Submitted by Cássia Santos (cassia.bcufg@gmail.com) on 2017-07-25T13:32:13Z No. of bitstreams: 2 Dissertação - Junaine Vasques da Silva - 2002.pdf: 3727327 bytes, checksum: 4cb8c1db4d3fb95798779f39aae78673 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-07-26T12:05:00Z (GMT) No. of bitstreams: 2 Dissertação - Junaine Vasques da Silva - 2002.pdf: 3727327 bytes, checksum: 4cb8c1db4d3fb95798779f39aae78673 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-07-26T12:05:00Z (GMT). No. of bitstreams: 2 Dissertação - Junaine Vasques da Silva - 2002.pdf: 3727327 bytes, checksum: 4cb8c1db4d3fb95798779f39aae78673 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2002-12-19 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The main function of the most superficial layer of the epidermis, the Stratum Corneum (SC), is to provide a physical barrier that controls the transepidermal water loss as well as the permeation of another substances in both directions across the skin. The SC is formed by anabolically dead cells, the terminally differentiated corneocyte, and its function is essentially accomplished by forming a highly insoluble protein structure on the surface of the corneocytes, termed the cornified cell envelope, and by impeding water diffusion across the SC by mortaring the corneocytes together by layers of skin-specific lipids, essentially ceramide, cholesterol and fatty acid. In this work the cell envelope of the SC was spin labeled with a sulfhydryl-specific nitroxide reagent to investigate the water content effects upon the protein dynamics directly in the intact tissue. A two-state model for the nitroxide side chain described the coexistence of two spectral components in the electron paramagnetic resonance (EPR) spectra. The so-called strongly immobilized component, S, is associated with the EPR signal of a motionally restricted nitroxide fraction having its N-O group hydrogen bonded to protein (rigid structure) while the weakly immobilized component, W, corresponds to the signal provided by the spin labels with higher mobility (~10 times greater) exposed to the aqueous environment. The relative populations between these two mobility states, S and W, are in thermodynamic equilibrium. The standard Gibbs free energy, enthalpy and entropy changes for transferring the nitroxide side chain from the state contacting the solvent, W, to the one contacting protein, S, indicated that the reduction of the SC water content to below ~h 0.69, g H2O per g dry SC, stabilizes the protein interacting state, S. Upon decreasing the SC hydration level below ~h 0.69 the segmental motion of the polypeptide chains and the rotational motion of the spin-labeled side chain were also constrained. To test our methodology in a pure and very well known protein, we also studied the effects of two types of detergents on the bovine serum albumin (BSA). Both detergents, the anionic sodium dodecyl sulfate (SDS) and the zwitterionic N-hexadecyl-N,N-dimethyl-3-ammonium-1-propanesulfonate (HPS) increase the mobility of the protein backbone and of the nitroxide side chain. The thermodynamic parameters indicated that these detergents destabilize the protein favoring less compact conformations. This work can also be useful to improve the spectral analysis of site-directed spin labeling, especially for a more quantitative description in terms of thermodynamic parameters. / A camada mais superficial da epiderme, o Estrato Córneo (EC), tem como função principal a formação de uma barreira física que controla a perda de água do corpo bem como a permeação de outras substâncias em ambas as direções da pele. O EC é formado por células anabolicamente mortas, os corneócitos, os quais sofreram diferenciação celular terminal, e sua função é realizada formando uma estrutura de proteínas altamente insolúveis na superfície do corneócito, chamada de envelope celular, e também uma matriz lipídica, essencialmente ceramídios, colesterol e ácidos graxos, que dificultam a difusão da água. Neste trabalho, o EC foi marcado com marcadores de spin específicos para reagir com os grupos sulfidrilas das proteínas, para investigar os efeitos do conteúdo de água na dinâmica de proteínas diretamente no tecido intacto. Um modelo de dois estados para a cadeia lateral do nitróxido descreveu a coexistência de duas componentes espectrais de ressonância paramagnética eletrônica (RPE). A componente denominada fortemente imobilizada (S), surge de uma fração de marcadores com o átomo de oxigênio do nitróxido ligado à proteína (estrutura rígida) enquanto a componente fracamente imobilizada é gerada pelos marcadores com mobilidade mais alta (~10 vezes maior) e expostos ao ambiente aquoso. As populações relativas entre estes dois estados de mobilidade, S e W, estão em equilíbrio termodinâmico. Os parâmetros da termodinâmica: energia livre padrão de Gibbs, entalpia e entropia, envolvidos na transferência da cadeia lateral do nitróxido do estado W, contatando ao solvente, para o estado S, contatando a proteína, indicaram que a redução do conteúdo de água para abaixo de ~0.69g de H2O por g de EC seco, estabiliza o estado S (cadeia lateral do nitróxido dobrada sobre a cadeia principal da proteína). Ao diminuir o nível de hidratação para abaixo de ~ h 0.69 (g H2o/g EC seco) o movimento local da cadeia polipeptídica e o movimento rotacional da cadeia lateral do marcador de spin foram ambos reduzidos. Para testar nossa metodologia em uma proteína pura e bem conhecida, estudamos os efeitos de dois tipos de detergentes sobre a albumina do soro bovino (BSA). Ambos os detergentes, o aniônico dodecil sulfato de sódio (SDS) e o ziteriônico N-hexadecil-N,N-dimetil-3-amônio-1-propanosulfonato (HPS) aumentaram a mobilidade da cadeia principal da proteína e da cadeia lateral do nitróxido. Os parâmetros termodinâmicos indicaram que estes detergentes desestabilizam a proteína favorecendo conformações menos compactas. Os resultados do presente trabalho também podem contribuir para aprimorar a

