In my studio practice I attempt to create sensual experiences from which nothing is learned and lack resolution; this thesis is an attempt to find ways to outline these experiences. However, this attempt will eventually lead to something entirely different, for these experiences are undefinable and impossible to investigate. I will try to show how the components of the work, rather than forming a direct link to a chain of thoughts or associations, summon an experience that is derived only from the encounter itself.

It is expected that the massive expansion of carriage of goods that went on in past couple of decades will soon resume after the worldwide recession. Due to its flexibility, speed, ability to deliver "door to door", low price and other positive qualities the most popular type of carriage is carriage of goods by road, which is also the type of carriage of goods that this thesis is mainly focused on. The goal of the thesis is to describe and explain legal aspects of international carriage of goods by road with a special focus on the liability of the carrier. The most important legal regulation that governs the area of international carriage of goods by road is the Convention on the Contract for the International Carriage of Goods by Road (hereinafter the CMR Convention) that has been signed on May 19, 1956. It is an exceptional piece of legislation, which is in effect for more then fifty years with only two minor changes since its adoption.

The purpose of this thesis was to analyse and compare isseu of carrier's liability in international carriage of goods. Thesis is devided in seven parts, which offers outline of carrier's liability in particular modes of carriage. First part deals with term contract for the carriage according civil code, parts of this contract as well as another contracts which aim is carriage. Furthermore deals with term liability and put outline of distinction among strict liability and liability for fault. Last subchapter of first part describes term carriage. Second part describes legal framework of contract for the international carriage of goods and specifies distinction among choice of law and direct method. Remaining parts refer to carrier's liability according international treaties concerned with particular modes of carriage, i. e. international carriage of goods by road, by rail, by air, by sea and by inland waterways. Bigger attention is dedicated to third part which deals with carrier's liability in international carriage of goods by road. Emphasis to this part is given, because international carriage of goods by road is the most frequently one - especially from Czech point of view. International carriage of goods by road is for sixty years ruled by CMR Convention which was amended only twice. This mode...

The present work focuses on two strategies contributing to the development of high efficiency, cost-effective photovoltaic (PV) technology for renewable energy generation: the design of new materials offering enhanced opto-electronic performance and the investigation of material degradation processes and their role in predicting the long-term reliability of PV modules in the field. The first portion of the present work investigates the integration of a novel CdTe-ZnO nanocomposite material as a spectral sensitizer component within a thin-film, hybrid heterojunction (HJ) PV device structure. Quantum-scale semiconductors have the potential to improve PV device performance through enhanced spectral absorption and photocarrier transport. This is realized via appropriate design of the semiconductor nanophase (providing tunable spectral absorption) and its spatial distribution within an electrically active matrix (providing long-range charge transport). Here, CdTe nanocrystals, embedded in an electrically active ZnO matrix, form a nanocomposite (NC) offering control of both spectral absorption and photocarrier transport behavior through the manipulation of nanophase assembly (ensemble effects). A sequential radio- frequency (RF) magnetron sputter deposition technique affords the control of semiconductor nanophase spatial distribution relative to the HJ plane in a hybrid, ZnO-P3HT test structure. Energy conversion performance (current density-voltage (J-V) and external quantum efficiency (EQE) response) was examined