From Rational Design of Drug Crystals to Understanding of Nucleic Acid Structures: Lamivudine Duplex

A DNA-like duplex of nucleosides is probable to exist even without the 5`-phosphate groups needed to assemble the chain backbone. However, double-stranded helical structures of nucleosides are unknown. Here, we report a duplex of nucleoside analogs that is spontaneously assembled due to stacking of the neutral and protonated molecules of lamivudine, a nucleoside reverse transcriptase inhibitor (NTRI) widely used in anti-HIV drug combinatory medication. The left-handed lamivudine duplex has features similar to those of i-motif DNA, as the face-to-face base stacking and the helix rise per base pair. Furthermore, the protonation pattern on alternate bases expected for it DNA-like duplex stabilized by pairing of neutral and protonated cytosine fragments was observed for the first time in the lamivudine double-stranded helix. This structure demonstrates that hydrogen bonds can substitute for covalent phosphodiester linkage in the stabilization of the duplex backbone. This interesting example of spontaneous molecular self-organization indicates that the 5`-phosphate group could not be a requirement for duplex assembly.

DNA strand breaks and base modifications induced by cholesterol hydroperoxides

Cholesterol (Ch) can be oxidized by reactive oxygen species, forming oxidized products such as Ch hydroperoxides (ChOOH). These hydroperoxides can disseminate the peroxidative stress to other cell compartments. In this work, the ability of ChOOH to induce strand breaks and/or base modifications in a plasmid DNA model was evaluated. In addition, HPLC/MS/MS analyses were performed to investigate the formation of 8-oxo-7,8-dihydro-2`-deoxyguanosine (8-oxodGuo) after the incubation of 2`-deoxyguanosine (dGuo) with ChOOH and Cu(2+). In the presence of copper ions, ChOOH induced DNA strand breaks in time and concentration-dependent manners. Purine and pyrimidine base modifications were also observed, as assessed respectively by the treatment with Fpg and Endo III repair enzymes. The detection of 8-oxodGuo by HPLC/MS/MS is in agreement with the dGuo oxidation in plasmid DNA. ChOOH-derived DNA damage adds further support to the role of lipid peroxidation in inducing DNA modifications and mutation.

Double-strand DNA cleavage induced by oxindole-Schiff base copper(II) complexes with potential antitumor activity

Some oxindole-Schiff base copper(II) complexes have already shown potential antitumor activity towards different cells, inducing apoptosis in a process modulated by the ligand, and having nuclei and mitochondria as main targets. Here, three novel copper(II) complexes with analogous ligands were isolated and characterized by spectroscopic techniques, having their reactivity compared to the so far most active complex in this class. Cytotoxicity experiments carried out toward human neuroblastoma SH-SY5Y cells confirmed its proapoptosis property. DNA cleavage studies were then performed in the presence of these complexes, in order to verify the influence of ligand structural features in its nuclease activity. All of them were able to cause double-strand DNA scissions, giving rise to nicked circular Form II and linear Form III species, in the presence of hydrogen peroxide. Additionally, DNA Form II was also detected in the absence of peroxide when the most active complex, [Cu(isaepy)(2)](2+) 1, was used. In an effort to better elucidate their interactions with DNA, solutions of the different complexes titrated with DNA had their absorption spectra monitored. An absorbance hyperchromism observed at 260 nm pointed to the intercalation of these complexes into the DNA structure. Further, investigations of 2-deoxy-D-ribose (DR) oxidation catalyzed by each of those complexes, using 2-thiobarbituric acid reactive species (TBARS) method, and detection of reactive oxygen species (ROS) formation by spin-trapping EPR, suggested that their mechanism of action in performing efficiently DNA cleavage occurs preferentially, but not only by oxidative pathways. (C) 2007 Elsevier Inc. All rights reserved.

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.