Submitted by Erika Demachki (erikademachki@gmail.com) on 2014-10-13T21:33:03Z No. of bitstreams: 2 Dissertação - Adriana Pereira Mundim Guedes - 2013.pdf: 2999561 bytes, checksum: 755cb864a8446e6ff5c334be00ea5367 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Jaqueline Silva (jtas29@gmail.com) on 2014-10-16T18:47:44Z (GMT) No. of bitstreams: 2 Dissertação - Adriana Pereira Mundim Guedes - 2013.pdf: 2999561 bytes, checksum: 755cb864a8446e6ff5c334be00ea5367 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2014-10-16T18:47:44Z (GMT). No. of bitstreams: 2 Dissertação - Adriana Pereira Mundim Guedes - 2013.pdf: 2999561 bytes, checksum: 755cb864a8446e6ff5c334be00ea5367 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2013-09-13 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Motivated by the perspective of ruthenium complexes to be used in cancer treatment, our research group has tested the hipotesis that some complexes of Ru (III) are able to interact with serum proteins, particularly albumin and transferrin. The Complex cis- [RuCl2(NH3)4]Cl (CTRu(III)) have been tested against different kind of tumor cells, obtaining good results. Starting from promising results obtained with this compound, subsequent studies are required to understanding the mechanism by which it exerts specificity for tumor cells. In this article, we report the first application of absorption UV-Vis, Fluorescence and Electron Paramagnetic Resonance (EPR) spectroscopy, to study the complex CTRu(III) interaction with human serum albumin (hsA) and bovine serum albumin (bsA). Fluorescence measurements revealed strong proteinsbound complex with Ksv of 1.32 x 105 and 3.71 x 105 for hsA and bsA, respectively. EPR spectra from mono-nuclear Ru(III) complexes in buffer, showed a significant decrease in the overall signal intensity following the first aquation step, is consistent with the formation of oxo-bridged Ru(III) dimers. EPR spectra revealed that the BSA very rapid binding to the protein via covalent binding through ligand-exchange with protein side chains, likely with histidine imidazoles. On the other hand, the complex binds non-covalently in hsA, probably as a product of the oligomerization of the complex in hemin-biding pocket. Furthermore, two species are slowly formed by covalent binding of the complex with the histidine residues, producing a species of axial symmetry and the other rhombic symmetry. These bonds seem to arise from the interaction of the complex with the histidine residue located in the binding Sudlow’s site II. / Motivado pela perspectiva de complexos de rutênio podem ser utilizados no tratamento do câncer, o nosso grupo de pesquisa testou a Hipótese que alguns complexos de Ru (III) são capazes de interagir com as proteínas do soro, particularmente albumina e transferrina. O complexo de cis-[RuCl2(NH3)4]Cl (CTRu(III)) foi testado contra diferentes tipos de células tumorais, obtendo bons resultados. A partir de resultados promissores obtidos com este composto, estudos subsequentes são necessários para a compreensão do mecanismo pelo qual ele exerce sua especificidade para células de tumor. Neste artigo, apresentamos a aplicação de espectroscopia de absorção UV-vis, fluorescência e ressonância paramagnética eletrônica (RPE), para estudar a interação do complexo CTRu(III) com albumina sérica humano (hsA) e a albumina sérica bovina (bsA). Medidas de fluorescência revelaram uma forte ligação do complexo com as proteínas com Ksv de 1,32 x 105 e 3,71 x 105 para hsA e bsA, respectivamente. Espectros de RPE de complexos de Ru (III) mono-nucleares em tampão mostraram um decréscimo significativo na intensidade do sinal global após a primeira passo de aquação, que é consistente com a formação de dímeros de oxo complexos de Ru (III). Os espectros de RPE revelaram que a ligação à bsA é muito rápida, a ligação covalente à proteína ocorre através de troca dos ligantes com cadeias laterais de proteínas, provavelmente com o imidazol da histidina. Por outro lado, o complexo se liga não covalentemente na hsA, provalente como produto da oligomerização do complexo no bolso de ligação hemin. Além disso, duas espécies são formadas lentamente por ligação covalente do complexo com os resíduos histidina, produzindo uma espécie de simetria axial e a outra de simetria rômbica. Essas ligações parecem surgir pela interação do complexo com o resíduo histidina localizado no sítio de ligação Sudlow II.