as a function of the location of the CdTe nanophase absorber region using both one dimensional solar cell capacitance simulator (SCAPS) and the experimental examination of analogous P3HT-ZnO based hybrid thin films. Enhancement in simulated EQE over a spectral range consistent with the absorption region of the CdTe nanophase (i.e. 400–475 nm) is confirmed in the experimental studies. Moreover, a trend of decreasing quantum efficiency in this spectral range with increasing separation between the CdTe nanophase region and the heterojunction plane is observed. The results are interpreted in terms of carrier scattering/recombination length mitigating the successful transport of carriers across the junction. The second portion of the research addresses the need for robust PV performance in commercial module as a primary contributor to cost-effective operation in both distributed systems and utility scale generation systems. The understanding of physical and chemical mechanisms resulting in the degradation of materials of construction used in PV modules is needed to understand the contribution of these processes to module integrity and performance loss with time under varied application environments. In this context, the second part of present study addresses microcracking in Si–an established degradation process contributing to PV module power loss. The study isolates microcrack propagation in single-crystal Si, and investigates the effect of local environment (temperature, humidity) on microcrack elongation under applied strains. An investigation of microindenter-induced crack evolution with independent variation of both temperature and vapor density was pursued in PV-grade Si wafers. Under static tensile strain conditions, an increase in sub-critical crack elongation with increasing atmospheric water content was observed. To provide further insight into the potential physical and chemical conditions at the microcrack tip, micro-Raman measurements were performed. Preliminary results confirm a spatial variation in the frequency of the primary Si vibrational resonance within the crack-tip region, associated with local stress state, whose magnitude is influenced by environmental conditions during the period of applied static strain. The experimental effort was paired with molecular dynamics (MD) investigations of microcrack evolution in single-crystal Si to furnish additional insight into mechanical contributions to crack elongation. The MD results demonstrate that crack-tip energetics and associated cracking crystal planes and morphology are intimately related to the crack and applied strain orientations with respect to the principal crystallographic axes. The resulting fracture surface energy and the stress-strain response of the Si under these conditions form the basis for preliminary micro-scale peridynamics (PD) simulations of microcrack development under constant applied strain. These efforts were integrated with the experimental results to further inform the mechanisms contributing to this important degradation mode in Si-based photovoltaics.

Zusammenfassungen 83 5 Zusammenfassungen 5.1 Zusammenfassung Durch Expressionsklonierung wurde 1994 der erste organische Kationentransporter, rOCT1, aus der Ratte isoliert (Gründemann et al., 1994). 1999 wurde eine Aminosäure in der 11. Transmembrandomäne von rOCT1 entdeckt, welche Teil der Substratbindungstasche dieses Transporters war (Gorboulev et al., 1999). Zielsetzung der vorliegenden Arbeit war es weitere funktionell relevante Aminosäuren zu identifizieren. Ein Vergleich der „Helical Wheels“ aller 12 hypothetischen Transmembrandomänen zeigte eine Akkumulation von 5 OCT und OCTN spezifischen Aminosäuren in der vierten Transmembrandomäne auf einer Seite. Bei diesen Aminosäuren handelte es sich um K215, W218, Y222, T226 und V229. Es wurden verschiedene Punktmutationen an diesen Positionen eingeführt. Es zeigte sich mit Hilfe radioaktiv markierter Substrate von rOCT1, dass selbst Substitutionen durch strukturell verwandte Aminosäuren bei den flankieren Aminosäuren zu einem Ausfall des rOCT1 vermittelten Substrattransport führte. Weiterhin schien an Position 218 für den rOCT1- vermittelten Transport von TEA eine aromatische Aminosäure von großer funktioneller Relevanz zu sein. Wir vermuten hier eine Kationen