Ferreira, IMPLV, Pinho O, Monteiro D, Faria S, Cruz S, Perreira A, Roque AC, Tavares P.
2010.
Short communication: Effect of kefir grains on proteolysis of major milk proteins, Feb. JOURNAL OF DAIRY SCIENCE. {93}:{27-31}., Number {1}
AbstractThe effect of kefir grains on the proteolysis of major milk proteins in milk kefir and in a culture of kefir grains in pasteurized cheese whey was followed by reverse phase-HPLC analysis. The reduction of kappa-, alpha-, and beta-caseins (CN), alpha-lactalbumin (alpha-LA), and beta-lactoglobulin (beta-LG) contents during 48 and 90 h of incubation of pasteurized milk (100 mL) and respective cheese whey with kefir grains (6 and 12 g) at 20 degrees C was monitored. Significant proteolysis of alpha-LA and kappa-, alpha-, and beta-caseins was observed. The effect of kefir amount (6 and 12 g/100 mL) was significant for alpha-LA and alpha- and beta-CN. alpha-Lactalbumin and beta-CN were more easily hydrolyzed than alpha-CN. No significant reduction was observed with respect to beta-LG concentration for 6 and 12 g of kefir in 100 mL of milk over 48 h, indicating that no significant proteolysis was carried out. Similar results were observed when the experiment was conducted over 90 h. Regarding the cheese whey kefir samples, similar behavior was observed for the proteolysis of alpha-LA and beta-LG: alpha-LA was hydrolyzed between 60 and 90% after 12 h (for 6 and 12 g of kefir) and no significant beta-LG proteolysis occurred. The proteolytic activity of lactic acid bacteria and yeasts in kefir community was evaluated. Kefir milk prepared under normal conditions contained peptides from proteolysis of alpha-LA and kappa-, alpha-, and beta-caseins. Hydrolysis is dependent on the kefir: milk ratio and incubation time. beta-Lactoglobulin is not hydrolyzed even when higher hydrolysis time is used. Kefir grains are not appropriate as adjunct cultures to increase beta-LG digestibility in whey-based or whey-containing foods.
Rodrigues, JEA, Erny GL, Barros AS, Esteves VI, Brandao T, Ferreira AA, Cabrita E, Gil AM.
2010.
Quantification of organic acids in beer by nuclear magnetic resonance (NMR)-based methods, AUG 3 2010. Analytica Chimica Acta. 674:166-175., Number 2
Abstract
Pontes, H, de Pinho PG, Fernandes E, Branco PS, Ferreira LM, Carmo H, Remiao F, Carvalho F, Bastos ML.
2010.
Metabolic interactions between ethanol and MDMA in primary cultured rat hepatocytes, APR 11. TOXICOLOGY. 270:150-157., Number 2-3
Abstract3,4-Methylenedioxymethamphetamine (MDMA; ecstasy), a drug of abuse commonly consumed at rave parties, is often taken in a polydrug abuse scenario, ethanol being one of the most associated drugs. Both MDMA and ethanol are mainly metabolized in the liver with formation of toxic metabolites. Our working hypothesis is that ethanol can modify the metabolism of MDMA through the cytochrome P450 system, and that this effect may be further potentiated by hyperthermia, a well-known consequence of MDMA abuse. To investigate these putative interactions we used primary rat hepatocyte cultures, which were exposed to 300 mM ethanol, 1.6 mM MDMA and the combination of both, at normothermic (36.5 degrees C) and hyperthermic (40.5 degrees C) conditions. After 24 h, the levels of MDA, HMA and HMMA in the cell culture medium were quantified by GC/MS. In addition, we repeated the same experimental design preceded by 1 h incubation with 0.18 mu M ketoconazole or 150 mu M diallyl sulphide (CYP3A and CYP2E1 inhibitors, respectively), to evaluate the putative role of these isoenzymes in the observed effects. The results obtained showed that ethanol exposure increases the formation of some MDMA metabolites such as HMA (1.8 times increase) and MDA (1.5 times increase). This effect was markedly increased under hyperthermic conditions (HMA, MDA and HMMA formation increased 10,6 and 16 times, respectively) and is mediated, at least partially, by CYP3A and CYP2E1. (C) 2010 Elsevier Ireland Ltd. All rights reserved.
