Design of a Modern Liposome and Bee Venom Formulation for the Traditional VIT-Venom Immunotherapy

Traditional venom immunotherapy uses injections of whole bee venom in buffer or adsorbed in Al (OH)(3) in an expensive, time-consuming way. New strategies to improve the safety and efficacy of this treatment with a reduction of injections would, therefore, be of general interest. It would improve patient compliance and provide socio-economic benefits. Liposomes have a long tradition in drug delivery because they increase the therapeutic index and avoid drug degradation and secondary effects. However, bee venom melittin (Mel) and phospholipase (PLA(2)) destroy the phospholipid membranes. Our central idea was to inhibit the PLA(2) and Mel activities through histidine alkylation and or tryptophan oxidation (with pbb, para-bromo-phenacyl bromide, and/or NBSN-bromosuccinimide, respectively) to make their encapsulations possible within stabilized liposomes. We strongly believe that this formulation will be nontoxic but immunogenic. In this paper, we present the whole bee venom conformation characterization during and after chemical modification and after interaction with liposome by ultraviolet, circular dichroism, and fluorescence spectroscopies. The PLA(2) and Mel activities were, measured indirectly by changes in turbidity at 400(nm), rhodamine leak-out, and hemolysis. The native whole bee venom (BV) presented 78.06% of alpha-helical content. The alkylation (A-BV) and succynilation (S-BV) of BV increased 0.44 and 0.20% of its alpha-helical content. The double-modified venom (S-A-BV) had a 0.74% increase of alpha-helical content. The BV chemical modification induced another change on protein conformations observed by Trp that became buried with respect to the native whole BV. It was demonstrated that the liposomal membranes must contain pbb (SPC:Cho:pbb, 26:7:1) as a component to protect them from aggregation and/or fusion. The membranes containing pbb maintained the same turbidity (100%) after incubation with modified venom, in contrast with pbb-free membranes that showed a 15% size decrease. This size decrease was interpreted as membrane degradation and was corroborated by a 50% rhodamine leak-out. Another fact that confirmed our interpretation was the observed 100% inhibition of the hemolytic activity after venom modification with pbb and NBS (S-A-BV). When S-A-BV interacted with liposomes, other protein conformational changes were observed and characterized by the increase of 1.93% on S-A-BV alpha-helical content and the presence of tryptophan residues in a more hydrophobic environment. In other words, the S-A-BV interacted with liposomal membranes, but this interaction was not effective to cause aggregation, leak-out, or fusion. A stable formulation composed by S-A-BV encapsulated within liposomes composed by SPC:Cho:pbb, at a ratio of 26:7:1, was devised. Large unilamellar vesicles of 202.5 nm with a negative surface charge (-24.29 mV) encapsulated 95% of S-A-BV. This formulation can, now, be assayed on VIT.

Preparation and characterization of the novels terpolymers of poly-{trans-[RuCl(2)(vpy)(4)]-styrene-divinylbenzene} and styrene-divinylbenzene-vinylpiridine impregnated with silver nanoparticles

This paper reports the preparation and characterization of poly-{trans-[RuCl(2)(vpy)(4)]-styrene-divinylbenzene} and styrene-divinylbenzene-vinylpiridine filled with nanosilver. Theses materials were synthesized by non aqueous polymerization through a chemical reaction using benzoyl peroxide as the initiator. The nanosilver was obtained from chemical reduction using NaBH(4) as reducing agent and sodium citrate as stabilizer. The nanometric dimension of nanosilver was monitored by UV-visible and confirmed through TEM. The morphology was characterized by SEM and the thermal properties were done by TGA and DSC. The antimicrobial action of the polymers impregnated with nanosilver was evaluated using both microorganisms, Staphylococcus aureus and Escherichia coli. The antimicrobial activity of the poly-{trans-[RuCl(2)(vpy)(4)]-styrene-divinylbenzene} filled with nanosilver was confirmed by the presence of an inhibition halo of the bacterial growth in seeded culture media, but was not confirmed to the styrene-divinylbenzene-vinylpiridine. The present work suggest that trans - [RuCl(2)(vpy)(4)] complex facilitate the release of silver ion from the media.

