Graduation date: 1993

Thesis (MSc)--University of Stellenbosch, 2007. / ENGLISH ABSTRACT: Wine is a complex medium. Wine aroma, flavour and colour are important quality factors, but these can be influenced by many factors, such as grape-derived compounds that exist as free volatiles and also as glycosidically bound. The chemical composition of wine is determined by factors such as grape variety, geographic position, viticulture condition, microbial ecology of the grape and the winemaking process. The varietals aroma is determined by both the volatile and the non-volatile compounds, such as monoterpenes, norisoprenoids and benzene derivatives, which are naturally present in the wine. Monoterpenes are very important in the flavour and aroma of grapes and wine. They can be found in grapes and wine either in the free, volatile and odorous form, or in the glycosidically-bound, non-volatile and non-odorous form. The ratio of glycosidically-bound compounds to free aroma compounds is very high in the Gewürztraminer, Muscat and Riesling cultivars in particular. The glycosidic bonds can be hydrolysed either by the acid method or by using enzymes. The acid method is disadvantageous because it can modify the monoterpenes, whereas enzymatic hydrolysis has the advantage of not modifying the aroma character. The enzyme method of breaking the glycosidic bonds occurs in two successive steps: initial separation of glucose from the terminal sugar by a hydrolase (a-L-arabinofuranosidase, a-L-rhamnosidase or β-apiosidase, depending on the aglycone moiety), followed by the breaking of the bond between the aglycone and glucose by β-glucosidase. The enzyme β-glucosidase can be obtained from many plant (Vitis vinifera), bacterial, yeast or fungal sources. Most of the enzymes produced by these sources are not functional under the winemaking conditions of low pH, low temperature, high glucose and high ethanol content. However, β-glucosidases from fungal origins, particularly from Aspergillus spp., are tolerant of winemaking conditions. The idea of using the β-glucosidase gene from the fungus Aspergillus kawachii (BGLA), which is linked to the cell wall and the free β-glucosidase, was to determine if anchoring the enzyme to the cell wall will increase the activity of the enzyme compared to the free enzyme. Four plasmids, pCEL 16, pCEL 24, pDLG 97 and pDLG 98, were used in this study. BGLA that was cloned into the plasmids pCEL 24 and pDLG 97 was linked to CWP2, and in pDLG 98 it was linked to AGa1 anchor domains. All the plasmids were genome-integrated and expressed in the reference strain Saccharomyces cerevisiae 303-1A. All the transformants were grown in 2% cellobiose and showed higher biomass production compared to the reference strain. β-Glucosidase activity was also assayed and transformed strain W16 showed a fourfold increase in activity compared to the reference strain. There was no significant increase in the activity of the other transformed strains, W24, W97 and W98. Enzymatic characterisation for optimum pH and temperature was done – for all strains the optimum pH was 4 and the optimum temperature was 40ºC. The recombinant strains together with the reference strain were used to make wine from Gewürztraminer grapes. The levels of numerous monoterpenes were enhanced in the resultant wines. The concentration of nerol was increased fourfold, that of citronellol twofold, and geraniol was 20% higher than in the wild type. There was also an increase in the levels of linalool and a-terpinol, but this was not significant. In wines produced with W97, W98 and W24, monoterpene levels did not show a significant difference. In future, the expression of the W16 expression cassette in an industrial wine yeast strain could be performed. In combination with the production of enzymes such as a-arabinofuranosidase, a-rhamnosidase and β-apiosidase, which are involved in the first step of enzymatic hydrolysis, this wine strain could release the bound monoterpenes and enhance the aroma of the wine. / AFRIKAANSE OPSOMMING: Wyn is ‘n komplekse medium. Wynaroma, -geur en -kleur is belangrike kwaliteitsfaktore, hoewel hierdie kwaliteite deur verskeie faktore beïnvloed kan word, soos druifafgeleide verbindings wat as vry vlugtige stowwe teenwoordig kan wees of glikosidies gebind is. Die chemiese samestelling van wyn word bepaal deur faktore soos druifvariëteit, geografiese ligging, wingerdkundige toestande, mikrobiese ekologie van die druif en die wynbereidingsproses. Die variëteitsaroma word bepaal deur vlugtige en nie-vlugtige verbindings, soos monoterpene, norisoprenoïede en benseenderivate, wat natuurlik in die wyn voorkom. Monoterpene is baie belangrik vir die geur en aroma van druiwe en wyn. Monoterpene is teenwoordig in die druiwe en wyn in vry, vlugtige en geurige, of in glikosidiesgebinde, nie-vlugtige en nie-geurige vorms. Die verhouding van glikosidiesgebonde verbindings tot vry aromaverbindings is baie hoog, veral in die Gewürztraminer-, Muscat- en Riesling-kultivars. Glikosidiese verbindings kan deur óf die suurmetode óf die ensiemmetode gehidroliseer word. Die nadeel van die suurmetode is dat dit monoterpene kan modifiseer, terwyl die ensiemmetode die voordeel het dat dit nie die aromakarakter modifiseer nie. Die ensiemmetode waarmee die glikosidiese verbinding afgebreek word, vind in twee opeenvolgende stappe plaas: aanvanklike skeiding van glukose van die terminale suiker deur ‘n hidrolase (a-L-arabinofuranosidase, a-Lramnosidase of β-apiosidase, afhangende van die aglikoongedeelte), gevolg deur die verbreking van die verbinding tussen die aglikoon en glukose deur β- glukosidase. Die β-glukosidase-ensiem kan vanaf ‘n verskeidenheid