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Heterosexual romance and marriage are institutionalized ideals in our society, set forth, in part, through the portrayal of stereotyped gender roles in fairy tales, such as Cinderella, and by the mainstream media. This thesis explores the cultural messages aimed at women, which impose the necessity of altering oneself to achieve marriage, and offers feminist viewpoints. Using the form of the personal essay, I discuss the ideals of Cinderella, Prince Charming, marriage, and Happily Ever After as unrealistic, though still prevalent, given the popularity of books like The Rules: Time-tested Secrets for Capturing the Heart of Mr. Right, and Princess Diana as Cinderella icon. Essays on my own experience of marriage and divorce supplement the cultural issues, juxtaposing the personal and political toward a new paradigm for relationships.

NorM multidrug transport proteins belong to the multiple antibiotics and toxins extrusion (MATE) family of secondary active transporters. Members of this family are present across all species including bacteria, plants and humans. In bacteria, their over-expression can lead to antibiotic resistance, whereas in the human body, the transporters can alter the plasma levels of drugs. NorM proteins are therefore relevant for the pharmacokinetic properties of drugs. Previously, NorM from Vibrio cholerae (NorM-VC) was shown to export drug (ethidium) in an antiport reaction that is coupled to the simultaneous uptake of protons and sodium ions down their electrochemical gradients across the plasma membrane. But NorM from Pseudomonas stutzeri (NorM-PS) was shown to transport DAPI by utilising proton cycling exclusively. NorM-VC and NorM-PS share 42% identical amino-acid residues and yet their functions differ in terms of their ion coupling properties. These differences in functionality of two highly homologous proteins provide an excellent opportunity to carry out a comparative study. The work presented in this thesis investigates the energetics of drug transport processes by NorM-VC and NorM-PS and the structural basis for ion-coupled drug transport by NorM-VC. Ethidium efflux assays in intact Lactococcus lactis cells were used to study the effect of the magnitude and composition of the proton- and sodium-motive force on transport activity. Furthermore, ethidium binding assays were used to study partial reactions in drug efflux processes. These biochemical data were supplemented by computational studies and analyses of current protein structures. Based on the observations detailed here, a novel transport model for NorM-VC is proposed, which explains published findings for NorM-VC and other MATE transporters. The model represents a potentially universal mechanism for MATE transporters that can be used to predict further structure-function relationships in this important family of member transporters.

Orientador: Priscila Alessio Constantino / Resumo: O presente trabalho descreve o estudo da interação entre o lipídio AA (ácido araquídico), os fosfolipídios DPPC (1,2-dipalmitoil-sn-glicero-3-fosfatidilcolina) e DPPG (1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol) com a enzima Ty (tirosinase). Através da interação monocamada lipídica/enzima foi desenvolvido um biossensor para a detecção de polifenóis. O estudo de interação foi realizado via filmes de Langmuir, ou seja, monocamadas lipídicas dispostas na interface ar/água que mimetizaram modelos de membranas biológicas, sendo estes analisados através de isotermas (π-A) e do módulo de compressibilidade. Além disso, foram realizadas medidas de PM-IRRAS (do inglês - Polarization Modulation - Infrared Reflection-Adsorption Spectroscopy) que auxiliaram no estudo da interação membrana/enzima em nível molecular. Por meio desses estudos foi possível observar uma maior interação e incorporação da enzima nas monocamadas de DPPC/Ty. Então, os filmes de Langmuir foram depositados em ITO (substrato eletrocondutor de vidro recoberto com óxido de estanho dopado com índio) utilizando a técnica de Langmuir-Blodgett (LB). Os filmes foram avaliados como biossensores para detecção de polifenóis em amostras padrões de catecol por voltametria cíclica. Nessa etapa, foram avaliados os efeitos das matrizes lipídicas, número de camadas e concentração do eletrólito de suporte no processo eletrocatalítico de redução do catecol. Além disso, foi avaliado o efeito da incorporação do mediador eletrônico LuPc... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: This work describes the study of interaction between lipid AA (arachidic acid), phospholipids DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine) and DPPG (1,2-dipalmitoyl-snglycero-3-phosphoglycerol) with the enzyme Ty (tyrosinase). A biosensor for the detection of polyphenols was developed through the interaction of lipid monolayer/enzyme. The study of interaction was performed in Langmuir films, in other words, lipids monolayer organized at the air/water interface simulated biological membrane models, using the isotherms (π-A) and the compressibility module. In addition, measurements of PM-IRRAS (Polarization Modulation - Infrared Reflection-adsorption Spectroscopy) were performed to study of membrane/enzyme interaction at the molecular level. In these studies, it was possible to observe a strong interaction and incorporation of the enzyme in DPPC monolayer. Thus, the films were grown on ITO (indium doped tin oxide glass substrate) using the Langmuir-Blodgett (LB) technique. The films were evaluated as biosensors for detection of polyphenols in standard catechol samples using cyclic voltammetry. At this stage, the effects of lipid matrices, number of layers and support electrolyte concentration on the electrocatalytic catechol reduction process were evaluated. After that, the effect of the incorporation of the electron mediator LuPc2 (Lutetium bisphthalocyanine) into the lipid matrix was evaluated. Electrochemical studies indicated that DPPC/Ty and DPPC:LuPc2/Ty films,... (Complete abstract click electronic access below) / Mestre

