Simple and effective gas chromatographic mass spectrometric procedure for the speciation analysis of organotin compounds in specimens of marine mussels. An evaluation of the organotin pollution of the lagoon of venice.

This paper describes a simple, effective analytical procedure, based on a gas chromatographic mass spectrometric technique, for the speciation analysis of organotin compounds (OTC) in mussel samples. The direct alkylation reaction of the organotin chlorides in the aqueous digestion solution by NaBEt(4) allowed a short analysis time and a good recovery. The evaluation of the yield of each step constituting the analytical procedure indicated that the alkylation step is the most critical one. The proposed method was advantageously utilised to monitor the level of OTC pollution of the Lagoon of Venice. All the sites examined, both near to and far from anthropogenic activities, revealed significant levels of OTC pollution.

Effects of chronic dietary cadmium on hepatic glutathione levels and glutathione peroxidase activity in starlings (Sturnus vulgaris).

The effects of chronic exposure to dietary cadmium on the levels of hepatic glutathione (GSH) and on the activity of the glutathione peroxidase enzymes (GSH-Px) were studied for the first time in starlings (Sturnus vulgaris). Thirty-three individuals (17 females and 16 males) were divided into three groups: One represented the untreated control and two were respectively fed with diets containing 10 and 50 ppm cadmium chloride (CdCl(2)). The total duration of treatment was 22 weeks. The three groups respectively accumulated mean hepatic Cd residues of 2.29, 75.71, and 208.49 ppm. Hepatic GSH increased in the treated groups respectively 24% and 52% in comparison to controls. Total GSH-Px activity in the liver was inhibited in the group fed with 50 ppm, due to inhibition of the selenium-dependent fraction of the enzyme, while the selenium-independent fraction did not change significantly. During the treatment, after 14 weeks of exposure to cadmium, the 50 ppm-treated group showed a 47% decrease of the activity of the selenium-dependent GSH-Px and a 50% increase of the somatic liver index in comparison with controls.

Toxic levels of selenium in enzymes and selenium uptake in tissues of a marine fish.

Acute toxicity of selenium as selenite in Zosterisessor ophiocephalus by ip injection was studied. The 50% lethal dose and 50% lethal time were measured to be 0.29 ppm and 96 h, respectively. Se concentrations in liver, gill, skin and muscle, and Cyt. P450 level, Se-GPx, and Total GPx enzyme activities in liver were also assessed at different doses and times after injection. Starting at 0.3 ppm injected dose, enzyme activities and Se concentration in tissues, but not in muscle, showed significant differences from the control group. A threshold behavior was inferred. Normal conditions of enzyme activities and Se concentration in tissues were restored about 1 wk after injection. Biological elimination half-lives were about 2 d for liver and gill, and 5 d for skin.

Effects of copper and cadmium on growth, superoxide dismutase and catalase activities in different yeast strains.

1. Three strains of Saccharomyces cerevisiae have been adapted in vitro upon treatment with copper or cadmium. Growth rate, cellular size, metal uptake, superoxide dismutase and catalase activities were measured. 2. Growth rate and metal uptake are quite different among the yeast strains and also for copper and cadmium treatment. At the employed concentrations, only cadmium chiefly affects the cellular volume. 3. Cu, ZnSOD activity is stimulated in the presence of copper, while it is lightly inhibited in the presence of cadmium. Catalase level remains almost unchanged in the conditions tested. This lack of correlation is then discussed.

Cobalt substitution studies on bovine erythrocyte superoxide dismutase: evidence for a novel cobalt-superoxide dismutase derivative.

Three cobalt derivatives of bovine erythrocyte superoxide dismutase (superoxide:superoxide oxidoreductase, EC 1.15.1.1) have been prepared under different pH conditions using a cobalt-thiocyanate complex which has already proved to yield specific substitutions on other copper proteins. The cobalt-protein derivatives have been characterized by optical, circular dichroism and fluorescence spectroscopies. One derivative, referred to as Co2Co2-protein, contains Co(II) ions specifically bound at both Zn(II) and Cu(II) sites. On the basis of their spectroscopic properties, the other two derivatives can be referred as E2Co2- and Co2E2-superoxide dismutase, with cobalt substituting, respectively, at the zinc and the copper sites leaving the contiguous site empty (E). The Co2E2-protein complex represents a novel derivative, since it has never been described in literature. The optical spectrum in the visible region of Co2-Co2-protein well corresponds to the sum of the spectra of the other two derivatives. The circular dichroism spectrum of Co2Co2-derivative, however, is not the sum of individual E2Co2- and Co2E2-proteins, suggesting that the presence of Co(II) in one site strongly affects the geometry of the neighbouring site. Some discrepancies between our spectroscopic data and those reported in literature are discussed. The results obtained from fluorescence experiments indicate that Co(II) ions exert a different quenching effect on the tyrosine emission, depending on whether they are located in the Zn(II) or in the Cu(II) site. The fluorescence quenching can be attributed to a 'heavy atom' and 'paramagnetic ion' effect by Co(II) ions.

The role of copper and quaternary structure on the conformational properties of Octopus vulgaris hemocyanin.

Some structural properties of Octopus vulgaris hemocyanin have been investigated by fluorescence spectroscopy. The three-dimensional structure of Octopus hemocyanin is remarkably tight, resulting in a deep burial of almost all the tryptophyl residues of the protein. The hemocyanin conformation has been studied in the two main aggregation states (11 S, 50 S) of the protein, and with respect to the presence or absence of copper in the active site. Upon changing the pH of the solution, Octopus hemocyanin in the 50 S aggregation state can assume at least three different conformations. During the transition between each conformation the fluorescence quantum yield changes, but the environment of tryptophans does not change. Dissociation of the protein from 50 S to 11 S strongly enhances its susceptibility toward denaturating agents such as pH or temperature, and modifies the effects of fluorescence quenchers such as acrylamide. Moreover, these effects are more pronounced when copper is removed from the active site. A comparative analysis of the results shows that the subunit-subunit interactions exerted within the 50 S species are more important in the maintenance of the conformational stability than the copper ions present in the active sites. This behavior can be accounted for by the large amount of Ca(II) ions linked to 50 S hemocyanin.