Fe oxides prepared by co-precipitation or impregnation with various Ce/Fe ratios. The doping of CeO2 by iron led to the formation of a solid solution which improved the PROX activity due to an increase of the oxygen mobility. In a 2nd part, a series of supports (Al2O3, CeO2, and Ce2Zr2O7) was evaluated in order to load Au, Cu or Au-Cu. The combination of Au-Cu/CeO2 was shown to be the best system. The influence of Au/Cu (1/3, 1/1 or 3/1) ratio was therefore undertaken along with the effect of the pre-treatment (calcination or reduction). The catalysts have been systematically characterized by physico-chemical techniques (BET, XRD, TEM, XPS, TPR, FTIR). The kinetics of PROX reaction and CO oxidation mechanism on Au-Cu/CeO2 were explored and correlated with an FTIR study. The catalytic performances of these catalysts are strongly dependent upon the particle size of the metal, the chemical state of gold, the gold-support interaction and the surface distribution of Au and Cu. We have proposed a CO oxidation mechanism in which adsorbed CO reacts with adsorbed O2 to produce CO2.

The humidity, temperature, wind speed/direction micro sensors can be manufactured individually, resulting in three individual substrates. The integration of the three sensors into a single substrate is a vital challenge to achieve an integrated intelligent sensor so called a multiple sensor. This requires the integration of process flows and is a major challenge because adequate sensor performance must be maintained. Polyimide was selected as a humidity sensing material for its compatibility with conventional integrated circuit fabrication technology, negligible temperature dependence and good resistance against contamination. Nickel was selected for the temperature and wind speed/direction sensor because of its useful temperature coefficient and the advantage of its cost. Since the known wet etchant for nickel requires hard-baked photoresist, a method which does not attack the polyimide while removing the photoresist must be developed. The method developed for etching nickel employs hard-bake-free photoresist. Other challenge was ensuring good thermal isolation for the wind speed/direction sensor fabricated on a silicon nitride layer preformed on top of a silicon wafer. Since silicon acts as a good heat sink, the silicon under the sensor was etched entirely away until the silicon nitride layer was reached. This structure achieved good thermal isolation resulting in small power consumption. This low power feature is essential for sensors deployed in fields where power access or replacement of power sources is restricted. This structure was compared with the structure created by polyimide plateau on a silicon nitride layer coated on a silicon substrate as a function of power consumption. Based on the examination of thermal isolation, the multiple sensor utilizing a MEMS technique was fabricated with a single-sided mask aligner. The characteristics of humidity sensors fabricated with polyimide were examined in detail with respect to variations of electrode structures, improvement of sensitivity, effect of process temperature, temperature and frequency dependence, and stability. The humidity sensor constructed with O2 plasma treated polyimide resulted an improvement in sensitivity and hysteresis. The investigation using XPS, FTIR and AFM concluded the chemical modification of polyimide played an important role in this improvement. The design, fabrication and results of a series of humidity sensors are quantified. There is always no unique packaging solution for sensors because of the application-specific nature of the sensors. This intelligent environmental monitoring system was designed to accommodate both an environmental sensor and its signal conditioning electronics circuitry (SICONEC) into a single package. The environmental sensors need direct exposure to the environment while SICONEC needs a sealed encapsulation to avoid environmental damage. A new style of packaging addressing these requirements was demonstrated using a hot embossing machine. The hot embossing machine was used to embed an integrated circuit (IC) in a bare die condition into a polycarbonate (PC) sheet. In this case, the IC was flipped down against the PC, which protects the front side of the IC from the environmental damages. In a test phase, a die containing operational amplifiers was embossed into the PC. A humidity sensor and surface mount resisters were placed on the same surface of the PC to test the validity of this new technique. Interconnection between the embossed die and the humidity sensor was established using bonding wires. Copper tracks were also used to ensure all electrical connections for the die, the humidity sensor and the resistors. The results clarified the method developed. Details of process methods, issues and further potential improvement are reported.

