Fast dynamics in the optical storage with Langmuir-Blodgett films of a diazocrown ether molecule

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Coloring ionic trapping states in WO3 and Nb2O5 electrochromic materials

Complex electro-optical analysis is a very useful approach to separate different kinetic processes that occur during ionic insertion reactions in electrochromic oxide materials. In this paper, we use this type of combined technique to investigate ionic and optical changes in different oxide host systems, i.e., in two oxide hosts, specifically WO3 and Nb2O5. A comparison of their electro-optical responses revealed the presence of an ionic trapping contribution to the kinetics of the coloring sites, which was named here as coloring ionic trapping state. As expected, this coloring trapping process is slower in Nb2O5 since the reduction potential of Nb2O5 is more negative (more energy is needed for a higher degree of coloration). A phenomenological solid-state model that encompasses homogeneous charge transfer and valence trapping was proposed to explain the coloring ionic trapping process. Basically the model is able to explain how ionic dynamics at low frequency region, i.e., the slower kinetic step, controls the coloring kinetics, i.e., how it is capable to regulate the coloring rates.Optical transient analyses demonstrated the possibility of the presence of more than one coloring ionic trap, indicating the complexity of the processes involved in coloration phenomenon in metal oxide host systems. (C) 2008 Published by Elsevier Ltd.

Influence from the free volume on the photoinduced birefringence in azocompound-containing polymers

The photoinduced birefringence is analyzed in a guest-host azobenzene-containing polymer in the temperature range from 20 to 330 K. An anomalous behavior arises in the low-temperature range, suggesting strong influence from the free volume for the chromophores in the polymer. This influence is so strong that quenched samples have a photoinduced signal ca. 5 times greater than the annealed ones at room temperature. An extended free volume model is presented based on two assumptions about thermal fluctuations in the cavities and their size distribution. This model, which is an extension of the model by Mita et al., can explain the main features of the photoinduced birefringence as a function of time, temperature, and initial free volume state. To account for the influence of free volume on the photoorientation, the detailed reorientation model by Sekkat's was used. We show that Sekkat's model leads to an exponential behavior at small orientation regimes, which simplifies the mathematical treatment and allows the mean free volume to be obtained from the data fitting.

Active and exo-site inhibition of human factor Xa: Structure of des-Gla factor Xa inhibited by NAP5, a potent nematode anticoagulant protein from Ancylostoma caninum

Hookworms are hematophagous nematodes capable of growth, development and subsistence in living host systems such as humans and other mammals. Approximately one billion, or one in six, people worldwide are infected by hookworms causing gastrointestinal blood loss and iron deficiency anemia. The hematophagous hookworm Ancylostoma caninum produces a family of small, disulfide-linked protein anticoagulants (75-84 amino acid residues). One of these nematode anticoagulant proteins, NAP5, inhibits the amidolytic activity of factor Xa (fXa) with K-i = 43 pM, and is the most potent natural fXa inhibitor identified thus far. The crystal structure of NAP5 bound at the active site of gamma-carboxyglutamic acid domainless factor Xa (des-fXa) has been determined at 3.1 angstrom resolution, which indicates that Asp189 (fXa, S1 subsite) binds to Arg40 (NAP5, P1 site) in a mode similar to that of the BPTI/trypsin interaction. However, the hydroxyl group of Ser39 of NAP5 additionally forms a hydrogen bond (2.5 angstrom) with His57 NE2 of the catalytic triad, replacing the hydrogen bond of Ser195 OG to the latter in the native structure, resulting in an interaction that has not been observed before. Furthermore, the C-terminal extension of NAP5 surprisingly interacts with the fXa exosite of a symmetry-equivalent molecule forming a short intermolecular beta-strand as observed in the structure of the NAPc2/fXa complex. This indicates that NAP5 can bind to fXa at the active site, or the exosite, and to fX at the exosite. However, unlike NAPc2, NAP5 does not inhibit fVIIa of the fVIIa/TF complex. (c) 2007 Elsevier Ltd. All rights reserved.

