John K. Scariano

Serum levels of cross-linked N-telopeptides and aminoterminal propeptides of type I collagen indicate low bone mineral density in elderly women

Levels of aminoterminal propeptide (PINP) and the cross-linked carboxyterminal (ICTP) and aminoterminal (NTx) telopeptides of type I collagen were determined in the sera of 202 healthy women, 60-90 years of age, and correlated with densitometric measurements of their lumbar spine, femoral neck, and total body regions. Total skeleton bone mineral density (BMD) was associated most strongly with serum NTx levels (r = -0.51, p < 0.0001). Serum NTx and PINP levels correlated with BMD at all regions measured and were significantly increased in osteopenic women (16.0 vs. 13.3 nmol bone collagen equivalents/L for NTx, p = 0.0006 and 46.5 vs. 40.9 micrograms/L for PINP, p = 0.02). Elevations of NTx and PINP over a 3 year interval correlated with decreases in BMD measured at the femoral neck and to increases in serum concentration of alkaline phosphatase activity and ICTP. Serum levels of ICTP, NTx, PINP, ICTP, estrone, and alkaline phosphatase activity were correlated and serum NTx values were related to circulating thyroxine and intact parathyroid hormone levels. NTx and PINP levels were significantly decreased in the sera of women receiving estrogen replacement therapy (12.0 vs. 14.8 nmol bone collagen equivalents/L for NTx, and 35.5 vs. 45.2 micrograms/L for PINP, p < 0.001). Multiple regression analysis indicated that 42% of the variability observed in total skeletal BMD could be explained by age, weight, and serum levels of NTx and estrone. Within this model of covariance, the serum NTx level alone accounted for 10.1% of total body BMD variability.

Rickets in Nigerian children: response to calcium supplementation

In a previous study of rachitic children in Jos, Nigeria we concluded that inadequate dietary intake of calcium was the primary contributing factor to the development of their rickets. The objective of the present study was to determine the effect of calcium supplementation in 10 children with radiographically and biochemically proven rickets from the same geographical area. Rachitic children were provided with calcium supplements of 1000 mg/day for a period of 3 months. Serum and urine samples were obtained at baseline and at 24 hours, 1 week, 4 weeks, and 12 weeks after initiation of supplementation. Serum calcium, phosphorus, alkaline phosphatase, intact parathyroid hormone, 25-hydroxyvitamin D, and 1,25-dihydroxyvitamin D were measured at each time point. Dietary recalls obtained at two separate times were used to estimate usual daily intakes of calcium and phosphorus. Ten non-rachitic age-matched controls from the same geographical area were recruited for comparison. Nine of 10 rachitic subjects had radiographic evidence of healing after 3 months of calcium therapy. Although serum calcium concentrations returned to control levels, other biochemical data indicated that the rickets of these subjects may have been multifactorial in aetiology, pointing to a possible defect in the synthesis of 25-hydroxyvitamin D.

Estrogen receptors alpha (ESR1) and beta (ESR2) are expressed in circulating human lymphocytes.

Bone marrow thymocytes in part mediate the bone-preserving effects of estrogen by decreasing their production of osteoclast growth factors such as interleukin-1 and -6 and tumor necrosis factor alpha in the presence of physiological amounts of estradiol. Although several in vitro studies implicate the T-lymphocyte as a candidate mediator of estrogen signaling in the skeleton, whether these cells or any lymphocytes ordinarily express one or both nuclear estrogen receptors was previously unresolved. The purpose of our investigation was therefore to ascertain, by using real-time PCR, immmunoblotting, and cytometric techniques, if any of the nuclear estrogen receptors could be detected in normal peripheral blood mononuclear cells (PBMNC) collected from healthy volunteers. The results of immunoblotting experiments revealed that both estrogen receptor alpha (ESR1) and beta (ESR2) proteins are expressed in nuclei, but not in the cytoplasm of PBMNC harvested from all of the 15 healthy male and female volunteers (aged 23–50 years) we tested. PBMNCs contained mRNA coding for the two major full-length isoforms of ESR2 and the expression of ESR2 protein was localized within a lymphocyte subpopulation by cytometric analysis. Our data provide further evidence that lymphocytes and monocytes are responsive to estrogen and underscore its importance in modulating the immune response, as well as the vascular and skeletal health of men and women.

