Crystallization and preliminary X-ray diffraction analysis of recombinant chlorocatechol 1,2-dioxygenase from Pseudomonas putida

Chlorocatechol 1,2-dioxygenase from the Gram-negative bacterium Pseudomonas putida (Pp 1,2-CCD) is considered to be an important biotechnological tool owing to its ability to process a broad spectrum of organic pollutants. In the current work, the crystallization, crystallographic characterization and phasing of the recombinant Pp 1,2-CCD enzyme are described. Reddish-brown crystals were obtained in the presence of polyethylene glycol and magnesium acetate by utilizing the vapour-diffusion technique in sitting drops. Crystal dehydration was the key step in obtaining data sets, which were collected on the D03B-MX2 beamline at the CNPEM/MCT - LNLS using a MAR CCD detector. Pp 1,2-CCD crystals belonged to space group P6(1)22 and the crystallographic structure of Pp 1,2-CCD has been solved by the MR-SAD technique using Fe atoms as scattering centres and the coordinates of 3-chlorocatechol 1,2-dioxygenase from Rhodococcus opacus (PDB entry 2boy) as the search model. The initial model, which contains three molecules in the asymmetric unit, has been refined to 3.4 A resolution.

Amazon deforestation alters small stream structure, nitrogen biogeochemistry and connectivity to larger rivers

Human activities that modify land cover can alter the structure and biogeochemistry of small streams but these effects are poorly known over large regions of the humid tropics where rates of forest clearing are high. We examined how conversion of Amazon lowland tropical forest to cattle pasture influenced the physical and chemical structure, organic matter stocks and N cycling of small streams. We combined a regional ground survey of small streams with an intensive study of nutrient cycling using (15)N additions in three representative streams: a second-order forest stream, a second-order pasture stream and a third-order pasture stream. These three streams were within several km of each other and on similar soils. Replacement of forest with pasture decreased stream habitat complexity by changing streams from run and pool channels with forest leaf detritus (50% cover) to grass-filled (63% cover) channel with runs of slow-moving water. In the survey, pasture streams consistently had lower concentrations of dissolved oxygen and nitrate (NO(3) (-)) compared with similar-sized forest streams. Stable isotope additions revealed that second-order pasture stream had a shorter NH(4) (+) uptake length, higher uptake rates into organic matter components and a shorter (15)NH(4) (+) residence time than the second-order forest stream or the third-order pasture stream. Nitrification was significant in the forest stream (19% of the added (15)NH(4) (+)) but not in the second-order pasture (0%) or third-order (6%) pasture stream. The forest stream retained 7% of added (15)N in organic matter compartments and exported 53% ((15)NH(4) (+) = 34%; (15)NO(3) (-) = 19%). In contrast, the second-order pasture stream retained 75% of added (15)N, predominantly in grasses (69%) and exported only 4% as (15)NH(4) (+). The fate of tracer (15)N in the third-order pasture stream more closely resembled that in the forest stream, with 5% of added N retained and 26% exported ((15)NH(4) (+) = 9%; (15)NO(3) (-) = 6%). These findings indicate that the widespread infilling by grass in small streams in areas deforested for pasture greatly increases the retention of inorganic N in the first- and second-order streams, which make up roughly three-fourths of total stream channel length in Amazon basin watersheds. The importance of this phenomenon and its effect on N transport to larger rivers across the larger areas of the Amazon Basin will depend on better evaluation of both the extent and the scale at which stream infilling by grass occurs, but our analysis suggests the phenomenon is widespread.

Arsenic and Chromium in Brazilian Agricultural Supplies

Arsenic (As) and chromium (Cr) contents were measured in agricultural supplies used at different farms in Sao Paulo State, Brazil. The highest mass fractions of As were found in thermophosphates, reaching levels of 4 mg/kg. The highest mass fractions of Cr (21 g/kg) were found in calcium magnesium silicate, while the thermophosphates also presented high values reaching approximately 1 g/kg. The levels of As were within Brazilian guidelines, but the values of Cr in thermophosphates exceeded the levels permitted in Brazil. The As content in fertilizers may be considered safe (5 mg/kg) in terms of environmental pollution. However, the Cr content in calcium magnesium silicate following continuous use may constitute a significant problem in Brazil due to potentially increasing levels of this metal in soils.

