Structure of an Odorant-Binding Protein from the Mosquito Aedes aegypti Suggests a Binding Pocket Covered by a pH-Sensitive ""Lid""

Background: The yellow fever mosquito, Aedes aegypti, is the primary vector for the viruses that cause yellow fever, mostly in tropical regions of Africa and in parts of South America, and human dengue, which infects 100 million people yearly in the tropics and subtropics. A better understanding of the structural biology of olfactory proteins may pave the way for the development of environmentally-friendly mosquito attractants and repellents, which may ultimately contribute to reduction of mosquito biting and disease transmission. Methodology: Previously, we isolated and cloned a major, female-enriched odorant-binding protein (OBP) from the yellow fever mosquito, AaegOBP1, which was later inadvertently renamed AaegOBP39. We prepared recombinant samples of AaegOBP1 by using an expression system that allows proper formation of disulfide bridges and generates functional OBPs, which are indistinguishable from native OBPs. We crystallized AaegOBP1 and determined its three-dimensional structure at 1.85 angstrom resolution by molecular replacement based on the structure of the malaria mosquito OBP, AgamOBP1, the only mosquito OBP structure known to date. Conclusion: The structure of AaegOBP1 (= AaegOBP39) shares the common fold of insect OBPs with six alpha-helices knitted by three disulfide bonds. A long molecule of polyethylene glycol (PEG) was built into the electron-density maps identified in a long tunnel formed by a crystallographic dimer of AaegOBP1. Circular dichroism analysis indicated that delipidated AaegOBP1 undergoes a pH-dependent conformational change, which may lead to release of odorant at low pH (as in the environment in the vicinity of odorant receptors). A C-terminal loop covers the binding cavity and this ""lid"" may be opened by disruption of an array of acid-labile hydrogen bonds thus explaining reduced or no binding affinity at low pH.

Biotransformations of Substituted Phenylethanols and Acetophenones by Environmental Bacteria

Whole cells of hydrocarbon-degrading bacteria, isolated from polluted sediments in the Santos Estuary (Baixada Santista, Sao Paulo, Brazil), were able to catalyse oxidoreduction reactions with various substituted phenylethanols and acetophenones as substrates. A number of substituted phenylethanols were formed with high (>99 %) enantiomeric excess. The results of microbial oxidation of phenylethanols 2, 3, 5-7 by Acinetobacter sp. 6.4T and the reduction of acetophenones 1a-6a by Serratia marcescens 5.4T showed that the bacteria used as biocatalysts in this study present significant potential for exploitation in biotechnological processes. The reduction of prochiral acetophenones by Serratia marcescens 3.5T yielded optically active alcohols with 90-99 % enantiomeric excess, and Acinetobacter sp. 6.4T is a potential biocatalyst for the oxidation of alcohols.

Antimicrobial mechanisms behind photodynamic effect in the presence of hydrogen peroxide

This study describes the use of methylene blue (MB) plus light (photodynamic inactivation, PDI) in the presence of hydrogen peroxide (H(2)O(2)) to kill Staphylococcus aureus, Escherichia coli, and Candida albicans. When H(2)O(2) was added to MB plus light there was an increased antimicrobial effect, which could be due to a change in the type of ROS generated or increased microbial uptake of MB. To clarify the mechanism, the production of ROS was investigated in the presence and absence of H(2)O(2). It was observed that ROS production was almost inhibited by the presence of H(2)O(2) when cells were not present. In addition, experiments using different sequence combinations of MB and H(2)O(2) were performed and MB optical properties inside the cell were analyzed. Spectroscopy experiments suggested that the amount of MB was higher inside the cells when H(2)O(2) was used before or simultaneously with PDI, and ROS formation inside C. albicans cells confirmed that ROS production is higher in the presence of H(2)O(2). Moreover enzymatic reduction of MB by E. coli during photosensitizer uptake to the photochemically inactive leucoMB could be reversed by the oxidative effects of hydrogen peroxide, increasing ROS formation inside the microorganism. Therefore, the combination of a photosensitizer such as MB and H(2)O(2) is an interesting approach to improve PDI efficiency.

