Thesis (M. App. Sc.)--University of Adelaide, Dept. of Mechanical Engineering, 1990. / Some mounted ill. Includes bibliographical references (leaves 48-50).

Thesis (M.Eng.Sc.) -- University of Adelaide, Dept. of Mechanical Engineering, 1983. / Typescript (photocopy).

The results of this work will be useful to those interested in predicting or improving the cutting accuracy of guided circular saws. In this thesis an experimentally verified numerical model of the guided fixed collar circular saw is presented. Features of the model include the ability to represent the blade and collar geometry, blade runout, rotational stresses, guide dynamics, guide pad shape, guide lubricant, and guide or blade misalignment. The blade is assumed to be governed by thin plate theory and the guide arms are modelled as a lumped parameter system. The lubricating fluid is modelled as a number of massless spring-dampers. Numerical solutions are given for the natural frequency response, the forced response due to static or harmonic lateral loading, and for the self-excited response caused by the interaction of the blade runout with the guides. The behaviour of the runout as a function of blade rotation speed and the conditions for which a resonant condition is produced in the guides are also determined. Experimental results obtained for the natural response, the deflection caused by a static load, the effect of speed on the blade runout, and the self-excited response correlate well with the numerical results. Numerical results are presented to show the effects of guide position, guide shape and the use of multiple guides on the natural and forced response. / Applied Science, Faculty of / Mechanical Engineering, Department of / Graduate

Made available in DSpace on 2014-06-11T19:31:00Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-08-19Bitstream added on 2014-06-13T20:21:35Z : No. of bitstreams: 1 vicente_ef_dr_araiq_parcial.pdf: 165362 bytes, checksum: 96b2cf2b3ae74363a4b5898b957a3b6a (MD5) Bitstreams deleted on 2015-06-03T11:42:42Z: vicente_ef_dr_araiq_parcial.pdf,. Added 1 bitstream(s) on 2015-06-03T11:44:08Z : No. of bitstreams: 1 000721605_20150819.pdf: 147542 bytes, checksum: f5cc64495e4f5a562a1e11d2ae711ce3 (MD5) Bitstreams deleted on 2015-08-20T11:58:40Z: 000721605_20150819.pdf,. Added 1 bitstream(s) on 2015-08-20T11:59:13Z : No. of bitstreams: 1 000721605.pdf: 4723399 bytes, checksum: 29bea79ab4fbb0c39fdce178c6b55872 (MD5) / A diidroorotato desidrogenase é uma enzima que apresenta um papel central na biossíntese de pirimidinas e catalisa a oxidação do diidroorotato a orotato. A enzima atua durante a via “de novo” de síntese de pirimidinas e está presente em quase todos os organismos vivos. A diidroorotato desidrogenase humana (HsDHODH) pode representar um importante alvo para o tratamento de doenças hiperproliferativas e inflamatórias, já que sua inibição bloqueia a síntese de ácidos nucléicos, impedindo a sua proliferação. Esta enzima tem uma estrutura monomérica e está associada com a membrana interna das mitocôndrias pela sua extensão N-terminal. Assim, entender em detalhes como esta enzima interage com a membrana poderia elucidar um alvo seletivo para drogas antiproliferativas, antiparasíticas e imunossupressivas. Esta região está também envolvida com a catálise central da enzima, sequestrando moléculas de ubiquinona presentes na membrana, fundamentais para as reações de oxirredução feitas pela enzima. Deste modo, para um melhor entendimento destes aspectos, neste trabalho foram sintetizados, por meio da Síntese de Peptídeos em Fase Sólida (SPFS) o peptídeo Ac-GDERFYAEHLMPTLQGLLDPESAHRLAVRFTSLG-NH2, que corresponde ao microdomínio existente na porção N-terminal da HsDHODH, entre os resíduos 33 e 66. Três análogos marcados com o aminoácido paramagnético TOAC nas posições 0, 12 ou 20, além de dois análogos duplamente marcados também foram obtidos. Ambos os peptídeos com dupla marcação possuem uma cisteína ligada ao spin MTSSL na posição 35 (C-terminal) se diferenciando pela posição do segundo