Student Number : 0002483R - MSc dissertation - School of Molecular and Cell Biology - Faculty of Science / Isothermal titration calorimetry was used to investigate the forces that drive ligand binding to the active site of rGST M1-1. In an attempt to gain insight into the recognition of non-substrate ligands by GSTs, this study also investigates interactions between rGST M1-1 and ANS, a non-substrate ligand. At 25 °C, complex formation between rGST M1-1 and GSH, GSO3 -, and S-hexylglutathione is characterised by a monophasic binding isotherm with Kd values of 38.5 mM, 2.1 mM and 0.2 mM, respectively. One molecule of each ligand is bound per monomer of rGST M1-1. Binding of these ligands is enthalpically favourable and entropically unfavourable with a resultant favourable Gibbs free energy, overall. The effects of temperature and buffer ionisation on the energetics of binding were studied. The enthalpic and entropic contributions for all three ligands exhibited temperature dependence over the temperature range investigated (5-30 °C). The Gibbs free energy showed negligible changes with increasing temperature due to enthalpy-entropy compensation. The temperature dependence of the binding enthalpy yielded heat capacity changes of – 2.69 kJ/mol/K and –3.68 kJ/mol/K at 25 °C for GSH and S-hexylglutathione binding and –1.86 kJ/mol/K overall for GSO3 -. The linear dependence of DH on temperature for GSO3 - binding to rGST M1-1 suggests the formation of a more constrained complex which limits the fluctuations in conformations of the mu-loop at the active site. The non-linear dependence of DH on temperature for GSH and Shexylglutathione binding to the enzyme suggests the formation of a complex that samples different bound conformations due to the mobility of the mu-loop even after ligand is bound. Calorimetric binding experiments in various buffer systems with different ionisation enthalpies suggest that the binding of GSH to rGST M1-1 is coupled to the deprotonation of the thiol of GSH while GSO3 - binding to rGST M1-1 is independent of the buffer ionisation. At 25 °C, the rGST M1-1#1;ANS association is represented by a monophasic binding isotherm with one molecule of ANS bound per monomer of rGST M1-1. The interaction is both enthalpically and entropically driven with a Kd value of 27.2 mM representing moderate affinity. The effect of temperature on the interaction was investigated over the temperature range of 5-30 °C. The linear dependence of the binding enthalpy on temperature indicates that no significant structural changes occur upon binding of ANS to the enzyme (DCp = -0.34 kJ/mol/K). The change in heat capacity associated with the interaction can be attributed to the burial of the polar sulphonate group of ANS and the exposure of the anilino and naphthyl rings to solvent as well as the possibility of weak electrostatic interactions between ANS and residues at the active site. The effect of ethacrynic acid, GSH, GSO3 - and S-hexylglutathione on the fluorescence of ANS was investigated in order to obtain some idea as to the location of the ANS binding site on rGST M1-1. ANS was displaced by GSO3 -, S-hexylglutathione and ethacrynic acid, while no displacement occurred upon binding of GSH to the active site of rGST M1-1. Displacement studies and molecular docking simulations indicate that ANS binds to the H-site of rGST M1-1 and the possibility of a second binding site for the molecule cannot be ruled out.

In striving to produce high quality cast components from recycled aluminium alloys,imperfections have to be considered, because recycled aluminium usually containsmore of it. However, there are great energy savings to be made by using recycledaluminium; as little as 5% of the energy needed for primary aluminium productionmay be required. High quality castings are dependent on, besides alloy chemistry, bothmelt quality and the casting process; the focus of this work is related to the meltquality.This thesis aims to increase knowledge about imperfections, foremost about Fe-richparticles, oxides/bifilms, and porosity. Experiments were performed at industrialfoundry facilities and in a laboratory environment. Melt quality was evaluated byproducing samples with the reduced pressure test (RPT), from which both densityindex (DI) and bifilm index (BI) could be measured, results that were related to tensiletest properties. Data from tensile test samples were analysed, and fracture surfacesand cross sections were studied in both light microscope and in scanning electronmicroscope (SEM). For the purpose of investigating nucleation of primary Fe-richparticles (sludge) differential scanning calorimetry (DSC) was used.In the analysis of results, a correlation between the morphology of particles and tensileproperties were found. And elongated Fe-rich β-particles were seen to fracturethrough cleavage towards the centre. However, DI and BI have not been possible torelate to tensile properties.The nucleation temperature of primary Fe-rich particles were found to increase withincreased Fe, Mn, and Cr contents, i.e. the sludge factor (SF), regardless of cooling rate.For a set SF, an increase of cooling rate will decrease the nucleation temperature.

