Exchangeable Copper Research Articles

Analytical variables affecting exchangeable copper determination in blood plasma

To resolve discrepancies observed in the determination of plasma exchangeable Cu (also called direct reacting Cu or loosely bound Cu) by several methods, plasma storage techniques and various aspects of a stable isotope dilution procedure for exchangeable Cu were evaluated. Results indicated that the exchangeable Cu fraction of plasma increased with storage at room temperature, at 5 degrees C and when subjected to repeated freeze/thaw cycles. Samples could be safely stored at -65 degrees C. Exchange between added 65Cu2+ and endogenous plasma Cu rapidly went to completion in the isotope dilution procedure. Analytical results were unaffected by shaking method, sample size or the presence of heparin. A small difference was observed between serum and plasma. The determination of exchangeable Cu did not vary over a period of 4 h when plasma was exposed to 1.6 x 10(-4)-M sodium diethyldithiocarbamate (used in the isotope dilution method) but steadily increased when exposed to 1.1 x 10(-2)-M sodium diethyldithiocarbamate, which suggested that tightly bound Cu (probably in ceruloplasmin) was exchanging with isotopic tracer at the higher concentration. Determination of exchangeable Cu was constant from pH 7.2-8.5 but increased substantially at higher pH. Complete recovery of natural Cu added to plasma was obtained. Studies in solution indicated that 65Cu2+ exchanged readily with albumin- and amino acid-bound Cu. Ultrafiltration of plasma yielded a Cu fraction about half that of the exchangeable Cu fraction. We conclude that the stable isotope dilution procedure for plasma exchangeable Cu yields reliable, physiologically meaningful results.

Copper Dynamics and Impact on Microbial Communities in Soils of Variable Organic Status

The effect of soil organic status on copper impact was investigated by means of a microcosm study carried out on a vineyard soil that had been amended with varying types of organic matter during a previous long-term field experiment. Soil microcosms were contaminated at 250 mg Cu kg(-1) and incubated for 35 days. Copper distribution and dynamics were assessed in the solid matrix by a sequential extraction procedure and in the soil solution by measuring total and free exchangeable copper concentrations. Copper bioavailability was also measured with a whole-cell biosensor. Modifications of microbial communities were assessed by means of biomass-C measurements and characterization of genetic structure using ARISA (automated-ribosomal-intergenic-spacer-analysis). The results showed that copper distribution, speciation, and bioavailability are strongly different between organically amended and nonamended soils. Surprisingly, in solution, bioavailable copper correlated with total copper but not with free copper. Similarly the observed differential copper impact on micro-organisms suggested that organic matter controlled copper toxicity. Bacterial-ARISA modifications also correlated with the estimated metal bioavailability and corresponded to the enrichment of the Actinobacteria. Contrarily, biomass-C and fungal-ARISA measurements did not relate trivially to copper speciation and bioavailability, suggesting that the specific composition of the indigenous-soil communities controls its sensitivity to this metal.

Interplay between glutathione, Atx1 and copper. 1. Copper(I) glutathionate induced dimerization of Atx1

Copper is both an essential element as a catalytic cofactor and a toxic element because of its redox properties. Once in the cell, Cu(I) binds to glutathione (GSH) and various thiol-rich proteins that sequester and/or exchange copper with other intracellular components. Among them, the Cu(I) chaperone Atx1 is known to deliver Cu(I) to Ccc2, the Golgi Cu-ATPase, in yeast. However, the mechanism for Cu(I) incorporation into Atx1 has not yet been unraveled. We investigated here a possible role of GSH in Cu(I) binding to Atx1. Yeast Atx1 was expressed in Escherichia coli and purified to study its ability to bind Cu(I). We found that with an excess of GSH [at least two GSH/Cu(I)], Atx1 formed a Cu(I)-bridged dimer of high affinity for Cu(I), containing two Cu(I) and two GSH, whereas no dimer was observed in the absence of GSH. The stability constants (log beta) of the Cu(I) complexes measured at pH 6 were 15-16 and 49-50 for CuAtx1 and Cu (2) (I) (GS(-))(2)(Atx1)(2), respectively. Hence, these results suggest that in vivo the high GSH concentration favors Atx1 dimerization and that Cu (2) (I) (GS(-))(2)(Atx1)(2) is the major conformation of Atx1 in the cytosol.