Evaluation of laser induced breakdown spectrometry for the determination of macro and micronutrients in pharmaceutical tablets

The aim of this work is to demonstrate the feasibility of laser induced breakdown spectrometry (LIBS) for the determination of macro and micronutrients in multielement tablets. The experimental setup was designed by using a laser Q-switch (Nd:YAG, 10 Hz, lambda = 1064 nm) and the emission signals were collected by lenses into an optical fiber coupled to an echelle spectrometer equipped with a high-resolution intensified charge coupled device (ICCD). Tablets were cryogenically ground and thereafter pelletized before LIBS analysis. Calibration curves were made by employing samples and mixtures of commercial multielement tablets with binders at different ratios. Best results were achieved by using the following experimental conditions: 29 J cm(-2) laser fluence, 165 mm lens to sample distance (f = 200 mm), 2.0 mu s delay time, 5.0 mu s integration time and 5 accumulated laser pulses. In general, the results obtained by the proposed LIBS procedure were in agreement with those obtained by ICP OES from the corresponding acid digests and coefficients variation of LIBS measurements varied from 2 to 16%. The metrological figures of merit indicate that LIBS fits for the intended purposes, and can be recommended for the analysis of multielement tablets and similar matrices aiming the determination of Ca, Cu, Fe, Mg, Mn, P and Zn.

Sequential injections as an alternative to gradient exploitation for implementing differential kinetic analysis in a flow injection system

A novel flow-based strategy for implementing simultaneous determinations of different chemical species reacting with the same reagent(s) at different rates is proposed and applied to the spectrophotometric catalytic determination of iron and vanadium in Fe-V alloys. The method relies on the influence of Fe(II) and V(IV) on the rate of the iodide oxidation by Cr(VI) under acidic conditions, the Jones reducing agent is then needed Three different plugs of the sample are sequentially inserted into an acidic KI reagent carrier stream, and a confluent Cr(VI) solution is added downstream Overlap between the inserted plugs leads to a complex sample zone with several regions of maximal and minimal absorbance values. Measurements performed on these regions reveal the different degrees of reaction development and tend to be more precise Data are treated by multivariate calibration involving the PLS algorithm The proposed system is very simple and rugged Two latent variables carried out ca 95% of the analytical information and the results are in agreement with ICP-OES. (C) 2010 Elsevier B V. All rights reserved.

Adjustment of Mineral Elements in the Culture Medium for the Micropropagation of Three Vriesea Bromeliads from the Brazilian Atlantic Forest: The Importance of Calcium

Many different species of Bromeliaceae are endangered and their conservation requires specific knowledge of their growth habits and propagation. In vitro culture of bromeliads is an important method for efficient clonal propagation and ill vitro seed g,germination can be used to maintain genetic variability. The present work aims to evaluate the in vitro growth and nutrient concentration in leaves of the epiphyte bromeliads Vriesea friburguensis Mez, Vriesea hieroglyphica (Carriere) E. Morren, and Vriesea unilateralis Mez, which exhibit slow rates of growth in vivo and in vitro. Initially, we compared the endogenous mineral composition of bromeliad plantlets grown in half-strength Murashige and Skoog (MS) medium and the mineral composition considered adequate in the literature. This approach suggested that calcium (Ca) is a critical nutrient and this was considered for new media formulation. Three new culture media were defined in which the main changes to half-strength MS medium were an increase in Ca, magnesium, sulfur, copper, and chloride and a decrease in iron, maintaining the nitrate: ammonium rate at approximate to 2:1. The main difference among the three new media formulated was Ca concentration, which varied from 1.5 mm in half-strength MS to 3.0, 6.0, and 12 mm in M2, M3, and M4 media, respectively. Consistently, all three species exhibited significantly higher fresh and dry weight on M4, the newly defined medium with the highest level of Ca (12 mm). Leaf nitrogen, potassium, zinc, magnesium and boron concentrations increased as Ca concentration in the medium increased from 1.5 to 12 mm.

Macronutrients in Marandu Palisade Grass as Influenced by Lime, Slag, and Nitrogen Fertilization

Soil acidity is one of the main limiting factors for the growth of pasture grasses in Brazilian soils. In addition to lime, slag can be used to correct soil acidity and help plants to absorb nutrients in adequate amounts. The objective of this experiment was to evaluate, under greenhouse conditions, the effects of slag and lime plus nitrogen (N) on marandu palisade grass plants` nutritional status as well as the absorption of macronutrients submitted to two cuts. The treatments consisted of two corrective materials (slag and lime), three corrective material rates (0.81, 1.61, and 3.22 g dm-3 of ECaCO3), three N rates (75, 150, and 300 mg dm-3) plus a control treatment, with four replications. Macronutrient contents in the forage plants were found to be present in adequate levels. The mean value of N accumulated in the shoot was 40.1 mg per plant, phosphorus (P) was 4.6 mg per plant, potassium (K) was 38.6 mg per plant, calcium (Ca) was 7.3 mg per plant, magnesium (Mg) was 6.7 mg per plant, and sulfur (S) was 3.5 mg per plant at the first cut. At the second cut, the nutrient accumulations values were N 50.8 mg per plant, P 6.3 mg per plant, K 20.7 mg per plant, Ca 21.6 mg per plant, Mg 24.0 mg per plant, and S 4.7 mg per plant. Macronutrients accumulation in the shoot of grass increased with the addition of both the correctives as well as the N rates.

