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PubMed2004

Copper overload affects copper and iron metabolism in Hep-G2 cells.

Divalent metal transporter #1 (DMT1) is responsible for intestinal nonheme Fe apical uptake. However, DMT1 appears to have an additional function in Cu transport in intestinal cells. Because the liver has an essential role in body Cu homeostasis, we examined the potential involvement of Cu in the regulation of DMT1 expression and activity in Hep-G2 cells. Cells exposed to 10 microM Cu exhibited a 22-fold increase in Cu content and a twofold decrease in Fe content compared with cells maintained in 0.4 microM Cu. (64)Cu uptake in Cu-deficient Hep-G2 cells showed a twofold decrease in K(m) compared with cells grown in 10 microM Cu. The decreased K(m) may represent an adaptive response to Cu deficiency. Cells treated with >50 microM Cu, showed an eightfold increase in cytosolic metallothionein. DMT1 protein decreased (35%), suggesting that intracellular Cu caused a reduction of DMT1 protein levels. Our data indicate that, as a result of Cu overload, Hep-G2 cells reduced their Fe content and their DMT1 protein levels. These findings strongly suggest a relationship between Cu and Fe homeostasis in Hep-G2 cells in which Cu accumulation downregulates DMT1 activity.

Cation Transport Proteins↗
PubMed1993

Stress induction affects copper and zinc balance in calves fed organic and inorganic copper and zinc sources.

This study determined whether Cu and Zn balance was affected by feeding either Zn methionine (ZnMet) + Cu lysine (CuLys) or Zn sulfate (ZnSO4) + Cu sulfate (CuSO4) before and after stressing calves. Eight Charolais crossbred steer calves weighting 167 +/- 5 kg were randomly assigned to two treatments in a crossover experimental design. The millet hay and soybean meal diet when supplemented with the inorganic salts provided 9.2 ppm of Cu and 36.6 ppm of Zn or when fortified with the metal complexes contained 10.5 ppm of Cu and 36.6 ppm of Zn. Gentled calves were fed their respective diets for 28 d before an 18-d mineral balance trial was conducted. Collection consisted of five periods: 1) a 5-d baseline period, 2) 3 d of no Cu and Zn supplement, 3) 3 d of stress consisting of feed and water restriction and ACTH (80 IU) injections i.m. every 8 h, 4) 3 d of refeeding with no Cu and Zn supplement, and 5) 4 d of Cu and Zn repletion. Calves fed CuLys had 53% greater apparent Cu absorption and increased Cu retention (P < .05) during repletion compared with calves fed CuSO4. The 18-d mean retention of Cu from CuLys was greater (P < .05) than that from CuSO4. No differences (P > .05) in apparent absorption or retention of Zn were found between Zn sources, although during the 18-d trial mean retention was 58% higher when ZnMet was fed. Urinary Cu and Zn excretion decreased (P < .01) during stress.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2001

Zinc and copper status in ewes supplemented with sulfate- and amino acid-complexed forms of zinc and copper.