Branco, A, Pinheiro C, Fonseca J, Tedim J, Carneiro A, Parola AJ, Freire C, Pina F.
2010.
Solid-State Electrochromic Cells Based on M(salen) -Derived Electroactive Polymer Films, 2010. Electrochemical and Solid State Letters. 13:J114-J118.
AbstractA systematic study of the electrochromic (EC) behavior of electropolymerized poly[M(salen)] films (M = Ni, Cu, and Pd) was performed by spectroelectrochemistry. Color contrast between oxidized and reduced states, stability under square wave potential cycling, coloration efficiency, and switching rate were evaluated. Five polymers were selected to assemble solid-state EC cells in a symmetrical configuration (electrode/poly[M(salen)] film/opaque electrolyte/poly[M(salen)] film/electrode). The best EC performance was found for poly[Pd(3-Mesalen)], poly[1], with 38% of initial diffuse reflectance variation and loss of 50% after 6769 cycles. (C) 2010 The Electrochemical Society. [DOI: 10.1149/1.3457474] All rights reserved.
Ferreira, L.
2010.
Construção de Edíficios Sustentáveis - Contribuição para a definição de um Processo Operativo. Faculdade de Ciências e Tecnologia. (
Amado, Miguel, Ed.)., Lisbon
AbstractA presente Dissertação trata do tema da Construção Sustentável, dando especial enfoque aos Processos Operativos existentes e ao caminho a percorrer para a elaboração de um Processo Operativo enquadrado especificamente dentro dos princípios da Construção Sustentável. Através de uma revisão bibliográfica, procedeu-se ao resumo da evolução dos métodos, técnicas, materiais e fins da construção, desde o início da Humanidade até à actualidade. Apresentou-se, de seguida, o enquadramento geral da realidade nacional e internacional no que respeita à construção de edifícios sustentáveis, tendo em conta as novas premissas para a construção, os novos métodos e os novos processos associados à Construção Sustentável. Neste âmbito, foram analisadas as soluções legislativas vigentes e as propostas analíticas de diversas entidades que se debruçam sobre este tema da Construção Sustentável, enquanto vertente especializada da problemática mais abrangente intrínseca ao aperfeiçoamento e aplicação do conceito de Desenvolvimento Sustentável. Este estudo, integrando uma observação profunda do ciclo de vida do edifício e das especificidades de cada uma das fases que o integram, permitiu cumprir o já referido objectivo primordial da presente Dissertação, contribuindo para o estudo, análise e elaboração de um modelo de investigação que permita avaliar ou elaborar um Processo Operativo para a Construção Sustentável. Desta forma, a presente Dissertação representa um avanço no desenvolvimento de um Processo Operativo para a Construção Sustentável adaptado à realidade da construção em Portugal.
Faria, P.
2010.
Construção sustentável: contributo para o processo de construção na alteração de usos nos edifícios. Faculdade de Ciências e Tecnologia. (
Amado, Miguel, Ed.)., Lisbon
AbstractThe construction industry is currently undergoing a transition phase through which it is trying to
correct the excess of resource consumption occurred during the last two centuries. This industry is
responsible for the consumption of most non-renewable resources, a high consumption of energy and
for and excessive production of waste in all phases of its production process. Thus, the existence of a
building process based on the principle of Sustainable Development capable of reducing the negative
impacts resulting from the current activity is of enormous importance to the society.
One of the present solutions, which is wide spread nearly all over the world, is the process of
rehabilitation of existing buildings with change in use. This process alone can reduce the creation of
demolition waste and resource consumption associated with new construction. When complemented
with a set of sustainable actions, it ensures the reduction of energy consumption and resources
throughout the utilization phase and improves the health of the built environment.
Thus, in order to contribute to the improvement of building´s characteristics and consequently to
society´s quality of life, a building construction process of change in use, based on the principle of
sustainable construction, is presented. This process consists of a set of actions that encompass all
phases of the building’s lifecycle.