Stable isotope pilot study of exchangeable copper kinetics in human blood plasma

To develop further our understanding of initial dietary copper metabolism, a method has been developed to separate plasma copper that is bound to albumin, from that bound to ceruloplasmin. This method has been tested using plasma samples from a pilot study involving six human volunteers who consumed 3 mg oral doses of the stable isotope 65Cu and gave blood samples at timed intervals up to 7 days. The results suggest that this method can be used to monitor dynamic fluctuations in newly absorbed copper over a short time frame.

On inverse carrying charges and spatial arbitrage

AbstractA dynamic trader problem under uncertain demand is presented and spatial and temporal arbitrage conditions derived from first‐order conditions. A time‐varying storage premium that explains backwardation and inverse carrying charges independent of risk‐aversion, stockouts, or convenience is developed. Random‐returns and cost‐of‐carry models are expressed as special conditions of the derived model. An applied composite‐error model is estimated using data from the London Metal Exchange's copper contract. Predictions of the model are tested against well‐known alternatives. © 2007 Wiley Periodicals, Inc. Jrl Fut Mark 27:305–336, 2007

Development of a new approach for microbial decontamination of water using modified Fenton's reaction

Microbial decontamination of water was carried out using a novel radical generating system consisting of ion exchange resin, copper and hydrogen peroxide. The system was successful in reducing the microbial load in water by more than 99% in 15 min and is effective against all the microorganisms tested. The method was also successful in decontaminating the flood water obtained from Industrial Canal and 17th Street Canal in New Orleans. Decontamination is due to the formation of hydroxyl radicals, formed during the decomposition of hydrogen peroxide by the metal-polymer complex.

Assessment of bacterial community structure in a long-term copper-polluted ex-vineyard soil

The influence of long-term copper contamination on the diversity of bacterial communities was investigated in an ex-vineyard soil. Two sites of the same area but exhibiting different 3-fold exchangeable copper (Ex-Cu) concentrations were analysed. Culturable bacterial community structure was assessed using a variety of approaches: determination of culturable bacteria number, analyses of 132 isolates, and denaturing gradient gel lectrophoresis (DGGE) patterns of bacterial biomass grown on agar plates and of soil DNA. There was no significant difference in the number of total heterotrophs at the two sites, whereas the percentage of fast-growing bacteria growing in 1 day, was lower at the site with the higher Ex-Cu content. A high percentage of Cu-tolerant bacteria was found in both sites (63-70%) and it was relatively independent of the Cu content. Shifts in species composition of the culturable bacterial community were detected by analysing isolates from the two soils, Gram-positive bacteria prevailed in the less-polluted soil while Gram-negative bacteria in the more-polluted soil. Each sample site had a community with a different metal resistance pattern. Our study seems to indicate that in this soil ecosystem, copper influenced the culturable bacterial communities, affecting the structural diversity and altering some of the metal resistance of the microorganisms. The Sorensen similarity index calculated on DGGE profiles of 16S rDNA of total and culturable bacterial communities indicated a different species composition at the two sites, although both sites had the same biodiversity degree and different dominance.

An empirical analysis of commodity pricing

AbstractCommodity pricing models generally explain the link between commodity prices and stock levels in terms of a stock‐out constraint or a convenience yield. Analysis of this link is provided using monthly London Metals Exchange copper, lead, and zinc prices obtained for the period November 1964 to December 2003. A Markov model, fitted to these data, supports the existence of two distinct pricing regimes while the impact of convenience yields is also identified. © 2006 Wiley Periodicals, Inc. Jrl Fut Mark 26:391–415, 2006

Effects of some drugs on purified human erythrocyte CuZnSOD and in vitro inhibitiory effect of 5-fluorouracil on leukocyte total SOD activity

The inhibition and activation effects of some drugs on the activities of superoxide dismutase enzymes (SOD) in human erythrocyte and leukocyte cells was investigated. Firstly, CuZnSOD enzyme was purified 837–fold and 12% efficiency from human erythrocytes by ethanol-chloroform treatment to remove hemoglobin and then ion exchange chromatography (DEAE-Sepharose) and copper chelate affinity chromatography techniques. Inhibition or activation effects of fourteen drugs on CuZnSOD was investigated. None of the studied drugs except for 5-fluorouracil showed any effects on the enzyme. 5-fluorouracil showed activation effects on CuZnSOD at 3.33 mg/ml and 4 mg/ml concentrations with 33% and 32% activation, respectively. Leukocytes were isolated from healthy human blood, lysed in liquid nitrogen and the effect of 5-fluorouracil on the lysate SOD activity investigated. 5-Fluorouracil showed inhibition effects on total SOD activity of human leukocytes at 2 mg/ml and 4 mg/ml concentrations with 42% and 62% inhibition, respectively.