plant- (Vitis vinifera), bakterie-, gis- en swambronne verkry word. Die meerderheid van die ensieme wat deur hierdie bronne geproduseer word, is nie onder die wynbereidingstoestande van lae pH, hoë temperatuur, hoë glukose en hoë etanol funksioneel nie. β- Glukosidase vanaf ‘n swamoorsprong, veral vanaf Aspergillus-spesies, kan egter wynbereidingstoestande verdra. Die idee agter die gebruik van die β-glukosidasegeen afkomstig van die swam Aspergillus kawachii (BGLA), wat aan die selwand en die vry β-glukosidase gekoppel is, was om te bepaal of die aktiwiteit van die ensiem in vergelyking met dié van die vry ensiem verhoog sou word indien die ensiem aan die selwand geanker is. Vier plasmiede, pCEL 16, pCEL 24, pDLG 97 en pDLG 98, is in hierdie studie gebruik. BGLA, wat in die plasmiede pCEL 24 en pDLG 97 gekloneer is, is gekoppel aan CWP2, en in pDLG 98 is dit aan AGa1-ankergebiede gekoppel. Al die plasmiede is in verwysingsras Saccharomyces cerevisiae 303-1A genoomgeïntegreer en uitgedruk. Al die transformante is in 2% sellobiose gegroei en het hoër biomassaproduksie as die verwysingsras getoon. β-Glukosidaseaktiwiteit is ook geëssaieer en die getransformeerde ras W16 het ‘n viervoudige verhoging in aktiwiteit in vergelyking met die verwysingsras getoon. Daar was geen noemenswaardige verhoging in die aktiwiteit van die ander getransformeerde rasse, W24, W97 en W98, nie. Ensimatiese karakterisering vir optimum-pH en - temperatuur is gedoen – vir al die rasse was die optimum-pH 4 en die optimumtemperatuur 40ºC. Die rekombinante rasse, tesame met die verwysingsras, is gebruik om wyn met Gewürtztraminer-druiwe te maak. Die vlakke van talryke monoterpene is in die gevolglike wyne verhoog. Die konsentrasie van nerol is viervoudig verhoog, dié van sitronellol tweevoudig, en geraniol was 20% hoër as in die wilde tipe. Daar was ook ‘n verhoging in die vlakke van linaloöl en a-terpinol, maar hierdie verhoging was nie noemenswaardig nie. In wyne wat met W97, W98 en W24 gemaak is, het die monoterpeenvlakke nie ‘n noemenswaardige verskil getoon nie. In die toekoms sal die uitdrukking van die W16-uitdrukkingskasset in ‘n industriële wyngisras uitgevoer kan word. In kombinasie met die produksie van ensieme soos a-arabinofuranosidase, a-ramnosidase, β-apiosidase, wat in die eerste stap van ensimatiese hidrolise betrokke is, sal hierdie wyngisras die gebonde monoterpene kan vrylaat en die aroma van die wyn kan verbeter.

Thesis (MSc)--University of Stellenbosch, 2002. / ENGLISH ABSTRACT: No single chemical constituent can be accredited with giving wine and brandy their overall aroma and flavour. The aroma and flavour of wine and brandy are rather attributed to a number of chemical constituents reacting together and it is these reactions that give the beverage its character. Certain chemicals within wine and brandy do, however, make larger contributions to the flavour. These include the esters, terpenes and volatile acids, although others also exist. Esters are a large group of volatile compounds with variable aroma and flavour characteristics, including banana-like (isoamyl acetate), apple-like (ethyl caproate) and chemical/solvent-like (ethyl acetate). Esters are produced as secondary metabolites during the conversion of sugar to ethanol and are formed when an alcohol binds with a fatty acid. Chemically, ester metabolism is well documented and understood; however, much work still needs to be done on a genetic level. The yeast strain used during fermentation is one of the most important factors contributing to the type and quantity of esters produced. This is due to differences in genetic makeup. The metabolism of esters is controlled largely on a genetic level, with numerous genes being involved. The alcohol acetyltransferase genes are involved in ester anabolism, whilst esterase genes are involved in ester catabolism. Esterases have a negative effect on the overall level of esters within an alcoholic beverage, as they are capable of reducing the number of esters and are thus capable of altering the beverage's aroma and flavour profile. The IAH1 and the TIP1 gene products are believed to encode for two such esterases. The objective of this study was to investigate the contribution of the IAH1 and TIP1 genes to the level of esters in both wine and brandy. This was accomplished by using two approaches. Firstly, the above genes were disrupted using a polymerise chain reaction (PCR)-generated disruption cassette homologous to either the IAH1 or the TIP1 gene. These cassettes were integrated into the industrial wine yeast, Saccharomyces cerevisiae strain VIN13. The integrations were verified by Southern blot analysis to produce yeasts VIN13-~IAH1 and VIN13-~TIP1; however, only a single copy of each was disrupted. Secondly, the IAH1 and the TIP1 genes were cloned from S. cerevisiae using PCR into plasmid pj between the phosphoglycerate kinase gene (PGK1) promoter and terminator, producing plasmids pJ-IOE1 and pJ-TOE1. The PGK1 promoter has previously been shown to constitutively express genes at high levels. These new constructs were then used as template for PCR to produce two overexpression cassettes, one for IAH1 and the other for TlP1. These cassettes were integrated into S. cerevisiae VIN13 and verified by Southern blot analysis to produce strains VIN13-IOE1 and VIN13-TOE1. The above yeast strains including VIN13 were used for the production of wines and base wines from Colombard must. Reverse-transcriptase (RT-PCR) confirmed that the VIN13-IOE1 and VIN13-TOE1 strains overexpressed the appropriate gene at a higher level than the control VIN13 strain. The VIN13-AIAH1 disrupted strain showed no difference in