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  • Articles  (293)
  • Articles: DFG German National Licenses  (293)
  • Nitrogen fixation  (148)
  • Bone  (145)
  • Springer  (293)
  • Cambridge University Press
  • 1980-1984  (191)
  • 1975-1979  (102)
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  • Articles  (293)
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  • Articles: DFG German National Licenses  (293)
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 33 (1981), S. 239-242 
    ISSN: 1432-0827
    Keywords: Bone ; Magnetic field ; IR light ; Light absorption constant ; Efficiency
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Enhancement of photovoltage has been found in bone and its two major constituents when placed in a magnetic field in standard Hall geometry. Infrared light has been used for carrier excitation. From data obtained in this manner, values for the light absorption constant, extinction constant, mass absorption constant, and efficiency have been estimated. All these parameters increase with the increase of magnetic field. A possible explanation for the observed phenomena is presented.
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  • 2
    ISSN: 1432-0827
    Keywords: Vitamin D ; Vitamin D deficiency ; Cartilage ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary To test the importance of 24-hydroxylation of vitamin D3 on bone mineralization, rat pups born to vitamin D-deficient females were given either 25-hydroxyvitamin D3 or 24,24-difluoro-25-hydroxyvitamin D3 for 16 days beginning at the time of weaning. Following such treatment analysis of blood samples revealed no detectable 24R,25-(OH)2D3 and 1,25-(OH)2D3 in the rats given the difluoro compound while revealing the expected 24,24-difluoro-25-hydroxyvitamin D3 and 24,24-difluoro-1,25-dihydroxyvitamin D3. The rats given 25-hydroxyvitamin D3 had the expected levels of 25-hydroxyvitamin D3, 24,25-dihydroxyvitamin D3, and 1,25-dihydroxyvitamin D3. Following sacrifice at day 17, postweaning bone mineralization and modeling were studied in long bones using histological methods. Bones taken from vitamin D-deficient rats at the beginning and end of the experimental period had lesions typical of rickets. These included wide growth plates, excessive amounts of osteoid, and metaphyseal fibrosis. Following treatment with either 25-hydroxyvitamin D3 or 24,24-difluoro-25-hydroxyvitamin D3, bone mineralization returned to normal. Growth plate widths and the amount of osteoid on bone surfaces were both substantially reduced and to a similar degree in both treatment groups. Normal cartilage core formation and trabecularization of the metaphyseal primary spongiosa were also restored to a similar degree in both groups. In effect, no difference was observed in any bone parameter studied between the 25-hydroxyvitamin D3- and the 24,24-difluoro-25-hydroxyvitamin D3-treated animals. These results provide strong evidence that 24-hydroxylation of the vitamin D molecule plays little or no role in the modeling and mineralization of bone.
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  • 3
    Electronic Resource
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    Calcified tissue international 33 (1981), S. 655-661 
    ISSN: 1432-0827
    Keywords: 1,25-Dihydroxyvitamin D3 ; 25-Hydroxyvitamin D3 ; 24,25-Dihydroxyvitamin D3 ; Receptors ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The hormonal metabolite of vitamin D3, 1,25-dihydroxyvitamin D3 [1,25(OH)2D3], exerts its biological effects by binding to a cytosolic receptor protein. Such a protein has been demonstrated in vitamin D3 target organs including fetal rat calvariae and more recently in rat osteogenic sarcoma cells. In this study we have compared the binding of 25-hydroxyvitamin D3 [25(OH)D3] and 24,25-dihydroxyvitamin D3 [24,25(OH)2D3] to that of 1,25-(OH)2D3 in fetal rat calvariae and osteogenic sarcoma (OS) cells. Sucrose density sedimentation, DNA-cellulose chromatography, and intracellular uptake studies have been employed to evaluate these interactions. In cytosol preparations from calvariae, [3H]-1,25(OH)2D3 bound to a 3.3S macromolecule and to a much greater extent to a 5.8S macromolecule while both [3H]25(OH)D3 and [3H]24,25(OH)2D3 bound to the 5.8S macromolecule. By incubating intact calvariae and OS cells with labeled metabolites and thus establishing binding intracellularly prior to cell disruption, we have found that the 3.3S protein which has high specificity for 1,25(OH)2D3 occurs inside the cells; the 5.8S protein, however, does not occur inside the cells but is generated after cell disruption. The [3H]-1,25(OH)2D3-receptor complex adsorbed to DNA-cellulose and was eluted from this affinity resin at 0.28M KCl. In contrast, [3H]25(OH)D3 and [3H]-24,25(OH)2D3 binding activity did not adsorb to DNA-cellulose. We conclude that, in contrast to the 3.3S protein, the 5.8S macromolecule does not fulfill receptor criteria but is rather generated by the experimental manipulation of the bone cells. Our data suggest that the vitamin D3 actions on bone are mediated only via the 3.3S receptor, and hence quantitative but not qualitative differences of the effects of the various metabolites are feasible.
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  • 4
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    Calcified tissue international 22 (1977), S. 303-313 
    ISSN: 1432-0827
    Keywords: Osteoclasts ; Sedimentation ; Isolation ; Cells ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract A method is presented for separating osteoclasts from the heterogeneous population of bone and marrow cells. Cell suspensions were prepared from femora of young rabbits by mechanical dispersion. The starting cell suspension typically contained only 1.0%±0.5 osteoclasts. Following an initial 45 min of unit gravity sedimentation in a lucite chamber osteoclasts were primarily distributed in fractions 2–5. A second 45-min sedimentation of these pooled fractions yielded cell suspensions containing greater than 30% osteoclasts (as much as a 50-fold increase over starting percentages). Linear scan analysis, however, revealed that osteoclasts accounted for 73.14%±0.58 of the cell colume. Subsequent in vitro experiments demonstrated linear incorporation of3H-leucine into TCA precipitable protein for cells comprising the osteoclast fraction. Concomitant radiautographs revealed radioactive label in isolated osteoclasts.
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  • 5
    ISSN: 1432-0827
    Keywords: Bone ; Diphosphonates ; Bone resorption ; Bone formation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Male rats weighing 100 g were assigned to groups and injected daily for 10 days with vehicle (control), 0.4, 2.0, 4.0, 10.0, or 20.0 mg/kg/day of ethane-1-hydroxy-1,1-diphosphonate (EHDP) or dichloromethylene diphosphonate (Cl2MDP). The proximal tibial metaphysis and epiphysis were assayed for changes in percentage of hard tissue and bone formation parameters. From the data, information about hard tissue resorption was deduced. All doses of Cl2MDP and doses of 2.0 mg EHDP/kg/day and greater caused significant increases in percentage of hard tissues with Cl2MDP being more effective than similar doses of EHDP in decreasing bone resorption. Osteoclast population parameters were increased with all doses of both Cl2MDP and EHDP with Cl2MDP having a greater effect than similar doses of EHDP. Decreases in the proliferation of the osteoprogenitor pool parallel the decreases in osteoblasts and bone formation parameters. These decreases in osteoprogenitor pool proliferation do not account for the increases with diphosphonates in osteoclast population parameters.
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  • 6
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    Calcified tissue international 23 (1977), S. 283-289 
    ISSN: 1432-0827
    Keywords: Bone ; Resorption ; Osteoclast ; Quantitation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The stimulation of bone resorption, assessed by the release of45Ca from prelabeled bones, was associated with an increase in number of osteoclasts per bone section in parathyroid hormone (PTH)-treated bones, but not in lipopolysaccharide (LPS)-treated bones. By contrast the number of nuclei per osteoclast increased following LPS treatment, but was not affected by PTH. LPS-treated bones had more multinucleated cells, some having as many as 27 nuclei per osteoclast. More osteoclasts were adjacent to the bone collar in bones treated with LPS or PTH than in control bones. In LPS-treated bones this area also contained the largest osteoclasts, as determined by the greatest number of nuclei per osteoclast. The results suggest that LPS and PTH stimulate osteoclastic resorption by different mechanisms.
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  • 7
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    Calcified tissue international 23 (1977), S. 303-305 
    ISSN: 1432-0827
    Keywords: Albumin ; Bone ; Hydroxyapatite ; Plasma protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The extractability of125I-labelled plasma albumin from bone pieces and from powdered bone has been compared after both in vivo and in vitro incorporation. The results show that albumin is more readily extracted from bone pieces than from bone powder which implies that tissue disruption exposes additional protein adsorption sites. It is suggested that incorporation of plasma albumin into calcified matrix during bone formation occurs mainly as a result of its strong interaction with bone mineral.
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  • 8
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    Calcified tissue international 23 (1977), S. 161-170 
    ISSN: 1432-0827
    Keywords: Bone ; Growth ; Rat ; Remodeling ; Tetracyclines
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Cortical bone remodeling along the femur diaphysis was determined in normal female rats (Sprague-Dawley) with the tetracycline technique. Three segments on the cortical bone circumference (the anterolateral, the medial, and the posterior) were found to be most suitable for the study of the remodeling process. Oxytetracycline was administered at age 60 and 75 days, and groups of animals were killed at age 75, 85, 95, and 105 days. The accumulated endosteal growth during age 60 to 75 days in the anterolateral segment was found to increase uniformly in the distal direction along the femur diaphysis. A method is described where this accumulated endosteal growth is used. This method eliminates the use of calipers to determine the section level and makes it possible to study comparable sections even after varying periods of time. The proximal part of the diaphysis showed periosteal apposition in all three segments. The periosteal apposition turned into resorption in the distal part of the diaphysis in the anterolateral and medial segments, whereas the periosteal appsition increased in the posterior segment. The endosteal growth increased in the distal direction in the anterolateral and medial segments. Irregular OTC bands made measurements of endosteal remodeling in the posterior segment impossible. The cortical width decreased in the distal direction along the femoral shaft. Comparison between the different age groups is described and also the relation between the accumulated endosteal growth and the diameter of the medullary cavity.
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  • 9
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    Calcified tissue international 27 (1979), S. 177-185 
    ISSN: 1432-0827
    Keywords: Calcification ; Bone ; Glycoprotein ; Golgi ; Osteoblasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The elaboration of bone matrix glycoprotein by osteoblasts of alveolar bone was investigated by radioautography after the intravenous injection of3H-fucose into young rats. At selected times after injection, animals were sacrificed by intracardiac perfusion and demineralized specimens were prepared for light and electron microscope radioautography. At 5 and 10 min after injection, when the blood fucose level was high, silver grains were restricted to the spheroidal and cylindrical saccules of the Golgi apparatus. At 20 min membrane-limited secretory granules were also labeled. By 35 min, the blood fucose level had dropped and silver grains were detected over the apical cortical cytoplasm, in association with secretory granules located therein. Some grains were present over osteoblast processes and the adjacent prebone matrix. By 4 h most of the silver grains had left the cell. At that time they were observed over prebone, adjacent to osteoblast processes, as well as over the prebone-bone junction where a distinct band of label was noted. In demineralized preparations an electron-dense granular material was present at the prebone-bone junction in association with collagen fibrils. These findings provide evidence that osteoblasts in alveolar bone synthesize fucose-containing glycoprotein and indicate that the addition of3H-fucose occurs in the Golgi apparatus. The glycoprotein passes to the apical cortical cytoplasm by way of membrane-limited secretory granules, is exteriorized, and accumulates at the site where prebone transforms into bone (the prebone-bone junction). Since this is also the site of the calcification front, the deposition of labeled glycoprotein may be related to the deposition of bone mineral.
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  • 10
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    Calcified tissue international 23 (1977), S. 1-5 
    ISSN: 1432-0827
    Keywords: Water ; Bone ; Homeostasis ; Structure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary A short review is given of the water in bone. Various analyses of bone water content are discussed, and its possible location is considered in relation to the behaviour of water in isolated components of bone. Some of the difficulties encountered in examining such microscopic phenomena as water structure in a heterogeneous system such as bone are also discussed.
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  • 11
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    Calcified tissue international 24 (1977), S. 41-46 
    ISSN: 1432-0827
    Keywords: Lead salt ; Bone ; Mobilization ; Hypercalcemia ; Hyperphosphatemia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The present study is an investigation of the mechanism of hypercalcemia and hyperphosphatemia induced by the intravenous injection of lead acetate (Pb-Ac). A total of 118 male rats were injected with 30 mg/kg of Pb-Ac, or with 16.5 mg/kg of sodium acetate as the control. The levels of serum calcium, phosphorus and lead were then determined at various time periods after the injections. Serum calcium and phosphorus levels increased with time after Pb-Ac injection and the maximum values of calcium (17 mg%) were found after 1 h and of phosphorus (13.5 mg%) after 30 min. Both calcium and phosphorus levels reverted to the normal range after 12 h. The maximum net rates of increase of calcium and phosphorus were found immediately after Pb-Ac injection. At that time, deposition of lead at the calcifying sites of bone and incisor dentin was demonstrated by a histochemical examination. In other experiments the changes in the calcium and phosphorus contents in the medium after shaking bone powder in serum with Pb-Ac in an in vitro system were studied. It was confirmed that the calcium and phosphorus were displaced from the bone mineral, the extent of the displacement being correlated with the concentration of the Pb-Ac added to the medium, and that these displacements were very rapid reactions. These results suggest that hypercalcemia and hyperphosphatemia following Pb-Ac injection results from a direct action of lead on the bone mineral.
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  • 12
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    Calcified tissue international 27 (1979), S. 95-99 
    ISSN: 1432-0827
    Keywords: Bone ; Fluoride ; Analysis ; Food ; Age
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Recently published bone fluoride values from Iowa are very high compared to earlier reports, suggesting an increase in fluoride intake. Reanalysis of the Iowa specimens shows levels one-fourth those reported by the Iowa laboratory indicating an error in the original report. Seventeen bone specimens, collected from long-term residents of Rochester, New York, drinking 1 ppm F− water, had a mean value of 2085±270 ppm F− on an ashed-weight basis. This value is not significantly different from that predicted by the data of Zipkin et al. in 1958. These data, therefore, do not support the contention that there has been an increase in fluoride intake.
