Quaternary centres bearing a nitrogen atom are found in natural products and therapeutic agents but they represent a remarkably challenging synthetic motif to access when stereochemical control is required. This thesis details investigations into the development of an innovative approach that - by combining biocatalysis with organolithium chemistry - allows the synthesis of enantioenriched α-tertiary amines. The strategy relies on the initial enzymatic asymmetric synthesis of amines. Two complementary pathways were identified: deracemisation with amine oxidases or enantioselective reduction with imine reductases. The enantioenriched amines were converted to the corresponding N-benzyl-N'-aryl ureas and subsequent organolithium-mediated stereospecific aryl migration developed in the Clayden group were carried out to obtain α-tertiary amines. Various scaffold were investigated: 1,1-disubstituted 1,2,3,4-tetrahydroisoquinoline, 2,2-disubstituted azepane and 1,1-disubstituted 2,3,4,5-tetrahydro-1H-benzo[c]azepine derivatives were successfully synthesised. The methodology was extended to acyclic systems, giving 3-pyridyl-derived α-tertiary amines.

The non-increasing rearrangement of a measurable real function defined on an appropriate measure space is of the enormous significance in disciplines such as theory of function spaces or interpolation theory and their applications in PDEs. Unfortunately, while it has good and widely applicable mapping properties, it is virtually impossible to calculate the non-increasing rearrangement of a concrete given function precisely. Numerical algorithms for approximation are desirable for this reason. Such method of approximation, based on interpolation by a linear spline, is presented in this thesis. In the first half of this thesis, the developed method is described, while the error estimates of the method are subject to the second part.

Nesta dissertação, investigarei a configuração do quadro partidário brasileiro pós-revolução de 30 e busco levantar explicações sobre o porquê deste rearranjo partidário. Estas explicações foram levantadas com base em dois elementos explicativos encontrados na literatura, que corresponde: a explicação institucionalista e a contingencial e em cima destas será elaborada a resposta sobre o problema em tela. Este trabalho ao dar atenção a este tema traz luz a um problema ainda não investigado pela ciência política brasileira e abre um leque para a emergência de uma nova agenda de pesquisa na Ciência Política brasileira. / In this dissertation I will investigate the Brazilian party framework configuration in the period after the revolution of 1930 and I aim to seek for explanations about this party rearrangement. These explanations were raised in connection with two explanatory elements found in the literature: the institutionalism and contingency explanations, over these two theories the answer to the research problem will be given. This work pretends to give attention to a very important problem that wasnt investigated by Brazilian political science and consequently this will open a new research agenda inside Brazilian political science.