No description available.

Welding of duplex stainless steel pipeline material for the oil and gas industry is now common practice. To date, research has been conducted primarily on the parent material and heat affected zones in terms of its susceptibility to various forms of corrosion. However, there has been little research conducted on the degree of sensitisation of the various successive weld layers, namely the root, fill and cap layers. The focus of this research study was to: (i) provide an in-depth microstructural analysis of the various weld passes, (ii) study the mechanical properties of the weld regions; (iii) determine degree of sensitisation of the various weld passes; and (iv) investigate the residual stress levels within the various regions/ phases of the welds. Four test conditions were prepared using manual Gas Tungsten Arc Welding with 'V' and 'U' bevel configuration. Structural analysis consisted of (i) optical microscopy, scanning electron microscopy and magnetic force microscopy; (ii) ferrite determination using Magna-Gauge, Fischer Ferrite-scope and Point Count method. Mechanical testing consisted of Vickers hardness measurements, Charpy impact studies and transverse tensile testing. The degree of sensitisation was determined by three test methods: a modified ASTM A262, ASTM A923 and a modified Double Loop Electrochemical Potentiodynamic Reactivation (DL-EPR) test. Residual stress levels were determined using two neutron diffraction techniques: a reactor source and a time of flight spallation source. Microstructure observed by optical microscopy and magnetic force microscopy shows the formation of both fine and coarse structures within the weld metal. There was no evidence of secondary austenite, being present in any of the weld metal conditions examined. In addition, no detrimental intermetallic phases or carbides were present. The DL-EPR test results revealed that the fill layer regions for all four conditions and the base material showed the highest values for Ir/Ia and Qr/Qa. All four test conditions passed the ASTM A262 and A923 qualitative type tests, even under restricted and modified conditions. Residual stress measurements by neutron diffraction conducted at Lucas Heights Hi-Flux Reactor revealed that the ferrite phase stress was tensile in the heat affected zones and weld, and appeared to be balanced by a local compressive austenite phase stresses in the normal and transverse directions. Residual stress measurements by neutron diffraction conducted at Los Alamos Nuclear Science Centre revealed that in the hoop direction, ferrite (211) and austenite (311) exhibit tensile strains in the weld. In the axial and radial direction, the strains for both phases were more compressive. Correlations between the degree of sensitization and microstructural changes / ferrite content were observed. Higher degrees of sensitization (Ir/Ia and Qr/Qa) were associated with reduced ferrite (increased austenite) content. Correlations between the stresses generated, the evolved microstructures and degree of sensitization were evident. Stresses within the cap region were generally shown to be of a tensile nature in the transverse and longitudinal direction. In summary, the study has shown that correlations exist between the weld microstructure, susceptibility to sensitisation and levels / distribution of internal stresses within the weld regions.