marcador: um contendo outra cisteína ligada ao MTSSL na posição 12 e o segundo possuindo o TOAC na posição 0 (ou N-terminal). Estes peptídeos foram estudados por técnicas... / The dihydroorotate dehydrogenase is an enzyme that has a central role on the pyrimidine biosynthesis and catalyses the oxidation of dihydrorotate to orotate. The enzyme acts on de novo pyrimidines nucleotides pathway and it is present in almost all the live organisms. The human dihydroorotate dehydrogenase (HsDHODH) can represent an important target for the treatment of hiperproliferative and inflammatory diseases, since its inhibition blocks the nucleic acid synthesis, which restrains the cell proliferation. This enzyme has a monomeric structure and it is associated into the inner mitochondrial membrane by the N-terminal extension. Thus, understanding in details how this enzyme interacts with the membrane could help to elucidate a selective target for antiproliferative, antineoplasic and immunosuppressive drugs. This region is also involved with the central enzyme catalysis, harboring quinones molecules that are in the membranes, which is essential for the oxidation-reduction reactions made by the HsDHODH. In this way, for a better evaluation of these aspects, in this work we synthesized through the Solid Phase Peptide Synthesis (SPPS) the peptide Ac-GDERFYAEHLMPTLQGLLDPESAHRLAVRFTSLG-NH2, which corresponds to the HsDHODH N-terminal microdomain, between the residues 33 to 66. Three analogues labeled with the paramagnetic amino acid TOAC in the positions 0, 12 or 20 and two doubly labeled analogues were also synthesized. Both doubly labeled peptides contain a MTSSL-attached cysteine residue bounded to the position 35 (C-terminus), differing by the position of the second spin label: one possessing a cysteine with MTSSL at position 12 and the other contains TOAC at position 0 (N-terminus). These peptides were studied by spectroscopy techniques in order to obtain information about... (Complete abstract click electronic access below)

This thesis constructs special graphs with given circular chromatic numbers or circular flow numbers. Suppose $G=(V,E)$ is a graph and $rgeq 2$ is a real number. An $r$-coloring of a graph $G$ is a mapping $f:V ightarrow [0,r)$ such that for any adjacent vertices $x,y$ of $G$, $1leq |f(x)-f(y)|leq r-1$. The circular chromatic number $chi_c(G)$ is the least $r$ for which there exists an $r$-coloring of $G$. The circular chromatic number was introduced by Vince in 1988 in cite{vince}, where the parameter is called the {em star chromatic number} and denoted by $chi^*(G)$. Vince proved that for any rational number $k/dgeq 2$ there is a graph $G$ with $chi_c(G)=k/d$. In this thesis, we are interested in the existence of special graphs with given circular chromatic numbers. A graph $H$ is called a minor of a graph $G$ if $H$ can be obtained from $G$ by deleting some vertices and edges, and contracting some edges. A graph $G$ is called $H$-minor free if $H$ is not a minor of G. The well-known Hadwiger's conjecture asserts that for any positive integer $n$, any $K_n$-minor free graph $G$ is $(n-1)$-colorable. If this conjecture is true, then for any $K_n$-minor free graph $G$, we have $chi_c(G)leq n-1$. On the other hand, for any graph $G$ with at least one edge we have $chi_c(G)geq 2$. A natural question is this: Is it true that for any rational number $2leq rleq n-1$, there exist a $K_n$-minor free graph $G$ with $chi_c(G)=r$? For $n=4$, the answer is ``no". It was proved by Hell and Zhu in cite{hz98} that if $G$ is a $K_4$-minor free graph then either $chi_c(G)=3$ or $chi_c(G)leq 8/3$. So none of the rational numbers in the interval $(8/3,3)$ is the circular chromatic number of a $K_4$-minor free graph. For $ngeq 5$, Zhu cite{survey} proved that for any rational number $rin[2,n-2]$, there exists a $K_n$-minor free graph $G$ with $chi_c(G)=r$. The question whether there exists a $K_n$-minor free graph $G$ with $chi_c(G)=r$ for each rational number $rin(n-2,n-1)$ remained open. In this