Quantitative investigation of copper(II) and zinc(II) complexes with S-carboxymethyl-L-cysteine and computer-simulated appraisal of their potential significance in vivo

S-carboxymethyl-L-cysteine (SCC) is a mucolytic agent extensively used in the treatment of respiratory tract disorders. Some of the undesirable side effects observed during SCC therapy being reminiscent of symptoms characteristic of copper and zinc imbalances, the objective of this paper was to test the possible interference of SCC with the metabolism of these two metals. Copper(II)- and zinc(II)-SCC complex equilibria have thus been investigated under physiological conditions by means of classical potentiometry combined with computer-assisted calculation techniques. Formation constants derived from these studies have then been used to simulate 1) the potential influence of SCC on the distribution of the above metals in blood plasma and 2) the extent to which gastrointestinal interactions between the drug and each metal ion in turn are likely to affect the bioavailability of each other. The results of these simulations show that 1) plasma therapeutic levels of SCC are not likely to induce dramatic changes in the distributions of copper(II) and zinc(II) low molecular weight fractions, 2) the gastrointestinal distribution of the drug is not affected by standard dietary doses of these metals, and 3) in contrast, therapeutic concentrations of SCC are capable of mobilizing significant fractions of both metals into tissue-diffusible electrically neutral complexes. In conclusion significant depletions of neither copper nor zinc are to be expected from oral administration of SCC. While the drug may to some extent facilitate the excretion of Cu2+ and Zn2+ ions from blood plasma, its gastrointestinal influence is, on the contrary, favorable to a better absorption of these two metals.     

Aluminum speciation studies in biological fluids. Part 7. A quantitative investigation of aluminum(III)-malate complex equilibria and their potential implications for aluminum metabolism and toxicity.

As a nonessential element, aluminum may be toxic at both environmental and therapeutic levels, depending on ligand interactions. Dietary acids that normally occur in fruits and vegetables and commonly serve as taste enhancers are good ligands of the Al(3+) ion. Malic acid is one of these and also one of the most predominant in food and beverages. The present paper reports an examination of its potential influence on aluminum bioavailability through speciation calculations based on Al(III)-malate complex formation constants especially determined for physiological conditions. According to the results obtained, malate appears to be extremely effective in maintaining Al(OH)(3) soluble over the whole pH range of the small intestine under normal dietary conditions. In addition, two neutral Al(III)--malate complexes are formed whose percentages are maximum from very low malate levels. When aluminum is administered therapeutically as its trihydroxide, the amount of metal neutralized by malate peaks as its solubility pH range regresses to its original limits in the absence of malate. The enhancing effect of malate towards aluminum absorption is therefore virtually independent of the aluminum level in the gastrointestinal tract. The presence of phosphate in the gastrointestinal juice is expected to limit the potential influence of malate on aluminum absorption. Under normal dietary conditions, phosphate effectively reduces the fraction of aluminum neutralized by malate but without nullifying it. Aluminum phosphate is predicted to precipitate when aluminum levels are raised as with the administration of aluminum hydroxide, but a significant amount of neutral aluminum malate still remains in solution. Even therapeutic aluminum phosphate is not totally safe in the presence of malate, even at low malate concentrations. As plasma simulations predict that no compensatory effect in favor of aluminum excretion may be expected from malate, simultaneous ingestion of malic acid with any therapeutic aluminum salt should preferably be avoided.     

Metal ion-tetracycline interactions in biological fluids. 2. Potentiometric study of magnesium complexes with tetracycline, oxytetracycline, doxycycline, and minocycline, and discussion of their possible influence on the bioavailability of these antibiotics in blood plasma

The formation constants of the various complexes formed by magnesium with four tetracycline derivatives, namely, tetracycline itself, oxytetracycline, doxycycline, and minocycline, were determined by potentiometry over large pH ranges under experimental conditions pertaining to blood plasma (37 degrees C, NaCl 0.15 mol dm-3). The results were used, together with those previously obtained on the complexation of these tetracyclines with proton and calcium, to assess the influence of the two alkali earth metal ions on the bioavailability of these drugs in blood plasma. Accordingly, simulations of the distribution of the four tetracyclines into their different proton and metal complex species were calculated. The distributions confirm that, in combination with the protein-bound fraction of the tetracyclines, the metal-bound fraction represents more than 99% of these drugs in plasma, the extent of their free fraction commonly being less than 1%.     