expression level to that of the control strain, whilst VIN13-ATIP1 showed lower levels of expression than that of the control strain. VIN13-IOE1 behaved as expected, with a decrease of between 30% and 60% in the total ester level in the wine and base wine respectively, a 30% decrease in the total acid level and no change in the higher alcohol level. The VIN13-AIAH1 strain showed no difference to the control wine, most likely as this strain still expressed the IAH1 gene at levels consistent with the control strain. VIN13-TOE1 behaved in an unexpected manner - instead of hydrolysing esters, it appeared to produce them. This increase in the total ester level was most noticeable during distillation, when a 20% increase took place. Another unexpected occurrence was a large decline in the total acid level, with acetic acid being the most significant contributor, decreasing by up to 78%. This is a very favourable finding, as acetic acid is a known spoilage molecule and is a cause of sluggish/stuck fermentations. VIN13-ATIP1 behaved in an opposite manner to VIN13-TOE1, with higher total acid levels and slightly decreased total ester levels, especially during distillation. Neither affected the total higher alcohol levels. Sensorially, the only significant difference in the wine samples was for the fruity flavour. A panel of judges distinguished that VIN13-TOE1 was fruitier than the other wines, with VIN13-ATIP1 being the least fruity. This study again proves the significant impact that a single gene can have on the chemical makeup of wine and brandy. The relatively simple genetic alteration of an organism can drastically change and improve not only the organoleptic properties of the organism, but its viability as well. These alterations can produce more favourable organisms with more desirable characteristics for the fermenting beverage industry to produce products of higher quality and better suitability. / AFRIKAANSE OPSOMMING: Geen chemiese komponent kan uitgesonderword as die produseerder van aroma en geur in wyn of brandewyn nie. Die aroma en geur van wyn en brandewyn word eerder toegeskryf aan die interaksie tussen 'n groot aantal chemiese komponente om aan die drank sy karakter te gee. Enkele van hierdie chemiese komponente sluit in esters, terpene en vlugtige sure, om maar 'n paar te noem. Esters is "n groot groep van vlugtige verbindings wat beskik oor 'n verskeidenheid van aroma- en geurkenmerke, soos piesangagtig (isoamielasetaat), appelagtig (etielkaproaat) en chemies/oplosmiddelagtig (etielasetaat). Esters word as sekondêre metaboliete geproduseer wanneer suikers na etanolomgeskakel word en word gevorm wanneer "n alkohol met "n vetsuur verbind. Estermetabolisme is chemies goed beskryf en verstaan, maar op "n genetiese vlak is daar nog heelwat aspekte wat nagevors moet word. Die gisras betrokke gedurende fermentasie word beskou as een van die grootse bydraes tot die tipe en die hoeveelheid esters wat geproduseer word. Dit word toegeskryf aan verskille in die genetiese saamestelling van die gisras. Ester metabolisme word grootliks deur genetiese faktore beheer en verskeie gene is betrokke. Dit is hoofsaaklik die alkoholasetieltransferasegene wat vir esterkatabolisme verantwoordelik is, terwyl die esterasegene vir esteranabolisme verantwoordelik is. Esterases het 'n negatiewe effek op die totale estervlak binne alkoholiese dranke deurdat hulle in staat is om die aantal esters drasties te verminder en sodoende die drank se aroma- en geurprofiel te verander. Daar is voorgestel dat die IAH1- en die TlP1-geen produkte is wat vir twee sulke esterases kodeer. Die doel van hierdie studie was om die IAH1- en die TIP1-gene se bydrae tot die totale estervlak in wyn en brandewyn te ondersoek. Dit is deur twee benaderings uitgevoer. Eerstens is die bogenoemde gene d.m.V. disrupsiekassette wat homoloog aan die IAH1- of die TlP1-gene was, uitgeslaan. Die disrupsiekassette is deur die polimerasekettingreaksie (PKR) geproduseer. Hierdie kassette is in die industriële wyngis, Saccharomyces cerevisiae VIN13, geïntegreer. Die integrasies is deur Southernkladanalise bevestig en het die giste VIN13-~IAH1 en VIN13-~TIP1 gelewer. Net 'n enkele kopie van elke geen is egter uitgeslaan. Tweedens is die IAH1- en TIP1-gene d.m.V. PKR vanaf S. cerevisiae binne in plasmied pJ gekloneer, tussen die fosfogliseraatkinasegeen (PGK1) se promotor en termineerder, om plasmiede pJ-IOE1 en pJ-TOE1 te produseer. Die PGK1-promotor is al tevore geïdentifiseer as "n hoë-vlak konstitutiewe uitdrukker van gene. Hierdie twee nuwe konstrukte het vervolgens gedien as templaat vir PKR om twee ooruitdrukkingskassette, een vir IAH1 en die ander vir TIP1, te produseer. Hierdie kassette is in S. cerevisiae VIN13 geïntegreer en bevestig deur Southernkladanalise. Hierdie integrasies het die giste VIN13-IOE1 en VIN13-TOE1 geproduseer. All die nuwe gisrasse, tesame met VIN13, is gebruik vir die produksie van wyne sowel as rebatwyne vanaf Colombard-mos. Omgekeerde-transkriptase polimerasekettingreaksie (OT-PKR) het bewys dat die VIN13-IOE1 en VIN13-TOE1 rasse die geskikte geen ooruitgedruk het, met hoêr vlakke as van die kontrole VIN13-ras. Dit het ook aangedui dat die VIN13-i\IAH1-ras, waarvan die geen uitgeslaan was, geen verskil in uitdrukking gehad het in vergelyking met die kontroleras nie, terwyl VIN13-i\TIP1 'n lae uitdrukkingsvlak getoon het. VIN13-IOE1 het teen verwagting opgetree, met 'n afname van tussen 30% en 60% in die totale estervlak in beide die wyne en rebatwyne. 