京都大学 / 新制・課程博士 / 博士(情報学) / 甲第24740号 / 情博第828号 / 新制||情||139(附属図書館) / 京都大学大学院情報学研究科数理工学専攻 / (主査)教授 梅野 健, 教授 山下 信雄, 准教授 加嶋 健司 / 学位規則第4条第1項該当 / Doctor of Informatics / Kyoto University / DFAM

Glucosyltransferases (GTs) are the important group of enzymes which facilitates the incorporation of UDPactivated glucose to a corresponding acceptor molecule through glucosylation. Glucosylation is a common alteration reaction in plant metabolism and is regularly associated with the production of secondary metabolites. Glucosylation serves a number of roles within metabolism including: stabilizing structures, affecting solubility, transport, and regulating the bioavailability of the compounds for other metabolic processes. GTs involved in secondary metabolism share a conserved 44 amino acid residue motif (60–80% identity) known as the plant secondary product glucosyltransferase (PSPG) box, which has been demonstrated to include the UDP-sugar binding moiety. Among the secondary metabolites, flavonoid glycosides affect taste characteristics in citrus making the associated glucosyltransferases particularly interesting targets for biotechnology applications in these species. Custom design of enzymes requires understanding of structure/function of the protein. The present study focuses on creating mutant Flavonol- 3-O- Glucosyltransferases proteins using site-directed mutational analysis and testing the effect of each mutation on substrate specificity and kinetic properties of the enzyme.

Studies of the relationship between market values and accounting numbers have long been a part of an established theme in capital markets research (CMR). These studies have taken various forms, most being conducted on a cross sectional basis, tied closely with the assumptions of equilibrium behaviour and efficient markets. Explanatory variables for market value have been dominated by firm-specific variables without incorporating macroeconomic variables. Recently, however, some studies have employed macroeconomic variables and dynamic specification in assessing the relationship between market values and accounting numbers (e.g. Bilson et al. 2001, Nissim and Penman, 2003, and Willett, 2003). The objective of this thesis is to investigate the nature of the relationship between share prices and accounting numbers on the Jakarta Stock Exchange for the period 1992-2002, using dynamic modelling principles in addition to the more usual cross sectional analysis. The approach to regression modelling (general-to-specific strategy)incorporated in this thesis relies less heavily than most CMR on prior economic theories of equilibrium behaviour. Apart from these novel aspects of approach and method, the study also provides valuable information about the emerging financial markets of Indonesia. The results of this thesis show that cointegration and the accompanying equilibrium correction relationship between market and book values for firms listed on the Jakarta Stock Exchange (JSX) can often be identified using accounting and macroeconomic regressors. The models are typically more informative, plausible and consistent than cross sectional models and are useful in interpreting the context in which the market to book relationship exists in Indonesia. A possibly surprising result is that in Indonesia, compared to similar models estimated using US data, the book value of net assets seems to have a stronger relationship with market value. This may be a function of the relative importance of financial statements as a source of information on the JSX.

