Orientador: Jose Pedrazzoli Junior / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-12T03:40:36Z (GMT). No. of bitstreams: 1 Morita_MilenaRodrigues_M.pdf: 4296553 bytes, checksum: 165726f88ad105c8817c9cc33149c67a (MD5) Previous issue date: 2008 / Resumo: Objetivo: Desenvolver e validar um método analítico para quantificação de levocetirizina em plasma humano. Além disso, a biodisponibilidade relativa de uma formulação contendo 5 mg de dicloridrato de levocetirizina (formulação teste e formulação referência produzida por Farmalab Indústrias Químicas e Farmacêuticas Ltda.) foi avaliada em trinta e seis voluntários sadios de ambos os sexos. Método: O plano de estudo utilizado foi aberto, randomizado, cruzado com um intervalo de washout de 7 dias. As amostras de plasma foram obtidas por um período de 48 horas. As concentrações plasmáticas de levocetirizina foram analisadas por cromatografia líquida de alta eficiência acoplada à espectrometria de massas (HPLC-MS/MS), com modo de ionização electrospray positivo usando um monitoramento de reação simples (SRM). O método analítico foi validado considerando os seguintes parâmetros: especificidade, linearidade, precisão, exatidão, recuperação e estabilidades de acordo com a RE n°899/03 (ANVISA). Das curvas de concentração plasmática versus o tempo para levocetirizina, os seguintes parâmetros farmacocinéticos foram obtidos: ASC0-48h, ASC0-inf e Cmax. Resultados: A curva de calibração foi linear de 0,5 a 500,0 ng/mL. O método obteve uma corrida cromatográfica de 2,0 minutos usando uma coluna POLARIS C18 (3 µm, 50 mm x 2,0 mm). A precisão inter-corrida dos controles de qualidade foi 4,76% (CQB 1,5 ng/mL), 7,14% (CQM 200,0 ng/mL) e 3,78% (CQA 400,0 ng/mL). A exatidão inter-corrida dos controles de qualidade acima mencionados foi de 98,00%, 99.63% e 97,84%, respectivamente. A razão das médias geométricas dos dois medicamentos (teste e referência) e o intervalo de confiança (IC) foram respectivamente: 101,11% (90% IC= 97,42% - 104,93%) para ASC0-48h, 101,17% (90% IC= 97,49% - 104,99%) para ASC0-inf, 99,43% (90% IC= 94,29% - 104,86%) para Cmax. Conclusão: O método analítico mostrou-se preciso, exato e rápido para quantificação de levocetirizina em plasma humano. Desde que os IC 90% para ASC0-48h, ASC0-inf e Cmax apresentaram-se dentro do intervalo de 80-125% proposto pela ANVISA, foi concluído que a formulação teste Levocetirizina 5 mg é bioequivalente a formulação Zyxem® fabricada pelo laboratório Farmalab Indústrias Químicas e Farmacêuticas Ltda. / Abstract: Objective: To develop and validate an analytical method for levocetirizina quantification in human plasma. Moreover, it was evaluated the relative bioavailability of a levocetirizine dichloridrate 5 mg tablet formulation (test formulation vs reference formulation from Farmalab Ltda.) in 36 healthy volunteers of both sexes. Methods: The study was conducted using an open, randomized, two-period crossover design with 7 days washout period between doses. Plasma samples were obtained over a 48 h period. Plasma levocetirizine concentrations were analyzed by liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) with positive ion electrospray ionization using single reaction monitoring (SRM). The analytical method was validated considering specificity, linearity, precision, accuracy, recovery and stabilities parameters according to RE n°899/03 (ANVISA). From the levocetirizine plasma concentration vs time curves, the following pharmacokinetic parameters were obtained: AUC0-48h, AUC0-inf e Cmax. Results: The calibration curve was linear over de range from 0,5 to 500,0 ng/mL. The method chromatographic run was 2,0 minutes using a POLARIS C18 column (3 µm, 50 mm x 2,0 mm). The between-run precision of quality controls was 4,76% (CQB 1,5 ng/mL), 7,14% (CQM 200,0 ng/mL) e 3,78% (CQA 400,0 ng/mL). The between-run accuracy for the above mentioned quality controls was 98,00%, 99.63% e 97,84% respectively.The limit of quantification was 0.5 ng/mL. The geometric mean ratio of both formulations (test and reference) and respective 90% confidence interval (CI) were: 101,11% (90% CI= 97,42% - 104,93%) for AUC0-48h, 101,17% (90% CI= 97,49% - 104,99%) for AUC0-inf, 99,43% (90% CI= 94,29% - 104,86%) for Cmax. Conclusion: The analytical method has proven to be precise, accurate and fast to levocetirizina quantification in human plasma. Since the 90% CI for AUC0-48h, AUC0-inf and Cmax ratios were within the 80-125% interval proposed by the ANVISA, it was concluded that levocetirizine formulation elaborated by Eurofarma Laboratórios Ltda. is bioquivalent to Zyxem® formulation. / Mestrado / Mestre em Farmacologia

