The objective of the work was to determine the content of magnesium in plants of Trifolium pratense L. which grew in an environment contaminated with aluminium or copper. The growth and development of the plants was carried out in hydroponics, with strictly controlled composition and pH of the growth medium. Copper was added to the medium (as an additional application) in a solution of CuSO4·5H2O, and aluminium in the form of a solution of AlCl3, at concentrations of Al+3 and Cu+2 of 20 and 100 mg dm-3. The concentration of magnesium, aluminium and cooper was determined by means of a ICP sequential emission spectrometer D-820 Hilger Analytical (UK) in extracts from the roots and aboveground parts. Decreasing pH of the medium did not cause any statistically significant changes in the concentration of the metals studied. There was a significant increase in the concentration of aluminium and copper in roots and aboveground parts of Trifolium pratense L. and lower magnesium concentration for plants that were grown in an environment contaminated with Al+3/ Cu+2 at a concentration of 100 mg dm-3. In the lower concentrations of both stressors (20 mg dm-3) a decrease was noted in the level of magnesium in the whole plant, and a statistically significant increase in the concentration of aluminium and copper in the roots of Trifolium pretense L. In the aboveground parts of the plants that grew in me-dium supplemented with 20 mg dm-3 of Al+3/Cu+2, the increase of the concentration of these metals was not statistically significant.
Neurotoxic effects of copper ions after intraperitoneal (ip) and intracerebroventricular (icv) injections of CuSO4 • 5H2O and Cu(CH,COO) 2 • H2O at doses of 1-100 nmols icv and 0.1-100 pmols/kg ip was determined in rats using two behavioural methods: exploratory and locomotor activity in an open field test and spatial memory in a water maze test. It has been found that higher doses of copper salts (100 nmols icv and 100 ^mols/kg ip) significantly decreased spatial memory of rats.
We applied the whole-cell patch-clamp technique to study the inhibitory effect of copper ions (Cu) on the activity of Kv1.3 channels expressed in human lymphocytes. Application of Cu reversibly inhibited the currents to about 10% of the control value in a concentration-dependent manner with the half blocking concentration of 5.28±0.5 µM and the Hill's coefficient of 3.83±0.18. The inhibitory effect was saturated at 10 µM concentration. The inhibition was time-dependent and it was correlated in time with a significant slowing of the current activation rate. In contrast the voltage dependence of activation was not changed by Cu as well as the inativation kinetics. The inhibitory effect of Cu was voltage-independent. It was also unaffected by changing the extracellular pH in the range from 6.4 to 8.4, raising the extracellular potassium concentration to 150 mM and by changing the holding potential from -90 to -60 mV. The inhibitiory effect of Cu was not changed in the presence of an equivalent concentration of Zn. Altogether, obtained data suggest that Cu inhibits Kv1.3 channels by a different mechanism than Zn and that Cu and Zn act on different binding sites. The inhibitory effect of Cu was probably due to a specific binding of Cu on binding sites on the channels. Possible physiological significance of the Cu-induced inhibition of Kv1.3 channels is discussed.
Recent research into the structure and properties of proteins and peptides as physiologically active diet components has spurred a new interest in the isolation and investigation of bioactive peptides of animal, plant and microbiological origin. The isolation and separation of protein and peptide mixtures requires advanced procedures. It usually involves a multi-stage separation process on chromatographic columns with various packing. Immo- bilised Metal Ion Affinity Chromatography (IMAC) is frequently used in the complex process of obtaining peptide fractions. Affinity Chromatography (IMAC) relies on the specific interactions between amino acids, their reactive groups in proteins and peptides and „transitory” metal ions, in particular Cu2+. Those ions are immobilised by the chelating compound on the bed, forming specific adsorbents which bind proteins and peptides. The aim of this study was to determine whether o-phosphoserine (OPS) can be used for the immobilization of copper ions on Sephadex G25 during the separation of peptides and proteins isolated from string beans. Frozen pods of dwarf, green-podded string bean cv. Fana were used in the study. Peptide were extracted from well-homogenized string bean pods with tris-HCl buffer (pH 7.5), from which high molecular weight proteins were isolated with methanol, acetone, 20% trichloroacetic acid and the Magnafloc M-22S cation flocculant. The protein and peptide content of the separated fractions was determined. The peptide content depended on the type of extract from which high molecular weight proteins were isolated. The results obtained by using OPS as a chelating agent in the separation of string bean can be recommended for analysis of plant peptides.