Serum leptin levels, bone mineral density and osteoblast alkaline phosphatase activity in elderly men and women

Although primarily secreted by adipose cells, leptin, a polypeptide hormone that influences body weight, satiety and lipid metabolism, and its receptor are also expressed in human osteoblasts. Leptin plays a role in the central, hypothalamic modulation of bone formation, as well as locally within the skeleton by enhancing differentiation of bone marrow stroma into osteoblasts and inhibiting its differentiation into osteoclasts and adipocytes. The purpose of this investigation was to compare serum leptin values in 100 postmenopausal women (age 62-97) and 31 men (age 72-92) to bone mineral density (BMD) measurements made by dual X-ray absorptiometry and additionally to biochemical markers of bone resorption and formation, including crosslinked collagen N-telopeptides (NTx), aminoterminal extension procollagen propeptides (PINP) and bone-specific alkaline phosphatase (bAP). The circulating level of leptin directly correlated with body mass index (BMI) (r=0.61-0.78, P<0.001) and was modestly, but significantly and positively associated with bAP activity (r=0.24-0.33, P<0.01) in the sera of men and women after adjustment for BMD, age and BMI. The association of circulating leptin levels with bAP, a specific marker of osteoblast activity suggests that leptin levels influence osteoblast activity in vivo in elderly women and men.

Critical differences in the serial measurement of three biochemical markers of bone turnover in the sera of pre- and postmenopausal women.

OBJECTIVES The purpose of this investigation was to quantify the biologic, day-to-day variability and critical differences in serum levels of crosslinked collagen N-telopeptides (NTx), procollagen aminoterminal extension propeptides (PINP) and bone specific alkaline phosphatase (bAP) in healthy women. DESIGN AND METHODS Seven blood samples were collected from 12 pre- and 15 postmenopausal women over 4 to 6 months. NTx, PINP and bAP levels were determined utilizing enzyme- and radioimmunoassay techniques. RESULTS The within-subject coefficient of variation (C.V.) in serum bAP, NTx and PINP levels was 7.1, 10.6 and 12.4% respectively. These variances did not differ significantly among premenopausal women when compared with postmenopausal subjects. Combining terms for analytical and biologic variability revealed that a critical difference between 2 successive serial measurements is 24% for bAP, 34% for NTx and 38% for PINP. CONCLUSION Circulating levels of NTx, PINP and bAP are stable over time periods of several months, allowing for the determination of significant changes in skeletal metabolism of women.

Serum levels of the pyridinoline crosslinked carboxyterminal telopeptide of type I collagen (ICTP) and osteocalcin in rachitic children in Nigeria

OBJECTIVES We measured the levels of the pyridinoline crosslinked carboxyterminal telopeptide of type I collagen (ICTP) and osteocalcin (OC) in the serum of 12 rachitic and 27 healthy Nigerian children, and compared the performance of these relatively new markers of bone metabolism with established laboratory parameters of skeletal disease. DESIGN AND METHODS Active rickets was diagnosed on the basis of clinical and biochemical criteria. Serum calcium and phosphorus concentration and alkaline phosphatase activity were determined using clinically accepted methods. Radioimmunoassay was performed to quantify parathyroid hormone, 1-,25-dihydroxyvitamin D, OC, and ICTP. RESULTS The rachitic children had statistically significant serum elevations of ICTP and osteocalcin as compared with age- and sex-matched controls. Serum levels of ICTP correlated with alkaline phosphate activity. CONCLUSIONS As a marker of abnormal bone metabolism, ICTP performs at least as well as alkaline phosphate. ICTP and OC are valuable additions to the growing repertoire of bone markers.

Bone mineral density and serum levels of aminoterminal propeptides and cross-linked n-telopeptides of type I collagen in elderly men

Serum levels of aminoterminal extension propeptides (PINP), the carboxyterminal telopeptide (ICTP), and the cross-linked N-telopeptides (NTx) of type I collagen were determined in 78 healthy, elderly men aged 76 +/- 5 years in 1993 and 1996 and compared with bone mineral density (BMD) measurements of their lumbar spine, femoral neck, and total body regions made using dual X-ray absorptiometry. Compared with 11 men who had normal lumbar spine (SBMD) and femoral neck BMD (NBMD), 13 of the subjects with SBMD and NBMD classified as osteopenic by t-score criterion had higher mean serum levels of PINP and alkaline phosphatase activity, but these increases were not statistically significant at the 95% confidence interval. In osteopenic men, a correlation between SBMD and NTx was detected (r = -0.66, p = 0.01). Within the entire population, the serum NTx level correlated with NBMD (r = -0.26, p < 0.05) and PINP (r = +0.63, p < 0.0001), and the change in the circulating concentration of PINP over the 3 year interval correlated with the magnitude of change in total body BMD (r = -0.28, p = 0.02), NBMD (r = -0.24, p = 0.05), and SBMD (r = -0.36, p = 0. 03) as well as with the change in serum NTx levels (r = 0.43, p < 0. 001). The change in the circulating ICTP level was also related to the change in NBMD (r = -0.24, p = 0.01). Together, weight and the serum PINP level accounted for 25% of total body BMD variance in elderly men. These results indicate that larger populations of men and women should be screened over longer time intervals to explore the value of serial measurement of serum collagen metabolites in predicting bone loss in the spine and hip.