Copper‐resistant halophilic bacterium isolated from the polluted Maruit Lake,Egypt

Aims: To isolate and characterize copper‐resistant halophilic bacteria from the polluted Maruit Lake, Egypt and identify the role of plasmids in toxic metal resistance. Methods and Results: We isolated strain MA2, showing high copper resistance up to the 1·5 mmol l^?1 concentration; it was also resistant to other metals such as nickel, cobalt and zinc and a group of antibiotics. Partial 16S rRNA analysis revealed that strain MA2 belonged to the genus Halomonas. Copper uptake, measured by atomic absorption spectrophotometery, was higher in the absence of NaCl than in the presence of 0·5–1·0 mol l^?1 NaCl during 5–15 min of incubation. Cell fractionation and electron microscopic observation clarified that most of the copper accumulated in the outer membrane and periplasmic fractions of the cells. Plasmid screening yielded two plasmids: pMA21 (11 kb) and pMA22 (5 kb). Plasmid curing resulted in a strain that lost both the plasmids and was sensitive to cobalt and chromate but not copper, nickel and zinc. This cured strain also showed weak growth in the presence of 0·5–1·0 mol l^?1 NaCl. Partial sequencing of both plasmids led to the identification of different toxic metals transporters but copper transporters were not identified. Conclusions: The highest cell viability was found in the presence of 1·0 mol l^?1 NaCl at different copper concentrations, and copper uptake was optimal in the absence of NaCl. Plasmid pMA21 encoded chromate, cobalt, zinc and cadmium transporters, whereas pMA22 encoded specific zinc and RND (resistance, nodulation, cell division) efflux transporters as well as different kinds of metabolic enzymes. Copper resistance was mainly incorporated in the chromosome. Significance and Impact of the Study: Strain MA2 is a fast and efficient tool for copper bioremediation and the isolated plasmids show significant characteristics of both toxic metal and antibiotic resistance.     

Influence of dietary carbohydrate on copper, iron, and zinc status of the rat.

Rats were fed diets containing starch, sucrose, glucose, or fructose as the carbohydrate source, and the influence of these carbohydrates on copper, iron, and zinc status was determined. It was found that copper absorption was reduced in animals receiving glucose. This reduction was exaggerated when a high level of iron was present in the diet, indicating a possible antagonism between iron and copper at the site of absorption. Iron and zinc status of the animals also appeared to be influenced to some extent by dietary carbohydrate.     

Trace elements in diabetes mellitus. Peculiarities and clinical validity of determinations in blood cells

Significantly more information about trace element status can be obtained by investigating concentrations in blood cells instead of only evaluating the concentrations in plasma. This can be explained by the fact that essential trace elements such as zinc, copper, chromium and selenium take part in a variety of enzymatic processes on a molecular cellular level. Ignoring these important biochemical roles, trace element concentrations determined in whole blood or plasma very often lead to conclusions contrary to the actual intracellular concentration. Especially in metabolic diseases like diabetes mellitus, conclusions drawn from trace element concentrations in blood cells usually offer more valuable clinical information about the metabolic state than trace element concentrations in plasma or whole blood. In the present investigation copper and zinc concentrations were increased in all blood fractions of diabetic patients (IDDM). In insulin-dependent diabetic children significantly higher values of zinc in erythrocytes were also found, and they were higher in patients with poor metabolic control (HbA1c>9%). When different blood fractions in diabetic patients (NIDDM) were compared with a control group, chromium was significantly increased in plasma and polymorphonuclear cells. Patients with IDDM had pronounced decreased selenium concentrations in erythrocytes as compared to controls.     