La maladie de Fabry est une maladie de surcharge lysosomale liée au chromosome X. Elle est causée par une mutation au niveau du gène GLA qui provoque un déficit de l’enzyme α-galactosidase A. Elle entraîne une accumulation de glycosphingolipides tels le globotriaosylcéramide (Gb3), le globotriaosylsphingosine (lyso-Gb3) et galabiosylcéramide (Ga2) ainsi que leurs isoformes/analogues respectifs au niveau des tissus et des liquides biologiques des patients. Les symptômes peuvent se présenter de manière très variable soit par de l’acroparesthésie, des troubles ophtalmologiques, des angiokératomes, des troubles cardiaques, rénaux ou encore neurologiques. Les connaissances au niveau physiopathologique de la maladie de Fabry sont à ce jour, toujours déficientes. Les biomarqueurs analysés ne permettent pas de prédire la sévérité et la progression de la maladie. Afin d’apporter des connaissances plus approfondies à ce niveau, l’objectif principal de cette étude était d’analyser les biomarqueurs au niveau cellulaire pour des patients atteints de la maladie. Les objectifs secondaires étaient: 1) de développer et de valider une méthode en spectrométrie de masse en tandem pour la quantification relative et simultanée de certains biomarqueurs susmentionnés dans les leucocytes, les lymphocytes B et les monocytes de patients Fabry et contrôles sains; 2) d’évaluer les biomarqueurs dans le total des leucocytes, les lymphocytes B, les monocytes, le plasma et l’urine chez les patients Fabry et les contrôles sains appariés selon le sexe et l’âge; et 3) d’établir des corrélations entre la quantité relative de ces biomarqueurs et le génotype des patients traités et non traités. Les résultats de cette étude démontrent qu’il n’y a pas de changements significatifs dans la distribution des groupes d’isoformes/analogues du Gb3 selon le type cellulaire pour les patients et les contrôles. La découverte d’isoformes/analogues méthylés du Gb3 suggère un processus de méthylation qui se produit directement au niveau des cellules sanguines et particulièrement pour les lymphocytes B et les monocytes. Des corrélations face à la quantité de biomarqueurs et le génotype ont été établies. Ces résultats pourraient permettre une meilleure compréhension des processus biochimiques reliés à l’accumulation du Gb3 dans les cellules sanguines.

Magister Scientiae - MSc / The objective of the study was to conduct an internal validation on miniSTR systems and apply it to cases received from the South African Missing Persons Task Team (SAMPTT). This was prompted by the fact that miniSTR systems have been shown to out perform some of the commercial kits available in the time of the study and provide an alternative to mtDNA when analysing degraded DNA from skeletal remains and that the DNA extracted from skeletal remains received from the SAMPTT would be degraded due to the remains generally being fragmented or charred and buried for many years. The miniSTR loci chosen for validation comprised the Combined DNA Index System (CODIS) thirteen core loci and were arranged into four triplexes and one uniplex. / South Africa