Thirty 6-yr-old Targhee ewes were randomly allotted to one of five supplemental treatments to evaluate supplementation effects on liver and fecal Zn and Cu concentrations and serum alkaline phosphatase activity: 1) control, 2) Zn complex, 3) Zn and Cu (ZnCu) complex, 4) Zn sulfate, and 5) ZnCu sulfates. Supplements were administered daily in gelatin capsules for 56 d. Liver biopsies and serum samples were collected every 14 d starting on d 0. Supplemental Zn and Cu levels were formulated to provide 90 mg/kg Zn and 10 mg/kg Cu, respectively, on a daily dry matter intake basis. Form (complex vs sulfate) x type (Zn vs ZnCu) interactions were not detected (P > 0.35). Therefore, contrast statements were used to make the following treatment comparisons: 1) control vs supplement, 2) Zn vs ZnCu, and 3) complex vs sulfate. Ewe BW at the end of the study (P = 0.09) and ewe BW change from beginning to end of the study (P = 0.07) were greater for supplemented than control ewes. Body weight and BW change did not differ between sulfate and complex (P > 0.39) or Zn- and ZnCu- (P > 0.40) supplemented ewes. Liver Cu concentrations did not differ (P = 0.41) between control and supplemented ewes. Liver Cu concentrations were higher (P < 0.10) for ewes supplemented with ZnCu than Zn and complex than sulfate forms of supplement. Liver Zn concentration tended (P = 0.13) to be higher in ZnCu than Zn-supplemented ewes. Liver and fecal Zn concentration were higher (P < 0.06) in ewes fed complex than sulfate supplements. However, serum alkaline phosphatase activity tended (P = 0.12) to be greater in ewes fed sulfate than complex supplements. Supplementing mature ewes with complexed minerals resulted in higher concentrations of Zn and Cu in the liver. In addition, supplemental Cu tended to increase concentrations of Zn in the livers of ewes; however, high levels of supplemental Zn did not negatively impact liver Cu concentrations.

Alkaline Phosphatase↗
PubMed1982

Effects of supplemental dietary copper on growth, reproductive performance and kit survival of standard dark mink and the acute toxicity of copper to mink.

Natural dark mink kit were fed a diet supplemented with 0, 25, 50, 100 or 200 ppm Cu from CuSO4 . 5H2O for 153 or 357 d. The shorter term Cu supplementation had no significant beneficial or adverse effects on mink body weight gains or hemoglobin or hematocrit concentrations, although plasma Cu concentrations were slightly elevated in the mink fed added Cu. Liver Cu concentrations were significantly increased only in the mink fed 200 ppm Cu. Liver Zn and Fe concentrations were not affected by the added Cu. Darker fur was observed in pelted males fed the higher levels of Cu. The reproductive performance of mink on the longer term Cu supplementation was not adversely affected, although greater kit mortality and reduced "litter mass" were a result of the higher Cu concentrations. The acute (21-d) ip LD50 concentrations of Cu sulfate and Cu acetate in adult mink were 7.5 and 5.0 mg/kg, respectively.

Animal Nutritional Physiological Phenomena↗
PubMed1989

Effects of source and level of copper on performance and liver copper stores in weanling pigs.

Five 28- to 33-d experiments involving 460 crossbred pigs weaned at 28 +/- 2 d of age (initial weight, 6.7 to 8.1 kg) were conducted to determine the effects of feeding high dietary levels of Cu sulfate (CuSO4) or Cu oxide (CuO) on rate and efficiency of gain and liver Cu stores of weanling pigs. The pigs were housed in groups of five to six/pen and fed a fortified, unmedicated, corn-soybean meal-dried whey basal diet (1.1% lysine, 30 ppm Cu). In Exp. 1 and 2, pigs (eight replicates) were fed the basal or the basal plus 125 or 250 ppm Cu from CuSO4 or CuO for 28 d. In Exp. 3 and 4, four replications were fed the same diets as in Exp. 1 and 2 plus two additional diets (500 ppm Cu from CuSO4 or CuO). In Exp. 5, dietary levels of 0, 125, 250, 375 or 500 ppm Cu from CuSO4 were evaluated using four replications. At the end of each experiment, the liver from one pig in each pen was collected for Cu analysis. Overall, rate and efficiency of gain were improved (P less than .01) by feeding 125 or 250 ppm Cu as CuSO4, with the 125 ppm dietary level being about 75% as effective in stimulating growth as 250 ppm. Performance of pigs was not different from controls when the highest (500 ppm) level of Cu (from CuSO4) was fed. Liver Cu increased 10- to 70-fold when 250 to 550 ppm Cu from CuSO4 was included in the feed.(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Feed↗
PubMed1987

The effect of a combination of copper and hexetidine on plaque formation and the amount of copper retained by dental plaque bacteria.