Gomes, AQ, Correia DV, Grosso AR, Lança T, Ferreira C, Lacerda JF, Barata JT, da Silva MG, Silva-santos B.
2010.
Identification of a panel of ten cell surface protein antigens associated with immunotargeting of leukemias and lymphomas by peripheral blood γδT cells. Haematologica. 95:1397–1404., Number 8
AbstractBACKGROUND:
Vgamma9Vdelta2 T lymphocytes are regarded as promising mediators of cancer immunotherapy due to their capacity to eliminate multiple experimental tumors, particularly within those of hematopoietic origin. However, Vgamma9Vdelta2 T-cell based lymphoma clinical trials have suffered from the lack of biomarkers that can be used as prognostic of therapeutic success.
DESIGN AND METHODS:
We have conducted a comprehensive study of gene expression in acute lymphoblastic leukemias and non-Hodgkin's lymphomas, aimed at identifying markers of susceptibility versus resistance to Vgamma9Vdelta2 T cell-mediated cytotoxicity. We employed cDNA microarrays and quantitative real-time PCR to screen 20 leukemia and lymphoma cell lines, and 23 primary hematopoietic tumor samples. These data were analyzed using state-of-the-art bioinformatics, and gene expression patterns were correlated with susceptibility to Vgamma9Vdelta2 T cell mediated cytolysis in vitro.
RESULTS:
We identified a panel of 10 genes encoding cell surface proteins that were statistically differentially expressed between "gammadelta-susceptible" and "gammadelta-resistant" hematopoietic tumors. Within this panel, 3 genes (ULBP1, TFR2 and IFITM1) were associated with increased susceptibility to Vgamma9Vdelta2 T-cell cytotoxicity, whereas the other 7 (CLEC2D, NRP2, SELL, PKD2, KCNK12, ITGA6 and SLAMF1) were enriched in resistant tumors. Furthermore, some of these candidates displayed a striking variance of expression among primary follicular lymphomas and T-cell acute lymphoblastic leukemias.
CONCLUSIONS:
Our results suggest that hematopoietic tumors display a highly variable repertoire of surface proteins that can impact on Vgamma9Vdelta2 cell-mediated immunotargeting. The prognostic value of the proposed markers can now be evaluated in upcoming Vgamma9Vdelta2 T cell-based lymphoma/leukemia clinical trials.
Morgado, L, Fernandes AP, Londer YY, Bruix M, Salgueiro CA.
2010.
One simple step in the identification of the cofactors signals, one giant leap for the solution structure determination of multiheme proteins. Biochemical and Biophysical Research Communications. 393(3):466-470.
AbstractMultiheme proteins play major roles in various biological systems. Structural information on these systems in solution is crucial to understand their functional mechanisms. However, the presence of numerous proton-containing groups in the heme cofactors and the magnetic properties of the heme iron, in particular in the oxidised state, complicates significantly the assignment of the NMR signals. Consequently, the multiheme proteins superfamily is extremely under-represented in structural databases, which constitutes a severe bottleneck in the elucidation of their structural–functional relationships. In this work, we present a strategy that simplifies the assignment of the NMR signals in multiheme proteins and, concomitantly, their solution structure determination, using the triheme cytochrome PpcA from the bacterium Geobacter sulfurreducens as a model. Cost-effective isotopic labeling was used to double label (13C/15N) the protein in its polypeptide chain, with the correct folding and heme post-translational modifications. The combined analysis of 1H–13C HSQC NMR spectra obtained for labeled and unlabeled samples of PpcA allowed a straight discrimination between the heme cofactors and the polypeptide chain signals and their confident assignment. The results presented here will be the foundations to assist solution structure determination of multiheme proteins, which are still very scarce in the literature.
Catarino, T, Pessanha M, Candia ADG, Gouveia Z, Fernandes AP, Pokkuluri PR, Murgida D, Marti MA, Todorovic S, Salgueiro CA.
2010.
Probing the Chemotaxis Periplasmic Sensor Domains from Geobacter sulfurreducens by Combined Resonance Raman and Molecular Dynamic Approaches: NO and CO Sensing. The Journal of Physical Chemistry B. 114 (34):11251-11260.