Spectroscopic characterization of the structural changes of polyaniline nanofibers after heating

The thermal behavior of PANI nanofibers doped with beta-naphthalenesulfonic acid (beta-NSA) was investigated and their morphological and structural changes after heating were monitored by SEM, XRD and Raman techniques, respectively. By using electron-scanning microscopy it is possible to verify that the nanofiber morphology is stable and no polymer degradation is observed in thermogravimetric (TG) data up to 200 degrees C. Nevertheless, the heating promotes the formation of cross-linking structures (phenazine and/or oxazine-like rings), that is clearly demonstrated by the presence of bands at ca. 578, 1398, and 1644 cm(-1) in resonance Raman spectra of heated PANI-NSA samples. The most important consequence of the formation of cross-linking structures in PANI-NSA samples is that these samples retain their nanofiber morphology upon HCl doping in contrast to PANI-NSA nanofibers without heating. (c) 2007 Elsevier Ltd. All rights reserved.

Bacterial cellulose-laponite clay nanocomposites

A novel material comprised of bacterial cellulose (BC) and Laponite clay with different inorganic organic ratios (m/m) was prepared by the contact of never-dried membranes of BC with a previous dispersion of clay particles in water. Field emission scanning electron microscopy (FE-SEM) data of composite materials revealed an effective adhesion of clay over the surface of BC membrane; inorganic particles also penetrate into the polymer bulk, with a significant change of the surface topography even at 5% of clay loading. As a consequence, the mechanical properties are deeply affected by the presence of clay, increasing the values of the Young modulus and the tensile strength. However the maximum strain is decreased when the clay content is increased in the composite in comparison to pristine BC. The main weight loss step of the composites is shifted towards higher temperatures compared to BC, indicating that the clay particles slightly protect the polymer from thermal and oxidative decomposition. (C) 2010 Elsevier Ltd. All rights reserved.

On the apparent lack of preferential site occupancy and electrooxidation of CO adsorbed at low coverage onto stepped platinum surfaces

We report time evolution studies of low coverage CO adsorption (surface hydrogen site blocking < 40%) and oxidative stripping on stepped Pt(776) and Pt(554) surfaces. It was observed that there is no preferential site occupancy for CO adsorption on step or terrace. It is proposed that CO adsorption onto these surfaces is a random process, and after CO adsorption there is no appreciable shift from CO-(111) to CO-(110) sites. This implies that after adsorption, CO molecules either have a very long residence time, or that the diffusion coefficient is much lower than previously thought. After CO electrooxidation the sites released included both terrace (111) and step (110) orientations. For surface hydrogen site blocking > 40%, the lateral interactions might play a role in the preferential CO site occupancy. (C) 2011 Elsevier B.V. All rights reserved.

Thermal energy storage in Swedish single family houses : a case study

In a Nordic climate, space heating (SH) and domestic hot water (DHW) used in buildings constitute a considerable part of the total energy use in the country. For 2010, energy used for SH and DHW amounted to almost 90 TWh in Sweden which corresponds to 60 % of the energy used in the residential and service sector, or almost 24 % of the total final energy use for the country. Storing heat and cold with the use of thermal energy storage (TES) can be one way of increasing the energy efficiency of a building by opening up possibilities for alternative sources of heat or cold through a reduced mismatch between supply and demand. Thermal energy storage without the use of specific control systems are said to be passive and different applications using passive TES have been shown to increase energy efficiency and/or reduce power peaks of systems supplying the heating and cooling needs of buildings, as well as having an effect on the indoor climate. Results are however not consistent between studies and focus tend to be on the reduction of cooling energy or cooling power peaks. In this paper, passive TES introduced through an increased thermal mass in the building envelope to two single family houses with different insulation standard is investigated with building energy simulations. A Nordic climate is used and the focus of this study is both on the reduction of space heating demand and space heating power, as well as on reduction of excess temperatures in residential single family houses without active cooling systems. Care is taken to keep the building envelope characteristics other than the thermal mass equal for all cases so that any observations made can be derived to the change in thermal mass. Results show that increasing the sensible thermal mass in a single family house can reduce the heating demand only slightly (1-4 %) and reduce excess temperatures (temperatures above 24 degrees C) by up to 20 %. Adding a layer of PCM (phase change materials) to the light building construction can give similar reduction in heating demand and excess temperatures, however the phase change temperature is important for the results.

