A new application of humic substances: Activation of supports for invertase immobilization

Invertase was immobilized on aminopropyl silica (APTS-SiO2) activated with humic substances (APTS-SiO2-HS) and on aminopropyl silica activated with glutaraldehyde (APTS-SiO2-GA). The resulting activity of both systems was compared. Humic substances (HS) used for the activation of the silica were extracted from soil of Cananéia, São Paulo State, Brazil, according to the procedure recommended by the International Humic Substances Society. Activity was determined by measuring the rate of formation of reduced sugars using the reaction with dinitrosalicylic acid (DNS). The amount of HS bound on the APTS-SiO2 was equal to 50 mg. The maximum amount of invertase immobilized on APTS-SiO2-HS was 15200 U/g while in the system APTS-SiO2-GA it was 13400 U/g. The experimental enzymatic activity was 3700 and 3300 U/g, for the systems APTS-SiO2-HS and APTS-SiO2-GA, respectively. Considering the increased amount and activity of immobilized enzyme compared with the glutaraldehyde method, it was concluded that this technique opens a new perspective in the preparation of supports for enzyme immobilization employing humic substances. © Springer-Verlag 2000.

Toxic mechanism of nickel exposure on cardiac tissue

The incidence of cardiovascular disease has increased in the general population, and cardiac damage is indicated as one important cause of mortality. In addition, pollution and metal exposure have increased in recent years. For this reason, toxic effects of metals, such as nickel, and their relation to cardiac damage should be urgently established. Although free radical-mediated cellular damage and reactive oxygen species have been theorized as contributing to the nickel mechanism of toxicity, recent investigations have established that free radicals may be important contributors to cardiac dysfunction. However, there is little information on the effect of nickel exposure on markers of oxidative stress in cardiac tissue. Nickel exposure (Ni2+ 100 mg L-1 from NiSO4) significantly increased lipoperoxide and total lipid concentrations in cardiac tissue. We also observed increased serum levels of cholesterol (59%), lactate dehydrogenase (LDH-64%), and alanine transaminase (ALT-30%) in study animals. The biochemical parameters recovered to the control values with tocopherol intake (0.2 mg 200 g-1). Vitamin E alone significantly decreased the lipoperoxide concentration and increased superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities in the heart. Since no alterations were observed in catalase and GSH-Px activities by nickel exposure while SOD activities were decreased, we conclude that superoxide radical (O2 -) generated by nickel exposure is of primary importance in the pathogenesis of cardiac damage. Tocopherol, by its antioxidant activity, decreased the toxic effects of nickel exposure on heart of rats.

Effect of hydrated sodium calcium aluminosilicate on the prevention of the toxic effects of ochratoxin

We studied the efficacy of hydrated sodium calcium aluminosilicate (HSCAS) as an inhibitor of the toxic effects of ochratoxin in broilers from 1 to 42 d of age. A total of 288 broilers was distributed into four treatments with four replicates of 18 birds each: T1, control; T2, 0.25% HSCAS; T3, 2 ppm of ochratoxin; T4, 0.25% HSCAS + 2 ppm ochratoxin. The parameters evaluated were feed intake; weight gain; feed conversion; relative weights of the liver, kidneys, and bursa; and serum levels of Ca, P, total protein (TP), aspartate aminotransferase (AST) and γ-glutamiltransferase (GGT). Ochratoxin in the diet negatively affected (P < 0.05) all performance parameters evaluated when the birds were 21 and 42 d of age. However, HSCAS did not affect performance, and there was no interaction between HSCAS and dietary ochratoxin. The liver and the kidneys of birds fed ochratoxin with or without HSCAS were relatively heavier (P < 0.05) than those of the control birds, demonstrating the influence of ochratoxin, but not of HSCAS, on the relative weight of these organs. Although the bursa of birds exposed to ochratoxin with or without HSCAS had a lower relative weight as compared to control birds, the difference was not significant. Ca, P, and TP serum levels were lower (P < 0.05) in birds exposed to ochratoxin, whereas AST and GGT levels were higher (P < 0.05) in these birds. These results reflect that ochratoxin in the diet impaired the productivity indexes and that HSCAS did not improve these parameters.