Patients at risk for trace element deficiencies: Bariatric surgery

Obesity is a worldwide epidemic associated with diseases such as diabetes mellitus and cardiovascular disease. Current methods for weight loss are not very effective, particularly for those with morbid obesity. Surgical therapy may be recommended for those with a BMI ≥ 40 kg/m², or BMI ≥ 35 kg/m² with co-morbidities. This therapy can produce significant weight loss and improve/resolve co-morbidities including hypertension and hyperlipidemia. Yet successes may be tempered by adverse effects on trace element absorption and status. A PubMed literature search identified studies from January 1980 to February 2013 for inclusion in a meta-analysis. Publications that contained keywords ‘bariatric surgery or gastric bypass,’ ‘trace element or mineral or zinc or iron or copper or iodine or manganese’, and ‘absorption or status or rate or level’ were identified. Inclusion criteria were human markers that reflect changes in trace element status before and after bariatric surgery. The meta-analysis found a decrease in blood copper, zinc, hemoglobin, as well as an increase in iron, regardless of the type of surgery. The pooled effect sizes and 95% confidence intervals were 0.17 and −0.09 to 0.43 for plasma/serum iron (p = 0.20); −0.49 and −0.67 to −0.31 for blood hemoglobin (p = 0.00); −0.47 and −0.90 to −0.05 for plasma/serum copper (p = 0.03); −0.77 and −1.20 to −0.35 for plasma/serum zinc (p = 0.00). Differences in levels of these minerals pre- and post-surgery may have been influenced by the time period after surgery, a pre-existing deficiency, type and dose of vitamin–mineral supplements, and malabsorption due to elimination of parts of the gastrointestinal tract.     

The treatment of Wilson's disease with zinc. IV. Efficacy monitoring using urine and plasma copper

Progress has been made in establishing the efficacy and safety of oral zinc as a maintenance therapy for Wilson's disease. It is important to develop simple, noninvasive monitoring methods to assure the adequacy of zinc therapy in individual patients. In this paper we report the use of 24-hr urine copper and plasma copper measurements to monitor efficacy of zinc maintenance therapy in 30 Wilson's disease patients. In examples of therapeutic inadequacy such as noncompliance, these values increase. With continued long-term adequate therapy, they remain stable or decrease. These two simple monitoring tools appear to be very useful in monitoring Wilson's disease patients receiving zinc therapy.     

Inhibition of copper absorption by zinc

Copper and zinc interact at the intestinal mucosal level, affecting copper absorption. Amino acids, such as histidine, may affect the absorption of these two elements by chelating these cations. The two mechanisms could have additive potential. This possibility was investigated using a duodenal-jejunal single-pass perfusion procedure in anesthetized rats. Copper absorption and tissue retention from solutions containing 0.1 mM copper were determined in the presence of either no zinc or equimolar zinc, or at a zinc/copper ratio of 10/1, either without histidine or with histidine at a 10/1 or 20/1 ratio to copper. Copper removal from the intestinal lumen was decreased by zinc, and further reduced by increasing concentrations of histidine. There was a greater accumulation of copper in the small intestine, reaching a maximum with a 10-fold excess of histidine. With zinc at a 10/1 ratio to copper, the addition of a 10- or 20-fold molar excess of histidine further decreased the net uptake of copper from the perfusate while greater copper accumulation in the tissue occurred. Histidine thus enhances the inhibitory effects of zinc on copper absorp|tion, suggesting the application of convergent mechanisms for diminishing copper uptake. This could be relevant for the treatment of Wilson’s disease.     

Trace metal requirements in total parenteral nutrition. Part 6. A quantitative study of the copper(II)histidine ternary complexes with leucine,glutamic acid,methionine, tryptophan and alanine,and final evaluation of the daily doses of copper and zinc specific to a nutritive mixture of a given composition

A computer-based approach was previously developed which made it possible to interpret the origin of the trace metal extra-losses induced by total parenteral nutrition (TPN) on a quantitative basis. The TPN-induced abnormal excretions of zinc and copper have been attributed to the enhanced mobilization of these metal ions into their plasma low molecular weight fraction by specific components of the nutritive solution, the mode of excretion (urinary for zinc/biliary for copper) being dictated by the electrical charge of the prevailing complexes. This implies that the trace metal needs during TPN must depend on the composition of the infusate.It was thus proposed that every nutritive solution should contain total trace metal in such an amount that the corresponding free concentration be the same as that pertaining to normal blood plasma. In this way, the metal losses would not occur at the expense of the patient's body reserve.Nevertheless, two conditions must be fulfilled before reliable doses can be calculated for a given nutritive mixture: (i) precise free metal ion concentrations in normal blood plasma must be assessed, (ii) a computer-based reliable distribution of each metal ion in the nutritive solution must be obtained. Now, the reliability of the involved simulation model directly depends on that of the equilibrium constants of the predominant complexes. A series of determinations was thus successively carried out for zinc and copper until a realistic degree of reliability (over 80%) was reached.The present paper focuses on the determination of the formation constants of the mixed-ligand complexes formed by copper and histidine with five amino acids prevailing in the nutritive mixture under study, which brings the degree of reliability of the copper distribution up to 84%.The final estimation of the zinc and copper absolute doses for the nutritive solution chosen as a reference is discussed with regard to the limitations of the present approach for which a number of applications are suggested.     