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  • 13
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    Calcified tissue international 24 (1977), S. 209-214 
    ISSN: 1432-0827
    Keywords: Parathyroid hormone ; Plasma calcium ; Radiocalcium ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The net rate of loss of45Ca from plasma was monitored in normal, thyroparathyroidectomized (TPTX), and nephrectomized rats. Also studied was the effect of a 24 h intravenous infusion of PTH in TPTX rats. The45Ca was injected after an overnight fast and 1 to 4 h prior to the time the first blood sample was obtained. In normal rats, the net rate of loss of45Ca from plasma was slower than in TPTX rats for at least 12 h after radionuclide injection. Nephrectomy did not affect this difference between control and TPTX rats until 8 h after45Ca injection or about 18 h after nephrectomy. At this time a reversal occurred and the net rate of plasma45Ca loss in nephrectomized rats with parathyroids became faster than in TPTX nephrectomized rats. When PTH was infused at a constant rate (3 mU/g body weight/h) plasma calcium concentrations rose gradually for approximately 12 h after which a constant plasma calcium level was maintained with continued PTH infusion. During the period when plasma calcium concentrations were rising, the net rate of loss of45Ca from plasma was slower than in TPTX controls infused only with saline. However, when plasma levels stabilized, plasma45Ca disappearance rates changed such that the rate loss of45Ca from plasma became faster in PTH-infused than in TPTX controls. It is concluded that PTH increases calcium efflux from the same compartment in bone which receives calcium from plasma, thereby returning some recently deposited45Ca to plasma. In the non-equilibrated state, this returns additional45Ca to plasma causing anet decrease in the rate of loss of45Ca from plasma. However, in the equilibrated condition, when both calcium efflux from bone and influx into bone are in balance but at higher flux rates, PTH stimulates the diffusion of45Ca into other bone compartments.
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  • 14
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    Calcified tissue international 28 (1979), S. 103-105 
    ISSN: 1432-0827
    Keywords: Bone ; Photodensitometry, X-ray ; Radiography ; Spectrophotometry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary A conventional spectrophotometer was used to do radiographic photodensitometry of bone mass. An experiment to produce osteoporosis in guinea pigs required quantification of changes in bone mass. Lack of a densitometer necessitated finding another method for quantification. The method developed involved using industrial film in a closed X-ray system. Optical density of X-rays was then read in a spectrophotometer at nine femur locations. Highly significant correlations between optical density and femur calcium content and femur dry weight per millimeter were obtained. Histological sections from an animal with low optical density measurements confirmed bone loss by showing decreased cortical width and loss of trabecular bone. In addition to using readily available equipment, this method offers the advantage of being able to detect bone loss in specific areas of the bones. In addition, bones are left intact and can be used for other purposes.
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  • 15
    ISSN: 1432-0827
    Keywords: ESR ; Bone ; Enamel ; CO3-Apatite ; Paramagnetism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Carbonate-containing hydroxyapatite, enamel, and bone were irradiated by an X-ray and investigated between 77° and 350°K by means of electron spin resonance (ESR) spectroscopy. The ESR spectrum of enamel irradiated at 77°K in vacuum and observed at the same temperature was almost the same as that of the carbonate-containing hydroxyapatite. The temperature dependence of signal intensities confirms a spin-energy exchange between the mineral and organic constituents in bone, but in enamel no or very little spinenergy exchange between the mineral and organic constituents. Considerable similarity among the ESR spectra of enamel, bone, and carbonate-containing apatite was obtained after X-ray irradiation in air at 300°K with both an X-band and a Q-band ESR spectrometer. The Q-band spectrum can be interpreted in terms of two paramagnetic species. One is identified as a CO 3 3− anion radical which has an axial symmetry withg factors of 2.0029 and 1.9972. The other species is found to be centered atg=2.0019.
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  • 16
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    Calcified tissue international 28 (1979), S. 33-36 
    ISSN: 1432-0827
    Keywords: Infrared, visible light ; Bone ; Collagen ; Apatite
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Photovoltaic effect (infrared and visible light) is observed in bone and its two major components, collagen and apatite, at room temperature. A dimunition in the magnitude of photovoltage is observed after exposure to ultraviolet light in all the cases. The drift mobility of the charge carriers is obtained by measuring I versus V relationships in sandwich samples and relating them to the permitivity of the medium. Lifetime of the injected carriers is measured in the usual way. The data are consistent with the hypothesis that the effects are due to protonic conduction phenomena.
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  • 17
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    Calcified tissue international 26 (1978), S. 47-49 
    ISSN: 1432-0827
    Keywords: Matrix vesicles ; Calcospherites ; Mineralization ; Bone ; Dentin ; Cartilage ; SEM
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Mineral particle cluster, corresponding to calcospherites or matrix vesicle-initiated clusters in calcifying cartilage and dentin and to collagen bundle-related mineralization in lamellar bone, have been isolated from NaOCl solutions used to dissolve the organic matrix in preparation of anorganic mineralizing fronts for scanning electron microscopy.
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  • 18
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    Calcified tissue international 26 (1978), S. 81-84 
    ISSN: 1432-0827
    Keywords: Osteoclasts ; Bone ; Resorption
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Osteoclastic bone resorption involves the solubilization of the mineral salts and the degradation of noncollagen bone matrix and collagen fibrils. As no recognizable collagen fibrils have ever been reported within cytoplasmic vacuoles in osteoclasts, it is generally assumed that the collagen fibrils are digested extracellularly in the resorption zone. The extent to which lysis occurs extracellularly and whether or not the osteoclasts phagocytose the degradation products remain to be established. In the present communication, a hypothesis is presented suggesting the possibility that osteoclastic resorption of bone involves the participation of two different cell types. According to this hypothesis, osteoclastic bone resorption is initiated by osteoclasts that demineralize areas of bone and degrade noncollagen bone matrix. After the osteoclasts have moved away or become partially detached from the demineralized site, the exposed collagen fibrils are phagocytosed by mononuclear, fibroblast-like or monocyte-derived cells.
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  • 19
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    Calcified tissue international 26 (1978), S. 155-161 
    ISSN: 1432-0827
    Keywords: Plasma protein ; Glycoprotein ; Bone ; Glucosamine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Plasmaα 2HS-glycoprotein is specifically accumulated in calcified tissues. In the present studies this glycoprotein was isolated from plasma and after iodination with iodine-125 was injected intravenously into young rabbits. The tissue distribution and plasma disappearance rate of this radioactively labeled material were determined. Of the various tissues studied, bone showed the greatest retention of labeled glycoprotein expressed as percentage of the injected dose per gram tissue relative to the plasma content. The rate of loss of iodinatedα 2HS-glycoprotein from plasma was similar to that ofα 2HS-glycoprotein labeled endogenously by using14C-glucosamine or3H-glucosamine. The uptake of exogenously labeled3I-α 2HS-glycoprotein into bone tissue expressed as a percentage of the injected dose was similar to that of endogenously labeled14C-α 2HS-glycoprotein. These results suggest that the125I-labeled material can be used to study further the metabolism ofα 2HS-glycoprotein by bone tissue.
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  • 20
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    Calcified tissue international 26 (1978), S. 209-213 
    ISSN: 1432-0827
    Keywords: Mithramycin ; Bone ; β-Glucuronidase ; KT medium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Mithramycin suppresses bone resorption. Its effect on the synthesis and release of β-glucuronidase (a referent for lysosomal enzymes) in mouse calvarial explants was studied in an in vitro culture system. A newly described medium (designated as KT medium) was introduced in this specific study. Mithramycin initially inhibited the release of β-glucuronidase into the medium and resulted in an ultimate accumulation of this enzyme in the bone. These results suggest that inhibition of bone resorption by mithramycin may be attributed to interference in release of lysosomal enzymes from bone cells.
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  • 21
    ISSN: 1432-0827
    Keywords: Vitamin D ; Bone ; Osteoblasts ; Receptors ; 1,25-dihydroxyvitamin D3
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Thaw-mount autoradiographic studies after injection of3H-1,25-D3 were conducted on 18-and 20-day-old rat fetuses. In maxillary bones, ribs, and tibia, nuclear concentration of radioactivity was found in osteoprogenitor cells and osteoblasts. Osteocytes and chondrocytes in epiphyseal plates were either unlabeled or weakly labeled. In competition experiments, nuclear concentration of radioactivity was blocked by the injection of a high dose of nonradioactive 1,25-D3 prior to the administration of the labeled hormone, but not by a similar dose of nonradioactive 25-D3. The results are interpreted as indicating that osteoprogenitor cells and osteoblasts are target cells for the direct action of 1,25-D3 on fetal bone.
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  • 22
    ISSN: 1432-0827
    Keywords: Vitamin D ; Vitamin D deficiency ; Bone ; Cartilage ; Bone development
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The role of vitamin D in early skeletal development was studied by measuring serum calcium and phosphorus, osseous tissue quantity and mineralization, and endochondral bone elongation in rat fetuses and pups from vitamin D-replete and vitamin D-deficient mothers. At the 20th day of pregnancy there was a slight, yet significant, increase in the amount of osteoid on trabecular bone surfaces in fetuses from vitamin D-deficient mothers. The fetal bones otherwise appeared normal in spite of severe skeletal changes in the vitamin D-deficient mothers. After parturition, the importance of vitamin D in skeletal development becomes progressively more obvious. Serum calcium levels were slightly, yet significantly, lower in vitamin D-deficient than in vitamin D-replete pups and these levels continued to fall in the vitamin D-deficient pups through lactation and after weaning. At 3 days postpartum, there was a small, yet significant, increase in the amount of osteoid on bone surfaces of the vitamin D-deficient pups. The relative amounts of osteoid in the vitamin D-deficient pups continued to increase through lactation and after weaning when compared with vitamin D-replete pups. By the 14th day of lactation and at later periods, there were significant reductions in metaphyseal mineralized tissues in the vitamin D-deficient pups when compared with the vitamin D-replete pups. At weaning and after weaning, there were substantial increases in growth plate thickness and decreases in longitudinal bone growth in the vitamin D-deficient pups when compared with the vitamin D-replete pups. The results from this study indicate that vitamin D does not appear to play a major role in fetal skeletal development. However, after birth, vitamin D becomes progressively more important with age for normal bone development, mineralization, and endochondral growth.
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  • 23
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    Calcified tissue international 35 (1983), S. 481-485 
    ISSN: 1432-0827
    Keywords: Chemotaxis ; Bone ; Osteoblasts ; Bone proteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary When demineralized bone matrix powder is implanted subcutaneously in the rat, the early responses involve the appearance and proliferation of mesenchymal cells at the site of implantation, followed by cartilage and bone formation. The ability of cells to migrate to the implant suggests that chemotaxis may be a critical event in this process. Therefore, using the modified Boyden chamber assay, we tested extracts of demineralized bone matrix for chemotactic activity. We have identified and partially purified, on molecular sieve chromatography, a heat labile and trypsin-sensitive protein (Mr=60,000–70,000) that is a potent chemoattractant for mouse calvaria, osteoblast-like cells (MMB-1), but not for monocytes (putative osteoclast precursors). These findings suggest that chemotactic protein(s) have a significant role in the recruitment of osteoprogenitor cells to a site of bone repair.
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  • 24
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    Calcified tissue international 35 (1983), S. 762-766 
    ISSN: 1432-0827
    Keywords: Enamel ; Maturation cycles ; Tetracycline ; Dentine ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Uptake of tetracycline by enamel in the short-term was studied at an advanced stage of crown formation and after completion of crown formation in deciduous molars in the cat. Both secretory phase enamel and bands of postsecretory, maturation phase enamel labeled rapidly. The pattern of labeling mimicked that seen in the continuously growing, rootless incisor teeth of the rat, with narrow doublets fusing to form narrow bands with wide unlabeled intervals in the short term. This is a physiological demonstration which indicates that cyclical activity and changes may occurin vivo during the maturation phase of amelogenesis in rooted teeth. It is also noted that dentine did not, and that some circumscribed patches of bone did label in the same animals in the same time interval. Short-term tetracycline labels are lost following conventional histological processing, but are retained after freeze-drying or air-drying.
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    Calcified tissue international 36 (1984), S. S25 
    ISSN: 1432-0827
    Keywords: Remodeling ; Bone ; Stress
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    Topics: Biology , Medicine , Physics
    Notes: Summary It has long been known that the stress history of bone tissue influences its structure; however, the nature of this relationship remains largely uncharacterized. The objective of this work was to induce a quantifiable change in the stress history ofin vivo bone tissue and examine subsequent changes in structural and material properties that might occur. Continuous compressive loads were applied to the diaphysis of adult mongrel dogs for 2 months. The loads, ranging from 12–130 N, were superposed on the normal activity of the animals by implanting spring loading devices on the diaphysis of the femur. After the animals were sacrificed, mid-diaphysial specimens were subjected to compression testing to determine a structural bulk stiffness. The cross-sectional areas of original bone tissue and new bone deposition were then determined. The ash weights of selected specimens were also determined. The results indicate that a positive correlation between the increase in cross-sectional area and the superposed stress does exist. The new bone apposition was found almost entirely on the periosteal surface. Very little evidence of internal remodeling or endosteal movement was observed. The new tissue was found to have a lower ash weight and appeared to have a disorganized microstructure. Mechanical testing also suggests that the newly deposited tissue is far less stiff than the mature original bone.
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  • 26
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    Calcified tissue international 34 (1982), S. 439-448 
    ISSN: 1432-0827
    Keywords: Bone ; Histomorphometry ; Osteocytes ; Bone dynamics ; Histology
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    Topics: Biology , Medicine , Physics
    Notes: Summary A new semiautomatic technique combining advantages of the manual and fully automatic methods is described for obtaining quantitative static and dynamic histologic data of bone. The hardware consists of a photomicroscope, digitizing platen, digitizer, plotter/printer, floppy disc drive, and computer. The microscope is equipped with a drawing tube through which the image of the digitizing platen is projected over the optical field. The investigator selects and traces all histologic structures to be measured by moving a cursor on the digitizing platen which is visible by its projection over the histologic field. The results on accuracy and static and dynamic precision of this method show that static and dynamic parameters of bone are obtained with a degree of error (〈20%) well within the acceptable range for biologic measurements. Comparison of this method with the grid technique according to Merz and Schenck showed that for almost all micromorphometric parameters comparable absolute data are obtained. Due to the higher precision of our method, however, the number of optical fields evaluated in obtaining these comparable data could be reduced to 25% of the number of fields evaluated by the Merz and Schenck technique. The time requirements for quantitative evaluation of a histologic slide of bone by our technique are 40–50 min; 20–25 min is needed for quantitative evaluation of osteocytes.