Filmes finos dielétricos para dispositivos microeletrônicos avançados

Apresentamos mecanismos de formação e de degradação térmica de filmes fi- nos (espessura da ordem de 10 nm) de diferentes dielétricos sobre substrato de silício monocristalino. Tendo em vista a aplicação dessas estruturas em MOSFETs (transistores de efeito de campo metal-óxido-semicondutor), estudamos o consagrado óxido de silício (SiO2), os atuais substitutos oxinitretos de silício (SiOxNy) e o possível substituto futuro óxido de alumínio (Al2O3). Nossos resultados experimentais baseiam-se em técnicas preparativas de substituição isotópica e de caracterização física com feixes de íons (análise com reações nucleares) ou raios- X (espectroscopia de fotoelétrons). Observamos que: (a) átomos de silício não apresentam difusão de longo alcance (além de ~ 2 nm) durante o crescimento de SiO2 por oxidação térmica do silício em O2; (b) nitretação hipertérmica é capaz de produzir filmes finos de oxinitreto de silício com até dez vezes mais nitrogênio que o resultante do processamento térmico usual, sendo que esse nitrogênio tende a se acumular na interface SiOxNy/Si; e (c) átomos de oxigênio, alumínio e silício migram e promovem reações químicas durante o recozimento térmico de estruturas Al2O3/SiO2/Si em presença de O2. Desenvolvemos um modelo de difusão-reação que poderá vir a permitir o estabelecimento de condições ótimas de processamento térmico para filmes finos de Al2O3 sobre silício a serem empregados na fabricação de MOSFETs.

Effect of the estufagem process on the chemical constituents of Madeira wines

Madeira wine is a product of well-established reputation, whose aroma and flavour is the result of unique combinations. Particularly, its maturation may include estufagem, wherein wine is usually heated at 45 °C for three months. During this period, several chemical changes may occur, so it is essential to assess its impact on the wine. In this sense, the main objective of the thesis was to evaluate the effect estufagem on the chemical constituents of Madeira wine, specifically on those molecules potentially important in the development of its typical features. Firstly, analytical methodologies capable of determining the target compounds, combining precision and reproducibility to execution effectiveness, were developed. Then various monovarietal Madeira wines were analysed during estufagem under standard and overheating conditions in order to assess its effect. The following compounds were evaluated: furans, amino acids, biogenic amines, polyphenols, organic acids and volatile compounds. In addition, the total polyphenolic composition, the antioxidant potential and the colour of these wines were also evaluated. The results show that most constituents change due to the heating process. Particularly, the heating promotes the development of 5-hydroxymethylfurfural (HMF) in sweet wines submitted to estufagem at higher temperatures. Moreover, estufagem provides the decrease of most amino acids, suggesting their involvement in the formation of the bouquet of these wines. Regarding the total polyphenol content and antioxidant potential of these wines they do not seem to be greatly affected by the heating step, however most monomeric polyphenols decrease during this process. The thermal processing of Madeira wines favours the development of the volatile composition, especially of volatiles usually reported as typical aromas of Madeira wines. Finally, it was demonstrated that the thermal degradation of sugars, especially of fructose, promotes the emergence of volatile compounds identified in baked wines.

Caracterização Microestrutural e Tribocorrosiva de Sistemas Metalocerâmicos Odontológicos do Tipo Ni-Cr/Porcelana E Ni-Cr/Ti/Porcelana

The partial fixed prosthodontics restoration is used to rehabilitate form and function of partial or total compromised teeth, having to remain permanently joined to remainder tooth. The most useful material on prosthodontics is the feldspar porcelain, commercialized as aluminosilicate powders. Dental porcelains are presented with limited mechanical properties to rehabilitate extensive spaces. The association with Ni-Cr metallic systems (metal-ceramic system) allows that the metallic substructure compensates the fragile porcelain nature, preserving the thermal insulation and aesthetics desirable, as well as reducing the possibility of cracking during matication efforts. Cohesive flaws by low mechanical strength connect the metallic substructure to the oral environment, characterized by a electrolytic solution (saliva), by aggressive temperature, pH cyclic changes and mechanical requests. This process results on ionic liberation that could promote allergic or inflammatory responses, and/or clinical degradation of ceramometal system. The aim of this study was to evaluate the presence of an intermediate titanium layer on the microscopic fracture behavior of porcelains on ceramometal systems. Plasma deposition of titanium films result in regular passivating oxide layers which act as barriers to protect the metallic substrate against the hazardous effects of corrosive saliva. Tribocorrosion tests were performed to simulate the oral environment and mechanical stress, making it possible the early detection of crack formation and growth on metal-ceramic systems, which estimate the adherence between the compounds of this system. Plain samples consisting of dental feldspar porcelain deposited either onto metallic substrates or titanium films were fired and characterized by scanning electron microscopy. The result showed that the titanium film improved the adherence of the system compared to conventional metal-ceramic interfaces, thus holding crack propagation