Zn++ in combination with hexetidine exerts a synergistic plaque inhibition. Studies in our laboratory on the mechanism of this effect suggested that Cu++ and hexetidine may have a similar combination effect. This hypothesis was tested in vivo on a human test panel in a double-blind crossover study. The amount of Cu++ retained by plaque bacteria in vitro was also evaluated. Seven volunteers rinsed with the solutions for 1 min twice daily for 5 days. The test solutions were H2O, 1.0 mM CuSO4, 2.0 mM hexetidine, and the last two in combination. During the test period no oral hygiene was allowed, and sucrose-containing chewing gum was used to enhance plaque formation. The plaque index scores after rinsing with the combination were significantly (p less than 0.05) lower than those of the other solutions. The effect of hexetidine on Cu++ retention in plaque bacteria was evaluated in plaque samples (n = 3) grown anaerobically overnight in PPLO medium. The bacteria were washed five times, digested in concentrated HNO3, and Cu++ determined by atomic absorption. The presence of hexetidine resulted in a significantly greater amount of Cu++ retained by bacteria at all CuSO4 concentrations. It is suggested that the nonpolar nature of the hexetidine molecule enables Cu++ bound to hexetidine to pass into the bacterial cell. Within the cell, Cu++ can interfere with bacterial metabolism, giving a reduction in plaque growth.

Adult↗
PubMed1996

Copper metabolism of holstein and jersey cows and heifers fed diets high in cupric sulfate or copper proteinate.

Eight Holstein and 8 Jersey primiparous cows (3 d postcalving) and 8 Holstein and 8 Jersey growing heifers were randomly assigned to 1 of 8 treatments in a 2 x 2 x 2 factorial arrangement to compare Cu metabolism between Holsteins and Jerseys and the bioavailabilities of Cu in Cu proteinate and CuSO4. The variables were Holstein or Jersey, Cu supplementation at 5 or 80 mg/kg of DM, and supplements of CuSO4 or Cu proteinate. Jerseys had higher hepatic Cu concentrations than did Holsteins on d 60 (346 vs. 303 micrograms/g of DM). At the high Cu supplementation, hepatic Cu increased more rapidly, and content was higher in Jerseys than in Holsteins by d 60 (520 vs. 439 micrograms/g of DM). On d 0, plasma Cu concentrations were 0.99 and 0.80 microgram/ml, and, on d 60, concentrations were 0.96 and 0.88 microgram/ml for Jerseys and Holsteins, respectively. Overall, serum ceruloplasmin oxidase activity was greater for Jerseys than for Holsteins. Jersey cows and heifers also had greater hepatic Fe (208 vs. 173 micrograms/g of DM) and lower hepatic Zn (82 vs. 91 micrograms/g of DM) than did Holstein cows and heifers at d 60. The bioavailability of Cu in Cu proteinate and CuSO4 was the same. Plasma Cu concentration and ceruloplasmin have limited value as indicators of Cu status and availability to dairy cows and heifers.

Animals↗
PubMed1999

Evaluating copper lysine and copper sulfate sources for heifers.