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    Calcified tissue international 34 (1982), S. 449-455 
    ISSN: 1432-0827
    Keywords: Bone ; Histomorphometry ; Normal values ; Bone biopsies ; Sampling error
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    Topics: Biology , Medicine , Physics
    Notes: Summary Quantitative bone histology was done in undecalcified sections of iliac crest bone specimens obtained from 84 normal American individuals. Samples were obtained within 12 h after death in a vertical and horizontal manner from both the right and left iliac crests. In addition to the determination of normal values of micromorphometric parameters of bone in these healthy American subjects, the following studies were carried out: (a) comparison of variance of micromorphometric parameters of bone obtained from the right versus left iliac bone (40 pairs), (b) comparison of micromorphometric parameters of bone obtained in a vertical versus horizontal manner (12 pairs), (c) evaluation of variance with increasing distance from the compact zone in bone samples obtained in a vertical manner (44 pairs), (d) analysis of variation between bone samples obtained more anteriorly versus posteriorly along the iliac crest (N=40), (e) comparison of differences in micromorphometric parameters obtained from age-matched men versus premenopausal women (N=12), and (f) plotting of histograms for assessment of distribution of micromorphometric parameters. The results show that histomorphometric data of bone cannot be easily compared when different techniques are employed for obtaining bone samples. Sampling variations are kept smaller when bone specimens are obtained in a vertical manner. Anterior/posterior variation does not cause major sampling error. If ranges of variation are taken into account, quantitative bone histology is a valuable tool for assessment of bone structure and bone cells.
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    Calcified tissue international 35 (1983), S. 148-152 
    ISSN: 1432-0827
    Keywords: Bone ; Computed tomography ; Densitometry ; Osteoporosis ; Osteomalacia
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    Topics: Biology , Medicine , Physics
    Notes: Summary The linear attenuation coefficient (μ in cm1) of trabecular bone was modeled for different conditions of bone and marrow composition in order to assess their influence on computed tomography (CT) quantitation. A large relative change (10% of TBV at 15% TBV) of bone concentration resulted in small changes of μ: 2.3% at 60 keV, 3.4% at 44 keV, 5.2% at 29 keV. Relative changes of trabecular bone volume (TBV) on the order of 3% could be detectedin vivo by CT were it not for errors of relocation and for compositional influences on accuracy. The μ (and density) depended critically not only on amounts of bone substance and marrow but on their compositions. Normal variation in the composition of bone substance produced an uncertainty in μ equivalent to 0.5 to 1% TBV. Increases of yellow marrow produced a decrease of μ which could be mistaken for a decrease of bone concentration. The biological variation (90% confidence limit) of marrow composition gives an uncertainty at 15% TBV of about 2.4% TBV at 60 keV, 1.7% at 44 keV, and 1.3% at 29 keV. These correspond to relative uncertainities of 16, 11, and 9% respectively. These factors help explain the large accuracy errors (30%) observed in all studies of trabecular bone where single-energy CT was used. Marrow composition also can affect precision of bone measurement. Systematic shifts of red and yellow marrow could mask biological changes such as those occurring with aging or treatment.
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    Journal of molecular evolution 21 (1984), S. 19-32 
    ISSN: 1432-1432
    Keywords: Leghemoglobin ; Gene duplication ; Gene linkage ; Concerted evolution ; Nitrogen fixation ; Soybean
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    Topics: Biology
    Notes: Summary We have analyzed the sequences of soybean leghemoglobin genes as an initial step toward understanding their mode of evolution. Alignment of the sequences of plant globin genes with those of animals reveals that (i) based on the proportion of nucleotide substitutions that have occurred at the first, second, and third codon positions, the time of divergence of plant and animal globin gene families appears to be extremely remote (between 900 million and 1.4 billion years ago, if one assumes constancy of evolutionary rate in both the plant and animal lineages) and (ii) in addition to the normal regulatory sequences on the 5′ end, an approximately 30-base-pair sequence, specific to globin genes, that surrounds the cap site is conserved between the plant and animal globin genes. Comparison of the leghemoglobin sequences with one another shows that (i) the relative amount of sequence divergence in various coding and noncoding regions is roughly similar to that found for animal globin genes and (ii) as in animal globin genes, the positions of insertions and deletions in the intervening sequences often coincide with the locations of direct repeats. Thus, the mode of evolution of the plant globin genes appears to resemble, in many ways, that of their animal counterparts. We contrast the overall intergenic organization of the plant globin genes with that of animal genes, and discuss the possibility of the concerted evolution of the leghemoglobin genes.
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    Calcified tissue international 33 (1981), S. 111-117 
    ISSN: 1432-0827
    Keywords: Calcium-deficient hydroxyapatite ; Amorphous calcium phosphate ; Bone ; Radial distribution function ; Carbonate apatite
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    Topics: Biology , Medicine , Physics
    Notes: Summary When amorphous calcium phosphate (ACP) was transformed to crystalline hydroxyapatite (HA) in a series of aqueous slurry concentrations ranging from low to high, the higher slurry concentrations produced more Ca-deficient HA as measured by Ca/P ratio and heat-produced pyrophosphate. We feel that the excess solution phosphate produced in the higher slurry transformations results in lower Ca/P ratio HA. It has been suggested that an ACP is the precursor to bone apatite. Regulation of the in vivo ACP slurry concentration could then control the stoichiometry and, therefore, the metabolic activity of bone apatite. X-ray radial distribution function (RDF) analyses showed that CO 3 2− substitution in HA creates far greater structural distortions than do Ca deficiencies. The latter, however, do produce small, but observable, structural distortions when compared to stoichiometric HA. It now seems clear that the RDF of bone apatite can be modeled by a synthetic, Ca-deficient, CO 3 2− -containing HA.
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  • 31
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    Keywords: Phosphoprotein ; Bone ; 31P-NMR ; Phosphoserine ; Phosphothreonine
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    Topics: Biology , Medicine , Physics
    Notes: Summary Fractionation of the EDTA-soluble, noncollagenous proteins of the organic matrix of chicken bone by Sephadex G-100 molecular sieving has revealed that the majority of the organic phosphorus is present in two fractions, from one of which a homogeneous phosphoprotein has been isolated. The purified phosphoprotein has an apparent molecular weight of 12,000 and contains bothO-phosphoserine andO-phosphothreonine.31P-NMR spectroscopy demonstrates that all of the organic phosphorus exists in the form of phosphomonoesters which have an average pK2 of 6.8. The phosphoprotein is highly acidic due to its high content of dicarboxylic acids in addition to the presence of organic phosphorus. The characteristic amino acid composition of the phosphoprotein establishes its noncollagenous nature and highlights the differences among bone, dentin, and enamel phosphoproteins. The absence ofγ-carboxyglutamic acid distinguishes it from osteocalcin, the noncollagenousγ-carboxyglutamic acid-containing peptide of bone matrix.
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    Calcified tissue international 33 (1981), S. 455-457 
    ISSN: 1432-0827
    Keywords: Bone ; Osteoporosis ; Diabetes mellitus
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    Topics: Biology , Medicine , Physics
    Notes: Summary The bone mineral status of fifty-one children with diabetes mellitus was studied by single photon absorptiometry. The mean bone mineral content was 13% below values predicted by age, sex, height, and weight. Those children whose diabetes was one year or less in duration were as osteopenic as those whose diabetes was of longer duration. The demineralized children received a higher daily insulin dose than the others. No association was noted between the degree of skeletal demineralization and sex, statural growth, renal function, and serum calcium and phosphorus. No significant changes in bone mineral content were noted longitudinally.
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    Calcified tissue international 23 (1977), S. 185-188 
    ISSN: 1432-0827
    Keywords: Matrix vesicles ; Bone ; Ultracrytomy
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    Topics: Biology , Medicine , Physics
    Notes: Summary Matrix vesicles were observed in femurs of 8-day-old chick embryos prepared by ultracryotomy. Some of the sections were subjected to ultramicroincineration. The unfixed tissues never came into contact with solutions, and thereby artifacts due to dissolution, redistribution, or rearrangement of the mineral constituents were avoided. In the osteoid, electron dense objects with the size and appearance of matrix vesicles were seen, although limiting membranes were not visible. After ultramicroincineration the vesicles were observed to contain small crystals and a less dense amorphous mineral material which may be the precursor of bone mineral. In addition, a ring of ash enclosed the crystalline and amorphous mineral and appeared to occupy the position of the vesicle membrane as seen in conventionally prepared material.
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    Calcified tissue international 24 (1977), S. 93-98 
    ISSN: 1432-0827
    Keywords: Magnesium ; Bone ; Mineral ; Chick embryo ; Organ culture
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    Topics: Biology , Medicine , Physics
    Notes: Summary Using bone organ culture techniques, three concentrations of magnesium were evaluated for their effects on the mineral mass of embryonic chick tibiae incubated for 3 days with or without a metabolic inhibitor (1 mM iodoacetic acid) added to the media. Varying the medium Mg level from 0.3 to 3.0 mM had little effect on the net increase in mineral in live bones (without the inhibitor), but in dead tibiae (with the inhibitor) there was a marked net decrease in mineral deposition with increasing Mg concentration. Bone Mg mass varied directly with the level of Mg in the medium regardless of whether or not the tibiae were inhibited. Responsiveness to parathyroid hormone (PTH) with regard to mineral loss and increased lactate production was not affected by Mg even in tibiae incubated in Mg-free media. However, PTH-treated bones cultured in Mg-free media had a significantly elevated Mg mass, suggesting that the hormone may have a Mg-conserving effect on bone during severe Mg deficiency. The findings of this study support the hypothesis that bone cells regulate the level of Mg to which newly forming mineral is exposed and that PTH-responsiveness in embryonic chick tibiae is not dependent on the ambient Mg concentration.
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    Calcified tissue international 22 (1977), S. 9-17 
    ISSN: 1432-0827
    Keywords: Bone ; Fluoride ; Calcium ; Phosphorus ; Mechanical Strength
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    Topics: Biology , Medicine , Physics
    Notes: Abstract Four groups of weanling rats were fed for 2 weeks on a diet sufficient or insufficient in calcium and/or phosphorus. Each group was divided into four subgroups which were offered distilled water supplemented with 0, 50, 75, or 150 ppm fluoride. High levels of fluoride in drinking water inhibited weight gain. This inhibition was less in rats deficient in phosphorus than when normal-phosphorus diets were offered. At a low level, fluoride was without any effect on bone ash, thickness of femoral cortical bone, and mechanical strength, as measured by maximal load, ultimate stress to breaking, and limit of elasticity. Modulus of elasticity was decreased. At higher levels fluoride tended to decrease most of these parameters, except in rats deprived of both calcium and phosphorus. The effect of fluoride was modified by lack of dietary calcium and/or phosphorus and appeared to be weaker in rats deficient in these nutrients. Lack of dietary calcium and/or phosphorus decreased bone strength more than did fluoride content of water and of bone mineral. Concentration of bone ash and thickness of femoral cortical bone were closely correlated with parameters of mechanical strength.
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  • 36
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    Keywords: Parathyroid hormone ; Plasma calcium ; Radiocalcium ; Bone
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    Notes: Abstract Acute changes in plasma calcium and45Ca were studied in young adult male thyroparathyroidectomized (TPTX) rats injected with moderate doses of parathyroid hormone (PTH). For plasma calcium changes, comparison was made between rats fasted or fed prior to PTH injection. For plasma45Ca changes, the effect of the time of administration of the radionuclide was also studied; this included rats injected with PTH 1 h after radionuclide (“1 h45Ca”), 18 h later (“18 h45Ca”) and more than 6 days later (“6 day45Ca”). The results can be summarized as follows: (1) Plasma calcium changes were greater when PTH was injected into “fed” rather than into “fasted” rats. (2) PTH always produced a relative increase (compared to controls tested concurrently) in plasma45Ca concentrations. This increase was the same in the “1 h45Ca” and the “18 h45Ca” groups. (3) Plasma45Ca rose at least temporarily following PTH injection in the “18 h45Ca” group. (4) The45Ca rise following PTH was always greater in “fed” than in “fasted” groups. (5) Plasma45Ca specific activities (S.A.) tended to rise in the “6 day45Ca” group and to fall in the “18 h45Ca” group, following PTH injection. However, the45Ca S.A. was always higher in fed than fasted groups. (6) In a few experiments in which32P was injected with45Ca, specific activity changes in plasma45Ca following PTH injection werenot accompanied by similar changes in32P specific activity. These results could not be adequately explained by PTH effects on bone resorption, but the data supported the postulate that PTH controls plasma calcium concentrations by increasing transport of calcium through the osteocyte-lining cell (osteoblast) bone cell complex from the bone fluid compartment to the ECF.
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    Calcified tissue international 22 (1977), S. 275-284 
    ISSN: 1432-0827
    Keywords: Bone ; Calcium ; Collagen ; Dentine ; Phosphoprotein
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    Topics: Biology , Medicine , Physics
    Notes: Abstract Calcium-binding and biochemical studies have been applied to characterize the state of the carboxylate and protein-bound phosphate groups in the EDTA-demineralized matrices of rat bone and dentine. The organic phosphate and carboxylate content of demineralized bone is virtually identical to that of purified steer skin collagen whereas demineralized dentine has a significantly higher phosphate and carboxylate content, presumably due to the presence of an acidic non-collagenous phosphoprotein. Two classes of calcium-binding sites can be detected in demineralized bone, demineralized dentine, and purified, reconstituted collagen. The number of strong calcium-binding sites correlates with the number of protein-bound phosphate groups. Depending on the preparative procedure, seven to nine such sites (per collagen molecule) are present in dentine, and one to two in the purified reconstituted collagen and in bone. The binding constant for the dentinal sites (1.1×104 M−1), however, is 20 times greater than that for bone or reconstituted collagen fibrils from skin. We tentatively conclude that the strong calcium-binding site in bone and reconstituted collagen is of the form protein-PO 4 − Ca++ whereas in dentine it is of the form the weak binding sites in bone and dentine are of the form protein-COO-Ca++; and that approximately 160 of the 217 carboxylate groups of the collagen molecules of dentine or bone are present as electrostatic linkages of the form protein-COO−+H3N-protein.
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    Calcified tissue international 22 (1977), S. 315-327 
    ISSN: 1432-0827
    Keywords: Bone ; Bioelectricity ; Strain ; Locomotion ; Osteoporosis
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    Topics: Biology , Medicine , Physics
    Notes: Abstract Using wet dead specimens it was possible to show that the size and form of strain-related electrical potentials on the surface of sheep radii were related both to the amount of strain, and the strain rate, over the range of these recorded during locomotion. Using the same electrode and amplifier system in vivo changes in surface strain and surface charge were recorded from the radius of three sheep during locomotion. During slow locomotion electrical changes were negligible. At medium speed they were most variable, and profoundly influenced by small alterations in the timing and pattern of strain change. When locomotion was fast the electrical waveform reflected fairly faithfully the changes in strain on the bone surface.
