Block co-polymerization by transformation reactions

The aim of this study was to use the transformation of anionic to metathesis polymerization to produce block co-polymers of styrene-b-pentenylene using WC16 /PStLi and WC16/PStLi/ AlEtC12 catalyst systems. Analysis of the products using SEC and 1H and 13C NMR spectroscopy enabled mechanisms for metathesis initiation reactions to be proposed. The initial work involved preparation of the constituent homo-polymers. Solutions of polystyryllithium in cyclohexane were prepared and diluted so that the [PStLi]o<2x10-3M. The dilution produced initial rapid decay of the active species, followed by slower spontaneous decay within a period of days. This was investigated using UV / visible spectrophotometry and the wavelength of maximum absorbance of the PStLi was found to change with the decay from an initial value of 328mn. to λmax of approximately 340nm. after 4-7 days. SEC analysis of solutions of polystyrene, using RI and UV / visible (set at 254nm.) detectors; showed the UV:RI peak area was constant for a range of polystyrene samples of different moleculor weight. Samples of polypentenylene were prepared and analysed using SEC. Unexpectedly the solutions showed an absorbance at 254nm. which had to be considered when this technique was used subsequently to analyse polymer samples to determine their styrene/ pentenylene co-polymer composition. Cyclohexane was found to be a poor solvent for these ring-opening metathesis polymerizations of cyclopentene. Attempts to produce styrene-b-pentenylene block co-polymers, using a range of co-catalyst systems, were generally unsuccessful as the products were shown to be mainly homopolymers. The character of the polymers did suggest that several catalytic species are present in these systems and mechanisms have been suggested for the formation of initiating carbenes. Evidence of some low molecular weight product with co-polymer character has been obtained. Further investigation indicated that this is most likely to be ABA block copolymer, which led to a mechanism being proposed for the termination of the polymerization.

Functional characterization of two human receptor activity-modifying protein 3 variants

Adrenomedullin (AM) and amylin are involved in angiogenesis/lymphangiogenesis and glucose homeostasis/food intake, respectively. They activate receptor activity-modifying protein (RAMP)/G protein-coupled receptor (GPCR) complexes. RAMP3 with the calcitonin receptor-like receptor (CLR) forms the AM(2) receptor, whereas when paired with the calcitonin receptor AMY(3) receptors are formed. RAMP3 interacts with other GPCRs although the consequences of these interactions are poorly understood. Therefore, variations in the RAMP3 sequence, such as single nucleotide polymorphisms or mutations could be relevant to human health. Variants of RAMP3 have been identified. In particular, analysis of AK222469 (Homo sapiens mRNA for receptor (calcitonin) activity-modifying protein 3 precursor variant) revealed several nucleotide differences, three of which encoded amino acid changes (Cys40Trp, Phe100Ser, Leu147Pro). Trp56Arg RAMP3 is a polymorphic variant of human RAMP3 at a conserved amino acid position. To determine their function we used wild-type (WT) human RAMP3 as a template for introducing amino acid mutations. Mutant or WT RAMP3 function was determined in Cos-7 cells with CLR or the calcitonin receptor (CT((a))). Cys40Trp/Phe100Ser/Leu147Pro RAMP3 was functionally compromised, with reduced AM and amylin potency at the respective AM(2) and AMY(3(a)) receptor complexes. Cys40Trp and Phe100Ser mutations contributed to this phenotype, unlike Leu147Pro. Reduced cell-surface expression of mutant receptor complexes probably explains the functional data. In contrast, Trp56Arg RAMP3 was WT in phenotype. This study provides insight into the role of these residues in RAMP3. The existence of AK222469 in the human population has implications for the function of RAMP3/GPCR complexes, particularly AM and amylin receptors.