>Magister Scientiae - MSc / In this study the 10 Y-specific loci of the University of the Western Cape (DYS710, DYS518 385a/b, DYS644, DYS612, DYS626, DYS504, DYS447, DYS447, and DYS481) were analysed in 492 individuals from South African population groups. Four different populations namely; Zulu, Coloured, Afrikaner and Asian Indian were sampled. A total of 488 haplotypes were observed, 412 of which were unique. Haplotype diversity was 0.9981. Gene Diversity values ranged from 0.8075 for DYS447 to 0.9209 for DYS710. The discriminatory capacity was 0.9106 which is high. The study showed that the University of the Western Cape 10 locus is a powerful discrimination tool for routine forensic applications and could be used in genealogical investigations as compared to other commercial kits when used on the South African populations (Zulu, Coloured, Afrikaner and Asian Indian) considering its high discriminatory capacity. This data will be used for the establishment of a Y-STR DNA databases for South African population which would aid law enforcement authorities in the investigation and resolution of crimes AMOVA computed using haplotype frequencies showed that when male haplotypes from the four different populations were compared, 0.22 % of the total genetic variation was due to the variability among populations and 99.78 % of the total variation is found within populations. However AMOVA computed using distance matrix showed that 5.97 % of the total variation was due to variability among populations and 94.07 % of the total variation is found within populations. Genetic substructure was found among the four studied South African population groups. All the six population pairwise comparisons using AMOVA were significant .Therefore Y-STRs are very useful in comparing closely related populations. It should be noted that their utility for evolutionary purposes, they need to be combined more stable Y-DNA markers such as single nucleotide polymorphisms (SNPs). Factorial Correspondence Analysis (FCA) showed that the Coloured population has large genetic contribution from Afrikaner population and lesser contribution from the Zulu and Asian Indian population groups. / National Research Foundation (NFR)

>Magister Scientiae - MSc / The primary objective of this Masters project was to investigate the maternal ancient substructure of the Griqua population in South Africa. Genetic ancestry was determined by investigating ancestry informative single nucleotide polymorphisms. These are located in the control region of the mitochondrial genome. The auxiliary aim was to test the validity of the UWC 10plex system in relation to a sample group of Griqua males. This short tandem repeat multiplex targets specific mutations confined to paternal lineages. The Khoi Khoi or Hottentots were the first inhabitants in the Cape. Indigenous Khoi Khoi female slaves had offspring with the European settlers in the 1800s which resulted in the Griqua population group. The incorporated European paternal ancestry is what set the Griqua apart from the native population groups at that time. Colonisation events from the mid-17th to 19th Century and the apartheid regime resulted in land dispossession of the native population and an extensively mixed gene pool in South Africa. One hundred and seventy six (N=176) male and female Griqua people were collectively sampled in Kokstad (2012), Vredendal (2012 and 2013) and at the Griqua National Conference in Ratelgat (2013). All 176 samples were analysed using mtDNA control region Sanger sequencing. The sample group (N=176) was separated based on birthplace (Origin sample group and post-colonial sample group). The origin sample group consists of individuals whose ancestors were not part of the Griqua Trek to Northern regions of South Africa and were less likely to be exposed to colonial influences. Mutations within the hypervariable segments of the mtDNA control region were used to infer haplogroups with geographic-specific population data. In this way one can plot the extent of ancient Khoisan (L0d) and Bantu influences (L1-L5) as well as the influence of East (M, A, B, E) and West (N, R, J, H) Eurasian haplogroups in the maternal ancestry of the Griqua population group. The origin sample group showed 91% African ancestry (76.8% L0d) while the post-colonial group had 78% African ancestry (60% L0d). The origin sample group had 2% East Eurasian and 7% West Eurasian ancestry, while the post-colonial group contained 20% Eurasian ancestry. There is greater admixture in the post-colonial group which can be attributed to the integration of surrounding populations during settlement periods in parts of the Northern Cape and KwaZulu-Natal. The UWC 10plex STR kit was tested to see if it could discriminate between male individuals of this admixed sample group (N=91 males). The markers for this multiplex were selected according to their ability to differentiate between individuals of African descent. It proved to be a viable Y chromosome short tandem repeat testing tool, displaying a statistically significant discrimination capacity value of 0.966 and only having 3 shared haplotypes in the sample group of 91 Griqua males. / National Research Foundation (NRF)