p-Elektronen Interaktion des aromatischen Ringsystems des Tryptophans mit der positiven Ladung des TEA. Versuche mit den Mutanten des Tyrosins 222 zeigten ebenfalls Änderungen bei den Transportraten und Affinitäten verschiedener Substrate. Eine Kationen-p Interaktion konnte ausgeschlossen werden, jedoch war die Affinität der Mutante Y222F zum TPeA um einen Faktor 20 gegenüber dem Wildtyp erhöht. Weiterführende Untersuchungen mit der Zwei- Elektrodenspannungsklemme zeigten unterschiedliche Affinitäten des TPeA zum Wildtyp im Vergleich zum mutierten Protein in seiner nach außen bzw. nach innen gerichteten Konformation. War die Form der Inhibierung des TEA-induzierten Stromes durch TPeA beim Wildtyp kompetitiv, so zeigte sie bei der Mutante einen nicht-kompetitiven Charakter. Die Mutante T226A zeigte ebenfalls Änderungen in Affinität und Selektivität. Bei allen transportierenden Mutanten zeigte sich, dass der Transport von MPP nicht oder kaum verändert war, hingegen wurden sehr starke Änderungen der Transportcharakteristika von TEA gefunden, was auf verschiedene Substratbindungsstellen in rOCT1 hinweist. Diese Versuche zeigen deutlich die funktionelle Relevanz und die Beteiligung der mutierten Aminosäurepositionen an der Substratbindetasche von rOCT1. In Versuchen, in welchen Chinin als Inhibitor des rOCT2 vermittelten Transports genutzt wurde, passierte Chinin mittels Diffusion in seiner ungeladenen Form die Oozytenmembran und hemmte rOCT2 von der Innenseite. Dies könnte der Grund für die nicht-kompetitive Form der Inhibition der TEA-Aufnahme durch Chinin sein. Diese Versuche wurden dadurch bestätigt, dass die protonierte Form des Chinins eine kompetitive Form der Inhibition zeigte und den Transporter von außen hemmte. / The first organic cation transporter was cloned from rat (rOCT1) in 1994 by using the expression cloning technique (Gründemann et al., 1994). In 1999 it was found that one of the aminoacids in the 11th transmembranedomain of rOCT1, is part of it’s substrate binding pocket (Gorboulev et al., 1999). It was the aim of this thesis to identify more aminoacids of functional relevance for this transport protein. A comparison of all 12 helical wheels of the transporter showed, that there is an accumulation of 5 OCT and OCTN specific aminoacids at one side of the a-helix. These aminoacids were K215, W218, Y222, T226 and V229. Different single point mutations have been generated at these positions. Using radiolabeled substrates for rOCT1, the experiments showed, that even a substitution by a structural related aminoacid at the flanking aminoacids resulted in a failure of substrate transport. For TEA transport it has been suggested that the aminoacid at position 218 should have aromatic properties. We therefor suggest a cation-p interaction between the aromatic ringsystem of W218 and the positive charge of TEA. Experiments with mutations in position Y222 also showed differences in transport rates and affinities referring to the wildtype using different substrates. A cation-p interaction could be excluded, but it was shown, that there was a 20fold increased affinity of TPeA for the mutant Y222F. Further experiments utilizing the two electrode voltage clamp technique showed different affinities for wildtype rOCT1 in comparison to the mutated protein in its outward and inward conformation. The wildtype showed a competitive type of inhibition for TPeA inhibited TEA current, the mutant showed a non-competitive type of inhibition. The mutant T226A also showed changes in affinity and selectivity. In all transporting mutants we found no or lowest changes in the transport characteristics of MPP, but very big changes in the transport characteristics of TEA, an indication for different binding sites for these substrates. The experiment clearly showed, that these five aminoacids are of functional relevance for rOCT1 transport. In experiments using quinine as inhibitor of rOCT2 mediated transport quinine in it’s uncharged form passed the oocyte membrane by diffusion and inhibited rOCT2 from the inside. This might be the reason for the non-competitive type of inhibition, using quinine as the inhibitor for TEA uptake. This hypothesis was confirmed when we showed that the type of inhibition changed into the competitive type, when we used the protonated form of quinine.

