Oxindole-Schiff base copper(II) complexes interactions with human serum albumin: Spectroscopic, oxidative damage, and computational studies

CD and EPR were used to characterize interactions of oxindole-Schiff base copper(II) complexes with human serum albumin (HSA). These imine ligands form very stable complexes with copper, and can efficiently compete for this metal ion towards the specific N-terminal binding site of the protein, consisting of the amino acid sequence Asp-Ala-His. Relative stability constants for the corresponding complexes were estimated from CD data, using the protein as competitive ligand, with values of log K(CuL) in the range 15.7-18.1, very close to that of [Cu(HSA)] itself, with log K(CuHSA) 16.2. Some of the complexes are also able to interfere in the a-helix structure of the protein, while others seem not to affect it. EPR spectra corroborate those results, indicating at least two different metal species in solution, depending on the imine ligand. Oxidative damage to the protein after incubation with these copper(II) complexes, particularly in the presence of hydrogen peroxide, was monitored by carbonyl groups formation, and was observed to be more severe when conformational features of the protein were modified. Complementary EPR spin-trapping data indicated significant formation of hydroxyl and carbon centered radicals, consistent with an oxidative mechanism. Theoretical calculations at density functional theory (DFT) level were employed to evaluate Cu(II)-L binding energies, L -> Cu(II) donation, and Cu(II) -> L back-donation, by considering the Schiff bases and the N-terminal site of HSA as ligands. These results complement previous studies on cytotoxicity, nuclease and pro-apoptotic properties of this kind of copper(II) complexes, providing additional information about their possibilities of transport and disposition in blood plasma. (C) 2009 Elsevier Inc. All rights reserved.

The role of oxygen in the interaction of emeraldine base polyaniline with Cu(II) or Fe(III) ions in NMP solution

The influence of molecular oxygen in the interactions of emeraldine base form of polyaniline (EB-PANI) with Fe(III) or Cu(II) ions in 1-methyl-2-pyrrolidinone (NMP) solutions has been investigated by UV-vis-NIR, resonance Raman and electron paramagnetic resonance (EPR) spectroscopies. Through the set of spectroscopic results it was possible to rationalize the role Of O(2) and to construct a scheme of preferential routes occurring in the interaction of EB-PANI with Fe(III) or Cu(II). Solutions of 4.0 mmol L(-1) EB-PANI with 0.8, 2.0 and 20 mmol L(-1) Fe(III) or Cu(II) ions in NMP were investigated and the main observed reactions were EB-PANI oxidation to pernigraniline (PB-PANI) and EB-PANI doping process by pseudo-protonation, or by a two-step redox process. In the presence Of O(2), PB-PANI is observed in all Fe(III)/EB solutions and EB-PANI doping only occurs in solutions with high Fe(III) concentrations through pseudo-protonation. On the other hand, emeraldine salt (ES-PANI) is formed in all Fe(III)/EB solutions under N(2) atmosphere and, in this case, doping occurs both by the pseudo-protonation and two-step redox mechanisms. In all Cu(II)/EB solutions PB-PANI is formed both in the presence and absence of O(2), and only for solutions with high Cu(II) concentrations doping process occurs in a very low degree. The most important result from EPR spectra was providing evidence for redox steps. The determined Cu(II) signal areas under oxygen are higher than under N(2) and, further. the initial metal proportions (1:2:20) are maintained in these spectra, indicating that Cu(I) formed are re-oxidized by O(2) and. so, Cu(II) ions are being recycled. Consistently, for the solutions prepared under nitrogen, the corresponding areas and proportions in the spectra are much lower, confirming that a partial reduction of Cu(II) ions actually occurs. (C) 2009 Elsevier B.V. All rights reserved.