'n Afname van 30% in die totale suurvlak, asook geen waarneembare verskil in die hoêr alkoholvlak, in vergelyking met die kontroleras, is ook opgemerk. Die VIN13-i\IAH1-ras het glad nie van die kontroleras verskil nie, heel waarskynlik omdat hierdie ras die IAH1-geen teen dieselfde vlak as die kontroleras kon uitdruk. Die VIN13-TOE1-ras het teen verwagting opgetree deurdat dit esters geproduseer het i.p.v. om esters te hidroliseer. Hierdie toename in die totale estervlak is die meeste waarneembaar tydens distillasie, met tot 'n 20% toename. Nog 'n onverwagte effek was die groot afname in die totale suurvlak. met asynsuur wat die betekenisvolste bydrae gelewer het deurdat dit 'n afname van tot 78% getoon het. Hierdie bevinding is baie voordelig, aangesien asynsuur, 'n bekende bederfmolekuul, veral vir slepende/gestaakte fermentasies verantwoordelik is. VIN13-i\TIP1 het op die teenoorgestelde wyse opgetree as VIN13-TOE1, met 'n hoêr totale suurvlak en 'n klein afname in die totale estervlak. Weereens is dit meer gedurende distillasie waargeneem. Beide rasse het egter geen effek op die hoêr alkoholvlak gehad nie. Die proepaneel het, met betrekking tot die vrugtige geur, een betekenisvolle geurverskil tussen die wyne gevind. VIN13-TOE1 was meer vrugtig as al die ander wyne en VIN13-i\TIP1 was die minste vrugtig. Die studie het weereens bewys dat 'n enkele geen 'n betekenisvolle effek op die chemiese samestelling van wyn en brandewyn kan hê. Die relatief eenvoudige genetiese verandering van 'n organisme kan die organoleptiese eienskappe asook die lewensvatbaarheid van "n organisme, drasties verander en verbeter.

O café solúvel aglomerado é produto da aglomeração do café solúvel em pó. A sua produção envolve etapas de processo cujas variações térmicas modificam qualitativamente e quantitativamente a fração de compostos voláteis responsáveis pelo aroma e sabor do produto acabado. A indústria de bebidas de café considera o sabor e aroma de cereal como um defeito na produção de café solúvel aglomerado. Desta forma, este trabalho objetivou aplicar o planejamento experimental para avaliação dos parâmetros de processo da produção de café solúvel aglomerado em escala industrial. Os efeitos destes parâmetros nos componentes voláteis do café e na intensidade do gosto de cereal foram avaliados através do planejamento fracionário 24-1 com resolução IV. As variáveis independentes estudadas foram temperatura do queimador (TQ), fluxo de ar de processo (FA), temperatura da seção quente do leito (TL) e pressão de vapor (PV). Os resultados do planejamento experimental deram-se pela utilização de análise sensorial conduzida por equipe treinada para a detecção do aroma e sabor indesejado. Adicionalmente, os compostos voláteis foram extraídos pela técnica microextração em fase sólida com amostragem no headspace (HS-MFES), identificados e quantificados por cromatografia gasosa acoplada a espectrometria de massa (CG-EM). O efeito dos fatores estudados foi avaliado mediante análise de variância (ANOVA) com nível de significância de 90% e os resultados expressos por gráficos de pareto. A análise de componentes principais (ACP) foi realizada com o intuito de identificar dentre os compostos que podem conferir sabor e aroma de cereais aqueles tem a maior contribuição e quais as condições de processo ideias para evitar a formação do sabor e aroma de cereal. De acordo com ACP foi constatado que piridina, 2-metoxi-4-vinilfenol, 2,3-dimetilpirazina, 2,5-dimetilpirazina, 2,6-dimetilpirazina, 4-metiltiazol, pirazina, 2-metoxifenol, 4,5-dimetiltiazol contribuem para formação do sabor e aroma de cereal. Também indicou que a aglomeração deve ocorrer conforme TQ=215ºC, FA=250rpm, TL=120ºC e PV=1,2 x100KPa. / The agglomerated instant coffee is the product of the powder instant coffee agglomeration. Its production involves stages of processes whose thermal variations change qualitatively and quantitatively the volatile compounds fractions responsible for the aroma and flavor of the finished product. The coffee beverage industry considers the cereal like flavor and aroma as a fault in the production of the agglomerated instant coffee. In this way, this study had as the aim to apply experimental plan to evaluate the parameters of the processes of the agglomerated instant coffee production on an industrial scale. The effects of those parameters in the coffee volatile components and in the intensity of the cereal taste were evaluated through the fractional 24-1 with resolution IV. The independent variables studied were the burner temperature (BT), the process air flow (AF), the temperature of the fluidized bed hot section (LT) and the steam pressure (SP). The results of the experimental plan were made by the use of sensory analysis performed by a trained team for the detection of the undesired aroma and flavor. Additionally, the volatile compounds were extracted by solid-phase microextractiontechniquewithsampling in theheadspace (HS-SPME), identified and quantified by gas chromatography coupled to mass spectrometry (GC-MS). The effect of the factors studied was evaluated by variance analysis (ANOVA) with significance level of 90% and the results showed by Pareto charts. The principal components analysis (PCA) was made with the aim to identify, among several compounds, those that implies on more cereal flavor and aroma, those which major contribution and which ideal process conditions could be used to reduce the cereal flavor and aroma development. According to PCA, it was found that pyridine, 2-metoxi-4-vinylphenol, 2,3-dimethylpyrazine, 2,5-dimetylpyrazine, 2,6-dimethylpyrazine, 4-methyltiazol, pyrazine, 2-methoxyphenol, 4,5-dimethylthiazol contribute to the development of the cereal flavor and aroma. It was also found that the agglomeration must be according to TQ = 215°C, AF=250 rpm, LT=120°C and EPV = 1,2 x 100 KPa.