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Em próteses sobre implantes, o recobrimento oclusal com materiais pouco rígidos como os acrílicos, é por vezes utilizado por supostamente reduzir a carga transmitida ao osso, entretanto não existe um consenso na literatura sobre a sua efetividade. O objetivo deste trabalho foi comparar, por análise fotoelástica e dinâmica, a transmissão de forças após a ação de cargas sobre próteses unitárias sobre implantes em função dos materiais protéticos. Confeccionou-se um modelo em resina fotoelástica com anatomia semelhante à região de pré-molares inferiores, contendo um implante de hexágono interno de 3,75 x 11,5 mm. Foram elaboradas três tipos de próteses em materiais com rigidez decrescente: 1- Metálica (alta rigidez); 2- Resina composta (rigidez moderada), 3- Com um disco de EVA interposto com h=2,5 mm (baixa rigidez). Por meio de um polariscópio circular, observou-se qualitativamente a tensão gerada no modelo durante a aplicação de forças axiais de 100 N, em regime compressivo, sobre cada tipo de prótese. Para a análise dinâmica, realizou-se um modelamento físico de cada um dos três sistemas implante-prótese com modelos de Voigt. A partir daí, uma representação matemática dos sistemas foi deduzida e resolvida, levando em consideração as propriedades mecânicas dos diferentes materiais testados, como o módulo de elasticidade (E), o coeficiente elástico (k) e a constante de amortecimento (c). Para representar uma carga mastigatória fisiológica, foi aplicado um pulso de força senoidal com duração de 0,25 s e intensidade máxima de 100 N, segundo a expressão ()) 8 cos( . 50 50 t t f π −=. Observou-se na análise fotoelástica que após a aplicação de 100N, surgiram tensões na região apical e na cervical dos implantes, com intensidade e localização similares entre os três tipos de prótese. A análise dinâmica mostrou que a força de 100N aplicada na prótese é totalmente transmitida ao implante instantaneamente. Os resultados das análises concordam entre si e mostram que não é possível amortecer as forças mastigatórias por meio dos materiais oclusais. / Less rigid oclusal materials such as acrylic resins are often used in prosthesis over implants because they are supposed to diminish the loads transferred to the bone; however there is no scientific evidence to support this theory. The objective of this work was to compare by photoelastic and dynamic analysis the load transfer after oclusal loads over single prosthesis over implants, varying the oclusal materials. A photoelastic model was created, with an anatomy similar to an inferior premolar region. An internal hexagon implant (3,75 x 11,5 mm) was put in this model. Three different types of prosthesis were constructed with decrescent stiffness: 1- Metallic (higher stiffness); 2- Composite resin (moderate stiffness); 3- With an EVA disc interposed (fewer stiffness). By means of a circular polariscope, the stress generated in the model with a compressive axial load of 100 N, was observed for each prosthesis. For the dynamic analysis, a physical modeling of each implant-prosthesis was carried out with Voigt models. After that, a mathematical representation of the systems was deduced and solved, taking into account the mechanic properties of the different materials, like the elastic modulus (E), the elastic coefficient (k) and the damping constant (c). To represent a physiologic masticatory load, a senoidal force was applied, during 0,25 s and maximum load of 100 N, by the expression ()) 8 cos( . 50 50 t t f π −=. With the photoelastic analysis, it was observed that stresses were generated at the apical and cervical regions of implants, with similar intensity and patterns between the three types of prosthesis. The dynamic analysis, showed that the loads applied over the prosthesis are transferred to the implant instantaneously. The results of both anaylis are in agreement that it is not possible to damp masticatory loads by means of occlusal materials.

Flavonoids are an important class of secondary metabolites widely distributed in plants. The majority of naturally occurring flavonoids are found in glucosylated form. Glucosyltransferases are enzymes that enable transfer of glucose from an activated donor (UDP-glucose) to the acceptor flavonoid substrates. A flavonol specific glucosyltransferase cloned from Citrus paradisi (Cp3OGT) has strict substrate and regiospecificity. In this study, amino acid residues that could potentially alter the rigidity observed in this enzyme were mutated to position equivalent residues of a putative anthocyanin specific glucosyltransferase from Clitorea ternatea and a GT from Vitis vinifera that can glucosylate both flavonols and anthocyanidins. Using homology modeling followed by site directed mutagenesis to identify candidate regions, three double mutations were made. To test the basis of substrate specificity, biochemical analysis of the three recombinant mutant proteins was carried out. Recombinant protein with mutation S20G+T21S revealed that the enzyme retained activity similar to the wildtype (Cp3OGT) (WT- Km app-104.8 µM; Vmax = 24.6 pmol/min/µg, Mutant- Km app-136.42 µM; Vmax -25pmol/min/µg) but the mutant was more thermostable compared to the WT. The (S290C+S319A) mutant protein retained 40% activity relative to wildtype and has an optimum pH shifted towards the acidic side (pH 6) (Km app-8.27 µM; Vmax-90.9 pmol/min/µg). Mutation of Glutamine87 and Histine154 (H154Y+Q87I) have rendered this recombinant protein inactive with every class of flavonoid tested. Interestingly, the single point mutations H154Y and Q871I had significant activity, slightly greater than that of wildtype enzyme. The two active recombinant proteins will further be analyzed to determine whether the mutations have altered regiospecificity of the original enzyme. Product identification is being conducted using HPLC.

