The research presented in this dissertation involves the development of chiral and achiral analysis using capillary electrochromatography (CEC) and CEC coupled to mass spectrometry (CEC-MS). Chapter 1 briefly reviews CEC fundamentals and latest development on chiral CEC and CEC-MS. The CEC-UV enantioseparations for several acidic compounds are described in Chapter 2. The optimum resolutions for these acidic enantiomers are achieved in ion-suppression mode, i.e. with an acidic mobile phase. One of major drawback in coupling CEC with MS is the bubble formation at the column outlet end, resulting in irreproducible retention time and erratic baseline, or even current breakdown. By introducing internal tapered columns, the aforementioned limitations of CEC-MS are successfully overcome in Chapter 3. The CEC-MS enantioseparation of warfarin and coumachlor is carefully investigated and applied to quantify R- and S-warfarin in human plasma. For individual enantiomers, a concentration of 25 ng/mL is detectable. To further improve the robustness of CEC-MS column, a new procedure of fabricating internal tapered columns is reported in Chapter 4. These internal tapered columns demonstrate excellent ruggedness, low background noise, and good compatibility in reversed-phase and polar organic modes of CEC-MS. In chapter 5, the feasibility of using internal tapered columns packed with vancomycin chiral stationary phase (CSP) is explored for simultaneous enantioseparation of eight â-blockers using CEC-MS. After a careful optimization of the mobile phase composition, sheath liquid and spray chamber parameter, 15 out of 16 enantiomers could be simultaneously resolved with excellent efficiency and detection sensitivity. The synthesis and characterization of sulfated and sulfonated cellulose phenylcarbamate CSPs is described in Chapter 6. The use of these CSPs, especially the sulfonated one, significantly enhances the EOF profile and sample throughput but maintain high enantiomeric resolving power under various modes of CEC and CEC-MS. By combining CEC and atmospheric pressure photo-ionization (APPI) MS, Chapter 7 demonstrates the separation and detection of mono-methylated benzo[a]pyrene (MBAP) isomers with ~100 times enhancement on detection sensitivity than CEC-UV. In Appedix 2, monolithic columns are synthesized through photopolymerized sol-gel approach and utilized for CEC and CEC-APPI-MS of polyaromatic hydrocarbons, and alkyl phenyl ketones.

Le développement de nouvelles molécules capables d’intéragir avec l’ADN, leur mode d’intéraction et leur affinité est un domaine de recherche particulièrement important. Dans ce travail nous avons étudié les interactions avec l’ADN de la molécule Hoechst 33258 connu pour être un ligand du petit sillon ainsi que plusieurs de ces analogues.Dans un premier temps nous avons étudié la stabilité des complexes ADN-Hoechst 33258 en solution avec et sans DMSO comme co-solvant. Deuxièmement, les affinités de dérivés nouvellement conçus et synthétisés du Hoechst 33258 vis-à-vis de l'ADN ont été évaluées. Enfin, nous avons étudiés la capacité d’induction d’excimers en présence d’ADN de différentes molécules pyrénylées. Ces études ont été effectuées par différentes méthodes spectroscopiques, telles que l'absorbance UV-visible, la fluorescence, le dichroïsme circulaire, la spectroscopie de masse ESI et de la modélisation moléculaire. / The development of new DNA binders and the evaluation of their affinity toward DNA as well as their mode of binding is an area of research of prime importance. In this thesis we studied the interactions of Hoechst 33258, a well-known groove binder, as well as some of its newly synthesized derivatives with DNA. The stability of DNA-Hoechst 33258 complex in solution with and without DMSO as a co-solvent was evaluated.Secondly, the affinities of newly designed and synthesized derivatives of Hoechst 33258 toward DNA were evaluated. Finally, a set of pyrene derivatives able to induced excimer formation upon binding to DNA were studied. Different spectroscopic methods, such as UV-vis absorbance, fluorescence, circular dichroism, ESI mass spectroscopy and molecular docking were applied for the complete evaluation of the affinity of these ligands toward DNA.

