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.     

Effects of copper aspirinate and aspirin on tissue copper,zinc, and iron concentrations following chronic oral treatment in the adjuvant arthritic rat

Concentrations of copper, zinc, and iron were analyzed and compared in a number of tissues of adjuvant arthritic rats following 22 d of chronic treatment (per os) with either vehicle, aspirin or copper aspirinate, at doses of 100 mg/kg, 200 mg/kg, or 400 mg/kg. Such chronic treatment resulted in a negative balance in copper, zinc, and iron in many tissues. Among the tissues examined, liver and kidney exhibited the greatest changes in metal concentrations; brain and skeletal muscle exhibited the least. Arthritis-induced changes in the concentrations of all three metals in the liver were reversed upon treatment with aspirin. Treatment with copper aspirinate, on the other hand, resulted in an extremely high accumulation of copper in the liver. Arthritis-induced changes in copper, zinc, and iron concentrations in the pancreas and copper concentration in the plasma were generally not reversed upon treatment with either aspirin or copper aspirinate. Among the three metals examined, the degree of change observed as a result of drug treatments was greatest for iron and least for zinc. Finally, it appeared that the effects of aspirin and copper aspirinate on tissue metal concentrations were independent of the antiarthritic effects of these compounds.     

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.     

Inhibition of mugineic acid-ferric complex uptake in barley by copper, zinc and cobalt

The interfering effects of copper, zinc, and cobalt on the uptake of mugineic acid-ferric complex were studied in barley ( Hordeum vulgare , cv. Minorimugi) grown in nutrient solution. Short-term uptake experiments of 3 h were performed utilizing both ionic and mugineic acid-complex forms of each metal at two different concentrations. Copper was most effective in decreasing iron uptake when added in an ionic form at either concentration. The inhibition order at higher concentrations followed Cu(II) > Zn(II) ≥ Co(II), Co(III), which is consistent with the stability constants of these metal complexes with mugineic acid. The displacement of iron from its mugineic acid complex by these metals is suggested as a probable explanation for the decreased iron uptake. The inhibitory effect of metal complexes with mugineic acid on iron uptake was only found in cases with higher concentrations of Cu(II) and Zn(II) complexes. Deformation of the specific iron transport system in the plasma membrane due to their adsorption may be responsible for this effect.     

The tissue disposition and urinary excretion of cadmium, zinc, copper and iron, following repeated parenteral administration of cadmium to rats.

The effect of repeated parenteral administration of cadmium (0.75, 1.5 and 3.0 mg/kg) on tissue disposition and urinary excretion of cadmium, zinc, copper and iron has been studied in the male rat. Cadmium, zinc and copper accumulated in liver and kidney, but the concentration of iron did not alter significantly. The kidney weight relative to body weight showed a dose-related increase in weight of 25--65%. Excretion of cadmium in the urine increased directly with dosage and the increase was most significant when kidney damage had probably occurred. Administration of cadmium also resulted in dose-related increases in the urinary excretion of zinc, copper and iron. The cadmium concentration of blood increased with dosage of cadmium, and the plasma concentrations of zinc and copper were also raised but plasma iron concentration was diminished.     

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.     

Kinetics and mechanism of tolerance induction on acclimation ofVillorita cyprinoides (Hanley) to copper and zinc

The typical euryhaline clamVillorita cyprinoides (Hanley) was acclimated to copper and zinc at salinity 13 × 10⁻³ and < 1 × 10⁻³ (fresh water). Acclimation enhanced the lethal tolerance, as denoted by dose-survival curves, which was more pronounced after zinc acclimation. In fresh water copper acclimation sensitized the organisms. The copper accumulation trend was significantly changed consequent to metal acclimation, especially after zinc acclimation, indicating some tissue metal regulatory effect. Acclimation to copper equiped the organism to survive for longer periods with increased body burden of copper, while zinc acclimation supressed the uptake of the more toxic ion copper. The earlier report of increased uptake of zinc by this organism during combined exposure with copper is corelated in the present context. The role of metallothionein like protein in providing protection against metal toxicity, the environmental implication of acclimation phenomena are indicated     