Le modèle standard supersymétrique minimal (MSSM) est probablement une des théories de nouvelle physique les plus étudiées. Dans la plupart des études phénoménologiques, on fait l'hypothèse de la violation minimale de la saveur dans laquelle toutes les interactions violant la saveur sont directement liées à la matrice CKM du modèle Standard. Cependant, si la supersymétrie est plongée dans des cadres plus larges tels que les théories dites de Grande Unification, de nouvelles sources de violation de la saveur peuvent apparaitre. Les termes correspondant dans le lagrangien sont directement liés au choix du mécanisme de brisure de la supersymétrie. Or, à l'heure actuelle, aucun consensus n’existe quant au choix exact du mécanisme de brisure de la SUSY. Plusieurs mécanismes ont été évoqués récemment (entre autre, par le co-directeur de la thèse), qui pourraient conduire à des signatures prometteuses au LHC pour différents scénarios et domaines de paramètre. Le projet est donc idéalement situé à la frontière entre le 'model building' et des aspects plus phénoménologiques. Pour cette raison, le thésard sera naturellement amené à interagir de manière active avec le groupe de phénoménologie de physique des particules du LAPTh ainsi qu'avec les groupes de physique des particules du LAPP (ATLAS, LHCb..), laboratoire expérimental partageant le même bâtiment que le LAPTh. De plus, des collaborations sur le plan national, en particulier avec le LPSC de Grenoble, ainsi que sur le plan international, avec les universités de Würzburg et Münster (Allemagne) et le nouvel institut de physique de NATAL (Brésil) sont également envisagées. / The Minimal Supersymmetric Standard Model (MSSM) is probably the best studied of all new physics theories. In most phenomenological studies, one assumes the hypothesis of minimal flavour violation, where all flavour-violating interactions are related to the CKM-matrix (as it is the case in the standard model). However, if supersymmetry is embedded in larger frameworks such as grand unified theories (GUT), new sources of flavour violation can appear. The corresponding terms in the Lagrangian are directly related to the mechanism of supersymmetry breaking, for which at present no theoretical consensus exists. Several processes have recently been evoked (among others by the co-director of the proposed thesis), which might lead to promising signatures at the LHC if non-minimal flavour violation is realized. These studies, however, included flavour-violating terms directly at the electroweak scale without taking into account their possible origin at some high scale. The main goal of the project is to extend the previous work in order to take into account the link between the electroweak and the GUT scale. This will allow to put constraints on parameters describing supersymmetry breaking using constraints imposed at the electroweak scale. A second goal is the analysis of relevant processes with the help of current Monte-Carlo simulation tools in order to study the background and thus clarify the observability at LHC for different scenarios and regions of parameters space. So, the project is ideally situated at the frontier between model building and more phenomenological aspects. For this reason the project naturally includes interactions with the local particle physics phenomenology group at LAPTh as well as with the the neighbouring experimental groups (e.g. ATLAS, LHCb) at LAPP. Moreover, collaborations on the national, in particular with the LPSC Grenoble, and international level, e.g. with the University of Würzburg (Germany), the University of Münster (Germany) and the newly created International Institute of Physics in Natal (Brasil) are envisaged.

A esquistossomose é uma doença parasitária aguda e crônica causada por vermes sanguíneos (vermes nematoides) do gênero Schistosoma. O homem contrai a esquistossomose através da penetração ativa da cercaria na pele. A importância do tratamento desta enfermidade consiste não só no fato de curar a doença ou diminuir a carga parasitária dos pacientes, bem como impedir sua evolução para formas mais graves. Para o tratamento da esquistossomose o fármaco de escolha é o praziquantel; isso se deve ao seu amplo espectro, sua eficácia, segurança e a relação custo/tratamento. A única forma farmacêutica disponível no Brasil é o comprimido, na dose de 600 mg, a qual pode ser subdividida em quatro partes de 150 mg, a fim de facilitar o ajuste de dose. No entanto, no momento da subdivisão dos comprimidos ocorre o rompimento do revestimento. Este fato acaba levando a uma exposição do fármaco e, consequentemente, de seu sabor amargo. Esta característica dificulta a administração do medicamento, principalmente na população infantil, prejudicando o tratamento e o controle da doença. Uma alternativa para este problema é o desenvolvimento de sistemas poliméricos microparticulados que associados ao fármaco impediriam o contato direto com as papilas gustativas e assim promoveriam uma melhoria na palatabilidade. Para isso utilizou-se a técnica modificada de deposição interfacial do polímero pré-formado seguido de secagem por aspersão. Três matrizes poliméricas, com diferentes características de liberação foram utilizadas, Eudragit RL 100 – liberação tempo dependente e Eudragit E100 e L30D-55 – liberação pH dependente. Além disso, dois tipos de sistemas carreadores do fármaco foram preparados, microcápsulas e microesferas poliméricas. Estes sistemas obtidos foram avaliados e caracterizados a fim de eleger a melhor proposta de formulação visando o mascaramento do sabor do fármaco. De acordo com os resultados obtidos selecionou-se um sistema composto por microcápsulas formadas a partir do polímero L30D-55. A partir de então inseriu-se este sistema na forma farmacêutica pó para suspensão oral, onde diferentes propostas de formulações, contendo dois edulcorantes auxiliares, aspartame e sacarina, separadamente, e seus respectivos placebos foram avaliadas através de um método in vitro para a determinação do sabor, a língua eletrônica ou sensor gustativo. As diferentes formulações avaliadas apresentaram capacidade em mascarar o sabor desagradável do fármaco e, assim resultam em uma promissora alternativa para o aumento da adesão por parte dos pacientes à terapêutica, principalmente para crianças, em virtude da facilidade de administração, do ajuste da dose em função da massa corpórea e ao sabor muito mais agradável ao paladar infantil. / Schistosomiasis is a parasitic disease acute and chronic caused by blood worms (nematodes worms) of the genus Schistosoma. Man acquires schistosomiasis through the active penetration of the