Advantages such as chemical, mechanical and thermal stability together with high selectivity for the templated analyte render molecularly imprinted polymers MIPs interesting alternatives to routinely applied separation materials or antibodies. Nevertheless, many factors such as the choice of functional monomer, cross-linker, and porogenic solvent, as well as the ratio between template, functional monomer, and cross-linker will affect the resulting imprinting efficiency and polymer particle size and morphology. The research described in this thesis contributes to the development of new synthetic strategies for the generation of imprinted micro- and nanospheres for 17beta-estradiol (E2) focusing on accurate control and optimization of the governing parameters for precipitation polymerization, including the polymerization temperature and the cross-linker, yielding a one-step synthetic approach with superior control on the bead diameter, shape, monodispersity and imprinting efficiency. Thus synthesized imprinting materials for E2 were successfully applied in HPLC separation, solid phase extraction and radioligand binding assays. As the optimization of imprinted materials is based on fundamental understanding of the binding site properties, the investigations is aimed at establishing a more rational basis for further tailoring imprinted materials to the desired analytical application. The relationships between the particle porosity and rebinding properties were detailed, providing useful guidelines for controlling the particle properties for the desired application including, SPE pre-concentration, HPLC separations, and biomimetic binding assays. Furthermore, analytical techniques (1H-NMR and IR, etc.) and molecular modeling were combined in this thesis to facilitate advanced understanding of the fundamental principles governing selective recognition of molecularly imprinted polymers at a molecular level. The molecular interactions involved in the templating process of molecularly imprinted polymers based on the self-assembly approach were simulated in molecular dynamic simulation model by building a modeling system include all the imprinting components with correct ratio, which has never been reported before. Molecular level interactions such as hydrogen bonding, π-π stacking interactions as well as the free energy governing complex formation of E2 with the functional monomers 4-vinylpyridine (4VP) and methacrylic acid (MAA), and the cross-linker divinylbenzene (DVB) were discussed.

In this thesis I investigate whether human sexual preferences develop through sexual imprinting. Sexual imprinting is the acquisition of sexual preferences through non-rewarded experiences with parents and siblings during an early sensitive period and it is known to exist in many other animals. Learning is often sex specific so that males, for instance, learn to prefer as sexual partners individuals that look like their mother, and avoid individuals that look like their father. First, sexual imprinting in animals and humans is reviewed and compared to prevailing evolutionary views presupposing genetically determined sexual preferences. Further, by means of web surveys, I have explored the relationship between childhood exposure to parents with certain natural and cultural traits and sexual attraction to these traits in a partner. Cultural traits were included because it is unlikely that preferences for them are genetically determined adaptations. Parental effects varied between traits. For instance, in heterosexual males, a positive effect of mother was found on attraction to smoking, but not glasses, while a negative paternal effect was found on attraction to glasses, but not smoking. However, when maternal and paternal effects were investigated for a large number of artificial and natural traits, including smoking and glasses, an overall positive effect of opposite sex parent emerged in both heterosexual males and females. Additionally, in the last study we explored a sexual preference for pregnant and lactating women. Results suggest that exposure to a pregnant and lactating mother had an effect if it occurred when the respondent was between 1,5 and 5 years old. In conclusion, these results suggest that human sexual preferences are the result of sex specific learning during a sensitive period. Sexual imprinting should therefore be recognised as a plausible explanation to human sexual preferences that deserves further scientific investigation. / At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 3: Manuscript. Paper 4: Manuscript.