Activation Energies of the Electrooxidation of Formic Acid on Pt(100)

The electrooxidation of small organic molecules on platinum surfaces usually involves different structure-dependent steps that include adsorption and desorption of various species and multiple reaction pathways. Because temperature plays a decisive role on each individual step, understanding its global influence on the reaction mechanism is often a difficult task, especially when the system is studied under far from equilibrium conditions in the presence of kinetic instabilities. Aiming at contributing to unravel this problem, herein, we report an experimental study of the role played by temperature on the electrooxidation of formic acid on a Pt(100) electrode. The system was investigated under both close and far from equilibrium conditions, and apparent activation energies were estimated using different strategies. Overall, comparable activation energies were estimated under oscillatory and quasi-stationary conditions, at high potentials. At low potentials, the poisoning process associated with the formic acid dehydration step presented a negligible dependence with temperature and, therefore, zero activation energy. On the basis of our experimental findings, we suggest that formic acid dehydration is the main, but maybe not the unique, step that differentiates the temperature dependence of the oscillatory electrooxidation of formic acid on Pt(100) with that on polycrystalline platinum.

Bonding agent underneath sealant: shear bond strength to oil-contaminated

This study evaluated in vitro the shear bond strength of a resin-based pit-and-fissure sealant (Fluroshield - F) associated with either an ethanol-based (Adper Single Bond 2 - SB) or an acetone-based (Prime & Bond - PB) adhesive system under conditions of oil contamination. Mesial and distal enamel surfaces from 30 sound third molars were randomly assigned to 2 groups (n=30): I - no oil contamination; II - oil contamination. Contamination (0.25 mL during 10 s) was performed after 37% phosphoric acid etching with an air/oil spray. The specimens were randomly assigned to subgroups, according to the bonding protocol adopted: subgroup A - F was applied to enamel without an intermediate bonding agent layer; In subgroups B and C, SB and PB, respectively, were applied, light-cured, and then F was applied and light-cured. Shear bond strength was tested at a crosshead speed of 0.5 mm/min in a universal testing machine. Means (± SD) in MPa were: IA-11.28 (±1.84); IIA-12.02 (±1.15); IB-9.73 (±2.38); IIB-9.62 (±2.29); IC-28.30 (±1.63); and IIC-25.50 (±1.91). It may be concluded that the oil contamination affected negatively the sealant bonding to enamel and the acetone-based adhesive system (PB) layer applied underneath the sealant was able to prevent its deleterious effects to adhesion.

Influence of incorporation of fluoroalkyl methacrylates on roughness and flexural strength of a denture base acrylic resin

Fluorinated denture base acrylic resins can present more stable physical properties when compared with conventional polymers. This study evaluated the incorporation of a fluoroalkyl methacrylate (FMA) mixture in a denture base material and its effect on roughness and flexural strength. A swelling behavior assessment of acrylic resin specimens (n=3, per substance) after 12 h of FMA or methyl methacrylate (MMA) immersion was conducted to determine the solvent properties. Rectangular specimens (n=30) were allocated to three groups, according to the concentration of FMA substituted into the monomer component of a heat-polymerized acrylic resin (Lucitone 550), as follows: 0% (control), 10% and 20% (v/v). Acrylic resin mixed with concentrations of 25% or more did not reach the dough stage and was not viable. The surface roughness and flexural strength of the specimens were tested. Variables were analyzed by ANOVA and Tukey's test (a=0.05). Immersion in FMA produced negligible swelling, and MMA produced obvious swelling and dissolution of the specimens. Surface roughness at concentrations of 0%, 10% and 20% were: 0.25 ± 0.04, 0.24 ± 0.04, 0.22 ± 0.03 mm (F=1.78; p=0.189, not significant). Significant differences were found for flexural strength (F=15.92; p<0.001) and modulus of elasticity (F=7.67; p=0.002), with the following results: 96 ± 6, 82 ± 5, 84 ± 6 MPa, and 2,717 ± 79, 2,558 ± 128, 2574 ± 87 MPa, respectively. The solvent properties of FMA against acrylic resin are weak, which would explain why concentrations over 20% were not viable. Surface changes were not detected after the incorporation of FMA in the denture base acrylic resin tested. The addition of FMA into denture base resin may lower the flexural strength and modulus of elasticity, regardless of the tested concentration.

