Master of Science in Medicine / Rationale: Asthma management aims to achieve optimal control on the minimal effective dose of medication. We assessed the effectiveness of two algorithms to guide ICS dose in well-controlled patients on ICS+LABA in a double-blind study, comparing dose adjustment guided by exhaled nitric oxide (eNO) to clinical care algorithm(CCA) based on symptoms and lung function. Methods: We randomised non-smoking adult asthmatics on minimum FP dose 100μgs daily +LABA to ICS adjustment using eNO or CCA, assessed over 5 visits during 8 months treatment. Primary endpoints were asthma-free days and asthma related quality of life (QOL). Analysis was by mixed model regression and generalised estimating equations with log link. Results: 69 subjects were randomised (eNO:34, CCA:35) and 58 completed the study. At baseline mean FEV1 was 94% pred., mean eNO (200ml/sec) 7.1 ppb, median ACQ6 score 0.33. Median ICS dose was 500 μg (IQR 100-500) at baseline and 100 μg on both eNO (IQR 100-200) and CCA arms (IQR 100–100) at end of study. There were no significant differences between eNO and CCA groups in asthma-free days (RR=0.92, 95% CI 0.8–1.01), AQL (RRAQL<median = 0.95, 95% CI 0.8–1.1) or exacerbation-free days (HR = 1.03, 95%CI 0.6–1.7). Neither clinic FEV1 (overall mean difference FEV1 % pred. -0.24%, 95% CI -2.2–1.7) nor a.m. PEF (mean difference 1.94 L/min (95% CI -2.9–6.8) were significantly different. Similar proportions of subjects were treated for ≥1 exacerbation (eNO: 50%, 95% CI 32.1–67.9; CCA: 60%, 95% CI 43.9–76.2). Conclusion: Substantial reductions in ICS doses were achieved in well controlled asthmatics on ICS+LABA, with no significant differences in outcomes between eNO or clinically based algorithms.

Master of Science in Medicine / Rationale: Asthma management aims to achieve optimal control on the minimal effective dose of medication. We assessed the effectiveness of two algorithms to guide ICS dose in well-controlled patients on ICS+LABA in a double-blind study, comparing dose adjustment guided by exhaled nitric oxide (eNO) to clinical care algorithm(CCA) based on symptoms and lung function. Methods: We randomised non-smoking adult asthmatics on minimum FP dose 100μgs daily +LABA to ICS adjustment using eNO or CCA, assessed over 5 visits during 8 months treatment. Primary endpoints were asthma-free days and asthma related quality of life (QOL). Analysis was by mixed model regression and generalised estimating equations with log link. Results: 69 subjects were randomised (eNO:34, CCA:35) and 58 completed the study. At baseline mean FEV1 was 94% pred., mean eNO (200ml/sec) 7.1 ppb, median ACQ6 score 0.33. Median ICS dose was 500 μg (IQR 100-500) at baseline and 100 μg on both eNO (IQR 100-200) and CCA arms (IQR 100–100) at end of study. There were no significant differences between eNO and CCA groups in asthma-free days (RR=0.92, 95% CI 0.8–1.01), AQL (RRAQL<median = 0.95, 95% CI 0.8–1.1) or exacerbation-free days (HR = 1.03, 95%CI 0.6–1.7). Neither clinic FEV1 (overall mean difference FEV1 % pred. -0.24%, 95% CI -2.2–1.7) nor a.m. PEF (mean difference 1.94 L/min (95% CI -2.9–6.8) were significantly different. Similar proportions of subjects were treated for ≥1 exacerbation (eNO: 50%, 95% CI 32.1–67.9; CCA: 60%, 95% CI 43.9–76.2). Conclusion: Substantial reductions in ICS doses were achieved in well controlled asthmatics on ICS+LABA, with no significant differences in outcomes between eNO or clinically based algorithms.