Digestibilidade e balanço de nitrogênio em ovinos alimentados à base de dietas com elevado teor de concentrado e níveis crescentes de polpa cítrica peletizada

O experimento foi realizado com o objetivo de determinar os coeficientes de digestibilidade, a ingestão de nutrientes e o balanço de nitrogênio de dietas com 20% de silagem de milho e 80% de concentrado e 0, 25, 40 e 55% de polpa cítrica na matéria seca total, substituindo o milho em grão. em delineamento de blocos ao acaso, foram utilizados oito cordeiros mestiços, com nove meses de idade e 34 kg de peso, mantidos em gaiolas individuais com dispositivo para separação de fezes e urina. O período de coleta de alimento, sobras, fezes e urina foi de seis dias e a quantidade de alimento oferecido correspondeu à média do consumido nos três últimos dias do período de adaptação. O experimento foi desenvolvido em duas fases, utilizando-se os mesmos animais, que foram distribuídos em blocos pelo peso. Os resultados foram analisados por regressão polinomial. Houve aumento linear nos coeficientes de digestibilidade aparente da proteína bruta e da fibra em detergente ácido, com o aumento da participação da polpa cítrica na dieta. Os demais coeficientes de digestibilidade não diferiram entre si e foram, respectivamente: 71,8; 73,3; 77,2; 63,5; e 72,2%; para matéria seca, matéria orgânica, extrato etéreo, fibra em detergente neutro e hemicelulose. O teor médio de nutrientes digestíveis totais foi de 72,39%. Houve aumento linear da ingestão de matéria seca e de nutrientes digestíveis totais por dia e por peso metabólico, com o aumento de polpa cítrica na dieta. O mesmo ocorreu com a quantidade de nitrogênio ingerido, absorvido, urinário e retido. O aumento da participação da polpa cítrica peletizada até 55% da matéria seca da dieta, substituindo o grão de milho, implicou em melhor aproveitamento da fibra e eficiência no metabolismo do nitrogênio.

Digestibilidade e degradabilidade de rações à base de milho desintegrado com palha e sabugo em diferentes graus de moagem

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

Formulação de rações para poedeiras com base em aminoácidos totais e digestíveis utilizando diferentes estimativas da composição de aminoácidos em alimentos

Foram utilizadas poedeiras comerciais com 27 semanas de idade, distribuídas em delineamento inteiramente ao acaso, em esquema fatorial 3 x 3, com três repetições de seis aves por tratamento. Os fatores consistiram de três métodos de estimativa da composição de aminoácidos em ingredientes (tabelas brasileiras, equações de predição e fator para correção de aminoácido em função do teor de proteína do ingrediente) e três recomendações de aminoácidos, sendo duas de aminoácidos digestíveis e uma de aminoácidos totais. Os métodos de estimativa da composição de aminoácidos nos ingredientes afetaram apenas a conversão alimentar e a espessura de casca, que apresentaram os melhores resultados com a utilização das tabelas brasileiras. Embora as recomendações de aminoácidos tenham determinado diferenças em todos os parâmetros de desempenho, não afetaram a qualidade dos ovos. O desempenho das aves foi prejudicado pelos níveis de aminoácidos digestíveis, entretanto, ambas as recomendações promoveram desempenho semelhante e inferior ao de aminoácidos totais. O pior desempenho das aves alimentadas com as rações formuladas com aminoácidos digestíveis pode ser atribuído à deficiência em nitrogênio para a síntese de aminoácidos não-essenciais, visto que o nível protéico foi reduzido (12,5% PB), ou ainda à deficiência nos aminoácidos arginina, histidina, isoleucina, leucina e valina, cujos requerimentos mínimos não foram considerados na formulação das rações.

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.

Composição em ácidos graxos e qualidade da carne de tourinhos Nelore e Canchim alimentados com dietas à base de cana-de-açúcar e dois níveis de concentrado

O objetivo neste trabalho foi avaliar a composição de ácidos graxos e a qualidade do contrafilé (músculo Longissimus lumborum) de tourinhos das raças Nelore e Canchim. Os animais foram terminados em confinamento e alimentados com dietas contendo cana-de-açúcar e dois níveis de concentrado (40 e 60% na matéria seca). Os concentrados foram compostos de grãos de girassol, milho, farelo de soja, levedura seca de cana-de-açúcar, uréia e núcleo mineral. O delineamento experimental foi o inteiramente ao acaso, em esquema fatorial 2 × 2 (grupo genético × nível de concentrado). Não foram observadas diferenças nos teores de umidade, proteína e extrato etéreo da carne. Os animais da raça Nelore apresentaram maiores concentrações de ácido linoléico conjugado (0,52%), ácidos graxos insaturados (46,82%) e também relações mais elevadas de ácidos graxos insaturados:saturados (1,02) e monoinsaturados:saturados (0,86) em comparação aos tourinhos da raça Canchim. Os tourinhos da raça Canchim apresentaram maior intensidade das cores vermelha e amarela no contrafilé e maior luminosidade da gordura de cobertura. Houve interação para força de cisalhamento, que foi menor nos tourinhos Nelore alimentados com 40% de concentrado. Tourinhos da raça Nelore apresentam carne com melhor composição de ácidos graxos na gordura intramuscular do ponto de vista da saúde humana.

Lesão na base e concentrações de ácido indolbutírico no enraizamento de estacas apicais de figueira ‘Roxo de Valinhos’

This work aimed to increase the rhizogenic potential of cuttings collected from the apical portion of the branches of fig trees, performing injuries and treating the cuttings with indolbutyric acid (IBA). Apical cuttings of 'Roxo de Valinhos' fig tree were collected in July. The cuttings were standardized with 20 cm in length and basal diameter of 10 mm. The cuttings received or not incisions at the basis (parallel cuts of 2 cm) and immersed in IBA at 0, 1000, 2000 and 3000 mg L(-1) for 10 seconds. The cuttings were buried (3/4 of the length) in moistened sand, inside a screen house (50% of light). After 60 days it was found that treatment with IBA benefits in the development of apical cuttings, and the concentration that achieved the best results was 2000 mg L(-1); the use of injury at the base of the cuttings helps rooting.

Preparação de resinas de poliuretana à base de óleo de mamona e dietanolamina

The aim of this work was the preparation of polyols from reactions between castor oil and dietanolamine to increase the hydroxyl content and the network degree in the products to application in electronic devices. The polyols and the mixtures obtained were characterized by nuclear magnetic ressonance. Castor oil (CO) is a natural triglyceride - based polyol possessing hydroxyl groups, which allow several reactions that produce many different products. Among them are the polyurethanes (PU), which have been considered an ideal product for the covering of electricelectronic circuits, due to their excellent electrical, shock-absorbing, solvents resistance and hydrolytic stability properties. About 90% of the fatty acids present in the castor oil are ricinoleic acid (12-hydroxyoleic acid), while the remaining 10% correspond to non-hydroxylated fatty acids, mainly linoleic and oleic acids. The chemical analysis of castor oil indicates a hydroxyl number of 2.7. In this work, a polyol was obtained by the reaction of the CO with diethanolamine (DEA), in order to elevate the hydroxyl value from 160 to 230 or to 280 mgKOH/g, and characterized by nuclear magnetic resonance (NMR) 1H and 13C (Mercury 200). The polyadition of the resulting polyol with isophorone diisocianate (IPDI) was carried out at 60°C, and the reaction kinetics was followed by rheological measurements in a Haake RS150 rheometer. The electrical properties were determined in a HP LCR Meter 4262A, at 1.0 Hz and 10.0 KHz. The chemical analysis showed that the polyols obtained presented hydroxyl number from 230 to 280 mgKOH/g. The polyadition reaction with IPDI produced polyurethane resins with the following properties: hardness in the range from 45 shore A to 65 shore D (ASTM D2240); a dielectric constant of 3.0, at 25°C (ASTM D150). Those results indicate that the obtained resins present compatible properties to the similar products of fossil origin, which are used nowadays for covering electric-electronic circuits. Therefore, the PUs from castor oil can be considered as alternative materials of renewable source, free from the highly harmful petroleum - derived solvents

A contextualização e a interdisplinaridade no ensino de Química: uma analise de livros didáticos ACIDO-BASE e das propostas pedagógicas realizadas pelos docentes diante da temática

