A moderately high intake compared to a low intake of zinc depresses magnesium balance and alters indices of bone turnover in postmenopausal women.

OBJECTIVES: To determine whether moderately high or low intakes of zinc adversely affect the copper status of postmenopausal women to result in unfavorable changes in calcium and magnesium metabolism and other indicators of bone turnover. DESIGN: After a 10-day equilibration period in which the diet provided 31.5 micromol (2 mg) Cu and 137.7 micromol (9 mg) Zn/8.4 MJ (2000 kcal), the subjects were randomly divided into two groups, with one group fed the basal diet supplemented to provide 15.7 micromol (1 mg) Cu/8.4 MJ, and the other group fed the same diet supplemented to provide 47.2 micromol (3 mg) Cu/8.4 MJ. After equilibration, both groups were fed the basal diet with no zinc supplemented (provided 45.9 micromol [3 mg] Zn/8.4 MJ) for 90 days; this was followed by another 10-day equilibration period before the basal diet was supplemented with zinc to provide 811 micromol (53 mg)/8.4 MJ for 90 days. SETTING: The metabolic unit of the Grand Forks Human Nutrition Research Center, Grand Forks, ND, USA. SUBJECTS: A total of 28 postmenopausal women recruited by advertisement throughout the United States of America. Among them, 25 women (64.9+6.7 y) completed the study; 21 as designed. RESULTS: The moderately high intake compared to the low intake of zinc increased the excretion of magnesium in the feces and urine, which resulted in a decreased magnesium balance. In the women fed low dietary copper, plasma osteocalcin was higher during the low-zinc than high-zinc dietary period. The urinary excretion of N-telopeptides was increased and the serum calcitonin concentration was decreased by high dietary zinc regardless of dietary copper. CONCLUSIONS: A moderately high intake of zinc (811 micromol/day; 53 mg/day) did not induce changes in copper metabolism that resulted in unfavorable changes in bone or mineral metabolism. However, low dietary zinc (45.9 micromol/day; 3 mg/day) apparently resulted in undesirable changes in circulating calcitonin and osteocalcin. As a moderately high intake of zinc decreased magnesium balance, further study of the possibility that a high intake of zinc is a health concern for individuals consuming less than the recommended amounts of magnesium is warranted.

Low dietary zinc alters indices of copper function and status in postmenopausal women.

OBJECTIVES: To better define the relationship between dietary zinc and copper for humans so that sound recommendations for intakes of these elements can be made. METHODS: A study was conducted to ascertain the effect of moderately excessive and deficient intakes of zinc on copper metabolism and use in humans fed low and luxuriant amounts of copper. Twenty-one postmenopausal women housed in a metabolic unit completed the study as designed. After a 10-d equilibration period in which they were fed a diet providing 31.5 micromol (2 mg) Cu and 91.8 micromol (9 mg) Zn/8.4 MJ (2000 kcal), the women were divided into two groups. One group was fed a diet containing 15.7 micromol (1 mg) Cu/8.4 MJ (2000 kcal), and the other group was fed a diet containing 47.2 micromol (3 mg) Cu/8.4 MJ (2000 kcal). After equilibration, both groups were fed the basal diet providing 45.9 micromol (3 mg) Zn/8.4 MJ (2000 kcal) for 90 d; this was followed by another 10-d equilibration period before dietary zinc was increased to 811 micromol (53 mg)/8.4 MJ (2000 kcal) for 90 d. RESULTS: The women were in positive copper balance only when the diet provided 47.2 micromol (3 mg) Cu and 811 micromol (53 mg) Zn/d. Immunoreactive ceruloplasmin concentrations and platelet cytochrome-c oxidase activity on a platelet number basis were significantly lower and the ratio between enzymatic and immunoreactive ceruloplasmin was significantly higher during low dietary than during high dietary zinc intake. Serum cholesterol was higher in subjects fed 15.7 micromol (1 mg) Cu/d than in those fed 47.2 micromol (3 mg) Cu/d. Total and low-density lipoprotein cholesterol concentrations decreased with zinc supplementation. Whole-blood glutathione concentration and erythrocyte glutathione peroxidase activity were lower during high than during low dietary zinc intake. CONCLUSIONS: The findings indicate that an inadequate intake of zinc (45.9 micromol/d; 3 mg/d) was more effective than a moderately high intake of zinc (811 micromol/d; 53 mg/d) in inducing changes associated with a decreased copper status in postmenopausal women. Furthermore, the findings indicate that copper status indicators might be useful in evaluating changes in zinc status in humans, and an intake of 15.7 micromol (1 mg)/d of copper may be inadequate for postmenopausal women.