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    Calcified tissue international 22 (1977), S. 329-331 
    ISSN: 1432-0827
    Keywords: Bone ; Fracture ; Mineral
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    Topics: Biology , Medicine , Physics
    Notes: Abstract The bone mineral content was measured by gamma absorptiometry in patients who had sustained fractures of the upper limb. Measurements were taken on the shafts of the forearm and in the trabecular bone immediately proximal to the wrist. Fractures of the surgical neck of the humerus and fractures of the shaft of the humerus did not significantly influence the bone mineral content of the ipsilateral forearm. Fracture of the radius and the ulna, however, caused a 15% loss of the bone mineral content in the trabecular bone proximal to the wrist, whereas the mineral content on the shafts of the fractured forearms increased or decreased depending on the fracture site. The findings suggest that measurements of bone mineral content of the forearm are not necessarily invalidated if there has been a fracture of the same limb. Only if the forearm itself is fractured should measurements on this site be avoided.
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    Calcified tissue international 25 (1978), S. 37-43 
    ISSN: 1432-0827
    Keywords: ESR ; Radiation free radicals ; CO3-Apatites ; Bone
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    Topics: Biology , Medicine , Physics
    Notes: Summary Exposure of bone mineral to X-rays generates free radicals. These are usually very labile, but can be stabilized at liquid nitrogen temperatures for study by electron spin resonance spectroscopy. The free radicals thus detected in the present study included one with resonances arising from an electron excess center and 2 species with electron-deficit centers: a phosphate anion radical and a radical associated with carbonate. Each of these radicals seemed to be located chiefly at the mineral surface and was sensitive to the surface environment. Presence of an organic phase, as in whole bone, modified free radical production in a manner that suggests interference with the formation of electron deficit centers. Comparison with other synthetic minerals suggests that precipitated carbonateapatites are good models for bone mineral.
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    Calcified tissue international 27 (1979), S. 33-40 
    ISSN: 1432-0827
    Keywords: Chick embryo ; Bone ; Organ culture ; Scanning electron microscopy
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    Topics: Biology , Medicine , Physics
    Notes: Summary The study describes the ultrastructure of the mineralized portion of chick tibiae from 10 days in ovo to 2 days post-hatch. At 10 days a single mineralized cylinder surrounds the diaphysis. On its outer surface columnar trabeculae join to form ridges parallel to the long axis of the bone. These ridges are covered by another cylinder and form the haversian canals. At 11 days vascular invasion of the marrow cavity occurs and resorption of the endosteal surface begins. This type of periosteal deposition and endosteal resorption is repeated during and subsequent to embryonic development. The mineralized portion of 10-day chick tibiae cultured for 2 days in modified BGJ medium was compared with 10-, 11-, and 12-day tibiae in ovo. Cultured tibiae were similar in length and calcium content to 11-day tibiae in ovo. The form of mineral deposited in ovo and in culture was the same, namely, aggregates of spherical mineral clusters. Differences in culture included the following: (a) few concentric cylinders were deposited as compared with tibiae in ovo; (b) trabeculae were not arranged in rows and ridges in culture; (c) osteocytic lacunae were restricted to bases of trabeculae rather than uniformly distributed as in ovo; and (d) the endosteal surface of tibiae in culture appeared etched.
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  • 42
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    Calcified tissue international 27 (1979), S. 57-64 
    ISSN: 1432-0827
    Keywords: Bone ; Bone formation ; Calcification ; Calcification nodule ; EM
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Osteolathyrism has been used as an experimental model for the study of calcification nodules during the mineralization process. Periosteal exostoses developing in osteolathyrism characteristically have spherical basophilic structures (calcification nodules) in the vicinity of developing bone spicules. In thin sections, the nodules were seen scattered between collagen fibers in the intercellular matrix. Collagen fibers did not appear to be present within the nodules but sometimes were packed just outside them. Matrix vesicles were also present in areas of early mineralization. After EDTA decalcification, the majority of the nodules consisted of a fine granular material surrounded by an electron-dense peripheral zone. The peripheral dense zone was occasionally incomplete in small nodules in areas of early mineralization. An electron-dense central area could be observed in the center of the nodules. Evidence has been presented indicating that the calcification nodules arise from smaller mineralization foci, presumably matrix vesicles. The calcification nodules enlarge to approximately 1.0 µm in size, at which point development is slowed or halted allowing the formation of the peripheral dense zone. Although coalescence of nodules was observed, this was more a random event. The further mineralization of the trabeculae was achieved by the calcification of the collagen fibers. The mineralized trabeculae reflected this pattern of nodular and collagenous calcification. It is suggested that this pattern of calcification is characteristic of rapidly developing woven bone.
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  • 43
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    Calcified tissue international 27 (1979), S. 105-108 
    ISSN: 1432-0827
    Keywords: Bone ; Femur ; Strength ; Density ; Age
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    Topics: Biology , Medicine , Physics
    Notes: Summary In order to ascertain whether the intrinsic strength of human bone changes with age or not, we have determined the ultimate tensile strength and density of strips of femoral cortical bone. These femora were collected from cadavers varying in age from 13 to 97 years. The results show that both density and intrinsic strength of bone increase up to about the fourth decade of life and then decrease with age. However, the rate of decrease of strength is greater than that of density. This indicates that the density of bone is not the sole determining factor of its strength, and that some other factors play an important part.
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  • 44
    ISSN: 1432-0827
    Keywords: Bone ; Rickets ; Hypophosphatemia ; Microradiography ; D-resistance
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    Topics: Biology , Medicine , Physics
    Notes: Summary The perilacunar areas of low mineral density in microradiographs from cortical bone of patients with hypophosphatemic (vitamin D-resistant) rickets are not evenly distributed throughout the bone tissue. Their frequency and distribution were determined in bone from 9 patients with this disease. It was found that the lesion was more frequent in haversian bone than in interstitial bone, and along the inner circumference of growing haversian systems as compared with the outer circumference. These findings indicate that the lesion is the result of retarded mineralization, and that mineralization slowly proceeds in these areas as the bone becomes older. A relatively high frequency of the lesion was also found in osteons with an elliptical cross section along the long axis of the ellipse. The cause of the abundance of the lesion at these sites is not clear, but it is possible to explain the uneven distribution in elliptical osteons by assuming an unequal rate of bone formation in these structures.
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    Calcified tissue international 27 (1979), S. 165-169 
    ISSN: 1432-0827
    Keywords: Oxygen uptake ; Bone ; Oxygen tension ; Carbon dioxide
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    Topics: Biology , Medicine , Physics
    Notes: Summary The present study was carried out to determine the influence of carbon dioxide and oxygen tension on the respiratory activity of bone cells in mouse calvaria in vitro. Five-day-old mouse calvaria were removed aseptically and incubated individually for 1 h at 37° C in a closed reaction chamber containing 1.5 ml of tissue culture medium made up of 60% horse serum in Gey's solution containing 100 unit/ml penicillin and 100µg/ml streptomycin. Before the calvaria were added, the medium in the incubation chamber was equilibrated with 10%, 20%, 30%, or 50% oxygen balanced with nitrogen. The effect of CO2 on oxygen utilization by the calvaria was determined by incubating the calvaria in a medium previously equilibrated with either 50% O2 balanced with N2 or 50% O2 and 5% CO2 balanced with N2. At each oxygen tension, the rate of oxygen utilization by the calvaria was measured polarographically by a Clark oxygen electrode. The results showed that the rate of oxygen uptake of bone increased as the oxygen tension increased and carbon dioxide stimulated significantly the rate of oxygen utilization by the bone cells. In view of the previous reports that both carbon dioxide and oxygen tension are implicated in the process of bone resorption, it is suggested that these two factors may affect bone resorption by influencing the oxygen utilization by bone cells and ultimately controlling their energy metabolism.
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    Calcified tissue international 23 (1977), S. 215-223 
    ISSN: 1432-0827
    Keywords: Amorphous mineral ; Bone ; Electron microscopy ; Ultracryotomy ; Ultramicro-incineration
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    Topics: Biology , Medicine , Physics
    Notes: Summary The fine structure of the extracellular phase of avian medullary bone and embryonic chick femur was examined in thin sections prepared by ultracryotomy and ultramicroincineration. Since contact with solutions was completely avoided, little or no loss or dislocation of mineral constituents could occur. Amorphous bone mineral (ABM) was present in two forms: as 15–30 nm spheres and as a structure-free haze. Removal of all organic material by low temperature ashing left the ABM intact. Crystals were usually associated with the ABM. In newly ossifying regions clusters or nodules of randomly oriented crystals and ABM appeared to coalesce when they reached approximately 1 μm in diameter. In highly calcified regions crystals appeared to be oriented along collagen fibers. ABM did not appear to be associated with collagen. Unmineralized collagen was visible in osteoid after staining with dry OsO4 vapor and it appeared to be diverted around nodules. Structures which resembled matrix vesicles were present. Selected area electron diffraction patterns indicated the presence of hydroxyapatite.
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    Calcified tissue international 28 (1979), S. 37-42 
    ISSN: 1432-0827
    Keywords: X-ray diffraction ; Hydroxyapatite ; Whitlockite ; Fish (Pisces) ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Pyrolyzed scales, fin spines, and bone from the ray-finned bony fishPolypterus (Actinopterygii) showed two mineral phases on X-ray diffraction: hydroxyapatite (HA), Ca5(PO4)3OH, and whitlockite, Ca3(PO4)2. The ratio of HA/whitlockite varied with the structure (scale, spine, bone) within each individual fish. The relative proportions of HA to whitlockite in pyrolyzed samples reflected the Ca/P ratio of the sample. Whitlockite appears after pyrolysis when the Ca/P is lower than 1.67. Among the five fish investigated, for each structure a general trend was noted. The proportion of HA relative to whitlockite increased with size (age) of the fish. Thus the smallest fish, a juvenile, exhibited a low Ca/P mineral in its calcified tissues, whereas the larger fish had progressively more HA and less whitlockite.
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  • 48
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    Calcified tissue international 28 (1979), S. 17-22 
    ISSN: 1432-0827
    Keywords: Anticonvulsant ; Ketogenic diet ; Calcium ; Vitamin D ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Vitamin D and mineral metabolism status was examined in five children maintained chronically on combined ketogenic diet-anticonvulsant drug therapy (KG), and the results compared to those obtained in 18 patients treated with anticonvulsant drugs alone (AD) and 15 normal controls. KG patients exhibited biochemical findings of vitamin D deficiency osteomalacia: decreased serum 25-hydroxyvitamin D (25OHD) and calcium concentrations, elevated serum alkaline phosphatase and parathyroid hormone concentrations, decreased urinary calcium and increased urinary hydroxyproline excretion, and decreased bone mass. Although the KG and AD groups demonstrated similar reductions in serum 25OHD concentration, the KG patients exhibited a significantly greater reduction in bone mass. In response to vitamin D supplementation (5000 IU/day), mean bone mass in the KG group increased by 8.1±0.9% (P〈0.001) over a 12-month period. These results suggest that ketogenic diet and anticonvulsant drug therapy have additive deleterious effects on bone mass and that these effects are partially reversible by vitamin D treatment.
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  • 49
    ISSN: 1432-0827
    Keywords: Bone ; Bone cells ; Cyclic AMP ; Osteoclast activating factor ; Lymphokine
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    Topics: Biology , Medicine , Physics
    Notes: Summary Osteoclast activating factor (OAF) is a lymphokine which may participate in the pathologic destruction of bone observed in a number of disorders. In the current studies, we investigated the action of OAF on cAMP accumulation by bones and isolated bone cells in culture. OAF was shown to stimulate accumulation of cAMP in mouse cranial bones at doses between 1 and 1000 ng/ml. Stimulation of bone resorption was observed in bones treated with the same doses of OAF. In order to investigate the cell types responsible for cAMP responses to OAF, we isolated bone cells and grew them in monolayer culture. The cells were isolated by a variety of techniques which separate bone cells into two types of parathyroid hormone (PTH)-responsive populations: (a) cells derived from the periosteal regions of the bone, which also respond to calcitonin with increases in cAMP; and (b) cells derived from the matrix, which do not respond to calcitonin. OAF caused elevation of cAMP levels in both the periosteum-derived cells and the matrix-derived cells. The magnitudes and time courses of OAF effects in these populations resembled the effects previously reported for PTH in the same populations. OAF stimulated adenyl cyclase in both types of cell populations, but did not produce significant changes in cAMP phosphodiesterase activity. OAF differed from PTH in that its effects on cAMP accumulation decreased sharply at supramaximal doses in both bone and isolated cells, especially in the matrix-derived populations. Bone resorption did not decrease as markedly as did cAMP accumulation at high doses of OAF, suggesting that cAMP accumulation and resorption could be dissociated under some conditions. These results indicate that OAF has effects on cAMP production in the same cell populations as PTH, and suggest that OAF could modify not only resorption but also formation of bone in vivo. OAF may exert its effects on bone by means of cAMP-dependent mechanisms, but more data will be necessary to establish this unequivocally. The observed differences between OAF and PTH may be of relevance in the mechanism and treatment of pathologic bone destruction in vivo.
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  • 50
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    Calcified tissue international 27 (1979), S. 199-204 
    ISSN: 1432-0827
    Keywords: Bone ; Parathyroid hormone ; Calcitonin ; Osteoporosis ; Paraplegia ; Hydroxyproline
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary In paraplegia, osteoporosis below the neurological lesion occurs early after the spinal cord affection. The serum levels of parathyroid hormone (PTH) and calcitonin (CT), using a radioimmunoassay for the measurement of immunoreactivity, were studied in 12 paraplegic patients for 9 months following onset. Serum Ca and P levels, urinary hydroxyproline excretion, and the kinetic metabolic clearance of45Ca have also been measured. P and immunoreactive (i) CT levels were found the highest at the beginning of the observation and progressively decreased with time. Ca and iPTH serum levels varied inversely with time, the highest level of Ca and the lowest level of iPTH being recorded at the third month following the paraplegia. Mean values of Ca, iPTH, and iCT were in the normal range throughout the study. P levels were increased during the first 3 months. Hydroxyprolinuria was also high and45Ca kinetics showed increased values of Vt, Vo+, and Vu. These parameters indicate a high degree of bone turnover. The results were consistent with the assumption that PTH is not responsible for the increased resorption of bone in paraplegia. Likewise, a deficiency of CT does not seem to be responsible for this bone resorption. These endocrine modifications could be secondary to an increase in the calcium flux from bone to blood and resulting from bone destruction as attested by the increase of urinary calcium and urinary hydroxyproline excretion.