An isothermal calorimeter controlled by the Peltier effect has been designed and constructed in order to investigate reaction rates in solventwater mixtures. Because a thermal method was used a constant temperature environment was essential and this was achieved by using a water bath controlled to + 0.0010C. This callorinieter has been used to study the alkaline hydrolysis of methyl acetate in dimethylsulphoxide, and tetrahydrofuran - water mixtures at 15, 25 and 35 [degrees]C. The results of other investigations on similar reactions have been reviewed and an attempt has been made to correlate the electrostatic theories of Laidler and Eyring, and Amis and jaffe with these results. Finally, because it appears that specific solvent interactions play a major part in the reaction rates the role of water in the reaction mechanism has been examined. A mechanistic explanation has been proposed in order to correlate the rate of reaction with the composition of water-solvent mixtures which justifies the Laidler and Eyring treatment of solvent effects on ion-molecule reactions.

No description available.

L'objectif de cette thèse était d'aboutir à une meilleure compréhension du comportement thermique d'un calorimètre, différentiel non adiabatique permanent, dédié à la mesure de l'échauffement nucléaire en MTR, puis à une optimisation de ce capteur et des méthodes de mesures associées et enfin à une proposition d'une configuration calorimétrique « miniaturisée ». Du fait du principe même de la calorimétrie (quantification d'énergie à partir de mesures de températures), une approche analytique ciblant les aspects thermiques a été conduite. Cette thèse a consisté à la conception, au développement et à l'exploitation de nouveaux outils analytiques thermiques expérimentaux et numériques. Un modèle thermique 2D axisymétrique résolu par méthode des éléments finis (code CAST3M) a été mis en œuvre, validé et utilisé en conditions non irradiées ou irradiées dans le cadre d'une étude paramétrique complète portant sur la réponse de différentes configurations calorimétriques. Ces travaux ont permis le dimensionnement d'un capteur plus sensible adapté aux conditions ciblées lors des premières campagnes d'irradiation en périphérie du réacteur OSIRIS (< 2W/g). Ces travaux ont également permis de définir une nouvelle cellule calorimétrique à échange directionnel radial, plus compacte pour des expériences futures en cœur du réacteur RJH à fort échauffement nucléaire (20W/g). Un dispositif expérimental a été conçu afin d'étudier le calorimètre plus sensible pour différentes contraintes thermiques (puissance injectée, température du fluide caloporteur) et fluidiques (intensité de la convection forcée) imposées. / The aim of this thesis was to reach a better understanding of the calorimeter thermal behavior of a calorimeter, differential non-adiabatic permanent, dedicated to the nuclear heating measurement in MTR, an optimization of the sensor and measurement methods associated and finally a suggestion of a calorimetric configuration "miniaturized." Because of the principle of calorimetry (quantification of energy from temperature measurements), an analytical approach targeting thermal aspects was carried out. This thesis consisted of designing, development and exploitation of new experimental and numerical analytical thermal tools. A 2D axisymmetric thermal model solved by finite element method (CAST3M code) was implemented, validated and used in conditions unirradiated or irradiated through a complete parametric study related to the response of different calorimetric configurations. This work has allowed to design a more sensitive sensor adapted to conditions targeted for the first irradiation campaigns on the OSIRIS reactor reflector (<2W/g). These studies have also allowed to define a new calorimetric cell with radial directional exchange, more compact for future experiments in the RJH core with high nuclear heating (20W/g). An experimental set up was designed to study the most sensitive calorimeter in different thermal conditions (injected power, coolant temperature) and flow conditions (intensity of forced convection) imposed.