La fonction d'onde de deux particules après<br />une interaction mutuelle ne peut pas, en général, être exprimée<br />comme le produit d'états individuels des particules. Pour qu'une<br />collision atomique puisse produire une intrication<br />maximale, il faut cependant des conditions exceptionnelles. Ces<br />conditions sont réunies dans notre dispositif, où l'interaction<br />dipôle-dipôle entre atomes de Rydberg "à deux niveaux" peut être<br />stimulée par une cavité supraconductrice non-résonnante. En<br />exploitant cet effet, nous avons réalisé des états fortement<br />intriqués dans des collisions binaires avec des paramètres<br />d'impact de l'ordre du millimètre. L'angle de collision peut être<br />varié en modifiant le désaccord atomes-cavité, ce qui nous a<br />permis d'observer l'échange d'énergie réversible et cohérent<br />(oscillation de Rabi) entre deux atomes. Par un choix opportun des<br />valeurs des paramètres, nous avons en particulier réalisé et testé<br />une paire 'EPR' d'atomes. La relative insensibilité de cette<br />méthode d'intrication au facteur de qualité du résonateur rend sa<br />fidélité compatible avec des expériences de violation des<br />inégalités de Bell ainsi qu'avec la manipulation cohérente d'un<br />nombre plus important d'atomes. Nous présentons en outre une<br />nouvelle procédure de fabrication des cavités micro-ondes. Les<br />nouvelles cavités devraient en particulier permettre l'extension<br />des études sur l'intrication aux champs confinés dans deux<br />résonateurs séparés.

A liquid Cooled below its normal freezing temperature is known as a supercooled liquid. On further cooling, supercooled liquids crystallize to thermodynamically stable, ordered structures. Alternatively, if the cooling rate is fast enough, the crystallization may be avoided altogether. Below a particular temperature during rapid cooling the liquid will solidify into a disordered, amorphous phase -also known as the glassy phase of matter. This particular temperature is termed the ”glass transition temperature” (Tg). Unlike a crystalline solid, a glass is neither a thermodynamically stable phase nor does it possess long range molecular ordering. Very slow structural relaxation (in the time scale of ∼ 100 s) is always present in the glassy phase. Thus, this phase is often referred to as a metastable phase of matter. Experimental and theoretical studies related to the behavior of supercooled liquids are the subject matter of many investigations for the last few decades [1]. These studies find their applications in diverse fields such as geology, cryopreservation, glaciology and atmospheric science. However, properties of supercooled liquids and the corresponding amorphous phase are not completely understood at present, particularly for hydrogen bonded (H-bonded) systems. This thesis concerns both the crystallization and the glass formation process of H-bonded systems. The systems of interest are water, the commonly accepted universal solvent, and the aqueous binary mixture of glycerol and water. The technique of molecular probing is often used to study the cooperativety and rotational diffusion of supercooled liquids and for determination of the glass transition temperature. For the present set of work, a molecular probe technique called spin probe ESR is extensively used. Electron paramagnetic resonance or electron spin resonance (EPR/ESR) measures the electronic energy level separation and is well known for the high sensitivity. All of the systems studied in the present set of work are diamagnetic. This issue is circumvented by dissolving paramagnetic spin probe molecules, which are usually organic free radicals with one N-O group, into the systems. Spin probes are added in very low concentrations (~10-3M) to minimize the effect on the host system and also to avoid mutual interactions between them. The unpaired electron delocalized in the direction of the N-O bond serves as the paramagnetic center required for an ESR experiment. The splitting of electron energy level due to the external magnetic field (Zeeman splitting) can give rise to resonance absorption of energy if exposed to a microwave of appropriate frequency. There is also a magnetic coupling (hyperfine) between the spin of the unpaired electron and nuclear spin of the nearby nitrogen atom. The hyperfine coupling splits each electron energy levels, to the first order, symmetrically into three levels. The transitions between these levels -subject to appropriate selection rules -give rise to the ESR spectrum [2]. The spectral shape in a magnetic field sweep ESR experiment appears complex if randomly oriented spin probes are dispersed in an amorphous or polycrystalline solid matrix. The high degree of mobility in probe molecules, present in a liquid solution, can average out the individual anisotropy of magnetic tensors to get a spectrum of three equally spaced liens. Experiments can be performed spanning a spin probe reorientation timescale of 10-7-10-12 s typically in the temperature range of 4.2 -300K. In chapter one we have given a brief overview of the supercooled liquids and the phase transitions related to the present work. Particular emphasis has been given to the dynamical features of the supercooled liquid close to its glass transition temperature and their classification based on the degree of ’fragility’ [3]. Brief general introductions of the systems studied in each of the following chapters are also provided. Then, the details of ESR spectroscopy and a quantum mechanical picture of the method of spin probe ESR have been discussed [4]. A separate section has been devoted to the numerical and analytical methods used to analyze the spectrum to extract information related to the spin probe dynamics [5]. The chapter concludes with a description of the ESR spectrometer. In chapter two we have studied the glass transition and dynamics of the supercooled water by the method of spin probe ESR. The vitrification has been done by direct exposure of the bulk water sample, doped with the spin probe TEMPOL, to the liquid helium flow. The vitrified matrix turns into the ultraviscous liquid above the