A series of phosphotripeptide ligands for the Grb2 SH2 domain was designed and synthesized, each of which derived from the minimal consensus sequence required for binding: Ac-pYXN. The binding affinity and related thermodynamic parameters were determined by isothermal titration calorimetry. Both the size and connectivity of the side-chain was varied. The consequences of incorporating α,α-disubstitution at the pY+1 residue on binding thermodynamics were evaluated, as were the effects of constraining the side-chains in a ring. The series was evaluated from a number of perspectives: (1) increasing size of the pY+1 residue by utilizing various amino acid types: monoalkyl, dialkyl, or cycloalkyl; (2) comparisons between ligands with the same number of carbons (scission control); and (3) by comparing ligands incorporating cyclic pY+1 residues with those incorporating α,α-dialkyl residues with one fewer methylene group (excision control). Inconsistencies in the thermodynamic consequence of constraining the backbone were observed within this set of ligands, which reveal the limitations of our understanding of protein-ligand interactions. Aspects of both the classical and non-classical hydrophobic effect were observed, but the occurance of one over the other could not be explained. / text

Cancer is a worldwide health problem and despite advances in traditional treatments i.e. surgery, chemotherapy and radiotherapy, the cure rate remains disappointing for some cancers. Different novel therapeutic strategies are being developed. In this thesis two nontraditional cancer therapy approaches are studied: gene therapy using viral vectors and antitumour vaccination with dendritic cell - tumour cell (DC/TC) hybrids. We have developed a novel ELISPOT titration method for viral vectors that is based on the actual expression of the transgene in target cells. This method was developed with recombinant parvovirus MVM-IL2, but it should be adaptable for other vectors carrying expression cassettes for secreted transgene products for which antibodies are available. The ELISPOT titration method allows for faster and better quantification of transducing units present in vector stocks as opposed to titration by in situ hybridisation (annexe I). The MVMIL2 vector has shown an anti-tumour effect against melanoma in an immunocompetent mouse model (annexe IV). Previous work concerns photodynamic inactivation of adenoviral vectors for biosafety and an in vivo study in which a synergistic effect of antiangiogenesis gene therapy combined with radiotherapy could be shown (annexes V and VI). DC/TC hybrids have been proposed as cancer vaccines for their simultaneous expression of antigen presentation machinery and tumour associated antigens. Hybrids are classically generated by polyethylene glycol (PEG) or electrofusion. These methods however require special skills and equipment and cause rather high cell lethality. Fusion via the expression of viral fusogenic membrane glycoproteins (FMG), such as the vesicular stomatitis virus-G (VSV-G) (annexe III) or the Gibbon ape Leukemia Virus (GaLV) FMG, have recently been described. We have mainly focussed on the latter. Transduction of cells with GaLV-FMG proved to be a limiting step for an efficient generation of hybrids. On the other hand, constitutive expression of GaLV-FMG leads to lethal syncytia formation in human cells. Therefore we developed a novel fusion strategy for the generation of DC/TC cell hybrids that involves the use of a non-human fusogenic cell line that constitutively expresses the GaLV-FMG. With this method we were able to generate reproducible yields of DC/TC triparental hybrids. The formation of tri-parental hybrids via the fusogenic cell line is an interesting alternative to existing DC/TC fusion methods because of its simplicity and its flexibility in the choice of fusion partners, i.e. autologous or allogeneic DCs and tumour cells. Moreover, the tri-parent hybrid system offers the possibility to further enhance the immune response by the addition of transgenes that code for immuno-modulating factors to the fusogenic cell line (annexe II).

The preparation of layered double hydroxides via titration with sodium hydroxide was thoroughly investigated for a number of M(II)/M(III) combinations. These titration curves were examined and used to calculate nominal solubility product constants and other thermodynamic quantities for the various LDH chloride systems.