Estudo da sinterização de eletrólito sólido de céria dopada com gadolínia

Fuel cells are electrochemical devices that convert chemical energy in electrical energy by a reaction directly. The solid oxide fuel cell (SOFC) works in temperature between 900ºC up to 1000ºC, Nowadays the most material for ceramic electrolytes is yttria stabilized zirconium. However, the high operation temperature can produce problems as instability and incompatibility of materials, thermal degradation and high cost of the surround materials. These problems can be reduced with the development of intermediate temperature solid oxide fuel cell (IT-SOFC) that works at temperature range of 600ºC to 800ºC. Ceria doped gadolinium is one of the most promising materials for electrolytes IT-SOFC due high ionic conductivity and good compatibility with electrodes. The inhibition of grain growth has been investigated during the sintering to improve properties of electrolytes. Two-step sintering (TSS) is an interesting technical to inhibit this grain growth and consist at submit the sample at two stages of temperature. The first one stage aims to achieve the critical density in the initiating the sintering process, then the sample is submitted at the second stage where the temperature sufficient to continue the sintering without accelerate grain growth until to reach total densification. The goal of this work is to produce electrolytes of ceria doped gadolinium by two-step sintering. In this context were produced samples from micrometric and nanometric powders by two routes of two-step sintering. The samples were obtained with elevate relative density, higher than 90% using low energy that some works at the same area. The average grain size are at the range 0,37 μm up to 0,51 μm. The overall ionic conductivity is 1,8x10-2 S.cm and the activation energy is 0,76 eV. Results shown that is possible to obtain ceria-doped gadolinium samples by two-step sintering technique using modified routes with characteristics and properties necessary to apply as electrolytes of solid oxide fuel cell

Adição de poliuretana em pastas de cimento para poços de petróleo como agente de correção do filtrado

Chemical admixtures, when properly selected and quantified, play an important role in obtaining adequate slurry systems for quality primary cementing operations. They assure the proper operation of a well and reduce costs attributed to corrective cementing jobs. Controlling the amount lost by filtering through the slurry to permeable areas is one of the most important requirements in an operation, commonly controlled by chemical admixtures, such as carboxymethylcellulose (CMC). However, problems related to temperature, salttolerance and the secundary retarding effect are commonly reported in the literature. According to the scenario described above, the use of an aqueous dispersion of non-ionic poliurethane was proposed to control the filter loss, given its low ionic interaction with the free ions present in the slurries in humid state. Therefore, this study aims at assessing the efficiency of poliurethane to reduce filter loss in different temperature and pressure conditions as well as the synergistic effect with other admixtures. The temperatures and pressures used in laboratory tests simulate the same conditions of oil wells with depths of 500 to 1200 m. The poliurethane showed resistance to thermal degradation and stability in the presence of salts. With the increase in the concentration of the polymer there was a considerable decrease in the volume lost by filtration, and this has been effective even with the increase in temperature

Degradacao termica e catalitica da borra oleosa de Petroleo com materiais nanoestruturados al-mcm-41 e AL-SBA-15

Aiming to reduce and reuse waste oil from oily sludge generated in large volumes by the oil industry, types of nanostructured materials Al-MCM-41 and Al-SBA-15, with ratios of Si / Al = 50, were synthesized , and calcined solids used as catalysts in the degradation of oily sludge thermocatalytic oil from oilfield Canto do Amaro, in the state of Rio Grande do Norte. Samples of nanostructured materials were characterized by thermogravimetric analysis (TG / DTG), X-ray diffraction (XRD), scanning electron microscopy (SEM), absorption spectroscopy in the infrared Fourier transform (FT-IR) and adsorption nitrogen (BET). The characterization showed that the synthesized materials resulted in a catalyst nanostructure, and ordered pore diameter and surface area according to existing literature. The oily sludge sample was characterized by determining the API gravity and sulfur content and SARA analysis (saturates, aromatics, resins and asphaltenes). The results showed a material equivalent to the average oil with API gravity of 26.1, a low sulfur content and considerable amount of resins and asphaltenes, presented above in the literature. The thermal and catalytic degradation of the oily sludge oil was performed from room temperature to 870 ° C in the ratios of heating of 5, 10 and 20 ° C min-1. The curves generated by TG / DTG showed a more accelerated degradation of oily sludge when it introduced the nanostructured materials. These results were confirmed by activation energy calculated by the method of Flynn-Wall, in the presence of catalysts reduced energy, in particular in the range of cracking, showing the process efficiency, mainly for extraction of lightweight materials of composition of oily sludge, such as diesel and gasoline