<p>In the present thesis the density functional B3LYP has been used to study reactions of NO and O₂ in redox active enzymes.</p><p>Reduction of nitric oxide (NO) to nitrous oxide (N₂O) is an important part in the bacterial energy conservation (denitrification). The reduction of NO in three different bimetallic active sites leads to the formation of hyponitrous acid anhydride (N₂O₂^2-). The stability of this intermediate is crucial for the reaction rate. In the two diiron systems, respiratory and scavenging types of NOR, it is possible to cleave the N-O bond, forming N₂O, without any extra protons or electrons. In a heme-copper oxidase, on the other hand, both a proton and an electron are needed to form N₂O.</p><p>In addition to being an intermediate in the denitrification, NO is a toxic agent. Myoglobin in the oxy-form reacts with NO forming nitrate (NO₃ ^-) at a high rate, which should make this enzyme an efficient NO scavenger. Peroxynitrite (ONOO^-) is formed as a short-lived intermediate and isomerizes to nitrate through a radical reaction.</p><p>In the mechanism for pumping protons in cytochrome oxidase, thermodynamics, rather than structural changes, might guide protons to the heme propionate for further translocation.</p><p>The dioxygenation of arachidonic acid in prostaglandin endoperoxide H synthase forms the bicyclic prostaglandin G₂, through a cascade of radical reactions. The mechanism proposed by Hamberg and Samuelsson is energetically feasible.</p>

In the present thesis the density functional B3LYP has been used to study reactions of NO and O2 in redox active enzymes. Reduction of nitric oxide (NO) to nitrous oxide (N2O) is an important part in the bacterial energy conservation (denitrification). The reduction of NO in three different bimetallic active sites leads to the formation of hyponitrous acid anhydride (N2O22-). The stability of this intermediate is crucial for the reaction rate. In the two diiron systems, respiratory and scavenging types of NOR, it is possible to cleave the N-O bond, forming N2O, without any extra protons or electrons. In a heme-copper oxidase, on the other hand, both a proton and an electron are needed to form N2O. In addition to being an intermediate in the denitrification, NO is a toxic agent. Myoglobin in the oxy-form reacts with NO forming nitrate (NO3 -) at a high rate, which should make this enzyme an efficient NO scavenger. Peroxynitrite (ONOO-) is formed as a short-lived intermediate and isomerizes to nitrate through a radical reaction. In the mechanism for pumping protons in cytochrome oxidase, thermodynamics, rather than structural changes, might guide protons to the heme propionate for further translocation. The dioxygenation of arachidonic acid in prostaglandin endoperoxide H synthase forms the bicyclic prostaglandin G2, through a cascade of radical reactions. The mechanism proposed by Hamberg and Samuelsson is energetically feasible.

A novel process for the fabrication of Long Range Surface Plasmon Polariton (LRSPP) waveguide based biosensors is presented herein. The structure of the biosensor is comprised of Au stripe waveguide devices embedded in thick CYTOP claddings with a SiO2 solvent diffusion barrier and etch-stop layer. The SiO2 layer is introduced to improve the end quality of Au waveguide structures, which previously deformed during the deposit of the upper cladding process and to limit the over-etching of CYTOP to create micro-fluidic channels. The E-beam evaporation method is adapted to deposit a thin SiO2 on the bottom cladding of CYTOP. A new micro-fluidic design pattern is introduced. Micro-fluidic channels were created on selective Au waveguides through O2 plasma etching. The presented data and figures are refractive index measurements of different materials, thickness measurements, microscope images, and AFM images. Optical power cutback measurements were performed on fully CYTOP-cladded symmetric LRSPP waveguides. The end-fire coupling method was used to excite LRSPP modes with cleaved polarization maintaining (PM) fibre. The measured mode power attenuation (MPA) was 6.7 dB/mm after using index-matched liquid at input and output fibre-waveguide interfaces. The results were compared with the theoretical calculations and simulations. Poor coupling efficiency and scattering due to the SiO2 are suspected for off-target measurements.