Acute selenium selenite exposure effects on oxidative stress biomarkers and essential metals and trace-elements in the model organism zebrafish (Danio rerio).

Selenium (Se) is an essential trace-element that becomes toxic when present at high concentrations. Little is known regarding Se effects on parameters such as oxidative stress biomarkers. The aim of the present study was to investigate the effects of acute selenium exposure on oxidative stress biomarkers in a model organism, zebrafish (Danio rerio). Fish were exposed to selenium selenite at 1mgL(-1). Reduced glutathione (GSH), and metallothionein (MT) concentrations were determined in liver, kidney and brain, with MT also being determined in bile. Essential metals and trace-elements were also determined by inductively coupled mass spectrometry (ICP-MS) in order to verify possible metal homeostasis alterations. GSH concentrations in liver, kidney and brain increased significantly (1.05±0.03µmolg(-1) ww, 1.42±0.03µmolg(-1) ww and 1.64±0.03µmolg(-1) ww, respectively) in the Se-exposed group when compared to the controls (0.88±0.05µmolg(-1) ww, 0.80±0.04µmolg(-1) ww and 0.89±0.03µmolg(-1) ww for liver, kidney and brain, respectively). MT levels in Se-exposed liver (0.52±0.03µmolg(-1) ww) decreased significantly in comparison to the control group (0.64±0.02µmolg(-1) ww), while levels in bile increased, albeit non-significantly. This is in accordance with previous studies that indicate efficient biliary MT action, leading to a rapid metabolism and elimination of contaminants from the body. Levels in the brain increased significantly after Se-exposure (0.57±0.01µmolg(-1) ww) when compared to the control group (0.35±0.03µmolg(-1) ww) since this organ does not present a detoxification route as quick as the liver-gallbladder route. Several metal and trace-elements were altered with Se-exposure, indicating that excess of selenium results in metal dyshomeostasis. This is the first report on metal dyshomeostasis due to Se-exposure, which may be the first step in the mechanism of action of selenium toxicity, as is postulated to occur in certain major human pathophysiologies.

Electrophoresis and spectrometric analyses of adaptation-related proteins in thermally stressed Chromobacterium violaceum.

Chromobacterium violaceum is a Gram-negative proteobacteria found in water and soil; it is widely distributed in tropical and subtropical regions, such as the Amazon rainforest. We examined protein expression changes that occur in C. violaceum at different growth temperatures using electrophoresis and mass spectrometry. The total number of spots detected was 1985; the number ranged from 99 to 380 in each assay. The proteins that were identified spectrometrically were categorized as chaperones, proteins expressed exclusively under heat stress, enzymes involved in the respiratory and fermentation cycles, ribosomal proteins, and proteins related to transport and secretion. Controlling inverted repeat of chaperone expression and inverted repeat DNA binding sequences, as well as regions recognized by sigma factor 32, elements involved in the genetic regulation of the bacterial stress response, were identified in the promoter regions of several of the genes coding proteins, involved in the C. violaceum stress response. We found that 30 °C is the optimal growth temperature for C. violaceum, whereas 25, 35, and 40 °C are stressful temperatures that trigger the expression of chaperones, superoxide dismutase, a probable small heat shock protein, a probable phasing, ferrichrome-iron receptor protein, elongation factor P, and an ornithine carbamoyltransferase catabolite. This information improves our comprehension of the mechanisms involved in stress adaptation by C. violaceum.

Hydride generation using a metallic atomizer after microwave-assisted extraction for inorganic arsenic speciation in biological samples.

The present speciation method reports the determination of inorganic arsenic forms, using metallic furnace hydride generation atomic absorption spectrometry. The inorganic As speciation is carried out using mild conditions for hydride formation, such as slightly acid pH media (4.50) and low tetrahydridoborate(1-) concentration (0.1% (w/v)). Limits of detection and quantification of 2.0 and 6.6 µg L(-1) of iAs(III) are obtained using optimized conditions. Additionally, microwave-assisted extraction using water as solvent is carried out to provide the appropriate environment for As species extraction as well as impeding inter-conversion between species. With these analytical strategies, As was accurately determined (at 99.9% confidence level) in water and plankton samples.

Image analysis of two-dimensional gel electrophoresis for comparative proteomics of transgenic and non-transgenic soybean seeds.

Considering the importance of bidimensional electrophoresis and image analysis in comparative proteomics, the parameters that influence the analysis of protein expression of transgenic and non-transgenic soybean seeds were evaluated. The loaded mass of the proteins (150-500 microg), the pH separation range (3-10 or 4-7), and manual/automatic image editing were evaluated. Additionally, after optimizing the conditions, histograms and matchings were obtained in order to accurately analyze the variations (90%) in protein expression. From this, 10 proteins displayed significant differences in expression, and eight of them were characterized and identified by mass spectrometry.

Molecularly imprinted polymer as a solid phase extractor in flow analysis.

Molecularly imprinted polymers (MIPs) are novel alternative materials for solid phase extraction. Applications in flow analysis are recent and enhanced in-line separation/concentration procedures have been proposed. Use of flow systems is very important in the context. The aim of this review is then to highlight the implementation of MIP as solid phase extractor in flow analysis, emphasizing potentialities, limitations and applications.