Interactions among dietary fat, mineral status, and performance of endurance athletes: a case study.

In a pilot study, performance measures and mineral metabolism were assessed in 3 male endurance cyclists who consumed isoenergetic, isonitrogenous diets for 28-day periods in a randomized, crossover design in which dietary carbohydrate, polyunsaturated, or saturated fat contributed about 50% of daily energy intake. Peak aerobic capacity [62 ml/(kg á min)] was unaffected by diet. Endurance capacity at 70-75% peak aerobic capacity decreased with the polyunsaturated fat diet. Copper retention tended to be positive only with saturated fat. Less iron and zinc were retained (intake - losses), and fecal losses of these minerals increased with the polyunsaturated fat. Blood biochemical measures of trace element nutritional status were unaffected by diet, except serum ferritin, which tended to decrease during consumption of the polyunsaturated fat diet. These preliminary results suggest that diets high in polyunsaturated fat, particularly linoleic acid, impair absorption and utilization of iron and zinc, and possibly magnesium, and may reduce endurance performance.

Changes in dietary zinc and copper affect zinc-status indicators of postmenopausal women, notably, extracellular superoxide dismutase and amyloid precursor proteins.

BACKGROUND: Zinc is an essential trace element for human health and well-being; however, methods currently available for the assessment of zinc status in humans are unsatisfactory. OBJECTIVE: The objective was to critically evaluate the use of various indicators of zinc status in humans in a controlled metabolic ward study. DESIGN: Indicators of zinc status were measured in 25 healthy postmenopausal women aged 64.9 +/- 6.7 y. After a 10-d equilibration period, volunteers consumed a diet with either a low (1 mg/d; n = 12) or a high (3 mg/d; n = 13) copper content based on a total energy content of 8.4 MJ. They received the same amount of copper throughout the study. Both groups were fed the basal diet (3 mg Zn/d) with no zinc supplement for one 90-d period, and the diet supplemented with 50 mg Zn/d for another 90-d period. RESULTS: Zinc supplementation significantly increased (P < 0.0001) extracellular but not erythrocyte superoxide dismutase activity. This increase was more apparent when subjects were fed the low-copper diet. Zinc supplementation in combination with the low-copper diet significantly decreased (P < 0.01) amyloid precursor protein expression in platelets. Other indicators of zinc status that were significantly elevated after zinc supplementation were as follows: plasma zinc and free thyroxine concentrations and mononuclear 5'-nucleotidase activity. CONCLUSION: The measurement of serum extracellular superoxide dismutase activity may be useful as a marker for the functional assessment of zinc status in humans.

The interaction between dietary fructose and magnesium adversely affects macromineral homeostasis in men.

OBJECTIVE: Studies with rats have found that an interaction between fructose and magnesium affects macromineral metabolism; high dietary fructose significantly increased kidney calcification in both male and female rats, particularly when dietary magnesium was low. This study tests the hypothesis that an interaction between dietary fructose and magnesium adversely affects macromineral homeostasis in men. METHODS: Eleven men aged 22 to 40 years were fed a mixed, Western diet for four 42-day dietary periods in which dietary magnesium was either approximately 170 or 370 mg/day and dietary fructose was either 4% or 20% of energy. A decaffeinated beverage containing high fructose corn syrup replaced cornstarch, bread and rice in the low fructose diet to give the high fructose diet. RESULTS: High dietary fructose significantly (p<0.01) increased magnesium balance during both low and high dietary magnesium intakes. Ultrafilterable and ionized serum magnesium also apparently were related to magnesium and fructose intakes; they were higher when fructose was fed and when Mg intakes were high. High fructose depressed calcium balance: the effect tended to be more marked when dietary Mg was low. High dietary fructose also significantly (p<0.005) decreased phosphorous balance. Urinary phosphorous losses were significantly (p<0.001) higher when high dietary fructose was fed. High dietary fructose also increased the concentration of serum alkaline phosphatase (p<0.005). CONCLUSION: These findings indicate that dietary fructose adversely affects macromineral homeostasis in humans and suggest further studies to see if a high fructose diet coupled with low dietary magnesium and marginal calcium leads to bone loss.

Copper intake and assessment of copper status.

The diagnosis of marginal copper deficiency has not been perfected despite an increased understanding of the physiologic roles of copper. The use of nonstandardized procedures and the effects of factors other than copper nutriture have impeded identification of an ideal indicator of copper nutritional status in humans. A review of studies of experimental copper deprivation conducted in adult humans over the past 12 y indicated that between 1.0 and 1.25 mg Cu/d is needed by adults for copper maintenance for periods of up to 6 mo and that < or = 2.6 mg Cu/d for periods of up to 42 d is not sufficient for recovery from copper deprivation. Copper-containing enzymes in blood cells, such as erythrocyte superoxide dismutase and platelet cytochrome-c oxidase, may be better indicators of metabolically active copper and copper stores than plasma concentrations of copper or ceruloplasmin because the enzyme activities are sensitive to changes in copper stores and are not as sensitive to factors not related to copper nutriture.

Chromium supplementation and resistance training: effects on body composition, strength, and trace element status of men.

The effects of 8 wk of daily chromium supplementation (3.3-3.5 mumol as chromium chloride or chromium picolinate) or placebo (0.1 mumol Cr) and weight training were examined in 36 men in a double-blind design. Strength, mesomorphy, fat-free mass, and muscle mass increased with resistance training independently of chromium supplementation (P < 0.0001). Protein, magnesium, zinc, copper, and iron intakes equalled or exceeded the recommended dietary allowance (RDA) or estimated safe and adequate daily dietary intake (ESADDI) during training and did not change significantly from pretraining intakes (P > 0.05). Chromium supplementation increased the serum chromium concentration and urinary chromium excretion without a difference as a result of the chemical form of chromium (P < 0.05). Resistance training was associated with a significant decrease (P < 0.05) in serum ferritin, total-iron-binding capacity, transferrin saturation, the ratio of enzymatic to immunoreactive ceruloplasmin, and plasma copper, independently of chromium supplementation. However, transferrin saturation was decreased more with chromium picolinate supplementation (24%) than with chromium chloride or placebo (10-13%). Compared with pretraining values, urinary magnesium excretion increased (P < 0.05) and urinary zinc output tended to decrease during the first 4 wk of resistance training and then returned to baseline values for the final 4 wk, which suggests an adaptation in mineral excretion in response to weight training. These findings suggest that routine chromium supplementation has no beneficial effects on body- composition change or strength gain in men. Whether chromium supplementation of individuals with diminished chromium nutriture facilitates propitious changes in body structure and function remains to be determined.

Effects of a diet low in copper on copper-status indicators in postmenopausal women.