A Dissertation Submitted to the Faculty of Medicine, University of the Witwatersrand, Johannesburg, in Partial Fulfilment of the Requirements for the Degree of Master of Pharmacy Johannesburg, March 1994 / Preformulation and formulation study of dexchlor- pheniramine maleate (DCPM) for it's inclusion into a gelforming sustained release dosage form was investigated. A modification of the USP apparatus 2 is proposed as an alternative to currently recommended USP dissolution apparatus for floating, gelforming drug delivery systems. In addition, the role of magnesium stearate and talc as dissolution retardants in controlled release matrix tablets is investigated, through application of a factorial design. / IT2018

O condutor protônico zirconato de bário dopado com ítrio (BaZr1-xYxO3-&delta;, BZYx) é um material promissor para a aplicação como eletrólito sólido em células a combustível operacional em temperaturas intermediárias (400 a 700 oC). No entanto, sua natureza refratária (ponto de fusão ~ 2600 oC) faz com que para sua densificação, necessária para sua aplicação como eletrólito, sejam necessários altas temperaturas e longos tempos de tratamento térmico (1600 a 1800 °C por 24 a 48 h). Tais condições extremas causam um desvio da estequiometria de bário que afeta a química de defeitos do material e, consequentemente a diminuição da condutividade protônica do BZYx. Portanto, o processamento desse eletrólito sólido em menores temperaturas, preservando sua estequiometria, formando uma microestrutura densa e com baixa resistividade inter-granular são os principais objetivos e desafios da comunidade científica. Visando aumentar a sinterabilidade das partículas, o BZY foi preparado pelo método dos peróxidos oxidantes (OPM). O procedimento experimental original do OPM foi modificado e otimizado para viabilizar a formação do BZYx, com x = 10 a 50 mol% de Y3+. Dentre as modificações, a síntese foi feita com e sem o controle da atmosfera, em câmara de luvas sob atmosfera de nitrogênio e ao ar, respectivamente. As propriedades estruturais, morfológicas, térmicas, termodinâmicas e elétricas das composições de BZYx foram investigadas. As amostras produzidas foram calcinadas em diversas temperaturas e investigadas quanto à sua sinterabilidade e densificação. Os pós de BZYx, com x = 10 a 50 mol% de Y3+, produzidos com controle da atmosfera foram investigados quanto às suas propriedades termodinâmicas. Os valores de entalpia de formação a partir dos óxidos (&Delta;Hf,ox) foram calculados com os dados obtidos por calorimetria de dissolução a alta temperatura. As amostras de BZY10 e BZY20 produzidas com controle da atmosfera atingiram condutividade elétrica total de 1,6 x 10-3 e 1,3 x 10-3 S/cm a 530 oC, respectivamente. A alta resistividade inter-granular contribui para a alta resistividade total das amostras. A análise por espectroscopia Raman e os valores de &Delta;Hf,ox obtidos sugerem que para valores de Y3+ > 20 mol% ocorrem interações defeito-defeito na estrutura cristalina, causando à diminuição de sítios efetivos para a hidratação e a diminuição da mobilidade dos prótons na estrutura e, consequentemente, a diminuição da condutividade protônica total. / The proton conductor oxide yttrium doped barium zirconate (BaZr1-xYxO3-&delta;, BZYx) is a promising solid electrolyte for solid oxide fuel cells (SOFC) operating at intermediate temperatures (400 to 700 oC). However, the BZY refractory nature (MP ~ 2600 oC) inhibits the achievement of the densification needed for application in SOFCs (relative density &ge; 95% T.D.), requiring long dwell times and high temperatures (T &ge; 1600 oC, t &ge; 24 h). Those extreme conditions cause barium stoichiometry deviation, which affects the defect chemistry and the depletion of proton conductivity. Therefore, BZY processing in less aggressive conditions, preserving cation stoichiometry, leading to dense microstructures with low intergranular resistivity are the great challenges of the scientific community nowadays. Aiming to increase particles sinterability, BZYx (x = 10 to 50 mol% of Y3+), solid solutions where synthesized by the Oxidant Peroxide Method (OPM). The original OPM experimental procedure was modified to allow the BZY formation with different dopant content. One of the modifications was to carry out the synthesis under laboratory and nitrogen atmospheres. The study of