bluuebunny/crossref_metadata_2025_split huggingface dataset:
1,25-Dihydroxyvitamin D reduces parathyroid hormone receptor number in ROS 17/2.8 cells and prevents the glucocorticoid-induced increase in these receptors: Relationship to adenylate cyclase activation
Authors: Dr. Titus Louisa , Jackson Elizabeth , Nanes Mark S. , Rubin Janet E. , Catherwood Bayard D. 
Published: February 2010

Abstract
We have previously shown that 1,25-dihydroxyvitamin D [1,25-(OH)2D3] and glucocorticoid modulate adenylate cyclase activation by PTH in osteoblast-like cells. Here we examine whether steroid effects on PTH receptor density explain the modulation of PTH action. Receptor assays were performed on late logarithmicphase monolayers of ROS 17/2.8 cells using human PTH-like peptide (hPLP) as radioligand. Kd and receptor density were computed from competition of tracer amounts of [125I-Tyr36]hPLP-(1–36) with unlabeled hPLP-(1–36) (0.1–30 nM). Steroid treatment had little or no effect on affinity for ligand. Pretreating cells with 10 nM 1,25-(OH)2D3 for 48 h decreased PTH receptor number to 17% of control values. Treating cells with 10 nM of the glucocorticoid triamcinolone acetonide (TRM) increased receptor number 10-fold, but simultaneous treatment with 1,25-(OH)2D3 (10 nM) completely prevented this receptor increase. Steroid effects required 13–18 h of treatment. Dose-response relationships for steroid modulation, determined from binding at 0.17 nM radioligand, indicated an EC50 of 0.3 nM for glucocorticoid augmentation of PTH receptor number and 0.02 nM for 1,25-(OH)2D3 reduction of receptor number in the presence or absence of the maximum TRM effect. The initial rate of cAMP production by receptor-saturating concentrations of PTH was 11,500 molecules per receptor per minute in untreated cells, comparable to reported turnover numbers for mammalian adenylate cyclase. Control experiments were validated measuring cAMP in intact cells as an indicator of adenylate cyclase activity. Cyclic AMP production was reduced 63% by 1,25-(OH)2D3 (10 nM) treatment. Glucocorticoid (10 nM) enhanced cAMP production twofold but reduced cAMP generation per receptor by 80%. We conclude that 1,25-(OH)2D3 reduction of PTH receptor number modulates ROS cell responses to PTH and prevents glucocorticoid-enhanced PTH action. The data also suggest that other components of the adenylate cyclase complex limit amplification of the PTH signal when PTH receptor number is elevated by glucocorticoid treatment.
Inhibition of 1,25-(OH)2D3- and 24,25-(OH)2D3-dependent stimulation of alkaline phosphatase activity by A23187 suggests a role for calcium in the mechanism of vitamin D regulation of chondrocyte cultures
Authors: Schwartz Z. , Langston G.G. , Swain L.D. , Boyan Barbara D. 
Published: February 2010

Abstract
This study used the ionophore, A23187, to examine the hypothesis that the regulation of alkaline phosphatase and phospholipase A2 activity by vitamin D3 metabolites in cartilage cells is mediated by changes in calcium influx. Confluent, fourth-passage cultures of growth zone and resting zone chondrocytes from the costochondral cartilage of 125 g rats were incubated with 0.01-10 μM A23187. Specific activities of alkaline phosphatase and phospholipase A2 were measured in the cell layer and in isolated plasma membranes and matrix vesicles. There was an inhibition of alkaline phosphatase specific activity at 0.1 μM A23187 in resting zone cells and at 0.1 and 1 μM in growth zone chondrocytes. At these concentrations of ionophore, the 45Ca content of the chondrocytes was shown to increase. Both the plasma membrane and matrix vesicle enzyme activities were inhibited. There was no effect of ionophore on matrix vesicle or plasma membrane phospholipase A2 in either cell type. In contrast, alkaline phosphatase activity is stimulated when growth zone chondrocytes are incubated with 1,25-(OH)2D3 and in resting zone cells incubated with 24,25-(OH)2D3. Phospholipase A2 activity is differentially affected depending on the metabolite used and the cell examined. Addition of ionophore to cultures preincubated with 1,25-(OH)2D3 or 24,25-(OH)2D3 blocked the stimulation of alkaline phosphatase by the vitamin D3 metabolites in a dose-dependent manner. The effects of ionophore were not due to a direct effect on the membrane enzymes since enzyme activity in isolated membranes incubated with A23187 in vitro was unaffected. These results suggest a role for calcium in the action of vitamin D metabolites on chondrocyte membrane enzyme activity but indicate that mechanisms other than merely Ca2+ influx per se are involved.
Depression of osteoblastic activity in immobilized limbs of suckling rats
Authors: Weinreb Miron , Rodan Gideon A. , Thompson David D. 
Published: February 2010

