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  • Nitrogen fixation  (148)
  • Bone  (145)
  • Springer  (293)
  • Cambridge University Press
  • 1980-1984  (191)
  • 1975-1979  (102)
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  • 1
    ISSN: 1432-2048
    Keywords: Glyoxylate ; Isonicotinic acid hydracide ; Medicago ; Nitrogen fixation ; Photorespiration ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nitrogen fixation (measured as acetylene reduction) by whole nodulated alfalfa plants was stimulated when the plants were treated with isonicotinic acid hydracide (INH) and glyoxylate, both inhibitors of the glycolate pathway of carbohydrate metabolism, at concentrations of 300 and 100 mM, respectively. Reducing energetic loses caused by photorespiration results in an increase in the symbiotic nitrogen fixation.
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  • 2
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    Planta 152 (1981), S. 544-552 
    ISSN: 1432-2048
    Keywords: Ammonium assimilation ; Lichens ; Nitrogen fixation ; Peltigera
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Nostoc in the cephalodia of the lichen Peltigera aphthosa Willd. fixed 15N2 and the bulk of the nitrogen fixed was continuously transferred from it to its eukaryotic partners (a fungus and a green alga, Coccomyxa sp.). Kinetic studies carried out over the first 30 min, after exposure of isolated cephalodia to 15N2, showed that highest initial 15N2-labelling was into NH 4 + . After 12 min little further increase in the NH 4 + label occurred while that in the amide group of glutamine and in glutamate continued to increase. The 15N-labelling of the amino group of glutamine and of aspartate increased more slowly, followed by an increase in the labelling of alanine. When total incorporation of 15N-label was calculated, the overall pattern was found to be rather similar except that, throughout the experiment, the total 15N incorporated into glutamate was about six times greater than that into the amide group of glutamine. Pulse chase experiments, in which 14N2 was added to cephalodia previously exposed to 15N2, showed that the NH 4 + pool rapidly became depleted of 15N-label, followed by decreases in the labelling of glutamate, the amide group of glutamine and aspartate. The 15N-labelling of alanine, however, continued to increase for a period. When isolated cephalodia were treated with L-methionine-SR-sulphoximine, an inhibitor of glutamine synthetase (EC 6.3.1.2), and azaserine, an inhibitor of glutamate synthase (EC 2.6.1.53), there was no detectable labelling in glutamine although the 15N-labelling of glutamate increased unimpaired. On treating the cephalodia with amino-oxyacetate, an inhibitor of aminotransferase activity, the alanine pool decreased. Evidence was obtained that glutamine synthetase and glutamate synthase were located in the Nostoc, and that glutamate dehydrogenase (EC 1.4.1.4) and various amino-transferases were located in the cephalodial fungus. Possible implications of these findings are discussed.
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  • 3
    ISSN: 1432-2048
    Keywords: Ammonia ; Nitrogen fixation ; Nodule ; Senescence (root nodules) ; Ureide ; Vigna
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract During early development (up to 18 d after sowing) of nodules of an “effective” cowpea symbiosis (Vigna unguiculata (L.) Walp cv. Vita 3: Rhizobium strain CB756), rapidly increasing nitrogenase (EC 1.7.99.2) activity and leghaemoglobin content were accompanied by rapid increases in activities of glutamine synthetase (EC 6.3.1.2), glutamate synthase (EC 2.6.1.53), enzymes of denovo purine synthesis (forming inosine monophosphate) xanthine oxidoreductase (EC 1.2.3.2), urate oxidase (EC 1.7.3.3), phosphoenolpyruvate carboxylase (EC 4.1.1.31) and led to increased export of ureides (allantoin and allantoic acid) to the shoot of the host plant in the xylem. Culturing plants with the nodulated root systems maintained in the absence of N2 (in 80 Ar: 20 O2, v/v) had little effect on the rates of induction and increase in nitrogenase activity and leghaemoglobin content but, in the absence of N2 fixation and consequent ammonia production by bacteroids, there was no stimulation of activity of enzymes of ammonia assimilation or of the synthesis of purines or ureides. Addition of NO 3 - (0.1–0.2 mM) relieved host-plant nitrogen deficiency caused by the Ar: O2 treatment but failed to increase levels of enzymes of N metabolism in either the bacteroid or the plant-cell fractions of the nodule. Premature senescence in Ar: O2-grown nodules occurred at 18–20 d after sowing, and resulted in reduced levels of nitrogenase activity and leghaemoglobin but increased the activity of hydroxybutyrate oxidoreductase (EC 1.1.1.30).