Alternatives for the Treatment of Schistosomiasis: Physico-Chemical Characterization of an Inclusion Complex Between Praziquantel and Hydroxypropyl-beta-Cyclodextrin

"Alternatives for the Treatment of Schistosomiasis: Physico-Chemical Characterization of an Inclusion Complex Between Praziquantel and Hydroxypropyl-beta-Cyclodextrin". Praziquantel (PZQ) is the drug of choice commonly used for the treatment of shistosomiasis. However, it has low aqueous solubility, which could limit its bioavailability in the body. To circumvent these features, an inclusion complex with hydroxypropyl-beta-cyclodextrin (HP-beta-CD) was prepared. Thus, the objective of this work was to prepare and characterize the PZQ/HP-beta-CD inclusion complex. Morphological, spectroscopic, and calorimetric analysis showed the first signs of the guest/host interaction. The complexation kinetic analysis was used to determine the kinetic constant and, besides that, it was possible to establish the time consumed to reach equilibrium. Using the solubility isotherm, it was observed that the interaction with HP-beta-CD increased 2.4 fold the aqueous solubility of plain PZQ. In vitro cytotoxicity tests, using fibroblast cells, evidenced no toxicity for these cells at the concentrations tested. These results demonstrated that there is a potential use of PZQ in formulations with HP-beta-CD.

Alternativas para o tratamento da esquistossomose: Caracterizacao fisico-quimica do complexo de inclusao entre praziquantel e hidroxipropil-β-ciclodextrina

Praziquantel (PZQ) is the drug of choice commonly used for the treatment of shistosomiasis. However, it has low aqueous solubility, which could limit its bioavailability in the body. To circumvent these features, an inclusion complex with hydroxypropyl-beta- cyclodextrin (HP-β-CD) was prepared. Thus, the objective of this work was to prepare and characterize the PZQ/HP-β-CD inclusion complex. Morphological, spectroscopic, and calorimetric analysis showed the first signs of the guest/host interaction. The complexation kinetic analysis was used to determine the kinetic constant and, besides that, it was possible to establish the time consumed to reach equilibrium. Using the solubility isotherm, it was observed that the interaction with HP-β-CD increased 2.4 fold the aqueous solubility of plain PZQ. In vitro cytotoxicity tests, using fibroblast cells, evidenced no toxicity for these cells at the concentrations tested. These results demonstrated that there is a potential use of PZQ in formulations with HP-β-CD.

Clonagem e expressão do gene da nucleoproteína do vírus da bronquite infecciosa em sistemas hospedeiros eucarioto (Pichia pastoris) e procarioto (Escherichia coli)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Estudo da orientação dipolar fotoassistida de grupos azobenzênicos em filmes finos através de medidas da atividade eletro-óptica

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Increasing Doxycycline Hyclate Photostability by Complexation with beta-Cyclodextrin

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Photoinduced orientation in natural rubber

Azobenzene molecules and their derivatives have been widely investigated for their potential applications in optical and electrooptical devices. We have prepared a new guest-host system from natural rubber (NR) impregnated with azobenzene derivative Sudan Red B (SRB). The effects of stretching and immersion time on photoinduced orientation were investigated by birefringence signal measurements. We have found that the molecular orientation increase when the samples are stretched and decrease with the increase of immersion time. The first behavior was explained by using the random coil model and the latter was attributed to increase of the aggregation of SRB into NR matrix. (C) 2012 Published by Elsevier B.V.

Untersuchung molekularer Erkennungsreaktionen mit einem integriert-optischen Mach-Zehnder-Interferometer