Significance of physicochemical forms of storage in microalgae in predicting copper transfer to filter-feeding oysters (Crassostrea gigas)

Copper distribution has been examined in two microalgae (Haslea ostrearia, Diatom; Tetraselmis suecica, Prasinophyceae) exposed to Cu at 30 microg/L(-1). Exchangeable copper linked at the cell surface was desorbed using 8-hydroxyquinoline-5-sulfonate as complexing agent. Then, incorporated copper was separated between soluble and insoluble fractions. In addition, algae were resuspended in acid solutions, the pHs of which covered the range existing in the digestive tract of bivalves. Considering that the soluble fraction is the most easily transferred in the food chain and that exchangeable Cu is easily desorbed, the percentages of Cu potentially available in microalgae have been assessed. These percentages have been compared with those retained in oysters Crassostrea gigas fed with contaminated microalgae in previous studies. In H. ostrearia, the potentially available fraction of Cu (90%) was very similar to the percentage retained by oysters (93%) when the bivalves were acclimated to this food for 3 weeks. Only half (21%) of the potentially available Cu of T. suecica (42%) was readily assimilated in oysters after 3 weeks. This is in agreement with the results of the desorption tests at physiological pHs which showed that only 15-25% of Cu was lost, despite solubilization of other constituents of T. suecica as demonstrated by the decrease in their dry weight. Bioavailability determined from metal speciation in food allows a relevant prediction of the trophic transfer in the case of H. ostrearia, but caution is recommended in generalizing this mode of assessment as shown in the case of T. suecica.

The storage of sewage sludge: Influence of liming on the evolution of copper chemical fractions

Sewage sludge re-used in agriculture has to be stabilized and is often stored for several months before land spreading. Stabilization treatment may affect the behaviour of heavy metals such as Cu, which is an element potentially toxic to the environment. In the present study, the chemical forms of copper have been investigated in heaps of limed and unlimed sludge coming from the wastewater treatment plant of Roselies (Belgium). These limed and unlimed aerobically digested sludges were stored during 4 mo in controlled conditions close to field ones. The sequential extraction procedure developed by Tessier was used to determine the copper chemical forms of representative samples taken in the outer shell and in the depth of the heaps. The physico-chemical properties (pH, organic matter, dry matter and temperature) of these samples were also monitored. This study shows that liming transforms part of organically-bound copper into both exchangeable and residual copper. These changes mainly occur during the first 2 wk of storage. After several months of storage, copper passes from the residual fraction to the exchangeable and oxide fractions. During the whole experiment, changes occur faster in the outer shell than in the depth of the heaps. Thus, we demonstrate that the distribution of the copper forms depends on the storage time and it is different in the outer shell from that in the depth of the heaps.

Detection of micro-flaws on thin copper tubes using SQUID-NDI system based on eddy current technique

We constructed a SQUID-NDI system for inspection of micro-flaws on heat exchanger copper tubes employing HTS-SQUID gradiometer and Helmholtz-coil-type inducer. The HTS-SQUID gradiometer was cooled using a coaxial pulse tube cryocooler. The detection of artificial flaws several tens of μm in depth on copper tubes, 6.35mm in outer diameter and 0.825mm in thickness, was demonstrated using the SQUID-NDI system. With excitation field of 1.6μT at 5kHz, the system could detect a 30-μm-depth flaw with SN ratio of about 20, where conventional NDI methods should fail to detect. The amplitude of the magnetic signal due to the flaw was in proportion to the square of flaw depth. Taking into account of the system’s noise level, the results suggest that the SQUID-NDI system has a potential to detect sub-10-μm-depth flaws on copper tubes.

High variability of the metal content of tree growth rings as measured by synchrotron micro x‐ray fluorescence spectrometry

AbstractSynchrotron radiation analysis was used to investigate the metal content of tree rings collected from paper birch, Betula papyrifera Marsh, on transects downwind from two metal smelters (nickel and copper). Individual trees reflected changes in ring metal content with time, which may be presumed to represent changes in local metal bioavailability. However, between‐tree variations were large and no statistically significant differences in metal content as a function of time were found within or between sites. Although concentrations of both total and exchangeable copper and nickel in the soil increased with proximity to the respective smelter, this pattern was reflected only in the nickel content of rings near the nickel smelter; copper content did not vary with distance from either smelter. The sites did differ with respect to lead, manganese and zinc content of the rings, which may be related to pH. In conclusion, the variability between trees at each site suggests that dendroanalysis is a poor method for evaluating metal exposure at a large (site) scale. Tree ring metal content may be used to evaluate the metal uptake by individual trees but metal mobility in the stem makes it difficult to establish a reliable chronology. Copyright © 2005 John Wiley & Sons, Ltd.

