Sabrina Manente

A five-year study on the heavy-metal pollution of Guanabara Bay sediments (Rio de Janeiro, Brazil) and evaluation of the metal bioavailability by means of geochemical speciation

Surface sediments of the Guanabara Bay (Rio de Janeiro, Brazil) were analyzed by a sequential extraction procedure for Cd, Cr, Cu, Pb, Zn, Mn and Fe, determining their distribution among five geochemical phases and in the nitric acid extractable phase. Bioavailable phases and non-bioavailable phases have been determined in six transects in the bay to define the significant level of pollution due to sediment metal contamination. A multiple correlation showed limited responsibility of Mn and Fe oxides and humic acids in the adsorption process, which allowed discrimination among the different processes and suggested the strong influence of the hydrogen sulfide present in the highly reduced bay bottom environment. The authors suggest the need to avoid disturbing bottom sediment by dredging or by artificial bottom aeration which could result in a rapid worsening of the environment due to the accelerated formation of more soluble oxygenated metal compounds making the toxic metals much more available to the benthic fauna and to the bay biota in general.

Heavy metal pollution in central Venice lagoon bottom sediments: evaluation of the metal bioavailability by geochemical speciation procedure.

Bottom sediments of the central area of the Venice Lagoon (Italy) and some lagoon canals, chosen as an anaerobic reference environment, were analyzed by a sequential extraction procedure for Cr, Cu, Ni, Pb, Zn, Mn, and Fe. Bio-Available Phases (BAPs) and Non-Bio-Available Phases (NBAPs) have been determined in the Malamocco-Marghera Canal, to define the significant level of heavy metal contamination of the sediment. Relationships among metal distributions with Fe/Mn oxides/hydroxides and sulfides were determined in both environments, using statistical methods. The central lagoon area shows low pollution and a balanced influence of all the sediment components in the metal complexing processes, mostly due to the manganese and iron matrices. The inner canals are very polluted with heavy metals strongly bonded to H2S that form highly insoluble compounds, as demostrated through the correlation analysis. The authors suggest different procedures for lagoon restoration due to the different sediment behaviours, i....

AN IN VITRO STUDY ON THE TOXIC EFFECTS OF NONYLPHENOLS (NP) IN MITOCHONDRIA

This paper is focused on alkylphenols, compounds which are formed by the biodegradation of polyethoxilatedalkylphenols detergents. Our experiments show that alkylphenols act not only as detergents, but also as uncouplers of the oxidative phosphorylation. This effect, can be observed at very low doses, thus suggesting that the preferential target of nonylphenols in living organisms are mitochondria.

Methylmercury induces the opening of the permeability transition pore in rat liver mitochondria.

Interactions of methylmercury (CH(3)HgCl) with non-energized mitochondria from rat liver (non-respiring mitochondria) have been investigated in this paper. It has been shown that CH(3)HgCl induces swelling in mitochondria suspended in a sucrose medium. Swelling has also been induced by detergent compounds and by phenylarsine, a chemical compound which induces opening of the permeant transition pore (MTP). Opening of the MTP is inhibited by means of cyclosporine A. Results indicate that the swelling induced by CH(3)HgCl, as in the case of phenylarsine, is inhibited by cyclosporine A and Mg(2+), while swelling induced by detergent compounds is not cyclosporine sensitive. This comparison suggests that CH(3)HgCl induces opening of a permeability transition pore (MTP). Since the opening of an MTP induces cell death, this interaction with MTP could be one of the causes of toxicity of CH(3)HgCl.

The interactions of cobalt(II) with mitochondria from rat liver

The interactions of Co2+ with mitochondria have been investigated. The results indicate that Co2+ inhibits ATP synthesis. Further investigations into ATP synthesis mechanisms indicated that inhibition is due to the opening of a transmembrane pore. The opening of this pore causes the collapse of the high-energy intermediate where, under a pH and a potential gradient, the energy is stored and subsequently utilized to form ATP from ADP.

Heparin-induced structural and functional alterations of bovine trypsin

To investigate the mechanism whereby heparin can modulate the activity of serine proteinases, bovine trypsin was chosen as reference and treated with heparin at 10, 100 and 200 micrograms/ml, in buffer solvents, with and without incubation at 37 degrees C. Heparin caused rapid, buffer- and pH-dependent decrease in trypsin solubility due to the generation of insoluble fragments from proteinase. Desalting treatments variously restored solubility by removing insoluble material. UV absorption and fluorescence emission spectra revealed significant heparin-induced conformational alterations in the trypsin molecule, the maximal effect being apparent at a proteinase-to-heparin molar ratio ranging from 1.6 to 1.0. The involvement of the catalytic sites of trypsin by heparin was further confirmed by the significant reduction in the difference absorption spectra of proflavine. Both proteolytic and esterolytic activities of trypsin were shown to be markedly decreased by heparin, especially after 5 h incubation at 37 degrees C. However, when the proteolytic and esterolytic activities of trypsin were measured on fresh solutions not submitted to desalting treatments, variable activation instead of inhibition of both activities was observed in the presence of heparin, this effect waning spontaneously in time or after desalting treatment. The paradoxical increase in functional activities was not inhibited by soybean trypsin inhibitor and was accompanied by denaturation and fragmentation of the proteinase as demonstrated by spectroscopic analyses and SDS-PAGE of fresh solutions. The results obtained indicated that heparin causes a rapid, time- and temperature-dependent conformational alteration of trypsin with irreversible denaturation and degradation of the proteinase. The underlying mechanism appears to be heparin-catalyzed oxidative degradation of trypsin due to liberation of oxygen radicals which are also responsible for the temporary increase in catalytic functions.