putative glass transition temperature of ~136 K which further transforms to cubic ice (Ic) above TX ~150 K. The supercooled fraction of water, along with the spin probes which are treated as impurities by the crystallized surroundings, remain trapped inside the veins or triple junctions of the ice grains which serve as the interfacial reservoir of impurities in a polycrystalline ice matrix. The spectra for the entire temperature range have been analyzed with the help of in-depth computation by modelling the reorientation of TEMPOL in terms of the jump angle θs and the rotational correlation time τ [5]. This model, based on a homogeneous mobility scenario of the spin probe, works nicely except in the temperature range of 140-180 K. Dynamical heterogeneity (DH) is apparent in this temperature range and a more mobile (fast) component, as compared to the one corresponding to the very slow dynamics of TEMPOL at lower temperatures (slow), is observed. The relative weight of the fast and the slow component changes with temperature and above ~180 K the entire spectrum changes into the motionally narrowed triplet. The temperature dependence of the slow component of τ shows a change in slope at a temperature close to the putative glass transition temperature of water. The fast component of τ exhibits a fragile, i.e. non-Arrhenius character at high temperature with a crossover to a strong, i.e. Arrhenius behavior below ~225 K, close to the hypothesized fragile-to-strong crossover (FSC) for water at TFSC ~228 K. The breakdown of the Debye-Stokes-Einstein (DSE) law is observed when the τ values are combined with the available viscosity data of water to evaluate the DSE ratio, paralleling the SE breakdown which has recently been observed in nanoconfined water [6]. The dynamical heterogeneity is thought to be closely associated with the static structural heterogeneities of supercooled water. The existence of large scale structural fluctuations spanning a range of low-and high-density phases of liquid water have been associated with the heterogeneous dynamics sensed by TEMPOL. Motivated by the Arrhenius like behavior of the slow component, it has been identified with the low density liquid (LDL). The fragile nature of the fast component at high temperature may be identified with that of the high density liquid (HDL) which is the predominant fraction in liquid or weakly supercooled water [6]. Chapter three reports the studies on freezing and dynamics of the supercooled water trapped inside the veins of a polycrystalline ice matrix by dissolving spin probes TEMPO and TEMPOL into it. When a millimolar spin probe aqueous solution is cooled below the freezing point of water, the spin probes -driven by the mechanism described above migrate to the liquid environment inside the ice veins. Local concentration of the probe molecules inside the veins can go up to 1-10 M [7]. Bulk crystallization is evident in differential scanning calorimetry (DSC) studies whereas the liquid environment of the spin probe below the bulk freezing is confirmed by its narrow triplet ESR spectrum. A sudden collapse of this narrow triplet into a single broad line indicates the freezing of the trapped water fraction which usually happens well below the DSC freezing point for both the probes. The spin probe detected freezing point of this interstitial water is found to be largely dependent on the properties and the amount of the dissolved probe molecules. An explanation is sought in terms of the ’destructuring effect’ on the tetrahedral ordering of the water H-bond network by both the high local concentration of the spin probes and the hydrogen bond strength, formed between the water and the spin probe molecules through the polar groups of the latter [8, 9]. These two factors are thought to play important roles in determining the reorientational dynamics of the spin probe molecules, as well. The rotational correlation times of the two probes exhibit a crossover owing to the different mobility of their salvation shells in the more ordered supercooled water. The observed relaxation behavior of this confined water using the probe TEMPO, which has little effect on water H-bond network, is found in agreement with the previous experimental investigations on water confined in a nanochannel [10]. In chapter four, the glass transition, relaxation and the free volume of the glycerol-water (G-W) system are studied over the glycerol concentration range of 5 -85 mol% with TEMPO as the spin probe. G-W mixture is intrinsically inhomogeneous due to the well established phase segregation below a critical glycerol concentration of 40 mol%. In the inhomogeneous regime the water molecules tend to form cooperative domains besides the mesoscopic G-W mixture [11]. Samples are quenched by rapid cooling down to 4.2 K inside the spectrometer cryostat. Spectra were recorded on slow heating of the sample in the temperature range of 130 -305 K. The glass transition temperature is correlated to the sharp transition of the extrema separation of the ESR spectrum. The glass transition temperatures are found to follow a concentration dependence which is closely associated to the mesoscopic inhomogeneities of the G-W system. The steady enhancement in fragility of the G-W system with the addition of water is evident from the temperature dependence of the spin probe correlation time τ for the entire concentration range. In the temperature range of 283 -303 K, the DSE law is followed i.e. the spin probe reorientation process is found to be strongly coupled to the system viscosity. In this regime, the τ values have been used along with the available viscosity data to calculate the effective volume V of the spin probe for the entire concentration range. The spin probe effective volume is a measure of the available free volume of the host matrix. A drastic change in the quantity is seen in the vicinity of the 40 mol% glycerol concentration owing to a similar structural change of the matrix due to the formation of mesoscopic scale inhomogeneities below the critical concentration [12]. The thesis concludes with a discussion about the possible future directions of research.