The effects of feeding different sources and quantities of Cu to heifers were evaluated in a 211-d experiment. Forty crossbred predominantly Brahman x Hereford heifers averaging 13.5 mo of age and 301 kg were initially depleted of Cu. The depletion diet was fed for 70 d and consisted of low Cu and high antagonist minerals, Fe, S, and Mo at 1000 mg/kg, 0.5%, and 5 mg/kg (dry basis), respectively. On d 71, heifers continued to receive the antagonistic minerals and were allotted equally to five Cu treatments: 1) control, no additional Cu source; 2) 8 mg of Cu/kg from CuSO4; 3) 16 mg of Cu/kg from CuSO4; 4) 8 mg of Cu/kg from Cu lysine; and 5) 16 mg of Cu/kg from Cu lysine. When no notable change in concentration of Cu in the liver was observed, d 169, a second diet was formulated. The heifers were fed the same Cu treatments, but S and Mo were removed and Fe was lowered to 50 mg/kg. This diet was then fed for the final 42 d of the experiment. In addition to performance, concentrations of Cu, Fe, and Zn in the plasma and liver, plasma ceruloplasmin, hemoglobin, superoxide dismutase (SOD) activity of neutrophils and lymphocytes, and a cell mediated immune response (phytohemagglutinin-P, PHA) were measured. Heifers in this study had increased growth over time, but there were no treatment differences for growth and average daily gain. Liver and plasma Cu concentrations were not greatly influenced by different supplemental Cu sources. However, compared with other treatments, Cu lysine (16 mg/kg) increased liver Cu in cattle that were deficient and tended to increase plasma Cu in animals that were marginally deficient in Cu. Iron concentrations decreased over time in liver and plasma, but there was no difference in Fe and Zn concentrations in liver and plasma among treatments. Differences in ceruloplasmin and hemoglobin concentrations were significant over time but not among treatments. The SOD activity in neutrophils did not change over time, but SOD activity of lymphocytes increased over time. For the PHA immune response test, there was no effect of time or a time by treatment interaction. These data suggest that all Cu sources were available, but Cu at 16 mg/kg from Cu lysine was more beneficial than were other sources and particularly for heifers with low Cu status.

Animal Nutritional Physiological Phenomena↗
PubMed1987

Interaction of dietary nutrient concentration and supplemental copper on chick performance and tissue copper concentrations.

An experiment was conducted with 576 female Cobb feather-sexed chicks to study the influence of methionine (MET) and selected nutrient (SN) supplementation on the performance of chicks fed high Cu levels. Day-old chicks were allotted randomly to pens for the 22-day experiment. A 3 X 2 X 2 factorial arrangement of treatments was used which included Cu at 0, 400, and 800 mg/kg, MET at 0 and .4%, and SN at 0 and 20%. A significant (P less than .001) Cu X MET X SN interaction was found for gain. Supplemental MET reversed the growth depression observed in birds fed 400 but not 800 mg/kg Cu. Additions of 400 and 800 mg/kg Cu to the basal diet depressed (P less than .001) feed consumption by 8.6 and 19.4%, respectively. Hepatic Cu concentrations increased linearly (P less than .001) with increasing dietary Cu and were not influenced (P greater than .10) by supplemental MET or SN. Liver weights increased linearly (P less than .001) with increasing dietary Cu and were higher (P less than .05) for chicks on diets supplemented with SN, but lower (P less than .05) for those with diets supplemented with MET. The interaction of MET X SN was significant (P less than .001) for serum Cu; chicks supplemented with SN in conjunction with MET had the lowest (P less than .05) serum Cu concentrations. Serum glutamic-oxaloacetic transaminase (SGOT) activity was not influenced (P greater than .10) by dietary Cu, but SN supplementation resulted in a 10% elevation (P less than .001) in SGOT activity.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1996

Dietary copper level affects copper metabolism during lipopolysaccharide-induced immunological stress in chicks.