Made available in DSpace on 2014-06-11T19:35:11Z (GMT). No. of bitstreams: 0 Previous issue date: 2005-12-19Bitstream added on 2014-06-13T19:24:36Z : No. of bitstreams: 1 niciura_scm_dr_jabo.pdf: 1029270 bytes, checksum: 8a93cc4207fdb3426be68df4fa064ef9 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A maturação oocitária é marcada pela retomada da primeira divisão da meiose, com progressão do estádio de Vesícula Germinativa (GV) da Prófase I até a Metáfase II (MII), e inclui todos os eventos necessários para que o oócito expresse seu potencial máximo de desenvolvimento após a fecundação. Para avaliarmos a eficiência da maturação in vitro (MIV), utilizamos oócitos classificados em viáveis (graus I, II e III) e inviáveis (atrésico e desnudo), e acompanhamos a progressão nuclear e a distribuição dos grânulos corticais (GC) como indício de maturação citoplasmática, após MIV em TCM 199 com soro fetal bovino, hormônios, antibiótico e piruvato, por 24h em 5% de CO2 em ar. Maturação nuclear (78,4-87,8%) e citoplasmática (GC periféricos; 67,2-79,3%) foram semelhantes entre as diferentes classes de oócitos e apresentaramse como eventos independentes. Para o acompanhamento dos eventos desencadeados pelo espermatozóide, avaliamos a dinâmica nuclear e de microtúbulos, em intervalos de 2h, após fecundação in vitro (FIV), em meio TALP com heparina, PHE e sêmen preparado em gradiente de Percoll. Observamos que o estádio de MII foi predominante de 2 a 8h; MII e Anáfase/Telófase (A/T) predominaram às 10h; MII, A/T e estádio pronuclear (PN) de 14 a 16h; e PN a partir de 18h. A penetração do espermatozóide ocorreu após 4h da inseminação dos oócitos; a diferenciação dos PN 14 masculino e feminino pelo tamanho foi possível de 14 a 18h e a singamia ocorreu a partir de 24h. O período de 10h pode ser suficiente para que a FIV seja efetiva em oócitos bovinos, nas condições aqui descritas. / We aimed to evaluate events involved in in vitro maturation, fertilization and development, and parthenogenetic activation of bovine oocytes assessed by nuclear-cytoplasmic interaction and gene expression. Oocyte morphological selection did not affect nuclear maturation (78.4-87.8%) and cytoplasmic cortical granule distribution (67.2-79.3%). Following nuclear and microtubular dynamics after fertilization (IVF), we observed sperm penetration 4h after insemination; male and female pronuclei differentiation by size from 14 to 18h; syngamy after 24h; and sufficient co-incubation of spermatozoa and oocytes for 10h. Pronuclear transfer to study the interaction between nucleus (N) and cytoplasm (C) in parthenogenetic embryos produced by ionomycin followed by strontium (S) or 6-DMAP (D) was assessed by cleavage, eight-cell, and blastocyst development rates: CSND (76.5, 36.4, and 6.8%) and CDNS (69.5, 25.0, and 4.9%). S cytoplasm promoted dominant effect on D nucleus. Higher rates of developmental arrest up to the eight-cell stage were observed by the combination of cytoplasm and nucleus produced by the two different activation treatments. We recovered parthenogenetic D fetuses on Day 35, which were small but normal in formation and in appearance of chorio-alantoic membranes. Genomic imprinting of IGF2 was observed, but XIST was maternally expressed in extra-embryonic tissues. In vitro culture promoted higher expression of IGF2 and H19 genes and also increased IGF2/IGF2r ratio in IVF embryos compared to in vivo produced ones.