Tyrosine Hydroxylase (TyrH) is a pterin-dependent mononuclear non-heme iron oxygenase. TyrH catalyzes the hydroxylation reaction of tyrosine to dihydroxyphenylalanine (DOPA). This reaction is the first and the rate-limiting step in the biosynthesis of the catecholamine neurotransmitters. The active site iron in TyrH is coordinated by the common facial triad motif, 2-His-1-Glu. A combination of kinetic and spectroscopic techniques was applied in order to obtain insight into the catalytic mechanism of this physiologically important enzyme. Analysis of the TyrH reaction by rapid freeze-quench Mossbauer spectroscopy allowed the first direct characterization of an Fe(IV) intermediate in a mononuclear nonheme enzyme catalyzing aromatic hydroxylation. Further rapid kinetic studies established the kinetic competency of this intermediate to be the long-postulated hydroxylating species, Fe(IV)O. Spectroscopic investigations of wild-type (WT) and mutant TyrH complexes using magnetic circular dichroism (MCD) and X-ray absorption spectroscopy (XAS) showed that the active site iron is 6-coordinate in the resting form of the enzyme and that binding of either tyrosine or 6MPH4 alone does not change the coordination. However, when both tyrosine and 6MPH4 are bound, the active site becomes 5-coordinate, creating an open site for reaction with O2. Investigation of the kinetics of oxygen reactivity of TyrH complexes in the absence and presence of tyrosine and/or 6MPH4 indicated that there is a significant enhancement in reactivity in the 5-coordinate complex in comparison to the 6-coordinate form. Similar investigations with E332A TyrH showed that Glu332 residue plays a role in directing the protonation of the bridged complex that forms prior to the formation of Fe(IV)O. Rapid chemical quench analyses of DOPA formation showed a burst of product formation, suggesting a slow product release step. Steady-state viscosity experiments established a diffusional step as being significantly rate-limiting. Further studies with stopped-flow spectroscopy indicated that the rate of TyrH reaction is determined by a combination of a number of physical and chemical steps. Investigation of the NO complexes of TyrH by means of optical absorption, electron paramagnetic resonance (EPR) and electron spin echo envelope modulation (ESEEM) techniques revealed the relative positions of the substrate and cofactor with respect to NO, an O2 mimic, and provided further insight into how the active site is tuned for catalytic reactivity upon substrate and cofactor binding.

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Gold nanoparticles exhibit a combination of physical, chemical, optical, and electronic properties unique from all other nanotechnologies. These structures can provide a highly multifunctional platform with which to diagnose and treat diseases and can dramatically enhance a variety of photonic and electronic processes and devices. The work herein highlights some newly emerging applications of these phenomena as they relate to the targeted diagnosis and treatment of cancer, improved charge carrier generation in photovoltaic device materials, and strategies for enhanced spectrochemical analysis and detection. Chapter 1 introduces the reader to the design, synthesis, and molecular functionalization of gold nanotechnologies, and provides a framework from which to discuss the unique photophysical properties and applications of these nanoscale materials and their physiological interactions in Chapter 2. Chapter 3 discusses ongoing preclinical research in our lab investigating the use of near-infrared absorbing gold nanorods as photothermal contrast agents for laser ablation therapy of solid tumors. In Chapter 4, we present recent work developing a novel strategy for the targeted treatment of hormone-dependent breast and prostate tumors using multivalent gold nanoparticles that function as highly selective and potent endocrine receptor antagonist chemotherapeutics. In Chapter 5, we discuss a newly-emerging tumor-targeting strategy for nanoscale drug carriers which relies on their selective delivery to immune cells that exhibit high accumulation and infiltration into breast and brain tumors. Using this platform, we further investigate the interactions of nanoscale drug carriers and imaging agents to a transmembrane protein considered to be the single most prevalent and single most important contributor to drug resistance and the failure of chemotherapy. Chapter 6 presents work from a series of studies exploring enhanced charge carrier generation and relaxation in a hybrid electronic system exhibiting resonant interactions between photovoltaic device materials and plasmonic gold nanoparticles. Chapter 7 concludes by presenting studies investigating the contributions from so-called “dark” plasmon modes to the spectrochemical diagnostic method known as surface enhanced Raman scattering.