Delivery and stability of antisense oligonucleotide conjugates in vitro

The efficacy of antisense oligonucleotide (ODN) therapy is dependent on four major parameters: delivery to cells, intracellular stability and localisation and efficient action at the target site.The aim of this project was to study the delivery of ODNs to macrophages and to assess the stability of two ODN conjugates, in vitro. The first conjugate aimed to improve uptake of ODNs via mannose receptor mediated delivery, the second investigated the improved delivery of ODN conjugates via non-specific lipophilic interaction with the cell membrane. A mono-mannose phosphoramidite derivative was designed and synthesised and a mono-mannose ODN conjugate synthesised by standard phosphoramidite chemistry. Delivery of this conjugate was enhanced to RAW264.7 and J774 macrophage cell lines via a mechanism of receptor mediated endocytosis. The delivery of three lipophilic ODN conjugates, cholesterol (cholhex), 16-carbon alkyl chain (C16) and hexa-ethylene glycol (HEG) moieties and an unconjugated ODN were assessed in RAW264.7 macrophages. All three conjugates increased the lipophilicity of the ODN as assessed from partition coefficient data. Both the cholhex and unconjugated ODNs were found to have higher degrees of cellular association than the C16 and HEG conjugates. Cellular uptake studies implicated internalisation of these ODNs by an adsorptive endocytosis mechanism. Following endocytosis, ODNs must remain stable during their residence in endosomal/lysosomal compartments prior to exiting and exerting their biological action in either the cytosol or nucleus. Assessment of in vitro stability in a lysosomal extract revealed the cholhex conjugate and unconjugated ODNs to have a longer half-life than the C16 and HEG conjugated ODNs, highlighting the influence of conjugate moieties on lysosomal stability. The effects of base composition and length on stability in a lysosomal extract revealed the longest half-life for homo-cytidine ODNs and ODNs over 20 nucleotides in length. These studies suggest that the above conjugates can enhance cellular association and delivery of antisense ODNs to cultured macrophages. This may lead to their use in treating disorders such as HIV infection, which affects this cell type.

Intolerance of ambiguity, gender stereotypes, and attitudes to sexuality

The premise of this thesis is that Western thought is characterised by the need to enforce binary classification in order to structure the world. Classifications of sexuality and gender both embody this tendency, which has been largely influenced by Judeo-Christian tradition. Thus, it is argued that attitudes to sexuality, particularly homosexuality are, in part, a function of the way in which we seek to impose structure on the world. From this view, it is (partly) the ambiguity, inherent in gender and sexual variation, which evokes negative responses. The thesis presents a series of inter-linked studies examining attitudes to various aspects of human sexuality, including the human body, non-procreative sex acts (anal an oral sex) and patterns of sexuality that depart from the hetero-homo dichotomy. The findings support the view that attitudes to sexuality are significantly informed by gender-role stereotypes, with negative attitudes linked to intolerance of ambiguity. Male participants show large differences in their evaluations of male and female bodies, and of male and female sexual actors, than do female participants. Male participants also show a greater negativity to gay male sexual activity than do female participants, but males perceive lesbian sexuality similarly to heterosexuality. Male bodies are rated as being less 'permeable' than female bodies and male actors are more frequently identified as being the instigators of sexual acts. Crucial to the concept of heterosexism is the assumption that 'femininity' is considered inherently inferior to 'masculinity'. Hence, the findings provide an empirical basis for making connections between heterosexism and sexism, and therefore between the psychology of women, and gay and lesbian psychology.

Common variants in left/right asymmetry genes and pathways are associated with relative hand skill

Humans display structural and functional asymmetries in brain organization, strikingly with respect to language and handedness. The molecular basis of these asymmetries is unknown. We report a genome-wide association study meta-analysis for a quantitative measure of relative hand skill in individuals with dyslexia [reading disability (RD)] (n = 728). The most strongly associated variant, rs7182874 (P = 8.68×10-9), is located in PCSK6, further supporting an association we previously reported. We also confirmed the specificity of this association in individuals with RD; the same locus was not associated with relative hand skill in a general population cohort (n = 2,666). As PCSK6 is known to regulate NODAL in the development of left/right (LR) asymmetry in mice, we developed a novel approach to GWAS pathway analysis, using gene-set enrichment to test for an over-representation of highly associated variants within the orthologs of genes whose disruption in mice yields LR asymmetry phenotypes. Four out of 15 LR asymmetry phenotypes showed an over-representation (FDR≤5%). We replicated three of these phenotypes; situs inversus, heterotaxia, and double outlet right ventricle, in the general population cohort (FDR≤5%). Our findings lead us to propose that handedness is a polygenic trait controlled in part by the molecular mechanisms that establish LR body asymmetry early in development. © 2013 Brandler et al.