Removal of Copper from Contaminated Soil by Use of Poly(amidoamine) Dendrimers

This study characterizes poly(amidoamine) (PAMAM) dendrimers of various generations and terminal functional groups for removal of copper(II) in a sandy soil. Effects of dendrimer dose, generation number, pH, terminal functional groups, and ionic strength on the removal efficiency were investigated through a series of column tests. Over 90% of copper initially sorbed in the soil was removed by use of approximately 66 bed volumes of 0.10% (w/w) of a generation 4.5 dendrimer with carboxylate terminal groups at pH 6.0. On the basis of equal equivalent dose, dendrimers of lower generation removed more copper. Lowering pH enhanced copper removal for all dendrimers tested. In contrast, types of terminal groups (carboxylate, amine, or hydroxyl) showed a modest effect on the removal efficiency. Results from a sequential extraction procedure suggested that dendrimers removed primarily exchangeable and carbonate-bound copper. The residual copper in treated soil is predominantly bound with soil organic matter (SOM), which is much less available physical-chemically or biologically. Spent dendrimers were recovered through nanofiltration with a commercially available nanofilter. Upon acid regeneration, recovered dendrimers were reused and performed as well as the virgin dendrimers. The dendrimers may be used as reusable, high-capacity extracting agents for in situ removal of heavy metals from contaminated soils.

Dialysis-Chelex method for determination of exchangeable copper in human plasma.

A method based on dialysis in the presence of histidine, and subsequent copper adsorption and preconcentration using Chelex-100 resin, has been developed for selective extraction of exchangeable copper from blood plasma. The method was verified by comparative analysis using two different instrumental techniques, ICP-MS and AAS. The results obtained were in excellent agreement. Accurate quantification of the exchangeable copper and the firmly bound copper was achieved. The method was further validated by analysis of a lyophilised human serum certified reference material.

Speciation of copper and zinc in natural freshwater: comparison of voltammetric measurements, diffusive gradients in thin films (DGT) and chemical equilibrium models

In situ measurements of copper and zinc using diffusive gradients in thin films (DGT) in two distinct natural water systems were compared to metal speciation assessed by competitive ligand exchange (CLE) and voltammetric measurements. In a dynamic river system, where dissolved metal concentrations vary significantly over short-time periods, DGT technique provided averaged values of the metal concentrations over time. In microcosms, at different total dissolved concentrations of copper and zinc, DGT technique measured a similar fraction as measurements of labile metal performed by voltammetry. The proportion of DGT and voltammetric-labile zinc to dissolved zinc was 61±4% and, respectively, 76±9%. DGT technique was measuring 81±8% of exchangeable copper (by exchange with catechol). These two fractions were similarly influenced by the addition of NTA. In the absence of NTA, copper measured by DGT represented 34±4% of dissolved copper whereas in the presence of NTA, this proportion raised to 57±2%. These measurements were compared to calculations performed with speciation programs using several models for the complexation by humic and fulvic substances, namely Model VI (WHAM), NICA-Donnan and SHM. The predicted speciation by these three models was similar. The prediction of free zinc ion and labile zinc concentrations were in agreement with experimental data. Calculated concentrations of free copper ion were overestimated because these models are not considering strong specific copper-binding ligands probably present in natural water.

Desulfurization of diesel fuels by adsorption via pi-complexation with vapor-phase exchanged Cu(I)-Y zeolites.

Desulfurization of a commercial diesel fuel by vapor-phase ion exchange (VPIE) copper(I) faujasite zeolites was studied in a fixed-bed adsorber operated at ambient temperature and pressure. The zeolite adsorbed approximately five thiophenic molecules per unit cell. After treating 18 cm3 of fuel, the cumulative average sulfur concentration detected was 0.032 ppmw-S. GC-FPD results showed that the pi-complexation sorbents selectively adsorbed highly substituted thiophenes, benzothiophenes, and dibenzothiophenes from diesel, which is not possible by using conventional hydrodesulfurization (HDS) reactors. The high sulfur selectivity and high sulfur capacity of the VPIE Cu(II)-zeolites were due to pi-complexation.

