Exploiting the versatility of human cytochrome P450 enzymes: The promise of blue roses from biotechnology

The cytochrome P450 (P450) enzymes involved in drug metabolism are among the most versatile biological catalysts known. A small number of discrete forms of human P450 are capable of catalyzing the monooxygenation of a practically unlimited variety of xenobiotic substrates, with each enzyme showing a more or less wide and overlapping substrate range. This versatility makes P450s ideally suited as starting materials for engineering designer catalysts for industrial applications. In the course of heterologous expression of P450s in bacteria, we observed the unexpected formation of blue pigments. Although this was initially assumed to be an artifact, subsequent work led to the discovery of a new function of P450s in intermediary metabolism and toxicology, new screens for protein engineering, and potential applications in the dye and horticulture industries.

The blood volumes of the primary and secondary circulatory system in the Atlantic cod Gadus morhua L., using plasma bound Evans Blue and compartmental analysis

The volume of the primary (PCS) and secondary (SCS) circulatory system in the Atlantic cod Gadus morhua was determined using a modified dye dilution technique. Cod (N=10) were chronically cannulated in the second afferent branchial artery with PE-50 tubing. Evans Blue dye was bound to harvested fish plasma at a concentration of 1 mg dye ml(-1) plasma, and injected at a concentration of 1 mg kg(-1) body mass. Serial sampling from the cannula produced a dye dilution curve, which could be described by a double exponential decay equation. Curve analysis enabled the calculation of the primary circulatory and total distribution volume. The difference between these volumes is assumed to be the volume of the SCS. From the dilution curve, it was also possible to calculate flow rates between and within the systems. The results of these experiments suggest a plasma volume in the PCS of 3.42+/-0.89 ml 100 g(-1) body mass, and in the SCS of 1.68+/-0.35 ml 100 g(-1) body mass (mean +/- S.D.) or approximately 50% that of the PCS. Flow rates to the SCS were calculated as 2.7% of the resting cardiac output. There was an allometric relationship between body mass and blood volumes. Increasing condition factor showed a tendency towards smaller blood volumes of the PCS, expressed as percentage body mass, but this was not evident for the volume of the SCS.

Retinal specializations in the blue marlin: eyes designed for sensitivity to low light levels

The large eyes and well-developed visual system of billfishes suggest that vision is an important sense for the detection and interception of prey and lures. Investigations of visual abilities in these large pelagic fishes are difficult, however anatomical studies of billfish eyes and retinas allow prediction of a number of visual capabilities. From the density of ganglion cells in the blue marlin (Makaira nigricans) retina, visual acuities of less than 10 cycles per degree were derived, a surprisingly low visual resolution given the absolute size of the marlin eye. Cone photoreceptors, on the other hand, were present in high densities, resulting in a presumed summation of cones to ganglion cells at a ratio of 40:1, even in the area of best vision. The optical sensitivity of the marlin eye was high owing to the large dimensions of the cone photoreceptors. These results indicate that the marlin eye is specifically adapted to cope with the low light levels encountered during diving. Since the marlin is likely to use its vision at depth, it is suggested that this line of research could help estimate the limits of vertical distribution based on light level.

Five generations of peace operations: from the "thin blue line" to "painting a country blue"

This article combines both international practice and analytical contributions into a systematic and synthetic presentation of the evolution of peace operations from their modern inception in 1948 to the present. It seeks to serve a didactic purpose in proposing a basic structure for Brazilian scholars' burgeoning debate on peace operations and intervention, rather than a definitive characterization of blue helmet practice. Peace operations' progression is traced through five analytical "generations," each adding a crucial factor distinguishing it from its predecessors. Each generation is placed in relation to changes in the nature of conflict and in the interpretation of the foundational principles of peace operations, and links to broader theoretical issues in International Relations are made explicit at each stage.

Blue-enhanced thin-film photodiode for dual-screen x-ray imaging

This article reports on a-Si:H-based low-leakage blue-enhanced photodiodes for dual-screen x-ray imaging detectors. Doped nanocrystalline silicon was incorporated in both the n- and p-type regions to reduce absorption losses for light incoming from the top and bottom screens. The photodiode exhibits a dark current density of 900 pA/cm(2) and an external quantum efficiency up to 90% at a reverse bias of 5 V. In the case of illumination through the tailored p-layer, the quantum efficiency of 60% at a 400 nm wavelength is almost double that for the conventional a-Si:H n-i-p photodiode.