L'objectif de ce travail est de développer un système catalytique hétérogène pour l'époxydation en utilisant directement O2 de l'air plutôt que H2O2 commercial. La stratégie adoptée a été de coupler la production in situ de H2O2, catalysée par la glucose oxydase (GOx), avec un catalyseur à base de Ti. La GOx a été immobilisée de manière covalente sur une mousse silicique mesocellulaire (MCF) amino-fonctionnalisée puis la stabilité thermique et aux solvants organiques de MCF-NH2-GOx a été étudiée. L'approche visant à ancrer Ti puis la GOx sur le même support n’a pas abouti à un catalyseur tandem efficace du fait d'un recouvrement de Ti par –NH2. L’hydrophilie de MCF apparaît, de plus, défavoriser l'oxydation d'alcènes organosolubles. Une option a consisté à utiliser la zéolithe TS-1 hydrophobe et réputée fonctionner en milieu aqueux mais dont les micropores ne peuvent loger la GOx. Cette dernière, associée à MCF-NH2-GOx en mélange mécanique, s’est montrée performante pour l'oxydation du cyclohexène dans MeOH/tampon acétate 50:50 à 35°C (rendement de 50% en époxyde et ses dérivés). D’encore meilleurs résultats ont été obtenus pour le prop-2-ène-1-ol en milieu aqueux à 40°C (rendement de 87% en glycérol). L’attaque basique de TS-1 crée une porosité suffisante pour loger la GOx, mais endommage son activité. En revanche, le recouvrement de MCF par un film de TS-1 a eu un effet bénéfique sur l’oxydation du prop-2-ène-1-ol dans l’eau par H2O2. Enfin, la porphyrine Mn-TCPP s’est montrée efficace comme catalyseur d'oxydation en tandem avec la GOx en solution mais, en cas d’immobilisation sur le support silicique MCF, la formation d’un précipité inhibe son activité. / The objective of this work was to develop an heterogeneous catalyst system supplied by dioxygen, rather than commercial H2O2, in order to carry out epoxidation reactions. Our strategy was to couple the in situ production of H2O2, catalyzed by glucose oxidase (GOx), with a Ti-based catalyst. The enzyme was covalently grafted onto a silicic mesocellular foam (MCF) functionalized by aminopropyle groups, then the thermal stability and behavior in organic solvents of the resulting material were investigated. The approach aiming at anchoring Ti, then GOx on the same support did not result in an effective tandem catalyst because of a too high –NH2 surface coverage. Hydrophilicity of MCF makes the oxidation of organosoluble alkenes unefficient. An alternative approach consisted in using the hydrophobic TS-1 zeolite known to operate in aqueous medium but whose micropores do not allow GOx hosting. However, TS-1 combined in a mechanical mixture with GOX immobilized on MCF turned out to be effective for the oxidation of cyclohexene in MeOH/acetate buffer 50:50 at 35°C (50% yield of epoxide and its derivatives). Even better performances were obtained for prop-2-ene-1-ol oxidation in aqueous medium at 40°C (87 % yield of glycerol). The basic attack of TS-1 has created mesoporosity to host GOx but damaged active Ti sites. On the other hand, TS-1 coated MCF appeared to be a good option having a beneficial effect on the oxidation of prop-2-en-1-ol in water by H2O2. Finally, a manganese porphyrin, Mn-TCPP, was also tested successfully as alkene oxidation catalyst in combination with GOx but, in case of immobilization, the presence of the silicate support lead to a deactivated catalyst.

Peptide identification using tandem mass spectrometry depends on matching the observed spectrum with the theoretical spectrum. The raw data from tandem mass spectrometry, however, is often not optimal because it may contain noise or measurement errors. Denoising this data can improve alignment between observed and theoretical spectra and reduce the number of peaks. The method used by Lewis et. al (2018) uses a combined constant and moving threshold to denoise spectra. We compare the effects of using the standard preprocessing methods baseline removal, wavelet smoothing, and binning on spectra with Lewis et. al’s threshold method. We consider individual methods and combinations, using measures of distance from Lewis et. al's scoring function for comparison. Our findings showed that no single method provided better results than Lewis et. al's, but combining techniques with that of Lewis et. al's reduced the distance measurements and size of the data set for many peptides.