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  • 4
    ISSN: 1432-2048
    Keywords: Alnus ; Ammonium ; Carbon translocation ; Endophyte damage ; Nitrogen fixation ; Root nodule
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cloned plants of Alnus incana (L.) Moench were inoculated and grown without combined nitrogen for seven weeks. The effects of ammonium on the function and structure of the root nodules were studied by adding 20 mM NH4Cl (20 mM KCl=control) for four days. Nitrogenase activity decreased to ca. 50% after one day and to less than 10% after two days in ammonium treated plants, but was unaffected in control plants. The results were similar at photon flux densities of 200 and 50 μmol m-2 s-1. At the higher light level the effect was concentration dependent between 2 and 20 mM NH4Cl. The recovery was slow, and more than 11 d were needed for plants treated with 20 mM ammonium to reach initial activity. The distribution of 14C to the root nodules after assimilation of 14CO2 by the plants was not changed by the ammonium treatment. Microscopical studies of root nodules showed high frequencies of endophyte vesicles being visually damaged in nodules from ammonium-treated plants, but not in nodules from control plants. When nitrogenase activity was restored, visually damaged vesicles were again few, whereas young developing vesicles were numerous. The slow recovery, the 14C-translocation pattern, and the structural changes of the endophyte indicate a more complex mechanism of ammonium influence than simply a short-term reduction in supply of carbon compounds to the nodules.
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  • 5
    ISSN: 1432-2048
    Keywords: Allantoin ; Amino acids ; Bleeding sap ; Nitrogen fixation ; Phaseolus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Dwarf french beans, Phaseolus vulgaris L., were grown with or without inoculation with rhizobia (strain 3644), and with or without a combined nitrogen source (nitrate or ammonium ions). The distribution of radioactivity into products of dark 14CO2 assimilation was studied in roots or nodules from these plants. A detailed study was also made of the distribution and rates of excretion of nitrogen in xylem bleeding sap in 28 day old plants grown on the various sources of nitrogen. Whereas detached nodules accumulated radioactive glycine, serine and glutamate when incubated with 14CO2, bleeding sap from plants root fed 14CO2 contained low levels of radioactivity in these compounds but higher levels in allantoin. Chemical analysis showed allantoin to be the major compound transported in the xylem of nodulated plants, whether or not they were fed on combined nitrogen. In contrast uninoculated plants accumulated mainly amino acids in the bleeding sap, the amount and chemical composition of which depended on the combined nitrogen source.
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  • 6
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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
    Source: Springer Online Journal Archives 1860-2000
    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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  • 7
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    Calcified tissue international 33 (1981), S. 35-39 
    ISSN: 1432-0827
    Keywords: Copper ; Resorption ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary An investigation of the role of copper in bone metabolism was undertaken. Explanted calvaria from 6-day-old mice were grown for 48 h in medium with and without the addition of copper sulfate. Active resorption was found to be significantly inhibited in the presence of copper sulfate concentrations of 10−6M and above. Copper sulfate concentrations of 10−5M and above inhibited hydroxyproline, protein, and DNA synthesis. Lower concentrations were ineffective. The effect of 5 × 10−6M copper sulfate on resorption was reversible. Several other compounds were tested for similar effects and at 5 × 10−6M were found to inhibit bone resorption in the order: copper sulfate 〉 brown gold chloride 〉 sodium aurothiomalate 〉 zinc sulfate 〉 sodium sulfate. The copper sulfate effect was twice that of sodium aurothiomalate, and sodium sulfate was not significantly inhibitory. The results suggest that the high serum copper levels associated with rheumatoid arthritis may reflect the activity of a hypothetical control mechanism of bone resorption. In the diseased state this would act to restore the normal rate of bone resorption.