A bixina é o principal carotenoide encontrado na superfície externa das sementes de Bixa orellana L., conhecida popularmente como urucum. Os extratos das sementes são largamente utilizados, tradicionalmente como condimento e no preparo de \"remédios\" caseiros para diversos tipos de doenças e sintomas. Industrialmente, é empregado em formulações farmacêuticas, cosméticos e alimentos como corante de origem natural. Contudo, o uso da bixina ainda é limitado pela sua baixa solubilidade em água. Foi observado que a bixina pode ser dispersa em água, por meio de técnica já patenteada pelo nosso grupo, sem uso de suportes e adjuvantes. O presente projeto teve por objetivo obter a bixina purificada de um extrato comercial de semente de urucum, preparar e caracterizar a dispersão de bixina em água e aplicá-la como um carreador de fármacos, utilizando daunorrubicina como modelo e que é utilizada no tratamento de tumores. Bixina foi obtida com 95% de pureza, a partir do extrato de semente de urucum e dispersões a partir deste composto foram preparadas em soluções aquosas e estabilizadas em cerca de duas horas. Foi determinado que a dispersão em água é constituída por partículas esféricas, com diâmetro médio variando de 20 a 150 nm e potencial Zeta de -24,7 mV. A dispersão manteve-se estável quando submetida a concentrações de NaCl de até 50 mmol/L e resistente em pH ácido. Porém, a partir de pH=10, ocorreu hidrólise do éster metílico da bixina, convertendo-a em norbixina. As partículas de bixina foram capazes de incorporar o fármaco daunorrubicina, em proporção molar máxima de bixina/daunorrubicina de 2:1. Nas concentrações testadas, a incorporação da daunorrubicina à dispersão de bixina causou aumento na atividade antiproliferativa, sendo até 60% mais ativa do que a daunorrubicina livre, na concentração de 1 µg/mL. A dispersão de bixina apresentou tolerância a variações em pH e concentração salina e capacidade de incorporar e aumentar a atividade do fármaco daunorrubicina. Sendo assim, constitui-se em um potencial sistema carreador de fármaco derivado de um produto natural. / Bixin is a carotenoid found on surface of Bixa orellana L. seeds, known as annatto. Annatto seeds extracts are popularly used as a condiment for foods and remedy for several diseases and symptoms. It is also applied in pharmaceutical products, cosmetics and food industry as a natural colorant. Nevertheless, the use of bixin is still limited by its poor solubility in water. Our research group has developed and patented a technique to disperse bixin in water, with no use of stabilizers or scaffolds. In this project, we aimed to obtain purified bixin from commercial annatto seeds extracts, prepare and characterize a dispersion of bixin in water and to employ it as a drug delivery system. Daunorubicin was chosen as a drug model to be delivered by bixin dispersion. Bixin was isolated from extracts in 95% purity. Aqueous dispersions of bixin demonstrated to be stable two hours after preparation. Bixin formed spherical particles with mean diameter ranging from 20 to 150 nm and Zeta potential of -24,7 mV. The dispersion was stable in NaCl solution up to 50 mMol/L and resistant to acidic medium. However, above pH=10, hydrolysis of ester termination begins to occur, converting bixin into to norbixin and since norbixin is water-soluble, the particles dissolved in water. Bixin dispersion was able to incorporate daunorubicin, in a bixin:daunorubicin molar proportion of 2:1. In all tested concentrations, daunorubicin delivered by bixin showed higher antiproliferative activity compared to free drug, reaching 60% more acitivity at 1 µg/mL In conclusion, bixin dispersion showed good stability in large range of pH and salt concentrations, ability to incorporate daunorubicin and enhanced the antitumoral activity. Thus, it can be considered a potential drug delivery system derived from a natural product.

Wang, Jia. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2013. / Includes bibliographical references (leaves 99-115). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

In this work we present the first characterization of a Cu2+-transporting ATPase. The thermophilic bacteria Archaeoglobus fulgidus contains two genes, CopA and CopB, encoding for CPx-ATPases. CopB belongs to the subgroup IB-4 of the CPX-ATPases. These enzymes are characterized by a CPH motif in the 6th transmembrane domain and a His-rich N-terminus metal binding domain (MBD). CopB was heterologously expressed in E. coli. Membranes were prepared and used to measure activity. CopB was active at high temperature (75º C), high ionic strength and pH 5.7. The enzyme was activated by Cu2+, and in to a lesser extent by Ag+ and Cu+. CopB showed a Vmax = 5 µmol/mg/h and a high apparent affinity (K1/2 = 0.28 ± 0.09 μM) for Cu2+. Uptake of 64Cu2+ into everted vesicles was also measured in order to show that Cu2+ is not only activating the enzyme but being transported. Compared with CopB-WT, CopB-T (lacking the N-terminus MBD) did not show any difference in its activation by the different metal ions, demonstrating that the cytoplasmic MBD has no role in the metal selectivity. CopB-T also showed a 40 % decrease in the ATPase activity. CopB-WT and CopB-T presented similar levels of phosphorylation. However, CopB-T exhibited a reduced rate of dephosphorylation (slower transition from the E2P to the E2 conformation). These observations suggest a regulatory role for the cytoplasmic MBD.

FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / O Streptococcus (S.) pneumoniae à considerado como o principal agente causador de morbidade e mortalidade em crianÃas menores de cinco anos de idade. Todas as doenÃas pneumocÃcicas comeÃam com o estabelecimento da colonizaÃÃo do S. pneumoniae na nasofaringe, podendo progredir para doenÃa invasiva se as barreiras naturais forem cruzadas. Nas Ãltimas dÃcadas, o aumento do nÃmero de cepas de S. pneumoniae resistentes à antibiÃticos β-lactÃmicos e a outras classes de antimicrobianos tem dificultado o tratamento da infecÃÃo pneumocÃcica. Atualmente cerca de 13 sorotipos de S. pneumoniae respondem por mais de 85% dos isolados invasivos. A vacina pneumocÃcica polissacarÃdica conjugada 7-valente tem sido amplamente recomendada para crianÃas menores de cinco anos. Os objetivos desse estudo foram determinar a prevalÃncia de S. pneumoniae em crianÃas portadoras, a frequÃncia de isolados de S. pneumoniae de indivÃduos com infecÃÃo sistÃmica, o perfil de sensibilidade a antimicrobianos e os sorotipos mais comuns, em Fortaleza, Brasil. Os isolados de portadores foram recuperados a partir de swabs de nasofaringe de crianÃas usuÃrias de creches, enquanto que os isolados de infecÃÃo sistÃmica foram cedidos pelo LACEN-CE. Foram realizadas as ConcentraÃÃes InibitÃrias MÃnimas (CIM) para penicilina e ceftriaxona para todos os isolados, e levofloxacina apenas para os isolados de nasofaringe. Os pontos de corte das CIM foram determinados de acordo com o CLSI (2007). As sorotipagens dos isolados sistÃmicos foram realizadas pela reaÃÃo de Quellung, enquanto que a genotipagem capsular dos isolados de portadores foi realizada pela tÃcnica de multiplex PCR. De 215 crianÃas usuÃrias de creches, foram isolados S. pneumoniae em 152 (71%). As CIM de 137 isolados de portadores mostraram uma taxa resistÃncia de 71% para penicilina e de 21% para ceftriaxona. NÃo houve resistÃncia nos testes com levofloxacina. Comparado a um estudo similar, realizado hà 10 anos, em Fortaleza, nossos resultados apresentaram um aumento significativo nas taxas de resistÃncia à penicilina e ceftriaxona. De 26 isolados de nasofaringe que apresentaram resistÃncia plena, apenas, seis isolados (23%) tiveram a genotipagem capsular identificada por multiplex PCR. A incidÃncia de isolados invasivos neste estudo por ano, foi de, aproximadamente, 1 caso/100.000 hab. Dos 52 isolados, 42% apresentaram resistÃncia à penicilina e 13,5% à ceftriaxona. Os sorotipos mais comuns dos isolados sistÃmicos foram 19F (12%), 14, 3, 6A (8% cada), 4, 18C e 9V (6% cada), com cobertura estimada, tanto para vacina pneumocÃcica conjugada 7-valente quanto para a 10-valente, de 31,8%. / Streptococcus (S.) pneumoniae is considered the principal causative agent of morbidity and mortality in children younger than five years of age. All pneumococcal diseases are initiated by establishing a S. pneumoniae colonization in nasopharynx, the disease progressing to systemic disease if natural barrier are crossed. During the last decades, the increasing amount of resistant S. pneumoniae strains to beta-lactams and other classes of antimicrobials has modified the treatment of pneumococcal infection. At present, nearly 13 serotypes respond for more than 85% of invasive isolates. The 7-valent polysaccharide-conjugated pneumococcal vaccine has been widely recommended for use in children younger than five years. The aims of this study were to determine the S. pneumoniae carrier in children, the frequence of serotypes from systemic infection patients, the susceptibility profile to antimicrobials in Fortaleza, Brazil. Carrier state isolates were recovered from nasopharyngeal swabs from children attending day-care center facilities, while the isolates from systemic infection fournished by LACEN-CE. Minimal Inhibitory Concentrations (MIC) to penicillin and ceftriaxone were assessed for all isolates, and levofloxacin MIC only from nasopharyngeal isolates. MIC cut-offs were