A multi-center comparison of dual energy X-ray absorptiometers: in vivo and in vitro soft tissue measurement.

OBJECTIVE: To assess intra- and inter-site soft tissue variability by dual energy X-ray absorptiometry (DXA). DESIGN: Cross-sectional trial. SETTING: Three medical research institutions. SUBJECTS: Five humans (in vivo) and four phantoms (in vitro), configured from two whole body phantoms with artificial skeletons and thickness overlays. INTERVENTIONS: Duplicate total-body DXA scans were performed on all subjects at each institution within a 15 d period. RESULTS: All intra-site coefficients of variation (CV) were < 0.5% for total tissue mass, but in vitro and in vivo Cvs were 7.2% and 2.3% for fat mass (FM) and 2.5% and 0.9% for lean mass (LM), respectively. Several total-body and regional FM and LM measurements were significantly different between sites (P < 0.05), with percent differences between sites ranging from 2.6-13.3% for FM and from 1.6-13.6% for LM. Site 2 was consistently lower for FM and Site 3 was consistently lower for LM. CONCLUSIONS: These results stress the need for both rigorous and standardized cross-calibration procedures for soft tissue measurement by DXA.

A multicenter comparison of dual-energy X-ray absorptiometers: in vivo and in vitro measurements of bone mineral content and density.

As part of a multicenter research project, we compared dual-energy X-ray absorptiometry (DXA) instruments at three research centers (sites 1, 2 and 3) to determine both intra- and intersite variability of bone mineral content (BMC), bone mineral density (BMD), and bone area (BA). Scans of the total body and lumbar spine were performed in duplicate on five humans (in vivo), and scans of the total body were performed on two whole body phantoms with artificial skeletons and thickness overlays (in vitro) at all sites over 15 days. The average intrasite variability in two consecutive total body BMD measurements, expressed as a percent difference, was significantly higher in vitro, 1.74 +/- 1.97%, than in vivo, 0.71 +/- 0.38% (p < 0.05). Average intrasite variability of the in vivo lumbar spine BMD was 1.08 +/- 1.12%. The intersite coefficients of variation for all BMD, BMC, and BA measurements were < 2.0%. The total body BMD from site 2 was systematically lower than at sites 1 and 3 both in vivo and in vitro (p < 0.05) with no differences in BMC and BA. Although significant, the total body BMD differences between sites were small (< 1.2%) in vivo compared with in vitro (< 2.6%) and are encouraging for the comparison and pooling of human data from multicenter trials, provided that appropriate standardized cross-calibration and analysis procedures are applied.

Calibration of the delayed-gamma neutron activation facility.

The delayed-gamma neutron activation facility at Brookhaven National Laboratory was originally calibrated using an anthropomorphic hollow phantom filled with solutions containing predetermined amounts of Ca. However, 99% of the total Ca in the human body is not homogeneously distributed but contained within the skeleton. Recently, an artificial skeleton was designed, constructed, and placed in a bottle phantom to better represent the Ca distribution in the human body. Neutron activation measurements of an anthropomorphic and a bottle (with no skeleton) phantom demonstrate that the difference in size and shape between the two phantoms changes the total body calcium results by less than 1%. To test the artificial skeleton, two small polyethylene jerry-can phantoms were made, one with a femur from a cadaver and one with an artificial bone in exactly the same geometry. The femur was ashed following the neutron activation measurements for chemical analysis of Ca. Results indicate that the artificial bone closely simulates the real bone in neutron activation analysis and provides accurate calibration for Ca measurements. Therefore, the calibration of the delayed-gamma neutron activation system is now based on the new bottle phantom containing an artificial skeleton. This change has improved the accuracy of measurement for total body calcium. Also, the simple geometry of this phantom and the artificial skeleton allows us to simulate the neutron activation process using a Monte Carlo code, which enables us to calibrate the system for human subjects larger and smaller than the phantoms used as standards.

Racial differences in femoral dimensions and their relation to hip fracture.