structural, thermal, morphological, thermochemical and electrical properties of all samples was performed. The samples where calcined at different temperatures and the particles sinterability and densification were also investigated. The thermochemical properties of BZYx solid solutions were investigated by high temperature oxide melt solution calorimetry, for evaluation of the formation enthalpies (&Delta;Hf,ox). The total electrical conductivity of the BZY10 and BZY20 sintered samples synthesized under nitrogen was 1.6 x 10-3 and 1.3 x 10-3 S/cm at 530 oC, respectively. The blocking of charge carriers at interfaces contributes to the low total electrical conductivity. Raman spectroscopy analysis and the evaluated &Delta;Hf,ox values obtained suggest that from 20 mol% Y3+, defect interaction might happen, leading to vacancy clustering. This effect might cause the depletion of mobile oxygen vacancies, affecting the mobility of protons, with a decrease in proton conductivity.

O objetivo deste estudo é realizar a caracterização térmica, resistência à flexão e o grau de conversão de um compósito nanoparticulado (Filtek Supreme XT: FT) e um microhíbrido (Esthet X: ET), cor A2, submetidos a diferentes tratamentos térmicos. Os grupos experimentais foram: G1: sem tratamento; G2: pré-aquecimento à 68°C com dispositivo Calset; G3: pós-aquecimento em forno MP-130, 10°C/min até 140°C e G4: pré e pós-aquecimento. Depois da fotoativação, foram realizadas análise termogravimétrica e calorimetria exploratória diferencial (DSC) para determinar a estabilidade térmica dos compósitos, temperatura de transição vítrea (Tg), pico de exotermia e calor liberado após fotoativação. O ensaio de resistência à flexão foi realizado 24h após armazenagem dos corpos de prova em água destilada a 37°C. A análise do grau de conversão foi realizada com espectrofotômetro FT-Raman. Análise de variância e teste de Tukey (p<0,05) demonstraram que os compósitos FT e ET têm 25,8% e 23,7% de matriz orgânica em peso respectivamente e estabilidade térmica até 200°C. O pico exotérmico do G2 (63,5°C) é maior do que G1 (60,9°C). Os grupos G3 e G4 não apresentaram pico exotérmico e calor liberado. A tg é de aproximadamente 160,5°C para o FT e de 161,4°C para o ET. A resistência à flexão (MPa) e grau de conversão(%) foram respectivamente: G1 (149,1; 60)=G2(152,5; 56,4) e G3 (170,5; 72,6)=G4 (178,2; 71,8) . O compósito ET apresenta maior grau de conversão que FT. À partir da caracterização térmica foi possível observar que o pré-aquecimento em condições não isotérmicas não promoveu maior conversão e resistência à flexão. O tratamento térmico deve ser realizado acima de 160°C. / The aim of this study was to perform the thermal characterization,flexural strength and degree of conversion of two composites, a nanofilled (Filtek Supreme XT: FT) and a microhybrid (Esthet X: ET), shade A2, submitted to different thermal treatments. Methods: The experimental groups were: G1: no treatment; G2: preheated at 68°C with Calset device; G3: post-heated in a MP-130 (EDG) oven, 10 °C/min up to reach 140°C for 20min and G4: pre and post-heated. After curing, thermogravimetric analysis and differential scanning calorimetry (DSC) were carried to determine thermal stability of the composites, glass transition temperature (Tg), exothermic peak and heat releasing after photoactivation. Three-point flexural test was performed after stored in water at 37oC for 24 hours. Degree of conversion was determined using a spectrophotometer FT-Raman. Analysis of variance and Tukey\'s test (P<0.05) showed that FT and ET composites have 25.8% and 23.7% of organic matrix in weight respectively and thermal stability at 200°C. The G2 group release more heat (9.9J/g) than G1 (7.3J/g). Exothermic peak of G2 group (63.5°C) is higher than G1 (60.9°C). G3 and G4 groups did not show exothermic peak and heat releasing. ET composite of G2 group presented the highest exothermic peak (65.2°C) of all groups. The Tg is nearly 160.5°C to FT and 161.4°C to ET. Flexural strength (MPa) and degree of conversion (%) were respectively: G1(149.1; 60) = G2(152.5; 56.4) and G3 (170.5; 72.6) = G4 (178,2; 71,8). ET composite presented higher conversion values than FT. Pre-heating at non-isothermal conditions does not promote higher conversion and flexural strength. Heat treatment after cure can be carried through 160°C.