Alcaloides indolizidínicos, quinolizidínicos e piperidínicos poliidroxilados representam classes de compostos amplamente investigados atualmente. Este fato se deve às pronunciadas atividades biológicas como inibidores de glicosidases expressadas por estes heterociclos, o que significa um grande atrativo para que muitos grupos de pesquisas desenvolvam metodologias sintéticas para a sua obtenção de forma efetiva e em poucas etapas. Neste trabalho de dissertação é apresentada uma rota sintética para a preparação de alcaloides indolizidínicos e quinolizidínicos diidroxilados a partir de diazocetonas α,β-insaturadas. A estratégia para a síntese destes alcaloides baseia-se na mesma metodologia, tendo como etapas chaves: uma reação de olefinação de Horner-Wadsworth-Emmons (HWE) a partir de aminoaldeídos, seguida de um Rearranjo de Wolff. Como material de partida para o esqueleto indolizidínico foi empregado o Cbz-S-prolinal. Este aminoaldeído foi empregado como fonte do centro estereogênico e de um dos anéis presentes na estrutura final. O acoplamento (reação de HWE) entre o Cbz-S-prolinal e um diazofosfonato recentemente descrito por nosso grupo de pesquisa forneceu um composto diazocarbonílico α,β-insaturado (67%), que em seguida, foi submetido a um rearranjo de Wolff fornecendo um éster β,γ-insaturado (96%). Este intermediário avançado foi funcionalizado através de uma reação de diidroxilação, a qual forneceu uma lactona (66%). A síntese foi completada através de uma reação de ciclização intramolecular (94%) seguida de uma reação de redução para fornecer a indolizidina diidroxilada em 71% de rendimento. Para o esqueleto quinolizidínico, foi empregado o aminoaldeído racêmico Cbz-(±)-pipecolinal como material de partida. A partir da reação de olefinação, foi obtida uma diazocetona α,β-insaturada (91%), que após Rearranjo de Wolff (95%), diidroxilação (75%), ciclização (54-74%) e reação de redução (87-90%), forneceu duas novas quinolizidinas diidroxiladas. Estes alcaloides indolizidínicos e quinolizidínicos poderão ser avaliados como inibidores de glicosidases. / Polyhydroxylated Indolizidine, quinolizidine and piperidine alkaloids represent classes of compounds widely investigated in the chemical community. This fact is due to their pronounced biological activities as glycosidase inhibitors. Considering that, many research groups have been developing new synthetic methodologies to obtain these alkaloids and analogs effectively and in few steps. This work presents a synthetic route for the preparation of dihydroxylated indolizidine and quinolizidine alkaloids from α,β-unsaturated diazoketones. The strategy for the synthesis of these compounds is based on the same methodology to construct the indolizidine and quinolizidine skeleton. The key steps involve a Horner-Wadsworth-Emmons (HWE) olefination reaction from aminoaldehydes, followed by a Wolff Rearrangement. As the starting material to construct the indolizidine skeleton, Cbz-S-prolinal was employed. This aminoaldehyde is also the source of the stereogenic center and one of the rings present in the final structure. The coupling reaction (HWE reaction) between Cbz-S-prolinal and a diazophosphonate (methodology recently described by our research group) has provided an α,β-unsaturated diazoketone in 67% yield. This compound was then subjected to a Wolff Rearrangement, providing a β,γ-unsaturated ester in 96% yield. This advanced intermediate was functionalized through a high selective dihydroxylation reaction, furnishing a hydroxylated lactone in 66% yield. The synthesis was then completed employing an intramolecular cyclization reaction (94% yield), followed by lactam reduction to provide the dihydroxylated indolizidine (1,6-dideoxyepicastanospermine) in 71% yield. For the construction of the quinolizidine skeleton, was employed racemic Cbz-pipecolinal as the starting material. From the olefination reaction, the corresponding α,β-unsaturated diazoketone was obtained in 91% yield. After a Wolff Rearrangement (95%), dihydroxylation reaction (75%), cyclization (54-74%) and lactam reduction (87-90%), two novel dihydroxylated quinolizidines could be synthesized. These indolizidine and quinolizidine alkaloids may be evaluated as new inhibitors of glycosidases.

<p>Bacteria of the genus <i>Bartonella</i> inhabit the red blood cells of many mammals, including humans, and are transmitted by blood-sucking arthropod vectors. Different species of <i>Bartonella</i> are associated with different mammalian host species, to which they have adapted and normally do not cause any symptoms. Incidental infection of other hosts is however often followed by various disease symptoms, and several <i>Bartonella</i> species are considered as emerging human pathogens.</p><p>In this work, I have studied the genomic diversity within and between different <i>Bartonella</i> species, with focus on the feline-associated human pathogen <i>B. henselae</i> and its close relatives, the similarly feline-associated <i>B. koehlerae</i> and the trench-fever agent <i>B. quintana</i> which is restricted to humans.</p><p>In <i>B. henselae</i>, the overall variability in sequence and genome content was modest and well correlated, suggesting low levels of intra-species recombination in the core genome. The variably present genes were located in the prophage and the genomic islands, which are also absent from <i>B. quintana</i> and <i>B. koehlerae</i>, indicating multiple independent excision events. In contrast, diversity of genome structures was immense and probably associated with rearrangements between the repeated genomic islands located around the terminus of replication, possibly to avoid the host’s immune system. In both <i>B. henselae</i> and the mouse-associated species <i>B. grahamii</i> a large portion of the chromosome was manifold amplified in long-time cultures and packaged into phage particles, allowing for different recombination rates for different chromosomal regions.</p><p>In B<i>. quintana</i>, diversity was studied by sequencing non-coding spacers. The low variability might be due to the recent emergence of this species. Surprisingly, also this species displayed high variability in genome structures, despite its lack of repeated sequences.</p><p>The results indicate that genome rearrangements and gain or loss of mobile elements are major mechanisms of evolution in <i>Bartonella</i>.</p>