Comparative study of copper and zinc metabolism in cattle and camel

At an experimental farm, five camels and five cows were fed a similar basal diet for 6 mo. They received oral trace element supplementation for 3 mo (day 22–112). This supplementation included zinc, copper, selenium, managanese, iodine, and cobalt, and corresponded to twice the requirements generally recommended for cows. Plasma copper and zinc concentrations were significantly lower in the camels (61 μg/100 mL for copper and 38 μg/100 mL for zinc) than in the cows (111 and 83, respectively). The supplementation had no effect on plasma zinc concentration in the camels in spite of the low observed values in this species. Liver copper concentration at the beginning of the trial was lower in the camels (9 ppm) than in the cows (35 ppm), and stayed at lower levels during the entire supplementation period. There was no clear difference in fecal excretion of copper and zinc between the camels and the cows. The results suggested that trace element requirements are lower in camels than in cows and that camels regulate their plasma zinc concentration at a very low level (<40 μg/100 mL).     

Zinc, copper, and iron metabolism during porcine fetal development

Zinc, copper, and iron levels in maternal and fetal pig tissues and fluids were measured starting on d 30 of gestation and continuing to term (d 114) at 10-d intervals. Fetal hematocrit increased from a low of 19% on d 30 to 32% by d 50, after which it remained above 30% to term. Amniotic fluid zinc, copper, and iron all reached maximal levels by d 60 of gestation. Maternal serum zinc levels fluctuated little during gestation, but fetal serum zinc concentration was significantly elevated above maternal levels during the second trimester. Fetal serum copper levels were significantly lower than maternal values throughout gestation and this was also the case for ceruloplasmin oxidase activity. Maternal serum iron reached its lowest level by d 80 of gestation when rate of transfer of iron to the developing fetuses was high. Fetal serum iron declined throughout gestation, reaching its lowest level on d 100. In general, fetal liver concentrations of zinc, copper, and iron were higher than the corresponding maternal values throughout gestation. Distinct increases were noted for fetal hepatic zinc and copper concentrations during the second trimester of pregnancy and these were accompanied by increases in cytosolic and metallothionein-bound zinc and copper levels. Maternal hepatic iron declined during the second trimester, reaching its lowest point on d 80, indicative of the shunting of maternal iron reserves to fetal tissues. Fetal kidney metal levels did not demonstrate any distinctive developmental patterns with respect to zinc, copper, or iron concentrations, but a general accumulation of each metal was observed as gestation progressed. The results of this study highlight some of the distinct changes occurring in the metabolism of zinc, copper, and iron in both maternal and fetal tissues and fluids during gestation in the pig. Mention of a trade name, proprietary product, or specific equipment does not constitute a guarantee or warranty by the U.S. Department of Agriculture and does not imply its approval to the exclusion of other suitable products.     

The combined effect of high iron and zinc intake on copper status in rats

The interactions between copper, zinc, and iron intake in rats were investigated with regard to copper status. Weanling male rats were fed purified diets containing two levels of each of the three elements in a 2³ factorial design. The added amounts of copper, zinc, and iron in the diets were 5, 12, and 35 mg/kg feed or were 10 times as high. After feeding on the experimental diets for 4 wk, the rats were killed and copper concentrations in plasma and organs measured. Plasma copper concentration was lowered by high zinc and iron intakes but this was seen only in the rats fed the normal-copper instead of the high-copper diets. In essence, the effects of zinc and iron were additive. Neither in rats fed the normal-copper diets nor in those fed the high-copper diets did extra iron or zinc intake alter copper concentrations in liver, spleen, kidney, and tibia.     

Ascorbic acid feeding of rats reduces copper absorption,causing impaired copper status and depressed biliary copper excretion

The feeding of diets enriched with ascorbic acid (10 g/kg) to rats has previously been shown to lower plasma and liver copper concentrations. The present studies corroborate this. We hypothesized that ascorbic acid initially reduces copper absorption, this effect being masked later by the stimulatory effect on copper absorption of the impaired copper status. We also hypothesized that the impaired copper status as induced by ascorbic acid feeding is followed by a diminished biliary excretion of copper in an attempt to preserve copper homeostasis. Our hypotheses are supported by the present studies. Ascorbic acid feeding initially reduced apparent copper absorption, and in the course of the experiment this effect tended to turn over into a stimulatory effect. Copper deficiency, as induced by feeding a diet containing 1 mg Cu/kg instead of 5 mg Cu/kg, systematically increased copper absorption. Biliary excretion of copper in rats given ascorbic acid was unaffected initialy but became depressed after prolonged ascorbic acid feeding. A similar time course was seen for fecal endogenous copper excretion that was calculated as the difference between true and apparent copper absorption. Copper deficiency systematically reduced biliary copper excretion and fecal endogenous copper loss.     