Made available in DSpace on 2015-04-11T13:55:53Z (GMT). No. of bitstreams: 1 joelma keith.pdf: 1871679 bytes, checksum: beec1d97b696ab3341d4cc1afa60a8ad (MD5) Previous issue date: 2013-07-29 / Fundação de Amparo à Pesquisa do Estado do Amazonas / Bactris maraja Mart ., commonly known as maraja, is a palm native to the Amazon use little known. However, the trunks are used by traditional construction of corrals fishing. The fruits are edible, being consumed by humans, and especially the wildlife. Studies with palm trees in the areas of technology and morphoanatomic have been conducted, however, the peculiarities inherent to the species of the family Arecaceae, difficult material handling. This research aimed to examine seeds of B. maraja regarding the structure and physiological quality, as well as describe the morphology development and germination plantular, beyond morphoanatomy seedling. The clusters containing mature fruits were collected from 25 individuals in floodplain environment in the municipality of Careiro Lowland, Amazonas. Pyrenes and the seeds had a water content of 23,1 % and 17,9 %, respectively. The pyrene is oval to elliptical, with core disk formed by sclerified cells thick-walled, however, absent in the region of the germ pore. The seed is eurispermica, being cordate, oblong and elliptical. Shows two to three pores. The integument is thin, consisting of sclerenchyma cells and thickened with phenolic content. The endosperm is homogeneous, white and rigid. The embryo is apical, small, tapered and milky-white, with-embryonic axis oblique and curved, with three leaf primordia. The endosperm storage protein and lipid reserves. The alternating temperature of 26-40 °C was suitable for seed germination, regardless of the level of processing. The progressive drying of pyrene facilitated the extraction of seeds. However, drying and removal of the endocarp reduced germination and vigor. Therefore, the processing is not recommended. The seed soaking for two days enhanced germination. The seed dormancy is not caused by the presence of the endocarp. Germination is adjacent ligulate, cryptocotylar and hypogeal. Started at 152 days after sowing, with protrusion of the cotyledon petiole, forming the germinal button to 160 days. The seedling completed the development to 273 with the depletion of the haustorium. The eophyll is bifid, with tetracytical stomata, multicellular trichomes and collateral vascular bundles. The root shows thick epidermis, cortex sclerenchymatous and parenchymatous, endoderm conspicuous; polyarc is with hollow vascular cylinder with spinal sclerenchymatic. / Bactris maraja Mart., conhecida vulgarmente como marajá, é uma palmeira nativa da Amazônia de uso pouco conhecido. Contudo, os estipes são utilizados pelas populações tradicionais na construção de currais de pesca artesanal. Os frutos são comestíveis, sendo consumidos pelo ser humano e, sobretudo, pela fauna silvestre. Estudos com palmeiras nas áreas tecnológicas e morfoanatômicas têm sido realizados, contudo, peculiaridades intrínsecas às espécies da família Arecaceae, dificultam o manuseio do material. Esta pesquisa teve por objetivo analisar sementes de B. maraja quanto à estrutura e qualidade fisiológica, bem como descrever a morfologia do desenvolvimento germinativo e plantular, além da morfoanatomia da plântula. Os cachos contendo frutos maduros foram coletados de 25 indivíduos, em ambiente de várzea, no município do Careiro da Várzea, Amazonas. Os pirenos e as sementes apresentaram teor de água de 23,1% e 17,9%, respectivamente. O pireno é oval a elíptico, com endocarpo rígido formado por células esclerificadas de paredes espessas, porém, ausente na região do poro germinativo. A semente é eurispérmica, sendo cordiforme, oblonga e elíptica. Apresenta dois a três poros. O tegumento é fino, formado por células esclerenquimáticas e adensadas, com conteúdo fenólico. O endosperma é homogêneo, branco e rígido. O embrião é apical, pequeno, cônico e branco-leitoso, com eixo-embrionário oblíquo e curvo, com três primórdios foliares. O endosperma armazena reserva protéica e lipídica. A temperatura alternada de 26-40 ºC foi adequada para a germinação das sementes, independente do nível de beneficiamento. A secagem progressiva do pireno facilitou a extração da semente. Contudo, a secagem e remoção do endocarpo reduziram a germinação e o vigor. Portanto, o beneficiamento não é recomendado. A embebição das sementes por dois dias favoreceu a germinação. A dormência das sementes não é causada pela presença do endocarpo. A germinação é adjacente ligulada, criptocotiledonar e hipógea. Teve início aos 152 dias após a semeadura, com a protrusão do pecíolo cotiledonar, formando o botão germinativo aos 160 dias. A plântula finalizou o desenvolvimento aos 273 com o esgotamento das reservas do haustório. O eofilo é bífido, com estômatos tetracíticos, tricomas tectores multicelulares e feixes vasculares colaterais. A raiz apresenta epiderme espessa, córtex esclerenquimático e parenquimático, endoderme conspícua; é poliarca, com cilindro vascular oco com medula esclerenquimática.