Abstract
Recently we characterized the immobilization-related osteopenia in adult rats and showed that it is caused by increased bone resorption and decreased bone formation (Weinreb et al. 1989 Bone 10:187). To assess the effect of age on disuse osteopenia, this study investigated the effects of immobilization on bone turnover in very young, suckling rats. The 15-day-old rats underwent unilateral hind limb immobilization by sciatic neurectomy; the contralateral limb was left intact and served as control. Experimental or sham-operated animals were killed after 0, 2, 4, or 12 days postsurgery. Dry, fat-free weight and ash weight were determined in both femora, and both tibiae were subjected to static and dynamic histomorphometry. Immobilization caused a progressive deficit in bone mass in the immobilized limb compared to the contralateral intact limb but did not affect femoral longitudinal growth. The total mineral content in the immobilized femora was 13.6% less than that in the intact limb by day 12. Concomitantly, tibial cancellous bone area and perimeter declined in the immobilized limb by 37.3 and 32.2%, respectively. This reduction in bone mass in the tibiae of immobilized limbs was associated with increased bone resorption, expressed as osteoclast perimeter, number of osteoclasts per mm surface, and number of osteoclasts per mm2 tissue area. Bone formation was reduced as a result of impaired osteoblast activity as evidenced by (1) decreased endocortical and trabecular mineral apposition rate; (2) reduced trabecular mineral formation rate; (3) decreased percentage of ash of the femoral dry weight; and (4) increased volume of unmineralized osteoid in the tibial metaphysis. These data indicate that osteoblasts in very young rats are sensitive to mechanical disuse and exhibit impaired function within a short period after the onset of immobilization. They also suggest that mechanical unloading inhibits mineralization to a greater extent than matrix deposition, leading to the production of hypomineralized bone.
Effects of systemic aluminum on the resolution of a uremic and dietary phosphorus-dependent model of uremic osteomalacia in rats
Authors: Lieuallen Warren G. , Dr. Weisbrode Steven E. 
Published: February 2010

Abstract
We have developed a model of osteomalacia that is dependent on both uremia and the feeding of a diet low in phosphorus and that can be reversed by subsequent dietary phosphorus repletion. The objectives for this study were to use this model to investigate the role of aluminum (Al) in both the induction and resolution of osteomalacia. Adult male Sprague-Dawley rats were five-sixths nephrectomized and fed either low or normal dietary phosphorus, both with and without intraperitoneal Al injections. Uremic rats fed low phosphorus developed osteomalacia characterized by increased osteoid surface, volume, and thickness and osteoid maturation time and decreased mineralizing surface. Al-treated uremic rats fed low phosphorus were similarly affected, developing increased osteoid volume and thickness and osteoid maturation time and decreased osteoblastic surface, mineralizing surface, and bone formation rate. In addition, they had a significantly increased Al-positive surface. Al-treated uremic rats fed normal phosphorus had only increased osteoid thickness and aluminum-positive surface and decreased osteoblastic surface. Osteomalacic rats continuously treated with Al during the induction and phosphorus repletion stages had increased growth plate thickness, osteoid volume and thickness, and Al-positive surface and decreased osteoblastic and mineralizing surface. Mineralization in these rats was impaired to such a degree that no detectable double labels were present. Osteomalacic rats treated with Al during the induction phase but not during phosphorus repletion had increased osteoid surface and volume and Al-positive surface and decreased osteoblastic and mineralizing surface. Double labels were not detectable in these rats, either. These results indicate that the osteomalacia induced in uremic rats fed low phosphorus was similar in rats with and without Al treatment. However, Al prevented the reversal of this osteomalacia by phosphorus repletion, even when Al administration was discontinued. The impairment of mineralization by Al in this model of osteomalacia suggests that Al-positive bone surfaces are not an epiphenomenon in osteomalacia associated with uremia.
Solid-phase synthesis and biologic activity of human parathyroid hormone(1–84)
Authors: Goud Nagana A. , McKee Roberta L. , Dehaven Patrica A. , Huelar Elizabeth , Syed Mustaqim M. , Goud Rajeswari A. , Gibbons Susan W. , Fisher John E. , Levy Jay J. , Rodkey John A. , Bennett Carl , Ramjit Harri G. , Caporale Lynn H. , Caulfield Michael P. , Dr. Rosenblatt Michael 
Published: February 2010