worms in skin. The importance of treatment of this disease is not only the fact of curing the disease or decreases the parasite load of patients, well as prevent progression to more severe forms. For the treatment of schistosomiasis praziquantel is the drug of choice, this is due to its wide spectrum, its efficacy, safety and the relation cost / treatment. The single dosage form available in Brazil is tablet at a dose of 600 mg, which can be subdivided into four parts of 150 mg to facilitate dose adjustment. However when the subdivision of the tablets occurs the disruption of the coating. This fact provides a drug exposure and consequently of its bitter taste. This characteristic complicates the administration of the drug mainly in children, affecting the treatment and control of disease. An alternative for this problem is the development of microparticulate polymeric systems which associated with the drug would prevent direct contact with the taste buds and thus promote an improvement in palatability. For this was used a modified technique interfacial deposition of preformed polymer followed by spray drying. Three polymer matrices with different release characteristics have been used, Eudragit RL 100 – time dependent release, and Eudragit E100 and L30D-55 – pH dependent release. Furthermore, two types of drug carrier systems have been prepared, polymeric microspheres and microcapsules. These systems obtained were evaluated and characterized in order to select the best proposal formulation aimed at masking the taste of the drug. According to the results we selected a system comprising microcapsules formed from L30D-55 polymer. From then was inserted into this system in the pharmaceutical form, powder for oral suspension, where different proposals formulations containing two auxiliary sweeteners, aspartame and saccharin, separately, and their respective placebos were evaluated in an in vitro method for determining the taste, the electronic tongue. The different formulations tested presented excellent ability to mask the unpleasant taste of the drug and thus present an excellent alternative for increasing adherence to therapy, especially for children, because of the ease of administration, according on dose adjustment of body mass and the much more palatable to children's taste.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-02-22T23:54:35Z No. of bitstreams: 1 KarenLuannaMarinhoCatunda_DISSERT.pdf: 1340141 bytes, checksum: 3f6dd7708b6721c9750e0f78ba431a38 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-02-26T21:41:29Z (GMT) No. of bitstreams: 1 KarenLuannaMarinhoCatunda_DISSERT.pdf: 1340141 bytes, checksum: 3f6dd7708b6721c9750e0f78ba431a38 (MD5) / Made available in DSpace on 2016-02-26T21:41:29Z (GMT). No. of bitstreams: 1 KarenLuannaMarinhoCatunda_DISSERT.pdf: 1340141 bytes, checksum: 3f6dd7708b6721c9750e0f78ba431a38 (MD5) Previous issue date: 2015-02-12 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Objetivou-se com esta pesquisa avaliar a influ?ncia do fornecimento de cinco tipos de cact?ceas nativas e ex?ticas do semi?rido do Nordeste brasileiro, sobre as caracter?sticas f?sico-qu?micas, sensoriais e o perfil de ?cidos graxos do leite de cabras da ra?a Saanen. Foram utilizadas cinco cabras mult?paras com nove semanas de lacta??o, peso vivo m?dio inicial de 50 kg ? 4 kg, confinadas e distribu?das em um delineamento quadrado latino 5 x 5, com cinco dietas experimentais e cinco per?odos. Cada per?odo teve dura??o de 17 dias, sendo 10 dias de adapta??o dos animais ?s dietas e sete dias para coleta de dados. Os tratamentos, definidos com base na mat?ria seca, consistiam em: 47,33 a 50,12% de uma cact?cea (xiquexique, mandacaru, facheiro, palma mi?da ou palma orelha de elefante mexicana) mais 18,78 a 19,79% de feno de sabi? e 31,10 a 32,89% de concentrado. N?o foi observado efeito das dietas experimentais na composi??o f?sico-qu?mica do leite para os teores de gordura, s?lidos totais e sais, que apresentaram valores m?dios de 3,24%; 11,30% e 0,66%, respectivamente. No entanto, os teores de prote?na, lactose, extrato seco desengordurado e ponto de crioscopia foram influenciados pelas dietas. No perfil dos ?cidos graxos houve maior concentra??o de ?cidos graxos de cadeias curta e m?dia, contudo, n?o houve altera??es entre os tratamentos, exceto para o ?cido but?rico (C4:0), cujos valores m?dios foram de 4,24% (palma orelha de elefante) a 6,05% (facheiro). As dietas tamb?m n?o conferiram mudan?as sensoriais no leite para os par?metros: odor, flavor, sabor residual e aprecia??o global. A utiliza??o das cinco cact?ceas na alimenta??o de cabras leiteiras n?o influencia as caracter?sticas sensoriais e o perfil lip?dico do leite. A composi??o f?sico-qu?mica do leite se apresenta dentro das exig?ncias m?nimas da legisla??o vigente, exceto para os teores de extrato seco desengordurado e ponto de crioscopia. / The objective of this research was to evaluate the influence of the supply of five types of native and exotic cacti Brazilian semiarid northeast on the sensory characteristics of milk of Saanen goats. Five multiparous goats were used with nine weeks of lactation, average live weight of 50 kg ? 4 kg., confined and distributed in a Latin square design 5 x 5 with five experimental diets and five periods. Each period lasted 17 days, with 10 days of adaptation to the diets of the animals and seven days for the data collection. The treatments were defined based on dry matter consisted of: 47.33 to 50.12% of a cactus (?Xiquexique?, ?Mandacaru?, ?Facheiro? or two species of forage cactus ?Mi?da? or ?Orelha de Elefante Mexicana?) more 18.78 to 19.79% hay of plant ?Sabi?? and 31.10 to 32.89% of concentrate. There was not effect of the experimental diets in the physical and chemical composition of milk for fat, total solids and salts, which showed mean values of 3.24%; 11.30% and 0.66%, respectively. However the protein, lactose, nonfat dry extract and freezing point were affected by diets. In the profile of fatty acids was higher concentration of fatty short and medium chain fatty acids, however, there were not changes between treatments, except for the butyric acid (C4:0), with mean values of 4.24% (?Orelha de Elefante Mexicana?) to 6.05% (?Facheiro?). The diets also did not provide sensory changes in milk for the parameters: odor, flavor, aftertaste and overall assessment. The use of the five cactus in the diet of dairy goats do not influence the sensory characteristics and lipid profile of milk. The physical and chemical composition of milk was showed within the minimum requirements of current legislation, except for nonfat dry extract and freezing point.