White women have a higher rate of age-specific hip fractures than black women. Recently, femoral dimensions have been implicated in osteoporotic fractures. To study racial differences in femoral dimensions, dual X-ray absorptiometry scans were obtained for two similar groups of 50 white women and 50 black women. We measured the hip axis length (the distance from below the lateral aspect of the greater trochanter to the inner pelvic brim), the neck width and the neck/shaft angle on the scan print-out. The observer was masked to the race of the subject. The results were analyzed using the independent t-test and showed that the hip axis length and the neck width were significantly longer in the white women than in the black women (p values < 0.05 and < 0.02 respectively) but that the neck/shaft angle was not statistically different in the two groups. We conclude that femoral geometry differs among races. Whether this contributes to the lower risk of hip fracture in black women will require prospectively based studies.

Relationship of menopause to skeletal and muscle mass.

Cross-sectional and longitudinal changes in body composition with age were examined in white women to determine the relationship of body cell mass to menopause and of body fat to bone mass. There was statistical evidence for a curvilinear component to loss of total body potassium with negligible rates of loss before menopause. Longitudinal measurements also indicated a relationship between the proximity to menopause and the rate of loss of potassium. Total body potassium was significantly related to total body calcium and bone density of the spine, radius, and femoral neck. Total body fat was not related to any of these measurements. We found no evidence that adiposity plays a major role in protecting against bone loss.

Premenopausal bone mass is related to physical activity.

The relationship between physical activity and bone mass was investigated in 24 healthy, white, premenopausal women (mean age [+/- SE], 39.0 +/- 1.39 years). Physical activity was determined by a sensor that measures movement of the trunk, and bone mineral levels were determined by means of single- and dual-photon absorptiometry and neutron activation analysis. Total physical activity levels were related both to bone mineral density of the spine (r = .41) and to total body calcium levels (r = .51). There was no significant relationship between the bone density of the distal portion of the radius and activity (r = .20). Nonparametric analysis and stepwise multiple regression analysis revealed negative correlations between cigarette smoking and bone density of the spine and radius. These data suggest that the level of physical activity in sedentary white women may be a determinant of peak total skeletal mass and bone density of the spine.

Serum osteocalcin and total body calcium in normal pre- and postmenopausal women and postmenopausal osteoporotic patients.

Serum osteocalcin was measured in 51 normal pre- and 114 postmenopausal women and in 41 postmenopausal osteoporotic patients. Total body calcium (TBCa) was determined in the same individuals by neutron activation analysis. Many of the perimenopausal nonosteoporotic women had increased serum osteocalcin values, but 15 yr or more after the menopause most of the women had serum osteocalcin levels in the normal range. Comparing normal women before and after menopause, the mean serum osteocalcin levels [7.8 +/- 4.7 (+/- SE) and 10.1 +/- 9.4 ng/mL] were not significantly different; however, the TBCa values (898 +/- 99 and 806 +/- 111 g) were significantly different (P less than 0.001). When the normal postmenopausal women were regrouped according to high vs. low osteocalcin values, TBCa and phosphorus content as well as forearm linear bone density were significantly lower in the high osteocalcin group, even though most of the other variables, including urinary hydroxyproline excretion, serum alkaline phosphatase, age, height, and weight, were not different. Osteoporotic women had a mean serum osteocalcin concentration of 17.4 +/- 8.6 ng/ml and a TBCa of 657 +/- 83 g, both significantly different from the respective values in normal and pre- and postmenopausal women (P less than 0.001 for both variables in comparison to each group). These data suggest that high serum osteocalcin levels, at least on a group basis, are an index of low skeletal mass.

Effects of elevated lead and cadmium burdens on renal function and calcium metabolism.

To assess the pathophysiologic significance of increased body burdens of lead and cadmium, detailed renal function studies and evaluation of calcium, phosphorus, and vitamin D metabolism were carried out in 38 industrial workers exposed to lead and cadmium for 11 to 37 yr. Body burden of lead, as assessed by x-ray fluorescence measurement of tibia lead content, was elevated in 58% of the men and, when assessed by excretion of lead after Ca-EDTA infusion, was elevated in 36%. Liver or kidney cadmium burden, as assessed by neutron activation analysis, was elevated in 31%. Creatinine clearance was normal in all workers. One worker was hyperuricemic and two were proteinuric; three had increased beta 2 microglobulin excretion and one had diminished urinary acidifying ability. Maximal urinary concentrating ability was abnormal in a significant fraction, i.e., 52% of the men. Individuals with a high lead burden had a slight decrease in mean serum phosphorus but no accompanying phosphaturia. There was no abnormality of serum calcium. Twenty-two percent of subjects were hypercalciuric and two had low vitamin D levels, but these abnormalities bore no relation to heavy metal burden. In this carefully characterized group of men with chronic lead and calcium exposure, definite, if subclinical, effects on renal function and serum phosphorus but not calcium or vitamin D metabolism were demonstrable.