Rickets and Protein Malnutrition in Northern Nigeria

The aim of the study was to explore the relationship between protein nutritional status and the development of rickets in children living in northern Nigeria. The diagnosis of rickets in 16 children between the ages of 10 months and 7 years was confirmed using established, and recently developed clinical and biochemical parameters. Twenty-seven children devoid of skeletal stigmata were age- and sex-matched to the rachitic patients. A battery of clinical laboratory and anthropometric measurements designed to assess calcium homeostasis, skeletal growth, the extent of bone remodeling or resorption, and protein nutritional status were performed on all subjects. Our central finding was that although the rachitic children were moderately malnourished, their protein nutritional status was significantly better as measured by the serum prealbumin concentration (15.4 v. 12.5 mg/dl, P = 0.0012) when compared with the severely malnourished children who were devoid of any indication of rickets. This may be due, in part, to the fact that actively growing children are more likely to develop rickets than are children whose linear growth is impeded. Unexpectedly, we found that the mean concentrations of serum 1,25-dihydroxyvitamin D in both the rachitic and control group were higher than any values for the active vitamin D metabolite previously reported in the literature.

Diagnostic efficacy of serum cross-linked N-telopeptide (NTx) and aminoterminal procollagen extension propeptide (PINP) measurements for identifying elderly women with decreased bone mineral density.

The purpose of this study was to determine the diagnostic sensitivity, specificity, predictive value and overall efficiency of serum cross-linked N-telopeptides of bone collagen (NTx) and aminoterminal procollagen extension propeptide (PINP) measurements for identifying women with decreased spine, femoral neck and total body bone mineral density (BMD). Serum NTx and PINP levels and dual X-ray absorptiometry were performed on 196 healthy elderly women, aged 60-90 years. Twelve women were classified as having decreased BMD on the basis of regional and total skeletal densitometric values that were 1.5 to 2.5 standard deviations (SD) below the respective, age-stratified means and were compared with184 women with BMD values greater than 1.5 SD below the mean. The results of receiver operating characteristic analysis revealed that a cutoff level of more than 15.0 nmol BCE/L for serum NTx, as measured by the Osteomark assay (Ostex International, Seattle WA USA) was associated with a 100% sensitivity and 70% specificity rate for identifying postmenopausal women with low BMD. The positive likelihood ratio was 3.3 and the negative predictive value was 1.0 using the 15.0 nmol decision level for NTx. The overall diagnostic efficiency of a single NTx measurement for identifying women with low BMD was 89%. A cutoff level of > 45.0 mug/L for PINP as measured by the Orion Diagnostica RIA assay (Espoo, Finland) had a diagnostic sensitivity of 83% and specificity of 64% for identifying women with decreased BMD. The positive likelihood ratio was 2.3, the negative predictive value 0.98 and the overall diagnostic efficiency 73% using the 45.0 mug/L decision level for PINP. These results warrant future studies using larger populations that are inclusive of more women with low bone mineral density.

Calcium supplements increase the serum levels of crosslinked N-telopeptides of bone collagen and parathyroid hormone in rachitic Nigerian children☆

OBJECTIVES Biochemical markers of bone turnover were measured in the sera of 16 controls and 10 children with calcium-deficiency rickets, during a 12-week course of calcium supplementation (1 g CaCO3/d) that was effective in healing the bone lesions of the rachitic children. DESIGN AND METHODS Serum levels of crosslinked N-telopeptides of bone collagen (NTx), parathyroid hormone (PTH), alkaline phosphatase (ALP), and urinary deoxypyridinoline (LP) were assayed at baseline and during the course of calcium therapy and compared with data of the 16 non-rachitic controls. RESULTS Calcium therapy suppressed serum NTx and PTH levels in the rachitic children within 24 h; however, after the first week, PTH and NTx levels increased to the extent that at 12 weeks both were elevated when compared with controls or to baseline levels. Serum levels of NTx and PTH were correlated in the controls and experimental subjects (r = 0.63, p < 0.001). CONCLUSIONS The rate of bone resorption, as estimated by serum NTx concentration, is increased during the healing of rachitic lesions.