AbstractThe periplasmic sensor domains encoded by genes gsu0582 and gsu0935 are part of methyl accepting chemotaxis proteins in the bacterium Geobacter sulfurreducens (Gs). The sensor domains of these proteins contain a heme-c prosthetic group and a PAS-like fold as revealed by their crystal structures. Biophysical studies of the two domains showed that nitric oxide (NO) binds to the heme in both the ferric and ferrous forms, whereas carbon monoxide (CO) binds only to the reduced form. In order to address these exogenous molecules as possible physiological ligands, binding studies and resonance Raman (RR) spectroscopic characterization of the respective CO and NO adducts were performed in this work. In the absence of exogenous ligands, typical RR frequencies of five-coordinated (5c) high-spin and six-coordinated (6c) low-spin species were observed in the oxidized form. In the reduced state, only frequencies corresponding to the latter were detected. In both sensors, CO binding yields 6c low-spin adducts by replacing the endogenous distal ligand. The binding of NO by the two proteins causes partial disruption of the proximal Fe-His bond, as revealed by the RR fingerprint features of 5cFe-NO and 6cNO-Fe-His species. The measured CO and NO dissociation constants of ferrous GSU0582 and GSU0935 sensors reveal that both proteins have high and similar affinity toward these molecules (Kd ≈ 0.04−0.08 μM). On the contrary, in the ferric form, sensor GSU0582 showed a much higher affinity for NO (Kd ≈ 0.3 μM for GSU0582 versus 17 μM for GSU0935). Molecular dynamics calculations revealed a more open heme pocket in GSU0935, which could account for the different affinities for NO. Taken together, spectroscopic data and MD calculations revealed subtle differences in the binding properties and structural features of formed CO and NO adducts, but also indicated a possibility that a (5c) high-spin/(6c) low-spin redox-linked equilibrium could drive the physiological sensing of Gs cells.
Ferreira, IMPLV, Pinho O, Monteiro D, Faria S, Cruz S, Perreira A, Roque ACA, Tavares P.
2010.
Short communication: effect of kefir grains on proteolysis of major milk proteins. Journal of Dairy Science. 93:27–31., Number 1
AbstractThe effect of kefir grains on the proteolysis of major milk proteins in milk kefir and in a culture of kefir grains in pasteurized cheese whey was followed by reverse {phase-HPLC} analysis. The reduction of kappa-, alpha-, and beta-caseins {(CN)}, alpha-lactalbumin {(alpha-LA)}, and beta-lactoglobulin {(beta-LG)} contents during 48 and 90 h of incubation of pasteurized milk {(100mL)} and respective cheese whey with kefir grains (6 and 12 g) at 20 degrees C was monitored. Significant proteolysis of {alpha-LA} and kappa-, alpha-, and beta-caseins was observed. The effect of kefir amount (6 and 12 {g/100mL)} was significant for {alpha-LA} and alpha- and {beta-CN.} {alpha-Lactalbumin} and {beta-CN} were more easily hydrolyzed than {alpha-CN.} No significant reduction was observed with respect to {beta-LG} concentration for 6 and 12 g of kefir in {100mL} of milk over 48 h, indicating that no significant proteolysis was carried out. Similar results were observed when the experiment was conducted over 90 h. Regarding the cheese whey kefir samples, similar behavior was observed for the proteolysis of {alpha-LA} and {beta-LG:} {alpha-LA} was hydrolyzed between 60 and 90% after 12h (for 6 and 12 g of kefir) and no significant {beta-LG} proteolysis occurred. The proteolytic activity of lactic acid bacteria and yeasts in kefir community was evaluated. Kefir milk prepared under normal conditions contained peptides from proteolysis of {alpha-LA} and kappa-, alpha-, and beta-caseins. Hydrolysis is dependent on the kefir:milk ratio and incubation time. {beta-Lactoglobulin} is not hydrolyzed even when higher hydrolysis time is used. Kefir grains are not appropriate as adjunct cultures to increase {beta-LG} digestibility in whey-based or whey-containing foods.