Glucosyltransferases (GTs) are enzymes that expedite the incorporation of UDP-activated glucose to a corresponding acceptor molecule. This enzymatic reaction stabilizes structures and affects solubility, transport, and bioavailability of flavonoids for other metabolic processes. Flavonoid glycosides affect taste characteristics in citrus making the associated glucosyltransferases particularly interesting targets for biotechnology applications. Custom design of enzymes requires understanding of structure/function of the protein. The present study focuses on creating mutant flavonol-3-O-glucosyltransferase (F-3-O-GT) proteins using site-directed mutagenesis and testing the effect of each mutation on substrate specificity, regiospecificity and kinetic properties of the enzyme. Mutations were selected on the basis of sequence similarity between grapefruit F-3-O-GT, an uncharacterized GT gene in blood orange (98%), and grape F3GT (82%). Grapefruit F-3-O-GT prefers flavonol as a substrate whereas the blood orange sequence is annotated to be a flavonoid 3GT and the grape GTs could glucosylate both flavonols and anthocyanidins. Mutants of F-3-O-GT were generated by substituting L41M, N242K, E296K and N242K+E296K and proteins were expressed in Pichia pastoris using the pPICZA vector. Analysis of these mF-3-O-GTs showed that all of them preferred flavonols over flavanone, flavone, isoflavones, or anthocyanidin substrates and showed decrease in enzyme activity of 16 to 51% relative to the wild type F-3-O-GT.

Differential Item Functioning Analyses investigates whether individuals with same ability in different groups also show similar performance on an item. In matching the individuals of the same ability, most of the methodologies use total scores of the tests which are usually constructed to be unidimensional. th purpose of the present study is evaluating the PISA 2003 mathematics literacy items through the use of DIF methodology which uses a multidimensional approach in matching students instead of single total score, improve the matching for DIF analyses. In the study factor structure of the tests will be determeined via both exploratory and confirmatory analyses in a complimentary fashion. then DIF analyses conducted using Logistic regression (LR) and Mantel -Haenszel methods.Analyses showed that the matching criterion improved when multivariate analyses were used. the number of DIF items was decreased when the matching criterion is defined based on multiple criterion scores such as mathematical literacy and problem solving scores or two different mathematical literacy subtest score. In addition, qualitative reviews and examination of the distribution of DIF items by content categories, cognitive demands, item types,item text, visual-spatial factors and linguistic properties of items were analyzed to explain the differential performance. Curriculum, cultural and translation differences were the main criteria for the qualitative analyses of DIF items. The results imply that curriculum and translation differences in items might be causing the DIF across Turkish and English versions of the tests.