Bacteria of the genus Bartonella inhabit the red blood cells of many mammals, including humans, and are transmitted by blood-sucking arthropod vectors. Different species of Bartonella are associated with different mammalian host species, to which they have adapted and normally do not cause any symptoms. Incidental infection of other hosts is however often followed by various disease symptoms, and several Bartonella species are considered as emerging human pathogens. In this work, I have studied the genomic diversity within and between different Bartonella species, with focus on the feline-associated human pathogen B. henselae and its close relatives, the similarly feline-associated B. koehlerae and the trench-fever agent B. quintana which is restricted to humans. In B. henselae, the overall variability in sequence and genome content was modest and well correlated, suggesting low levels of intra-species recombination in the core genome. The variably present genes were located in the prophage and the genomic islands, which are also absent from B. quintana and B. koehlerae, indicating multiple independent excision events. In contrast, diversity of genome structures was immense and probably associated with rearrangements between the repeated genomic islands located around the terminus of replication, possibly to avoid the host’s immune system. In both B. henselae and the mouse-associated species B. grahamii a large portion of the chromosome was manifold amplified in long-time cultures and packaged into phage particles, allowing for different recombination rates for different chromosomal regions. In B. quintana, diversity was studied by sequencing non-coding spacers. The low variability might be due to the recent emergence of this species. Surprisingly, also this species displayed high variability in genome structures, despite its lack of repeated sequences. The results indicate that genome rearrangements and gain or loss of mobile elements are major mechanisms of evolution in Bartonella.

