Due to an increased environmental awareness and thus, concerns over the use of fossil-based monomer for polymer production, there is an ongoing effort to find alternatives to non-renewable traditional monomers. This has ushered in the rapid growth in the development of bio-based materials such as green monomers and biodegradable polymers from vegetable and animal resources. Glycerol, as a renewable bio-based monomer, is an interesting candidate for sustainable polymer production. Glycerol is a renewable material that is a by-product of the transesterification of vegetable oils to biodiesel. Utilization of the excess glycerol derived from the growing biodiesel industry is important to oleochemical industries. The main objective of this thesis was to produce high molecular weight polyglycerol from glycerol and synthesize stimuli-responsive polyglycerol hydrogels. The work began with an investigation of the step-growth polymerization of glycerol to relatively high molecular weight polyglycerol using several catalysts. The catalytic reaction mechanisms were compared and the polymer products were fully analyzed. High molecular weight partially branched polyglycerol with multimodal molecular weight distributions was obtained. The polymerization of glycerol proceeded fastest with sulphuric acid as catalyst as indicated by the highest observed conversion of monomer along with the highest molecular weights. Theoretical models were used to predict the gel point and to calculate monomer functionality. High molecular weight polyglycerol was used to synthesize novel stimuli-responsive hydrogels. Real-time monitoring of step-growth polymerization of glycerol was investigated using in-line and off-line Attenuated Total Reflectance/Fourier Transform infrared (ATR-FTIR) technique.

RM257 is a photopolymerisable diacrylate mesogen. In its normal way, the polymerization of the diacrylate mesogen is initiated by using UV light. RM257 mesogen have photopolymeriable acrylate endgroups. RM257 molecules form to polymer network. In this study, we use 632.8 nm HeNe laser to initiate the monomer conversion of RM257. The effects of light intensity, exposure time and temperature of monomer on the rate of polymerization have been studied. The monomer conversion of RM257 is found to have a higher rate when the mesogen in the isotropic phase. On other hand, using a two ¡Vbeen interference pattern, a periodic structure was be produced in the RM257 polymer network. The mesogen has a uniform ordered. To discuss that diacylate effected by holographic light field.

Due to an increased environmental awareness and thus, concerns over the use of fossil-based monomer for polymer production, there is an ongoing effort to find alternatives to non-renewable traditional monomers. This has ushered in the rapid growth in the development of bio-based materials such as green monomers and biodegradable polymers from vegetable and animal resources. Glycerol, as a renewable bio-based monomer, is an interesting candidate for sustainable polymer production. Glycerol is a renewable material that is a by-product of the transesterification of vegetable oils to biodiesel. Utilization of the excess glycerol derived from the growing biodiesel industry is important to oleochemical industries. The main objective of this thesis was to produce high molecular weight polyglycerol from glycerol and synthesize stimuli-responsive polyglycerol hydrogels. The work began with an investigation of the step-growth polymerization of glycerol to relatively high molecular weight polyglycerol using several catalysts. The catalytic reaction mechanisms were compared and the polymer products were fully analyzed. High molecular weight partially branched polyglycerol with multimodal molecular weight distributions was obtained. The polymerization of glycerol proceeded fastest with sulphuric acid as catalyst as indicated by the highest observed conversion of monomer along with the highest molecular weights. Theoretical models were used to predict the gel point and to calculate monomer functionality. High molecular weight polyglycerol was used to synthesize novel stimuli-responsive hydrogels. Real-time monitoring of step-growth polymerization of glycerol was investigated using in-line and off-line Attenuated Total Reflectance/Fourier Transform infrared (ATR-FTIR) technique.