The release of the neurotoxic alkaloid saxitoxin by cyanobacterial cells was previously thought to occur primarily after cell lysis, yet recent evidence also suggests active toxin export by membrane transporters. Transporter proteins associated with STX biosynthesis in Cylindrospermopsis raciborskii T3 (sxtF and sxtM) and Anabaena circinalis 131C (naDt) were predicted to be involved in the export of STX from cyanobacterial cells. The main aim of this project was to characterise the transporters associated with STX biosynthesis, by investigation of their genetic prevalence, functional substrates and specific regulation. An sxtM homologue was discovered in A. circinalis 131C, as part of an sxt cluster, and found to be uniquely associated with STX-producing strains. Bioinformatic and phylogenetic analysis showed that the translated sxt transporters clustered with the NorM prokaryotic MATE sub-family and membrane topology analysis predicted 12 membrane-spanning regions. To characterise the functional substrates of the putative STX-transporters, they were heterologously expressed in the antibiotic-sensitive E. coli strain KAM32. Expression of the sxt MATES complemented host sensitivity to the cationic fluroquinolone antibiotics, ciprofloxacin and ofloxacin. Disruption of gene homologues of naDt and the sxt MATE genes in Synechocystis sp. PCC6803 yielded mutant strains with increased sensitivity to the toxic organic cations, methyl viologen and acriflavine. Transcription of the putative STX transporters, and the putative STX biosynthesis gene sxtA, was studied in C. raciborskii T3 and A. circinalis 131C under alkali and Na+ stress. Alkali stress (pH 9) decreased total STX levels in A. circinalis 131C and was correlated with a down-regulation of the putative transport and biosynthetic genes. In C. raciborskii T3, alkali stress promoted higher extracellular but lower intracellular STX levels, which also correlated with large increases in transcription of the putative STX transport genes.

The evolution of mate choice remains one of the most controversial topics within evolutionary biology. In particular, the coevolutionary dynamics between ornaments and mating preferences has been extensively studied, but few generalizations have emerged. From a theoretical standpoint, the nature of the genetic covariance built up by the process of mate choice has received considerable attention, though the models still make biologically unrealistic assumptions. Empirically, the difficulty of estimating parameters in the models has hindered our ability to understand what processes are occurring in nature. Thus, it is the goal of this dissertation to contribute to the field both theoretically and empirically. I begin with a review of the evolution of mate choice and demonstrate how the lack of cross-talk between theoretical and empirical pursuits into studying mate choice has constrained our ability to extract basic principles. The review is followed by a new model of intersexual selection that relaxes some of the critical assumptions inherent in sexual selection theory. There are two empirical studies whose goal is to measure mating preference functions and genetic correlations in a way that can be related back to theory. Finally, I conclude by setting the stage for future endeavors into exploring the evolution of mate choice. The results presented herein demonstrate four things: (i) a lack of communication between theoretical and empirical studies of mate choice; (ii) genetic drift plays a much larger role in preference evolution than previously demonstrated; (iii) genetic correlations other than those explicitly modeled are likely to be important in preference evolution; and (iv) variation in mating preferences can eliminate intersexual selection altogether. From these four findings it can be concluded that a tighter link between theory and empiricism is needed, with a particular emphasis on the importance of measuring individual-level preference functions. Models will benefit from integrating the specific phenotypes measured by empiricists. Experimentation will be more useful to theory if particular attention is paid to the exact phenotypes that are measured. Overall, this dissertation is a stepping stone for a more cohesive and accurate understanding of mate choice evolution.