Solubility of red 153 and blue 1 in supercritical carbon dioxide

Solubilities of red 153, (3-[[4-[[5,6(or 6,7)-dichloro-2-benzothiazolyl]azo]phenyl]ethylamino]propanenitrile), an azo compound, and disperse blue1 (1,4,5,8-tetraaminoantraquinone) in supercritical carbon dioxide (SC CO(2)) were measured at T = (333.2 to 393.2) K over the pressure range (12.0 to 40.0) MPa by a flow type apparatus. The solubility of red 153 (0.985. 10(-6) to 37.2. 10(-6)) in the overall region of measurements is found to be significantly higher than that of disperse blue 1 (1.12.10(-7) to 4.89.10(-7)). The solubility behavior of disperse red 153 follows the general solubility trend displayed by disperse dyes with a crossover pressure at about 20 MPa. On the other hand, blue 1, which is a disperse anthraquinone dye, exhibits unexpected behavior not recorded previously there is no crossover pressure at the temperature and pressure ranges studied, and the dye's solubility at T = 333.2 K practically does not increase with pressure. To the best of our knowledge, there are no previous measurements of blue 1 solubility in SC CO(2) reported in the literature. The experimental data were correlated by using the Soave Redlich Kwong equation of state (EoS) with the one-fluid van der Waals mixing rule, and an acceptable correlation of the solubility data for both dyes was obtained.

Blue rayon e teste Salmonella/microssoma na avaliação da qualidade de águas costeiras

OBJETIVO: Desenvolver estratégia para o monitoramento passivo das águas do estuário de Santos quanto à presença de atividade genotóxica e de hidrocarbonetos policíclicos aromáticos. MÉTODOS: Estudo realizado no estuário de Santos, Estado de São Paulo, em 2002. Foram selecionados e avaliados dois pontos de amostragem com diferentes graus de contaminação em duas campanhas de amostragem, utilizando a técnica de blue rayon in situ, análises químicas e o ensaio de Salmonella/microssoma com as linhagens bacterianas sensíveis a diferentes classes de compostos. Os extratos foram submetidos ao teste de Salmonella/microssoma em microssuspensão com as linhagens TA98, TA100, YG1041 e YG1042 na presença e ausência de ativação metabólica, e a análises químicas. RESULTADOS: O ponto 1, que apresentou sedimento com altas concentrações de hidrocarbonetos policíclicos aromáticos, mostrou maior freqüência de resultados positivos para o ensaio Samonella/microssoma e maiores concentrações de hidrocarbonetos policíclicos aromáticos em ambas as campanhas em comparação com o ponto 2, menos contaminado. A linhagem que se mostrou mais sensível foi a YG1041, que permitiu comparações entre locais com diferentes graus de contaminação. CONCLUSÕES: A combinação da técnica de blue rayon in situ com o ensaio Salmonella/microsoma com a linhagem YG1041 e as análises químicas se mostraram eficientes. Foi possível recuperar os compostos genotóxicos, e os hidrocarbonetos policíclicos aromáticos analisados, parecendo ser uma estratégia adequada para o monitoramento da qualidade das águas do estuário de Santos.

An Unprovoked Attack by a Blue Shark Prionace glauca (Chondrichthyes: Carcharhinidae) on a Spear Fisherman in Terceira Island, Azores, Northeast Atlantic

Copyright © 2014 Elsevier Science Ltd.

Role of the wider Azores region as a nursery ground for North Atlantic blue shark (Prionace glauca)

Tese de Doutoramento, Ciências do Mar (Ecologia Marinha)

Estudo da obtenção de gelatina a partir de raspa Wet-Blue da indústria de curtumes

Mestrado em Engenharia Química. Ramo Tecnologias de Protecção Ambiental.

Synthesis and properties of Co-doped titanate nanotubes and their optical sensitization with methylene blue

Agência Financiadora - Fundação para a Ciência e Tecnologia - PTDC/CTM NAN/113021/2009

Laccase–Prussian blue film–graphene doped carbon paste modified electrode for carbamate pesticides quantification

A novel enzymatic biosensor for carbamate pesticides detection was developed through the direct immobilization of Trametes versicolor laccase on graphene doped carbon paste electrode functionalized with Prussianblue films (LACC/PB/GPE). Graphene was prepared by graphite sonication-assisted exfoliation and characterized by transmission electron microscopy and X-ray photoelectron spectro- scopy. The Prussian blue film electrodeposited onto graphene doped carbon paste electrode allowed considerable reduction of the charge transfer resistance and of the capacitance of the device.The combined effects of pH, enzyme concentration and incubation time on biosensor response were optimized using a 23 full-factorial statistical design and response surface methodology. Based on the inhibition of laccase activity and using 4-aminophenol as redox mediator at pH 5.0,LACC/PB/GPE exhibited suitable characteristics in terms of sensitivity, intra-and inter-day repeatability (1.8–3.8% RSD), reproducibility (4.1 and 6.3%RSD),selectivity(13.2% bias at the higher interference: substrate ratios tested),accuracy and stability(ca. twenty days)for quantification of five carbamates widely applied on tomato and potato crops.The attained detection limits ranged between 5.2×10−9 mol L−1(0.002 mg kg−1 w/w for ziram)and 1.0×10−7 mol L−1 (0.022 mg kg−1 w/w for carbofuran).Recovery values for the two tested spiking levels ranged from 90.2±0.1%(carbofuran)to 101.1±0.3% (ziram) for tomato and from 91.0±0.1%(formetanate)to 100.8±0.1%(ziram)for potato samples.The proposed methodology is appropriate to enable testing pesticide levels in food samples to fit with regulations and food inspections.

