Cláudio Nahum Alves

Evaluation of total mercury concentrations in fish consumed in the municipality of Itaituba, Tapajós River Basin, Pará, Brazil.

The environmental exposure to mercury by riverine and Indian communities can occur through the ingestion of fish and products contaminated by mercury compounds. The present study aims to evaluate the total mercury concentration in the different fish species most consumed in the municipality of Itaituba, Tapajós river basin, where there are intense gold-mining activities. These fish samples were analyzed by atomic absorption spectrophotometry by cold vapor technique, Automatic Mercury Analyzer HG-3500. The largest mercury concentrations were found in the carnivorous species ranging from 112.4 to 2250 microg/g, while the detritivorous, herbivorous and omnivorous species presented total mercury levels ranging from 3.2 to 309.8 microg/g, which is below the limit established by the World Health Organization. This paper also reports and identifies which species are more polluted and present a statistical relationship among concentration and weight of the carnivorous species studied in detail, Brachyplatystoma flavicans (dourada) (r2 = 0.691) and Pseudoplatystoma sp. (surubim) (r2 = 0.654).

Application of Aqai Stalks as Biosorbents for the Removal of the Dye Procion Blue MX-R from Aqueous Solution

The aqai palm stalk (Euterpe oleracea) is a food residue used in its natural form (AS) and also protonated (AAS) as biosorbents for the removal of the textile dye Procion Blue MX-R from aqueous solutions. This biosorbent was characterized by infrared spectroscopy, scanning electron microscopy, and nitrogen adsorption-desorption curves. The effects of pH, biosorbent dosages, and shaking time on the biosorption capacities were studied. In the acidic pH region (pH 2.0) the biosorption of the dye was favorable. The minimum contact time to obtain the equilibrium was 8 hours for AS and AAS biosorbents. The general order kinetic model provided the best fit to the experimental data compared with the pseudo-first order and pseudo-second order kinetic adsorption models. The equilibrium data were fitted to the Langmuir, Freundlich, and Sips isotherm models. For both dyes the equilibrium data were best fitted to the Sips isotherm model. The enthalpy and entropy of adsorption of PB were obtained from adsorption equilibrium experiments ranging from 298 to 323 K. Simulated dyehouse effluent was used to check the applicability of the proposed adsorbents for effluent treatment.

Structure of Dihydrochalcones and Related Derivatives and Their Scavenging and Antioxidant Activity against Oxygen and Nitrogen Radical Species

Quantum mechanical calculations at B3LYP/6-31G** level of theory were employed to obtain energy (E), ionization potential (IP), bond dissociation enthalpy (O-H BDE) and stabilization energies (ΔEiso) in order to infer the scavenging activity of dihydrochalcones (DHC) and structurally related compounds. Spin density calculations were also performed for the proposed antioxidant activity mechanism of 2,4,6-trihydroxyacetophenone (2,4,6-THA). The unpaired electron formed by the hydrogen abstraction from the phenolic hydroxyl group of 2,4,6-THA is localized on the phenolic oxygen at 2, 6, and 4 positions, the C3 and C6 carbon atoms at ortho positions, and the C5 carbon atom at para position. The lowest phenolic oxygen contribution corresponded to the highest scavenging activity value. It was found that antioxidant activity depends on the presence of a hydroxyl at the C2 and C4 positions and that there is a correlation between IP and O-H BDE and peroxynitrite scavenging activity and lipid peroxidation. These results identified the pharmacophore group for DHC.

Kojic acid, a secondary metabolite from Aspergillus sp., acts as an inducer of macrophage activation.

KA (kojic acid) is a secondary metabolite isolated from Aspergillus fungi that has demonstrated skin whitening, antioxidant and antitumour properties among others. However, limited information is available regarding its effects on macrophages, the major cell involved in cell defence. The aim of the present study was to analyse whether KA affects functional properties related to macrophage activation, such as phagocytosis and spreading ability over a substrate. Treatment of resident macrophages with 50 μg/ml KA for 1 h induced both morphological and physiological alterations in cells. Immunofluorescence microscopy revealed enhanced cell spreading and an increase in cell surface exposure, associated with a rearrangement of microtubules, actin filaments and intermediate filaments. KA also potentiated phagocytosis by macrophages, as demonstrated by the increase in phagocytic activity towards yeast, when compared to untreated cells. KA increased the production of ROS (reactive oxygen species), but not NO (nitric oxide) production. Three tests were used to assess cell viability; MTT [3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl tetrazolium bromide], NR (neutral red) uptake and PI (propidium iodide) exclusion test, which showed that macrophages maintain their viability following KA treatment. Results indicate that KA can modulate macrophage activation through cytoskeleton rearrangement, increase cell surface exposure, enhance the phagocytic process and ROS production. The study demonstrates a new role for KA as a macrophage activator.