Trypanosoma evansi: Levels of copper, iron and zinc in the bloodstream of infected cats

The aim of this study was to evaluate the concentrations of copper, iron and zinc in blood serum of cats experimentally infected with Trypanosoma evansi. Animals were divided into two groups: control and infected with T. evansi. The animals were infected with 10⁸ trypomastigotes each and parasitemia was estimated daily for 56 days by microscopic examination of smears. Hematological and biochemical parameters were evaluated for monitoring of the disease. Serum metal levels were determined in blood samples collected at days 7, 28 and 56 of the experiment. Inductively coupled plasma optical emission spectrometry was used to measure the levels of copper, iron and zinc. Significant differences were observed among groups (P < 0.05). Increased levels of copper and decreased iron and zinc levels were observed. A decrease in the number of red blood cells was also observed 7 days after inoculation. Biochemical parameters were not altered. Therefore, the infection by T. evansi might alter the serum metal levels, causing metabolic disturbances in cats.     

Influence of food ration, copper exposure and exercise on the energy metabolism of common carp (Cyprinus carpio)

The present study was conducted to extend the understanding of the combined physiological effects of different food rations in combination with sublethal levels of copper in common carp (Cyprinus carpio). Fish acclimated to low (0.5% body weight) and high (5% body weight) food rations were exposed to 1 microM copper for a period of 28 days and kept for a further 14 days in copper free water to examine their recovery. Measurements of oxygen consumption, ammonia excretion and ammonia accumulation in plasma and muscle were done at various time intervals during the experimental period. Overall, oxygen consumption and ammonia excretion rates were significantly affected by food ration in both copper free and copper exposed fish. Additional challenges, such as copper exposure and/or exercise, significantly increased plasma and muscle ammonia in the fish fed a high food ration. Muscle ammonia levels in general responded slower (first increase after 72 h) and recovered within 2 weeks of exposure. There was a significant correlation between plasma ammonia levels, muscle ammonia levels and ammonia excretion rates. Influence of copper in terms of ammonia excretion and plasma ammonia accumulation was observed in high ration fish but low ration fish remained unaffected. This clearly indicates that ammonia metabolism was significantly influenced by copper in this group of fish showing that during unfavourable environmental conditions a high amount of food supply may turn deleterious to fish.     

Characterization and crystal structure of zinc azurin, a by-product of heterologous expression in Escherichia coli of Pseudomonas aeruginosa copper azurin.

Azurin*, a by-product of heterologous expression of the gene encoding the blue copper protein azurin from Pseudomonas aeruginosa in Escherichia coli, was characterized by chemical analysis and electrospray ionization mass spectrometry, and its structure determined by X-ray crystallography. It was shown that azurin* is native azurin with its copper atom replaced by zinc in the metal binding site. Zinc is probably incorporated in the apo-protein after its expression and transport into the periplasm. Holo-azurin can be reconstituted from azurin* by prolonged exposure of the protein to high copper ion concentrations or unfolding of the protein and refolding in the presence of copper ions. An X-ray crystallographic analysis of azurin* at 0.21-nm resolution revealed that the overall structure of azurin is not perturbed by the metal exchange. However, the geometry of the co-ordination sphere changes from trigonal bipyramidal in the case of copper azurin to distorted tetrahedral for the zinc protein. The copper ligand Met121 is no longer co-ordinated to zinc which adopts a position close to the carbonyl oxygen atom from residue Gly45. The polypeptide structure surrounding the metal site undergoes moderate reorganization upon zinc binding. The largest displacement observed is for the carbonyl oxygen from residue Gly45, which is involved in copper and zinc binding. It moves by 0.03 nm towards the zinc, thereby reducing its distance to the metal from 0.29 nm in the copper protein to 0.23 nm in the derivative.     

Magnesium,calcium, copper,and zinc in meals

Meals of 12 diets were prepared from conventional foods with precautions against contamination by metallic elements because of epidemiologic associations between ischemic heart disease and the metabolism of magnesium, calcium, copper, and zinc. Magnesium, calcium, copper, and zinc were measured by atomic absorption spectrometry with satisfactory accuracy and precision. The mean daily amount of copper in the diets was less than the apparent adult requirement. Mean amounts of magnesium and zinc were close to apparent requirements; however, adults consuming amounts less than one standard deviation below these means may be depleting body stores and be at risk of pathology. Magnesium and copper were highly correlated (r=0.849,P=0.0001) in meals. This and other significant correlations probably will prevent the relationships of these elements to ischemic heart disease from being elucidated by epidemiology. Metabolic experiments will be necessary to differentiate among several hypotheses.     