Tér és teljesség - Fenntarthatóság a területhasználatban és az építési tevékenységben

A cikk a fenntarthatóság témakörét a területhasználatra és az építési tevékenységre fókuszálva járja körül. Noha kész receptekkel nem szolgál, természet, társadalom és gazdaság egészlátó (integrált) szemlélete alapján néhány radikális, normalitást célzó – főként ökológiai gazdaságtani – szempontot fogalmaz meg, amelyekkel egy fenntarthatóbb rendszert alakíthatunk ki. Mivel az írást a fenntarthatóság kérdésköre ihlette, a természeti környezettel kapcsolatos megfontolások a kidolgozottabbak, s csak esetlegesen (ám elválaszthatatlanul) kapcsolódnak hozzájuk a társadalmi vonatkozások. A tanulmányból mindenesetre – implicit módon – így is kitűnik, hogy a fenntarthatóság eszméje (amely elsősorban a természeti korlátokból fakad) nem mond ellent a társadalmi-emberi kiteljesedésnek, azaz a közjónak, ha az embert és a társadalmat a maga teljességében szemléljük, s nem valamilyen torz és leegyszerűsítő (például a homo oeconomicus) ideológia alapján. A cikkben a radikális javaslatokat kevésbé radikális – környezetgazdasági – nézetek ellenpontozzák, mintegy érzékeltetve a problémakör és a megközelítésmódok rendkívüli összetettségét.

The moral economic man

Economic behavior is multifaceted and context-dependent. However, the so-called Homo Oeconomicus model states that agents are perfectly rational, self-interest-maximizing beings. This model can be criticized on both empirical and normative grounds. Understanding economic behavior requires a more complex and dynamic framework. In the "I & We" paradigm developed by Amitai Etzioni, economic behavior is co-determined by utility calculations and moral considerations. Two major factors can explain the ethicality of economic behavior; namely, the moral character of the agents and the relative cost of ethical behavior. Economic agents are moral beings, but the ethical fabric of the economy determines which face of the Moral Economic Man predominates.

Future of Capitalism

The moral foundation of capitalism should be reconsidered. Modern capitalism is disembedded from the social and cultural norms of society and produced a deep financial, ecological and social crisis. Competitiveness is the prevailing ideology of today’s business and economic policy. Companies, regions, and national economies seek to improve their productivity and gain competitive advantage. But these efforts often produce negative effects on various stakeholders at home and abroad. Competitiveness involves self-interest and aggressivity and produces monetary results at the expense of nature, society and future generations The collaborative enterprise framework promotes a view in which economic agents care about others and themselves and aim to create values for all the participants in their business ecosystems. Their criterion of success is mutually satisfying relationships with the stakeholders. New results of positive psychology and the Homo reciprocans model of behavioral sciences support this approach. The economic teachings of world religions challenge the way capitalism is functioning, and their corresponding perspectives are worthy of consideration. They represent life-serving modes of economizing which can assure the livelihood of human communities and the sustainability of natural ecosystems. Ethics and the future of capitalism are strongly connected. If we want to sustain capitalism for a long time we have to create a less violent, more caring form of it.

Characterization of L-Serine Deaminase and its Potential Role in the Pathogenicity of Pseudomonas aeruginosa

All pathogens require high energetic influxes to counterattack the host immune system and without this energy bacterial infections are easily cleared. This study is an investigation into one highly bioenergetic pathway in Pseudomonas aeruginosa involving the amino acid L-serine and the enzyme L-serine deaminase (L-SD). P. aeruginosa is an opportunistic pathogen causing infections in patients with compromised immune systems as well as patients with cystic fibrosis. Recent evidence has linked L-SD directly to the pathogenicity of several organisms including but not limited to Campylobacter jejuni, Mycobacterium bovis, Streptococcus pyogenes, and Yersinia pestis. We hypothesized that P. aeruginosa L-SD is likely to be critical for its virulence. Genome sequence analysis revealed the presence of two L-SD homo logs encoded by sdaA and sdaB. We analyzed the ability of P. aeruginosa to utilize serine and the role of SdaA and SdaB in serine deamination by comparing mutant strains of sdaA (PAOsdaA) and sdaB (PAOsdaB) with their isogenic parent P. aeruginosa P AO 1. We demonstrated that P. aeruginosa is unable to use serine as a sole carbon source. However, serine utilization is enhanced in the presence of glycine and this glycine-dependent induction of L-SD activity requires the inducer serine. The amino acid leucine was shown to inhibit L-SD activity from both SdaA and SdaB and the net contribution to L-serine deamination by SdaA and SdaB was ascertained at 34% and 66 %, respectively. These results suggest that P. aeruginosa LSD is quite different from the characterized E. coli L-SD that is glycine-independent but leucine-dependent for activation. Growth mutants able to use serine as a sole carbon source were also isolated and in addition, suicide vectors were constructed which allow for selective mutation of the sdaA and sdaB genes on any P. aeruginosa strain of interest. Future studies with a double mutant will reveal the importance of these genes for pathogenicity.