Abstract
We have chemically synthesized the full-length, 84 amino acid, human parathyroid hormone (hPTH) on a &gt; 100 mg scale by the Merrifield solid-phase technique of stepwise peptide synthesis using a benzhydrylamine support. The peptide was purified by high-performance liquid chromatography and found to be &gt; 96% pure. The authenticity of the sequence of the synthetic peptide was confirmed by repetitive Edman degradation. Furthermore, tryptic digestion of hPTH generated the predicted fragments. The synthetic full-length hormone was evaluated for biologic activity in assays of PTH receptor binding and stimulation of adenylate cyclase activity (using bovine renal cortical membranes and rat and human bone cells). Synthetic hPTH(1–84) was found to be highly potent in binding to PTH receptors (Kb = 1–25 nM) and stimulating adenylate cyclase (Km = 1–14 nM). The availability of significant quantities of synthetic full-length hPTH and future analogs will permit widespread use in multiple in vitro and in vivo assays to delineate their spectrum of biologic properties. Available supplies of the synthetic hormone will also enable evaluation of the effectiveness of PTH antagonists at inhibiting the action of native sequence hormone at its receptors.
Evidence for interleukin-1β production by cultured normal human osteoblast-like cells
Authors: Keeting Philip E. , Rifas Leonard , Harris Steven A. , Colvard Douglas S. , Spelsberg Thomas C. , Peck William A. , Dr. Riggs Lawrence B. 
Published: February 2010

Abstract
To determine if bone cells produce interleukin-1β (IL-1β), a potent bone resorption-stimulating agent, we studied well-characterized, nearly homogeneous cultures of normal human osteoblast-like (hOB) cells. With four strains of such cells, vehicle-treated cultures produced minimal IL-1β (mean ± SEM, 1.3 ± 0.3 pg/ml per 106 cells per 24 h) and showed dose-dependent (r = 0.99) increases to 2.2 ± 0.7, 5.0 ± 0.9, or 17.8 ± 6.7 pg/ml, respectively, after treatment with lipopolysaccharide (LPS) at 3, 10, or 30 μg/ml (for increases after 10 and 30 μg/ml treatments, P &lt; 0.05). After treatment with tumor necrosis factor α (TNF-α at 10 U/ ml, IL-1β increased to 16.2 * 3.7 pg/ml (P &lt; 0.05). Neither 17β-estradiol nor bovine parathyroid hormone(1–34) (each at 10 nM), alone or in combination with LPS or TNF-α, affected IL-1β release. Northern blot analysis of total cellular RNA preparation revealed a single hybridization band at 1.9 kb when probed with a partially deleted cDNA for human IL-1β. The steady-state IL-1β mRNA levels showed a significant increase with LPS treatment and a lesser increase with TNF-α treatment in hOB cells. Moreover, TNF-α produced an even greater increase in IL-1 mRNA in HOBIT cells, a well-differentiated clonal cell line derived from normal hOB cells transfected with the SV40 large T antigen. We conclude that human cells of the osteoblast lineage produce IL-Iβ in response to well-recognized stimuli for IL-1 release from responsive tissue. Therefore, IL-1 may play an important role in the local regulation of bone remodeling.
Uptake of fluorine in cortical and trabecular bone
Authors: McNeill K.G. , Coote G.E. , Hitchman A.J.W. 
Published: February 2010

Abstract
Nuclear magnetic resonance techniques can measure the fluoride levels in bone of the finger after a patient has ingested F in the treatment of osteoporosis, but does uptake in cortical bone reflect uptake in the critical trabecular bone? Investigation has been made of the relative uptake of fluoride from drinking water into trabecular and cortical bone of the rat. For fine detail of the uptake of F into the femur and vertebra, microprobe techniques were used with a spatial resolution of 10 μm; for broader studies, treating the femur as representative of cortical bone and the vertebra as typical of trabecular bone, chemical techniques using spectroscopy and ion-selective electrodes were employed. The conclusion is that in the rat uptake of F by cortical bone is indicative of uptake by trabecular bone, and that therefore as a working hypothesis NMR measurement of F in the finger may be taken as reflecting uptake of F by trabecular bone.
A new approach to defining normal vertebral dimensions
Authors: Black Dennis M. , Cummings Steven R. , Stone Katie , Hudes Estie , Palermo Lisa , Steiger Peter 
Published: February 2010