My Ph.D. research has examined the evolution of mating preferences and their role in speciation. I have addressed these topics empirically, using sympatric species pairs of limnetic and benthic threespine sticklebacks, and theoretically, with multilocus population genetics. Sticklebacks are small fish that occur in lakes, streams and estuaries throughout British Columbia. Most lakes contain one type of stickleback, however, several lakes support two differentially adapted sympatric species: a large benthic form (benthic), and a smaller zooplanktivorous form (limnetic). Chapter 2 examines the role of species interactions in shaping male mating preferences. I determined that the mate preferences of the small species have shifted from preferring large females (the ancestral state) to preferring small females due either to selection against hybrids (reinforcement), or to egg predation by the larger benthic species. Chapter 3 explores the idea that sexual imprinting may facilitate assortative mating between benthic and limnetic sticklebacks. Sexual imprinting occurs when individuals imprint on the phenotype of their parents, and subsequently prefer mates that resemble their parents. The results suggested that sexual imprinting does not contribute to assortative mating between the sympatric species pairs, implying that genetics are more important than early learning for the formation of mate preferences. Chapter 4 focuses on differences in male breeding colour between benthics and limnetics. The results reveal that limnetic males have more intense red and blue coloration than benthic and solitary males. These differences in colour could be due to reinforcement, to differences in visual sensitivity of females, or to territorial interactions between males. Chapter 5 examines the evolution of female mating preferences under different scenarios of sex-linkage, when the male display trait is sexually antagonistic. Theoretical analysis suggests that sexually antagonistic traits on the X chromosome (males XY, females XX), females will evolve to prefer mates carrying alleles beneficial to their daughters. In contrast, with a Z-linked trait (males ZZ, females ZW), females more often evolve preferences for mates carrying alleles beneficial to their sons (e.g., flashy displays). This provides an explanation for why males in ZW species have more elaborate sexual displays than males in XY species.

My Ph.D. research has examined the evolution of mating preferences and their role in speciation. I have addressed these topics empirically, using sympatric species pairs of limnetic and benthic threespine sticklebacks, and theoretically, with multilocus population genetics. Sticklebacks are small fish that occur in lakes, streams and estuaries throughout British Columbia. Most lakes contain one type of stickleback, however, several lakes support two differentially adapted sympatric species: a large benthic form (benthic), and a smaller zooplanktivorous form (limnetic). Chapter 2 examines the role of species interactions in shaping male mating preferences. I determined that the mate preferences of the small species have shifted from preferring large females (the ancestral state) to preferring small females due either to selection against hybrids (reinforcement), or to egg predation by the larger benthic species. Chapter 3 explores the idea that sexual imprinting may facilitate assortative mating between benthic and limnetic sticklebacks. Sexual imprinting occurs when individuals imprint on the phenotype of their parents, and subsequently prefer mates that resemble their parents. The results suggested that sexual imprinting does not contribute to assortative mating between the sympatric species pairs, implying that genetics are more important than early learning for the formation of mate preferences. Chapter 4 focuses on differences in male breeding colour between benthics and limnetics. The results reveal that limnetic males have more intense red and blue coloration than benthic and solitary males. These differences in colour could be due to reinforcement, to differences in visual sensitivity of females, or to territorial interactions between males. Chapter 5 examines the evolution of female mating preferences under different scenarios of sex-linkage, when the male display trait is sexually antagonistic. Theoretical analysis suggests that sexually antagonistic traits on the X chromosome (males XY, females XX), females will evolve to prefer mates carrying alleles beneficial to their daughters. In contrast, with a Z-linked trait (males ZZ, females ZW), females more often evolve preferences for mates carrying alleles beneficial to their sons (e.g., flashy displays). This provides an explanation for why males in ZW species have more elaborate sexual displays than males in XY species.

Thesis (M.A.)--York University, 1998. Graduate Programme in Psychology. / Typescript. Includes bibliographical references (leaves 49-51). Also available on the Internet. MODE OF ACCESS via web browser by entering the following URL:http://wwwlib.umi.com/cr/yorku/fullcit?pMQ27383.

The ‘good genes’ model of sexual selection predicts that sexual and natural selection should act concordantly. However sexual selection can favour alleles in males that are costly when expressed in daughters, placing the two in opposition. The relationship between natural and sexual selection depends on the nature of genetic variation for fitness. Laboratory adaptation may deplete sexually concordant fitness variation, overestimating sexually antagonistic variation and obscuring good genes. I investigated sire-offspring fitness correlations in Drosophila melanogaster populations expected to differ in their levels of sexually concordant fitness variation. In maladapted populations, successful sires produced fitter daughters than unsuccessful sires; this pattern was reversed in adapted populations. Several generations later, successful sires in both population types produced lower fitness daughters than unsuccessful sires, consistent with predictions. However, subsequent generations revealed no effect of sire status on daughter fitness, highlighting the difficulty in testing predictions on the evolutionary dynamics of fitness heritability.