Climate Change and Regions at Risk: A Look at Central America

Climate change has been a security issue for mankind since Homo sapiens first emerged on the planet, driving him to find new and better food, water, shelter, and basic resources for survival and the advancement of civilization. Only recently, however, has the rate of climate change coupled with man’s knowledge of his own role in that change accelerated, perhaps profoundly, changing the security paradigm. If we take a ―decades‖ look at the security issue, we see competition for natural resources giving way to Cold War ideological containment and deterrence, itself giving way to non-state terrorism and extremism. While we continue to defend against these threats, we are faced with even greater security challenges that inextricably tie economic, food and human security together and where the flash points may not provide clearly discernable causes, as they will be intrinsically tied to climate change. Several scientific reports have revealed that the modest development gains that can be realized by some regions could be reversed by climate change. This means that climate change is not just a long-term environmental threat as was widely believed, but an economic and developmental disaster that is unfolding. As such, addressing climate change has become central to the development and poverty reduction by the World Bank and other financial institutions. In Latin America, poorer countries and communities, such as those found in Central America, will suffer the hardest because of weaker resilience and greater reliance on climatesensitive sectors such as agriculture. The US should attempt to deliver capability to assist these states to deal with the effects of climate change.

AS MARCAS E O CONSUMIDOR: Um estudo dos discursos da publicidade na perspectiva da modificação dos estereótipos na representação da família.The

Os papeis de homens e mulheres na sociedade contemporânea têm sido reestruturados, assim como a própria noção das estruturas familiares se renova. Algumas marcas estão atentas a essas mudanças e, no contexto mercadológico, fica evidente que os consumidores são diversos, com novas demandas e expectativas pelo reconhecimento dessa diversidade. Esta dissertação discute a mudança de abordagem na representação da família, demonstrando suas mutações de papeis na publicidade, esta vista como um produto sociocultural. Para tanto foi desenvolvida uma pesquisa qualitativa subsidiada pela análise de discurso de linha francesa. Primeiramente, o estudo qualitativo se desenvolve a partir de uma abordagem bibliográfica e documental e uma parte empírica de análise de discurso de campanhas publicitárias dos últimos anos, selecionadas a partir de seu foco nas representações familiares contemporâneas, de marcas que as tratem com naturalidade e demonstrem aceitar essas transições, buscando-as através da análise de casos múltiplos de amostras intencionais os avanços quando se trata dos modelos familiares, elementos que se desviam de representação conservadora da família. Demonstrou-se que há uma tendência de algumas marcas reforçarem qual é o público que desejam atingir através de suas construções discursivas que apontam para o reconhecimento das famílias reconstituídas, para a adoção e para a homo e monoparentalidade, ou seja, para a confirmação da existência dos variados tipos de família.

Caractérisation de l’activité Nur77 et de ses complexes en essais BRET par complémentation

Dans le système nerveux central, la dopamine joue un rôle crucial dans de nombreuses fonctions physiologiques telles que : l’apprentissage, le mouvement volontaire, la motivation, la cognition et la production hormonale. Il a été aussi démontré que le système de signalisation dopaminergique est altéré dans plusieurs maladies neurologiques et psychiatriques comme la maladie de Parkinson et la schizophrénie. Des études, effectuées dans le laboratoire du Dr.Daniel Lévesque (laboratoire d’accueil), ont montré que les récepteurs nucléaires Nur77 (NR4A1, NGFI-B) et RXRγ (retinoid X receptors γ) sont impliqués dans la régulation des effets de la dopamine dans le système nerveux central. De plus, ces données suggèrent que le complexe Nur77 et RXR joueraient un rôle crucial dans l’effet des médicaments antipsychotiques et antiparkinsoniens. Toutefois, très peu de médicaments ciblant Nur77 ont été identifiés à ce jour et les médicaments agissant sur RXRγ restent mal caractérisés. En outre, les analyses actuellement disponibles ne peuvent pas résumer la complexité des activités des NRs et génèrent des mesures indirectes des activités des drogues. Afin de mieux comprendre comment est régulée l’interaction Nur77/RXRγ dans ces processus, mon projet a été de mettre au point un essai BRET (Bioluminescence Resonance Energy Transfer) et PCA-BRET (Protein Complementation Assay-BRET) basé sur le recrutement d'un motif mimant un co-activateur fusionné avec la YFP. Nos différents essais ont été validés par courbes dose-réponse en utilisant différents composés RXR . Les EC50 (concentration efficace médiane, qui permet de mesurer l'efficacité d'un composé) obtenues étaient très semblables aux valeurs précédemment rapportées dans la littérature. Nous avons aussi pu identifier un composé le SR11237 (BMS649) qui semble posséder une sélectivité pour le complexe Nur77/RXRγ par rapport aux complexes Nurr1/RXRγ et RXRγ /RXRγ. Nos résultats indiquent que ces essais de BRET peuvent être utilisés pour évaluer la sélectivité de nouveaux composés pour les complexes Nur77/RXRγ, Nurr1/RXRγ et RXRγ /RXRγ. Un autre aspect de mon projet de doctorat a été de mettre en évidence par BRET l’importance de la SUMOylation dans la régulation de l'activité de Nur77 dans sa forme monomèrique, homodimèrique et hétérodimèrique. Nous avons ainsi identifié que Nur77 recrute principalement SUMO2 sur sa lysine 577. Il est intéressant de noté que le recrutement de la SUMO2 à Nur77 est potentialisé en présence de la SUMO E3 Ligase PIASγ. Aussi, la perte de la SUMOylation sur la lysine 577 entraîne l'incapacité de Nur77 de recruter divers motifs de co-activation mais pas pour ses formes homo- et hétérodimèrique. Cependant, la présence de PIASγ ne potentialise pas le recrutement du co-activateur, suggérant que cette SUMO E3 Ligase est seulement impliqué dans le processus de recrutement de la SUMO mais pas dans celui du co-activateur. Nous avons ainsi déterminé une nouvelle modification post-traductionnelle sur Nur77 régulant spécifiquement son activité monomérique Ces projets pourraient donc apporter de nouvelles données cruciales pour l’amélioration du traitement de la maladie de Parkinson ou de la schizophrénie, ainsi que d'obtenir une meilleure compréhension sur les mécanismes permettant la régulation de la fonction de Nur77