Copper metabolism in analbuminaemic rats fed a high-copper diet

Copper metabolism in male Nagase analbuminaemic (NA) rats was compared with that in male Sprague Dawley (SD) rats fed purified diets containing either 5 or 100 mg Cu/kg diet. Dietary copper loading increased hepatic and kidney copper concentrations in both strains to the same extent, but baseline values were higher in the NA rats. There was no strain difference in true and apparent copper absorption nor in faecal endogenous and urinary copper excretion. NA rats had higher levels of radioactivity in kidneys at 2 hr after intraperitoneal administration of ⁶⁴Cu. As based on the distribution of added ⁶⁴Cu, about 70% of plasma copper appeared to be in the non-protein compartment in the NA rats, whereas in SD rats, it was only about 1%. It is concluded that the NA rats are able to maintain a relatively normal metabolism of copper, even after dietary copper challenge. In the NA rats, zinc concentrations in kidneys, liver and urinary zinc excretion were elevated when compared with SD rats. The high-copper diet did not affect tissue zinc concentrations and apparent zinc absorption in both strains of rats.     

Correlation between maternal plasma homocysteine and zinc levels in preeclamptic women

Women with preeclampsia have been shown to have elevated blood levels of the metabolite homocysteine, and alterations in blood levels of zinc and copper have also been reported. This study measured plasma levels of zinc, copper, and homocysteine in women with preeclampsia and in women with healthy, normotensive pregnancies. For the patients with preeclampsia compared with controls, significantly higher mean plasma levels were found of homocysteine (16.39 vs 9.45 nmol/mL; p≤0.001), zinc (15.53 vs 11.93 μg/g protein; p < 0.05), and copper (47.90 vs 31.60 μg/g protein; p=0.001). The ratio of plasma Cu/Zn levels tended to be higher in preeclamptic women and could be taken as an index of inflammatory reaction, but the difference was not significant. Homocysteine concentrations correlated positively with plasma zinc concentrations in women with preeclampsia (r=0.588, p=0.003) but not in women with healthy pregnancies. No correlations were observed between plasma levels of homocysteine and copper. Thus, the present study found evidence that preeclampsia might be associated with hyperhomocysteinemia and elevated blood levels of zinc and copper. Furthermore, elevated blood levels of zinc were significantly associated with hyperhomocysteinemia in preeclampsia. More studies are warranted to investigate further any relationship between altered homocysteine metabolism and levels of zinc and copper in preeclampsia.     

Effects of interaction between65Zn,cadmium, and copper in rats

Distribution and retention of zinc in the presence of cadmium and copper was studied in rats exposed repeatedly to these metals. The experiment was performed on white rats of the Wistar strain. The animals were divided into four groups/five rats each: 1)⁶⁵ZnCl₂; 2)⁶⁵ZnCl₂+CdCl₂; 3)⁶⁵ZnCl₂+CuCl₂; and 4) control group. Rats were administered sc every other day for two weeks:⁶⁵ZnCl₂−5 mg Zn/kg; CdCl₂−0,3 Cd/kg; and CuCl₂−2 mg Cu/kg. The zinc content was measured in rat tissues by γ-counting. Effect of Cd and Cu on subcellular distribution of zinc in the kidney and liver and on the level of metallothionein were also examined. Whole body retention of zinc under the influence of cadmium was lower than that observed in animals treated with zinc alone. However, copper increased twofold the whole body retention of zinc. Cadmium elevated the accumulation of zinc only in the kidneys nuclear fraction and liver soluble fraction. In the kidneys and liver, copper elevated the accumulation of zinc, in the nuclear, mitochondrial, and soluble fractions. The level of metallothionein-like proteins (MT) in the kidneys after a combined supply of zinc and copper was significantly increased with respect to the group of animals treated with zinc alone. These results indicated complex interactions between cadmium, copper, and zinc that can affect the metabolism of each of the metals.     