Effect of repeated disinfections by microwave energy on the physical and mechanical properties of denture base acrylic resins

The present study evaluated the effect of repeated simulated microwave disinfection on physical and mechanical properties of Clássico, Onda-Cryl and QC-20 denture base acrylic resins. Aluminum patterns were included in metallic or plastic flasks with dental stone following the traditional packing method. The powder/liquid mixing ratio was established according to the manufacturer's instructions. After water-bath polymerization at 74ºC for 9 h, boiling water for 20 min or microwave energy at 900 W for 10 min, the specimens were deflasked after flask cooling and finished. Each specimen was immersed in 150 mL of distilled water and underwent 5 disinfection cycles in a microwave oven set at 650 W for 3 min. Non-disinfected and disinfected specimens were subjected to the following tets: Knoop hardness test was performed with 25 g load for 10 s, impact strength test was done using the Charpy system with 40 kpcm, and 3-point bending test (flexural strength) was performed at a crosshead speed of 0.5 mm/min until fracture. Data were analyzed statistically by ANOVA and Tukey's test (α= 0.05%). Repeated simulated microwave disinfections decreased the Knoop hardness of Clássico and Onda-Cryl resins and had no effect on the impact strength of QC-20. The flexural strength was similar for all tested resins.

Can previous acid etching increase the bond strength of a self-etching primer adhesive to enamel?

Because a greater research effort has been directed to analyzing the adhesive effectiveness of self etch primers to dentin, the aim of this study was to evaluate, by microtensile testing, the bond strength to enamel of a composite resin combined with a conventional adhesive system or with a self-etching primer adhesive, used according to its original prescription or used with previous acid etching. Thirty bovine teeth were divided into 3 groups with 10 teeth each (n= 10). In one of the groups, a self-etching primer (Clearfil SE Bond - Kuraray) was applied in accordance with the manufacturer's instructions and, in the other, it was applied after previous acid etching. In the third group, a conventional adhesive system (Scotchbond Multipurpose Plus - 3M-ESPE) was applied in accordance with the manufacturer's instructions. The results obtained by analysis of variance revealed significant differences between the adhesive systems (F = 22.31). The self-etching primer (Clearfil SE Bond) presented lower enamel bond strength values than the conventional adhesive system (Scotchbond Multipurpose Plus) (m = 39.70 ± 7.07 MPa) both when used according to the original prescription (m = 27.81 ± 2.64 MPa) and with previous acid etching (m = 25.08 ± 4.92 MPa).

Temperature of denture base resin under different protocols of microwave irradiation

This in vitro study evaluated the temperature of dentures after different microwave irradiation protocols. Two complete dentures (one maxillary and one mandibular denture) were irradiated separately 4 times for each of the following 5 protocols: dentures immersed in water (G1- 6 min, G2- 3 min); dentures kept dry (G3- 6 min); dentures placed in the steam sterilizer (G4- 6 min, G5- 3 min). The final temperature of the dentures was gauged in a thin and in a thick area of each denture with an infrared thermometer. All groups presented an increase in the resin base temperature. The thin areas of the dentures underwent greater heating than the thick areas. There was no significant difference (p>0.05) between the final mean temperatures of dentures immersed in water for 6 (G1) and 3 min (G2). However, the final mean temperatures recorded in G1 and G2 exceeded 71°C and were significantly higher (<0.001) than the final mean temperatures recorded in the other groups. It may be concluded that denture base resins subjected to microwave irradiation immersed in water may be exposed to deleterious temperatures.

Influência do grau de hidrólise do poli(vinil álcool) nas propriedades físicas de filmes à base de blendas de gelatina e poli(vinil álcool) plastificados com glicerol