Two experiments were conducted to examine the effect of dietary Cu level on Cu metabolism during the acute phase response in broiler chicks with adequate (Experiment 1) or deficient (Experiment 2) Cu. Diets based on cornstarch and isolated soybean protein were used to formulate a basal diet, and basal diet plus either 5, 10, or 15 mg/kg additional Cu as either CuO or CuSO4. Each diet was fed to six pens of five chicks per pen (Experiment 1) or eight pens of five chicks (Experiment 2). Half of the chicks on each diet were injected with Salmonella typhymurium lipopolysaccharide (LPS) on alternate days. In Experiment 1, LPS significantly decreased daily gain, feed intake, and feed efficiency (P < 0.01) and increased the concentration of Cu in blood plasma (P < 0.01). In the uninjected birds, adding 5, 10, or 15 mg/kg Cu as CuO or 15 mg/kg Cu as CuSO4 increased the rate of gain over that of chicks fed the basal diet. In the birds challenged with LPS, 10 mg/kg Cu as CuO increased the rate of gain and efficiency compared to those of chicks fed the basal diet. Addition of CuSO4 to the diet of chicks challenged with LPS did not affect gain, intake, or feed efficiency compared to those of chicks fed the basal diet. Ceruloplasmin levels were higher in chicks challenged with LPS than in control chicks (P = 0.03), and this difference tended to be greater in chickens fed CuO than in chickens fed CuSO4 (P = 0.07). In chicks challenged with LPS, feeding CuO at all levels and feeding CuSO4 to give 10 or 15 mg/kg Cu increased ceruloplasmin levels above that of chicks fed the basal diet. Hepatic Mn superoxide dismutase (SOD) and Cu/Zn SOD were not influenced by dietary Cu level or source or LPS. Results of Experiment 2 were similar to those of Experiment 1 except that supplemental CuSO4 and CuO gave similar increases in gain and CuSO4 was more effective at increasing ceruloplasmin levels. Chicks given supplemental Cu had higher ceruloplasmin levels following challenge with LPS than Cu-deficient chicks fed the basal diet. Apparently, Cu requirements are higher for chicks experiencing an acute phase response than for healthy chicks.

Acute-Phase Reaction↗
PubMed2000

Response of putative indices of copper status to copper supplementation in human subjects.

No sensitive functional index is currently available to assess Cu status in healthy human populations. This study evaluated the effect of Cu supplementation on putative indices of Cu status in twelve women and twelve men, aged between 22 and 45 years, who participated in a double-blind placebo controlled crossover study. The study consisted of three 6-week supplementation regimens of 3 mg CuSO4, 3 mg Cu-glycine chelate and 6 mg Cu-glycine chelate, each separated by placebo periods of equal length. Women had significantly higher caeruloplasmin oxidase activity (P < 0.001), caeruloplasmin protein concentration (P < 0.05), and serum diamine oxidase activity (P < 0.01) at baseline than men. Erythrocyte and leucocyte superoxide dismutase activity, leucocyte cytochrome c oxidase activity, and erythrocyte glutathione peroxidase activity did not respond to Cu supplementation. Platelet cytochrome c oxidase activity was significantly higher (P < 0.01), after supplementation with 6 mg Cu-glycine chelate in the total group and in women but did not change in men. Caeruloplasmin oxidase activity was significantly higher (P < 0.05), in men after supplementation with 3 mg Cu-glycine chelate, while caeruloplasmin protein concentration was significantly lower in men after supplementation with 6 mg Cu-glycine chelate (P < 0.05). Serum diamine oxidase activity was significantly higher after all supplementation regimens in the total group and in both men and women (P < 0.01). These results indicate that serum diamine oxidase activity is sensitive to changes in dietary Cu intakes and may also have the potential to evaluate changes in Cu status in healthy adult human subjects.

Adult↗
PubMed1988

A clinical study comparing the performance of three IUCDs (Multiload Copper 250, Copper T 200, and Lippes Loop C).

300 Women at Cairo's Al-Zahraa Hospital, using 1 of 3 different IUds (CuT 200, multiload Cu250, and Lippes loop size C) to compare the rate of complications encountered among the 3 groups. Failed insertion was highest among the Cu T200 group with a rate of 3.24/HWY, hundred woman years. Accidental pregnancy was highest among that group as well. With a rate 2.16/HWY compared with 1.10/HWY among the ML Cu250 group and none among the Lippes Loop group. The spontaneous expulsion rate was the same for the Cu T200 and Lippes Loop, i.e., the rate was 4.32/HWY and 4.44/HWY for the 2 groups, respectively, compared with 1.10.HWY for the ML Cu250. The highest removal rate because of pain or bleeding was encountered among the ML Cu250 users, i.e., the rate was 4.41.HWY compared with 3.33/HWY among the Lippes Loop group and 1.08/HWY for the Cu T200 group. The overall failure rate was 8.82/HWY for ML Cu250, 11.89/HWY for Cu T200, and 11.10/HWY for the Lippes Loop group.

Africa↗