Glucose-6-phosphate dehydrogenase and glutathione reductase activity in methemoglobin reduction by methylene blue and cyst amine: study on glucose-6-phosphate dehydrogenase-deficient individuals, on normal subjects and on riboflavin-treated subjects

The authors have standardized methods for evaluation of the activity of the glucose-6-phosphate dehydrogenase and of glutathione reductase. The general principle of the first method was based on methemoglobin formation by sodium nitrite followed by stimulation of the glucose-6-phosphate dehydrogenase with methylene blue. Forty six adults (23 males and 23 females) were studied. Subjects were not G6PD deficient and were aged 20 to 30 years. The results showed that methemoglobin reduction by methylene blue was 154.40 and 139.90 mg/min (p<0.05) for males and females, respectively, in whole blood, and 221.10 and 207.85 mg/min (n.s.), respectively, in washed red cells. These data showed that using washed red cells and 0.7g% sodium nitrite concentration produced no differences between sexes and also shortened reading time for the residual amount of methemoglobin to 90 minutes. Glutathione reductase activity was evaluated on the basis of the fact that cystamine (a thiol agent) binds to the SH groups of hemoglobin, forming complexes. These complexes are reversed by the action of glutathione reductase, with methemoglobin reduction occurring simultaneously with this reaction. Thirty two adults (16 males and 16 females) were studied. Subjects were not G6PD deficient and were aged 20 to 30 years. Methemoglobin reduction by cystamine was 81.27 and 91.13 mg/min (p<0.01) for males and females, respectively. These data showed that using washed red cells and 0.1 M cystamine concentration permits a reading of the residual amount of methemoglobin at 180 minutes of incubation. Glutathione reductase activity was evaluated by methemoglobin reduction by cystamine in 14 females before and after treatment with 10 mg riboflavin per day for 8 days. The results were 73.69 and 94.26 jug/min (p<0.01) before and after treatment, showing that riboflavin treatment increase glutathione reductase activity even in normal individuals. Three Black G6PD-deficient individuals (2 males and 1 female) were also studied. The G6PD and glutathione reductase were partially activated, the change being more intense in males. On the basis of race and of the laboratory characteristics observed, it is possible to suggest that the G6PD deficiency of these individuals is of the African type and that the female is heterozygous for this deficiency. Analysis of the results as a whole permitted us to conclude that the methods proposed here were efficient for evaluating the activity of the glucose-6-phosphate dehydrogenase and of glutathione reductase. The latter is dependent on the pentose pathway, which generates NADPH, and on riboflavin, a FAD precursor vitamin.

A novel colorimetric assay of beta-D-glucans in basidiomycete strains by alcian blue dye in a 96-well microtiter plate

Basidiomycete strains synthesize several types of beta-D-glucans, which play a major role in the medicinal properties of mushrooms. Therefore, the specific quantification of these beta-D-glucans in mushroom strains is of great biochemical importance. Because published assay methods for these beta-D-glucans present some disadvantages, a novel colorimetric assay method for beta-D-glucan with alcian blue dye was developed. The complex formation was detected by following the decrease in absorbance in the range of 620 nm and by hypsochromic shift from 620 to 606 nm (similar to 14 nm) in UV-Vis spectrophotometer. Analysis of variance was used for optimization of the slope of the calibration curve by using the assay mixture containing 0.017% (w/v) alcian blue in 2% (v/v) acetic acid at pH 3.0. The high-throughput colorimetric assay method on microtiter plates was used for quantification of beta-D-glucans in the range of 0-0.8 mu g, with a slope of 44.15 x 10(-2) and a limit of detection of 0.017 mu g/well. Recovery experiments were carried out by using a sample of Hericium erinaceus, which exhibited a recovery of 95.8% for beta-1,3-D-glucan. The present assay method exhibited a 10-fold higher sensitivity and a 59-fold lower limit of detection compared with the published method with congo red beta-D-glucans of several mushrooms strains were isolated from fruiting bodies and mycelia, and they were quantified by this assay method. This assay method is fast, specific, simple, and it can be used to quantify beta-D-glucans from other biological sources. (C) 2015 American Institute of Chemical Engineers

TEC4SEA - A Modular Platform for Research, Test and Validation of Technologies Supporting a Sustainable Blue Economy

This paper presents the TEC4SEA research infrastructure created in Portugal to support research, development, and validation of marine technologies. It is a multidisciplinary open platform, capable of supporting research, development, and test of marine robotics, telecommunications, and sensing technologies for monitoring and operating in the ocean environment. Due to the installed research facilities and its privileged geographic location, it allows fast access to deep sea, and can support multidisciplinary research, enabling full validation and evaluation of technological solutions designed for the ocean environment. It is a vertically integrated infrastructure, in the sense that it possesses a set of skills and resources which range from pure conceptual research to field deployment missions, with strong industrial and logistic capacities in the middle tier of prototype production. TEC4SEA is open to the entire scientific and enterprise community, with a free access policy for researchers affiliated with the research units that ensure its maintenance and sustainability. The paper describes the infrastructure in detail, and discusses associated research programs, providing a strategic vision for deep sea research initiatives, within the context of both the Portuguese National Ocean Strategy and European Strategy frameworks.