O objetivo deste trabalho foi o estudo do efeito do Grau de Hidrólise (GH) do poli(vinil álcool) (PVA) nas propriedades dos filmes à base de blendas de gelatina suína e PVA com dois GH. Os filmes foram produzidos com soluções com 2 g de macromoléculas/100 g de solução, contendo 23,1 g de PVA.100 g-1 de macromoléculas e 25 g de glicerol/100 g de macromoléculas. As propriedades mecânicas e térmicas, cor, opacidade, umidade e solubilidade, além de espectros de infravermelho com transformada de Fourier (FTIR) dos filmes, foram estudadas. As soluções foram analisadas por reometria dinâmica. Os filmes produzidos com o PVA de menor GH foram mais higroscópicos e mais solúveis. Mas o tipo de PVA não afetou a cor, afetando a opacidade e o brilho dos filmes. O PVA com maior GH proporcionou filmes mais resistentes, e o PVA de menor GH produziu filmes mais resistentes à tração, embora menos deformáveis na perfuração. O GH do PVA não afetou a temperatura de transição vítrea dos filmes, determinada na primeira varredura, mas a afetou na segunda varredura. Os resultados das análises de FTIR corroborraram com esses resultados. As propriedades viscoelásticas das soluções não foram afetadas pelo GH do PVA, muito possivelmente por se tratar de soluções diluídas.

Fontes de metionina em rações formuladas com base em aminoácidos totais ou digestíveis para frangas de reposição leves e semipesadas

Foram conduzidos dois experimentos com o objetivo de avaliar a utilização de duas fontes de metionina (em pó ou líquida) em rações formuladas com base em aminoácidos digestíveis ou totais para frangas de reposição leves e semipesadas, nas fases inicial (de 1 a 6 semanas de idade) e de crescimento (de 11 a 16 semanas de idade). Em cada experimento, foram utilizadas 384 aves distribuídas em delineamento inteiramente ao acaso, em esquema fatorial 2 × 2 × 2, composto de duas linhagens (Hy-Line W36 e Hy-Line Brown), duas fontes de metionina (HMTBA - 88% e DL-Metionina - 99%) e dois critérios de formulação da dieta (aminoácidos totais e digestíveis), totalizando oito tratamentos, cada um com seis repetições de oito aves. Foram avaliados os consumos de ração, proteína bruta e energia metabolizável, o peso e ganho de peso corporal e a conversão alimentar das aves. Não houve interação entre os fatores para nenhuma das variáveis estudadas. As aves da linhagem Brown apresentaram maior consumo de ração, peso corporal e ganho de peso. As fontes de metionina e os critérios de formulação de ração não influenciaram as características de desempenho. As aves das linhagens W36 e Brown da linhagem Hy-Line apresentam diferentes perfis de desenvolvimento corporal. A metionina em pó (DL-metionina - 99%) ou a líquida (HMTBA - 88%), assim como os critérios de formulação de ração (aminoácidos totais ou digestíveis) proporcionam desempenho semelhante em aves de reposição.

Desempenho e rendimento de carcaça de frangos de corte alimentados com dietas contendo subprodutos do arroz formuladas com base nos conceitos de proteína bruta e ideal

Foi conduzido um experimento para avaliar a utilização de subprodutos do arroz em dietas formuladas com base nos conceitos de proteína bruta e ideal para frangos de corte de 1 a 42 dias de idade. Foram utilizados 720 pintos machos de 1 dia de idade da linhagem "Hybro", em delineamento inteiramente ao acaso, em esquema fatorial 3 × 2, composto de três dietas (sem subprodutos, farelo de arroz integral e quirera de arroz) e dois conceitos de formulação de rações (proteína bruta e ideal), totalizando seis tratamentos e quatro repetições de 30 aves. O ganho de peso, o consumo de ração e a conversão alimentar foram avaliados aos 21 e 42 dias e as características de carcaça aos 42 dias de idade. As aves alimentadas com dietas formuladas pelo conceito tradicional (baseado na proteína bruta) apresentaram melhor conversão alimentar e menor taxa de deposição de gordura abdominal.