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  • 8
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    Calcified tissue international 22 (1977), S. 27-33 
    ISSN: 1432-0827
    Keywords: Protein ; Bone ; Blood plasma ; Dentine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract The spectrum of plasma proteins present in human cortical bone and permanent dentine has been determined. One plasma glycoprotein, theαHS-glycoprotein, was found to be present at a high concentration in both bone and dentine and was shown to be concentrated in the mineralized tissues with respect to the other plasma proteins by factors of between 30 and 100. In this respect theαHS-glycoprotein is analogous to the G2B-glycoprotein and α-glycoprotein of bovine and rabbit b one, respectively. The binding ofαHS-glycoprotein and albumin to calcium phosphates generated within serum samples has been studied at different serum: precipitate ratios. In each case all theαHS-glycoprotein was removed from the samples and theαHS-glycoprotein was concentrated with respect to albumin by factors ranging from 370 at the highest serum: precipitate ratio to 25 at the lowest ratio. The plasmaαHS-glycoprotein concentrations of patients with Paget's disease of bone were shown to be substantially lower than the normal range, with significant negative correlation between theαHS-glycoprotein concentration and the plasma alkaline phosphatase activity.
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  • 9
    ISSN: 1432-0827
    Keywords: Bone ; Calcium ; Strontium ; Apposition ; Tracer analysis
    Source: Springer Online Journal Archives 1860-2000
    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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  • 10
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    Calcified tissue international 25 (1978), S. 29-35 
    ISSN: 1432-0827
    Keywords: Albumin ; Bone ; Growth retardation ; Osteomalacia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary 125I-labeled rat albumin injected intravenously into rats was taken up by growing bone. Some of this radioactive albumin could be removed from bone by washing with saline, the proportion so removed decreasing from 82.5% at 1 day to 7.4% at 8 days. Both the total radioactivity, and that remaining in bone after saline wash, were reduced when growth was slowed by alteration or restriction of the diet. Although the amount of125I albumin in rachitic bone was reduced, autoradiography showed that radioactivity was present in rachitic osteoid. Immunoprecipitation using anti-rat-albumin serum showed that about half of the radioactivity released from normal bone by EDTA was still attached to albumin. These results suggest that albumin plays some part in the growth of bone.
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  • 11
    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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  • 12
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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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  • 13
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    Calcified tissue international 27 (1979), S. 33-40 
    ISSN: 1432-0827
    Keywords: Chick embryo ; Bone ; Organ culture ; Scanning electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    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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  • 14
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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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  • 15
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    Calcified tissue international 23 (1977), S. 215-223 
    ISSN: 1432-0827
    Keywords: Amorphous mineral ; Bone ; Electron microscopy ; Ultracryotomy ; Ultramicro-incineration
    Source: Springer Online Journal Archives 1860-2000
    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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  • 16
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    Calcified tissue international 26 (1978), S. 85-89 
    ISSN: 1432-0827
    Keywords: Bone ; Histology ; Microradiography ; Densitometry ; Computer
    Source: Springer Online Journal Archives 1860-2000
    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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  • 17
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    Calcified tissue international 26 (1978), S. 139-142 
    ISSN: 1432-0827
    Keywords: Chemisorption ; Surface area ; Hydroxyapatite ; Bone ; Enamel
    Source: Springer Online Journal Archives 1860-2000
    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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  • 18
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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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  • 19
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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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  • 20
    ISSN: 1432-0827
    Keywords: Bone ; Bone cells ; Cyclic AMP ; Osteoclast activating factor ; Lymphokine
    Source: Springer Online Journal Archives 1860-2000
    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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  • 21
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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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  • 22
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    Calcified tissue international 27 (1979), S. 233-237 
    ISSN: 1432-0827
    Keywords: Bone ; Vitamin D ; Acidosis ; Phosphate
    Source: Springer Online Journal Archives 1860-2000
    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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  • 23
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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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  • 24