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  • 51
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    Calcified tissue international 27 (1979), S. 233-237 
    ISSN: 1432-0827
    Keywords: Bone ; Vitamin D ; Acidosis ; Phosphate
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    Topics: Biology , Medicine , Physics
    Notes: Summary Vitamin D and phosphate deficiency were produced in rats in order (a) to evaluate the degree of bone mineral and matrix maturation using a bromoform/toluene density gradient technique; and (b) to compare the aforementioned bone maturational changes due to vitamin D and phosphate deprivation to those produced with superimposed severe acidosis. Rats were fed a diet deficient in vitamin D and phosphorus (0.2%) from 3 weeks through 7 weeks of age. To examine the additional contribution of dietary calcium, we gave one-half of the animals either a low (0.06%) or high (1.3%) calcium diet. Following the 4 weeks of vitamin D deficiency, one-half of each group was given 1.8% NH4Cl in the drinking water for 4 succeeding days to induce an acute, severe acidosis. The degree of bone maturation was quantitated via bromoformtoulene density gradient fractionation; total mineral and hydroxyproline (collagen) levels were quantitated as well. The vitamin D-deficient rats deprived of adequate dietary phosphate responded by conserving phosphorus, and as a consequence total bone phosphorus levels were maintained within that level for control rats. This conservation was independent of calcium intake but was extremely sensitive to acute acid loading, where a significant reduction in total bone phosphorus was noted. The bone maturational profile obtained from the vitamin D-phosphate deficient rats, however, revealed a significant accumulation of less mature or dense bone collagen and mineral with a corresponding decrease in the most mature or dense moieties. In contrast to the reduction of the total bone phosphorus content by acute acidosis, the skeletal collagen-mineral maturational profile was not significantly affected by the short-term systemic acidosis. The observed retardations in the bone collagen and mineral maturation of the vitamin D-deficient, phosphate-deprived state provide an additional observation which may well relate to the progressive osteopenia documented in states of chronic, mild acidosis.
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  • 52
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    Calcified tissue international 27 (1979), S. 275-279 
    ISSN: 1432-0827
    Keywords: Proline ; Cartilage ; Bone
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    Topics: Biology , Medicine , Physics
    Notes: Summary Proline biosynthetic and degradative enzymes are unevenly distributed in differentiated mammalian tissues. Activities of the synthetic enzymes are relatively high in collagenous tissues, whereas activities of the degradative enzymes are high in noncollagenous tissues. In order to further characterize tissue-specific proline biosynthesis and degradation, we have determined proline enzyme activities during cartilage and bone formation induced by demineralized bone matrix. We can thus follow temporal changes in enzyme activity in a single tissue as different cell types develop. Ornithine aminotransferase and pyrroline-5-carboxylate reductase have peaks of activity which correlate with maximal type II collagen synthesis by chondrocytes. Both enzymes also are active during bone formation. In contrast, proline oxidase and pyrroline-5-carboxylate dehydrogenase are present at low levels and do not change as new cell types appear. Arginase activity peaks during the first 3 days and then rapidly decreases by the time cartilage and bone formation begin. These observations further substantiate the importance of proline biosynthesis in collagenous tissues. The close correlation between ornithine aminotransferase activity and type II collagen synthesis suggests that the pathway from ornithine to proline may be especially important during formation of type II collagen.
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  • 53
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    Calcified tissue international 25 (1978), S. 227-232 
    ISSN: 1432-0827
    Keywords: Parathyroid hormone ; Bone ; Calcium ; Chick embryo ; Organ culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The purpose of this study was to measure in an in vitro system the movement of Ca and phosphate (Pi) out of bone when treated with parathyroid hormone (PTH). Tibiae from 13-day chick embryos were incubated for up to 8 h in a defined medium containing 1.8 mM Ca. Medium samples were collected every 2 h and were analyzed for Ca, Pi and lactate. Net effluxes from the bones were calculated. When bones were incubated with PTH in the medium (1 U/ml), net Ca efflux was increased 44, 60 and 100% at 4, 6 and 8 h, respectively. At no time was net Pi efflux affected by the hormone. The well known PTH-stimulated lactate production was not seen until 8 h. Lower doses of PTH (0.1 and 0.3 U/ml) were also effective. Comparing PTH (1 U/ml) responsiveness at higher (2.2 mM) and lower (0.9 mM) medium Ca concentrations, showed that with 2.2 mM Ca no increased Ca efflux was seen, while with 0.9 mM Ca significant elevation in medium Ca occurred 2 h sooner than in the experiments using 1.8 mM Ca. In another experiment, varying the medium Pi level from 1 to 2 mM had no effect on the Ca response to PTH. In neither experiment was Pi release affected by PTH. The results of this study have led to the following conclusions: (1) PTH acts on bone to cause an early dose related increase in net Ca efflux; (2) the effect is specific for Ca, since it is not accompanied by an increased Pi efflux, and may be saturated by raising the medium Ca level; and (3) PTH-stimulated Ca efflux in this system is not correlated with, and is probably not a result of increased lactate production.
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  • 54
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    Calcified tissue international 25 (1978), S. 233-240 
    ISSN: 1432-0827
    Keywords: Bone ; Bone resorption ; Albumin
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    Topics: Biology , Medicine , Physics
    Notes: Summary A fraction (brA), which causes resorption of fetal rat bones in vitro, has been concentrated from bovine serum albumin by anion exchange column chromatography on DEAE Sephadex. This active fraction has also been prepared using DEAE Sephadex A-50 by a batch method with a 0.09M NaCl, 0.1M TRIS buffer, pH 8.35. BrA was 10–30 times more potent than the original albumin. The retained material, which constitutes the bulk of the protein and has less activity than the original albumin, elutes with 0.45M NaCl. Similar treatment of serumα,β or γ globulins does not yield brA. Further enhancement of the bone resorbing activity of brA can be obtained with (NH4)2SO4 fractionation or extraction with CH3OH∶CHCl3. Heating at 55° C for 2 h or at 100° C for 10 min does not affect the activity; overnight incubation with protease destroys the bone resorbing effect. The bone resorbing activity is not removed by dialysis and does not correlate with the protease activity of the fraction. The action of brA is inhibited by 3 mM PO4, 1 μg/ml calcitonin or glucagon, 10−7 M dexamethasone or 0.02 μg/ml actinomycin D. The bone resorbing activity of brA is partially inhibited by 10−7–10−5 M indomethacin. PTH did not elicit bone resorption when added to cultures incubated in chemically defined medium supplemented with 0.1 mg/ml brA. However, brA did not inhibit PTH-induced resorption.
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  • 55
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    Calcified tissue international 32 (1980), S. 123-133 
    ISSN: 1432-0827
    Keywords: Bone ; Cell kinetics ; Osteoprogenitor cell ; Migration ; Differentiation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The purpose of this work was to provide further knowledge about bone cell kinetics in the metaphysis of the growing long bone. Seventy rats were sacrificed from 1 to 120 h after injection of tritiated thymidine. Autoradiographs of 3 µm thick sections of the proximal tibial metaphysis were studied in a manner which allowed evaluation of labeled cell nuclei as a function of increasing age of metaphyseal tissue. A cell cycle duration study for osteoprogenitor cells was done. Labeled osteoprogenitor cells and osteoblasts first appeared at 1 h post-injection. The great majority of all labeled osteoprogenitor cells and osteoblasts was found within 1 mm of the growth cartilage-metaphyseal junction (GCMJ) at all times, apparently migrating with the moving GCMJ. In contrast, labeled osteoclast nuclei first appeared at 24 h post-injection within 0.3 mm of the GCMJ and remained always with the area of bone surface with which they were first associated, even as the GCMJ migrated away. By 5 days post-injection, the source of new labeled osteoclast nuclei in the metaphysis near the GCMJ was depleted, whereas that for the osteoblasts remained. The existence of two kinetically different, as well as ultrastructurally different, members of the metaphyseal osteoprogenitor cells population is postulated. A cell cycle time of 39±18 h was found for the osteoprogenitor cell population, but has limited meaning. A schema for metaphyseal bone cell movements during longitudinal bone growth is presented.
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  • 56
    ISSN: 1432-0827
    Keywords: APD ; Bisphosphonate ; Bone ; Resorption ; Formation
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    Topics: Biology , Medicine , Physics
    Notes: Summary Dose-related effects of APD on bone metabolism and Ca homeostasis were studied in rats. The experimental approach consisted of longitudinal and cross-sectional observations, aiming at a kinetic interpretation. Bone and cartilage resorption was inhibited within 2–8 days at doses between 0.16 and 16 µmol/kg body weight/day. This was followed by changes in bone apposition that needed at least 23 days for a maximal effect. The time lag created a transient dissociation between resorption and apposition resulting in excess Ca and P retention, adding to increased metaphyseal bone mass. At high doses of APD (≥40 µmol/kg/day) the mineral content of new matrix decreased, associated with impairment of longitudinal growth of long bones. It is concluded that the lower doses of APD inhibited resorption of bone and cartilage, possibly by physicochemical stabilization of bone mineral, whereas the effect on bone apposition was due to a cellular homeostatic mechanism. Inhibition of growth and of matrix calcification, requiring much higher doses, may be due to a direct, toxic effect on bone cells. The modes of action of APD are discussed in relation to EHDP and Cl2MDP.
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  • 57
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    Calcified tissue international 32 (1980), S. 195-199 
    ISSN: 1432-0827
    Keywords: Bone ; Diabetes ; Insulin
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    Topics: Biology , Medicine , Physics
    Notes: Summary A simple instrument is described which measures the breaking strength of rat bones. The apparatus yields reproducible results and is suitable for use in measuring the strength of bones from both large and small animals. Diabetic rat femurs were more fragile and required less force to break in contrast to those from diabetic rats treated with insulin or normal rats. Daily insulin treatment significantly improved the bone cortical thickness and enhanced their capacity to withstand pressure, although these did not reach the level of the normal controls. The amount of force required to break the bone appears to be related to its cortical thickness and mass.
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    Calcified tissue international 32 (1980), S. 189-194 
    ISSN: 1432-0827
    Keywords: Bone ; Fluoride ; Strength ; Porosity ; Mineralization
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    Topics: Biology , Medicine , Physics
    Notes: Summary Radiographically normal vertebral bone cylinders from 80 male subjects were tested mechanically by static compression and analyzed for porosity, fluoride and ash content. As a group, they had low fluoride content, suggesting little prior intake, consonent with this geographic area. Nevertheless, increasing levels of fluoride were associated with bulkier bone, less porosity, and less mineral per unit of bone, which in direction though not degree suggested changes similar to those of osteomalacia and opposite from those of osteoporosis without apparent threshold. The higher fluoride hard tissue was weaker in static tests than that with less fluoride, but the increased bulk apparently offset this, resulting in bones of unchanged static strength. Hence, water fluoridation should not alter static bone strength. There has, however, been a recent report suggesting that increased mineralization of bone renders it more brittle and thus more likely to fracture on impact. Therefore, the possibility that fluoridation may increase impact resistance by lessening mineralization can be entertained.
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  • 59
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    Calcified tissue international 32 (1980), S. 229-236 
    ISSN: 1432-0827
    Keywords: Bone ; Ion influxes ; Calcium ; Phosphate ; Exchange
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    Topics: Biology , Medicine , Physics
    Notes: Summary Quantitative measurements were made of the ion fluxes of calcium and phosphate into and from calvaria (mouse or rat) when clamped in specially designed micro-Ussing chambers. The effects of varying concentrations of calcium were examined on the influx and efflux of calcium and of its counterion, phosphate. A comparable series of experiments was performed with varying phosphate concentrations. Both ions, as their concentrations increased, depressed their own influx, increased their own efflux, and significantly increased the equilibrium concentration, E/K, supported by the calvaria. Similarly, both ions, as their concentrations increased, affected the influx or efflux of their counterion only slightly but did depress the counterion's equilibrium level, E/K, significantly. In spite of these changes it was shown that calvaria effectively buffered the medium at physiological concentrations of calcium and phosphate. The buffering capacity, however, was small, and the balance, E/K, was modified by small uptake or loss of either ion. The small size of the interacting mineral pool was confirmed by direct measurement of the rapidly exchanging fractions of both calcium or phosphate. They were only ∼1% of the total ions present. The significance of these findings is discussed.
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  • 60
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    Keywords: Bone ; Calcium metabolism ; Whole-body counting
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    Topics: Biology , Medicine , Physics
    Notes: Summary A whole-body counter was used to measure the retention and distribution of radioactivity along the longitudinal axis of the body during 10 days after the intravenous injection of 50 µCi of47Ca. These data and the simultaneous measurements of the serum specific activity allowed us to calculate, by the Bauer-Carlsson-Lindquist (BCL) formulas generalized by Marshall, the calcium accretion rate and exchangeable pool in 7 areas: skull, thorax, pelvis, thighs, knees, legs, and ankles and feet. For the whole body, the accretion rate was 336±115 mg/24 h, and the exchangeable pool 5668±1221 mg, in 26 subjects without bone disease. Both parameters were significantly correlated with body height; the exchangeable pool was significantly higher in men than in women. The accretion rate and exchangeable pool expressed on a basis of bone content varied widely from one area to another, reflecting the ratio of spongy to compact bone: the thorax, pelvis, and arms, which contain 49% of the skeletal mass, accounted for 57% of the total accretion rate and 53% of the exchangeable pool; the skull, 18% of the skeletal mass, accounted for 14% of the accretion and 11% of the pool.
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  • 61
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    Keywords: Dentin ; Bone ; Apposition rate ; Parathyroid hormone
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    Topics: Biology , Medicine , Physics
    Notes: Summary The dose-dependency of the effects of parathyroid hormone (PTH) on bone and dentin apposition after both intermittent and continuous administration of the hormone was investigated. The purpose was to compare the sensitivity of these two mineralizing tissues to parathyroid hormone and to provide additional information regarding the direct effect of PTH on mineralized tissue formation. Adult rats were thyroparathyroidectomized and 5 groups of 4 or 5 rats each were given daily subcutaneous injections with different doses of bovine parathyroid hormone. Five more groups of 4 rats each were administered equivalent dosages by means of a continuous infusion pump implanted subcutaneously. An additional group of 4 rats served as controls. All animals were labeled with tetracycline injected on days 7, 9, and 11. The animals were killed on day 12 and blood samples were collected for serum calcium determination. The lower metaphysis of the femur and the left and right mandibles were dissected out, and undecalcified sections of plastic-embedded tissues were prepared. The distances between the three tetracycline bands were measured to determine the amount of bone or dentin formation. Results indicated that both dentin and bone apposition increased with higher dosages of hormone. No overall effect of the method of administration was evident. For both methods, bone apposition showed a more pronounced increase over the control levels than dentin apposition. This suggests that, although both osteoblasts and odontoblasts appear to respond directly to PTH, differences do exist in the magnitude and dose-dependency of the response. No causal relationship was found between increases in serum calcium levels and either bone or dentin apposition at the lower dose levels.