Many exogenous and endogenous compounds are referred to as endocrine disruptors (EDCs), as they interfere with natural synthesis, signaling and metabolism of endogenous hormones. Common exogenous endocrine disruptors are benzo(a)pyrene (BaP) and 17α-ethinylestradiol (EE2). Endogenous endocrine disruptor 17β-estradiol (E2) is frequently present in the environment as well. In this thesis, the effect of the mentioned EDCs and their combinations on gene and protein expression of CYP1B1, 3A1 and 3A2 in rat liver, kidney and lung was determined. Protein expression was studied using Western blot method and specific antibodies; gene expression was assessed by quantitative PCR. Moreover, the effect of tested EDCs and their combinations on BaP metabolism and CYP3A specific activity (measured as testosterone 6β-hydroxylation) were studied in liver microsomal samples. It was confirmed, that BaP significantly increases CYP1B1 expression in rat liver and lung both alone and together with EE2 or E2. Pretreatment of rat with E2 and BaP increases the ability of BaP to induce CYP1B1 expression. On the contrary, EE2, E2 and their combination decrease the CYP1B1gene expression. The rate of BaP metabolites formed in liver microsomal samples increases in rats pretreated with BaP and its combinations. In liver, there was...

Orientadores: Pedro Luiz Onófrio Volpe, Ricardo Belchior Tôrres / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Química / Made available in DSpace on 2018-08-26T22:29:35Z (GMT). No. of bitstreams: 1 Hoga_HeloisaEmi_D.pdf: 3636468 bytes, checksum: 5e4d8bd5f23dc45b0e386f59f4dd3f74 (MD5) Previous issue date: 2015 / Resumo: Propriedades excesso têm sido um caminho qualitativo e quantitativo para predizer o desvio da idealidade de soluções líquidas binárias. Além disso, as grandezas excesso têm sido usadas para desenvolver modelos e novas teorias de soluções. Neste trabalho teve como objetivo o estudo experimental de grandezas volumétricas, viscosimétricas, acústicas e das entalpias excesso de soluções líquidas binárias de água + etanol, ou + etileno glicol, ou + dietileno glicol, ou + trietileno glicol, ou + polietileno glicol 200 (PEG 200), ou + polietileno glicol 300 (PEG 300), ou + polietileno glicol 400 (PEG 400), ou + polietileno glicol 600 (PEG 600), ou + glicerol em função da composição, a diferentes temperaturas e à pressão atmosférica. Os dados de densidade e de velocidade do som foram determinados usando um densímetro e velocímetro (Anton Paar, Modelo DSA 5000). Os valores de viscosidade foram determinados usando um viscosímetro (Anton Paar, Modelo SVM 3000). Os valores da entalpia foram determinados utilizando um microcalorímetro (Thermal Activity Monitor, Titration mode, LKB2277, Thermometric). As temperaturas estudadas foram de 293,15, 298.15, 303,15 e 308,15 K. Somente a entalpia foi estudada a 298,15 K. Os resultados experimentais foram usados para calcular o volume molar excesso, o desvio da compressibilidade isentrópica, o desvio da viscosidade, a energia de Gibbs de ativação excesso e a entalpia molar excesso. As propriedades excesso foram correlacionadas usando os polinômios do tipo Redlich-Kister e Myers-Scott. Os resultados foram comparados e discutidos com base nas interações intermoleculares entre os componentes estudados / Abstract: Excess properties have been a qualitative and quantitative way to predict ideality deviation of liquid binary mixtures. Moreover the excess properties have been used to development molecular theories and new solution models. In this