Effect of hypophysectomy on the occupied and unoccupied binding sites for 1,25-dihydroxyvitamin D3 in rat intestine.

It is not known why the intestinal active transport of calcium per unit of mucosal mass is not affected by hypophysectomy (HX) even though serum 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] and intestinal calcium-binding protein are decreased. In order to study the effect of HX on the quantity of intestinal receptor of 1,25(OH)2D3 and its binding characteristics, the intestinal total occupied and unoccupied binding sites for 1,25(OH)2D3 were measured, by the use of the mercurial reagent mersalyl, in the intestine of HX and age-matched control rats. In addition, the effect of bovine growth hormone (bGH) replacement on the quantity of both binding states was examined in the HX rats. Results of Scatchard analysis and sucrose density gradients showed that the 3.5S receptor of the HX rat intestine was not distinguishable from that seen in the intact rat intestinal cytosol. Under vitamin D-supplemented conditions, HX was shown to reduce the levels of occupied receptors when the data were expressed on the basis of cytosolic protein. The reduced occupied sites could, in fact, have resulted from the reduction in plasma 1,25(OH)2D3 levels. No synthesis rates were determined. The unoccupied and total binding sites for 1,25(OH)2D3 per length of intestine were lower in the HX group than in the intact group. Administration of bGH resulted in an increase of endogenously occupied binding sites without affecting the total binding activity. Under vitamin D-depleted (-D) conditions, the total binding activity (intestinal) for 1,25(OH)2D3 was increased in the intact but not in the -DHX rats. Administration of bGH to the -DHX rat resulted in no effect on the binding levels of 1,25(OH)2D3 receptor.(ABSTRACT TRUNCATED AT 250 WORDS)

Women at risk for developing osteoporosis: determination by total body neutron activation analysis and photon absorptiometry.

With stepwise multiple logistic regression (MLR), probabilistic classification equations were developed to identify asymptomatic women who are at risk for development of fracture of the spine. Clinically normal women with low TBCa/square root H ratios can be classified as at risk for osteoporosis prior to their developing spinal compression fractures. With receiver operating characteristic (ROC) analysis, it was possible to verify the accuracy of the MLR model to discriminate "normal" women at risk, with high sensitivity and specificity. With the MLR model, discrimination of osteoporotic women (50-59 years) was made correctly for 86.2% of the total osteoporotic subjects with the TBCa data. Similar models were derived from the photon absorptiometry data. From the spinal density (BDs) data, correct classification in the 50-59 year group was 55.6% of the total osteoporosis subjects; from the radius density (BMCr) data, the corresponding value was 31%. The highest probability of identifying osteoporosis in all age categories was, therefore, on the basis of TBCa data. Similar, but less accurate discrimination was achieved with the BDs and BMCr data. These conclusions were confirmed by the application of receiver operating characteristic (ROC) analysis. Correct identification of the population at risk permits the timely and efficient application of therapeutic programs prior to onset of fracture. In a serial study of 104 peri-menopausal women, for example, it was possible to determine the P value for individuals measured annually over a 3-10 year period and thus to predict normal individuals at risk for developing osteoporosis each year.

In vivo bone lead measurements: a rapid monitoring method for cumulative lead exposure.

Lead concentrations (microgram/g wet weight) in human bone (tibia) were measured noninvasively in vivo employing an X-ray fluorescence technique. Forty-five workers who had been subjected to chronic industrial exposure were found to have a mean bone lead content of 52.9 micrograms/g wet weight (0 to 198 micrograms/g). In addition to bone lead content, blood lead, body burden of lead as assessed by urinary lead excretion after EDTA chelation, zinc protoporphyrin, and unstimulated urinary lead excretion were evaluated. The results suggest that the in vivo measurement of tibia lead content may serve as an acceptable indicator of body lead burden and provide a practical technique for lead screening purposes. The correlation coefficient between X-ray fluorescence findings and lead excretion following Ca-EDTA administration is 0.69; p less than 0.001.