Abstract
We developed a method for estimating the mean and standard deviation of ratios of normal vertebral heights from a sample that includes people with and without vertebral fractures. This method assumes that the measurements in normal vertebrae have a Gaussian distribution and that, for any vertebral level, the prevalence of abnormal measurements is less than 10%. Under these assumptions, normal values for nonfractured vertebrae can be estimated from several statistical properties of Gaussian distributions. We applied these methods to the lateral spinal radiographs of 2992 women aged 65–70 years who were recruited from population-based listings. The estimated means and standard deviations for ratios of dimensions in nonfractured vertebrae were very similar to those based on studies of premenopausal women. Our method may be useful for defining normal values from large populations that include normal and abnormal women, does not require x-rays of normal premenopausal women, avoids the potential biases of defining normality based on qualitative judgment, and can be applied to other types of physical and biochemical measurements.
Nonhypercalcemic 1,25-(OH)2D3 analogs potently induce the human osteocalcin gene promoter stably transfected into rat osteosarcoma cells (ROSCO-2)
Authors: Morrison Nigel A. , Eisman John A. 
Published: February 2010

Abstract
1,25-Dihydroxyvitamin D3[1,25-(OH)2D3] is the active hormonal form of vitamin D3 and has potent effects on bone and calcium regulation. Over the past decade it has become apparent that 1,25-(OH)2D3 has other effects on cellular proliferation that potentially could be developed for therapy in human malignancy. Since the hypercalcemic effects of 1,25-(OH)2D3 have limited that use in the human, novel nonhypercalcemic analogs of 1,25-(OH)2D3 have been synthesized. The molecular mechanism of this divergence in these antiproliferative and calcium-regulating actions is unexplained. We have previously examined the human bone-specific gene osteocalcin as a model of the molecular mechanisms of vitamin D action in bone and have shown that induction of the osteocalcin gene by 1,25-(OH)2D3 is mediated through an unique and complex palindromic region of the promoter similar to but distinct from those of other steroid hormone-responsive elements. Using an osteosarcoma cell line permanently transfected with the vitamin D-responsive promoter of the human osteocalcin gene linked to a “reporter” gene, we have shown that there is a dose-dependent induction of CAT activity by 1,25-(OH)2D3 and that the potencies of vitamin D metabolites and analogs are comparable to those found in other vitamin D bioassays. Furthermore, vitamin D analogs, including MC-903, 22-oxa-1,25-(OH)2D3, and Δ22–1,25S,26-trihydroxyvitamin D3, which effect cellular differentiation but lack hypercalcemic activity in vivo, exhibit osteocalcin promoter inductive actions virtually identical to those of 1,25-(OH)2D3. Consideration of these and other data support the hypothesis that the divergent effects of such analogs on differentiation and calcium homeostasis reflect pharmacokinetic differences in vivo rather than distinct 1,25-(OH)2D3-sensitive pathways.
Adenylate cyclase stimulating activity immunologically similar to parathyroid hormone-related peptide can be extracted from fetal rat long bones
Authors: Wolf Nicole Nijs-de , Pepersack Thierry , Corvilain Jacques , Karmali Rafik , Bergmann Pierre 
Published: February 2010

Abstract
We observed that culture medium conditioned with fetal rat long bones contained peptides immunologically related to the parathyroid hormone-related peptide of malignancy (PTHrP) and stimulated cyclic AMP production in canine renal cortical membranes. Because the adenylate cyclase stimulating activity (CSA) of the medium increased when bone resorption was stimulated, it was suspected that these peptides were stored in the matrix and released during the resorption process. In this work, we extracted the noncollagenous proteins of fetal rat long bones and found that the extract contained significant amounts of CSA. The biologic activity of the extract was abolished after trypsin digestion and eluted at 24 and 37 kD on filtration HPLC. The CSA of bone extract and of both HPLC peaks could be inhibited by 3–34 and 7–34 parathyroid hormone analogs. It was not blocked by an antiserum directed against the N-terminal region of parathyroid hormone, but it was significantly inhibited after an overnight preincubation with an antiserum directed against the 1–11 fragment of PTHrP. One band migrating at 18 kD could be visualized after SDS-PAGE electrophoresis of bone extract and immunoblotting with the anti-PTHrP antiserum. We conclude that an adenylate cyclase stimulator immunologically similar to PTHrP is present in the matrix of fetal rat long bones. Adenylate cyclase stimulating peptides of lower molecular weight found in bone-conditioned medium could be active fragments formed by proteolysis during the resorption process.
Iliac trabecular bone formation predicts radial trabecular bone density changes in type 1 osteoporosis
Authors: Hesp Richard , Arlot Monique E. , Edouard Claude , Bradbeer Jeremy N. , Meunier Pierre J. , Dr. Reeve Jonathan 
Published: February 2010