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    Calcified tissue international 26 (1978), S. 209-213 
    ISSN: 1432-0827
    Keywords: Mithramycin ; Bone ; β-Glucuronidase ; KT medium
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    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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  • 25
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    Calcified tissue international 29 (1979), S. 169-171 
    ISSN: 1432-0827
    Keywords: Glycosaminoglycans ; Bone
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    Topics: Biology , Medicine , Physics
    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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  • 26
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    Calcified tissue international 31 (1980), S. 135-145 
    ISSN: 1432-0827
    Keywords: Bone ; water compartments ; marker ; methods ; pH ; potassium ; lactate
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    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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  • 27
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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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  • 28
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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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    Topics: Biology , Medicine , Physics
    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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  • 29
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    Calcified tissue international 31 (1980), S. 261-265 
    ISSN: 1432-0827
    Keywords: Bone ; blood flow ; microspheres ; fluorine ; strontium
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    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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  • 30
    ISSN: 1432-0827
    Keywords: Ectopic calcification ; 1,25-dihydroxyvitamin D ; hyperphosphatemia ; Bone ; Renal tubule
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    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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  • 31
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    Calcified tissue international 32 (1980), S. 69-76 
    ISSN: 1432-0827
    Keywords: Bone ; Marrow ; Blood flow ; Microspheres
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    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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  • 32
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    Keywords: 1.25 (OH)2D3 ; Fish ; Bone ; Osteoblasts
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    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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  • 33
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    Calcified tissue international 32 (1980), S. 113-122 
    ISSN: 1432-0827
    Keywords: Rat ; Bone ; Metaphysis ; Quantitative ; Aging
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    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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  • 34
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    Calcified tissue international 32 (1980), S. 123-133 
    ISSN: 1432-0827
    Keywords: Bone ; Cell kinetics ; Osteoprogenitor cell ; Migration ; Differentiation
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    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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  • 35
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    Keywords: APD ; Bisphosphonate ; Bone ; Resorption ; Formation
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    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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  • 36
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    Keywords: Bone ; Calcium metabolism ; Whole-body counting
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    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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  • 37
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    Calcified tissue international 34 (1982), S. 103-110 
    ISSN: 1432-0827
    Keywords: Bone ; Collagen ; Vitamin D metabolites
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    Topics: Biology , Medicine , Physics
    Notes: Summary The effects of selected vitamin D3 metabolites and analogs on bone collagen synthesis in vitro were examined in organ cultures of neonatal mouse calvarial bone. The incorporation of [3H]proline into the collagenase-digestible fraction of newly synthesized protein was progressively inhibited by 1α,25-dihydroxyvitamin D3 (1α,25-(OH)2D3) (10−12 M to 10−7 M) in 24-h cultures, and incorporation into noncollagen protein was also blunted at the higher doses employed. The synthetic analog 1α-hydroxyvitamin D3 (1α-OHD3) was almost 300-fold less potent an inhibitor of collagen synthesis than was 1α,25(OH)2D3, and the natural metabolites 25-hydroxyvitamin D3 (25OHD3) and 24R,25-dihydroxyvitamin D3 (24R,25(OH)2D3), 1000-fold less potent, although the dose-response curve for each of these compounds was not parallel with that for 1α,25(OH)2D3. The 24S,25(OH)2D3 enantiomer was four-fold less potent than 24R,25-(OH)2D3 or 25OHD3, and vitamin D3 showed less than 2% the activity of 25OHD3. The responses were unaffected by the substitution of 0.4% bovine albumin for 5% horse serum in the medium, and no stimulation of collagen synthesis was observed in response to 25-hydroxylated metabolites between 2×10−14 and 2×10−6 M or in cultures treated for up to 96 h with 24R,25(OH)2D3 (2×10−10M). The overall results emphasize the similarity of the structural requirements for the inhibition of matrix synthesis and the stimulation of resorption by active vitamin D metabolites in bone. In addition, these studies support the importance of the 1-hydroxyl function to the biologic activity of vitamin D in the skeleton.