Nuclear resonance scattering measurement of human iron stores.

Hepatic iron stores were measured noninvasively in 31 patients (thalassemia, hemodialysis, hemosiderosis, refractory anemia) with suspected iron overload, employing a nuclear resonance scattering (NRS) technique. The thalassemia patients were undergoing desferrioxamine chelation therapy during the NRS measurements. The hemodialysis patients were measured before chelation therapy. Iron levels measured by NRS were in general agreement with those determined in liver biopsies by atomic absorption spectroscopy. In addition, NRS measurements from the thorax of some of these patients suggest that this method may also prove useful for clinical assessment of cardiac iron.

Assessment of cellular mass and lean body mass by noninvasive nuclear techniques.

With the nuclear techniques employed in this study, it has been possible to measure total body potassium (TBK), lean body mass (LBM), and body cell mass (BCM). The latter two are derived independently of TBK. LBM is derived from the sum of total body water (TBW), total body protein, and bone mineral ash measurements. The BCM value is, in turn, based on the difference between the LBM (as measured above) and the sum of the extracellular water and extracellular solids. It was demonstrated in 123 normal individuals that, although the TBK/LBM ratio decreases with age, the TBK/BCM ratio does not change significantly with age. It can be deduced that the ratio of TBK to intracellular water should be relatively constant with age. This relative constancy with age was demonstrated independently with the absolute measurements of TBK and intracellular water in normal individuals. Because the BCM is physiologically and chemically more homogeneous than LBM and because it reflects the actively metabolizing cellular compartment more accurately than LBM, it is the preferred parameter to be used for reference or normalization in body composition studies. For most applications, BCM is readily derived from TBK measurement by either whole body counting or isotope dilution techniques.

Effect of weight reduction on the renin-aldosterone axis.

Eight normotensive obese subjects participated in an inpatient study designed to determine the effect of a constant sodium intake (150 mEq) on the renin-aldosterone axis during 12 weeks of weight reduction. Two 800-calorie (3,200 kj) ketogenic diets, differing in carbohydrate content (10 g vs 70 g) were used for the study. Supine and upright plasma renin activity (PRA) and serum aldosterone (SA) were determined at the baseline and every 4 weeks. Total body water (TBW) was determined by the tritiated water technique at the baseline and 12 weeks after dieting. Extracellular water (ECW) was determined by 77Br space. Routine serum chemistries were obtained at 2-week intervals. Analysis of variance indicated no significant differences in the PRA and SA between the two diets. At the baseline, while on a self-selected 150 mEq sodium diet, there was a 3- to 4-fold increase in PRA after 2 h of ambulation (supine PRA 0.83 +/- 0.22 increased to 3.41 +/- 0.96 ng/ml/h). After the hypocaloric diets were instituted, the absolute values for PRA in the supine and upright positions declined. However, the magnitude of the postural response (3- to 4-fold increase) remained unchanged during the 12 weeks of weight reduction. There was no decline in the absolute values for supine or upright SA, during the entire study. Weight loss was significant (from 102.56 +/- 6.0 to 81.7 +/- 3.7 kg; P less than .001) and was accompanied by a mean +/- SE reduction in the TBW of 3.01 +/- 0.88 liters (P less than .011).(ABSTRACT TRUNCATED AT 250 WORDS)

Improved models for determination of body fat by in vivo neutron activation.

In the present study, two different models of body composition, based on data obtained by nuclear techniques are used. Total body nitrogen, calcium, and chlorine were obtained by total body neutron activation. Total body chlorine was used to estimate extracellular water, and total body calcium to determine bone mineral and extracellular solids. Total body potassium was measured by whole body counting to obtain the body cell mass. In addition, total body water was measured by the tritium dilution technique. It was found that either model can be used equally well to measure total body fat in normal subjects. Estimation of body fat as the difference between body weight and the sum of total body nitrogen (protein), total body water, and bone ash (model 1) appears to have an advantage over model 2, which uses body cell mass, extracellular water, and extracellular solids, particularly for patients with metabolic disorders. This advantage is partly due to the fact that the parameter protein (total body nitrogen) is less affected in metabolic disorders than the more labile total body potassium. The closely correlated results obtained with the two models based on nuclear measurements support the conclusion that these techniques provide reliable measurements of total body fat.