Abstract
In 28 patients with idiopathic or postmenopausal type 1 (spinal crush fracture) osteoporosis, resorption indices and dynamic measurements of trabecular bone formation based on in vivo tretracycline labeling in 7.5 mm transiliac biopsies have been compared with trends in radial cortical and trabecular bone density measured with computed tomography. Positive correlations were observed between trabecular bone density trends in the radius and indices of bone formation in the ilium. These were improved when one of the two resorption indices was included with a formation index in bivariate regressions. Marked interindividual variations in radial bone density trends were also seen in cortical bone. These correlated poorly with trends in trabecular bone. Weak negative relationships between cortical bone trends and indices relating to bone formation and resorption were observed, but a positive association was seen with single-labeled surfaces on iliac trabeculae. If, as has been suggested, there are periodic variations in bone formation, the results suggest that axial and peripheral trabecular bone density trends are synchronized in osteoporosis, perhaps in response to systemic factors, such as circulating hormones.
Interleukin-3-dependent hematopoietic stem cell lines capable of osteoclast formation in vitro
Authors: Hagenaars C.E. , Haas E.W.M. Kawilarang-de , van der Kraan A.A.M. , Spooncer E. , Dexter T.M. , Nijweide P.J. 
Published: February 2010

Abstract
Recently we reported that the osteoclast originates from the pluripotent hematopoietic stem cell. However, a detailed analysis of the progenitor and precursor stages of the osteoclast lineage is hard to perform with primary cultures of stem cells. In the present investigation interleukin-3 (IL-3) -dependent multipotent hematopoietic stem cells lines (FDCP-mix), which have many characteristics in common with freshly isolated hematopoietic stem cells, were assayed for their osteoclast formation capacity. FDCP-mix cell lines A4, C2GM, and 15S were cocultured with periosteum-free 17-day-old fetal metatarsal bones. The effects of culture time, medium composition, and addition of WEHI-3b-conditioned medium (an unpurified IL-3 preparation) on osteoclast formation were studied. 15S cells never differentiated into osteoclasts. Both A4 and C2GM cells were able to generate osteoclasts. Osteoclast formation was visualized by staining for tartrate-resistant acid phosphatase activity and confirmed by 45Ca release assays and electron microscopic studies. Medium supplemented with fetal calf serum clearly supported osteoclast formation from A4 cells better than medium supplemented with cock serum. The difference between fetal calf serum and horse serum is generally less pronounced. C2GM cells formed osteoclasts more readily and, generally, earlier than A4 under all culture conditions. WEHI-3b-conditioned medium addition increased the numbers of osteoclasts and their resorption activity. The coculture of stripped metatarsal bones with FDCP-mix cell lines therefore offers a model system with many possibilities for the study of osteoclastogenesis and its regulation.
Role of colony-stimulating factors in osteoclast development
Authors: Takahashi Naoyuki , Udagawa Nobuyuki , Akatsu Takuhiko , Tanaka Hirofumi , Shionome Manabu , Dr. Suda Tatsuo 
Published: February 2010

Abstract
Effects of various colony-stimulating factors (CSFs) [interleukin-3 (IL-3), granulocyte-macrophage CSF (GM-CSF), macrophage CSF (M-CSF), and granulocyte CSF (G-CSF)] on osteoclast-like cell formation were examined in two different culture systems: the one-step mouse marrow culture system and the two-step coculture system of mouse primary osteoblastic cells with the bone marrow cells collected from the colonies that formed in the methylcellulose in the presence of the CSFs. In the one-step mouse marrow cultures, none of the CSFs stimulated the formation of tartrate-resistant acid phosphatase (TRAP, a marker enzyme of osteoclasts)-positive multinucleated cells (MNCs). Furthermore, the CSFs other than G-CSF inhibited in a dose-dependent manner the TRAP-positive MNC formation induced by 1α-25-dihydroxyvitamin D3 [1α,25-(OH)2D3]. In contrast, when marrow cells were first cultured in semisolid methylcellulose in the presence of a CSF and the recovered marrow cells from the semisolid cultures were subsequently cocultured with primary osteoblastic cells in the presence of 1α,25-(OH)2D3, numerous TRAP-positive MNCs were formed. [125I]salmon calcitonin specifically bound to TRAP-positive cells formed in this two-step culture system. Over 90% of the TRAP-positive mononuclear cells and MNCs accumulated [125I]calcitonin. M-CSF was the most potent in inducing TRAP-positive MNCs, followed by GM-CSF, IL-3, and G-CSF in that order. No TRAP-positive cells were formed in the absence of either osteoblastic cells or 1α,25-(OH)2D3. These results clearly indicate that the CSFs, especially M-CSF, stimulate the growth of osteoclast progenitors, which differentiate into osteoclast-like cells in response to 1α,25-(OH)2D3 in the presence of osteoblastic cells.