thesis, we answer this question in the affirmative. For each integer $ngeq 5$, for each rational number $rin[n-2,n-1]$, we construct a $K_n$-minor free graph $G$ with $chi_c(G)=r$. This implies that for each $ngeq 5$, for each rational number $rin[2,n-1]$, there exists a $K_n$-minor free graph $G$ with $chi_c(G)=r$. In case $n=5$, the $K_5$-minor free graphs constructed in this thesis are actually planar graphs. So our result implies that for each rational number $rin[2,4]$, there exists a planar graph $G$ with $chi_c(G)=r$. This result was first proved by Moser cite{moser} and Zhu cite{3-4}. To be precise, Moser cite{moser} proved that for each rational number $rin[2,3]$, there exist a planar graph $G$ with $chi_c(G)=r$, and Zhu cite{3-4} proved that for each rational number $rin[3,4]$, there exists a planar graph $G$ with $chi_c(G)=r$. Moser's and Zhu's proofs are quite complicated. Our construction is conceptually simpler. Moreover, for $ngeq 5$, $K_n$-minor free graphs, including the planar graphs are constructed with a unified method. For $K_4$-minor free graphs, although Hell and Zhu cite{hz98} proved that there is no $K_4$-minor free graph $G$ with $chi_c(G)in (8/3,3)$. The question whether there exists a $K_4$-minor free graph $G$ with $chi_c(G)=r$ for each rational number $rin[2,8/3]$ remained open. This thesis solves this problem: For each rational number $rin[2,8/3]$, we shall construct a $K_4$-minor free $G$ with $chi_c(G)=r$. This thesis also studies the relation between the circular chromatic number and the girth of $K_4$-minor free graphs. For each integer $n$, the supremum of the circular chromatic number of $K_4$-minor free graphs of odd girth (the length of shortest odd cycle) at least $n$ is determined. It is also proved that the same bound is sharp for $K_4$-minor free graphs of girth $n$. By a classical result of ErdH{o}s, for any positive integers $l$ and $n$, there exists a graph $G$ of girth at least $l$ and of chromatic number $n$. Using probabilistic method, Zhu cite{unique} proved that for each integer $l$ and each rational number $rgeq 2$, there is a graph $G$ of girth at least $l$ such that $chi_c(G)=r$. Construction of such graphs for $rgeq 3$ was given by Nev{s}etv{r}il and Zhu cite{nz}. The question of how to construct large girth graph $G$ with $chi_c(G)=r$ for given $rin(2,3)$ remained open. In this thesis, we present a unified method that constructs, for any $rgeq 2$, a graph $G$ of girth at least $l$ with circular chromatic number $chi_c(G) =r$. Graphs $G$ with $chi_c(G)=chi(G)$ have been studied extensively in the literature. Many families of graphs $G$ are known to satisfy $chi_c(G)=chi(G)$. However it remained as an open question as how to construct arbitrarily large $chi$-critical graphs $G$ of bounded maximum degree with $chi_c(G)=chi(G)$. This thesis presents a construction of such graphs. The circular flow number $Phi_c(G)$ is the dual concept of $chi_c(G)$. Let $G$ be a graph. Replace each edge $e=xy$ by a pair of opposite arcs $a=overrightarrow{xy}$ and $a^{-1}=overrightarrow{yx}$. We obtain a symmetric directed graph. Denote by $A(G)$ the set of all arcs of $G$. A chain is a mapping $f:A(G) ightarrow I!!R$ such that for each arc $a$, $f(a^{-1})=-f(a)$. A flow is a chain such that for each subset $X$ of $V(G)$, $sum_{ain[X,ar{X}]}f(a)=0$, where $[X,ar{X}]$ is the set of all arcs from $X$ to $V-X$. An $r$-flow is a flow such that for any arc $ain A(G)$ , $1leq |f(a)| leq r-1$. The circular flow number of $G$ is $Phi_c(G)=mbox{ inf}{r: G mbox{ admits a } rmbox{-flow}}$. It was conjectured by Tutte that every graph $G$ has $Phi_c(G)leq 5$. By taking the geometrical dual of planar graphs, Moser's and Zhu's results concerning circular chromatic numbers of planar graphs imply that for each rational number $rin[2,4]$, there is a graph $G$ with $Phi_c(G)=r$. The question remained open whether for each $rin(4,5)$, there exists a graph $G$ with $Phi_c(G)=r$. In this thesis, for each rational number $rin [4,5]$, we construct a graph $G$ with $Phi_c(G)=r$.

