A specific radioimmunoassay for osteocalcin with advantageous species crossreactivity

Patterson-Allen, Patricia; Brautigam, Cathy E.; Grindeland, R. E.; Asling, C. Willet; Callahan, Paul X.

doi:10.1016/0003-2697(82)90309-8

Cited by 56 publications

(17 citation statements)

References 8 publications

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“…procedures To validate the rat osteocalcin assay (Patterson-Allen et al, 1982) for use with bear samples, osteocalcin was purified from black bear cortical bone by modification of a previously described method (Colombo et al, 1993). The bear bone was broken into small fragments, defatted with a mixture of three parts hexane and two parts isopropanol, and lyophilized.…”

Section: Black Bear Osteocalcin Purification and Radioimmunoassaymentioning

confidence: 99%

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Parathyroid hormone may maintain bone formation in hibernating black bears (Ursus americanus) to prevent disuse osteoporosis

Donahue

Galley

Vaughan

et al. 2006

Journal of Experimental Biology

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SUMMARY Mechanical unloading of bone causes an imbalance in bone formation and resorption leading to bone loss and increased fracture risk. Black bears(Ursus americanus) are inactive for up to six months during hibernation, yet bone mineral content and strength do not decrease with disuse or aging. To test whether hibernating bears have biological mechanisms to prevent disuse osteoporosis, we measured the serum concentrations of hormones and growth factors involved in bone metabolism and correlated them with the serum concentration of a bone formation marker (osteocalcin). Serum was obtained from black bears over a 7-month duration that included periods of activity and inactivity. Both resorption and formation markers increased during hibernation, suggesting high bone turnover occurred during inactivity. However, bone formation appeared to be balanced with bone resorption. The serum concentration of parathyroid hormone (PTH) was higher in the hibernation(P=0.35) and post-hibernation (P=0.006) seasons relative to pre-hibernation levels. Serum leptin was lower (P<0.004)post-hibernation relative to pre-hibernation and hibernation periods. Insulin-like growth factor I (IGF-I) decreased (P<0.0001) during hibernation relative to pre-hibernation and reached its highest value during remobilization. There was no difference (P=0.64) in 25-OH vitamin D between the three seasons. Serum osteocalcin (bone formation marker) was significantly correlated with PTH, but not with leptin, IGF-I or 25-OH vitamin D. Osteocalcin and PTH were positively correlated when samples from all seasons were pooled and when only hibernation samples were considered, raising the possibility that the anabolic actions of PTH help maintain bone formation to prevent disuse osteoporosis. Prostaglandin E2 (PGE2)release from MC3T3 osteoblastic cells was significantly affected by treatment with bear serum from different seasons (i.e. hibernation versus active periods). The seasonal changes in PGE2 release showed trends similar to the seasonal changes in serum IGF-I. Since both PGE2 and IGF-I are associated with collagenous bone formation, it is possible that seasonal changes in a circulating factor influence IGF-I levels in vivo in bears and PGE2 release in osteoblastic cells in vitro. The significant decrease in serum leptin following arousal from hibernation may promote bone formation during remobilization, assuming there is a similar decrease in intracerebroventricular leptin. These findings support the idea that seasonal changes in the concentration of circulating molecules help regulate bone formation activity and may be important for preventing disuse osteoporosis in bears.

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Section: Black Bear Osteocalcin Purification and Radioimmunoassaymentioning

confidence: 99%

“…Highly purified bear osteocalcin and bear serum were assayed by radioimmunoassay (Patterson-Allen et al, 1982). The antibody was guinea-pig anti-rat osteocalcin and tracer was 125 I-labeled rat osteocalcin.…”

Section: Black Bear Osteocalcin Purification and Radioimmunoassaymentioning

confidence: 99%

Parathyroid hormone may maintain bone formation in hibernating black bears (Ursus americanus) to prevent disuse osteoporosis