Made available in DSpace on 2014-06-11T19:28:56Z (GMT). No. of bitstreams: 0 Previous issue date: 2003-02-27Bitstream added on 2014-06-13T19:16:53Z : No. of bitstreams: 1 urban_vm_me_arafo.pdf: 1219036 bytes, checksum: 4e3be4eb79ce75a36f6d11fd248eff4f (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O objetivo deste estudo foi estabelecer, inicialmente, um método analítico em CLAE, para comparar o conteúdo de monômero residual de uma resina autopolimerizável para reembasamento imediato (Duraliner II - D) e uma resina termopolimerizável utilizada na confecção de bases de próteses (Lucitone 550 - L). Além disso, foi avaliado o efeito de dois tratamentos térmicos sobre esse conteúdo. Os monômeros utilizados como padrões foram o MMA (resina L) e o BMA (resina D). As condições cromatográficas para a análise foram: coluna ODS (15,0 x 0,46 cm), com 5 mm de tamanho de partícular e 100 Å de tamanho de poro, fase móvel ACN : H2O (55%: 45%) + 0,01% TEA, fluxo 0,8 mL/min, l 230 nm, volume de injeção 100 mL. As resinas foram proporcionadas e polimerizadas de acordo com as instruções dos fabricantes. Os corpos-de-prova (n = 18) dos dois materiais foram divididos em 3 grupos, com 6 corpos-de-prova cada. O grupo I (GI) não foi submetido a nenhum tratamento (controle). Para o grupo II (GII), os corpos-de-prova foram irradiados com microondas, a 550 W por 3 min para o material L e a 650 W por 4 min para o material D. Para o grupo III (GIII), os corpos-de-prova dos materiais L e D foram submetidos ao tratamento térmico por imersão em água a 55oC por 60 min e 10 min, respectivamente. Cada corpo-de-prova foi, então, fragmentado e submetido ao procedimento de extração em MeOH por 2 horas. Os valores de conteúdo de monômero residual foram transformados em porcentagem e analisados pelo teste de Kruskall-Wallis, ao nível de confiança de 99%. Em todos os grupos avaliados, o material D apresentou valores médios de monômero residual (GI - 0,87%; GII - 0,71%; GIII - 0,52%) significativamente maiores (p<0,01) que o material L (GI - 0,09%; GII - 0,06%; GIII - 0,05%). De acordo com a metodologia utilizada neste estudo, foram obtidas as seguintes conclusões: 1) O método... / The aim of this study was to firstly establish an analytical method, which was the most satisfactory with reference to optimization of chromatographic conditions for HPLC. The method was then used to compare the residual monomer content in one hard chair-side reline resin (Duraliner II - D) and one heat-polymerized denture base resin (Lucitone 550 - L). Also, it was investigated the effect of two after polymerization heat-treatments on this residual monomer content. The HPLC operating conditions established were column ODS (15.0 x 0.46 cm), with 5 mm particles, mobile phase - 55% ACN : 45% deionized distilled water, flow injection rate - 0.8 mL/min., wave length detection - UV 230 nm, amount of sample solution injection 100 mL. Calibration curves were produced using standard solutions containing known amounts of each monomer evaluated (MMA - L; BMA - D) in mobile phase. The materials (D and L) were mixed and polymerized according to the manufacturer's instructions. Specimens (n = 18) of both materials were then divided into three groups, of six specimens each. Group I specimens were left untreated (GI - control). Specimens of group II (GII) were given heat treatment by microwave irradiation with 550 W for 3 min for material L and 650 W for 4 min for material D. For group III (GIII), L and D specimens were submitted to a heat-treatment by immersion in water at 55oC for 60 min and 10 min, respectively. Each specimen were cut into small pieces and subjected to an extraction procedure in methanol for 2 hours. The residual monomer content was then expressed as a percentage of residual monomer in the specimen. The percentage residual monomer data were analyzed by non-parametric Kruskall-Wallis test, at 99% level of confidence. In all groups evaluated, material D showed significantly higher (p<0.01) residual monomer mean values (GI - 0.87%; GII - 0.71%; ...(Complete abstract, click electronic access below)

Acanthamoeba mannose-binding protein (AcMBP) is a virulence factor of the free-living amoeba, Acanthamoeba castellanii. It is crucial for the development of Acanthamoeba keratitis (AK), a corneal infection that often causes blindness. AK is associated with contact lens use and contaminated water sources. Therapeutic unresponsiveness is attributed to similarities in the biological processes that Acanthamoeba shares with humans and its ability to form drug-resistant cysts. I aimed to characterise AcMBP as a basis for developing future drugs against Acanthamoeba. To start with, I carried out morphological studies on the two well-known life stages of Acanthamoeba and characterised a third stage: the protocysts. Mature cysts and protocysts could not interconvert directly, but always excysted to trophozoites. This is important because Acanthamoeba can potentially be trapped as protocysts, which are likely to be more susceptible to drugs. I also studied Acanthamoeba adhesion towards various surfaces and cytopathic activities towards cells (including human corneal epithelial cells). Whilst AcMBP was important for adhesion, it is not the only receptor involved. To gain structure/function information, I expressed the extracellular portion of AcMBP and three truncated fragments. AcMBP is a Ca2+-dependent lectin (~100 kDa) that binds to mannose. Ca2+ is essential for lectin activity and stability. The extracellular fragment is monomeric, indicating that trimerisation, shown previously, depends on the membrane-spanning and/or intracellular regions. Bioinformatics revealed that lectin activity is almost certainly located in a DUF 4114 domain (~10 kDa, DUF: domain of unknown function). N-terminal fragments, including the DUF4114 domain did not bind to mannose-Sepharose, suggesting that part of the cysteine-rich domain is also important. AcMBP bound to a variety of mammalian glycans so may have more than one lectin activity. Although attempts to crystallise AcMBP were unsuccessful, future structural analysis will be useful for defining the domains and determining how it binds to mannose.