Este trabalho descreve a construção e testes de um calorímetro eletromagnético de criptônio liquido utilizando uma geometria projetiva na forma de acordeom para os absorvedores eletrodos. Testes do calorímetro com feixes de elétrons de energia entre 20 e 200 GeV, realizados na linha de feixe H4 do acelerador SPS do CERN, apontam para uma resolução em energia de 5.63/(E[GeV] )0.26)% e uma não linearidade integral máxima de +/- 0,3%. O calorímetro é dividido longitudinalmente em 3 seções, sendo a primeira altamente segmentada transversalmente em uma direção, formando um pre-shower integrado ao calorímetro. A posição de incidência da partícula no calorímetro pode ser determinada com uma resolução de 2.16/(E[GeV] )0,19 7.93 / E[GeV])mm no pre-shower e o ângulo de incidência da partícula (utilizando a informação de posição das outras seções) com uma resolução de (32.01/(E[GeV] ) 0.95 44 /E[GeV])mrad. Os sinais do calorímetro permitem a realização de medidas de tempo com uma resolução de 290ps com elétrons de 120 GeV. Também são apresentados e discutidos o efeito da presença de material inativo na resolução em energia, a resposta do calorímetro a múons e os resultados da comparação entre simulações de Monte Carlo com medidas feitas com feixes de partículas. Por fim, é feito um estudo comparativo da resolução em energia obtida com este calorímetro e com um calorímetro de argônio liquido de geometria semelhante. / This work describes the construction and tests of a liquid krypton electromagnetic calorimeter employing an accordion shaped projective geometry for the absorbers and electrodes. The tests of the calorimeter using electron beams of energy ranging from 20 to 200 GeV and a muon beam of 225 GeV, from the H4 beam line of the CERN SPS, showed an energy resolution of 5,63/(E[GeV] ) 0.26)% and a maximum integral non-linearity of +/- 0.3%. the calorimeter is divided longitudinally in 3 sections, the first one being highly segmented on the transversal direction, operating as a pre-shower integrated on the calorimeter. The position of incidence for a particle in the calorimeter can be determined with a resolution of (2.16/E[GeV] 0.19 7.93 /E[GeV])mm on the pre-shower and the angle of incidence (using the position information from all sections) with a resolution of (32.01/E[GeV] 0.95 44 /E[GeV])mrad. The calorimeter signal allow timing measurements to be performed with a resolution of 290ps for 120 GeV electrons. It is also presented and discussed the energy resolution behavior in the presence of dead material in front of the calorimeter, the calorimeter response to muons and the comparison between the results from data and Monte Carlo calculations. Finally, it is presented a comparative study between the energy resolution of this calorimeter and a liquid argon calorimeter employing a similar accordion shaped geometry.