The main aim of the paper was to study the effect of coagulation of copper ions on dissolved form of humic acids. Research material was isolated from four peaty muck soils (Terric Histosols). The soils were described by means of the Springer humification number, secondary transformation index, ash content and pH in H2O and in KCl. For the studied humic acids, surface charge was determined, E4/E6 which described the degree of concentration of the chain of aromatic carbons of the humic acids and could be used as their humification number, and the elemental composition and atomic ratios were determined as well. Systems composed of humic acids and increasing concentrations of copper ions were investigated at pH 5 and in the copper concentration range of 0-40 mg dm-3, at constant concentration of humic acids. Coagulation of solutions was studied by measurement of changes of organic carbon and copper ions concentration in liquid phase at pH 5 as well as by observation of absorbance changes at wavelength 465 nm in series of prepared solutions. The results showed that coagulation of experimental systems consisted mainly in humic acid coagulation. In a certain range of concentration of copper added to solutions, metal also partly precipitated and went to the solid phase together with humic carbon, which indicated coagulation of complexed compound of humic acids with copper. At high copper concentration, ionic strength had a dominant influence on coagulation of humic acids. Copper ions stayed all in liquid phase, whereas almost total organic carbon passed to precipitate, which resulted from interactions with electrolyte ions.
Sorption of copper and zinc on wheat and rye bran as well as on collagen and elastin preparations under conditions simulating human digestive system parameters was determined. Results collected during this study indicate, that sorption abilities of investigated preparations are differential and depend on the origin source, as well as pH environment. Wheat and rye bran showed higher sorption ability. The value of copper and zinc sorption was between 39-82% and was lower for collagen and elastin (15-17%). Examined preparations show higher sorption ability when pH equals 8.7 and 6.8, and lower value of this parameter in an acidic environment.
Studies on the possibilities of activating heavy metal ions by platinum ions in the soil have been undertaken. The present work gives results of research on the copper ions activated in loessial soil by the treatment with solutions containing platinum ions. Samples of podzolic soil developed from loess were collected from the horizon Ap (0-20 cm). The samples were treated with re-distilled water or with the same quantity of PtCl4 water solutions containing 2, 4, 20 mg of platinum ions. Soil reaction (pH) was measured after 1, 2, 3, 4, 5 and 6 months of incubation at 5 and 20 °C temperature. Copper ion content was determined in the filtrates using the technique of atomic absorption spectrometry. The amount of copper ions activated in the soil material depended on the soil reaction, temperature, quantity of platinum ions introduced and the time duration of the experiment.
Preincubation of rat liver nuclei with copper ions influenced the stability and protein composition of the nuclear matrices isolated by a "high salt" method. Also the specific interaction between matrix proteins and the kappa Ig matrix attachment region of DNA was affected.
Iron and copper toxicity has been presumed to involve the formation of hydroxyl radical (.OH) from H2O2 in the Fenton reaction. The aim of this study was to verify that Fe2+-O2 and Cu+-O2 chemistry is capable of generating .OH in the quasi physiological environment of Krebs-Henseleit buffer (KH), and to compare the ability of the Fe2+-O2 system and of the Fenton system (Fe2+ + H2O2) to produce .OH. The addition of Fe2+ and Cu+ (0-20 µM) to KH resulted in a concentration-dependent increase in .OH formation, as measured by the salicylate method. While Fe3+ and Cu2+ (0-20 µM) did not result in .OH formation, these ions mediated significant .OH production in the presence of a number of reducing agents. The .OH yield from the reaction mediated by Fe2+ was increased by exogenous Fe3+ and Cu2+ and was prevented by the deoxygenation of the buffer and reduced by superoxide dismutase, catalase, and desferrioxamine. Addition of 1 µM, 5 µM or 10 µM Fe2+ to a range of H2O2 concentrations (the Fenton system) resulted in a H2O2-concentration-dependent rise in .OH formation. For each Fe2+ concentration tested, the .OH yield doubled when the ratio [H2O2]:[Fe2+] was raised from zero to one. In conclusion: (i) Fe2+-O2 and Cu+-O2 chemistry is capable of promoting .OH generation in the environment of oxygenated KH, in the absence of pre-existing superoxide and/or H2O2, and possibly through a mechanism initiated by the metal autoxidation; (ii) The process is enhanced by contaminating Fe3+ and Cu2+; (iii) In the presence of reducing agents also Fe3+ and Cu2+ promote the .OH formation; (iv) Depending on the actual [H2O2]:[Fe2+] ratio, the efficiency of the Fe2+-O2 chemistry to generate .OH is greater than or, at best, equal to that of the Fe2+-driven Fenton reaction.
Although multiple factors are associated with cardiovascular pathology, there is now an impressive body of evidence that free radicals and nonradical oxidants might cause a number of cardiovascular dysfunctions. Both direct damage to cellular components and/or oxidation of extracellular biomolecules, e.g. LDL, might be involved in the aetiology of cardiovascular diseases. The key molecules in this process seem to be iron and copper ions that catalyse formation of the highly reactive hydroxyl radical. Chelation of iron ions has a beneficial effect on the processes associated with the development of atherosclerosis and formation of post-ischemic lesions. These findings are indirectly supported by the increasing body of evidence that stored body iron plays a crucial role in pathogenesis of atherosclerosis and ischemia/reperfusion injury.
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