Em próteses sobre implantes, o recobrimento oclusal com materiais pouco rígidos como os acrílicos, é por vezes utilizado por supostamente reduzir a carga transmitida ao osso, entretanto não existe um consenso na literatura sobre a sua efetividade. O objetivo deste trabalho foi comparar, por análise fotoelástica e dinâmica, a transmissão de forças após a ação de cargas sobre próteses unitárias sobre implantes em função dos materiais protéticos. Confeccionou-se um modelo em resina fotoelástica com anatomia semelhante à região de pré-molares inferiores, contendo um implante de hexágono interno de 3,75 x 11,5 mm. Foram elaboradas três tipos de próteses em materiais com rigidez decrescente: 1- Metálica (alta rigidez); 2- Resina composta (rigidez moderada), 3- Com um disco de EVA interposto com h=2,5 mm (baixa rigidez). Por meio de um polariscópio circular, observou-se qualitativamente a tensão gerada no modelo durante a aplicação de forças axiais de 100 N, em regime compressivo, sobre cada tipo de prótese. Para a análise dinâmica, realizou-se um modelamento físico de cada um dos três sistemas implante-prótese com modelos de Voigt. A partir daí, uma representação matemática dos sistemas foi deduzida e resolvida, levando em consideração as propriedades mecânicas dos diferentes materiais testados, como o módulo de elasticidade (E), o coeficiente elástico (k) e a constante de amortecimento (c). Para representar uma carga mastigatória fisiológica, foi aplicado um pulso de força senoidal com duração de 0,25 s e intensidade máxima de 100 N, segundo a expressão ()) 8 cos( . 50 50 t t f π −=. Observou-se na análise fotoelástica que após a aplicação de 100N, surgiram tensões na região apical e na cervical dos implantes, com intensidade e localização similares entre os três tipos de prótese. A análise dinâmica mostrou que a força de 100N aplicada na prótese é totalmente transmitida ao implante instantaneamente. Os resultados das análises concordam entre si e mostram que não é possível amortecer as forças mastigatórias por meio dos materiais oclusais. / Less rigid oclusal materials such as acrylic resins are often used in prosthesis over implants because they are supposed to diminish the loads transferred to the bone; however there is no scientific evidence to support this theory. The objective of this work was to compare by photoelastic and dynamic analysis the load transfer after oclusal loads over single prosthesis over implants, varying the oclusal materials. A photoelastic model was created, with an anatomy similar to an inferior premolar region. An internal hexagon implant (3,75 x 11,5 mm) was put in this model. Three different types of prosthesis were constructed with decrescent stiffness: 1- Metallic (higher stiffness); 2- Composite resin (moderate stiffness); 3- With an EVA disc interposed (fewer stiffness). By means of a circular polariscope, the stress generated in the model with a compressive axial load of 100 N, was observed for each prosthesis. For the dynamic analysis, a physical modeling of each implant-prosthesis was carried out with Voigt models. After that, a mathematical representation of the systems was deduced and solved, taking into account the mechanic properties of the different materials, like the elastic modulus (E), the elastic coefficient (k) and the damping constant (c). To represent a physiologic masticatory load, a senoidal force was applied, during 0,25 s and maximum load of 100 N, by the expression ()) 8 cos( . 50 50 t t f π −=. With the photoelastic analysis, it was observed that stresses were generated at the apical and cervical regions of implants, with similar intensity and patterns between the three types of prosthesis. The dynamic analysis, showed that the loads applied over the prosthesis are transferred to the implant instantaneously. The results of both anaylis are in agreement that it is not possible to damp masticatory loads by means of occlusal materials.

Flavonoids are secondary metabolites that are important in plant defense, protection and human health. Most naturally-occurring flavonoids are found in glucosylated form. Glucosyltransferases (GTs) are enzymes that catalyze the transfer of glucose from a high energy sugar donor to an acceptor molecule. A flavonol-specific 3-O-GT enzyme has been identified and cloned from leaf tissues of grapefruit. The enzyme shows rigid substrate specificity and regiospecificity. F3-O-GTs from grape (Vitis vinifera) and grapefruit (Citrus paradisi) were modeled against F7-O-GTs from Crocus sativus and Scrutellaria biacalensis, and several non-conservative amino acid differences were identified that may impact regioselectivity. This research is designed to test the hypothesis that specific amino acid residues impart the regiospecificity of the grapefruit enzyme. Site-directed mutagenesis was performed on three potentially key amino acid residues within the grapefruit F3-O-GT that were identified through homology modeling. Analyses of the enzyme activity of the mutant F3-O-GT proteins revealed that the single point mutations of serine 20 to leucine (S20L) and proline 297 to phenylalanine (P297F) rendered the recombinant enzyme inactive with flavonol substrates. Mutation of glycine 392 to glutamate (G392E) was active at 80% relative to the wild type. The mutant enzyme also did not show broadened acceptor specificity as it also favored flavonols as the preferred acceptor substrate. The glucosylation products of the active mutant enzyme will be analyzed to determine if this resulted in a change in regiospecificity.