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Unequal distribution of a stable isotopic calcium tracer between casein and whey fractions of infant formulas, human milk and cow's milk.

Measurement of calcium absorption with tracers assumes a complete equilibration of tracer with milk calcium. In this study, the equilibration of tracer between the micellar casein and soluble fractions (primarily whey) of infant formulas, human milk and cow's milk was measured in vitro using milk samples enriched with 42Ca and analyzed by thermal ionization mass spectrometry. Incomplete equilibration of tracer occurred with the micellar casein fraction of all milks. The least equilibration with micellar casein was found with premature infant formula, for which the ratio of slopes of the equilibration lines (whey/casein) was 8.5/1. These differences may be due to Ca-casein binding in cow's milk-based formulas. The effects of the lack of tracer equilibration in vivo cannot be determined. However, unequal bioavailability of casein- vs. whey-bound Ca may exist.

Absorption

Calcium absorption in children estimated from single and double stable calcium isotope techniques.

In order to determine whether the specific activity in a single serum sample estimates calcium absorption, six healthy children participated in a study using stable isotopic calcium tracers, one given orally and the second intravenously. High-resolution, fast atom bombardment mass spectrometry was used to quantify the 44Ca and 42Ca tracers in serum and urine. Subjects ingested 250 mg of calcium (215 mg calcium enriched with 35 mg 44Ca) in the form of a chewable calcium citrate malate tablet with a standard meal, followed 30 min later with an i.v. injection of 42Ca tracer. Blood for tracer determinations was obtained at 90, 120, 150, 180, and 300 min after oral ingestion, and a urine sample was obtained 24 h after oral calcium tracer administration. The average calcium absorption estimated from the ratio of urinary tracers was 41.4 +/- 8.2%. This study indicates that the level of oral tracer in serum taken 150 min post-ingestion is significantly correlated (r = 0.85, p less than 0.05) with calcium absorption, as determined by the tracer levels in the urine. These results show that an oral stable isotopic tracer coupled with a single blood sample can be used to estimate calcium absorption in children.

Absorption

Calcium metabolism in cancer. Studies using calcium isotopes and immunoassays for parathyroid hormone and calcitonin.

Studies of calcium metabolism in 38 patients with cancer indicated that: 1) intestinal absorption of calcium was reduced in patients with skeletal metastases and in those with hypercalcemia; 2) calcium-47 space (a measurement of bone turnover rate) was high in the patients with skeletal metastases; 3) hypercalcemic patients had higher urinary and endogenous fecal excretion of calcium than those who were normocalcemic; 4) levels of plasma immunoreactive parathyroid hormone were similar in normo- and hypercalcemic patients, but the levels for a given serum calcium in malignant disease were lower than those in primary hyperparathyroidism; and 5) some patients had elevated calcitonin levels. Hypercalcemia complicating malignant disease is therefore not due to hyperabsorption or diminished excretion of calcium, and a low calcium diet is unlikely to benefit these patients. Measurement of 47Ca space could be of use in monitoring therapy of patients with skeletal metastases, and measurement of plasma parathyroid hormone could be useful in the differential diagnosis of hypercalcemia.

Adult

Direct measurement of dietary fractional absorption using calcium isotopic tracers.

Fractional dietary Ca absorption, 'a', is measured by determining the ratio of two stable isotopic tracers, one of them orally (44Ca + 0.2-0.5 mg/kg) and the other intravenously (42Ca / 0.02-0.1 mg/kg). Thermal ionization mass spectometry (TIMS) is used to measure the perturbation of natural abundance isotope ratios (delta % excess). Typical sensitivity of the TIMS permits detection of a 2.5 delta % excess change from the natural Ca isotope ratio with relative standard deviations of about 0.5%. At sufficiently long times absorption becomes constant so that 'a' is determined by a product of constants and a measured ratio.

Adolescent

Classification of idiopathic hypercalciuric patients by isotopic calcium absorption: a comparison with oral calcium tolerance test.

To test the accuracy of calcium tolerance test in estimating calcium absorption, we have measured the radioactive calcium absorption (expressed as Fx) in 27 patients with IH and renal calcium stones. The results of this test were compared with those of a standard oral calcium tolerance test. Although only seven of nine AH patients displayed normal fasting calcium excretion, they all displayed Fx values above normal and a normal parathyroid activity. Conversely, only 5 of our 18 RH patients demonstrated a hyperabsorption of radioactive calcium and an elevation in iPTH and cAMP above normal limits, yet all of them showed an increased calciuric response to an oral calcium challenge. Calcium absorption was inversely related to iPTH (r = -082; P less than 0.001) and cAMP (r = -064 P less than 0.05) in AH, but directly proportional to these parameters (r = 0.62 P less than 0.001 and r = 0.46 P less than 0.05, respectively) in RH patients. In view of these results, two ratios, iPTH/Fx and cAMP/Fx were used to discriminate between the two groups of patients. Both ratios were over normal limits in all RH patients and within normal range in all but one AH patient. Furthermore, no overlap was found between the two groups. Conversely, we were unable to completely separate AH from RH subjects on the basis of the oral calcium tolerance test, since in both groups the fasting and the absolute (or percentage) changes in urinary calcium, cAMP and blood iPTH levels following oral calcium loading, overlapped in each instance.(ABSTRACT TRUNCATED AT 250 WORDS)

Absorption

Absent intestinal response to calciferols in hereditary resistance to 1,25-dihydroxyvitamin D: documentation and effective therapy with high dose intravenous calcium infusions.