determined according to CLSI standards (2007). Serotyping of systemic isolates was performed by Quellung reaction, while capsular genotyping of carrier isolates was performed by multiplex PCR assay. OF 215 children attending day-care centers, 152 S. pneumoniae isolates were identified (71%). Penicillin MIC showed 71% of resistance, and for ceftriaxone, 21% of resistance. No resistance was found for levofloxacin MIC testing. When compared to a 10-year old similar study in Fortaleza, our results have shown a significant increase of penicillin and ceftriaxone resistance rates. Of 26 isolates tested, only six nasopharyngeal isolates (23%) were positively genotyped by multiplex PCR. The incidence of invasive isolates was 1/100,000 inhab. per year. Of 52 systemic isolates serotyped, 42% were penicillin-resistant, and 13.5% were ceftriaxone-resistant. Systemic serotypes identified were 19F, 3, 6A, 4, 18C and 9V, with a estimated coverage by the 7-valent and 10-v pneumococcal polysaccharide conjugated vaccines of 31.8%.

Crack formation under tensile forces is a major weakness of concrete. Cracks make concrete vulnerable to the extreme environment due to the ingress of water and harmful compounds from the surrounding environment. Conventional methods of crack repairing are expensive and time consuming. It is estimated that in Europe, cost related to repair works is half of the annual construction budget and the US has average annual maintenance cost for existing bridges through the year is estimated to $5.2 billion. To overcome this problem, a self-healing concrete is produced based on the application of mineral producing alkaliphilic Bacillus Subtilis (strain 168) bacteria. Metabolic activities of these bacteria on calcium-based nutrients results in precipitation of calcium carbonate, which helps to repair concrete cracks. In bacteria based self-healing concrete, the bacteria are protected in the dense cementitious matrix by encapsulating them in “bacteria-carriers”. However, the presently available bacteria-carriers are not always suitable for concrete because of their complex manufacturing procedures or high cost. With the aim to develop a more suitable bacteria-carrier, in this study feasibility of cellulose fiber as a novel bacteria-carrier for self-healing mortar is investigated. Cellulose fibers compared to other bacteria-carriers can serve the dual purpose of arresting cracks and at the same time be a bacteria-carrier in large scale concrete construction. Two types of bacterial mortar by using cellulose fiber as a carrier was prepared. For one type, nutrients were added inside the mortar mix, while for the other, nutrients were added into the curing water. The two types of composites; control and cellulose fiber reinforced concrete (CeFRC) have also been investigated for autogenous healing of concrete. The crack healing efficiency of bacterial mortars was investigated using image analysis and ultrasonic pulse velocity (UPV) test and compared with unreinforced and control cellulose fiber mortars. Variation in compressive strength for all mixes compared to control mortar is also presented in this thesis. Research shows that self-sealing mortar using cellulose fiber as a bacteria-carrier result in maximum self-healing as compared to other mixes. This study also aims to evaluate the self-healing potential and water permeability of CeFRC. Compressive strength and flexural tests were also performed to evaluate the mechanical properties of the composites. Water permeability test was used to evaluate the coefficient of permeability and the self-healing performance was investigated by using UPV and a patented self-healing test. The results indicate that the water permeability coefficient decreased by 42% (+15% or -21%) whereas the healing ratio increased at a higher rate for the initial days of healing when cellulose fibers were added in the concrete. CeFRC also results in a 7.8% increase in flexural strength. / Graduate / 2020-09-13