There has been much interest in recent years about the potential of microRNA as a new source of biomarkers for the diagnosis of disease. The delivery of new diagnostic tools based on this potential has been limited by shortcomings in current microRNA detection techniques. This thesis explores the development of a new method of microRNA detection through the incorporation of conductive particles into oligonucleotide-functionalised polymers to form oligonucleotide cross-linked polymer composites. Such composites could provide a simple, rapid, and low-cost means of microRNA detection that could be easily multiplexed, providing a valuable tool for point-of-care medical diagnostics. This work presents oligonucleotide-functionalised carbon/polyacrylamide composites which demonstrate a selective swelling response in the presence of analyte oligonucleotide sequences and for which the electrical conductivity decreases with swelling. The composites were synthesised via UV-initiated free-radical polymerisation of carbon/- monomer mixtures upon custom electrode devices, consisting of interdigitated platinum electrodes fabricated upon a silicon substrate. The optimal cross-linker density and carbon loading concentration were determined as well as the best means of dispersing the carbon particles within the polymer. Various types of carbon particles, with differing sizes and aspect ratios, were compared and their performances as conductive additives for polymer swelling transduction evaluated. The swelling behaviour of these composites was evaluated by analysing images of composite microdroplets as they swell. The electrical characteristics of the composites were determined by measuring either the two-terminal resistance or the complex impedance of composite microdroplets on the electrode devices. Alternating and direct current measurement techniques were compared to determine the best approach for the transduction of composite swelling. The volumetric and electrical responses of oligonucleotide-functionalised carbon/polyacrylamide composites were analysed in solutions of analyte oligonucleotide and non-complementary controls. It has been demonstrated that, using carbon nanopowder composites and a direct current two-terminal resistance measurement, it is possible to differentiate between analyte and control solutions to concentrations as low as 10 nM, with single-base precision, in less than three minutes. However, the inability to detect at concentrations below this value, difficulties in differentiating between different analyte concentrations and thermal instability mean that, in their current form, oligonucleotide cross-linked polymer composites are unsuitable for the detection of circulating microRNA at clinically relevant concentrations. Potential avenues of work to address these challenges are discussed. Also presented are collaborative results for oligonucleotide-responsive polymers functionalised with morpholino nucleic acid analogues, in what is believed to be the first example of such a material. These morpholino-functionalised polymers offer significant advantages, in terms of stability and sensitivity, over their nucleic acid equivalents for bio-responsive polymer applications.