Quantification of some nonsteroidal anti-inflammatory drugs as reducing agents of Cu(II)/4,4 '-dicarboxy-2,2 '-biquinoline complexes in cationic micellar medium

A simple method was developed for spectrophotometric determination of some nonsteroidal anti-inflammatory drugs (meloxicam, piroxicam and tenoxicam) based on the reduction of copper(II) in buffered solution (pH 7.0) and micellar medium containing 4,4'-dicarboxy-2,2'-buffered solution (pH 7.0) and micellar medium containing 4,4'-dicarboxy-2,2'-biquinoline acid. The-biquinoline acid. The absorbance values at 558 nm, characteristic of the formed Cu(I)/4,4'-dicarboxy-2,2'-biquinoline complexes, are linear with the concentrations (5.7-40 mmol L(-1), n = 5) of these oxicams (meloxicam r = 0.998; piroxicam and tenoxicam r = 0.999). The limit of detection values, in mmol L(-1), calculated for meloxicam (2.7), piroxicam (1.2) and tenoxicam (1.3) was obtained with 99% confidence level and the relative standard deviations for meloxicam (3.1%), piroxicam (5.1%) and tenoxicam (1.2%) were calculated using a 25 mmol L(-1) solution (n = 7). Mean recovery values for meloxicam, piroxicam and tenoxicam forms were 100 +/- 6.9, 98.6 +/- 3.6 and 99.4 +/- 2.5%, respectively. The conditional potential of Cu(II)/Cu(I) in complex medium of 7.5 mmol L(-1) BCA was determined to be 629 +/- 11 mV vs. NHE.

Surface structure effects on the electrochemical oxidation of ethanol on platinum single crystal electrodes

Ethanol oxidation has been studied on Pt(111), Pt(100) and Pt(110) electrodes in order to investigate the effect of the surface structure and adsorbing anions using electrochemical and FTIR techniques. The results indicate that the surface structure and anion adsorption affect significantly the reactivity of the electrode. Thus, the main product of the oxidation of ethanol on the Pt(111) electrode is acetic acid, and acetaldehyde is formed as secondary product. Moreover, the amount of CO formed is very small, and probably associated with the defects present on the electrode surface. For that reason, the amount of CO(2) is also small. This electrode has the highest catalytic activity for the formation of acetic acid in perchloric acid. However, the formation of acetic acid is inhibited by the presence of specifically adsorbed anions, such as (bi) sulfate or acetate, which is the result of the formation of acetic acid. On the other hand, CO is readily formed at low potentials on the Pt(100) electrode, blocking completely the surface. Between 0.65 and 0.80 V, the CO layer is oxidized and the production of acetaldehyde and acetic acid is detected. The Pt(110) electrode displays the highest catalytic activity for the splitting of the C-C bond. Reactions giving rise to CO formation, from either ethanol or acetaldehyde, occur at high rate at any potential. On the other hand, the oxidation of acetaldehyde to acetic acid has probably the lower reaction rate of the three basal planes.

Green Chemistry-Sensitive Analytical Procedure for Photometric Determination of Orthophosphate in River and Tap Water by Use of a Simple LED-Based Photometer

In the current work a Green Analytical Chemistry (GAC) procedure for photometric determination of orthophosphate in river water at mu g L-1 concentration level is described. The flow system module and the LED-based photometer were assembled together to constitute a compact unit in order to allow that a flow cell with optical path-length of 100mm was coupled to them. The photometric procedure based on the molybdenum blue method was implemented employing the multicommuted flow injection analysis approach, which provided facilities to allow reduction of reagent consumption and as well as waste generation. Aiming to prove the usefulness of the system, orthophosphate in river and tap waters was determined. Accuracy was ascertained by spiking samples with orthophosphate solution yielding recoveries ranging from 96% up to 107%. Other profitable features such as a wide linear response range between 10 to 800 mu g L-1 [image omitted]; a detection limit (3 sigma criterion) of 2.4 mu g L-1 [image omitted]; a relative standard deviation (n=7) of 2% using a typical water sample with concentration of 120 mu g L-1 [image omitted]; reagent consumption of 3.0mg ammonium molybdate, 0.3mg hydrazine sulfate, and 0.03mg stannous chloride per determination; a waste generation of 2.4mL per determination; and a sampling throughput of 20 determination per hours were also achieved.