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  • 62
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    Calcified tissue international 35 (1983), S. 728-731 
    ISSN: 1432-0827
    Keywords: Bone ; Bioelectricity
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    Topics: Biology , Medicine , Physics
    Notes: Summary The origin of the bioelectric potentials in unstressed living bones is still an open question. Blood-flow in vessels, stationary potentials on peripheral nerves, muscle injury potentials, and viability of bone cells are claimed to be possible sites of origin of the electric potentials recorded on bone surface. The present data show that the topographic quantitative distribution of tetracycline labeling at sequential levels of rabbit tibia and the distribution pattern of the bioelectric potentials in this bone are significantly superimposable. This coincidence of bone formation rates and bioelectric potentials seems to support the view that the latter are in some way linked with the laying down of new bone.
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  • 63
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    Keywords: Bone ; Osteosarcoma ; Matrix vesicles ; Alkaline phosphatase ; Aminopeptidase ; Naphthylamidase
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    Topics: Biology , Medicine , Physics
    Notes: Summary Extracellular matrix vesicles from bovine fetal alveolar bone and from a dog osteosarcoma were isolated by differential centrifugation and then fractionated on a discontinuous sucrose density gradient. The fractions were examined by electron microscopy and were analyzed for protein, alkaline phosphatase, aminotripeptidase, and four different β-naphthylamidase activities. The low-density peak of enzyme activities was shown by electron microscopy to be much more homogeneous than the crude matrix vesicle fraction. Two major peaks of protein and enzyme activities were present, one in the high and one in the low density layers. There was good correlation between the activities of alkaline phosphatase and the various peptidases in the fractions from the sucrose density gradient. These results indicate a coexistence of peptidase and alkaline phosphatase in matrix vesicles. On the other hand, there was generally no correlation between the peptidase and alkaline phosphatase activities in vesicular specimens from bovine liver obtained in the same way. Most of the peptidase activity and about half of the alkaline phosphatase activity were solubilized from bone matrix vesicles by detergents. The extracted alkaline phosphatase and alanyl β-naphthylamidase activities were separated from each other on a DEAE-cellulose column.
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    Calcified tissue international 36 (1984), S. S19 
    ISSN: 1432-0827
    Keywords: Bone ; Remodeling ; Microdamage
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    Topics: Biology , Medicine , Physics
    Notes: Summary A conceptual framework is presented for understanding and investigating structural adaptation of cortical bone. The magnitudes, orientations, and sense (tension or compression) of the physiologically incurred cyclic principal strains vary markedly throughout the skeleton. It is probable, therefore, that the strain/remodeling response of bone is site specific. Furthermore, there is some indication that immature bone is more responsive to alterations of cyclic strains than mature bone. Animal experimental studies and complementary stress and strain analyses suggest that the structural adaptation due to changes in cyclic strain fields may be a very nonlinear response. Bone loss in mature animals due to immobilization is sensitive to even small changes in the cyclic bone strains. Under normal conditions, however, there appears to be a broad range of physical activity in which bone is relatively unresponsive to changes in loading history. With severe repeated loading, bone hypertrophy can be pronounced. These observations open the possibility that bone atrophy and hypertrophy are controlled by different mechanisms. Therefore, two (or more) complementary control systems may be involved in the regulation of bone mass by bone cyclic strain histories. It is probable that bone mechanical microdamage is one control stimulus for affecting an increase in bone mass.
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    Calcified tissue international 34 (1982), S. 519-522 
    ISSN: 1432-0827
    Keywords: Turner's syndrome ; Ovarian dysgenesis ; Osteoporosis ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The bone mineral status of 17 girls with Turner's syndrome was evaluated by single photon absorptiometry. Bone mineral content (BMC) was 25.4% below that predicted by normalization for age, sex, height, weight, and bone width. Only 25% of this demineralization could be attributed to delayed skeletal maturation. Bones of girls who received estrogen replacement therapy were less demineralized than those of the others. The bone mineral deficit became less pronounced with advancing age. It could not be determined if the apparent effect of estrogens was related to age or if the apparent improvement with age was really due to an effect of estrogen treatment. For 8 subjects followed longitudinally there was no significant change in the BMC deficit.
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  • 66
    ISSN: 1432-0827
    Keywords: Maize ; Vitamin D deficiency ; Bone ; Rickets ; Osteomalacia
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    Topics: Biology , Medicine , Physics
    Notes: Summary Three groups of young baboons were fed for 16 months on one of three diets. The first group was given a well-tried semisynthetic formula, the second group the same diet save that vitamin D had been omitted, and the third group was given the vitamin D-free diet in which maize replaced the dextrin normally used. Although both groups fed the vitamin D-free diets developed rickets and osteomalacia, the group receiving maize did so far more rapidly and to a much greater degree of severity, as evidenced by clinical, radiological, biochemical, and histological signs. The mechanism by which maize acts remains unclear, but this report serves to emphasize the extremely detrimental effects that might be expected in populations who are deficient in vitamin D and who have predominantly cereal diets.
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    Calcified tissue international 33 (1981), S. 57-69 
    ISSN: 1432-0827
    Keywords: Thallium ; Chondrodystrophy ; Cartilage ; Bone ; Matrix
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Using the transmission electron microscope, we sought to describe the morphology of thallium sulfate-induced chondrodystrophy in chick embryos. There was cell death and degeneration in all zones of growth cartilage, but the cells and matrix of the hypertrophic zone were the most severely affected. Ultrastructural changes of the hypertrophic chondrocytes consisted of alteration of the cytoplasmic contents and of the intercellular matrix; the cell membrane was smooth and without cytoplasmic extensions. The cytoplasm was filled with dilated rough endoplasmic reticulum, vacuoles of varying sizes and contents, and lipidlike bodies with electron-dense granules; mineral crystals, collagen, and degenerating mitochondria were present. The matrix showed only spotty calcification and a reduced number of dense bodies, vesicles, and granules. The cells appeared to have failed to exteriorize cell products across the plasmalemma. Failure to exteriorize cell products and to form cytoplasmic processes reduced the number of potential nucleation sites for calcification. The ultrastructure of osteocytes was much less affected.
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    Calcified tissue international 33 (1981), S. 89-99 
    ISSN: 1432-0827
    Keywords: Soluble proteoglycans ; Resistant proteoglycans ; Collagen ; Bone ; Cartilage
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    Topics: Biology , Medicine , Physics
    Notes: Summary Soluble proteoglycans (SPG) were extracted from bovine (BCC) and human (HCC) costal cartilages by the dissociative method using 4 M guanidinium chloride (GuHCl). Proteoglycans which are resistant to extraction (RPG) were obtained following collagenase digestion or hydroxylamine treatment of the cartilage residues. Similarly, SPG were extracted from bovine metaphyseal and cortical bone using EDTA. The RPG were extracted from the bones using hydroxylamine. Density gradient fractionation under dissociative conditions of cartilage SPG and RPG followed by chromatography on Sepharose 2B revealed that A1D1 RPG are smaller than the SPG. SPG reacted with either collagenase or hydroxylamine are also smaller than the parent SPG. A1D1 fractions obtained from BCC-SPG and RPG or from mixtures of SPG and acid-soluble collagen are free of hydroxyproline. Hydroxyproline is not completely separated from HCC-RPG. Density gradient fractionation of bone proteoglycans and Sepharose chromatography of the A1 and A1D1 fractions showed that those obtained from metaphysis are larger than those from cortical bone. This was attributed to the presence of calcified cartilage in metaphyseal bone. The A1D1 fractions of the metaphyseal proteoglycans seemed to undergo self-association since this fraction is larger than the A1 fraction from which it is derived. Cortical bone proteoglycans do not behave similarly. Density gradient purification under dissociative conditions failed to separate hydroxyproline from the proteoglycans obtained from bone. It is hypothesized that in bone proteoglycans and collagen might be linked.
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  • 69
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    Calcified tissue international 33 (1981), S. 567-574 
    ISSN: 1432-0827
    Keywords: Bone ; Femur ; Age ; Mineral ; Apatite
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary It has previously been observed that after the third decade of life the tensile strength of bone decreases faster than its density [1], indicating that some microstructural change occurs in bone with advancing age. In order to try to elucidate this observation, the orientation and particle size of the mineral phase of bone have been studied with respect to age. Samples were taken from femora varying in age from 13 to 97 years. The results show that the degree of preferred orientation does not vary with age after the first decade of life. However, the percentage of small particles increased up to the fourth decade of life and thereafter decreased, a pattern of change similar to that observed for bone strength. It would seem to indicate that an increase in mineral particle size is a factor that contributes to the loss of bone strength and that an increased percentage of large particles in the later years of life could, along with osteoporosis, help to explain the increased fragility in the bones of the elderly.
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  • 70
    ISSN: 1432-0827
    Keywords: Bone ; Calcium ; Strontium ; Apposition ; Tracer analysis
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    Topics: Biology , Medicine , Physics
    Notes: Abstract 1. Evidence has accumulated that the rate of acceretion (A) of calcium to bone is the sum of two fluxes; apposition involving the laying down of new bone and augmentation which is the result of slow exchange of non-surface bone calcium with plasma calcium pools as the result of solid state diffusion. 2. A method has been devised for separating A into its two components. It requires the use of45Ca or, for clinical studies,85Sr as a calcium tracer. Studies which are initiated with a combined accretion rate-calcium balance study, are concluded with an estimate of the exponent of the power function which has been found to describe the whole body retention of tracer from the second month onward. 3. The impulse response function of the skeleton for the tracer is then calculated, making the assumption that in any uniform volume of bone, osteoclastic resorption is a first order process. Making in addition certain simplifying assumptions, which are shown to have a modest influence on the final results, a mean rate of bone resorption can be calculated using a development of the well known Stewart-Hamilton formula. The apposition rate is calculated as the sum of the resorption rate and the calcium balance. Augmentation and diminution, defined as equal and opposite exchange processes, are given by the difference between A and the apposition rate. 4. The results of our first thirteen studies in normal subjects and patients with metabolic bone disease are presented, together with analyses of some data from the literature. It is concluded that the development of an atraumatic method for measuring rates of bone formation and resorption in the whole body would be an important advance in the study of metabolic bone disease, and this work is presented so that critical comparisons may be initiated between this tracer method and independent histological methods for measuring these parameters.
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  • 71
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    Calcified tissue international 25 (1978), S. 249-254 
    ISSN: 1432-0827
    Keywords: Bone ; Oestrogens ; Oestrogen receptors ; Binding proteins
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    Topics: Biology , Medicine , Physics
    Notes: Summary Attempts were made to demonstrate the presence of specific oestrogen binding proteins (“receptors”) in bone cells. High speed cytosol preparations of bone were incubated with several concentrations of radioactive oestradiol alone and with radioactive oestradiol in the presence of a specific antioestrogen, Nafoxidine. Separation of bound and free oestradiol was carried out by dextran coated charcoal treatment and by sucrose gradient ultracentrifugation. Several types of bones likely to be oestrogen-sensitive were investigated: human femoral heads, human phalanx, rat and rabbit calvaria, humeri and femora of female rats. In all experiments we were unable to demonstrate the presence of specific oestrogen receptors in bone cell cytosol indicating that the direct effect of oestrogens on bone, if present, is not mediated by specific oestrogen receptors.
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  • 72
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    Keywords: Electron spin resonance ; Crystallinity ; Bone ; Osteons
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    Notes: Summary Several groups containing 10–15 isolated osteons differing in their degree of maturity were analysed. Samples were isolated from undecalcified human and bovine bone sections. The crystallinity coefficient, defined as the ratio of the number of radiation-induced paramagnetic defects in the crystalline lattice of hydroxyapatite to the total ash content, was calculated. The results were compared with measurements performed on fragments of total cortical bone, primary periosteal bone, and inner circumferential lamellar bone. The results show a higher crystallinity of fully calcified osteons as compared with that found at the initial stage of calcification. No differences in the ash content were observed between human osteons, from different stages of calcification evaluated morphologically and by X-ray absorption. These differences were evident when bovine osteons differing in their stage of calcification were compared. Human fully calcified osteons contain 60% ash and their crystallinity coefficient is 52.1. Human osteons at the initial stage of calcification contain 57% ash and their crystallinity coefficient is 40.6. The same parameters for fully calcified bovine osteons and for bovine osteons at the initial stage of calcification are 59% ash, 62.6 crystallinity coefficient and 46% ash, 43.0 crystallinity coefficient, respectively.
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  • 73
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    Calcified tissue international 24 (1977), S. 65-72 
    ISSN: 1432-0827
    Keywords: Bone ; Strength ; Tension ; Microstructure ; Osteon
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Standardized human and beef femoral compact bone specimens were tested in tensile impact and the dynamic mechanical properties were determined. The microstructure of 45 beef and 47 human bone specimens were examined histologically to determine if there is a structural basis to account for strength differences in the bone samples. Strong negative correlations were obtained between the maximum stress and the percentage area of secondary osteons in each specimen. For human bone samples, the energy absorption capacity and the modulus of elasticity were also found to have strong negative correlations with the percentage area of secondary osteons present in each specimen. Linear regression equations were obtained describing the impact strength properties in terms of the percentage areas of secondary osteons and cavities in the samples. Fracture surfaces of the tested bone specimens were examined in a scanning electron microscope. Most surfaces exhibited a fairly rough texture indicating a quasi-cleavage type of failure. Fractographic analysis of bone fracture surface was helpful in understanding the micromechanics of bone fracture.
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  • 74
    ISSN: 1432-0827
    Keywords: Bone ; Density ; Composition ; Renal osteodystrophy ; Dogs
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    Notes: Summary Bone density and composition were studied in trabecular and cortical bone from control 2 and 4 year old beagles and those which had various degrees of renal failure as a result of perinatal irradiation. Changes in the two types of bone were qualitatively similar but consistently greater in trabecular bone. In a group identified as markedly uremic, the decrease (P〈0.025) from control levels in specific gravity was about 4 times greater in trabecular than in cortical samples. The decrease (P〈0.025) in grams of ash/ml was 9% in trabecular bone and 2% in cortical bone. These changes were associated with an increase (P〈0.001) in water content and, on a percent by volume basis, approximately equal decreases in ash (P〈0.025) and combustible matter (P〈0.025). In a mildly uremic group there were similar trends in mean values but the only significant difference was an increase (P〈0.001) in trabecular bone water.