study, density, speed of sound, viscosity and enthalpy of binary mixtures of water + ethanol, or + ethylene glycol, or + diethylene glycol, or + triethylene glycol, or + polyethylene glycol 200, or + polyethylene glycol 300, or + polyethylene glycol 400, or + polyethylene glycol 600, or + glycerol have been determined as a function of composition at several temperatures and atmospheric pressure. The density and speed of sound data were determined using a density and sound velocity meter (Anton Paar, Model DSA 5000) The viscosity data was determined using a viscometer (Anton Paar, Model SVM 3000). The enthalpy date was determined using a microcalorimeter (Thermal Activity Monitor, Titration mode, LKB2277, Thermometric). The temperatures studied were 293.15, 298.15, 303.15 and 308.15 K. Only enthalpy data was measured at 298.15 K. The experimental results have been used to calculated the excess molar, deviations in isentropic compressibility, deviations in viscosity, excess Gibbs free energy of activation of viscous flow and excess molar enthalpy. The excess properties were correlated by using a Redlich-Kister and Myers-Scott polynomial equation. The results were discussed on the basis of intermolecular interactions between the components of the analyzed mixtures / Doutorado / Físico-Química / Doutora em Ciências

The effect of hydrophobically modified hyaluronan on surfactants aggregation has been studied in this master’s thesis. The value of critical micelle concentration of anionic surfactant SDBS (sodium dodecylbenzensulfonate), cationic surfactant CTAB (cetyltrimethylamonnium bromide) and nonionic surfactant Triton X-100 (octylphenol ethoxylate) was determined by fluorescence spectroscopy using pyrene probe. Aggregation behavior of surfactants was performed with addition of hydrophobically modified hyaluronan of two molecular weights (17 kDa, 206 kDa) in aqueous solution. The greatest influence of hydrophobized hyaluronan on aggregation behavior was observed in system with cationic surfactant CTAB. Stability of system containing cationic surfactant and hydrophobically modified hyaluronan was established through zeta potential. Last part of thesis deals with size determination using dynamic light scattering.

ABSTRACT Cytochrome b5 (CYB5) is heme protein capable of reduction of cytochromes P450 (CYP) or some other enzymes. However, his regulative capability was also observed by his apo form, i.e. in absence of heme prosthetic group in the active center. CYB5 can accept electron from cytochrome b5 reductase (CYB5R) or from cytochrome P450 reductase (CYPOR). CYPOR by itself is reduced by NADPH and is also able to forward electron to CYP independently of CYB5. CYB5R on the other hand is reduced by NADH. Efficiency of CYB5 to accept and forward an electron was studied in vitro with five different substrates - testosterone, Sudan I, aristolochic acid I (AAI), ellipticine and vandetanib. These substrates were chosen considering their characteristic reactions, which are catalyzed by their respective isoforms of CYP. The experiments with these substrates were carried out in the medium with recombinant CYPs prepared in insect cells or E. coli or in the medium with hepatic microsomes isolated from different organisms. Rats, from which the majority of these microsomes was isolated, were premedicated by different CYP inducers. The experiments were carried out in medium with NADH or NADPH in order to assess the capability of CYB5 to reduce CYP independently of CYPOR. The capability of CYB5 and CYB5R to act as a...