Evaluation of factors influencing root-induced changes of copper fractionation in rhizosphere of a calcareous soil

Major factors influencing the root-induced copper fractionation changes within the rhizosphere of maize, wheat, pea, and soybean seedlings were evaluated using a contaminated calcareous soil. The effects of acidification, alkalization, and introduction of root exudates were investigated by addition of acid, alkaline and root exudates from solution cultures, prior to incubation and copper fractionation. Raw and sterilized soils were compared for changes of copper fractionation in the rhizosphere using rhizoboxes with maize, wheat, pea and soybean seedlings. The results indicated that the general trend in considerable changes was similar among the plant species studied. The rhizosphere experienced a depletion of carbonate associated and organic bound copper along with an accumulation of exchangeable and Fe–Mn oxide bound copper. The resulting significant influence of root exudates on copper fractionation appears to have been produced through complexation rather than acidification or alkalization. The increase in exchangeable copper in rhizosphere was strengthened by microorganisms.

Accumulation of copper and zinc in periphyton in response to dynamic variations of metal speciation in freshwater.

Although the free ion activity model (FIAM) has been well-established in laboratory studies, there remains the need for field data in order to validate the applicability of this model in natural systems. The objective of this study was to investigate the response of copper and zinc accumulation in periphyton to short-term variations in metal concentration and speciation in freshwater. During heavy rain events, dissolved Cu in the Furtbach stream increased from 40 to 118 nM, while dissolved Zn increased from 45 to 147 nM due to the release of metals from contaminated sediments. Increases in free copper and free zinc ions in the water (from 10(-14) to 10(-11.5) M for Cu2+; from 1 to 15 nM for Zn2+) were observed during the onset of heavy rain events. Periphytic algae collected from artificial substrates had an intracellular copper content (0.2-2.8 micromol/g dry weight (dw)) that varied as a function of the exchangeable copper in the water (labile form) rather than the free Cu2+. Intracellular zinc content (1.5-8.0 micromol/g dw) was found to follow the same trend as the free zinc ion concentration. Adsorbed Cu and Zn on periphyton showed a very dynamic response to variations in dissolved metal concentration. Different concentrations of dissolved manganese during the two time periods may affect the accumulation of zinc and copper by competition for metal uptake.

Interactions of Copper with Glycated Proteins: Possible Involvement in the Etiology of Diabetic Neuropathy

Humans and animals with diabetes frequently develop peripheral vascular dysfunction and peripheral neuropathies. There is accumulating evidence that impaired peripheral nerve function may derive from diminished endoneural blood flow. The decrements in nerve blood flow may, in turn, be due to diminished endothelium‐dependent vasodilation. Although a number of possible causes of this defective vasodilation have been suggested, none has been definitely proven. Regardless of the precise cause, the impaired vasodilatory activity may reflect diminished availability of endothelium‐derived relaxing factor (EDRF), variously thought to be nitric oxide or thiol adducts of nitric oxide. Other investigators have reported that administration of transition metal chelators to diabetic rats corrects EDRF‐mediated arterial relaxation and restores both neural blood flow and nerve conduction velocity, suggesting the involvement of transition metals. Our investigations center about the hypothesis that glycated proteins bind transition metals such as copper and iron, and that such ‘glycochelates’ accumulate within the vasculature in diabetes and catalytically inactivate EDRF. In partial support of this hypothesis: (1) Glycated albumin binds 3‐fold greater amounts of both copper and iron. (2) Copper bound to glycated albumin remains redox active (e.g. capable of supporting the oxidation of ascorbic acid). (3) Copper and copper‐containing glycochelates cause the rapid decomposition of one putative form of EDRF, nitrosocysteine. (4) The amount of exchangeable (i.e. chelatable) copper in the plasma of diabetic rats is approximately twice that in normal rat plasma. (5) Similarly, tail tendons of diabetic animals have about twice as much bound copper as do tendons of normal rats. (6) Implants bearing adsorbed glycated albumin placed in the peritonea of normal mice for 48 h accumulate 5 times as much bound copper as do implants coated with control albumin. Overall, these observations support—but do not conclusively prove—the hypothesis that transition metals such as copper, bound to glycated proteins, may blunt normal EDRF‐dependent relaxation of diabetic arteries and provide a rationale for the use of transition metal chelators in the therapy of diabetic vasculopathy and neuropathy.