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  • 38
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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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  • 39
    ISSN: 1432-0827
    Keywords: Plasma proteins ; IgE ; IgD ; α 1Acid-glycoprotein ; Bone
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    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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  • 40
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    Calcified tissue international 34 (1982), S. 253-257 
    ISSN: 1432-0827
    Keywords: Prednisolone ; Calcium ; Bone ; Corticoid osteopenia ; Vitamin D metabolites
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    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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  • 41
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    Keywords: Vitamin D ; Vitamin D deficiency ; Bone ; Pregnancy ; Lactation
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    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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  • 42
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    Calcified tissue international 36 (1984), S. S25 
    ISSN: 1432-0827
    Keywords: Remodeling ; Bone ; Stress
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    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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  • 43
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    Calcified tissue international 34 (1982), S. 376-381 
    ISSN: 1432-0827
    Keywords: Matrix vesicles ; Bone ; Actin ; Rat
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    Topics: Biology , Medicine , Physics
    Notes: Summary Preliminary indications of the occurrence of actin and myosin in crude matrix vesicle preparations have been reported previously. In the present study extracellular matrix vesicles from rat alveolar bone were isolated. They were further purified by a sucrose density gradient. SDS-polyacrylamide gel electrophoresis of the purified vesicles revealed the presence of a polypeptide with a molecular weight of 43 K daltons and with electrophoretic mobility identical to that of blood platelet actin. The limited proteolysis of both 43 K dalton vesicular polypeptide and actin byStaphylococcus aureus-V8-protease revealed three fragments with identical electrophoretic mobility. In addition, the vesicular preparations inhibited the activity of DNase I, a property typical of actin monomers. Filamentous material extracted from matrix vesicles showed ultrastructural features of F-actin. Reaction of this material with heavy meromyosin resulted in arrowhead formation, which is characteristic of acto-heavy meromyosin. The occurrence of actin in extracellular matrix vesicles may account for their budding from the osteoblastic plasma membrane, their possible motility in the matrix, and maintenance of the spherical shape.
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  • 44
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    Calcified tissue international 35 (1983), S. 148-152 
    ISSN: 1432-0827
    Keywords: Bone ; Computed tomography ; Densitometry ; Osteoporosis ; Osteomalacia
    Source: Springer Online Journal Archives 1860-2000
    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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  • 45
    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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  • 46
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    Calcified tissue international 33 (1981), S. 57-69 
    ISSN: 1432-0827
    Keywords: Thallium ; Chondrodystrophy ; Cartilage ; Bone ; Matrix
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    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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  • 47
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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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  • 48
    ISSN: 1432-0827
    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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  • 49
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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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  • 50
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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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  • 51
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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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  • 52
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    Calcified tissue international 33 (1981), S. 631-639 
    ISSN: 1432-0827
    Keywords: Bone ; Osteopenia ; Noninvasive monitoring
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    Topics: Biology , Medicine , Physics
    Notes: Summary Radiological techniques were utilized for monitoring progressive changes in compact bone in the tibia of monkeys during experimentally induced osteopenia. Bone mass loss in the tibia during restraint was evaluated from radiographs, from bone mineral analysis, and from images reconstructed from gamma ray computerized tomography. The losses during 6 months of restraint tended to occur predominantly in the proximal tibia and were characterized by subperiosteal bone loss, intracortical striations, and scalloped endosteal surfaces. Bone mineral content in the cross section of the tibia declined 17–21%. Tomography demonstrated endosteal widening and reduced mineral content per unit of thickness of cortical bone. In 6 months of recovery, the mineral content of the proximal tibia remained depressed. Effects of the dynamic environment on local-regional changes in various skeletal areas are discussed.
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  • 53
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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
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    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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  • 54
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    Calcified tissue international 22 (1977), S. 183-189 
    ISSN: 1432-0827
    Keywords: Calcium ; Strontium ; Bone ; Mathematical modelling
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    Topics: Biology , Medicine , Physics
    Notes: Abstract 1. A method has been devised for comparing the impulse response functions of the skeleton for two or more boneseeking tracers, and for estimating the contribution made by measurement errors to the differences between any pair of impulse response functions. 2. Comparisons were made between the calculated impulse response functions for47Ca and85Sr obtained in simultaneous double tracer studies in sixteen subjects. Collectively the differences between the47Ca and85Sr functions could be accounted for entirely by measurement errors. 3. Because the calculation of an impulse response function requires fewera priori assumptions than other forms of mathematical analysis, and automatically corrects for differences induced by recycling of tracer and non-identical rates of excretory plasma clearance of tracer, it is concluded that differences shown in previous in vivo studies between the fluxes of*Ca and*Sr into bone can be fully accounted for by undetermined oversimplifications in the various mathematical models used to analyse the results of those studies.85Sr is therefore an adequate tracer for bone calcium in most in vivo studies.