Orientador: Joaquim Mansano Garcia / Banca: Flávio Vieira Meirelles / Banca: Claudia Lima Verde Leal / Banca: Vera Fernanda Martins Hossepian de Lima / Banca: Gisele Zoccal Mingoti / Resumo: A maturação oocitária é marcada pela retomada da primeira divisão da meiose, com progressão do estádio de Vesícula Germinativa (GV) da Prófase I até a Metáfase II (MII), e inclui todos os eventos necessários para que o oócito expresse seu potencial máximo de desenvolvimento após a fecundação. Para avaliarmos a eficiência da maturação in vitro (MIV), utilizamos oócitos classificados em viáveis (graus I, II e III) e inviáveis (atrésico e desnudo), e acompanhamos a progressão nuclear e a distribuição dos grânulos corticais (GC) como indício de maturação citoplasmática, após MIV em TCM 199 com soro fetal bovino, hormônios, antibiótico e piruvato, por 24h em 5% de CO2 em ar. Maturação nuclear (78,4-87,8%) e citoplasmática (GC periféricos; 67,2-79,3%) foram semelhantes entre as diferentes classes de oócitos e apresentaramse como eventos independentes. Para o acompanhamento dos eventos desencadeados pelo espermatozóide, avaliamos a dinâmica nuclear e de microtúbulos, em intervalos de 2h, após fecundação in vitro (FIV), em meio TALP com heparina, PHE e sêmen preparado em gradiente de Percoll. Observamos que o estádio de MII foi predominante de 2 a 8h; MII e Anáfase/Telófase (A/T) predominaram às 10h; MII, A/T e estádio pronuclear (PN) de 14 a 16h; e PN a partir de 18h. A penetração do espermatozóide ocorreu após 4h da inseminação dos oócitos; a diferenciação dos PN 14 masculino e feminino pelo tamanho foi possível de 14 a 18h e a singamia ocorreu a partir de 24h. O período de 10h pode ser suficiente para que a FIV seja efetiva em oócitos bovinos, nas condições aqui descritas. / Abstract: We aimed to evaluate events involved in in vitro maturation, fertilization and development, and parthenogenetic activation of bovine oocytes assessed by nuclear-cytoplasmic interaction and gene expression. Oocyte morphological selection did not affect nuclear maturation (78.4-87.8%) and cytoplasmic cortical granule distribution (67.2-79.3%). Following nuclear and microtubular dynamics after fertilization (IVF), we observed sperm penetration 4h after insemination; male and female pronuclei differentiation by size from 14 to 18h; syngamy after 24h; and sufficient co-incubation of spermatozoa and oocytes for 10h. Pronuclear transfer to study the interaction between nucleus (N) and cytoplasm (C) in parthenogenetic embryos produced by ionomycin followed by strontium (S) or 6-DMAP (D) was assessed by cleavage, eight-cell, and blastocyst development rates: CSND (76.5, 36.4, and 6.8%) and CDNS (69.5, 25.0, and 4.9%). S cytoplasm promoted dominant effect on D nucleus. Higher rates of developmental arrest up to the eight-cell stage were observed by the combination of cytoplasm and nucleus produced by the two different activation treatments. We recovered parthenogenetic D fetuses on Day 35, which were small but normal in formation and in appearance of chorio-alantoic membranes. Genomic imprinting of IGF2 was observed, but XIST was maternally expressed in extra-embryonic tissues. In vitro culture promoted higher expression of IGF2 and H19 genes and also increased IGF2/IGF2r ratio in IVF embryos compared to in vivo produced ones. / Doutor

The syntheses and evaluation of oxidovanadium(IV) complexes as catalysts for the oxidation of refractory organosulfur compounds in fuels is presented. The sulfones produced from the oxidation reaction were removed from fuel oils by employing molecularly imprinted polymers (MIPs). The oxidovanadium(IV) homogeneous catalyst, [V ͥ ͮ O(sal-HBPD)], as well as its heterogeneous polymer supported derivatives, poly[V ͥ ͮ O(sal-AHBPD)] and poly[V ͥ ͮ O(allylSB-co-EGDMA)], were synthesized and fully characterized by elemental analysis, FTIR, UV-Vis, XPS, AFM, SEM, BET and single crystal XRD for [V ͥ ͮ O(sal-HBPD)]. The MIPs were also characterized by elemental analysis, FTIR, SEM, EDX and BET. The catalyzed oxidation of fuel oil model sulfur compounds, thiophene (TH), benzothiophene (BT), dibenzothiophene (DBT) and 4,6-dimethyldibenzothiophene (4,6-DMDBT), was conducted under batch and continuous flow processes at 40°C by using tert-butylhydroperoxide (t-BuOOH) as oxidant. The continuous flow oxidation process presented the highest overall conversions and very high selectivity for sulfones. Maximum oxidation conversions