We describe a patient with an absent intestinal response to 1,25-dihydroxyvitamin D [1,25-(OH)2D] and the beneficial effects of treatment with high dose iv calcium infusion. The patient presented with severe rickets despite therapy with extraordinarily high doses of 1 alpha-hydroxyvitamin D3 or 1,25-(OH)2D3. Unidirectional intestinal fractional calcium absorption when he was not treated with any calciferol was 14% (normal, 20-70%), as measured with stable calcium isotopes; no increase in calcium absorption occurred when serum 1,25-(OH)2D levels were more than 50-fold elevated. Cultured skin fibroblasts contained no detectable 25-hydroxyvitamin D3-24-hydroxylase activity in response to 1,25-(OH)2D3 (10(-9)-10(-6) mol/L). High dose iv calcium infusions and oral phosphorus supplementation for 135 days improved or normalized biochemical parameters and resulted in radiographic healing of the rachitic lesions. We conclude that 1) this patient had no response to 1,25-(OH)2D3 in vivo and in vitro; 2) long term parenteral calcium infusions were effective therapy in managing the patient's severe resistance to 1,25-(OH)2D; and 3) stable calcium isotopes are useful for measuring low levels of fractional calcium absorption.

Alkaline Phosphatase

Calcium absorption in Crohn's disease.

Calcium absorption and endogenous loss of calcium were measured in a group of patients with Crohn's disease, using a simultaneous metabolic balance and calcium isotope regimen. Calcium malabsorption resulting in negative calcium balance was found in only 4 of 31 patients with Crohn's disease. No elevation of endogenous fecal calcium or total secreted intestinal calcium was observed in 10 patients studied, regardless of the level of net or true calcium absorption. Correlation between calcium balance and serum protein loss was observed, but no association was noted with intestinal fat excretion, d-xylose absorption, bacterial colonization of the jejunum, or glucocorticosteroid therapy. The results indicate that in this group of patients with Crohn's disease involving different areas of the intestine, calcium malabsorption occurred infrequently and that the levels of calcium excretion correlated best with enteric protein loss.

Adolescent

Chromaffin cell calcium channel kinetics measured isotopically through fast calcium, strontium, and barium fluxes.

Fast Ca2+ uptake into K+-depolarized cultured bovine adrenal chromaffin cells has been isotopically measured in a time scale of 1-10 s. Depolarized cells retained as much as 80-fold 45Ca2+ taken up by resting cells; Ca2+ was not taken up by fibroblasts or endothelial-like cells. Because Ca2+ entry was inhibited by inorganic (La3+, Co2+, Mg2+) and organic (nifedipine) Ca2+ channel antagonists and enhanced by the Ca2+ channel activator Bay-K-8644, it seems clear that Ca2+ gains access to the chromaffin cell cytosol mainly through specific voltage-dependent Ca2+ channels. Ca2+ uptake evoked by 59 mM K+ was linear during the first 5 s of stimulation and continued to rise at a much slower rate up to 60 s. The rate of Ca2+ entry became steeper as the external [Ca2+] increased; initial rates of Ca2+ uptake varied from 0.06 fmol/cells . s at 0.125 mM Ca2+ to 2.85 fmol/cell . s at 7.5 mM Ca2+. The early 90Sr2+ uptake was linear but faster than Ca2+ uptake and later on was also saturated; 133Ba2+ was taken up still at a much faster rate and was linear for the entire depolarization period (2-60 s). Increased [K+] gradually depolarized chromaffin cells; Ca2+ and Sr2+ uptakes were not apparent below 30 mM K+ but were linear for 30 to 60 mM K+. In contrast, substantial Ba2+ uptake was seen even in K+-free solutions; and in 5.9 mM K+, Ba2+ uptake was as high as Ca2+ uptake obtained in 60 mM K+. Five to ten-second pulses of 45Ca2+, 90Sr2+, or 133Ba2+ given at different times after pre-depolarization of chromaffin cells served to analyze the kinetics of inactivation of the rates of entry of each divalent cation. Inactivation of Ca2+ uptake was faster than Sr2+, and Ba2+ uptake inactivated very little. Neither voltage changes nor Ca2+ ions passing through the channels seems to cause their inactivation; however, experiments aimed to manipulate the levels of internal Ca2+ using the cell-permeable chelator Quin-2 or the ionophore A23187 strongly suggest that intracellular Ca2+ levels determine the rates of inactivation of these channels.

Adrenal Gland Neoplasms

Recent studies of human calcium metabolism using stable isotopic tracers.

Stable isotopes of calcium are used safely as tracers for calcium in human populations ranging in age from infants to postmenopausal women. Thermal ionization mass spectrometry is used to measure calcium isotope ratios with relative accuracies of about 1% for natural abundance ratios at precisions of about 1% relative to the mean. Perturbations of natural abundance ratios are determined for the calcium in blood, urine, and feces with a limit of detection of about 2 delta % excess. The mathematical rationale for clinical studies of fractional absorption of dietary calcium and the kinetics of calcium's internal distribution are presented.

Animals

A stable tracer technique for calcium metabolism studies.

A tracer technique using the stable isotope calcium-48 was developed to facilitate studies of human calcium metabolism. Small samples of 1 ml serum were used. Calcium levels were determined with good precision using neutron-activation analysis after pre-concentration of the serum calcium by oxalate precipitation. The method has sufficient sensitivity to detect induced dilutions of the tracer by administering other less costly isotopes of calcium. The ability to utilize small volumes of sample is of direct benefit to paediatric metabolism research. The favourable results obtained for two calcium loading tests demonstrate the applicability of the technique to intestinal absorption studies.

Calcium