Avaliação da atividade mutagênica do resíduo sólido de uma fábrica de confecções de meias e lingerie em ensaios com Salmonella typhimurium

Cancer is one of the most hazardous effects to human health caused by the exposition to chemical agents. The search for new technological solutions in the industrial field led to a rapid increase in the productive sector, causing the workers to be exposed to millions of potentially toxic agents, substances potentially harmful to health. This study presents the mutagenic activity of sweepings from a sock and lingerie factory in Araraquara-Brazil, assayed with Salmonella typhimurium. All the extracts from the factory had mutagenic on activity the YG1024 strain, which is extremely sensitive to detect the mutagenic activity of the arilhydroxilamines, nitroarenes and aromatic amines. The extracts were non-mutagenics for the strains TA100 and TA98. The analysis of the mutagenicity of industrial residues is highly important because employees that participate in the production are directly exposed to those agents, as well as to the environment where the garbage is deposited.

Competition between humic substances and α-amino acids by metal species

The aim of the present work was to carry out experimental comparison between humic substances (HS) and representative α-amino acids (methionine, methionine sulfoxide and cysteine hydrochloride) in relation to the complexation of biologically active trace elements (Al, Cu, Pb, Mn, Zn, Cd and Ni). A mobile time-controlled tangential-flow UF technique was applied to differentiate between HS-metal and α-aminoacids-metal complexes. Metal determinations were conventionally carried out using a ICP-OES. The results showed that HS may be considered as a selective complexing agents with higher metal bonding capability in relation to Al, Cu and Pb, the fact that may be clinically important.

Method for the evaluation of insecticidal activity over time in Atta sexdens rubropilosa workers (Hymenoptera: Formicidae)

The active ingredients used in the formulation of toxic baits for leaf-cutting ants (Hymenoptera: Formicidae) should possess a delayed action defined as an insecticidal activity whereby worker mortality is ≤15% at 24 hours and ≥90% at 21 days. Serious shortcomings have occurred in the search for new active ingredients, such as the initial selection of fenoxycarb, copper oxychloride and diflubenzuron, compounds considered very promising but whose inefficiency was verified only later, indicating methodological problems. In view of this situation, we developed a classification method for insecticidal activity over time using workers of the leaf-cutting ant Atta sexdens rubropilosa Forel. The insecticides used were fipronil, sulfluramid GX071HB and sulfluramid GX439, vehicled in an attractive pasty formulation prepared based on citrus pulp. The results obtained were consistent from a toxicological viewpoint and agreed with the literature in terms of the control of colonies. Sulfluramids were found to possess a delayed action at a broad range of concentrations, in agreement with the fact that these substances are highly effective in the control of all leaf-cutting ant species. The smaller range of concentrations of fipronil with delayed action is probably related to its lower efficacy for species more difficult to control such as Atta capiguara (Forti et al. 2003). We discuss the importance of relating behavioral particularities to the specific feeding habits of leaf-cutting ants, with methodological adequacy of the assessment of insecticides aimed at toxic baits.

Studies on the interaction between humic substances and conducting polymers for sensor application

The interaction between humic substances and poly(o-ethoxyaniline) (POEA), a conducting polymer, was investigated for both solution and self-assembled films. The results have shown that the humic substances induce a doping of POEA by protonation, as indicated by UV-Vis and Raman spectroscopies. The atomic force microscopy (AFM) studies on the self-assembled films have shown that the average roughness of the polymer film has increased after exposing it to humic substances (fulvic and humic acids), consistent with the interaction between POEA and humic substances. However, this change in morphology is reversible by washing the films with water in agreement with the electrical data allowing using this system in sensor applications. Here, the sensor formed by an array of different sensing units was able to detect and distinguish humic substances in aqueous solution, as shown by multivariate analysis (principal component analysis). The motivation to detect humic substance comes due to its importance in terms of quality control of water or soil. ©2005 Sociedade Brasileira de Química.