of 71%, 89%, 99% and 88% was achieved for TH, BT, DBT and 4,6-DMDBT respectively when poly[V ͥ ͮ O(allylSB-co-EGDMA)] was employed at a flow-rate of 1 mL/h with over 90% sulfone selectivity. The process was further applied to the oxidation of hydro-treated diesel containing 385 ± 4.6 ppm of sulfur (mainly dibenzothiophene and dibenzothiophene derivatives), and this resulted to a high sulfur oxidation yield (> 99%), thus producing polar sulfones which are extractible by polar solid phase extractants. Adsorption of the polar sulfone compounds was carried-out by employing MIPs which were fabricated through the formation of recognition sites complementary to oxidized sulfur-containing compounds (sulfones) on electrospun polybenzimidazole (PBI) nanofibers, cross-linked chitosan microspheres and electrospun chitosan nanofibers. Adsorption of benzothiophene sulfone (BTO₂), dibenzothiophene sulfone (DBTO₂) and 4,6-dimethyldibenzothiophene sulfone (4,6-DMDBTO₂) on the various molecularly imprinted adsorbents presented a Freundlich (multi-layered) adsorption isotherm which indicated interaction of adsorbed organosulfur compounds. Maximum adsorption observed for BTO₂, DBTO₂ and 4,6-DMDBTO₂ respectively was 8.5 ± 0.6 mg/g, 7.0 ± 0.5 mg/g and 6.6 ± 0.7 mg/g when imprinted chitosan nanofibers were employed, 4.9 ± 0.5 mg/g, 4.2 ± 0.7 mg/g and 3.9 ± 0.6 mg/g on molecularly imprinted chitosan microspheres, and 28.5 ± 0.4 mg/g, 29.8 ± 2.2 mg/g and 20.1 ± 1.4 mg/g on molecularly imprinted PBI nanofibers. Application of electrospun chitosan nanofibers on oxidized hydro-treated diesel presented a sulfur removal capacity of 84%, leaving 62 ± 3.2 ppm S in the fuel, while imprinted PBI electrospun nanofibers displayed excellent sulfur removal, keeping sulfur in the fuel after the oxidation/adsorption below the determined limit of detection (LOD), which is 2.4 ppm S. The high level of sulfur removal displayed by imprinted PBI nanofibers was ascribed to hydrogen bonding effects, and π-π stacking between aromatic sulfone compounds and the benzimidazole ring which were confirmed by chemical modelling with density functional theory (DFT) as well as the imprinting effect. The home-made pressurized hot water extraction (PHWE) system was applied for extraction/desorption of sulfone compounds adsorbed on the PBI nanofibers at a flow rate of 1 mL/min and at 150°C with an applied pressure of 30 bars. Application of molecularly imprinted PBI nanofibers for the desulfurization of oxidized hydro-treated fuel showed potential for use in refining industries to reach ultra-low sulfur fuel level, which falls below the 10 ppm sulfur limit which is mandated by the environmental protection agency (EPA) from 2015.

Pw1/Peg3 est un gène d’empreinte parental exprimé par l’allèle paternel. Il est exprimé dans l’ensemble des populations de cellules souches, y compris les cellules satellites du tissu musculaire. Nous avons découvert que la perte constitutive de Pw1/Peg3 entraîne une perte de la masse musculaire, résultat d’une diminution du nombre de fibres musculaires. Le nombre de fibres réduit est présent dès la naissance. De plus, les souris double KO ont un nombre de fibres encore inférieur, suggérant que l’allèle maternel est fonctionnel pendant le développement pré-natal, et des analyses de souris hybrides C57BL6J/CAST/Ei révèlent une expression bi-allélique de Pw1/Peg3 d’environ 10%. Pw1/Peg3 est également fortement exprimé après blessure du muscle squelettique. Chez les souris Pw1/Peg3 KO, nous avons observé que les cellules satellites montrent une réduction de leur capacité d’auto-renouvèlement à la suite d’une blessure. Pw1/Peg3 est également exprimé dans une sous-population de cellules souches interstitielles, les PICS. Afin de déterminer le rôle spécifique de Pw1/Peg3 dans les cellules satellites nous avons croisé notre allèle conditionnel Pw1/Peg3 avec la lignée Pax7-Cre-ER. Ces souris ont un phénotype présentant un défaut de régénération prononcé, montrant ainsi un rôle clair et direct de Pw1/Peg3 dans la fonction régénératrice des cellules satellites. En résumé, l’ensemble de ces données montre un rôle de Pw1/Peg3 dans le développement fœtal et la détermination du nombre de fibres musculaires par son action dans l’auto-renouvellement des cellules satellites du tissu musculaire. / Pw1/Peg3 is a parentally imprinted gene expressed from the paternal allele. It is expressed in all adult progenitor/stem cell populations examined to