Orientador: José Paschoal Batistuti / Banca: Fernando Luis Fertonani / Banca: Valeria Monterio da Silva Eleutério / Resumo: O presente trabalho consiste no desenvolvimento de um método analítico para determinação de biotina, utilizando-se a potenciometria indireta. O objetivo é apresentar a validação de método analítico como um processo que estime a eficiência do método proposto na rotina do laboratório para garantia da qualidade total. É um método que envolve equipamento simples e pouco dispendioso como o potenciômetro, que possibilita medir com precisão o valor da concentração de biotina. O doseamento quantitativo de biotina baseia-se no estudo das reações oscilantes do analito perante as análises da titulação indireta utilizando potenciômetro automático. A concentração de biotina foi determinada com massa adicionada conhecida numa matriz que simula uma cápsula, contendo aerosil (1%), estearato de magnésio (1%), celulose microcristalina (20%), amido (40%) e lactose q.s.p. Os resultados mostraram um valor médio de biotina (massa adicionada de aproximadamente 25 mg) determinada de 99,4% com desvio-padrão de 0,0345. As condições experimentais como temperatura, vidraria e concentração dos reagentes foram otimizadas. Os parâmetros investigados no processo de validação para demonstrar o desempenho do método foram: especificidade, linearidade, intervalo, precisão tanto repetitividade, quanto intermediária, exatidão e robustez. O tratamento estatístico dos dados da validação do método analítico envolveu a determinação da média, do desvio padrão e do coeficiente de variação. Para obtenção da curva de calibração se fez necessária a determinação da equação da reta, regressão linear e coeficiente de correlação linear. Este método apresenta grande aplicabilidade em soluções turvas, fluorescentes, opacas ou coradas, ou quando não existem, ou não podem aplicar-se indicadores visuais apropriados. Há possibilidade de determinação... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The present work shows the development of an analytical method for the determination of biotin by using indirect potentiometry. The main goal is to present the validation of analytical methods as a process to estimate the efficiency of the proposed methodology in the laboratory routine for the guaranty of total quality. This method involves simple and low cost devices as the potentiometer, which allows the precise measurement of biotin concentration. The quantitative dosing of biotin is based on the study of the oscillating chemical reactions with the analyte by performing the analysis of the indirect titration with an automatic potentiometer. Biotin concentration was determined by adding a known mass to a matrix that simulates a capsule containing aerosol (1%), estearato de magnésio (1%), celulose microcristalina (20%), amido (40%) e lactose q.s.p. The results showed an average value for the determination of biotin (added mass was ca. 25 mg) of 99.4% with a standard deviation of 0.0345. The experimental conditions as temperature, glassware, and concentration of the chemicals were optimized. The investigated parameters of the validation procedure to demonstrate the performance of the method were: specificity, linearity, interval, precision (repeatability and intermediate), exactness and robustness. The statistical treatment of the data for the validation of the analytical method involved the determination of the average value, standard deviation, and the variation coefficient. In order to obtain the calibration curve, the line's equation, the linear regression and the coefficient of linear correlation were determined. This method shows great applicability for turbid, fluorescent, opaque or color solutions, or if an appropriate visual indicator is not available or cannot be applied. The method also shows the possibility for the determination of a sequence of equivalence... (Complete abstract click electronic access below) / Mestre

Photocopy of typescript. / Digitized by Kansas Correctional Industries

Vitamin C occurs in relatively high concentrations in fresh and processed fruits and vegetables but is found to a lesser extent in animal tissues and animal-derived products. Nearly 90 % of vitamin C in the human diet is obtained from fruits and vegetables but this can be indirect by way of commercially prepared fruit juices. These juices are often enriched with vitamin C which has been synthetically prepared. There is a wide range of such juices on the New Zealand market, and they are a significant source of dietary vitamin C for many in the population. The focus of this research is on the Keri range of juice products.The present study monitors the fate of vitamin C during storage of Keri juices up to the best-before date, and under a range of other storage and consumption situations. Two methods were adopted for determining ascorbic acid (AA, the chemical identity of vitamin C). These were the titrimetric method, which is based upon the reduction of the dye 2,6-dichlorophenolindophenol by AA in acidic solution, and liquid chromatography, which is used to separate AA from its immediate oxidation product dehydroascorbic acid. In the latter method these two analytes can be measured independently. The liquid chromatography was less successful than the simpler titrimetric method, so most of the work was done by titration. However, the concentration of dehydroascorbic acid, which has vitamin C activity in vivo, remained uncertain. Moreover, the titrimetric method could not be applied to juices with high purple anthocyanin concentrations, like blackcurrant, because the colour change at the titration end point could not be detected. pH adjustment to change colour was ineffective, and decolourisation with charcoal led to the rapid and complete destruction of AA. The concentration of AA in Keri juices at the time of manufacture were always much higher than claimed on the labels. Storage for up to nine months at room temperature resulted in a loss in AA of between 37 and 68 %, depending on the juice and exposure to fluorescent light. However, the time of storage was a much more dominant factor than light exposure. The kinetics of loss, straight lines, were most easily explained by an aerobic model of AA degradation from oxygen diffusing across the polyethylene tetraphthalate bottle wall. Overall, the label claims made were defensible in terms of the best-before date, because it took at least 100 days of storage before the AA concentration in the most susceptible juices fell below the claimed value. This is because these drinks are fast moving consumer goods and storage beyond 100 days is unlikely. (Nonetheless, the supplier (Keri Juice Company) has since adopted its new unitised method of formulating juice. This has resulted in an initially higher concentration of vitamin C as compared to the juices under investigation.) In the nine months storage experiment there was some evidence for the presence of dehydroascorbic acid in blackcurrant drinks, but not in another three juices. Pasteurisation during preparation of these drinks resulted in up to 7 % loss of AA, probably due to oxygen dissolved in water, and accelerated by heat of pasteurisation. Higher temperatures in later storage also accelerated losses. Progressive exposure of juice to air during simulated consumption of 3 L bottles over a week also accelerated losses. Finally, exposure to sunlight in a diurnal temperature environment accelerated losses five-fold higher than in total darkness. Filtration of ultraviolet light approximately halved the loss due to sunlight. Overall however, it can be concluded that AA in the Keri range of juices is very resistant to degradation of AA.