Production, microstructural characterization and mechanical properties of as-cast Ti-10Mo-xNb alloys

Beta titanium (Ti) alloys are one of the most promising groups of Ti alloys for biomedical applications. This work presents the production, microstructural characterization, and mechanical properties of as-cast Ti-10Mo-xNb (x = 0, 3, 6, 9, 20, and 30) alloys. They were produced via arc melting and characterized by scanning electron microscopy and X-ray diffraction. The density of each alloy was measured by the Archimedes method and the mechanical characterization was carried out by using the Vickers microhardness test and Young`s modulus measurements. The results show a near beta microstructure for niobium (Nb) contents lower or equal to 9 wt% while beta single-phase microstructure was obtained for higher Nb additions. The microhardness values decreased with the increase of Nb content in the alloys. The elastic modulus values of Ti-10Mo-3Nb and Ti-10Mo-20Nb alloys were lower than those of cp Ti and Ti-6Al-4V.

Utilization of pineapple stem juice to enhance enzyme-hydrolytic efficiency for sugarcane bagasse after an optimized pre-treatment with alkaline peroxide

The enzymatic hydrolysis of sugarcane bagasse was investigated by treating a peroxide-alkaline bagasse with a pineapple stem juice, xylanase and cellulase. Pre-treatment procedures of sugarcane bagasse with alkaline hydrogen peroxide were evaluated and compared. Analyses were performed using 2(4) factorial designs, with pre-treatment time, temperature, magnesium sulfate and hydrogen peroxide concentration as factors. The responses evaluated were the yield of cellobiose and glucose released from pretreated bagasse after enzymatic hydrolysis. The results show that the highest enzymatic conversion was obtained for bagasse using 2% hydrogen peroxide at 60 degrees C for 16 h in the presence of 0.5% magnesium sulfate. Bagasse (5%) was treated with pineapple stem extract, which contains mixtures of protease and esterase, in combination with xylanase and cellulase. It was observed that the amount of glucose and cellobiose released from bagasse increased with the mixture of enzymes. It is believed that the enzymes present in pineapple extracts are capable of hydrolyze specific linkages that would facilitate the action of digesting plant cell walls enzymes. This increases the amount of glucose and other hexoses that are released during the enzymatic treatment and also reduces the amount of cellulase necessary in a typical hydrolysis. (C) 2010 Elsevier Ltd. All rights reserved.

Search for optimum conditions of sugarcane bagasse hemicellulose extraction

A process has been elaborated for one-step low lignin content sugarcane bagasse hemicellulose extraction using alkaline solution of hydrogen peroxide. To maximize the hemicellulose yields several extraction conditions were examined applying the 2(4) factorial design: H(2)O(2) concentration from 2 to 6% (w/v), reaction time from 4 to 16 h, temperature from 20 to 60 degrees C, and magnesium sulfate absence or presence (0.5%, w/v). This approach allowed selection of conditions for the extraction of low and high lignin content hemicellulose. At midpoint the yield of hemicellulose was 94.5% with more than 88% of lignin removed. Lignin removal is suppressed at low extraction temperatures and in the absence of magnesium sulfate. Hemicellulose in 86% yield with low lignin content (5.9%) was obtained with 6% H(2)O(2) treatment for 4 h and 20 degrees C. This hemicellulose is much lighter in color than samples obtained at the midpoint condition and was found suitable for subsequent enzymatic hydrolysis. (C) 2009 Elsevier B.V. All rights reserved.