Fermentation of cacao (Theobroma cacao L.) seeds with a hybrid Kluyveromyces marxianus strain improved product quality attributes

Fermentation of Theobroma cacao (cacao) seeds is an absolute requirement for the full development of chocolate flavor precursors. An adequate aeration of the fermenting cacao seed mass is a fundamental prerequisite for a satisfactory fermentation. Here, we evaluated whether a controlled inoculation of cacao seed fermentation using a Kluyveromyces marxianus hybrid yeast strain, with an increased pectinolytic activity, would improve an earlier liquid drainage (`sweatings`) from the fermentation mass, developing a superior final product quality. Inoculation with K. marxianus increased by one third the volume of drained liquid and affected the microorganism population structure during fermentation, which was detectable up to the end of the process. Introduction of the hybrid yeast affected the profile of total seed protein degradation evaluated by polyacrylamide gel electrophoresis, with improved seed protein degradation, and reduction of titrable acidity. Sensorial evaluation of the chocolate obtained from beans fermented with the K. marxianus inoculation was more accepted by analysts in comparison with the one from cocoa obtained through natural fermentation. The increase in mass aeration during the first 24 h seemed to be fundamental for the improvement of fermentation quality, demonstrating the potential application of this improved hybrid yeast strain with superior exogenous pectinolytic activity.

Low nanomolar concentration of mercury chloride increases vascular reactivity to phenylephrine and local angiotensin production in rats

Exposure to mercury at nanomolar level affects cardiac function but its effects on vascular reactivity have yet to be investigated. Pressor responses to phenylephrine (PHE) were investigated in perfused rat tail arteries before and after treatment with 6 nM HgCl2 during 1 h,,in the presence (E+) and absence (E-) of endothelium, after L-NAME (10(-4) M), indomethacin (10(-5) M), enalaprilate (1 mu M), tempol (1 mu M) and deferoxamine (300 mu M) treatments. HgCl2 increased sensitivity (pD(2)) without modifying the maximum response (Em) to PHE, but the pD(2) increase was abolished after endothelial damage. L-NAME treatment increased pD(2) and Emax. However, in the presence of HgCl2, this increase was smaller, and it did not modify Emax. After indomethacin treatment, the increase of pD(2) induced by HgCl2 was maintained. Enalaprilate, tempol and deferoxamine reversed the increase of pD(2) evoked by HgCl2. HgCl2 increased the angiotensin converting enzyme (ACE) activity explaining the result obtained with enalaprilate. Results suggest that at nanomolar concentrations HgCl2 increase the vascular reactivity to PHE. This response is endothelium mediated and involves the reduction of NO bioavailability and the action of reactive oxygen species. The local ACE participates in mercury actions and depends on the angiotensin 11 generation. (c) 2007 Elsevier Inc. All rights reserved.

Biological detoxification of different hemicellulosic hydrolysates using Issatchenkia occidentalis CCTCC M 206097 yeast

This work had as its main objective to contribute to the development of a biological detoxification of hemicellulose hydrolysates obtained from different biomass plants using Issatchenkia occidentalis CCTCC M 206097 yeast. Tests with hemicellulosic hydrolysate of sugarcane bagasse in different concentrations were carried out to evaluate the influence of the hydrolysate concentration on the inhibitory compounds removal from the sugarcane bagasse hydrolysate, without reduction of sugar concentration. The highest reduction values of inhibitors concentration and less sugar losses were observed when the fivefold concentrated hydrolysate was treated by the evaluated yeast. In these experiments it was found that the high sugar concentrations favored lower sugar consumption by the yeast. The highest concentration reduction of syringaldehyde (66.67%), ferulic acid (73.33%), furfural (62%), and 5-HMF (85%) was observed when the concentrated hydrolysate was detoxified by using this yeast strain after 24 h of experimentation. The results obtained in this work showed the potential of the yeast Issatchenkia occidentalis CCTCC M 206097 as detoxification agent of hemicellulosic hydrolysate of different biomass plants.