Rapid and direct determination of selenium, copper, and zinc in blood plasma by flow injection-inductively coupled plasma-mass spectrometry

A flow injection-inductively coupled plasma-mass spectrometry (FI-ICP-MS) with a simple sample preparation procedure was developed for the determination of selenium, copper, and zinc in blood serum/plasma. A serum/plasma sample was filtered through a 0.45-μm membrane filter and diluted with a mixture of trace elements in a standard solution (9∶1, v/v). Measurement of the reference serum sample confirmed the accuracy of our method for selenium, copper, and zinc concentration. In the case of blood plasma samples obtained from six healthy adult males, the selenium, copper, and zinc concentrations were similar to those of a typical healthy male in Japan. These results suggest that the sample prepartive procedure coupled with FI-ICP-MS can be used for the routine determination of selenium, copper, and zinc in human blood serum/plasma.     

Effect of end-stage renal disease and diabetes on zinc and copper status

The aim of this study was to compare the nutritional status of zinc and copper in patients with and without diabetes submitted to chronic hemodialysis. Thirty-three patients with type 2 diabetes (DM group), 30 nondiabetic patients (NDM group), and 20 healthy individuals (control group) were studied. Plasma, erythrocyte, and urinary zinc and plasma copper were obtained from atomic absorption spectrophotometry and ceruloplasmin by immunonephelometry. The anthropometric parameters were similar among the groups. Plasma zinc was lower and erythrocyte zinc was higher in the DM and NDM groups in relation to the control group. No difference in urinary zinc was observed comparing the groups. Plasma copper was higher in the DM group when compared to the NDM and control groups. Ceruloplasmin was similar in the three groups. Serum urea was a positive independent determinant of plasma zinc concentrations. The determinants of erythrocyte zinc were MAMC midarm nuscle circumference and Kt/V dialysis adequacy. The determinants of plasma copper concentration were serum creatinine and serum glucose. The results of this study demonstrate an alteration in the distribution of zinc of patients with chronic kidney disease (CKD) independently of the presence of DM. Also, the status of copper seems not to be influenced by CKD, but only by the metabolic derangements associated with diabetes.     

Electrolyte composition of cow and calf tissues in health and in acute digestive disturbances]

Sodium, potassium, calcium, magnesium, manganese, zinc and chloride contents in the blood plasma of calves at parturition have been established to correspond to their levels in blood of cows during their calving period. Iron and inorganic phosphorus contents in calves blood plasma appeared to increase and copper content to be lower in this period as compared to postnatal period. By the third day of postnatal ontogenesis sodium concentration in calves blood decreased, copper level increased and the rest indices of water-salts metabolism in calves and adult animals were alike. Digestion disturbances in the calves were accompanied by changes in levels and magnitude of Na+/K+ ratio, magnesium, iron, copper, manganese and zinc contents in blood, liver and kidneys as well as Ca++/Pi ratio in mitochondria and cytosol of liver and jejunum mucose layer cells in comparison with clinically healthy animals.     

Aluminum speciation studies in biological fluids. Part 6. Quantitative investigation of aluminum(III)-tartrate complex equilibria and their potential implications for aluminum metabolism and toxicity