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  • 75
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    Calcified tissue international 24 (1977), S. 201-208 
    ISSN: 1432-0827
    Keywords: Calcitonin ; Plasma calcium ; Radiocalcium ; Bone
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    Topics: Biology , Medicine , Physics
    Notes: Summary The effect of intravenous salmon calcitonin (SCT) infusion on the net rate of loss from plasma of recently injected45Ca has been studied in conscious fasted rats, some of which had been previously thyroparathyroidectomized (TPTX). The45Ca was injected 10 h prior to or 1 or 2 h after the initiation of the infusion. Salmon calcitonin (0.05–0.2 MRC mU/g body weight/h) infusion produced the expected hypocalcemia and hypophosphatemia. The drop in plasma calcium levels produced by the hormone was lowered further by the addition of phosphate to the infusate. Plasma45Ca changes, during SCT infusion, followed a biphasic pattern when the radionuclide injection preceded SCT infusion by 10 h. During the first part of the infusion, the rate of loss of45Ca from plasma was greater in SCT-treated than in control rats. Later, this pattern reversed, and the rate of loss of45Ca from plasma became slower in SCT-treated animals. If45Ca was injected after SCT infusion was initiated only the second phase of the biphasic effect was observed; i.e., disappearance of45Ca from plasma was slower in SCT-infused than in control rats. These data are considered to support the hypothesis that rapid and opposing calcium fluxes exist between bone fluid and plasma, and that calcitonin acts by decreasing the calcium efflux from bone fluid. The calcium moved in this efflux system includes some that has only recently entered bone fluid from plasma. This is the explanation given for the initial decrease in plasma45Ca concentrations following SCT infusion. The reverse effect in the second phase can be explained as the result of a secondary reduction in calcium influx to bone fluid from plasma.
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  • 76
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    Calcified tissue international 23 (1977), S. 113-114 
    ISSN: 1432-0827
    Keywords: Bone ; Absorption Spectra ; Collagen ; Hydroxyapatite
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    Topics: Biology , Medicine , Physics
    Notes: Summary Absorption spectra of human bone and its two major constituents (collagen and apatite) were obtained in the wavelength region extending from 2000 to 12,000 Å. In the last two cases a, minimum transmission (maximum absorption) was uniformly observed in the ultraviolet region. The two samples after exposure to ultraviolet radiations show a shift in the peak positions. Absorption peaks in the total bone spectra are not reproducible in its two constituents. After exposure to UV radiations the position of maximum absorption is displaced. A possible interpretation of the observed results is presented.
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  • 77
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    Keywords: Parathyroidectomy ; Diphosphonates ; Bone ; Collagen ; Hydroxyproline
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    Notes: Summary Disodium ethane-1-hydroxy-1,1-diphosphonate (EHDP) may impair mineralization and reduce turnover of bone. However, its administration has been shown to lead to an accumulation of osteoid in both man and the rat. Using rats, the effects of surgical parathyroidectomy and administration of EHDP (30 mg EHDP/kg body weight by subcutaneous injection daily for 5 days) upon bone composition, uptake of3H-proline into hydroxyproline in bone and the subsequent urinary excretion of labelled and unlabelled hydroxyproline have been compared. Both parathyroidectomized (PTX) rats and EHDP-treated animals accumulated more hydroxyproline in bone and excreted less total urinary hydroxyproline than sham-PTX control animals, the changes induced by EHDP being of greater magnitude than those induced by surgical parathyroidectomy. Skeletal buildup of hydroxyproline was associated with decreased catabolism of mature collagen rather than enhanced synthesis of hydroxyproline in bone. EHDP administration, although decreasing urinary excretion of newly synthesised hydroxyproline, did not alter the skeletal synthesis of hydroxyproline as measured by uptake of3H-proline. In all parameters measured, responses to EHDP were similar in PTX and sham-PTX rats. It is concluded that depression of parathyroid hormone mediated bone turnover may have contributed to, but did not fully account for, the lowering of bone turnover and changes in bone composition induced by EHDP in sham-PTX rats in the present experiment.
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  • 78
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    Calcified tissue international 26 (1978), S. 85-89 
    ISSN: 1432-0827
    Keywords: Bone ; Histology ; Microradiography ; Densitometry ; Computer
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    Topics: Biology , Medicine , Physics
    Notes: Summary A comparatively simple quantitative method for assessing bone morphology has been evolved. Microradiographs of thin sections of mandible have been scanned with a Joyce-Loebl double beam recording microdensitometer with a scanning autodensidater attachment, using a white beam. For each image the optical densities for all the pixels (picture elements) were divided into 10 groups. The limits of the division were fixed by the maximum and minimum densities occurring within the image. A computer generated map was produced which indicated the spatial distribution of the pixels within each group to which an arbitrarily chosen shading was attached. The number of pixels within each group is also shown on the map. The computer map was compared with the photomicrograph and, where necessary, the original section. The fractional area of hard tissue was then readily determined using the numerical values of each group of pixels.
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    Calcified tissue international 26 (1978), S. 139-142 
    ISSN: 1432-0827
    Keywords: Chemisorption ; Surface area ; Hydroxyapatite ; Bone ; Enamel
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    Topics: Biology , Medicine , Physics
    Notes: Summary The surface areas of three different samples of hydroxyapatite, samples of deproteinized bone, and samples of whole and deproteinized enamel were determined by adsorption of an adduct (the diglycidyl ether of bisphenol A with N-phenylglycine) from methylene chloride solution. In all cases, the surface areas of these samples agree well with those obtained by the BET (N2) method.
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    Calcified tissue international 27 (1979), S. 109-115 
    ISSN: 1432-0827
    Keywords: Bone ; Bending rigidity ; Disuse atrophy
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    Topics: Biology , Medicine , Physics
    Notes: Summary The in vivo bending rigidity and bone mineral content of monkey ulnae and tibiae were measured. Bending rigidity in the anteroposterior plane was measured by an impedance probe technique. Forced vibrations of the bones were induced with an electromechanical shaker, and force and velocity at the driving point were determined. The responses over the range of 100–250 Hz were utilized to compute the bending rigidity. Bone mineral content in the cross section was determined by a photon absorption technique. Seventeen male monkeys (Macaca nemestrina) weighing 6–14 kg were evaluated. Repeatability of the rigidity measures was 4%. Bone mineral content was measured with a precision of 3.5%. Bending rigidity was correlated with the mineral content of the cross section,r=0.899. Two monkeys were evaluated during prolonged hypodynamic restraint. Restraint produced regional losses of bone most obviously in the proximal tibia. Local bone mineral content declines 17 to 24% and the average bending rigidity declines 12 to 22%. Changes in bones leading to a reduction in mineral content and stiffness are discussed.
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  • 81
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    Calcified tissue international 25 (1978), S. 105-110 
    ISSN: 1432-0827
    Keywords: Progesterone ; 17β estradiol ; Testosterone ; Tissue culture ; Bone
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    Topics: Biology , Medicine , Physics
    Notes: Summary Although sex steroids are known to affect skeletal metabolism, their effects on bone collagen synthesis have not been studied. We have examined the direct effects of progesterone, 17β estradiol, testosterone, 5α dihydrotestosterone and dehydroepiandrosterone on bone collagen and noncollagen protein synthesis in cultures of calvaria obtained from 21-day fetal rats. Bones were incubated for 24 to 168 h and3H-proline was added for the last 2 h of culture. Incorporation of the label into collagenase-digestible protein (CDP)1 and noncollagen protein (NCP) was determined using repurified bacterial collagenase. Progesterone caused a dose dependent inhibition of the labeling of CDP at concentrations of 3×10−7 M to 10−5 M after 96 h of culture. A smaller inhibitory effect was observed on NCP. The inhibitory effect was slow in onset and persisted when bones were incubated for 48 h with progesterone and then transferred to control medium for 48 h. Progesterone also inhibited the incorporation of3H-thymidine and3H-uridine into fetal rat calvaria. After 24 h of culture, 17β estradiol and testosterone had no effects on the labeling of CDP and NCP. After 96 h, 17β estradiol had a small and inconsistent stimulatory effect on the labeling of CDP but testosterone had no effect. Neither hormone altered the inhibitory effects of parathyroid hormone, cortisol and progesterone. Dihydrotestosterone and dehydroepiandrosterone had no effects after 24 h and 96 h of culture. 17β estradiol, testosterone and dihydrotestosterone did not affect the incorporation of3H-uridine or3H-thymidine into fetal rat calvaria. Our studies indicate that progesterone is an inhibitor of bone collagen synthesis and estrogens and androgens are not major regulators of bone formation in vitro.
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  • 82
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    Calcified tissue international 27 (1979), S. 13-18 
    ISSN: 1432-0827
    Keywords: Anticonvulsants ; Calcium ; Vitamin D ; Parathyroid hormone ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Parameters of mineral metabolism were examined in 6 patients with moderately severe anticonvulsant drug-induced osteomalacia. Compared to 15 matched controls, the patients exhibited significantly reduced serum calcium, inorganic phosphate, and 25-hydroxyvitamin D concentration, elevated serum alkaline phosphatase and immunoreactive parathyroid hormone (iPTH) concentration, reduced intestinal47Ca absorption, reduced urinary calcium and increased urinary hydroxyproline excretion, and reduced forearm bone mass. Intestinal absorption of vitamin D3 was normal. Following 4 months of treatment with vitamin D3 (4000 units/day), serum 25-OHD concentration was increased to 3 times mean normal values and all parameters except serum iPTH, urinary calcium excretion, and forearm bone mass were returned to levels not significantly different from normal. Serum iPTH concentration was reduced by 39% (P〈0.05); 24-h urinary calcium excretion rose by 98% (P〈0.001), and forearm bone mass increased by 5.6% (P〈0.05). It is concluded that moderate-dose vitamin D3 supplementation is effective in normalizing parameters of mineral metabolism in this disorder, despite evidence of resistance to the biologic effects of vitamin D.
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  • 83
    ISSN: 1432-0827
    Keywords: Calcium ; PTH ; 1,25(OH)2D ; Acid ; Bone
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    Notes: Summary We measured mineral and acid balances, serum iPTH, urinary cAMP/creatinine, and plasma concentrations of 25OHD and 1,25(OH)2D in 7 healthy adults during control conditions and during increased fixed acid production achieved either by the administration of NH4Cl (N=3) or by increased dietary protein intake (N=4). When acid production was increased, the subjects were in positive acid balance and negative Ca balance because of increased urinary Ca excretion. Serum iPTH fell slightly but urinary cAMP and the plasma levels of vitamin D metabolites did not change. We conclude that the accelerated skeletal and urinary losses of Ca that occur when fixed acid production is increased are not contributed to nor compensated for by the parathyroid-vitamin D endocrine systems.
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    Calcified tissue international 30 (1980), S. 15-20 
    ISSN: 1432-0827
    Keywords: Bone ; Osteoclasts ; Macrophage ; Resorption ; Plutonium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Female rats were used to study the kinetics of plutonium transfer from the bone surfaces of the mandibular condyle to osteoclasts and macrophages. This study was made using autoradiographs prepared from plastic sections of the mineralized bones of animals which had been injected with241 Pu citrate. Measurements of the concentration of plutonium in the osteoclasts and macrophages at different times after the injection of plutonium showed that plutonium was concentrated by osteoclasts from bone surfaces and was retained with a half-time of ∼ 70 h. Subsequently, plutonium appeared to be transferred to macrophages. The results showed that plutonium was unlikely to be accumulated by macrophages as a result of their participation in bone resorption.
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  • 85
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    Calcified tissue international 29 (1979), S. 169-171 
    ISSN: 1432-0827
    Keywords: Glycosaminoglycans ; Bone
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    Notes: Summary Compact bone tissue chondroitin sulphate, previously considered to be pure chondroitin-4-sulphate, was isolated from three adult human femoral diaphyses and digested with chondroitinase ABC. Assaying these digests by means of high performance liquid chromatography we could demonstrate that 12 to 14% of the disaccharide residues were 6-sulphated. The 4/6-ratios were also studied in chondroitin sulphate fractions of different molecular size. Slightly increasing amounts of 6-sulphated disaccharides were found with increasing size of the chondroitin sulphates.
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    Calcified tissue international 30 (1980), S. 67-71 
    ISSN: 1432-0827
    Keywords: Alkaline phosphatase ; ATPase ; Bone ; Triton
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    Topics: Biology , Medicine , Physics
    Notes: Summary Extraction with Triton X-100 has proved effective in solubilizing alkaline phosphatase from rat bone particles, whereas ATPase with optimum activity at pH 8 remains attached to the bone particles. The kinetic characteristics of the ATPase activity of the Triton extracts are different from those of the same enzyme attached to bone particles, but the kinetic characteristics of the particle-bound and solubilized alkaline phosphatases are similar. The results suggest that the Triton extracts do not have true ATPase activity and provide a means of separating the ATPase and alkaline phosphatase activities.
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  • 87
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    Calcified tissue international 29 (1979), S. 7-13 
    ISSN: 1432-0827
    Keywords: Bone ; Culture ; Resorption ; Acidosis ; Alkalosis
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    Topics: Biology , Medicine , Physics
    Notes: Summary We have examined the effects of H+, CO2, and HCO3 − concentrations during metabolic and respiratory acidosis and alkalosis on bone resorption in vitro. Rat fetal bones prelabeled with45CaCl2 were cultured at 2%, 5%, and 10% CO2 for up to 120 h, and the release of45Ca was measured in devitalized bones (non-cell-mediated45Ca release) and in live bones (cell-mediated45Ca release) cultured with or without PTH and 1,25(OH)2D3. Non-cell-mediated mineral loss was linearly related to H+ concentration but not to CO2 or HCO3 − concentration. This effect was observed on both labeled and stable calcium. Over a wide pH range (6.9–7.5) H+, CO2, or HCO3 − concentrations did not influence cell-mediated bone resorption in control or in PTH-and 1,25(OH)2D3-stimulated cultures. However, inhibition of cell-mediated bone resorption was observed at higher or lower pH irrespective of CO2 or HCO3 − concentrations. These observations demonstrate that the bone mineral mobilizing effect of acidosis in vitro is mainly due to the effect of changing H+ concentration on devitalized bone. Effects on cell-mediated bone resorption and hormonal response were observed only at extremes of pH. The effects of H+ were independent of changes in CO2 or HCO3 − concentration and could be responsible for the negative calcium balance and increased urinary loss observed in metabolic acidosis in vivo, but do not explain the reported differences in effects on calcium metabolism between respiratory and metabolic acidosis.