Made available in DSpace on 2014-06-11T19:27:21Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-02-13Bitstream added on 2014-06-13T20:56:17Z : No. of bitstreams: 1 goncalesgarcia_l_me_sjrp.pdf: 353281 bytes, checksum: 6786716aa7970dd6c603e25cad2d5709 (MD5) / As atividades realizadas compreenderam a produção de cDNA utilizando primer anti-senso e RNA purificado, amplificação do gene codificador da proteína 25K do Cole latent virus (CoLV25K), purificação do fragmento amplificado, ligação em vetor de multiplicação pGEM-T, transformação em células competentes de Escherichia coli linhagem TOP 10, purificação do vetor, digestão enzimática com as enzimas Bam HI e Hind III, subclonagem no vetor de expressão pET28a, transformação em células competentes de E. coli linhagem BL21-RIL, sequenciamento do gene no vetor de expressão e expressão da proteína a 37 o C . Quando expressa a 37 ºC, a proteína, de 25 kDa, foi encontrada em sua totalidade nos corpos de inclusão. Dessa forma, a proteína foi purificada sob condição desnaturante (utilizando 8 M de uréia) e submetida à diálise para seu reenovelamento. Após o reenovelamento, a proteína foi concentrada para aproximadamente 3 mg/mL e foram realizadas medidas de dicroísmo circular, para verificar o seu conteúdo de estrutura secundária, e o espalhamento dinâmico de luz, para estimar a distribuição de tamanho das populações de partículas que estão presentes na solução. Os dados da deconvolução do experimento de dicroísmo circular indicam um percentual de 40-46% α-hélice, 12-14% folha-β, 15-22% voltas e 24-28% de outras estruturas, indicando que a proteína está estruturada; e os dados do espalhamento dinâmico de luz mostraram que a proteína encontra-se estável, monodispersa, mas apresenta um complexo de partículas que deve ser removido para que fique nas condições ideais de cristalização. Foram utilizadas também outras técnicas para tentar alcançar a solubilidade da proteína expressa: abaixando a temperatura... / The experiments performed were, the production of cDNA using anti-sense primers and purified RNA, amplification of the gene encoding the 25K protein of Cole latent virus (CoLV25K), purification of the amplified fragment, cloning in multiplication vector pGEM-T for cell transformation into competent Escherichia coli strain TOP 10, vector purification, enzymatic digestion using the enzymes Bam HI and Hind III, subcloning in expression vector pET28a, transformation into competent cells of E. coli strain BL21-RIL, sequencing of the gene cloned into the expression vector and protein expression at 37 o C. When expressed at 37 °C, the protein with a molecular mass of 25 kDa was detected in inclusion bodies. Thus, the protein was purified under denaturing conditions (using 8 M urea) and subjected to dialysis to stimulate refolding. After refolding, the protein was concentrated to approximately 3 mg / mL and the circular dichroism assay was performed to verify the content of secondary structure, and dynamic light scattering, to estimate the size distribution of particle populations which are present in solution. The data from the deconvolution of circular dichroism experiments indicate a percentage of 40-46% α-helix, 12-14% β-sheet, 15-22% turns and 24-28% of other structures, indicating a structured protein; and the data of the dynamic light scattering showed that the protein is stable, monodispersive, but forms a large complex of particles which must be separated to before crystallization experiments. Other techniques were used to solubilize the expressed protein: lowering the temperature of expression to 18 °C, using the method... (Complete abstract click electronic access below)