A Quantum Mechanics/Molecular Mechanics Study of the Protein-Ligand Interaction of Two Potent Inhibitors of Human O-GlcNAcase : PUGNAc and NAG-Thiazoline

O-glycoprotein 2-acetamino-2-deoxy-beta- d-glucopyranosidase ( O-GlcNAcase) hydrolyzes 2-acetamido-2-deoxy-beta- d-glucopyranose ( O-GlcNAc) residues of serine/threonine residues of modified proteins. O-GlcNAc is present in many intracellular proteins and appears to have a role in the etiology of several diseases including cancer, Alzheimers disease, and type II diabetes. In this work, we have carried out molecular dynamics simulations using a hybrid quantum mechanics/molecular mechanics approach to determine the binding of two potent inhibitors, PUGNAc and NAG, with a bacterial O-GlcNAcase. The results of these simulations show that Asp-401, Asp-298, and Asp-297 residues play an important role in the protein-inhibitor interactions. These results might be useful to design compounds with more interesting inhibitory activity on the basis of its three-dimensional structure.

Mercury levels assessment in hair of riverside inhabitants of the Tapajós River, Pará State, Amazon, Brazil: Fish consumption as a possible route of exposure

The study present evaluated the levels of mercury (Hg) and methylmercury (MeHg) in hair samples of people from Barreiras community, riverside inhabitants of the Tapajós River (Pará, Brazil), an area impacted by clandestine gold mining, as well as we analyzed the levels of Hg and Se (selenium) in nine fish species (carnivores and non-carnivorous) from the Tapajós River, which stand out as the main species consumed by riverside inhabitants, to evaluate a relationship between frequency of fish consumption and Hg concentration, and also to evaluate possible mechanisms of fish protection (or non-protection) to Hg exposure by Se. Furthermore we analyze the water quality to evaluate the environmental trophic state, fact responsible by creating conditions that can potentiate the effects of toxic mercury. Concentrations of Hg and MeHg were analyzed in hair samples of 141 volunteers in different age band. Of those, 84.40% of samples present values above the threshold for biological tolerance, which is 6.00μgg(-1) of total Hg in hair. Total Hg, in men there was a variation of 2.07-24.93μgg(-1), while for women the variation was 4.84-27.02μgg(-1). Consequently, the level of MeHg in men presented a variation of 1.49-19.57μgg(-1), with an average of 11.68μgg(-1), while with women the variation was from 3.73 to 22.35μgg(-1), with an average of 10.38μgg(-1). In fish species, Hg concentrations in carnivorous species had an average of 0.66μgg(-1), higher than that permitted by current legislation, ranging from 0.30 to 0.98μgg(-1), while the non-carnivorous species have values below the recommended by the legislation averaging 0.09μgg(-1), ranging between 0.02 and 0.44μgg(-1). For Se in fish, show that among carnivores, the contents of Se ranged between 0.18 and 0.54μgg(-1) with a mean of 0.34μgg(-1), while for non-carnivores these values were of the order of 0.16-0.56μgg(-1), with an average of 0.32μgg(-1). In surface water quality variables at the sampling points all showed values in accordance with the range established by current legislation. In this regard, the results provided by this study, while not conclusive, are strong indicators that despite not having been shown the relationship between the concentration of mercury in hair and feeding habits along the Tapajós River basin communities showed that a plausible correlation exists between levels of mercury and selenium in fish. This fact may serve as a subsidy to research human health, because in the Amazon, there is still a lot to examine with regards to the full understanding of the Se cycle.

Avaliação do efeito antifúngico do óleo resina e do óleo essencial de copaíba (Copaifera multijuga Hayne)

Copaifera multijuga Hayne oil in natura and its fractions were evaluated as to their fungitoxic activities against five filamentous fungus species belonging to the genus Aspergillus and three yeast species of the genus Candida. Oleoresin and essential oil concentrations ranging from 0.08 mg mL-1 to 1.6 mg mL-1 were used for qualitative and quantitative analyses. The samples were placed onto 5mm-diameter paper discs distributed on Sabourauds medium in Petri dishes inoculated with spores of the microorganisms and incubated at 28oC for 10 days. A solution containing 1.6 mg mL-1 miconazole nitrate was used as positive control. Qualitative results showed that the oleoresin presented good activity, while a fraction of the essential oil was highly effective against Candida parapsilosis IOC-2882, Aspergillus flavus IOC-3874 and A tamarii IOC-187 with 16.0±1.4 mm, 19.5±2.1 mm and 12.5±3.5 mm inhibition halos, respectively. The quantitative evaluation showed that 0.3 mg mL-1 oleoresin inhibited the growth of Aspergillus flavus and Candida parapsilosis, while 0.08 mg mL-1 of the essential oil fraction reached this same activity.