date including muscle satellite cells. We examined the impact of loss-of-function of Pw1/Peg3 in skeletal muscle, a tissue that greatly contributes to body mass. We found that constitutive loss of Pw1/Peg3 results in reduced muscle mass resulting from a decrease in muscle fiber number. The reduced fiber number is present at birth. Mice lacking both the paternal and maternal alleles display a lower fiber number as compared to mice carrying the paternal deletion, suggesting that the maternal allele is functional during prenatal development. Hybrid analyses (C57BL6J and Cast/Ei) of muscle tissue reveal a bi-allelic expression of Pw1/Peg3 around 10%. Pw1/Peg3 is strongly up-regulated in response to muscle injury. Using the constitutive Pw1/Peg3 knock out mouse, we observed that satellite cells display a reduced self-renewal capacity following muscle injury. Pw1/Peg3 is expressed in satellite cells as well as a subset of muscle interstitial cells (PICs). To determine the specific role of Pw1/Peg3 in satellite cells, we crossed our conditional Pw1/Peg3 allele with the Pax7-CreER line. Interestingly, these mice displayed a more pronounced phenotype of impaired regeneration revealing a clear and direct role for Pw1/Peg3 in satellite cells. Taken together, our data show that Pw1/Peg3 plays a role during fetal development in the determination of muscle fiber number that is gene-dosage dependent and plays a specific role in muscle satellite cell self-renewal.

Chez les mammifères, une centaine de gènes sont soumis à une régulation épigénétique où seule la copie maternelle, ou paternelle, est exprimée. Ce phénomène appelé empreinte parentale alimente encore différentes théories liées à la reproduction, notamment celles du conflit parental et de la coadaptation entre mère et enfant. Pw1/Peg3 est un gène d'empreinte paternellement exprimé. Cependant, à l'aide de deux modèles de souris bien distincts, une souris rapporteur (Pw1IRESnLacZ) et une nouvelle souris knockout pour Pw1/Peg3, nous avons détecté des transcrits Pw1/Peg3 maternels dans le cerveau périnatal. Plus précisément, nous avons mis en évidence une expression bi-allélique du gène rapporteur Pw1IRESnLacZ restreinte aux deux futures niches de cellules souches neurales adultes. In vitro, nous avons conclu, via des cultures primaires de cellules souches neurales, que l'expression bi-allélique endogène de Pw1/Peg3 est un évènement ponctuel rare. D'ailleurs lors de la caractérisation de notre modèle de souris Pw1/Peg3 knockout, nous avons observé un retard de croissance uniquement lors de la délétion de l'allèle Pw1/Peg3 paternel. Ce phénotype n'est pas lié à un problème de prise alimentaire chez les nouveaux-nés et contrairement à ce qui a été précédemment décrit, nous n'avons détecté aucun défaut de comportement maternel chez les femelles mutantes pour Pw1/Peg3. La lactation n'est pas non plus impactée par la délétion de Pw1/Peg3. Ces résultats démontrent que Pw1/Peg3 favorise intrinsèquement la croissance postnatale et que, désormais, ce gène d'empreinte ne peut plus être utilisé afin d'illustrer la théorie de coadaptation entre mère et enfant. / In mammals, a hundred of genes are preferentially expressed from one specific parental allele; a phenomenon referred as genomic imprinting. Establishing theories to explain the emergence of such a gene dosage strategy is challenging. Pw1/Peg3 is a paternally expressed gene. Using both a reporter mouse model and a novel constitutive knockout mouse model, we detected Pw1/Peg3 transcription from the maternal allele, which is normally silent, in the perinatal brain. Specifically, we observed that a putative Pw1/Peg3 bi-allelic expression is mainly restricted to the two future adult neural stem cells niches. In vitro experiments on primary neural stem cells allowed us to conclude that imprinting relaxation of the Pw1/Peg3 maternal allele is a rare event. Whether it affects the mouse phenotype is currently under investigation. In parallel, consistent with previously established mutant mouse models we confirmed that paternal Pw1/Peg3 deletion leads to growth retardation. However we did not find any impairment in maternal behaviors upon heterozygous or homozygous loss of Pw1/Peg3. Lactation was also not disrupted and mutant pups exhibited a normal suckling ability. Taken together, PW1/PEG3 promotes growth intrinsically and can no longer be used to illustrate the popular coadaptation theory between mother and infant.