To study the effects of low copper intake in older individuals, 12 postmenopausal women, aged 63.1 +/- 8.8 y, were fed a diet containing 9 micromol (0.57 mg) Cu/d for 105 d, followed by a copper-repletion period of 35 d during which the diet was supplemented with 31.5 micromol (2.0 mg) Cu/d. Plasma copper and ceruloplasmin did not change significantly during copper depletion but ceruloplasmin decreased during copper repletion. Erythrocyte superoxide dismutase activity dropped significantly during low copper intake from 3450 to 2600 U/g hemoglobin, but did not increase during copper repletion. Platelet cytochrome c oxidase activity changed significantly (P<0.0001) from 1740 to 810 U/g protein during copper depletion, then increased to 1000 U/g protein during copper repletion. Erythrocyte glutathione peroxidase activity responded similarly. Clotting factor VIII activity increased significantly during copper depletion, then dropped during copper repletion. Low copper intakes did not induce the changes in serum cholesterol and hematology generally found in copper-deficient animal models. These results indicate that a paradigm shift may be needed in evaluating copper status in adult humans. Sensitive indicators of copper include functional activities of platelet cytochrome c oxidase, platelet copper, glutathione peroxidase, and clotting factor VIII. Plasma copper, ceruloplasmin, and cholesterol are relatively insensitive indicators. Also, the recovery from mild copper depletion may require more aggressive intervention tha 2 mg Cu/d for 35 d.

Assessment of copper nutritional status.

Despite increased interest in the role of copper deficiency in clinical problems and an increased understanding of the physiological roles of copper, the diagnosis of a marginal deficiency has not been perfected. The use of non-standardized procedures and the effects of factors other than copper nutriture have impeded identification of the "ideal" indicator of copper nutritional status in adult humans. The specific activity of copper enzymes, or of copper-containing enzymes in blood cells, such as erythrocyte superoxide dismutase and platelet or leukocyte cytochrome c oxidase, may be a better indicator of metabolically active copper stores than the serum concentration of copper or ceruloplasmin, because the enzyme activities are sensitive to changes in copper stores and are not as sensitive to factors not related to copper nutriture. A single index, such as serum copper concentration, is inadequate for assessing the total body copper nutriture of an individual and must be supported by corroborating evidence.

Biochemical assessment of vitamins and trace metals.

The choice of nutritional assessment tests must be made with care, as the available tests may reflect recent dietary intakes or body tissue stores to different degrees. The interpretation of nutritional assessment tests is complicated by the dynamic nature and broad range of nutrient intake and metabolism and by a relative lack of age- and sex-specific reference ranges. The latter problem can be minimized by the use of function-based methods, such as the measurement of vitamin-dependent red cell enzyme activities to assess B vitamin status; however, few suitable function-based methods are now available, and more research in this area is needed. Because calibration standards and quality control materials may not be readily available, laboratory analysts planning nutritional assessment testing must be especially careful in establishing the credibility of assays and the quality control program. Recent advances in analytic methodology include the specificity offered by HPLC techniques and methodologies that provide simultaneous determination of more than one nutrient (or form of nutrient) in a single procedure. Examples of the latter include HPLC methods that provide B vitamin or fat-soluble vitamin profiles, and emission spectrometry techniques that provide trace metal profiles. Further work in providing useful nutritional assessment profiles and convenient automated methods is needed.

Assessment of copper status: effect of age and gender on reference ranges in healthy adults.

We measured major indices related to copper nutritional status in 55 men and 86 women between ages 20 and 83 years who were in apparent good health. Plasma copper concentrations and both immunoreactive and enzymatically measured ceruloplasmin were significantly higher in women than in men and were higher in women taking oral contraceptives. Plasma copper, immunoreactive ceruloplasmin, and cytochrome-c oxidase in platelets and mononucleated leukocytes tended to increase with age. The ratio of enzymatic to immunoreactive ceruloplasmin, erythrocyte superoxide dismutase, and 67Cu uptake by erythrocytes were not significantly affected by either age or gender. Thus, factors other than copper nutriture--such as age, gender, and hormone use--need to be considered when using many of these indicators to evaluate copper nutritional status.

Homeostatic control of zinc metabolism in men: zinc excretion and balance in men fed diets low in zinc.

Zinc metabolism was studied in 11 men. The study began with 28-d equilibration when dietary zinc was 159 mumol/d (X), followed by 35-d periods when the diet contained 21.9 (I), 37.5 (II), 51.6 (III), or 67.8 (IV) mumol Zn/d in random order, and ended with 35 d with X. The diet was conventional foods and egg white protein. Zinc balance, including surface and semen losses, was zero only during I. Semen zinc was unaffected by diet. Plasma zinc dropped to 0.44 and 0.49 mumol/L in two subjects during I and was significantly decreased during I compared with X (P < 0.0002). Urinary zinc declined with decreasing zinc intake. A combination of urinary and plasma zinc criteria from Baer and King (Am J Clin Nutr 1984; 39:556-70) could be used to distinguish stages of zinc deficiency. By these criteria, no subjects were deficient during IV, one was marginally deficient during III, three were marginally deficient during II, and seven were deficient during I.

Effects of age and sex on copper absorption, biological half-life, and status in humans.

Healthy, free-living men and women aged 20-83 y (n = 127) were studied to determine the effects of age and sex on copper absorption, biological half-life (BH), and status. Copper absorption was greater in women (71%) than in men (64%) aged 20-59 y (P = 0.02), but did not differ in men and women aged 60-83 y. BH of 67Cu ranged from 13 to 33 d and differed between men and women aged 20-59 y (P = 0.006), but not between men and women aged 60-83 y. Plasma copper, enzymatic ceruloplasmin (Cp), and immunoreactive (RID) Cp were significantly higher in women than in men (P < 0.005), but superoxide dismutase (SOD) and in vitro 67Cu uptake by red blood cells did not differ. Plasma copper, RID Cp, and cytochrome oxidase in platelets and mononuclear cells were significantly affected by age (P < 0.005). Oral contraceptives elevated plasma copper, enzymatic Cp, and SOD activity but not copper absorption and BH in women aged 20-39 y. Copper intake from self-selected diets was 0.9-1.2 mg/d for women and 1.2-1.3 mg/d for men, but net copper absorption (micrograms Cu.kg body wt-1.d-1) did not differ. Thus, dietary copper intake requirements may differ between men and women.

Short- and long-term variability of selected indices related to nutritional status. II. Vitamins, lipids, and protein indices.

Five free-living women (ages 23-38 years) who consumed a self-selected diet and five women (ages 23-44 years) residing in a metabolic unit who were fed constant diet were assessed for variation in vitamin and general chemistry indices. Blood was drawn from these women once a month for five months, once a week for five weeks, and once a day for five days to assess analytical and biological variability of the indices. Analytical variability was determined by concurrently analyzing control samples prepared from plasma and serum pools. All samples were analyzed in duplicate. Of the measured indices, vitamins and lipids seemed to be the most variable. Diet had a significant effect only on ascorbic acid. We were unable to show any seasonal change for these analytes. Estimations of analytical variability, along with estimates of biological variability, and knowledge of dietary practices are essential when interpreting differences in analytes.

Nutrition status and brain function in aging.

Biochemical indices of nutrition status assessed in 28 healthy persons aged greater than 60 y were related to cognitive performance and electroencephalographic (EEG) indices of neuropsychological function. Performance data were most frequently related to indices of nutrition status when tasks were demanding. Numerous correlations were also found between EEG indices and indices of thiamin, riboflavin, and iron nutriture. Certain observations, such as a decrement in alpha-wave activity in the EEG of subjects with low thiamin status, suggest that subtle neuropsychological impairment can occur in association with mild deficits in nutrition status. Other findings indicate that EEG frequency responses of older subjects with high iron status are similar to those of younger persons; however, these data are more difficult to interpret. The results suggest that further research on nutrition and neuropsychological function will lead to a better understanding of the role of nutrition in maintaining the functional integrity of the aging brain.

Manganese content of the cellular components of blood.