Donahue

Galley

Vaughan

et al. 2006

Journal of Experimental Biology

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“…The serum was stored by freezing at -80°C, and the concentrations of bone metabolism markers {OC, type I procollagen carboxyterminal propeptide (PICP) as bone formation markers and ICTP as a bone resorption marker} were determined. The measurements were performed by RIA (doubleantibody method) using commercial kits, OC (DiaSorin Inc., Stillwater, MN) [29] and PICP/ICTP (Orion Diagnostica, Oulunsalo, Finland) [32]. Lepage et al (1990) reported on the validity of the commercial kit for horse OC [20].…”

Section: Measurement Of the Bone Metabolism Markersmentioning

confidence: 99%

Clinical Usefulness of the Measurement of Bone Mineral Content by Radiographic Absorptiometry in the Young Thoroughbred

Kobayashi

Ando

Kaneko

et al. 2007

JES

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The purpose of this investigation was to evaluate the clinical usefulness of the total radiographic bone aluminum equivalency (RBAE) Racehorses undergo very strenuous training during the growth period from the age of about 20 months for the racing season that begins in the summer of their third year after birth. In the growth period, bone metabolism is affected by both the growth and training; both bone resorption and bone formation are enhanced, and bones grow with increases in bone mineral content (BMC) due to accelerated bone remodeling [1,10,12,37]. The measurement of changes in BMC in training is important for the evaluation of the training effect on the skeleton and growth. BMC is generally measured by mineralometry, and various methods including the radiographic bone aluminum equivalency (RBAE) method [22], ultrasound method [18], computed tomography (CT) method [35], dual-energy x-ray absorptiometry (DEXA) method [8], and dual photon absorptiometry (DPA) method [23] have been devised and employed on horses. The RBAE method is a type of Radiographic Absorptiometry (RA). It has been reported to take a short time for measurement, requires no general anesthesia or sedation, costs less than other methods and is non-invasive [22]. This method is not easily applied to the measurement of quantitative changes. Therefore, the total RBAE method as a modified method in consideration of volume changes has been proposed [26]. It has been reported that the total RBAE method is a reliable tool for measurement of BMC in horses [19].The BMC immediately below the central nutritional foramen of the third metacarpal bone (McIII) using the total RBAE method has been reported to be related to bone disorders of the limbs [27].

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“…Previous work on cross-reactivity of polyclonal antibodies against bovine osteocalcin has shown that they cross-react, not only with human, but also with osteocalcin from monkey, horse, goat, cat and elk (20,21).…”

Section: Biochemistry and Molecular Biology Internationalmentioning

confidence: 99%

Production and characterisation of monoclonal and polyclonal amtobpdoes agaomst bovine osteocalcin

et al. 1997

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SUMMARY: Monoclonal and polycl0nal antibodies were produced against osteocalcin purified from bovine bone. The antibodies were used for the development of a two-site ELISA. A monoclonal antibody recognising residues 15-49 of bovine osteocatcin generated by cleavage with 70% formic acid was used as capture antibody and a biotinylated polyclonal antibody recognising the same peptide was used for detection. The two-site assay did not detect any tryptic peptides of bovine osteocalcin (residues 1-19, 20-43, and 45-49) nor did it detect the peptide 1-14 or the synthetic peptide 37-49. Cross-reaction was observed for osteocalcin in plasma from sheep, goat, chicken and human but not for osteocalcin from rat and pig.

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A specific radioimmunoassay for osteocalcin with advantageous species crossreactivity

Cited by 56 publications

References 8 publications

Parathyroid hormone may maintain bone formation in hibernating black bears (Ursus americanus) to prevent disuse osteoporosis

Parathyroid hormone may maintain bone formation in hibernating black bears (Ursus americanus) to prevent disuse osteoporosis

Clinical Usefulness of the Measurement of Bone Mineral Content by Radiographic Absorptiometry in the Young Thoroughbred

Production and characterisation of monoclonal and polyclonal amtobpdoes agaomst bovine osteocalcin

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