Recent epidemiological studies have confirmed the existence of a correlation between aluminum level in low-silica drinking water and prevalence of Alzheimer's disease. Also, oral aluminum-based phosphate binders and antacids may induce acute aluminum toxicity. Whatever the source of the metal ingested, its bioavailability is a function of the chemical forms under which it occurs in the gastrointestinal tract, i.e. of the ligands with which the Al3+ ion may associate. Dietary acids in particular can favor the bioavailability of aluminum in different ways: by increasing its solubility, by complexing it into neutral species, and/or by acting indirectly on its absorption process. Among these, tartaric acid is commonly found in fruits and in industrial foods and drinks, and may therefore be ingested together with environmental or/and therapeutic aluminum. The present work examines its potential influence on aluminum bioavailability. Firstly, Al(III)-tartrate complex formation constants have been determined under physiological conditions (37 degrees C, 0.15 M NaCl). Then these constants have been used to simulate the influence of tartrate on aluminum speciation in different gastrointestinal situations in which phosphate was also taken into account. Under normal conditions of aluminum contamination, tartrate is expected to keep the metal soluble throughout the whole pH range of the small intestine, which is likely to enhance its bioavailability. Even at low concentrations, tartrate also gives rise to two neutral complexes that span over the 1.5-7.5 pH interval, a phenomenon that is aggravated by increased aluminum levels as may result from aluminum hydroxide therapy. The co-occurrence of dietary phosphate reduces the fraction of aluminum neutralized by tartrate under normal conditions, but this effect quickly decreases with increasing aluminum doses. Even the therapeutic use of aluminum phosphate is not expected to be totally safe in the presence of tartaric acid. As plasma simulations show that no aluminum mobilization can be expected from tartrate that could enhance aluminum excretion, avoiding ingestion of tartaric acid during any form of aluminum-based therapy appears advisable.     

Menkes copper-translocating P-type ATPase (ATP7A): biochemical and cell biology properties,and role in Menkes disease

The Menkes copper-translocating P-type ATPase (ATP7A; MNK) is a ubiquitous protein that regulates the absorption of copper in the gastrointestinal tract. Inside cells the protein has a dual function: it delivers copper to cuproenzymes in the Golgi compartment and effluxes excess copper. The latter property is achieved through copper-dependent vesicular trafficking of the Menkes protein to the plasma membrane of the cell. The trafficking mechanism and catalytic activity combine to facilitate absorption and intercellular transport of copper. The mechanism of catalysis and copper-dependent trafficking of the Menkes protein are the subjects of this review. Menkes disease, a systemic copper deficiency disorder, is caused by mutations in the gene encoding the Menkes protein. The effect of these mutations on the catalytic cycle and the cell biology of the Menkes protein, as well as predictions of the effect of particular mutant MNKs on observed Menkes disease symptoms will also be discussed.     

Interactive effects of dietary silicon, copper, and zinc in the rat

A factorial rat experiment using two dietary concentrations each of copper, zinc, and silicon was conducted to identify areas in which interrelationships involving silicon may exist. The concentrations used were (mg/kg of diet): copper, 1 and 5; zinc, 2 and 12; and silicon, 5 and 270. An antagonism between silicon and zinc, whereby increases in dietary levels of either one resulted in a reduction in blood plasma concentrations of the other, was demonstrated. The depressing effect of silicon on plasma concentrations of zinc and on alkaline phosphatase occurred only in zinc-deficient rats. However, silicon had no effect on growth. Effects on aortic composition, interpreted as beneficial, accompanied increases in the silicon content of copper-deficient diets. Silicon-dependent increases in the chloroform-methanol extractable fraction of aorta closely approximated a similar response to copper. High dietary silicon increased aortic elastin in copper-deficient rats when dietary zinc was adequate. The aortic effects of silicon, while mimicking the gross effects of copper, occurred in the absence of any silicon-related changes in blood copper concentrations. Interrelationships of silicon with other elements, particularly copper and zinc, may warrant consideration in future nutritional and metabolic studies.     

Levels of Zn and Cu in the serum of a diabetic population

The zinc and copper content in serum as well as glucose and insulin was determined in 140 diabetics and compared with the results obtained from 162 healthy patients. Their mean values are significantly different. Each group was classified accordingly to the presence or not of overweight (greater than 15% relative body weight). The correlations (Pearson. Anova) between the different parameters in each group were studied. Healthy group showed a negative correlationship between the zinc and insulin levels and between the latter and the copper level, and a positive one between copper and every other parameter. Obesity only affected the zinc levels in diabetics and increased them in a statistically significant way. In this group, the copper level showed a positive correlation with the zinc level and the latter with the glucose. Owing to these results zinc and copper levels seem to be directly correlated and in diabetic patients these levels are increased mainly in the case of obesity. In healthy patients zinc levels are correlated with serum insulin, and in the diabetic groups with serum glucose.