Among Nature’s creation, terpenoids are more versatile and exciting natural products. In a remarkable display of synthetic ingenuity and creativity, nature has endowed terpenes with a bewildering array of carbocyclic frameworks with unusual assemblage of rings and functionalities. This phenomenal structural diversity of terpenes make them ideal targets for developing and testing new synthetic strategies for efficient articulation of carbocyclic frameworks. The thesis entitled “Total synthesis of sesquiterpenes acorenols, chamigrenes, and laurokamurene B; and Enantiospecific synthesis of ABC-ring system of A-nor and abeo pentacyclic triterpenes” describes the studies directed towards the total synthesis of the sesquiterpenes mentioned in the title and exploratory studies towards triterpenoids. In each chapter of the thesis, the compounds are sequentially numbered (bold) and references are marked sequentially as superscripts and listed at the end of the chapter. All the spectra included in the thesis were obtained by xeroxing the original NMR spectra. The sesquiterpenes acorenols, containing an interesting spiro[4.5]decane carbon framework, was first isolated in 1970 by the research group of Tomita from the wood of Juniperus rigida. Recently, in 2003, Braun and coworkers reported the isolation of epi α- and epi β-acorenols along with α- and β-acorenols from the sandal wood oil Santalum spicatum. Total synthesis of all the four acorenols has been described in the first part of the first chapter of the thesis. Initially, a model study has been carried out for the spirocyclopentannulation of cyclohexanone employing a combination of Ireland ester Claisen rearrangement and ring closing metathesis reaction to furnish methyl 4-methylspiro[4.5]dec-3-en-1-carboxylate. The same methodology has been extended for the total synthesis of all the four acorenols starting from cyclohexane-1,4-dione via cis and trans isomers of methyl 4-methyl-8-methylene-spiro[4.5]dec-3-ene-1-carboxylate. Total synthesis of β-chamigrene, γ-chamigrene and laurencenone C, containing spiro[5.5]undecane carbon framework, has been described in the second part of the first chapter. As a model study, cyclohexanone has been transformed into 1,5,5-trimethylspiro-[5.5]undec-4-en-3-one employing a combination of Ireland ester Claisen rearrangement and intramolecular type-II carbonyl ene reactions. The methodology has been extended to chamigrenes starting from cyclohexane-1,4-dione via methyl 2-(1-isopropenyl-4-oxocyclo-hexyl)-2-methylpropanoate and 5,5-dimethyl-1,9-ismethylenespiro[5.5]undecan-3-ol. The marine sesquiterpenes laurokamurenes were first isolated in 2006 by Mao and Guo from Laurencia okamurai Yamada. First total synthesis of (±)-laurokamurene B has been described in the first part of the second chapter. To begin with Ireland ester Claisen rearrangement of but-2-enyl 2-methylpropionate furnished methyl 2,2,3-trimethylpent-4-enoate, which was then transformed into 4,5,5-trimethyl-3-(4-methylphenyl)hepta-1,6-dien-3-ol. RCM reaction followed by reductive deoxygeneation transformed 4,5,5-trimethyl-3-(4-methylphenyl)hepta-1,6-dien-3-ol into (±)-laurokamurene B. Subsequently, an enantioselective total synthesis of (+)-laurokamurene B has been accomplished. Stereoselective hydrogenation of methyl campholenoate furnished methyl 2-[(1S,3S)-2,2,3-trimethyl-cyclopent-1-yl]acetate, which was then transformed into (+)-laurokamurene B via degradation of the two carbon side chain and introduction of the aryl moiety, which established the absolute configuration of laurokamurenes. The third chapter deals with the enantiospecific generation of ABC-ring system of A-nor and abeo 4(3 → 2) tetra and pentacyclic triterpenes. To begin with (R)-carvone was identified as B-ring of ABC-ring system of A-nor and abeo tetra and pentacyclic triterpenes, as the absolute configuration at the C-5 position of the targets correlate to the stereo centre of carvone, and isopropenyl group can serve as the C-4 carbon of the targets along with the two gem-dimethyl groups. A lithium liquid ammonia mediated cyclisation of δ,ε-unsaturated esters was employed for the construction of the A ring and an RCM reaction was opted for the construction of the C ring. (R)-Carvone has been converted into 2-(1-ethoxyethoxy)-1,3,7,7-tetramethylbicyclo[4.3.0]non-3-en-8-ol via lithium liquid ammonia mediated cyclisation of methyl 2-(1-ethoxyethoxy)-6-isopropenyl-1,3-dimethylcyclohex-3-enyl]acetate, which was then transformed into 4-methoxymethoxy-2,5,5,9-tetramethyltricyclo[7.4.0.02,6]tridec-11-en-8-one via the RCM reaction of 3,4-bisallyl-8-methoxymethoxy-4,6,9,9-tetramethylbicyclo-[4.3.0]nonan-3-one. The strategy has been further extended to the synthesis of 4-methylene-2,5,5,9-tetramethyltricyclo[7.4.0.02,6]tridec-11-en-8-one, which contains the ABC ring system of abeo 4(3→2) tetra and pentacyclic triterpenes.

In Sweden, close to 10 000 men are annually diagnosed with prostate cancer (PCa) and approximately 2400 men die of their disease each year. Today there is no reliable marker that can separate patients who will have an aggressive type of disease that requires treatment, from patients who will have a more indolent clinical course and can be left untreated. This further leads to the current problem of over treatment of men with PCa. Hence, there is an urgent need for reliable prognostic markers that can be used at time of diagnosis. With the discovery of recurrent gene rearrangements in PCa, most commonly ERG rearrangements, hope came that this aberration could play a role in diagnosis and/or prognosis of the disease. The aim of this thesis was to investigate the clinical implication of ERG rearrangements in the management of PCa. The work in this thesis supports the findings from previous studies, suggesting that the ERG rearrangement is a sign of a more aggressive type of cancer. The major findings are that in multifocal PCa, the ERG rearranged cancer foci are more prone to metastatic dissemination compared to foci without the ERG rearrangement and that patients harboring the ERG rearrangement have a faster disease progression leading up to earlier start of hormonal treatment. Furthermore, the results add an additional level of complexity in a subset of PCa tumors that harbor multiple gene rearrangements on the cellular level. The result also show that the newly available ERG antibody is highly predictive of ERG rearrangement and is appropriate to use when faced with limitations in tissue amounts. The findings in this thesis indicate that the ERG rearrangement has a potential role in the clinical management of PCa but further studies arerequired.