A hemoglobina (Hb) do anelídeo Glossoscolex paulistus é uma Hb extracelular gigante. Esta é constituída por quatro cadeias polipeptídicas a, b, c e d contendo o heme. As cadeias a, b e c formam um trímero mantido por ligações dissulfeto e a cadeia d é um monômero. Neste trabalho, as técnicas espectroscópicas de absorção ótica no UV-Vís, fluorescência e dicroísmo circular foram utilizadas para monitorar as mudanças produzidas na estrutura do monômero d na forma oxi da hemoglobina extracelular gigante de Glossoscolex paulistus (HbGp) em pH 7,0 e 9,0 devido à interação com surfactantes iônicos, dodecil sulfato de sódio (SDS), cloreto de cetiltrimetilamônio (CTAC) e neutro, polioxietileno lauril éter (BRIJ - 35). A interação entre surfactantes e proteínas globulares tem sido estudada através de uma variedade de técnicas espectroscópicas visando à obtenção de informações relevantes com respeito à relação estrutura-atividade das proteínas. Os resultados de fluorescência mostraram que na presença dos surfactantes, há um aumento do rendimento quântico, sendo que o maior aumento é verificado na interação com o surfactante catiônico CTAC, sugerindo que a proteína apresenta uma quantidade maior de sítios específicos carregados negativamente, como os resíduos aniônicos, glutamil e aspartil. Na interação do monômero d da HbGp com CTAC ocorreu a formação de espécie pentacoordenada em ambos os pHs. Para o sistema monômero da HbGp-SDS em pH 7,0 e 9,0, foi observada a oxidação do monômero d, caracterizada pela formação da espécie hemicromo. A interação do monômero d em pH 7,0 e 9,0 com CTAC, analisada pelos espectros de dicroísmo circular, indicaram uma perda não muito significativa da estrutura secundária. No entanto, para os surfactantes SDS e BRIJ-35 foi possível constatar uma perda menos acentuada em relação ao CTAC. / The annelid hemoglobin (Hb) of the Glossoscolex paulistus is a giant extracellular hemoglobin (Hb). It is constituted by four polypeptide a, b, c and d chains containing the heme. The chains a, b and c form a trimer maintained by disulfide bond and the d chain is a monomer. In this work, the spectroscopic techniques of UV-Vís optical absorption, fluorescence and circular dichroism were used to monitor the changes induced by ionic detergents sodium dodecyl sulphate (SDS), cetyltrimethylammonium chloride (CTAC) and neutral, ether lauryl polioxitelene (BRIJ-35) in the structure of the d monomer in the oxi form of the giant extracellular hemoglobin of Glossoscolex paulistus (HbGp) in pH 7.0 and 9.0. The interaction between surfactants and globular proteins has been studied through a range of spectroscopic techniques targeting the obtaining of prominent information with regard to the relation structure-activity of the proteins. The fluorescence results showed that in the presence of the surfactants a quantum yield increases, and the CTAC promoted the more significant effect, suggesting that the protein presents a larger quantity of specific negative sites, as the anionic residues, glutamil and aspartil. In the interaction of d monomer of the HbGp with CTAC occurred the formation of pentacoordinated species in both pHs. For system monomer of HbGp-SDS at pH 7.0 and 9.0, was observed by oxidation of d monomer, characterized by the formation of the species hemichrome. The interaction of d monomer at pH 7.0 and 9.0 with CTAC, analyzed by circular dichroism spectra, indicates the loss of secondary structure. However, for SDS and BRIJ-35 surfactants it is possible to detect the lower loss of secondary structure in relation to CTAC.

Due to an increased environmental awareness and thus, concerns over the use of fossil-based monomer for polymer production, there is an ongoing effort to find alternatives to non-renewable traditional monomers. This has ushered in the rapid growth in the development of bio-based materials such as green monomers and biodegradable polymers from vegetable and animal resources. Glycerol, as a renewable bio-based monomer, is an interesting candidate for sustainable polymer production. Glycerol is a renewable material that is a by-product of the transesterification of vegetable oils to biodiesel. Utilization of the excess glycerol derived from the growing biodiesel industry is important to oleochemical industries. The main objective of this thesis was to produce high molecular weight polyglycerol from glycerol and synthesize stimuli-responsive polyglycerol hydrogels. The work began with an investigation of the step-growth polymerization of glycerol to relatively high molecular weight polyglycerol using several catalysts. The catalytic reaction mechanisms were compared and the polymer products were fully analyzed. High molecular weight partially branched polyglycerol with multimodal molecular weight distributions was obtained. The polymerization of glycerol proceeded fastest with sulphuric acid as catalyst as indicated by the highest observed conversion of monomer along with the highest molecular weights. Theoretical models were used to predict the gel point and to calculate monomer functionality. High molecular weight polyglycerol was used to synthesize novel stimuli-responsive hydrogels. Real-time monitoring of step-growth polymerization of glycerol was investigated using in-line and off-line Attenuated Total Reflectance/Fourier Transform infrared (ATR-FTIR) technique.

京都大学 / 新制・課程博士 / 博士(工学) / 甲第23226号 / 工博第4870号 / 新制||工||1760(附属図書館) / 京都大学大学院工学研究科高分子化学専攻 / (主査)教授 大内 誠, 教授 秋吉 一成, 教授 田中 一生 / 学位規則第4条第1項該当 / Doctor of Philosophy (Engineering) / Kyoto University / DGAM

京都大学 / 0048 / 新制・課程博士 / 博士(医科学) / 甲第21695号 / 医科博第99号 / 新制||医科||7(附属図書館) / 京都大学大学院医学研究科医科学専攻 / (主査)教授 生田 宏一, 教授 Shohab YOUSSEFIAN, 教授 齊藤 博英 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM