Molybdenum intake influences molybdenum kinetics in men.

The objectives of this study were to determine physiologic adaptations that occur when humans are exposed to a wide range of molybdenum intake levels and to identify the pathways that are influenced by dietary intake. Four males consumed each of 5 daily molybdenum intakes of 22, 72, 121, 467, and 1490 microg/d (0.23, 0.75, 1.3, 4.9, and 15.5 micromol/d) for 24 d each. During each treatment period, oral and intravenous doses of (100)Mo and (97)Mo were administered. Serial plasma, urine, and fecal samples were analyzed for labeled and unlabeled molybdenum. Compartmental modeling was used to determine rates of distribution and elimination at each dietary intake level. Three pathways were sensitive to daily molybdenum intake. With increasing intake, absorption and urinary molybdenum excretion increased, whereas the fraction deposited in tissues decreased. Kinetic analysis suggested a daily intake of 115-120 microg/d (1.20-1.25 micromol/d) would maintain initial plasma molybdenum levels at their prestudy values and that their prestudy dietary intake was well above the Recommended Dietary Allowance of 45 microg/d. The physiological adaptations to changing intake that the model demonstrated may help prevent molybdenum deficiency and toxicity.

Molybdenum kinetics in men differ during molybdenum depletion and repletion.

In this study we developed an expanded compartmental model of molybdenum (Mo) kinetics to determine rates of molybdenum distribution during molybdenum depletion and repletion. The model was based on a clinical study in which 4 men consumed a low-molybdenum diet of 22 microg/d (0.23 micromol/d) for 102 d, followed by a high molybdenum diet of 467 microg/d (4.9 micromol/d) for 18 d. Stable isotopes 100Mo and 97Mo were administered orally and intravenously, respectively, at several time points during the study, and serial samples of plasma, urine, and feces were analyzed for 100Mo, 97Mo, and total Mo. Based on plasma, urine, and fecal molybdenum levels, kinetic parameters of distribution and elimination were determined. The rates of molybdenum distribution and elimination were different during depletion and repletion. During high intake, urinary molybdenum excretion was greater than during low intake. In addition, fractional tissue storage of molybdenum was lower during high intake than during low intake. This suggests that low intake results in an adaptation to conserve body Mo, and that high intake results in an adaptation to eliminate Mo. The model also suggested that food-bound molybdenum was approximately 16% less bioavailable than purified Mo. Finally, under the conditions of this study, the model suggested that an intake of 43 microg/d (0.45 micromol/d) would be sufficient to maintain plasma molybdenum levels at steady state. This is a minimum estimate because subjects in this study were in a molybdenum-sparing state. These findings provide an understanding of the adaptations in molybdenum metabolism that take place during depletion and repletion.

Long-term high copper intake: effects on copper absorption, retention, and homeostasis in men.

BACKGROUND: Numerous studies have examined the effect of low and adequate intakes of copper on absorption and retention, but little information is available on the regulation of absorption and retention of copper when intake is high. OBJECTIVE: A study was conducted in men to determine the effect of long-term high copper intake on copper absorption, retention, and homeostasis. DESIGN: Nine men were confined to a metabolic research unit (MRU) for 18 d and were fed a 3-d rotating menu containing an average of 1.6 mg Cu/d. They continued the study under free-living conditions for 129 d, supplementing their usual diets with 7 mg Cu/d. They then returned to the MRU for 18 d and consumed the same diet as during the first period, except that copper intake was 7.8 mg/d. The stable isotope (63)Cu was fed to 3 subjects and infused into the other 6 on day 7 of each MRU period, and complete urine and stool collections were made throughout the study. Total copper and (63)Cu were determined by inductively coupled plasma mass spectrometry. Copper absorption, excretion, and retention were calculated on the basis of dietary, urinary, and fecal copper and (63)Cu. RESULTS: Results were as follows when comparing the high copper intake with the usual intake: fractional copper absorption was significantly lower, but the amount absorbed was significantly higher; excretion of the infused (63)Cu was significantly faster; and total retention was significantly higher. CONCLUSIONS: Homeostatic regulation of copper absorption and retention helped to minimize the amount of copper retained with high copper intake but was not sufficient to prevent retention of >0.6 mg Cu/d.

Effect of long-term, high-copper intake on the concentrations of plasma homocysteine and B vitamins in young men.

OBJECTIVE: We evaluated the effect of a long-term, high-copper intake on plasma total homocysteine, folate, vitamin B12, and pyridoxal-5'-phosphate concentrations in humans. METHODS: Nine healthy young men were confined in a metabolic research unit for 18 d and fed 3-d rotation diets supplying an average of 1.6 mg of copper per day followed by 129 d of free-living conditions when they received 7 mg of copper per day in addition to their usual diets. The subjects returned to the metabolic research unit for the second 18-d period and were given the same diets as during the first 18 d with the exception that the copper intake was 7.8 mg/d. There was no apparent biochemical indication that the subjects were deficient in copper before the large-dose copper intake. Blood samples were obtained at the end of the first and second 18-d periods at the metabolic research unit, and plasma concentrations of total homocysteine, folate, vitamin B12, and pyridoxal-5'-phosphate were measured. RESULTS: The long-term, high-copper intake resulted in small but significant decreases in plasma concentrations of total homocysteine and folate. There was no effect of the high-copper intake on plasma concentrations of vitamin B12 and pyridoxal-5'-phosphate. CONCLUSIONS: These findings can be explained by our previous observation in rats suggesting that methionine synthase is copper dependent and that the metabolism of homocysteine and folate is regulated in part by copper nutriture. It may be necessary to consider copper nutriture for the interpretation of plasma concentrations of total homocysteine in humans.

Kinetic parameters and plasma zinc concentration correlate well with net loss and gain of zinc from men.

The search for a reliable, convenient indicator of Zn status was the focus of research for several decades. Plasma Zn concentration is still the most widely used clinical measurement, despite the known problems of interpretation. More recently, researchers suggested that isotopically determined kinetic parameters, such as the exchangeable Zn pool (EZP), may more accurately and reliably reflect body Zn status. The objective of this study was to examine the relationship between net body Zn loss and gain during acute changes in dietary Zn intake with biochemical and kinetic indices of Zn status. Five men participated in an 85-d Zn depletion/repletion study. Net body Zn loss and gain were determined from the difference between dietary plus intravenously administered Zn and Zn excretion. Biochemical indicators of Zn status included plasma Zn, plasma alkaline phosphatase activity, and plasma retinol binding protein concentration. Following intravenous administration of (70)Zn or (67)Zn, a compartmental model was used to determine EZP mass, fractional Zn absorption, endogenous zinc excretion (EZE), and plasma Zn flux. The changes in total body zinc correlated best with changes in plasma Zn (r(2) = 0.826, P < 0.001), EZE (r(2) = 0.773, P < 0.001), and plasma Zn flux (r(2) = 0.766, P < 0.001). This study confirms that plasma Zn concentration is a valid indicator of whole-body Zn status in the absence of confounding factors; however, further research is needed to determine how kinetic parameters respond to conditions where plasma Zn concentration is known to be unreliable.

Long-term high copper intake: effects on indexes of copper status, antioxidant status, and immune function in young men.

BACKGROUND: Short-term high copper intake does not appear to affect indexes of copper status or functions related to copper status, but the effects of long-term high copper intake are unknown. OBJECTIVE: A study was conducted in men to determine the effect of long-term high copper intake on indexes of copper status, oxidant damage, and immune function. DESIGN: Nine men were confined to a metabolic research unit (MRU) for 18 d and were fed a 3-d rotating menu providing an average of 1.6 mg Cu/d. The men continued the study under free-living conditions for 129 d and supplemented their usual diets with 7 mg Cu/d. The men then returned to the MRU for 18 d of the same diet as during the first period, except that copper intake was 7.8 mg/d. Plasma copper, ceruloplasmin activity, ceruloplasmin protein, plasma malondialdehyde, benzylamine oxidase activity, erythrocyte superoxide dismutase, hair copper, urinary copper, and urinary thiobarbituric acid-reactive substances were measured during each MRU period. RESULTS: Ceruloplasmin activity, benzylamine oxidase, and superoxide dismutase were significantly higher at the end of the second MRU period than at the end of the first. Urinary copper excretion, hair copper concentrations, and urinary thiobarbituric acid-reactive substances were significantly higher during the second MRU period than during the first. Polymorphonuclear cell count, the percentage of white blood cells, lymphocyte count, and interleukin 2R were affected by copper supplementation. Antibody titer for the Beijing strain of influenza virus was significantly lower in supplemented subjects after immunization than in unsupplemented control subjects. CONCLUSIONS: Under highly controlled conditions, long-term high copper intake results in increases in some indexes of copper status, alters an index of oxidant stress, and affects several indexes of immune function. The physiologic implications of these changes are unknown.

Plasma molybdenum reflects dietary molybdenum intake.

The relationship between plasma molybdenum (Mo) and dietary intake has not been investigated in humans. We developed an isotope dilution method to determine molybdenum in 0.5 mL blood plasma by ICP-MS and conducted a study to determine the effect of dietary intake on plasma molybdenum. Twelve young men consumed a very low Mo diet (22 microg/day) for 24 days while confined to the WHNRC metabolic research unit and plasma molybdenum was monitored. (97)Mo was infused in four of the subjects (Group 1) to follow its clearance from the blood. The other eight remained in unit for 120 days (an additional 96 days). Four consumed the 22 microg/day molybdenum diet for 102 days followed by 467 microg/day for 18 days (Group 2). and four consumed five levels of dietary molybdenum for 24 days each (Group 3). (100)Mo was added to the diet one or more times at each dietary level. Total plasma molybdenum and (100)Mo were monitored throughout the study. Plasma molybdenum in the 12 subjects decreased from 8.2 +/- 0.5 to 6.1 +/- 0.5 nmol/L after 13 days of low molybdenum intake and was 5.1 +/- 0.5 nmol/L after 24 days. In Group 2, average plasma molybdenum was 7.8 +/- 0.9 nmol/L at the beginning of the study, 5.4 +/- 0.4 nmol/L during the 102 days low molybdenum period, and 16.5 +/- 0.6 nmol/L during the high molybdenum period. Plasma molybdenum in Group 3 was 4.2 +/- 2.1 nmol/L at 22 microg/day; 5.8 +/- 2.5 nmol/L at 72 microg/day; 6.6 +/- 2.3 nmol/L at 121 microg/day; 19.7 nmol/L +/-2.1 at 467 microg/day; and 43.9 +/- 2.1 nmol/L at 1490 microg/day. The results demonstrate that, in contrast to most other essential minerals, plasma molybdenum reflects low and high dietary molybdenum intakes within 14 days and may a useful indicator of low and high dietary intakes.