Orientador: Eduardo Maffud Cilli / Banca: José Roberto Ernandes / Banca: Patricia Targon Campana / Banca: Antonio José da Costa Filho / Banca: Vani Xavier de Oliveira Junior / Resumo: A diidroorotato desidrogenase é uma enzima que apresenta um papel central na biossíntese de pirimidinas e catalisa a oxidação do diidroorotato a orotato. A enzima atua durante a via "de novo" de síntese de pirimidinas e está presente em quase todos os organismos vivos. A diidroorotato desidrogenase humana (HsDHODH) pode representar um importante alvo para o tratamento de doenças hiperproliferativas e inflamatórias, já que sua inibição bloqueia a síntese de ácidos nucléicos, impedindo a sua proliferação. Esta enzima tem uma estrutura monomérica e está associada com a membrana interna das mitocôndrias pela sua extensão N-terminal. Assim, entender em detalhes como esta enzima interage com a membrana poderia elucidar um alvo seletivo para drogas antiproliferativas, antiparasíticas e imunossupressivas. Esta região está também envolvida com a catálise central da enzima, sequestrando moléculas de ubiquinona presentes na membrana, fundamentais para as reações de oxirredução feitas pela enzima. Deste modo, para um melhor entendimento destes aspectos, neste trabalho foram sintetizados, por meio da Síntese de Peptídeos em Fase Sólida (SPFS) o peptídeo Ac-GDERFYAEHLMPTLQGLLDPESAHRLAVRFTSLG-NH2, que corresponde ao microdomínio existente na porção N-terminal da HsDHODH, entre os resíduos 33 e 66. Três análogos marcados com o aminoácido paramagnético TOAC nas posições 0, 12 ou 20, além de dois análogos duplamente marcados também foram obtidos. Ambos os peptídeos com dupla marcação possuem uma cisteína ligada ao spin MTSSL na posição 35 (C-terminal) se diferenciando pela posição do segundo marcador: um contendo outra cisteína ligada ao MTSSL na posição 12 e o segundo possuindo o TOAC na posição 0 (ou N-terminal). Estes peptídeos foram estudados por técnicas... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The dihydroorotate dehydrogenase is an enzyme that has a central role on the pyrimidine biosynthesis and catalyses the oxidation of dihydrorotate to orotate. The enzyme acts on "de novo" pyrimidines nucleotides pathway and it is present in almost all the live organisms. The human dihydroorotate dehydrogenase (HsDHODH) can represent an important target for the treatment of hiperproliferative and inflammatory diseases, since its inhibition blocks the nucleic acid synthesis, which restrains the cell proliferation. This enzyme has a monomeric structure and it is associated into the inner mitochondrial membrane by the N-terminal extension. Thus, understanding in details how this enzyme interacts with the membrane could help to elucidate a selective target for antiproliferative, antineoplasic and immunosuppressive drugs. This region is also involved with the central enzyme catalysis, harboring quinones molecules that are in the membranes, which is essential for the oxidation-reduction reactions made by the HsDHODH. In this way, for a better evaluation of these aspects, in this work we synthesized through the Solid Phase Peptide Synthesis (SPPS) the peptide Ac-GDERFYAEHLMPTLQGLLDPESAHRLAVRFTSLG-NH2, which corresponds to the HsDHODH N-terminal microdomain, between the residues 33 to 66. Three analogues labeled with the paramagnetic amino acid TOAC in the positions 0, 12 or 20 and two doubly labeled analogues were also synthesized. Both doubly labeled peptides contain a MTSSL-attached cysteine residue bounded to the position 35 (C-terminus), differing by the position of the second spin label: one possessing a cysteine with MTSSL at position 12 and the other contains TOAC at position 0 (N-terminus). These peptides were studied by spectroscopy techniques in order to obtain information about... (Complete abstract click electronic access below) / Doutor

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