Antifungal Activity and Computational Study of Constituents from Piper divaricatum Essential Oil against Fusarium Infection in Black Pepper

Fusarium disease causes considerable losses in the cultivation of Piper nigrum, the black pepper used in the culinary world. Brazil was the largest producer of black pepper, but in recent years has lost this hegemony, with a significant reduction in its production, due to the ravages produced by the Fusarium solani f. sp. piperis, the fungus which causes this disease. Scientific research seeks new alternatives for the control and the existence of other Piper species in the Brazilian Amazon, resistant to disease, are being considered in this context. The main constituents of the oil of Piper divaricatum are methyleugenol (75.0%) and eugenol (10.0%). The oil and these two main constituents were tested individually at concentrations of 0.25 to 2.5 mg/mL against F. solani f. sp. piperis, exhibiting strong antifungal index, from 18.0% to 100.0%. The 3D structure of the β-glucosidase from Fusarium solani f. sp. piperis, obtained by homology modeling, was used for molecular docking and molecular electrostatic potential calculations in order to determine the binding energy of the natural substrates glucose, methyleugenol and eugenol. The results showed that β-glucosidase (Asp45, Arg113, Lys146, Tyr193, Asp225, Trp226 and Leu99) residues play an important role in the interactions that occur between the protein-substrate and the engenol and methyleugenol inhibitors, justifying the antifungal action of these two phenylpropenes against Fusarium solani f. sp. piperis.

Quantum mechanical/molecular mechanical molecular dynamics simulation of wild-type and seven mutants of CpNagJ in complex with PUGNAc.

The enzyme O-glycoprotein 2-acetamino-2-deoxy-beta-d-glucopyranosidase (O-GlcNAcase) is responsible for the removal of N-acetylglucosamine moieties from 2-acetamido-2-deoxy-beta-D-glucopyranose (O-GlcNAc) residues of serine/threonine residues of modified proteins. We herein present results of hybrid quantum mechanics/molecular mechanics (QM/MM) molecular dynamics (MD) simulations applied to the study of the interactions established between a bacterial Clostridium perfringens homologue (CpNagJ) and PUGNAc, a potent known inhibitor of this enzyme. Electrostatic binding free energy and energy term decomposition have been computed for the wild-type CpNagJ and several mutants: D297N, D298N, Y335F, N390A, N396A, D401A, and W490A. The theoretical results have been compared with recently experimental data based on crystallographic and mutation studies on the same system. Our results reveal that, first, there is a strong interaction between Asp401, Asp298, and Asp297 residues and the PUGNAc inhibitor; and, second, the electrostatic substrate binding free energy is higher in wild-type, N390A and W490A mutants than in D297N, D298N, Y335F, N396A, and D401A ones, in accordance with the experimental results. Finally, both our theoretical predictions and the experimental data are compatible with a substrate-assisted reaction mechanism, involving two conserved aspartate residues.

A DFT Study of the Molecular Mechanisms of the Diels−Alder Reaction between Cyclopentadiene and 3-Phenyl-1-(2-pyridyl)-2-propen-1-one − Role of the Zn2+ Lewis Acid Catalyst and Water Solvent

The molecular mechanism of the Diels−Alder reaction between cyclopentadiene (1) and 3-phenyl-1-(2-pyridyl)-2-propen-1-one (2) in the absence and in the presence of a Zn2+ Lewis acid catalyst has been studied by quantum mechanical calculations at the B3LYP/6-31G* level of theory. A continuum model was selected to represent the effects of the water as solvent. For the uncatalyzed process, two channels, endo and exo, were characterized, and the mechanism corresponded to an asynchronous concerted reaction associated with a [4+2] process. The presence of a Lewis acid catalyst changed the mechanism drastically, the reaction taking place by a polar stepwise mechanism. In the first step, a C−C sigma bond was formed by the nucleophilic attack of 1 on the conjugate position of the Lewis acid coordinated α,β-unsaturated ketone to give a zwitterionic intermediate, while the second step was a ring-closure process by this intermediate to give the final formally [4+2] cycloadduct. The theoretical results have been compared with available experimental data and an understanding of the role of Lewis acids and water solvent emerges from analysis of the results. (© Wiley-VCH Verlag GmbH, 69451 Weinheim, Germany, 2002)