COMBUSTION and PYROLYSIS of A SLUDGE FROM WASTEWATER TREATMENT PLANT

The possibility of thermal treatment plants of municipal wastewater is an alternative solution for the final disposition of the sludge produced on small cities as Barueri, a small town of São Paulo State, Brazil. Combustion and pyrolysis of that municipal waste, occurring respectively in air and nitrogen, have been studied by thermogravimetry (TG) and differential thermal analysis (DTA). The main steps of each case were analyzed and Kissinger plots were used to estimate respective activation energies. DTG peaks are more indicated to represent the condition of maximum reaction rates than DTA peaks.

Nanotechnological delivery systems for the oral administration of active molecules: Polymeric microparticles and microemulsions applied to anti-inflammatory and anti-infectious drugs

This thesis was devoted to the development of innovative oral delivery systems for two different molecules. In the first part, microparticles (MPs) based on xylan and Eudragit® S- 100 were produced and used to encapsulate 5-aminosalicylic acid for colon delivery. Xylan was extracted from corn cobs and characterized in terms of its physicochemical, rheological and toxicological properties. The polymeric MPs were prepared by interfacial cross-linking polymerization and spray-drying and characterized for their morphology, mean size and distribution, thermal stability, crystallinity, entrapment efficiency and in vitro drug release. MPs with suitable physical characteristics and satisfactory yields were prepared by both methods, although the spray-dried systems showed higher thermal stability. In general, spraydried MPs would be preferable systems due to their thermal stability and absence of toxic agents used in their preparation. However, drug loading and release need to be optimized. In the second part of this thesis, oil-in-water microemulsions (O/W MEs) based on mediumchain triglycerides were formulated as drug carriers and solubility enhancers for amphotericin B (AmB). Phase diagrams were constructed using surfactant blends with hydrophiliclipophilic balance values between 9.7 and 14.4. The drug-free and drug-loaded MEs presented spherical non-aggregated droplets around 80 and 120 nm, respectively, and a low polydispersity index. The incorporation of AmB was high and depended on the volume fraction of the disperse phase. These MEs did not reduce the viability of J774.A1 macrophage-like cells for concentrations up to 25 μg/mL of AmB. Therefore, O/W MEs based on propylene glycol esters of caprylic acid may be considered as suitable delivery systems for AmB

Estudo do efeito dos metabólitos do benzeno e tolueno sobre as mitocôndris cerebrais e hepáticas de ratos

Aim: The aim of this work was to investigate the hypothesis that catechol and 3MC inhibit FADH2-linked basal respiration in mitochondria isolated from rat liver and brain homogenates. Moreover, catechol ability to induce DNA damage in rat brain cells through the comet assay (alkaline single-cell gel electrophoresis assay) was also observed. Methods: Two different catechols were evaluated: pirocatechol (derived from benzene) and 3-methylcatechol (derived from toluene); rat liver and brain homogenates were incubated with 1mM catechol at pH 7.4 for up to 30 minutes. After that, mitochondrial fractions were isolated by differential centrifugation. Basal oxygen uptake was measured using a Clark-type electrode after the addition of 10 mM sodium succinate for a period of 12 minutes. In additional experiments, rat brain cells were treated with 1, 5 and 10mM pirocatechol for up to 20 minutes at 37º C, and submitted to electrophoresis. Results: Catechols (pirocatechol and 3methylcatechol) induced a time-dependent partial inhibition of FADH2-linked basal mitochondrial respiration. Indeed, pirocatechol was able to produce a dosedependent DNA oxidative damage in rat brain cells by 2 and 4 injury levels. These results suggest that reactive oxygen species generated by the oxidation of catechols, induced an impairment on mitochondrial respiration and a DNA damage, which might be related to their citotoxicity. Conclusion: Catechols produced an inhibition of basal respiration associated to FADH2 in isolated liver and brain mitochondria; 3-methylcatechol, at the same concentration, produced similar toxicity in the mitochondrial model. Indeed, pirocatechol induced a DNA damage in rat brain cells, mainly observed in comets formation and consequent DNA degradation