Bidirectional fluxes of spermine across the mitochondrial membrane

The polyamine spermine is transported into the mitochondrial matrix by an electrophoretic mechanism having as driving force the negative electrical membrane potential (ΔΨ). The presence of phosphate increases spermine uptake by reducing ΔpH and enhancing ΔΨ. The transport system is a specific uniporter constituted by a protein channel exhibiting two asymmetric energy barriers with the spermine binding site located in the energy well between the two barriers. Although spermine transport is electrophoretic in origin, its accumulation does not follow the Nernst equation for the presence of an efflux pathway. Spermine efflux may be induced by different agents, such as FCCP, antimycin A and mersalyl, able to completely or partially reduce the ΔΨ value and, consequently, suppress or weaken the force necessary to maintain spermine in the matrix. However this efflux may also take place in normal conditions when the electrophoretic accumulation of the polycationic polyamine induces a sufficient drop in ΔΨ able to trigger the efflux pathway. The release of the polyamine is most probably electroneutral in origin and can take place in exchange with protons or in symport with phosphate anion. The activity of both the uptake and efflux pathways induces a continuous cycling of spermine across the mitochondrial membrane, the rate of which may be prominent in imposing the concentrations of spermine in the inner and outer compartment. Thus, this event has a significant role on mitochondrial permeability transition modulation and consequently on the triggering of intrinsic apoptosis.

The interaction of tributyllead with lysosomes from rat liver

The interactions of tributyllead with lysosomes from rat liver have been studied. It results that the organometal compound induces a fast alkalinization in energized lysosomes. The interpretation is that the compound is a potent proton carrier. This function could explain the toxicity, in particular at neurological level of the compound.

A comparison between the responses of neutral red and acridine orange: Acridine orange should be preferential and alternative to neutral red as a dye for the monitoring of contaminants by means of biological sensors

The acridine orange (AO) and neutral red (NR) dyes, commonly used as probes to measure the internal pH in acidic vesicles, are compared in this article. The comparison between the two dyes (arising from calculations taking into account their analytical constants) illustrated that the use of AO is preferential to that of NR because the AO response is sensitive over the whole pH range between 4.0 and 7.4, whereas the NR response is effective only between pHs 4.0 and 6.0. In addition, it became evident from the mitochondrial respiration response that NR, unlike AO, is a protonophore. When taken into consideration, these two properties suggest that AO is more suitable than NR as an indicator of toxicity measurements in water samples because the environmental toxic compounds induce pH changes in the acidic vesicles of biological structures that are used as environmental biosensors.

Milestones and recent discoveries on cell death mediated by mitochondria and their interactions with biologically active amines

Mitochondria represent cell “powerhouses,” being involved in energy transduction from the electrochemical gradient to ATP synthesis. The morphology of their cell types may change, according to various metabolic processes or osmotic pressure. A new morphology of the inner membrane and mitochondrial cristae, significantly different from the previous one, has been proposed for the inner membrane and mitochondrial cristae, based on the technique of electron tomography. Mitochondrial Ca2+ transport (the transporter has been isolated) generates reactive oxygen species and induces the mitochondrial permeability transition of both inner and outer mitochondrial membranes, leading to induction of necrosis and apoptosis. In the mitochondria of several cell types (liver, kidney, and heart), mitochondrial oxidative stress is an essential step in the induction of cell death, although not in brain, in which the phenomenon is caused by a different mechanism. Mitochondrial permeability transition drives both apoptosis and necrosis, whereas mitochondrial outer membrane permeability is characteristic of apoptosis. Adenine nucleotide translocase remains the most important component involved in membrane permeability, with the opening of the transition pore, although other proteins, such as ATP synthase or phosphate carriers, have been proposed. Intrinsic cell death is triggered by the release from mitochondria of proteic factors, such as cytochrome c, apoptosis inducing factor, and Smac/DIABLO, with the activation of caspases upon mitochondrial permeability transition or mitochondrial outer membrane permeability induction. Mitochondrial permeability transition induces the permeability of the inner membrane in sites in contact with the outer membrane; mitochondrial outer membrane permeability forms channels on the outer membrane by means of various stimuli involving Bcl-2 family proteins. The biologically active amines, spermine, and agmatine, have specific functions on mitochondria which distinguish them from other amines. Enzymatic oxidative deamination of spermine by amine oxidases in tumor cells may produce reactive oxygen species, leading to transition pore opening and apoptosis. This process could be exploited as a new therapeutic strategy to combat cancer.