Toxic and genotoxic effects of trivalent and hexavalent chromium - A review

During the last years, the emission of heavy metals to the environment has increased, causing a severe negative impact to the ecosystems and seriously compromising human health due to their mutagenic potential. Tri- (III) and hexavalent (VI) chromium (Cr) constitute the oxidative states of the metal chromium that are active in living organisms. These two oxidation states of the chromium differ with regards to their cellular effects, mainly due to the different abilities they possess in relation to easy of transport through biological membranes. Cr VI is transported into the cell through transference channels of endogenous anions that are isostructural and isoelectronical to Cr VI, such as SO 4 -2 and HPO 4 -2. On the other hand, Cr III is unable to diffuse through the cell membrane. Its existence inside the cells is generally due to the reduction of Cr VI, the endocytosis, or the absortion by the cells via phagocytosis. Cr III acts directly on the DNA molecule, while Cr VI reacts little with this molecule. In the ecosystem, however, Cr VI is more dangerous since this is the form that presents greater reactivity with biological membranes, crossing them and being easily incorporated into the cell. In the cell it is biotransformed to Cr III, a potentially mutagenic molecule. In vivo and in vitro studies have shown that organisms exposed to Cr VI present greater induction to a variety of damages to the DNA molecule. Among the damages induced by Cr, changes in the structure of the DNA molecule have been reported, with breaks of the major chain and base oxidation. In the organisms, these alterations generate chromosomal aberrations, micronucleus formation, sister chromatid exchanges, and errors in DNA synthesis.

The role of workers in transferring queen substances and the differences between worker castes in the leaf-cutting ant, Acromyrmex subterraneus brunneus (Hymenoptera: Formicidae)

The present study focused on the relationship between the queen and workers in Acromyrmex subterraneus brunneus colonies mediated by the possible transfer of substances from the fertile to the sterile caste. The queens were submitted to different situations of physical limitation, i.e., they were kept isolated in cages with holes that only permitted the entry of workers but left the queen isolated. A tracer (water-soluble dye) was applied to the pronotum and gaster of the queen and its dispersal among workers was analyzed. The results demonstrated that the minor sub caste (0.7-0.9 mm) passed on the dye through allogrooming and self-grooming, or transferred the dye through trophallaxis to the major sub caste (1.2-2.0 mm) when they were not in direct contact with the queen. These findings indicate the communication and probable transfer of substances from the queen to the workers, as well as a substance transfer between workers.

Detection of enterotoxin and toxic shock syndrome toxin 1 genes in Staphylococcus, with emphasis on coagulase-negative staphylococci

The detection of staphylococcal enterotoxins is decisive for the confirmation of an outbreak and for the determination of the enterotoxigenicity of strains. Since the recognition of their antigenicity, a large number of serological methods for the detection of enterotoxins in food and culture media have been proposed. Since immunological methods require detectable amounts of toxin, molecular biology techniques represent important tools in the microbiology laboratory. In the present study, polymerase chain reaction (PCR) was used to identify genes responsible for the production of enterotoxins and toxic shock syndrome toxin 1 (TSST-1) in S. aureus and coagulase-negative staphylococci (CNS) isolated from patients and the results were compared with those obtained by the reverse passive latex agglutination (RPLA) assay. PCR detection of toxin genes revealed a higher percentage of toxigenic S. aureus strains (46.7%) than the RPLA method (38.3%). Analysis of the toxigenic profile of CNS strains showed that 26.7% of the isolates produced some type of toxin, and one or more toxin-specific genes were detected in 40% of the isolates. These results suggests the need for further studies in order to better characterize the pathogenic potential of CNS and indicate that attention should be paid to the toxigenic capacity of this group of microorganisms.

The influence of seasonalness on the structural characteristics of aquatic humic substances extracted from Negro River (Amazon state) waters: Interactions with Hg(II)

In this work, humic substances were extracted from water samples collected monthly from the Negro River basin in the Amazon state (Brazil) to study their properties in the Amazonian environment and interactions with the mercury ion considering the influence of seasonalness in this formation. The C/H, C/N and C/O atomic ratio parameters, functional groups, concentration of semiquinone-type free radicals, pH, pluviometric and fluviometric indices, and mercury concentrations were interpreted using hierarchical cluster analysis (HCA) and principal component analysis (PCA). The statistical analyses showed that when the pluviometric index was greater and the fluviometric index was smaller, the degree of humification of aquatic substances was greater. The following decreasing order of the degree of humification of the AHS collected monthly was established: Nov/02 to Feb/03 > Mar/02 to May/02 > Jun/02 to Oct/02. The greatest concentrations of mercury were detected in more humidified samples. These results suggest that due to inter and/or intra-molecular rearrangements, the degree of humification of aquatic humic substances is related to its affinity for Hg(II) ions. ©2007 Sociedade Brasileira de Química.