Thermal expansion of the V(5)Si(3) and T(2) phases of the V-Si-B system investigated by high-temperature X-ray diffraction

The thermal expansion anisotropy of the V(5)Si(3) and T(2)-phase of the V-Si-B system were determined by high-temperature X-ray diffraction from 298 to 1273 K. Alloys with nominal compositions V(62.5)Si(37.5) (V5Si3 phase) and V(63)Si(12)B(25) (T(2)-phase) were prepared from high-purity materials through arc-melting followed by heat-treatment at 1873 K by 24 h, under argon atmosphere. The V(5)Si(3) phase exhibits thermal expansion anisotropy equals to 1.3, with thermal expansion coefficients along the a and c-axis equal to 9.3 x 10(-6) K(-1) and 11.7 x 10(-6) K(-1), respectively. Similarly, the thermal expansion anisotropy value of the T(2)-phase is 0.9 with thermal expansion coefficients equal to 8.8 x 10(-6) K(-1) and 8.3 x 10(-6) K(-1) along the, a and c-axis respectively. Compared to other isostructural silicides of the 5:3 type and the Ti(5)Si(3) phase, the V(5)Si(3) phase presents lower thermal expansion anisotropy. The T(2)-phase present in the V-Si-B system exhibits low thermal expansion anisotropy, as the T(2)-phase of the Mo-Si-B, Nb-Si-B and W-Si-B systems. (C) 2009 Elsevier Ltd. All rights reserved.

Spontaneous infiltrations of compound systems of Y(2)O(3), Sm(2)O(3), RE(2)O(3), Al(2)O(3) and AIN in SiC ceramics

Silicon carbide ceramics (SiC) are used in different applications in the engineering area due to the excellent properties, mainly in high temperatures. They are usually obtained by liquid-phase sintering enabling to form volatile products and, consequently, defects. The present work aims at studying the obtention of SiC ceramics by spontaneous infiltration using a eutectic composition of the Al(2)O(3)/Y(2)O(3), AIN/Y(2)O(3), Al(2)O(3)/Sm(2)O(3), AlN/Sm(2)O(3), Al(2)O(3)/RE(2)O(3) and AlN/RE(2)O(3) Systems. RE(2)O(3) is the concentrate of the rare-earth oxide obtained from Xenotime ore. Infiltration tests were carried out in argon atmosphere, graphite crucibles, in several temperatures near the melting point of each system, varying from 2.5 to 60 min. It was observed that Al(2)O(3)/Y(2)O(3), Al(2)O(3)/SM(2)O(3), AlN/SM(2)O(3) and Al(2)O(3)/RE(2)O(3) systems do not infiltrate appropriately and the AlN/Y(2)O(3) and AlN/RE(2)O(3) systems infiltrated spontaneously more than 20 mm; however, the first one presented a higher degree of infiltration, approximately 97%. (C) 2009 Elsevier Ltd and Techna Group S.r.l. All rights reserved.

Glass formation of alloys selected by lambda and electronegativity criteria in the Ti-Zr-Fe-Co system

This work evaluates the glass formation of selected alloys based on the Ti-Zr-Fe-Co system, assuming the synergy of two distinct criteria: minimum topological instability and average electronegativity plots. Combining the minimum topological instability and the average electronegativity values result in a plot in which the most probable good glass former compositions are identified Ti-Zr rich alloys with Fe and Co additions were produced, compared against the final plot, and the best glass forming alloy composition was found to be very close the theoretically predicted ones on the Ti-Zr rich side, for both Ti-Zr-Fe and Ti-Zr-Co systems. (C) 2009 Elsevier B V All rights reserved

Citric acid as anti-hydration additive for magnesia containing refractory castables

Magnesia hydration is a key concern in refractory castable processing. The volumetric expansion that follows this reaction can result in cracks or even explosion during the first heating-up. Citric acid (CA) and other chelants can significantly reduce MgO hydration rate in aqueous suspensions by forming an insoluble magnesium citrate protective coating on the magnesia particles` surface. In the present work, the performance of CA as an anti-hydration additive in refractory castables was evaluated by hydration tests, mechanical strength and apparent volumetric expansion (AVE) measurements and thermogravimetry. The results attained have shown that CA effectiveness depends strongly on the amount added and by the interaction with other raw materials in the composition, in particular calcium aluminate cement. (C) 2011 Elsevier Ltd and Techna Group S.r.l. All rights reserved.

Porous alumina-spinel ceramics for high temperature applications

In order to reduce energy costs, high-temperature insulation porous refractory ceramics have been subjected to increasing demands. Among the techniques used to produce these materials (such as the addition of foaming agents and organic compounds), the pore generation via phase transformation presents key aspects, such as easy processing and the absence of toxic volatiles. In this study, this technique was applied to produce porous ceramics by decomposing an aluminum magnesium hydro-carbonate known as hydrotalcite (Mg(6)Al(2)(CO(3))(OH)(16)center dot 4H(2)O). It was found out that by using this complex compound, a large fraction of pores can be generated and kept at high temperatures (above 1300 degrees C) due to the in situ formation of spinel-like phases (MgAl(2)O(4)). (C) 2011 Elsevier Ltd and Techna Group S.r.l. All rights reserved.