A compartmental model of magnesium metabolism in healthy men based on two stable isotope tracers.

The aim of this study was to build a compartmental model of magnesium (Mg) kinetics by using data collected from six healthy adult men after oral administration of 26Mg and intravenous administration of 25Mg. Blood, urine, and feces were collected for 12 days after administration of the isotopes. Isotopic ratios were determined by inductively coupled plasma-mass spectrometry. Data were analyzed for each subject using SAAMII. We began with a compartmental model previously proposed (Avioli LV and Berman M. J Appl Physiol 21: 1688-1694, 1966) and developed an alternative approach to resolve the discrepancy between model-predicted curves and experimental data. This analysis enables the exploration of 25% of total body Mg that exchanges rapidly from plasma compartment with two extraplasma pools. One of the extraplasma compartments contains 80% of the exchangeable Mg with a transport rate of 48 +/- 13 mg/h. The second exchanges 179 +/- 88 mg of Mg/h. The model permitted estimation of kinetic parameters as well as fractional Mg absorption and fecal endogenous excretion.

Comparison of stable-isotope-tracer methods for the determination of magnesium absorption in humans.

BACKGROUND: The double-labeling (DL) method for determining magnesium absorption is less cumbersome than is the fecal monitoring method, which has been used most often, but it has not been validated. OBJECTIVE: The aim of this study was to compare methods and several sampling protocols for determining magnesium absorption to establish a simple and reliable alternative to the fecal monitoring approach. Fecal monitoring was used as the standard against which the DL methods based on urine data (DLU), plasma data (DLP), and plasma kinetics with the use of a deconvolution analysis (DP) were compared. DESIGN: Six healthy adult men received 70 mg (26)Mg orally and 30 mg (25)Mg intravenously. Multiple blood samples and complete urine and fecal samples were collected over 12 d. Stable-isotope ratios were determined by inductively coupled plasma mass spectrometry. RESULTS: Results from DLU were not significantly different from the fecal monitoring reference value (0.48 +/- 0.05; +/- SD) when based on 3-d urine pools from 72 to 144 h (0.54 +/- 0.04) and when based on the 24-h urine pools from 48 to 72 h (0.49 +/- 0.06), 72 to 96 h (0.51 +/- 0.11), and 96 to 120 h (0.50 +/- 0.06). Results with the DLP method 72 h after isotope administration also compared well with those with the fecal monitoring method (0.54 +/- 0.09). Magnesium absorption was 0.47 +/- 0.06 with the DP method, which also agreed with the fecal monitoring value. CONCLUSIONS: The DL methods are an alternative to fecal monitoring when applied within the appropriate time intervals. Therefore, DLU-the simplest and least invasive approach-is recommended for determining magnesium absorption.

Isotope ratios of trace elements in samples from human nutrition studies determined by TIMS and ICP-MS: precision and accuracy compared.

Stable isotopes are used with increasing frequency to trace the metabolic fate of minerals in human nutrition studies. The precision of the analytical methods used must be sufficient to permit reliable measurement of low enrichments and the accuracy should permit comparisons between studies. Two methods most frequently used today are thermal ionization mass spectrometry (TIMS) and inductively coupled plasma mass spectrometry (ICP-MS). This study was conducted to compare the two methods. Multiple natural samples of copper, zinc, molybdenum, and magnesium were analyzed by both methods to compare their internal and external precision. Samples with a range of isotopic enrichments that were collected from human studies or prepared from standards were analyzed to compare their accuracy. TIMS was more precise and accurate than ICP-MS. However, the cost, ease, and speed of analysis were better for ICP-MS. Therefore, for most purposes, ICP-MS is the method of choice, but when the highest degrees of precision and accuracy are required and when enrichments are very low, TIMS is the method of choice.

Stable isotope techniques in human nutrition research: concerted action is needed.

Use of stable isotopes in applied nutrition has been a developing field for over 60 years. The past 20 years has seen improvements in computer and mass-spectrometer technology that has opened up even greater possibilities in the understanding of human metabolism. While improvements in technology can bring great opportunities, it can also cause problems if there is no consensus among the stable isotope user-community on standardization of new techniques and methods. Users of stable isotopes have traditionally been split into two groups; those who work with heavy isotopes (e.g., 58Fe, 70Zn) and those who work with light isotopes (e.g., 2H218O). Standardization issues have been addressed by the light isotope users and awareness of this type of problem is starting to emerge within the heavy isotope community.