Separation of the toxic zierin from Zollernia ilicifolia by high speed countercurrent chromatography

Preliminary pharmacological assays of the 70% methanol extract from the leaves of the Brazilian medicinal plant Zollernia ilicifolia Vog. (Fabaceae) showed analgesic and antiulcerogenic effects. Previous analyses have shown that this extract contains, besides flavonoid glycosides and saponins, a toxic cyanogenic glycoside. Flavonoids and saponins are compounds reported in literature with antiulcerogenic activity. In this work, we developed a methodology to separate the cyanogenic glycoside from these compounds in order to obtain enough amount of material to perform pharmacological assays. The cyanogenic glycoside zierin (2S)-β-D-glucopyranosyloxy-(3-hydroxy-phenyl)- acetonitrile was separated from the other components by high speed countercurrent chromatography (HSCCC). The solvent system used was composed of chloroform-methanol-n-propanol-water (5:6:1:4, v/v/v/v). This technique led to the separation of zierin from the possible active compounds of Zollernia ilicifolia.

Potencial tóxico dos alginatos para uso odontológico

Alginate or irreversible hydrocolloid is one the most accepted and frequently employed impression materials in dental practice. Substances like zinc, cadmium, lead silicate and fluorides, which are included in several alginate brands with the aim of improving their physical, chemical and mechanical properties, are a source of serious concern as regards their toxicity. Some brands of alginate have been reported to contain potentially toxic fluorides and metals such as cadmium, lead and zinc silicates, either singly or combined. Consequently, special care should be taken while preparing of these materials. It is necessary to monitor potentially toxic chemicals and metals in the alginates continually to avoid contamination of dental professionals and patients. In this review, alginates used in dentistry are analyzed for potential toxicity.

Solvent capacity of different substances on gutta-percha and resilon

The aim of this study was to evaluate the effectiveness of 3 solvents (eucalyptol, orange oil, and xylol) on 2 types of gutta-percha (conventional and thermoplastic) and Resilon. Specimens (10 mm diameter × 1 mm thick; n=7 per condition) were prepared and maintained at 37°C for 48 h. Each specimen was weighed on a precision scale every 24 h until its mass was stable, at which time the initial mass was determined. Specimens (n=7) were then immersed in the solvent solutions and, after 48 h at 37°C, they were reweighed at 24- h intervals, until stabilization (final mass). The difference between the final and the initial mass determined the solvent capacity of each solvent. Data were subjected to analysis of variance and Tukey's test at 5% significance level. The results demonstrated that xylol was the most effective, especially on conventional gutta-percha and Resilon (p<0.05). Eucalyptol and orange oil were more effective on thermoplastic gutta-percha than the other materials (p<0.05). It was concluded that all evaluated substances presented solvent action, but xylol was the most effective on both gutta-percha or Resilon.

Alterations in the fat body cells of rhinocricus padbergi (Diplopoda) resulting from exposure to substrate containing sewage sludge

Abstract The final disposal of residues generated at sewage treatment plants (STPs) has become a major problem for cities, due to the increase in the amount of treated sewage. One of the alternatives for the residue, labeled sewage sludge, is its reuse in agriculture and in degraded soil. However, not all pathogens and metals present in it are eliminated during treatment. Diplopods have been used as bioindicators in ecotoxicological tests as they are constantly in close contact with the soil. Owing to this fact, the purpose of this study was to expose specimens of the diplopod Rhinocricus padbergi to substrate containing sewage sludge collected at STPs to analyze morphological alterations in their parietal and perivisceral fat body, where substances are stored. The exposures were held for 7, 15, or 90 days at different concentrations of sewage sludge (control, 1%, 10%, and 50%). The parietal fat body showed no alterations in any of the three exposure periods or concentrations. Alterations in the perivisceral fat body were observed for all exposure periods. According to the results, we suggest that the sludge used has toxic agents responsible for changing the animal's perivisceral fat body. © 2012 Microscopy Society of America.