Biodisponibilidade de fontes orgânicas e inorgânicas de zinco em ovinos

Compararam-se os efeitos de diferentes fontes e doses de zinco na dieta de ovinos Santa Inês sobre os níveis de zinco plasmático e de fosfatase alcalina. Foram utilizados 40 cordeiros, recém-desmamados, com média de peso de 18,4kg, distribuídos em 10 tratamentos: 1- dieta basal sem suplementação de zinco; 2- dieta basal + 200mg de Zn/kg de MS na forma de óxido de zinco; 3- dieta basal + 400mg de Zn/kg de MS na forma de óxido de zinco; 4- dieta basal + 600mg de Zn/kg de MS na forma de óxido de zinco; 5- dieta basal + 200mg de Zn/kg de MS na forma de zinco aminoácido; 6- dieta basal + 400mg de Zn/kg de MS na forma de zinco aminoácido; 7- dieta basal + 600mg de Zn/kg de MS na forma de zinco aminoácido; 8- dieta basal + 200mg de Zn/kg de MS na forma de zinco proteinato; 9- dieta basal + 400mg de Zn/kg de MS na forma de zinco proteinato; 10- dieta basal + 600mg de Zn/kg de MS na forma de zinco proteinato. A cada 28 dias, os animais foram pesados e tiveram seu sangue colhido para análise de zinco (Zn), análise de fosfatase alcalina e análises de imunoglobulinas G (IgG) e M (IgM). No final do experimento, foram coletadas amostras de fígado para estudo dos níveis de zinco hepático. Não houve diferença entre tratamentos nos níveis de fosfatase alcalina e de zinco hepático, e no ganho de peso (P>0,05), mas houve diferença (P<0,05) nos níveis de Zn plasmático e nos níveis de IgG e IgM. Baseando-se no acúmulo no fígado, a estimativa da biodisponibilidade de zinco, por intermédio das equações de regressão, mostrou que as fontes orgânicas e inorgânicas de zinco não diferiram entre si.

The role of γ -aminobutyric acid (Gaba) in somatic embryogenesis of Acca sellowiana Berg. (Myrtaceae)

The γ-aminobutyric acid (Gaba) is a non-protein amino acid found in prokaryotes and eukaryotes. Its role in plant development has not been fully established. This study reports a quantification of the levels of endogenous Gaba, as well as investigation of its role in different stages of somatic embryogenesis in Acca sellowiana Berg. (Myrtaceae). Zygotic embryos were used as explants and they were inoculated into the culture medium contained different concentrations of Gaba (0,2, 4, 6, 8 and 10 µM). The highest concentrations of endogenous Gaba were detected between the third and nine days after inoculation, reaching the value of 12.77 µmol.g-1FW. High frequency of somatic embryogenesis was observed in response to 10 µM Gaba. This treatment also resulted in a large number of normal embryos, and the lowest percentage of formation of fused somatic embryos, phenotypic characteristic of most deformed embryos in all treatments. Also, all treatments promoted the formation of the somatic embryos with positive characteristics of development resumption, which however did not originate the seedlings.

Tensile bond strength and SEM analysis of enamel etched with Er:YAG laser and phosphoric acid: a comparative study In vitro

Er:YAG laser has been studied as a potential tool for restorative dentistry due to its ability to selectively remove oral hard tissue with minimal or no thermal damage to the surrounding tissues. The purpose of this study was to evaluate in vitro the tensile bond strength (TBS) of an adhesive/composite resin system to human enamel surfaces treated with 37% phosphoric acid, Er:YAG laser (lambda=2.94 mum) with a total energy of 16 J (80 mJ/pulse, 2Hz, 200 pulses, 250 ms pulse width), and Er:YAG laser followed by phosphoric acid etching. Analysis of the treated surfaces was performed by scanning electron microscopy (SEM) to assess morphological differences among the groups. TBS means (in MPa) were as follows: Er:YAG laser + acid (11.7 MPa) > acid (8.2 MPa) > Er:YAG laser (6.1 MPa), with the group treated with laser+acid being significantly from the other groups (p=0.0006 and p= 0.00019, respectively). The groups treated with acid alone and laser alone were significantly different from each other (p=0.0003). The SEM analysis revealed morphological changes that corroborate the TBS results, suggesting that the differences in TBS means among the groups are related to the different etching patterns produced by each type of surface treatment. The findings of this study indicate that the association between Er:YAG laser and phosphoric acid can be used as a valuable resource to increase bond strength to laser-prepared enamel.