Genomic rearrangements, particularly deletions and duplications, are known to cause many genetic disorders. The chromosome 7q11.23 region in humans is prone to recurrent chromosomal rearrangement, due to the presence of low copy repeats that promote non-allelic homologous recombination. The most well characterized rearrangement of 7q11.23 is a hemizygous 1.5 million base pair (Mb) deletion spanning more than 25 genes. This deletion causes Williams-Beuren Syndrome (WBS; OMIM 194050), a multisystem developmental disorder with distinctive physical and behavioural features. Other rearrangements of the region lead to phenotypes distinct from that of WBS. Here we describe the first individual identified with duplication of the same 1.5 Mb region, resulting in severe impairment of expressive language, in striking contrast to people with WBS who have relatively well preserved language skills. We also describe the identification of a new gene for a severe form of childhood epilepsy through the analysis of individuals with deletions on chromosome 7 that extend beyond the boundaries typical for WBS. This gene, MAGI2, is part of the large protein scaffold at the post-synaptic membrane and provides a new avenue of research into both the molecular basis of infantile spasms and the development of effective therapies. Individuals with smaller than typical deletions of 7q11.23 have delineated a minimal critical region for WBS and have implicated two members of the TFII-I transcription factor family. To better understand the contribution of these genes to WBS, I have generated animal models with these genes deleted singly and in combination. Disruption of the first gene, Gtf2ird1, resulted in phenotypes reminiscent of WBS including alterations in social behaviour, natural fear response and anxiety. An alteration in serotonin function was identified in the frontal cortex and may be linked to these behavioural phenotypes. Together with a model for the second gene, Gtf2i, and the double deletion model that was generated using Cre-loxP technology, these resources will permit the study of the individual and additive effects of hemizygosity for Gtf2i and Gtf2ird1 and will greatly expand our understanding of the role the TFII-I gene family in WBS.

Genomic rearrangements, particularly deletions and duplications, are known to cause many genetic disorders. The chromosome 7q11.23 region in humans is prone to recurrent chromosomal rearrangement, due to the presence of low copy repeats that promote non-allelic homologous recombination. The most well characterized rearrangement of 7q11.23 is a hemizygous 1.5 million base pair (Mb) deletion spanning more than 25 genes. This deletion causes Williams-Beuren Syndrome (WBS; OMIM 194050), a multisystem developmental disorder with distinctive physical and behavioural features. Other rearrangements of the region lead to phenotypes distinct from that of WBS. Here we describe the first individual identified with duplication of the same 1.5 Mb region, resulting in severe impairment of expressive language, in striking contrast to people with WBS who have relatively well preserved language skills. We also describe the identification of a new gene for a severe form of childhood epilepsy through the analysis of individuals with deletions on chromosome 7 that extend beyond the boundaries typical for WBS. This gene, MAGI2, is part of the large protein scaffold at the post-synaptic membrane and provides a new avenue of research into both the molecular basis of infantile spasms and the development of effective therapies. Individuals with smaller than typical deletions of 7q11.23 have delineated a minimal critical region for WBS and have implicated two members of the TFII-I transcription factor family. To better understand the contribution of these genes to WBS, I have generated animal models with these genes deleted singly and in combination. Disruption of the first gene, Gtf2ird1, resulted in phenotypes reminiscent of WBS including alterations in social behaviour, natural fear response and anxiety. An alteration in serotonin function was identified in the frontal cortex and may be linked to these behavioural phenotypes. Together with a model for the second gene, Gtf2i, and the double deletion model that was generated using Cre-loxP technology, these resources will permit the study of the individual and additive effects of hemizygosity for Gtf2i and Gtf2ird1 and will greatly expand our understanding of the role the TFII-I gene family in WBS.