A esquistossomose é uma doença parasitária aguda e crônica causada por vermes sanguíneos (vermes nematoides) do gênero Schistosoma. O homem contrai a esquistossomose através da penetração ativa da cercaria na pele. A importância do tratamento desta enfermidade consiste não só no fato de curar a doença ou diminuir a carga parasitária dos pacientes, bem como impedir sua evolução para formas mais graves. Para o tratamento da esquistossomose o fármaco de escolha é o praziquantel; isso se deve ao seu amplo espectro, sua eficácia, segurança e a relação custo/tratamento. A única forma farmacêutica disponível no Brasil é o comprimido, na dose de 600 mg, a qual pode ser subdividida em quatro partes de 150 mg, a fim de facilitar o ajuste de dose. No entanto, no momento da subdivisão dos comprimidos ocorre o rompimento do revestimento. Este fato acaba levando a uma exposição do fármaco e, consequentemente, de seu sabor amargo. Esta característica dificulta a administração do medicamento, principalmente na população infantil, prejudicando o tratamento e o controle da doença. Uma alternativa para este problema é o desenvolvimento de sistemas poliméricos microparticulados que associados ao fármaco impediriam o contato direto com as papilas gustativas e assim promoveriam uma melhoria na palatabilidade. Para isso utilizou-se a técnica modificada de deposição interfacial do polímero pré-formado seguido de secagem por aspersão. Três matrizes poliméricas, com diferentes características de liberação foram utilizadas, Eudragit RL 100 – liberação tempo dependente e Eudragit E100 e L30D-55 – liberação pH dependente. Além disso, dois tipos de sistemas carreadores do fármaco foram preparados, microcápsulas e microesferas poliméricas. Estes sistemas obtidos foram avaliados e caracterizados a fim de eleger a melhor proposta de formulação visando o mascaramento do sabor do fármaco. De acordo com os resultados obtidos selecionou-se um sistema composto por microcápsulas formadas a partir do polímero L30D-55. A partir de então inseriu-se este sistema na forma farmacêutica pó para suspensão oral, onde diferentes propostas de formulações, contendo dois edulcorantes auxiliares, aspartame e sacarina, separadamente, e seus respectivos placebos foram avaliadas através de um método in vitro para a determinação do sabor, a língua eletrônica ou sensor gustativo. As diferentes formulações avaliadas apresentaram capacidade em mascarar o sabor desagradável do fármaco e, assim resultam em uma promissora alternativa para o aumento da adesão por parte dos pacientes à terapêutica, principalmente para crianças, em virtude da facilidade de administração, do ajuste da dose em função da massa corpórea e ao sabor muito mais agradável ao paladar infantil. / Schistosomiasis is a parasitic disease acute and chronic caused by blood worms (nematodes worms) of the genus Schistosoma. Man acquires schistosomiasis through the active penetration of the worms in skin. The importance of treatment of this disease is not only the fact of curing the disease or decreases the parasite load of patients, well as prevent progression to more severe forms. For the treatment of schistosomiasis praziquantel is the drug of choice, this is due to its wide spectrum, its efficacy, safety and the relation cost / treatment. The single dosage form available in Brazil is tablet at a dose of 600 mg, which can be subdivided into four parts of 150 mg to facilitate dose adjustment. However when the subdivision of the tablets occurs the disruption of the coating. This fact provides a drug exposure and consequently of its bitter taste. This characteristic complicates the administration of the drug mainly in children, affecting the treatment and control of disease. An alternative for this problem is the development of microparticulate polymeric systems which associated with the drug would prevent direct contact with the taste buds and thus promote an improvement in palatability. For this was used a modified technique interfacial deposition of preformed polymer followed by spray drying. Three polymer matrices with different release characteristics have been used, Eudragit RL 100 – time dependent release, and Eudragit E100 and L30D-55 – pH dependent release. Furthermore, two types of drug carrier systems have been prepared, polymeric microspheres and microcapsules. These systems obtained were evaluated and characterized in order to select the best proposal formulation aimed at masking the taste of the drug. According to the results we selected a system comprising microcapsules formed from L30D-55 polymer. From then was inserted into this system in the pharmaceutical form, powder for oral suspension, where different proposals formulations containing two auxiliary sweeteners, aspartame and saccharin, separately, and their respective placebos were evaluated in an in vitro method for determining the taste, the electronic tongue. The different formulations tested presented excellent ability to mask the unpleasant taste of the drug and thus present an excellent alternative for increasing adherence to therapy, especially for children, because of the ease of administration, according on dose adjustment of body mass and the much more palatable to children's taste.