We measured the manganese content of whole blood, plasma, platelets, mononucleated cells, polymorphonucleated cells, and erythrocytes. Platelets and blood cells were separated from whole blood by use of discontinuous gradients of colloidal polyvinylpyrrolidone-coated silica (Percoll), and their manganese content was measured by Zeeman graphite furnace atomic absorption spectrophotometry, after digestion with nitric acid and hydrogen peroxide. Erythrocytes account for about 66% of the manganese in whole blood, whereas the "buffy coat"--platelets and leukocytes--accounts for about 30%. The "buffy coat" components turn over more rapidly than do erythrocytes, so their manganese content may better indicate the body's manganese status.

Response of various indices of iron status to acute iron depletion produced in menstruating women by low iron intake and phlebotomy.

We investigated response sensitivities of indices of iron status to controlled iron depletion and repletion in 11 premenopausal women. The women were depleted of storage iron (as reflected by serum ferritin) through a combination of a low-iron diet and phlebotomy. They then consumed a diet containing 13.7 mg of iron per 2000 kcal, supplemented with either ascorbic acid or placebo (for 5 1/2 weeks) and a daily 50-mg iron supplement (for the subsequent 17 days). The relative sensitivities of different indices for detecting iron depletion were as follows: ferritin greater than % transferrin saturation greater than plasma iron greater than hemoglobin greater than hematocrit greater than zinc protoporphyrin (ZnPP) and erythrocyte protoporphyrin (EP). Ascorbic acid treatment during repletion, before iron supplementation, significantly (P less than 0.05) affected changes in hemoglobin, ZnPP, ZnPP/heme, and EP/heme. Changes in heme synthesis evidently do not occur until iron stores are depleted and, conversely, during iron repletion hematopoiesis must be satisfied before iron stores, as reflected by ferritin, increase. Thus, the use of only one index of iron status is of limited value for detecting iron depletion.

Short-term and long-term variability of indices related to nutritional status. I: Ca, Cu, Fe, Mg, and Zn.

Five free-living women (ages 28-38 y) and five women (ages 23-44 y) residing in a metabolic unit and eating a constant diet were assessed for variation in indices related to mineral nutrition. Blood was sampled once a month for five months, once a week for five weeks, and once a day for five days to assess analytical and biological variability. Analytical variability was determined by using concurrently run duplicate control samples prepared from plasma or serum pools. Of the measured indices, serum ferritin varied most, with intra-individual variance of 4.72% to 18.0%. Much of this variance may have been because of changes in iron status or in the analytical technique used. Intra-individual month-to-month variance for other indices ranged from 17% for superoxide dismutase (EC 1.15.1.1) to 1.5% for calcium. Correction for long-term analytical variation indicated that most of the variance was associated with the biological component. The higher biological variabilities of some indices, including ferritin or superoxide dismutase, need to be considered when nutritional status is being evaluated or when serial observations are made over a protracted period in clinical studies or trials.

Effects of dietary copper on human autonomic cardiovascular function.

Heart rate and blood pressure responses during supine rest, orthostasis, and sustained handgrip exercise at 30% maximal voluntary contraction were determined in eight healthy women aged 18-36 years who consumed diets varying in copper and ascorbic acid content. Copper retention and plasma copper concentration were not affected by diet. Enzymatic, but not immunoreactive, ceruloplasmin was lower (p less than 0.05) after the low copper and high ascorbic acid diet periods. Diet had no effect on resting supine heart rates, orthostatic responses in heart rate and blood pressure, or standing resting blood pressure. Systolic and diastolic blood pressures were increased significantly (p less than 0.05) during the handgrip test at the end of the low copper and ascorbic acid supplementation periods. Also, the ratio of enzymatic to immunoreactive ceruloplasmin decreased significantly during these dietary treatments. The mean arterial blood pressure at the end of the handgrip test was negatively (p less than 0.0004) correlated with the ceruloplasmin ratios. These findings indicate a functional alteration in human blood pressure regulation during mild copper depletion.