The role of the homeobox transcription factor Pitx3 in the mesencephalic dopaminergic system

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

PCR-Based Analysis of Mitochondrial DNA Copy Number, Mitochondrial DNA Damage, and Nuclear DNA Damage.

Because of the role that DNA damage and depletion play in human disease, it is important to develop and improve tools to assess these endpoints. This unit describes PCR-based methods to measure nuclear and mitochondrial DNA damage and copy number. Long amplicon quantitative polymerase chain reaction (LA-QPCR) is used to detect DNA damage by measuring the number of polymerase-inhibiting lesions present based on the amount of PCR amplification; real-time PCR (RT-PCR) is used to calculate genome content. In this unit, we provide step-by-step instructions to perform these assays in Homo sapiens, Mus musculus, Rattus norvegicus, Caenorhabditis elegans, Drosophila melanogaster, Danio rerio, Oryzias latipes, Fundulus grandis, and Fundulus heteroclitus, and discuss the advantages and disadvantages of these assays.

Ground and Electronic Excited States from Pairing Matrix Fluctuation and Particle-Particle Random Phase Approximation

The accurate description of ground and electronic excited states is an important and challenging topic in quantum chemistry. The pairing matrix fluctuation, as a counterpart of the density fluctuation, is applied to this topic. From the pairing matrix fluctuation, the exact electron correlation energy as well as two electron addition/removal energies can be extracted. Therefore, both ground state and excited states energies can be obtained and they are in principle exact with a complete knowledge of the pairing matrix fluctuation. In practice, considering the exact pairing matrix fluctuation is unknown, we adopt its simple approximation --- the particle-particle random phase approximation (pp-RPA) --- for ground and excited states calculations. The algorithms for accelerating the pp-RPA calculation, including spin separation, spin adaptation, as well as an iterative Davidson method, are developed. For ground states correlation descriptions, the results obtained from pp-RPA are usually comparable to and can be more accurate than those from traditional particle-hole random phase approximation (ph-RPA). For excited states, the pp-RPA is able to describe double, Rydberg, and charge transfer excitations, which are challenging for conventional time-dependent density functional theory (TDDFT). Although the pp-RPA intrinsically cannot describe those excitations excited from the orbitals below the highest occupied molecular orbital (HOMO), its performances on those single excitations that can be captured are comparable to TDDFT. The pp-RPA for excitation calculation is further applied to challenging diradical problems and is used to unveil the nature of the ground and electronic excited states of higher acenes. The pp-RPA and the corresponding Tamm-Dancoff approximation (pp-TDA) are also applied to conical intersections, an important concept in nonadiabatic dynamics. Their good description of the double-cone feature of conical intersections is in sharp contrast to the failure of TDDFT. All in all, the pairing matrix fluctuation opens up new channel of thinking for quantum chemistry, and the pp-RPA is a promising method in describing ground and electronic excited states.