Diploma thesis includes design of bearings of steering joints of tandem vibratory pivot-steering roller Ammann AV 95-2, design of bearings of steering joints of tandem vibratory with new type of steering kinematics and calculation and comparison of contact pressures between drum and soil for different angles of turn of front and rear drum for different types of tandem vibratory rollers. Comparison of contact pressures is realised for pivot steering roller AV 95-2, articulated tandem roller AV 80 X, articulated tandem roller with modified configuration of steering-joints and tandem roller with new type of steering kinematics.

abstract: The emergence of perovskite and practical efficiency limit to silicon solar cells has opened door for perovskite and silicon based tandems with the possibility to achieve >30% efficiency. However, there are material and optical challenges that have to be overcome for the success of these tandems. In this work the aim is to understand and improve the light management issues in silicon and perovskite based tandems through comprehensive optical modeling and simulation of current state of the art tandems and by characterizing the optical properties of new top and bottom cell materials. Moreover, to propose practical solutions to mitigate some of the optical losses. Highest efficiency single-junction silicon and bottom silicon sub-cell in silicon based tandems employ monocrystalline silicon wafer textured with random pyramids. Therefore, the light trapping performance of random pyramids in silicon solar cells is established. An accurate three-dimensional height map of random pyramids is captured and ray-traced to record the angular distribution of light inside the wafer which shows random pyramids trap light as well as Lambertian scatterer. Second, the problem of front-surface reflectance common to all modules, planar solar cells and to silicon and perovskite based tandems is dealt. A nano-imprint lithography procedure is developed to fabricate polydimethylsiloxane (PDMS) scattering layer carrying random pyramids that effectively reduces the reflectance. Results show it increased the efficiency of planar semi-transparent perovskite solar cell by 10.6% relative. Next a detailed assessment of light-management in practical two-terminal perovskite/silicon and perovskite/perovskite tandems is performed to quantify reflectance, parasitic and light-trapping losses. For this first a methodology based on spectroscopic ellipsometry is developed to characterize new absorber materials employed in tandems. Characterized materials include wide-bandgap (CH3NH3I3, CsyFA1-yPb(BrxI1-x)3) and low-bandgap (Cs0.05FA0.5MA0.45(Pb0.5Sn0.5)I3) perovskites and wide-bandgap CdTe alloys (CdZnSeTe). Using this information rigorous optical modeling of two-terminal perovskite/silicon and perovskite/perovskite tandems with varying light management schemes is performed. Thus providing a guideline for further development. / Dissertation/Thesis / Doctoral Dissertation Electrical Engineering 2019

In this project, samples with a metal/semiconductor/metal structure were fabricated and investigated with the potential application as the interconnecting layer of a tandem solar cell in mind. Degenerately doped p-Si and n-Si were used as bottom (metal like) contacts, as Si represents one of the most common materials for the bottom cell of tandem devices. A transparent, wide bandgap semiconductor in the form of SnO₂ was investigated for the intermediate layer as it is a common choice for the selective back contact of top cells based on perovskites. However, atomic layer deposition (ALD) was used as an alternative to the typical solution based application of the SnO₂ layer. The top layer was simply chosen as a triple layer metal contact stack (Ni-Al-Ni) to provide for good contact with the SnO₂.The goal of the project was to study the electrical properties of the samples through I-V measurements and how the I-V characteristic depends on the oxide’s thickness under the possible influence of the contact areas. Three different thicknesses of the SnO2 layer were used for the p-Si sample: 50, 200 and 400 Å. For the n-Si samplesonly one thickness (400 Å) was studied. Using the diode equation, four parameterswere calculated (Jo, Rsh, Rs and n) for different measurements combing different contact configurations. The latter included measurements between the front and the back of the samples and measurements between contacts on the front with and/orwithout SnO2 layer. From the results, it was concluded that as the thickness of SnO₂ increases, the saturation current (Jo) decreases while both shunt resistance (Rsh) andseries resistance (Rs) increase. The ideality factor (n) neither depends significantly on effective area, nor on SnO2 thickness. The p-Si and n-Si samples show similar behavior in the case of 400 Å SnO2 thickness. The contact areas only appreciatively affect Jo, but it is not clear what lies behind this dependence. In all cases, the top contacts obtained major wear during measurements, reducing the number of trustworthy measurements that could be used on the smaller areas. The resistivity through the oxide layer was calculated to ρSnO₂ = 247±96 MΩ cm, which is higher than for SnO₂ deposited by other techniques, and too high for tandem cell application. Schottky barriers formed at the interfaces will typically limit the charge transport further.