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    Calcified tissue international 31 (1980), S. 135-145 
    ISSN: 1432-0827
    Keywords: Bone ; water compartments ; marker ; methods ; pH ; potassium ; lactate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The concept of fluid compartmentalization in bone has emphasized the need for and lack of suitable methodology for the quantitation of water “spaces” in bone, in particular intracellular water, ICF, and extracellular water, BECF. Over a dozen commonly used marker substances were studied intensively. Small columns of hydroxyapatite crystals were employed to reveal physiochemical interactions with the mineral phase, and isotope distributions in live and dead calvaria (rat pup, and adult mice) were employed to evaluate interactions with the combined organic-inorganic matrix of bone itself. The results were most discouraging. For example, for total water space, only water itself (either by direct weight or by3H2O exchange) is a reliable measure in all instances. All markers studied were passively concentrated or excluded to varying degrees. In the end, it was necessary to measure intracellular space “by difference.” An extracellular marker, polyethylene glycol (14C-, mol. wt. 4000), was incubated with viable calvariae and also with comparable calvariae having lysed cells (0.1% Triton X-100). The intracellular space thus determined by difference agreed well with correlative data obtained on mixed cell-isolates from similar specimens. In neonatal calvariae, the intracellular space was 30% of the total water present; that of adult mouse tibia, 18%; and that of adult mouse calvariae, 14.5%. Measurements of dimethyl-2,4-oxazolidinedione (DMO) distribution in live and lysed calvaria revealed an overall pH differential (inside bone vs. medium) of 0.1 unit or less. Using the best available data for ICF, BECF, and the passive concentrating effects of matrix-K+ interactions, there still remained an unexplained excess of K+ in the BECF.
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    Calcified tissue international 31 (1980), S. 215-223 
    ISSN: 1432-0827
    Keywords: Bone ; 1,25(OH)2D3 ; 25OHD3 ; Histology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Rachitic rats, maintained on diets with low or normal P contents, were given daily intraperitoneal doses of 1,25(OH)2D3 or 25OHD3 at levels of 100 or 200 ng. Plasma chemistry was measured and the ash content and histological appearance of the bones investigated. Using labeled material it was shown that the dosing levels of 1,25(OH)2D3 employed ensured a higher than normal plasma concentration of that metabolite over the period between doses. 1,25(OH)2D3 was not as effective as 25OHD3 in raising bone ash or reducing the amount of osteoid. The difference between the effects of the metabolites was evident at both dietary P levels, but more marked at the higher P level. In contrast, the metabolites reduced the width of the epiphyseal plate to an approximately similar degree, and this is possibly the reason why there are discrepancies between previous reports of the effectiveness of 1,25(OH)2D3 compared with 25OHD3 or vitamin D3. Dosing with 1,25(OH)2D3 failed to maintain a constant plasma Pi value over the period between doses in animals fed the low P diet.
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    Calcified tissue international 31 (1980), S. 231-238 
    ISSN: 1432-0827
    Keywords: Magnesium ; Bone ; Calcium ; Parathyroid gland
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    Notes: Summary Calcium homeostasis and bone pathology were studied in weanling rats fed a low (70 ppm) magnesium diet for 2–21 days. The rats developed significant, progressive hypercalcemia after 6 days on the diet. The increase in blood calcium was accompanied by progressive hypoactivity of the parathyroid gland (PTG), as determined by histologic and morphometric analyses. Thus hyperactivity of the PTG could not have been responsible for the hypercalcemia observed. Histologic examination of femora and humeri from magnesium-deficient rats showed progressive subperiosteal hyperplasia, consisting of undifferentiated osteoprogenitor cells and fibrous tissue, after 7 days of deficiency. The presence of unmineralized osteoid tissue in the metaphyses indicated that mineralization was not proceeding normally. The alterations in differentiation of osteoprogenitor cells, together with the failure of mineralization, resulted in significantly lower rates of bone formation (as measured by fluorochrome labeling) in the magnesium-deficient rats. Basophilic cementing lines and inactive osteocytes in the cortices of bones from magnesium-deficient rats indicated that bone resorption was also severely reduced in magnesium deficiency. We postulate that bone magnesium depletion (66% by day 21) has a direct negative effect on osteoblastic and osteocytic activity, and may explain, in part, the decreased responsiveness of bone to parathyroid hormone (PTH) that has been observed in magnesium-deficient animals.
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    Calcified tissue international 31 (1980), S. 261-265 
    ISSN: 1432-0827
    Keywords: Bone ; blood flow ; microspheres ; fluorine ; strontium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Radioactive microspheres have been used for measuring bone blood flow and the extraction ratios of bone-seeking radionuclides, but we have shown that the technique is not completely valid. Bone uptake of18F and85Sr was compared with that of microspheres simultaneously injected into the heart or aorta of rats and rabbits under a variety of conditions. In single-passage experiments with quick killing to avoid recirculation, attention was focused on the tibiae and femora which could be dissected quickly to avoid post-mortem migration. For the rest of the skeleton, the effect of the latter was minimized by allowing recirculation for 5 min and comparing blood flow determinations in conjunction with a cardiac output measurement. The apparent extraction ratio of18F was variable and always exceeded unity in the single-passage experiments. With recirculation, there were significant differences between the extraction ratios for some bones, and the ratios sometimes exceeded unity. Extraction ratios for85Sr were similarly variable, but lower. When85Sr was compared directly with18F, the relative extraction ratio was 0.8, with little variation between bones or animals. It is concluded that the flux of microspheres entering the vasculature of bone is not representative of blood flow, owing to the removal of some microspheres in pre-osseous capillary beds. Microspheres are therefore not a satisfactory standard for the determination of extraction ratios.
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  • 92
    ISSN: 1432-0827
    Keywords: Ectopic calcification ; 1,25-dihydroxyvitamin D ; hyperphosphatemia ; Bone ; Renal tubule
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary A 50-year-old Latin American man with tumoral calcinosis presented with hyperphosphatemia (6.62±1.04 SD mg/dl), elevated renal threshold phosphorus concentration (TmP) (7.3 mg/GFR), and 1,25-dihydroxyvitamin D [1,25-(OH)2D] (69 pg/ml) hypercalciuria (239 mg/day), and a high fractional intestinal calcium (Ca) absorption (0.74). Sodium cellulose phosphate therapy (20 g/day) lowered urinary Ca, and partially reduced serum phosphorus (P) and TmP to 5.91±0.63 mg/dl and 6.2 mg/GFR, respectively. Serum 1,25-(OH)120D remained elevated at 58–64 pg/ml. Amphojel therapy (4 oz/day) decreased urinary P to 23±21 mg/day and lowered serum P to 5.75±0.36 mg/dl (P〈0.05). TmP increased to a value of 8.0 mg/GFR while serum 1,25-(OH)2D continued to remain elevated at 53 pg/ml. This case illustrates the probable operation of dual abnormalities in tumoral calcinosis represented by augmented renal conservation of P and an elevation in the circulating concentration of 1,25-(OH)2D.
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  • 93
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    Calcified tissue international 32 (1980), S. 69-76 
    ISSN: 1432-0827
    Keywords: Bone ; Marrow ; Blood flow ; Microspheres
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Blood flow was measured in mature and immature dogs by means of tracer microspheres. Microspheres of 15 µm were found to be the most suitable size in the dog. Total body nonentrapment in the awake, standing dog is likely to be less than 10%. Cortical bone flood flow, devoid of periosteum and marrow, is 2.5 ml/100 g/min in mature dogs and 7.0 ml/100 g/min in immature dogs,P〈0.005. Blood flow in cancellous bone is greater than that in cortical bone in mature (P〈0.001) and immature (P〈0.02) dogs. Flow is different in different regions of a long bone because of different proportions of cortical and cancellous bone, probably because of interrelationships of function (surfaces undergoing remodeling) and, therefore, of energy metabolism and blood flow.
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  • 94
    ISSN: 1432-0827
    Keywords: 1.25 (OH)2D3 ; Fish ; Bone ; Osteoblasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Female mature eels (300 to 500 g) received one intraperitoneal injection of 1.25 (OH)2D3 (10 µg). Their vertebral bone was compared, 8 h and 24 h after the injection, with vertebral bone of control mature female eels receiving solvent alone (ethanol). Sexual maturation in female eels induces a bone decalcification with hypercalcaemia and hyperphosphataemia. The control eels showed marked osteoclastic resorption and osteocytic osteolysis and the degree of mineralization of the intercellular substance decreased. Injection of 1.25 (OH)2D3 into these female mature eels provoked as early as 8 h : 1) an increase in hypercalcaemia and hyperphosphataemia ; 2) a major conversion of lining cells to osteoblasts and a stimulation of osteoblastic activity with new bone formation ; 3) diminished osteoclastic resorption without changing osteocytic osteolysis or bone matrix mineralization.
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  • 95
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    Calcified tissue international 32 (1980), S. 113-122 
    ISSN: 1432-0827
    Keywords: Rat ; Bone ; Metaphysis ; Quantitative ; Aging
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The purpose of this work was to analyze the proximal tibial metaphysis of the 170 g rat in a quantitative histologic fashion which would allow some relation to tissue age to be established. Stained 3 µm thick tissue sections were analyzed with the aid of a Merz grid on an eyepiece reticule and a light microscope. Tissue mass and cell distribution were studied in all areas. The rate of change in tissue mass during aging of the metaphysis was calculated. Two regions of the metaphysis were identified. One, corresponding to the primary spongiosa, less than 4.45 days of age, is a region of high turnover of hard tissue and high numbers of osteoblasts and osteoprogenitor cells. The other, corresponding to the secondary spongiosa, is a region of relatively low net tissue turnover and low numbers of osteoblasts and osteoprogenitor cells. Osteoclasts were found relatively more uniformly distributed through the metaphysis than were osteoblasts and osteoprogenitor cells. The rate of bone formation in the primary spongiosa is 50 times that found in the Haversian bone of the rib of 5-year-old humans and about 500 times that found at the cortical-endosteal surface of ribs of 5-year-old humans. It is argued that both cell distribution and tissue distribution in the metaphysis support the concept that osteoblasts and osteoclasts, rather than osteocytes, are responsible for the maturation of the metaphysis. The inhomogeneous distribution of both cells and tissue in the metaphysis has definite meaning for the interpretation of findings concerning the incorporation of radionuclides into the skeleton.
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  • 96
    ISSN: 1432-0827
    Keywords: Bone ; Mineral ; Crystallinity ; Maturation ; Age
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The crystallinity of bone mineral at different stages of maturation has been measured by quantitative X-ray diffraction methods. Crystallinity measurements were made on tibial middiaphyses from 17-day embryonic chicks, newlyformed periosteal bone from embryonic chicks, and density-fractionated bone from post-hatch chickens from 5 weeks to 2 years of age. For a given animal age and degree of mineralization, crystallinity increases with animal age, indicating that changes in bone mineral occur even after mineralization is complete or nearly complete.
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  • 97
    ISSN: 1432-0827
    Keywords: ADP ; Bisphosphonate ; Bone ; Resorption ; Formation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The effects of 1.5–2 years oral administration of disodium (3-amino-1-hydroxypropylidene)-1,1-bisphosphonate (APD) on bone metabolism were studied in male and female rats. APD was mixed in the food at levels of 500, 2,000 and 10,000 ppm. A dose-dependent increase in metaphyseal bone was found, indicative of continued inhibition of bone and cartilage resorption. APD did not affect mineralization of bone and cartilage, primary bone formation, or periosteal apposition. A short-term metabolic balance study was performed to compare the effects of oral with subcutaneous APD. Absorption of APD was in the order of 0.2%. Oral APD increased absorption of phosphate, probably by complexation of calcium with APD. The excess absorbed phosphate increased phosphaturia and decreased urinary calcium.
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  • 98
    ISSN: 1432-0827
    Keywords: Plasma proteins ; IgE ; IgD ; α 1Acid-glycoprotein ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Human cortical bones were extracted with EDTA, and the residue after EDTA extraction was digested with bacterial collagenase. Ten plasma proteins were identified and quantitated in the EDTA extracts. Three of them—IgE, IgD, andα 1acid-glycoprotein—had not previously been described in bone or dentine. Five plasma proteins identified in collagenase digests are albumin, IgG, IgA, IgE, andα 1acid-glycoprotein. IgE,α 1acid-glycoprotein, andα 2HS-glycoprotein were found to be concentrated in the bone more than other plasma proteins by factors between 11 and 525. The identification of plasma proteins was facilitated by the addition of polyethylene glycol in agarose gel. The presence of plasma proteins both in EDTA extracts and in collagenase digests suggests their structural role in bone.
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  • 99
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    Calcified tissue international 34 (1982), S. 253-257 
    ISSN: 1432-0827
    Keywords: Prednisolone ; Calcium ; Bone ; Corticoid osteopenia ; Vitamin D metabolites
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Adult male rats were fed a diet containing 0.15% calcium, 0.3% phosphorus, and either 100, 50, or 20 mg of prednisolone per kg of diet. All these levels of prednisolone led to osteopenia, decreased intestinal absorption of calcium, slightly lower serum calcium and phosphorus, and a decreased level of serum 1,25-dihydroxyvitamin D3. Exogenous parenteral 1,25-dihydroxyvitamin D3 corrected steroid-induced changes in serum calcium and phosphorus, but could not completely correct the low intestinal calcium transport; nor did it prevent the development of osteopenia. The prednisolone-induced osteopenia seems at least in part to be caused by impaired intestinal calcium transport. The impaired calcium transport may be the result of low levels of 1,25-dihydroxyvitamin D3 and a direct effect of presnisolone on the intestine.
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  • 100
    ISSN: 1432-0827
    Keywords: Vitamin D ; Vitamin D deficiency ; Bone ; Pregnancy ; Lactation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The effect of vitamin D on bone changes during the reproductive cycle in female rats has been investigated. One group of female rats was maintained on a vitamin D-deficient diet and another group on a vitamin D-replete diet from weaning. Both groups were mated with normal males and changes in their bones were determined histomorphometrically during pregnancy, lactation, and after weaning. All vitamin D-deficient rats had bone changes typical of rickets. Pregnancy caused significant reductions in mineralized tissue of trabecular and cortical bone in the vitamin D-deficient rats. Lactation caused further significant reductions in mineralized tissues of cortical and trabecular bone in both the vitamin D-deficient and vitamin D-replete animals, with the greatest changes seen at weaning. Some restoration of mineralized tissues occurred following weaning. There was an increase in tetracycline-labeled bone surface in the vitamin D-replete animals during lactation, likely due to an increase in bone formation rates. In the vitamin D-deficient animals during lactation, there was a decrease in tetracyclinelabeled bone surface, likely due to severely depressed bone mineralization. These results indicate that the mobilization of calcium from bone to maintain pregnancy and lactation occurs by a mechanism independent of vitamin D.
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