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  • 55
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    Calcified tissue international 22 (1977), S. 219-226 
    ISSN: 1432-0827
    Keywords: Hydroxyapatite ; Bone ; X-ray Diffraction
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    Notes: Abstract Evidence relating to the existence of crystalline bone mineral in vivo is considered, and bone apatite crystal structure investigated using an x-ray powder diffraction technique. Specimens of femoral compacta excised post-mortem from male and female subjects ranging from 3 1/2 years to 87 years of age have been studied. Values of the ratio c/a of bone apatite crystal cell axes for females are significantly higher (p〈0.05) than for males. Moreover, significant change of c/a with age is observed for males (p=0.0005) but not for females (p=0.30). Differences in c/a are interpreted as indicating substitution of constituent ions in the bone apatite crystals.
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  • 56
    ISSN: 1432-0827
    Keywords: Diphosphonate ; Bone ; Osteoclasts ; Autoradiography
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    Notes: Abstract Male rats weighing 100 g were injected with vehicle (control group), 0.4 or 4.0 mg/kg/day of ethane-1-hydroxy-1,1-diphosphonate (EHDP). Beginning after 6 days of EHDP treatment all rats received six subcutaneous injections of3H-thymidine at 8 h intervals. The multiple labeling index of autoradiographically defined labeled cells was determined for the osteoprogenitor cells of the primary spongiosa of the proximal tibial metaphysis. The osteoclasts in the proximal tibial metaphysis were assayed for population parameters and incorporation of labeled nuclei at sequential intervals after the administration of3H-thymidine. After 6 days of EHDP pre-treatment the multiple labeling index of the osteoprogenitor cells was reduced with 4.0 mg/kg/day dose of EHDP. At the beginning of the experimental kinetic sampling period, after the 6 days of EHDP pre-treatment, EHDP treatment resulted in significant increases in the numbers of osteoclasts, nuclei per osteoclast profile, and total osteoclast nuclei. During the sampling period, the 4.0 mg/kg/day dose of EHDP caused further increases in the number of total osteoclasts. EHDP treatment caused a marked increase in the rate of incorporation and accumulation of3H-thymidine-labeled nuclei in osteoclasts. Thus, EHDP causes an accelerated rate of osteoclast production from precursor cells in spite of its ability to slow bone resorption. Although the origin of osteoclast nuclei is not clear, the kinetic data from this study suggests that osteoclast nuclei are not arising from osteoprogenitor cells or osteocytes in the EHDP-treated animals.
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  • 57
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    Calcified tissue international 22 (1977), S. 303-313 
    ISSN: 1432-0827
    Keywords: Osteoclasts ; Sedimentation ; Isolation ; Cells ; Bone
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    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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  • 58
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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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  • 59
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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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  • 60
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    Calcified tissue international 22 (1977), S. 329-331 
    ISSN: 1432-0827
    Keywords: Bone ; Fracture ; Mineral
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    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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  • 61
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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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  • 62
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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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  • 63
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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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    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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  • 64
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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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  • 65
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    Calcified tissue international 23 (1977), S. 161-170 
    ISSN: 1432-0827
    Keywords: Bone ; Growth ; Rat ; Remodeling ; Tetracyclines
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    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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    Calcified tissue international 23 (1977), S. 1-5 
    ISSN: 1432-0827
    Keywords: Water ; Bone ; Homeostasis ; Structure
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    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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    Calcified tissue international 27 (1979), S. 177-185 
    ISSN: 1432-0827
    Keywords: Calcification ; Bone ; Glycoprotein ; Golgi ; Osteoblasts
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    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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  • 68
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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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  • 69
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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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  • 70
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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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  • 71
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    Calcified tissue international 25 (1978), S. 105-110 
    ISSN: 1432-0827
    Keywords: Progesterone ; 17β estradiol ; Testosterone ; Tissue culture ; Bone
    Source: Springer Online Journal Archives 1860-2000
    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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  • 72
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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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  • 73
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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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  • 74
    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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  • 75
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    Calcified tissue international 26 (1978), S. 81-84 
    ISSN: 1432-0827
    Keywords: Osteoclasts ; Bone ; Resorption
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    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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  • 76
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    Calcified tissue international 30 (1980), S. 67-71 
    ISSN: 1432-0827
    Keywords: Alkaline phosphatase ; ATPase ; Bone ; Triton
    Source: Springer Online Journal Archives 1860-2000
    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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  • 77
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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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  • 78
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    Keywords: Vitamin D ; Bone ; