On the interaction of bovine serum albumin with ionic surfactants: Temperature induced EPR changes of a maleimide nitroxide reflect local protein dynamics and probe solvent accessibility

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Características químicas de substratos utilizados na produção de mudas de cacaueiros

O objetivo deste trabalho foi realizar monitoramento químico de seis substratos utilizados na produção de mudas de cacaueiros. Os substratos receberam 10 volumes (100mL, cada) de água destilada, e no lixiviado coletado foram determinados o pH, a condutividade elétrica (CE) e os teores de sódio (Na) e potássio (K) em solução. Os valores de pH variaram de 5,0 a 8,0 em Plantmax® e composto do tegumento da amêndoa do cacau (CTAC), respectivamente. Verificou-se que, a partir do primeiro volume lixiviado, a CE de todos os substratos foi reduzida. Foram encontradas correlações positivas para a CE em relação ao K e Na da fibra de coco (FC) (r=0,83 e 0,72 P<0,05) e CTAC (r=0,98 e 0,99 P<0,05). Os resultados indicam que a FC e o CTAC deveriam ser usados após pré-lavagens. Verificou-se que, embora a FC apresente na primeira lixiviação valor de CE superior aos demais substratos, o valor ideal da 2,0 dSm-1 é alcançado na segunda lixiviação. Os teores de K e Na são um indicativo do valor da CE em FC e CTAC.

Substratos e estaquia na produção de mudas de cacaueiro

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Interaction of giant extracellular Glossoscolex paulistus hemoglobin (HbGp) with ionic surfactants: A MALDI-TOF-MS study

The giant extracellular hemoglobin of Glossoscolex paulistus (HbGp) is constituted by approximately 144 subunits containing heme groups with molecular masses in the range of 16-19 kDa forming a monomer (d) and a trimer (abc), and around 36 non-heme structures, named linkers (L). Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF-MS) analysis was performed recently, to obtain directly information on the molecular masses of the different subunits from HbGp in the oxy-form. This technique demonstrated structural similarity between HbGp and the widely studied hemoglobin of Lumbricus terrestris (HbLt). Indeed, two major isoforms (d(1) and d(2)) of identical proportions with masses of 16,355+/-25 and 16,428+/-24 Da, respectively, and two minor isoforms (d(3) and d(4)) with masses around 16.6 kDa were detected for monomer d of HbGp. In the present work, the effects of anionic sodium dodecyl sulfate (SDS) and cationic cethyltrimethyl ammonium chloride (CTAC) on the oligomeric structure of HbGp have been studied by MALDI-TOF-MS in order to evaluate the interaction between ionic surfactants and HbGp. The data obtained with this technique show an effective interaction of cationic surfactant CTAC with the two isoforms of monomer d, d(1) and d(2), both in the whole protein as well as in the pure isolated monomer. The results show that up to 10 molecules of CTAC are bound to each isoform of the monomer. Differently, the mass spectra obtained for SDS-HbGp system showed that the addition of the anionic surfactant SDS does not originate any mass increment of the monomeric subunits, indicating that SDS-HbGp interaction is, probably, significantly less effective as compared to CTAC-HbGp one. The acid pI of the protein around 5.5 is, probably, responsible for this behavior. The results of this work suggest also some interaction of both surfactants with linker chains as well as with trimers, as judged from observed mass increments. Our data are consistent with a recent spectroscopic study showing a strong interaction between CTAC and HbGp at physiological pH [P.S.Santiago, et al, Biochim. Biophys. Acta. 1770 (2007) 506-517.]. (C) 2007 Elsevier B.V. All rights reserved.

Effect of some surfactants on the chemiluminescent reactions of bis(2,4,6-trichlorophenyl)oxalate and bis(2-nitrophenyl)oxalate with hydrogen peroxide

The chemiluminescent reactions of bis(2,4,6-trichlorophenyl)oxalate (TCPO) and bis(2-nitrophenyl)oxalate (2-NPO) with hydrogen peroxide in acetonitrile/water micellar systems (anionic, cationic, and non-ionic) and gamma-cyclodextrin were studied in the presence of fluoranthene or 9,10-diphenylanthracene, imidazole, and two buffer solutions, HTRIS+/TRIS and H2PO4-/HPO42-. The relative chemiluminenscence (CL) intensity is higher in the presence of the cationic (DDAB, CTAC, DODAC, and OTAC), anionic (SDS), and non-ionic (Tween 80) surfactants. In the presence of some non-ionic surfactants (Brij 35, Brij 76, and Tween 20), the CL intensity was partially quenched compared with the reaction with no surfactant. The sensitivity for hydrogen peroxide determination in the range 0.01 x 10(-4) to 1.0 x 10(-4) mol L-1, considering the slope of the calibration curves (maximum peak height of CL vs. concentration), improved with the introduction of DDAH, CTAB, and SDS in HTRIS+/TRIS buffer.

On the interaction of bovine serum albumin with ionic surfactants: Temperature induced EPR changes of a maleimide nitroxide reflect local protein dynamics and probe solvent accessibility

The interaction of bovine serum albumin (BSA) with the ionic surfactants sodium dodecylsulfate (SDS, anionic), cetyltrimethylammonium chloride (CTAC, cationic) and N-hexadecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate (HPS, zwitterionic) was studied by electron paramagnetic resonance (EPR) spectroscopy of spin label covalently bound to the single free thiol group of the protein. EPR spectra simulation allows to monitor the protein dynamics at the labeling site and to estimate the changes in standard Gibbs free energy, enthalpy and entropy for transferring the nitroxide side chain from the more motionally restricted to the less restricted component. Whereas SDS and CTAC showed similar increases in the dynamics of the protein backbone for all measured concentrations. HPS presented a smaller effect at concentrations above 1.5 mM. At 10 mM of surfactants and 0.15 mM BSA, the standard Gibbs free energy change was consistent with protein backbone conformations more expanded and exposed to the solvent as compared to the native protein, but with a less pronounced effect for HPS. In the presence of the surfactants, the enthalpy change, related to the energy required to dissociate the nitroxide side chain from the protein, was greater, suggesting a lower water activity. The nitroxide side chain also detected a higher viscosity environment in the vicinity of the paramagnetic probe induced by the addition of the surfactants. The results suggest that the surfactant-BSA interaction, at higher surfactant concentration, is affected by the affinities of the surfactant to its own micelles and micelle-like aggregates. Complementary DLS data suggests that the temperature induced changes monitored by the nitroxide probe reflects local changes in the vicinity of the single thiol group of Cys-34 BSA residue. (C) 2011 Elsevier B.V. All rights reserved.

A importância do sistema de lições aprendidas para a instituição militar

As Lições Aprendidas (LL), permitem a uma organização aprender com a experiência e desta forma melhorar o seu desempenho. No Estado-Maior General das Forças Armadas (EMGFA), esta capacidade, encontra-se em fase de implementação, e por este motivo o trabalho realizado, teve como finalidade formular subsídios para o seu desenvolvimento. Através de uma estratégia de pesquisa qualitativa, foi analisado o modelo implementado no Joint Analysis and Lessons Learned Centre (JALLC) e o modelo que se encontra em implementação no Centro de Treino Avaliação e Certificação (CTAC), com vista a identificar as linhas de ação para o desenvolvimento da capacidade. A NATO tem a sua capacidade sustentada no JALLC, que assegura resposta a requisitos de análise, promove a partilha de informações e auxilia no desenvolvimento de LL. No EMGFA o CTAC tem a atribuição de validar as lições identificadas, estabelecer e promover a implementação das lições aprendidas e conduzir estudos de investigação e análise. Do nosso estudo, concluímos que o CTAC está a desenvolver a capacidade de LL de acordo com a doutrina pela NATO, no entanto o seu âmbito de atuação carece de ser alargado, para edificar a capacidade de LL no EMGFA. Abstract: The Lessons Learned (LL), allow an organization to learn from experience and thus improve their performance. In the Portuguese General Staff of the Armed Forces (EMGFA), this capability is being implemented, and for this reason this study, aimed to formulate subsidies for its development. Through a qualitative research strategy, both model the Joint Analysis and Lessons Learned Centre(JALLC) and the the Training Center Assessment and Certification (CTAC) was analysed, to identify the lines of action for the development of capacity in EMGFA. NATO supports their capability in JALLC, which ensures response to requirements analysis, promotes the sharing of information and assists in the development of lessons learned. CTAC in EMGFA has the authority to validate the identified lessons, establish and promote the implementation of lessons learned and conduct research and analysis studies. From our study, we conclude that CTAC is developing LL capacity according to NATO doctrine, however their performance scope needs to be extended to build the capacity in LL EMGFA.