A galectina-4 humana (HGal-4), pertencente à família das galectinas, possui dois domínios de reconhecimento de carboidratos (CRDs) com alta afinidade para &beta;-galactosídeos e se encontra amplamente distribuída em células normais e neoplásicas de diferentes organismos. Suas funções snglobam uma grande variedade de eventos celulares, tais como processos inflamatórios, neoplásicos, progressão tumoral e metástase. Entretanto, muitas perguntas sobre suas interações com diferentes carboidratos, a especificidade destas interações e o papel específico das galectinas permanecem ainda sem resposta. No presente trabalho, propomos a investigação das interações galectina-glicano da galectina-4 humana e de seus domínios CRDs independentes (CRD-I e CRD-II) através de um conjunto de métodos biofísicos. Através do método de dicroísmo circular (CD), usando várias regiões espectrais, e fluorescência fomos capazes de entender mudanças ocorrentes na estrutura secundária e terciária das protéinas quando da interação com lactose/sacarose. Estes dados, juntamente com testes de hemaglutinação, mostraram que a glectina-4 e os CRDs respondem de forma distinta à ligação com açúcar. Por diferentes técnicas (fluorescência, ITC e MST) determinamos as constantes de dissociação para os domínios CRDs (Kd ~0,5 mM) e para HGal-4 e, de forma qualitativa, os valores obtidos indicaram possíveis estados oligoméricos dessas proteínas. A investigação da interação proteína-membrana da HGal-4 foi feita, primeiramente, com miméticos de membranas e monitorada pela técnica de RPE em crescente complexidade de composição de tais miméticos, indo desde composições mais simples, passando por lipid rafts na presença de diferentes glicolipídeos (GM1, LPS) e chegando-se à interação com células tumorais (U87MG, T98G e HT-29). Tais experimentos mostraram que galectina-4 reconhece e se liga naqueles modelos onde existem glicanos complexos na superfície. Investigamos também a participação de HGal-4 endógena e exógena no tratamento quimioterápico de células tumorais e verificamos um papel importante de HGal-4 para células HT-29. Finalizando esta tese, apresentamos o trabalho realizado em um ano de estágio na University of Oxford, durante o qual, investigamos a estrutura da região C-terminal de um receptor da família GPCR, qual seja o receptor de neurotensina NTS1. Aqui, mais uma vez, foi empregada a técnica de RPE que aliada à produção/marcação de mutantes do receptor, permitiu determinar que a hélice H8 se estabiliza quando em proteolipossomos. / Human galectin-4 (HGal-4), a member of the galectin family, contains two carbohydrate recognition domains (CRDs) with high affinity for &beta;-galactosides and is widely distributed in normal and neoplastic cells of different organisms. Its functions include a wide variety of cellular events such as inflammation, cancer, cell adhesion, tumor progression and metastasis. However, many questions about their interactions with different carbohydrates, the specificity of these interactions and the specific role of galectins remain unanswered. In this study, we propose the investigation of galectin-glycan interactions of human galectin-4 and its independent CRDs (CRD-I and-II) through a combination of biophysical methods. From circular dichroism (CD), measured in different spectral ranges, and fluorescence experiments we were able to understand changes in secondary and terciary structure of the protein while interacting with lactose/sucrose. These results along with hemagglutination assays showed that galectin-4 and its CRDs respond differently to sugar binding. From fluorescence, ITC and MST measurements we determined the dissociation constants for the CRDs (Kd ~0.5 mM) and for HGal-4. These values qualitatively indicated the formation of potential oligomers of CRDs and of HGal-4. The investigation of the HGal-4 interaction with membranes was firstly performed using mimetic membranes and monitored by EPR spectroscopy. The composition of the mimetic membranes was gradually increased so that to span simple compositions (such as DMPC), passing by lipid rafts in the presence of different glycolipds (GM1, LPS) up to interactions with tumor cells (U87MG, T98G e HT-29). These experiments showed that galectin-4 recognizes and binds to membrane models constituted by complex glycans on their surface. We also investigated the involvement of endogenous and exogenous HGal-4 in chemotherapies of tumor cells and found an important role of HGal-4 in the case of HT-29 cells. At last, we presented the work done in an one-year internship at the University of Oxford, during which we investigated the C-terminal region of the GPCR family receptor, the neurotensin receptor NTS1. Here, we used once again the EPR technique combined with the production/spin-labelling of mutants of the receptors, and determined that helix H8 was stabilized upon receptor reconstitution in proteolipossomes.