Grape bagasse as an alternative natural adsorbent of cadmium and lead for effluent treatment.

This work investigated the utilization of grape bagasse as an alternative natural adsorbent to remove Cd(II) and Pb(II) ions from laboratory effluent. X-ray diffractometry, Fourier transform infrared spectroscopy, scanning electron microscopy, nuclear magnetic resonance, thermogravimetric analyses, surface analysis, porosity and porous size were used for characterization of the material. Batch experiments were carried out to evaluate the adsorption capacity of the material. Parameters such as adsorption pH and contact time were optimized for the maximum accumulation onto the solid surface. The pH values found were 7 and 3 for Cd(II) and Pb(II), respectively, and contact time was 5 min for both metals. Adsorption capacity for metals were calculated from adsorption isotherms by applying the Langmüir model and found to be 0.774 and 0.428 mmol g(-1) for Cd(II) and Pb(II), respectively. The competition between metals for the same adsorption sites on grape bagasse was also evaluated, showing an increasing affinity for Pb(II) over Cd(II) when only these metals are present. The potential of this material was demonstrated by efficient metal removal from laboratory effluent using a glass column. The results indicate that the referred material could be employed as adsorbent for effluent treatment, especially due to its easy acquisition and low cost as well as the fast adsorption involved.

Grape bagasse as a potential biosorbent of metals in effluent treatments.

Grape bagasse generated in the wine production process was characterized through X-ray diffractometry, Fourier transform infrared spectroscopy, scanning electron microscopy, nuclear magnetic resonance and thermogravimetric analysis. The efficiency of this natural material for Cd(II) and Pb(II) adsorption was evaluated using a batch adsorption technique. Factors affecting metal adsorption such as pH and contact time were investigated. Maximum adsorption was found to occur at pH 7.0 and 3.0 for Cd(II) and Pb(II), respectively, and a contact time of 5min was required to reach equilibrium for both metals. With these conditions, adsorption studies were performed using a single solution. In addition, to calculate the adsorption capacities for each metal, the Langmüir isotherm model was used. The adsorption capacities were found to be 0.479 and 0.204mmolg(-1) for Cd(II) and Pb(II), respectively. The results showed that grape bagasse could be employed as a low-cost alternative adsorbent for effluent treatment.

Metal-binding proteins scanning and determination by combining gel electrophoresis, synchrotron radiation X-ray fluorescence and atomic spectrometry.

In the present work, protein bands from in vitro embriogenic callus (Citrus sinensis L. Osbeck) were investigated using micro-synchrotron radiation X-ray fluorescence (muSR-XRF) after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) separation. Metal-binding protein quantification was done after microwave oven decomposition of gel by synchrotron radiation total reflection X-ray fluorescence (SR-TXRF), flame atomic absorption spectrometry (FAAS) and flame atomic emission spectrometry (FAES). According to the analysis of the protein bands, it is possible to observe that both 81 and ca. 14 kDa proteins present different Fe signal intensity at different positions. The analysis of 53 kDa protein, showed even more interesting results. Besides Fe, the muSR-XRF experiments indicate the presence of Ca, Cu, K and Zn. Chemical elements such as Cu, K, Fe and Zn were determined by SR-TXRF, Mg by FAAS and Na by FAES. Ca was determined by SR-TXRF and FAAS only for accuracy check. In the mineralised protein bands of 81 and around 14 kDa band, only Fe was determined (105 and 21.8 microg g(-1)). For those protein bands (86-ca. 14 kDa) were determined, Ca, K, Cu and Zn in a wide concentration range (42.4-283, 2.47-96.8, 0.91-15.9 and 3.39-29.7 microg g(-1), respectively).

Determination of Cd and Pb in food slurries by GFAAS using cryogenic grinding for sample preparation.

A simple method combining slurry sampling after cryogenic grinding and the use of a permanent modification of the integrated platform inside the transversely heated graphite atomizer (THGA) was proposed for the determination of Cd and Pb in foods. Potentialities of the cryogenic grinding were evaluated for grinding different materials of difficult homogenization such as high fat and high fiber tissues. Animal and vegetal samples were cut into small pieces and ground in liquid nitrogen for 2 min. Slurries were prepared directly in the autosampler cup after cryogenic grinding by transferring an exact amount of homogeneous powdered material (5-20 mg) to the cup, followed by 1.00 mL of 0.2% (v/v) HNO3 containing 0.04% (v/v) Triton X-100 and sonication for 30 s, before transferring into the platform previously coated with 250 microg W and 200 microg Rh. Use of a tungsten carbide-rhodium permanent modifier combined with NH4H2PO4 conventional modifier improves tube lifetime and increases the pyrolysis temperature for Cd. Homogeneity tests, carried out by comparing the between- and within-batch precision for each kind of sample, showed no significant differences at the 95% confidence level, indicating good homogeneity for 5-20 mg masses. Detection limits were 3.3 ng g(-1) Cd and 75 ng g(-1) Pb for 1% m/v slurries. Results for determination of Cd and Pb in foods slurries were in agreement with those obtained with digested samples, since no statistical differences were found by the paired t-test at the 95% level.