Dans les zones humides, un déclin dramatique des populations d’amphibiens a été observé au niveau mondial depuis les années 80. Malgré de nombreuses études suggérant que les amphibiens sont très sensibles aux pressions anthropiques cumulées, le rôle joué par les perturbateurs endocriniens dans le déclin de leurs populations reste incertain. Parmi les perturbateurs endocriniens contaminants les zones humides, le benzo[a]pyrène (BaP) et le triclosan (TCS) sont couramment retrouvés. Au cours de ce travail, nous avons étudié la réponse physiologique de xénopes (Xenopus tropicalis) exposés à court terme (24 heures) et à long terme (du têtard à l’adulte mature) au BaP et au TCS à des concentrations réalistes d’un point de vue environnemental. En combinant une approche transcriptomique à haut débit et une caractérisation des modifications cellulaires et physiologiques, nous avons mis en évidence que le BaP, le TCS et leur mélange perturbent la physiologie globale du foie. Ce trouble est caractérisé par un phénotype semblable à de l’insulino-résistance et à une stéatopathie non alcoolique (NAFLD) associé à une hépatotoxicité médiée par une altération du métabolisme lipidique. Outre les effets directs sur le métabolisme des individus exposés, il apparaît que les perturbateurs endocriniens étudiés affectent également le succès reproducteur de ces individus et le développement de leur descendance. Prises dans leur ensemble, nos observations apportent ainsi des débuts de réponse quant aux rôles de la contamination chronique des milieux sur le déclin des populations d’amphibiens. / In wetlands, dramatic decline in amphibian populations have been described worldwide since the 80s. Despite numerous studies suggesting that amphibians are highly sensitive to cumulative anthropogenic stresses, the role played by endocrine disruptors in the decline of amphibian populations remains unclear. Among the endocrine disruptors contaminating wetlands, benzo[a]pyrene (BaP) and triclosan (TCS) are recurrently found. In this work, we studied the physiological response of Xenopus tropicalis exposed to environmentally realistic concentrations of BaP and TCS during a short-term (24 hours) and long-term (from tadpole to mature adult) period. By combining a high throughput transcriptomic approach and a characterization of cellular and physiological modifications, we showed that BaP, TCS and their mixture disrupt overall liver physiology highlighted by insulin resistance-like and non-alcoholic fatty liver disease (NAFLD)-like phenotypes together with hepatotoxicity due to the impairment of lipid metabolism. Additionally to direct metabolic impact observed on exposed individuals, the studied endocrine disruptors affected their fitness and their progeny development. Altogether, our observations bring new insights into the role plays by chronic contamination of wetlands on amphibian population decline.

No description available.

Reversed-phase liquid chromatography (RPLC) is a commonly used separation technique in chemistry. Nevertheless, the mechanistic interactions at the molecular level among the eluent, analyte, and the stationary phase are not fully understood. Because of this limited understanding, optimization of the separation must be done experimentally. Learning more about molecular interactions should aid in improving separations. We are currently using second-harmonic generation (SHG) spectroscopy to investigate how analytes adsorb to the surface. SHG is a spectroscopic technique that produces signal only at places of non-isotropic symmetry; this typically occurs at surfaces. SHG can be used to produce surface isotherms of test analytes adsorbed to a model C18 stationary phase surface. Fitting these isotherms with a Langmuir model produces an adsorption equilibrium constant. However, the equilibrium constant can only be accurately determined if the true bulk concentration is known; this thesis describes an approach to ensure this. The equilibrium constant relates to Gibbs free energy and is the start to obtaining other thermodynamic information. The long equilibration times of analytes with the stationary phase observed in this study emphasize the importance of both thermodynamic information and kinetic values for understanding retention. Once equilibrium constants and other parameters are accurately obtained, this information can be used to improve predictions and calculations from numerical models.