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.     

Comparative strategies of heavy metal accumulation by crustaceans: zinc,copper and cadmium in a decapod,an amphipod and a barnacle

This study investigates the comparative strategies of accumulation under standardised laboratory conditions of the essential metals zinc and copper, and the non-essential metal cadmium by three crustaceans of different taxa; vizPalaemon elegans Rathke (Malacostraca: Eucarida: Decapoda),Echinogammarus pirloti (Sexton & Spooner) (Malacostraca: Peracarida: Amphipoda) and the barnacleElminius modestus Darwin (Cirripedia: Thoracica).The decapodP. elegans regulates body zinc concentrations to a constant level (ca. 79 µg Zn g^–1) over a wide range of dissolved metal availabilities until regulation breaks down at high Zn availabilities and net accumulation begins. The amphipodE. pirloti accumulates zinc at all dissolved zinc concentrations but at a low net rate such that the accumulation strategy approaches that of regulation. The barnacleE. modestus accumulates zinc to high body concentrations with no significant excretion of accumulated zinc. In the case of copper,P. elegans similarly regulates body copper concentrations to a constant level (ca. 129 µg Cu g^–1) over a range of dissolved copper availabilities until regulation breaks down at high copper concentrations. Both the amphipodE. pirloti and the barnacleE. modestus on the other hand accumulate copper at all dissolved copper exposures with no evidence of regulation. All three crustaceans accumulate the non-essential metal cadmium at all dissolved cadmium concentrations without regulation.Heavy metal accumulation strategies therefore vary between crustacean taxa and between metals. Uptake rates for zinc and cadmium have been estimated for the three crustaceans and can be interpreted in terms of cuticle permeability and way of life of each crustacean. Examination of these uptake rates provides an insight into possible reasons behind the adoption of particular metal accumulation strategies.     

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.     

Biological significance of cimetidine sulfoxide complexes with copper(II) and zinc(II) ions during cimetidine treatment

Following a recent investigation into cimetidine interactions with copper(II) and zinc(II), the present work deals with the study of coordination equilibria relative to the main metabolite of this drug, i.e. cimetidine sulfoxide, with the same metal ions under physiological conditions.Computer simulations were run on the basis of the corresponding complex stability constants, in order to assess the extent to which cimetidine sulfoxide may affect copper(II) and zinc(II) plasma distributions during long term cimetidine therapy.No significant effect of this kind can be expected from cimetidine sulfoxide for plasma concentrations corresponding to usual therapeutic levels of the patent molecule, even for patients presenting with impaired renal function.     

Urinary excretion of mercury, copper and zinc in subjects exposed to mercury vapour

The excretion of mercury, copper and zinc in urine, and mercury in whole blood andplasma, was determined in 40 chloralkali workers exposed to mercury vapour and 40age-matched referents. The Hg concentrations in whole blood, plasma and urine werehigher in the exposed group (35 nmol l, 30 nmol l,and 11.5 nmol mmol creatinine, respectively) in comparison with thereference group (15 nmol l, 6.3 nmol l, and 1.8nmol mmol creatinine, respectively). The urinary copper excretionwas similar in the two groups, while U-Zn excretion was significantly higher (P = 0.04)in the exposed group, median 0.83 mmol mmol creatinine versus 0.76mnmol mmol creatinine in the reference group. In a subgroup of exposedworkers with current U-Hg above 11.5 nmol lmmolcreatinine (20 mg g creatinine) the medianU-Zn was 1.1 mmol mmol creatinine. In both groups smokers had highU-Zn levels than non smokers. When both U-Hg and smoking were taken into account in alinear regression model, there was a significant association between U-Hg and U-Zn inthe combined group of exposed and referents (P = 0.002). This study indicates thatmercury exposure in humans, as in animals, causes increased urinary excretion of zinc.The mechanisms may be induced synthesis of metallothionein in the kidneys, displacementof Zn from preexisting metallothionein by Hg, or a decreased reabsorption of zinc in thekidneys owing to a slight tubular dysfunction.