Previous studies have shown that women choose partners resembling their fathers in various characteristics. However, none of the studies have focused on woman's father-partner body odour similarity, even though body odour plays an important role in mate choice. The main aim of our study was to test whether the woman's father and partner body odour is similar and whether the rating of this similarity was affected by the body odour's intensity and pleasantness. Further, we wanted to know whether the quality of woman's relationship with her father during her childhood influences the body odour similarity. We also tested the effect of woman's father-partner body odour similarity on sexual and relationship satisfaction. Twenty-five women with their fathers and partners participated in the study. Every respondent completed a set of questionnaires, in addition, fathers and partners provided samples of their body odours. Body odour similarity was rated by independent female raters (N=128). According to the results of our study, body odour of woman's father and partner is significantly similar. The body odour was also similar in intensity and pleasantness. The quality of woman's relationship with her father does not affect woman's father-partner body odour similarity. Nevertheless, this similarity...

<p>Laser shock imprinting (LSI) is a novel fabrication technique capable of manufacturing various membrane materials. This top-down imprinting process can fabricate membranes in high precision, high throughput, and large scalability. It reveals a variety of applications ranging from electronics to photonics, which is beneficial from its reliable and precise modulation of micro/nanostructures. </p> <p> In this thesis, we firstly proposed and developed a cost-effective LSI process to manufacture hierarchical micro/nanostructured power generators. By combining the conventional soft lithography technique, LSI is well compatible with it to fabricate metal membranes towards soft templates. It is a significant progress from the originally-developed silicon wafer template layout because it effectively reduces the process cost by replacing sophisticatedly developed silicon wafers with low-cost photocurable polymers. In addition, the use of polymer expands the boundary limit of geometrical complexity from simple patterns to hierarchical structures, as a result, we successfully conducted LSI technology to fabricate biomimic leaf structures into metallic membranes with the help of soft SU-8 templates. These fabricated metallic membraned are used as water-driven triboelectric nanogenerators. In addition to the introduction of polymer template, we further developed a successive laser shock imprinting (SLSI) process to fabricate hierarchical nanostructures in a higher resolution. Typically, grating templates are collected via recycling blank discs and used as soft templates. Then multiple times of LSI process are conducted to manufacture membranes into complex nanostructures. The use of blank disc further reduces cost and increase process resolution. The highlight of this part of work is to feature the introduction of metallic thin films on disc template, which plays a significant role during this high strain rate imprinting process. Then, the imprinting mechanism was investigated through the finite element method to validate the experimental findings. Lastly, this soft template LSI process was applied to fabricate low dimensional materials such as nanowires (1D) and nanomembranes (2D), potentially introducing homogeneous and inhomogeneous strain field. Kelvin probe force microscopy was used to directly probe strain-induced changes. This soft-template LSI process reveals a new route of precisely fabricating low dimensional membranes into nanoelectronics systems. </p>

24 pairs of newly-hatched Leghorn chicks were randomly divided into 3 groups. Ss in one group followed a moving object for 30 min on Day 1 of life; Ss in another group followed for 30 min on Day 2; Ss in the remaining group followed for 15 min on Day 1 and 15 min on Day 2. One S in each pair followed by his own effort, while the other S rode behind the object in a transparent box. On Day 4, Ss were tested for the duration of following of the object. No important differences among groups were observed. On Day 6, Ss were tested for ability to discriminate between the original and a novel object, and for following the original. Active Ss scored significantly higher than passive Ss on all Day 6 tests; Ss trained on Day 2 scored significantly higher on the following than Ss trained on Day 1. The results suggest that the ‘law of effort’ may apply more to discrimination than to recognition of the imprinting object.