Abstract (deutsch)Zielsetzung des Dissertationsvorhabens war die Beobachtung und Analyse von Gast-Wirt-Wechselwirkungen an oxidischen Oberflächen. Einer der Wechselwirkungspartner sollte dabei auf der Oberfläche immobilisiert, der andere in wäßriger Lösung darüber vorliegen.Eine empfindliche und oberflächensensitive Methode zur Beobachtung der Anlagerung unmarkierter Moleküle ist die Wellenleiterspektroskopie, insbesondere mit dem hier verwendeten und weiterentwickelten integriert-optischen Mach-Zehnder-Interferometer in Siliziumtechnik (Siliziumoxynitrid auf oxidiertem Siliziumwafer). Mit Hilfe des Interferometers wurden unterschiedliche Wirt-Gast-Systeme untersucht. Grundlage der Immobilisierung war jeweils die Funktionalisierung der Sensoroberfläche durch Selbstadsorption von Organosilanen. Durch unterschiedliche Organosilane, die zum Teil im Rahmen dieser Arbeit synthetisiert wurden, ließen sich die Wirtmoleküle beta-Cyclodextrin, Streptavidin, sowie unterschiedliche monoklonale Antikörperfragmente immobilisieren.- Der Einfluß der Oberfläche auf die Bindungsstärke des Wirtmoleküls beta-Cyclodextrin und unterschiedlicher Gastmoleküle wurde konzentrationsabhängig untersucht.- Silan-Biotinderivate mit unterschiedlicher Streptavidin-Affinität wurden an die Oberfläche immobilisiert und die Adsorption von Streptavidin an die Biotinderivate beobachtet. Dabei konnte unter anderem nachgewiesen werden, daß das Streptavidinadsorbat gequollen ist.- Als mögliche Anwendung wurde geprüft, ob das vorgestellte Interferometer durch die Funktionalisierung mit Antikörperfragmenten als Biosensor in Frage kommt. Es konnte nachgewiesen werden, daß sich Antikörper auf der Sensoroberfläche immobilisieren lassen und Antigene spezifisch an diese Antikörper adsorbieren.

The Synthesis and properties of functionalised polymers

The aim of this project was to synthesise fluorinated polymers that might act as hot material in a guest-host system for use in non-linear optical applications. These polymers would be expected to have the advantage over materials such as poly(vinylidene fluoride) which is known to be incompatible with many nlo active materials. A series of bicyclic fluorinated monomers was prepared by the reaction of fluorinated dienophiles with cyclopentadiene in a series of Diels-Alder reactions. The monomers were purified and then used in ring opening metathesis polymerisation. The materials were then characterised by gel permeation chromatography and nuclear magnetic resonance spectroscopy and cast as films for determination of their activities as nlo materials using a Nd/YAG laser system. The second harmonic intensity of each was measured relative to quartz. However no materials of significant activity were produced. In an attempt to produce polymers that might subsequently be functionalised the polymerisation of 1,2-methylenedioxybenzene and 1,4-benzodioxane was investigated.

Playing catch-up with Escherichia coli:using yeast to increase success rates in recombinant protein production experiments

Several host systems are available for the production of recombinant proteins, ranging from Escherichia coli to mammalian cell-lines. This article highlights the benefits of using yeast, especially for more challenging targets such as membrane proteins. On account of the wide range of molecular, genetic, and microbiological tools available, use of the well-studied model organism, Saccharomyces cerevisiae, provides many opportunities to optimize the functional yields of a target protein. Despite this wealth of resources, it is surprisingly under-used. In contrast, Pichia pastoris, a relative new-comer as a host organism, is already becoming a popular choice, particularly because of the ease with which high biomass (and hence recombinant protein) yields can be achieved. In the last few years, advances have been made in understanding how a yeast cell responds to the stress of producing a recombinant protein and how this information can be used to identify improved host strains in order to increase functional yields. Given these advantages, and their industrial importance in the production of biopharmaceuticals, I argue that S. cerevisiae and P. pastoris should be considered at an early stage in any serious strategy to produce proteins.

Membrane protein production in the Yeast, S. cerevisiae

The first crystal structures of recombinant mammalian membrane proteins were solved in 2005 using protein that had been produced in yeast cells. One of these, the rabbit Ca2+-ATPase SERCA1a, was synthesized in Saccharomyces cerevisiae. All host systems have their specific advantages and disadvantages, but yeast has remained a consistently popular choice in the eukaryotic membrane protein field because it is quick, easy and cheap to culture, whilst being able to post-translationally process eukaryotic membrane proteins. Very recent structures of recombinant membrane proteins produced in S. cerevisiae include those of the Arabidopsis thaliana NRT1.1 nitrate transporter and the fungal plant pathogen lipid scramblase, TMEM16. This chapter provides an overview of the methodological approaches underpinning these successes.