Modeling and analysis of laminate composite plates with embedded active-passive piezoelectric networks

The objective of this work is to present the finite element modeling of laminate composite plates with embedded piezoelectric patches or layers that are then connected to active-passive resonant shunt circuits, composed of resistance, inductance and voltage source. Applications to passive vibration control and active control authority enhancement are also presented and discussed. The finite element model is based on an equivalent single layer theory combined with a third-order shear deformation theory. A stress-voltage electromechanical model is considered for the piezoelectric materials fully coupled to the electrical circuits. To this end, the electrical circuit equations are also included in the variational formulation. Hence, conservation of charge and full electromechanical coupling are guaranteed. The formulation results in a coupled finite element model with mechanical (displacements) and electrical (charges at electrodes) degrees of freedom. For a Graphite-Epoxy (Carbon-Fibre Reinforced) laminate composite plate, a parametric analysis is performed to evaluate optimal locations along the plate plane (xy) and thickness (z) that maximize the effective modal electromechanical coupling coefficient. Then, the passive vibration control performance is evaluated for a network of optimally located shunted piezoelectric patches embedded in the plate, through the design of resistance and inductance values of each circuit, to reduce the vibration amplitude of the first four vibration modes. A vibration amplitude reduction of at least 10 dB for all vibration modes was observed. Then, an analysis of the control authority enhancement due to the resonant shunt circuit, when the piezoelectric patches are used as actuators, is performed. It is shown that the control authority can indeed be improved near a selected resonance even with multiple pairs of piezoelectric patches and active-passive circuits acting simultaneously. (C) 2010 Elsevier Ltd. All rights reserved.

Theoretical and experimental study of the thermal degradation of Eucalyptus timber

Thermal action on timber causes it to degrade through combustion of its chemical components, which leads to the release of vapors, combustible gases and surface char. This diminishes its load capacity, due to the reduction of its cross section by charring and to changes in its mechanical properties of strength and stiffness as a function of its exposure to high temperatures. This paper reports the charring rates observed on Eucalyptus structural beams and presents a numerical and experimental study of the behavior of these beams when exposed to fire, in which the properties of strength and stiffness were evaluated as a function of rising temperatures, allowing an analysis of the effect of the section factor on the internal rise in temperature of structural Eucalyptus beams.

Influence of feed time and sulfate load on the organic and sulfate removal in an ASBR

The removal of sulfate and organic matter was assessed in an ASBR, which treated wastewater containing 500 mg COD L(-1) (3 g COD L(-1) d(-1)) in 8 h-cycles at 30 degrees C. The wastewater was enriched with sulfate at [COD/SO(4)(2-]) ratios of 1.34, 0.67 and 0.34 (8.8,4.5 and 2.2 gSO(4)(2-) L(-1) d(-1)). For each COD/[SO(4)(2-)] ratio fill times used were: 10 min (batch), 3 and 6 h (fed-batch), achieving sulfate reduction of 30%, 72% and 72% (COD/[SO(4)(2-)] of 1.34); 25%, 58% and 55% (COD/[SO(4)(2-)] of 0.67) and 23%, 37% and 27% (COD/[SO(4)(2-)] of 0.34), respectively, and organic matter removal of 87%, 68% and 80% (COD/[SO(4)(2-)] of 1.34); 78%, 75% and 69% (COD/[SO(4)(2-)] of 0.67) and 85%, 84% and 83% (COD/[SO(4)(2-)] of 0.34), respectively. The results showed that fed-batch operation improved sulfate reduction, whereas organic matter removals were similar for batch and fed-batch operation. In addition, increase in sulfate loading in the fed-batch operation improved organic matter removal. (C) 2010 Elsevier Ltd. All rights reserved.

Treatment of linear alkylbenzene sulfonate in a horizontal anaerobic immobilized biomass reactor

Linear alkylbenzene sulfonate (LAS) is an anionic surfactant widely used to manufacture detergents and found in domestic and industrial wastewater. LAS removal was evaluated in a horizontal anaerobic immobilized biomass reactor. The system was filled with polyurethane foam and inoculated with sludge that was withdrawn from an up flow anaerobic sludge blanket reactor that is used to treat swine wastewater. The reactor was fed with easily degradable substrates and a solution of commercial LAS for 313 days. The hydraulic retention time applied was 12 h. The system was initially operated without detergent and resulted to 94% reduction of demand. The mass balance in the system indicated that the LAS removal efficiency was 45% after 180 days. From the 109th day to the 254th day, a removal efficiency of 32% was observed. The removal of LAS was approximately 40% when 1500 mg of LAS were applied in the absence of co-substrates suggesting that the LAS molecules were used selectively. Microscopic analyses of the biofilm revealed diverse microbial morphologies and denaturing gradient gel electrophoresis profiling showed variations in the total bacteria and sulfate-reducing bacteria populations. 16S rRNA sequencing and phylogenetic analyses demonstrated that members of the order Clostridiales were the major components of the bacterial community in the last step of the reactor operation. (c) 2009 Elsevier Ltd. All rights reserved.