Atom transfer radical polymerisation (ATRP) technique was used to synthesize poly(methacrylic acid-block-methyl methacrylate) (P(MAA₁₀₂-b-MMA₁₀)) copolymer in order to study the aggregation behavior in aqueous solution over the course of neutralization. A combination of static and dynamic light scattering (SLS, DLS) and potentiometric titration techniques were used to investigate the size and shape of the micelle at various degrees of neutralization. The hydrodynamic radius (Rh) determined from dynamic light scattering increases from ~26nm (for unneutralized) to ~42nm (for completely neutralized sample). Both potentiometric and laser light scattering studies indicate the formation of a core shell micelle. The weighted average molecular weights of the polymer and micelle are 1.18x10⁴ and 2.25 x 10⁵ g/mol respectively, which suggests that the aggregation number of the micelle is ~20. / Singapore-MIT Alliance (SMA)

Arbetet har mestadels utförts i kokeriet på Domsjö Fabriker i syfte att kartlägga hur processförhållandena hänger samman med koncentrationen sulfatjoner. Detta för att lättare kunna undvika förhållanden i processen som innebär mycket höga värden. Arbetet ingår i examen för högskoleutbildningen: processoperatör vid Umeå Universitet.Vid Domsjö Fabriker har tidigare liknande mätningar gjorts i kokeriet och det har konstaterats att koncentrationen sulfatjoner hänger samman med pH och tid. Syftet är att djupare kartlägga hur processförhållandena hänger samman för att lättare kunna undvika förhållanden som innebär höga koncentrationer sulfatjoner. Målet med examensarbetet och dess undersökning är att sammanställa resultat som visar hur och när under koket som sulfatjoner bildas i stora mängder i kokeriet. Förfarandet av undersökningen har bestått i att prov tas ut i provserier om fyra provtillfällen per kok, fryses in och slutligen analyseras genom titrering och pH-mätning.Efter totalt 22 stycken provserier om fyra prov från de olika kokarna konstaterades att sulfatkoncentrationen är högst i slutet av nedgasningssteget. Det första provtillfället hade de högsta pH-värdena men också de lägsta koncentrationerna sulfat. Inget av dessa värden sticker ur och anses onormalt på något sätt. Om sulfatkoncentrationen påverkas av pH-värdet går inte att fastställa av detta arbete, då det inte är de lägsta pH-värdena som genererar de högsta sulfatkoncentrationerna utifrån resultatet i undersökningen. / This work has been performed in the digester at Domsjö Fabriker in order to clarify the concentration of sulphate-ions comparing to different driving-conditions in the process. The reason to this is to understand and be able to avoid conditions in the digester process that means very high concentrations of sulphate-ions. The work is included in the examination of higher education: process operator at Umeå University.At Domsjö Fabriker similar studies have been done in the digester and from that it has been found out that the concentration of sulphate-ions is connected to pH and over time. The purpose is to approve the understanding of the process conditions to avoid extremely high concentration of sulphate-ions. The mission with this work and the measurements included in the work is to put together results that show when the concentration of sulphate-ions is high. The procedure of the study has consisted in taking samples in series of four, frozen the samples and finally analyzing them by titration and pH-measurements.After 22 evaluated trial-series á four samples each from the boilers it was concluded that the concentration of sulphate-ions was the greatest in the end of this process-step. The pH was upper and the concentration of sulphate lower in the first sample. None of the data is abnormal in any way, and they are all connected and relevant to each other. In this experiment it is not possible to state if the concentration of sulphate is affected by the pH or not.