Nowadays, chemistry contents taught in high school continue to be presented in a fragmented and decontextualized manner by the teachers and the textbooks. Even though it is known that contextualization and interdisciplinary exchange play an important role in the process of Chemistry teaching/learning. Therefore, the present study aims at enlightening the importance of these methodological foundations in the learning of chemistry. The data acquisition about the subject Contextualization and Interdisciplinary Exchange involved in Chemistry Teaching was developed through bibliographical researches on chemistry textbooks, which focused on the analysis of the topics acid and base , since it is a theme studied throughout all three years of high school. The present study also developed questionnaires which were applied to analyze to what extent chemistry teachers are working in a contextualized and interdisciplinary manner throughout the process of Chemistry teaching/learning. The results obtained in the researches show that a contextualized and interdisciplinary teaching contributes to a more meaningful acquisition of chemistry knowledge, in a dynamic and interactive way, but there are still many roadblocks towards the achievement of this kind of Chemistry teaching/learning process

Obtenção de dispersões de complexos polieletrolíticos à base de quitosana e poli(ácido metacrílico) e análise de adsorção de albumina bovina sérica

Dispersions composed of polyelectrolyte complexes based on chitosan and poly(methacrylic acid), PMAA, were obtained by the dropping method and template polymerization. The effect of molecular weight of PMAA and ionic strength on the formation of chitosan/poly(methacrylic acid), CS/PMAA, complexes was evaluated using the dropping method. The increase in molecular weight of PMAA inhibited the formation of insoluble complexes, while the increase in ionic strength first favored the formation of the complex followed by inhibiting it at higher concentrations. The polyelectrolyte complexation was strongly dependent on macromolecular dimensions, both in terms of molecular weight and of coil expansion/contraction driven by polyelectrolyte effect. The resultant particles from dropping method and template polymerization were characterized as having regions with different charge densities: chitosan predominating in the core and poly(methacrylic acid) at the surface, the particles being negatively charged, as a consequence. Albumin was adsorbed on templatepolymerized CS/PMAA complexes (after crosslinking with glutardialdehyde) and pH was controlled in order to obtain two conditions: (i) adsorption of positively charged albumin, and (ii) adsorption of albumin at its isoelectric point. Adsorption isotherms and zeta potential measurements showed that albumin adsorption was controlled by hydrogen bonding/van der Waals interactions and that brushlike structures may enhance adsorption of albumin on these particles

Redução da proteína da dieta com base no conceito de proteína ideal para tilápias-do-nilo (Oreochromis niloticus) criadas em tanques-rede

Avaliou-se o desempenho de tilápias-do-nilo (Oreochromis niloticus) criadas em tanques-rede e alimentadas com dietas contendo 27,0 (controle); 25,2; 24,3 e 22,7% de proteína digestível. Aminoácidos cristalinos (L-lisina, DL-metionina e L-treonina) foram adicionados à dieta considerando o conceito de proteína ideal e simulando o perfil de aminoácidos da dieta controle. Os peixes (34,63±19 g) foram alimentados manualmente com dietas isoenergéticas (3.075 kcal de energia digestível/kg de dieta) até saciedade aparente, três vezes ao dia, durante 91 dias. Utilizou-se um delineamento inteiramente casualizado, com quatro tratamentos, três repetições e 25 peixes/unidade experimental. Não foram observados efeitos dos níveis de proteína digestível sobre o ganho de peso, a conversão alimentar, a taxa de eficiência protéica, o peso da carcaça eviscerada, o rendimento de carcaça, o peso e o rendimento de filé, a sobrevivência e o hematócrito. Houve efeito quadrático dos níveis de proteína digestível sobre o consumo; o maior valor foi estimado para a dieta contendo 24,41% de proteína digestível e excreção de nitrogênio, na qual o melhor resultado estimado foi obtido com peixes que receberam a dieta contendo 24,92% de PD. Com a redução nos níveis de proteína digestível, observou-se aumento linear na retenção de nitrogênio. É possível reduzir o nível de proteína digestível, de 27 (29,1% de PB) para 24,3% (26,6% de PB), em dietas para tilápias-do-nilo criadas em tanques-rede. Essa redução deve ser feita por meio da suplementação de aminoácidos (com base no conceito de proteína ideal), considerando o desempenho e o custo da dieta/kg ganho em filé.