Greenhouse gas emissions and energy balance of palm oil biofuel

The search for alternatives to fossil fuels is boosting interest in biodiesel production. Among the crops used to produce biodiesel, palm trees stand out due to their high productivity and positive energy balance. This work assesses life cycle emissions and the energy balance of biodiesel production from palm oil in Brazil. The results are compared through a meta-analysis to previous published studies: Wood and Corley (1991) [Wood BJ, Corley RH. The energy balance of oil palm cultivation. In: PORIM intl. palm oil conference agriculture; 1991.], Malaysia; Yusoff and Hansen (2005) [Yusoff S. Hansen SB. Feasibility study of performing an life cycle assessment on crude palm oil production in Malaysia. International Journal of Life Cycle Assessment 2007;12:50-8], Malaysia; Angarita et al. (2009) [Angarita EE, Lora EE, Costa RE, Torres EA. The energy balance in the palm oil-derived methyl ester (PME) life cycle for the cases in Brazil and Colombia. Renewable Energy 2009;34:2905-13], Colombia; Pleanjai and Gheewala (2009) [Pleanjai S. Gheewala SH. Full chain energy analysis of biodiesel production from palm oil in Thailand. Applied Energy 2009;86:S209-14], Thailand; and Yee et al. (2009) [Yee KF, Tan KT, Abdullah AZ, Lee la. Life cycle assessment of palm biodiesel: revealing facts and benefits for sustainability. Applied Energy 2009;86:S189-96], Malaysia. In our study, data for the agricultural phase, transport, and energy content of the products and co-products were obtained from previous assessments done in Brazil. The energy intensities and greenhouse gas emission factors were obtained from the Simapro 7.1.8. software and other authors. These factors were applied to the inputs and outputs listed in the selected studies to render them comparable. The energy balance for our study was 1:5.37. In comparison the range for the other studies is between 1:3.40 and 1:7.78. Life cycle emissions determined in our assessment resulted in 1437 kg CO(2)e/ha, while our analysis based on the information provided by other authors resulted in 2406 kg CO(2)e/ha, on average. The Angarita et al. (2009) [Angarita EE, Lora EE, Costa RE, Torres EA. The energy balance in the palm oil-derived methyl ester (PME) life cycle for the cases in Brazil and Colombia. Renewable Energy 2009:34:2905-13] study does not report emissions. When compared to diesel on a energy basis, avoided emissions due to the use of biodiesel account for 80 g CO(2)e/MJ. Thus, avoided life Cycle emissions associated with the use of biodiesel yield a net reduction of greenhouse gas emissions. We also assessed the carbon balance between a palm tree plantation, including displaced emissions from diesel, and a natural ecosystem. Considering the carbon balance outcome plus life cycle emissions the payback time for a tropical forest is 39 years. The result published by Gibbs et al. (2008) [Gibbs HK, Johnston M, Foley JA, Holloway T, Monfreda C, Ramankutty N, et al., Carbon payback times for crop-based biofuel expansion in the tropics: the effects of changing yield and technology. Environmental Research Letters 2008;3:10], which ignores life cycle emissions, determined a payback range for biodiesel production between 30 and 120 years. Crown Copyright (C) 2010 Published by Elsevier Ltd. All rights reserved.

Analysis of the emissions of volatile organic compounds from the compression ignition engine fueled by diesel-biodiesel blend and diesel oil using gas chromatography

This paper describes the procedures of the analysis Of Pollutant gases, as volatile organic compounds (benzene, toluene, ethylbenzene, o-xylene, m-xylene and p-xylene) emitted by engines, using high-resolution gas chromatography (HRGC). In a broad sense, CI engine burning diesel was compared with B10 and a drastic reduction was observed in the emissions of the aromatic compounds by using B10. Especially for benzene, the reduction of concentrations occurs on the level of about 19.5%. Although a concentration value below 1 mu g ml(-1) has been obtained, this reduction is extremely significant since benzene is a carcinogenic compound. (c) 2008 Elsevier Ltd. All rights reserved.