An improved calibration for the in vivo determination of body nitrogen, hydrogen, and fat.

Additional investigation of the authors' original technique for measuring total body nitrogen by prompt gamma neutron activation has demonstrated the need for certain changes in the calibration procedures in order to apply the method to studies of patients with abnormal metabolism. In the present technique, total body nitrogen, hydrogen, and fat were derived, simultaneously, from data obtained by neutron capture gamma-ray analysis combined with the measurements of body weight, total body water, and total body calcium. In this improved calibration technique total body nitrogen is more accurately measured, not only in normal subjects, but also in obese subjects and in patients with marked changes in hydration, such as cancer patients. The fat values calculated do not rely on a fixed relationship of total body water or total body potassium with lean body mass as in the previous studies, but are calculated as the difference between body weight and the sum of body water, protein and bone mineral ash. This improved technique has been applied to the study of three groups of subjects, the general population with a normal weight distribution and two extremes represented by obese and cancer patients.

Effects of hypophysectomy on tissue alkaline phosphatase in the rat.

To examine the effect of hypophysectomy on tissue alkaline phosphatase (EC 3.1.3.1) activity, studies were performed on mature rats. Hypophysectomy reduced the alkaline phosphatase activity in duodenum, kidney, and femoral bone. The decrease of alkaline phosphatase was greater in the intestine than in other organs studied. Intestinal alkaline phosphatase fell rapidly within 4 days post-hypophysectomy. In contrast, bone and kidney alkaline phosphatase activity had an initial lag period for 4 days after the hypophysectomy. Serum alkaline phosphatase also declined after hypophysectomy. The bone isoenzyme of serum alkaline phosphatase was reduced to a greater extent than the intestinal isoenzyme.

Determinants of bone mass in postmenopausal women.

Eighty white women, mean age 52 years, within one to six years postmenopausal, were studied to examine the relationship of various factors to bone mass. Forty-four of the women had annual measurements of bone mass, so that the rate of bone loss could be determined. Bone mass was measured by total body neutron activation analysis and photon absorptiometry of the distal radius (total body calcium [TBCa] and bone mineral content [BMC], respectively). Breast-feeding and pregnancy were noted to be associated with higher bone mass; those with lower BMC and/or TBCa tended to have higher serum alkaline phosphatase levels, lower testosterone levels, and more years since the cessation of menses. The rate of bone loss from the radius was greater in those with higher parathyroid hormone levels; those with reduced dietary intake of calcium and lower 25-hydroxyvitamin D levels had a greater rate of loss of TBCa.

Evaluation of body composition and nitrogen content of renal patients on chronic dialysis as determined by total body neutron activation.

Total body protein (nitrogen), body cell mass (potassium), fat, and water were measured in 15 renal patients on maintenance hemodialysis (MHD). Total body nitrogen was measured by means of prompt gamma neutron activation analysis; total body water was determined with tritium labeled water; total body potassium was measured by whole body counting. The extracellular water was determined by a technique utilizing the measurement of total body chloride and plasma chloride. When compared with corresponding values of a control group of the same age, sex, and height, the protein content, body cell mass, and total body fat of the MHD patients were within the normal range. The only significant change was an increase in the extracellular water/body cell mass ratio in the male MHD patients compared to the controls. The lack of significant difference of the nitrogen values of the MHD patients compared to matched controls suggests that dialysis minimizes any residual effects of uremic toxicity or protein-calorie malnutrition. These findings further suggest that there is a need to reevaluate the traditional anthropometric and biochemical standards of nutritional status for MHD patients. It was concluded that it is particularly important to measure protein stores of MHD patients with low protein intake to ascertain nutritional status. Finally, in vivo measurement of total body nitrogen and potassium for determination of body composition provides a simple, direct, and accurate assessment of the nutritional status of MHD patients.