Στην παρούσα Διπλωματική Εργασία μελετήθηκε το σύστημα αντίδρασης του βαναδίου, V, με τον υποκαταστάτη δι-2-πυρίδυλο κετόνη, (py)2CO. Πρόκειται για έναν εξαιρετικά δημοφιλή υποκαταστάτη τον οποίο πολλές ερευνητικές ομάδες, όπως και η δική μας, έχουν μελετήσει με πολλά μεταλλοϊόντα, εκτός του βαναδίου. Η χημεία της δι-2-πυρίδυλο κετόνης βασίζεται στο γεγονός ότι η καρβονυλική της ομάδα μπορεί να υποστεί προσβολή από διάφορα πυρηνόφιλα (π.χ.H2O, ROH) και να σχηματίσει την ημικεταλική ή/και τη 1, 1-διολική μορφές της. Η μελέτη αυτού του συστήματος οδήγησε στη σύνθεση και το χαρακτηρισμό συμπλόκων του βαναδίου σε διάφορες οξειδωτικές βαθμίδες. Χαρακτηριστικό παράδειγμα αποτελεί το μικτού σθένους εννεαπυρηνικό σύμπλοκο (Et3NH)2[VIIVV8O18{(py)2CO2}4]∙2MeCN, με τον υποκαταστάτη να βρίσκεται στην διπλά αποπρωτονιωμένη 1, 1-διολική μορφή του. Ακόμη από το γενικό σύστημα αντίδρασης απομονώθηκαν σύμπλοκα VIV2, VV2, VIV3, VV4, VV12 και VΙV2 VV8 με τον υποκαταστάτη να βρίσκεται σε διάφορες μορφές. Μελετήθηκαν οι μαγνητικές ιδιότητες των συμπλόκων του τετρασθενούς βαναδίου. Η συμμετοχή ωστόσο του μετάλλου και των ενώσεων του σε διάφορες βιολογικές διεργασίες, όπως στη ρύθμιση του μεταβολισμού, στα ενεργά κεντρά μεταλλοενζύμων καθώς και η ικανότητά του να ενισχύει τη δράση της ινσουλίνης, μας έδωσαν το ερέθισμα να μελετήσουμε συγκεκριμένα την αντικαρκινική συμπεριφορά επιλεγμένων συμπλόκων που παρασκευάσαμε σε διάφορες σειρές καρκινικών κυττάρων. / In the present Diploma Work we have studied reactions of vanadium, V, sources and di-2-pyridyl ketone, (py)2CO. This ligand is very popular and its reactions with several metal ions have been investigated by many groups, including our group. No V complexes of (py)2CO have been reported. The chemistry of di-2-pyridyl ketone is based on the fact that its carbonyl group can be attacked by various nucleophiles (e.g. π.χ.H2O, ROH) to give the hemiketal or/and 1,1-diol forms. The systematic investigation of the V/(py)2CO reaction system has led to the synthesis and characterization of V compounds with the metal in various oxidation states. A characteristic example is the mixed-valence, enneanuclear complex (Et3NH)2[VIIVV8O18{(py)2CO2}4]∙2MeCN, in which the ligand participates in the doubly deprotonated 1,1-diolate form. From this general reaction system, VIV2, VV2, VIV3, VV4, VV12 και VΙV2 VV8 species have also been isolated. The magnetic properties of the VΙV complexes have also been studied. The participation of V compounds in important biological processes gave us the stimulus to study the antiproliferative activity of selected complexes in various cancer cell lines.