Enquanto todos os modelos com neutrinos massivos de Majorana levam ao mesmo operador efetivo de dimensão d = 5, que não conserva número leptônico, os operadores de dimensão d = 6, obtidos a baixas energias, conservam número leptônico e são diferentes dependendo do modelo de alta energia da nova física. Derivamos os operadores de dimensão d = 6 que são característicos de modelos Seesaw genéricos, no qual a massa do neutrino resulta do intercâmbio de campos pesados que podem ser tanto singletos fermiônicos, tripletos fermiônicos ou tripletos escalares. Os operadores resultantes podem conduzir a efeitos observáveis no futuro próximo, se os coeficientes dos operadores de dimensão d = 5 e d = 6 são desacoplados. Neste trabalho apresentamos o modelo violação mínima de sabor no contexto do modelo seesaw tipo III, no qual é possível obter tal desacoplamento. Isto permite reconstruir a estrutura de sabor a partir dos valores das massas dos neutrino leves e dos parâmetros de mistura, mesmo na presença de fases de violação CP. / While all models of Majorana neutrino masses lead to the same dimension five effective operator, which does not conserve lepton number, the dimension six operators induced at low energies conserve lepton number and differ depending on the high energy model of new physics. We derive the low-energy dimension six operators which are characteristic of generic Seesaw models, in which neutrino masses result from the exchange of heavy fields which may be either fermionic singlets, fermionic triplets or scalar triplets. The resulting operators may lead to effects observable in the near future, if the coefficients of the dimension five and six operators are decoupled. In this work we present the model of minimal avor violation in the context of the type III seesaw model, in which it is possible to obtain the decoupling mentioned before. This allows to reconstruct the avour structure of the model from the values of the light neutrino masses and mixing parameters, even in the presence of CP-violating phases.

O típico sabor de chocolate é único, obtido somente de sementes fermentadas, secas e torradas de cacau, não podendo ser sintetizado artificialmente. O desenvolvimento desse sabor é influenciado pela constituição genética das sementes, processamento pós-colheita e manufatura. Proteínas dos cotilédones são potencialmente precursores do sabor e aroma de chocolate. O presente trabalho teve como objetivo analisar diferenças qualitativas e quantitativas nas proteínas de sementes de três genótipos de Theobroma cacao após a colheita e durante a fermentação, de forma a correlacionar estes resultados com diferenças na qualidade (sabor e aroma) obtidas por análise sensorial. Um dos desafios foi o isolamento de proteínas das sementes, evitando o alto teor de polifenóis e polissacarídeos que interferem na separação das proteínas e na análise do proteoma. A metodologia de extração composta por filtração em Miracloth, solubilização e precipitação em ácido ticloroacético (TCA) apresentou géis de maior resolução e repetibilidade, tendo sido escolhida como metodologia de extração protéica para estudo das alterações no proteoma das sementes de cacau durante a fermentação. Foi necessária também a utilização de kit comercial de purificação de proteínas e utilização de método de coloração com nitrato de prata para garantir géis com resolução dos spots e repetibilidade satisfatórias. Os spots foram isolados e após digestão tríptica, submetidos ao sequenciamento por cromatografia líquida associado ao espectrômetro de massas. Os espectros foram analisados pelo programa MASCOT MS/MS Ion Search, utilizando bancos de dados do NCBI. Análises dos mapas 2-D mostraram variação no número de spots entre as variedades. Ao final da fermentação, as proteínas ainda presentes nas sementes das variedades SIAL 70 e Catongo eram ácidas, e o processo de degradação foi caracterizado pelo desaparecimento de quase todas as proteínas neutras ou básicas e também de algumas proteínas ácidas; as proteínas com massa molar acima de 35 kDa também foram todas degradadas. Na variedade CCN 51, não ocorreu o mesmo perfil degradativo, havendo proteínas até pI 6,5 e massa molar acima de 100 kDa. Dos cem spots submetidos ao sequenciamento, 89 foram identificados. As proteínas 21kDa e vicilina foram as proteínas mais abundantes nos cotilédones. Correlacionando os resultados da análise sensorial e a proteômica concluiu-se que existe correlação tanto quantitativa como qualitativa das proteínas dos cotilédones de cacau e possivelmente com as proteínas precursoras de sabor de chocolate / Typical chocolate flavor is unique, only obtained from fermented, dried and roasted cocoa seeds, and can not be synthesized artificially. The flavor development is influenced by the genetic constitution, post-harvest processing and manufactures. Cotyledons proteins are believed to be the precursors of the chocolate flavor. The aim of the present work was to analyze qualitative and quantitative protein differences in seeds of three cocoa genotypes after harvesting and during the fermentation and to correlate these results with differences in quality (flavor and aroma) obtained by sensorial evaluation. One of the challenges was the isolation of proteins from cocoa seeds, avoiding the high content of polyphenols and polysaccharides which disturb protein separation and proteome analysis. The methodology of extraction by filtration in Miracloth, solubilization and precipitation in trichloroacetic acid showed the highest gel resolution and reprodutivity, and, thus, was chosen to be used in the analyses of the proteome of cocoa seeds during the fermentation. It was also necessary to use commercial kit for protein purification and a silver-based staining method with nitrate to guarantee gels with spots resolution and satisfactory reproducibility. Proteins were excised from de gels and after tryptic digestion, MS analysis was conducted by on line chromatografhy using a Cap-LC coupled to a mass spectrometer. The spectra were processed using MASCOT MS/MS Ion Search, and the sequences searched against NCBI databases. The 2-DE maps analysis of cocoa seeds showed significant variation of the spots number among the genotypes. At the end of fermentation, proteins still present in the Sial 70 and Catongo genotypes were acid and the degradation process was characterized by the disappearance of almost all the neutral or basic proteins and also some acid proteins. The genotype CCN 51 did not show the same degradation profile. Of the spots submitted to the mass spectrometer, 89 were identified. The 21kDa protein and vicilin were the most abundant proteins in the cocoa cotyledons. Correlating sensorial analysis and the proteomic results we could observe the existence of quantitative as qualitative correlation of proteins from cocoa cotyledons and possibly with the precursors proteins of chocolate flavor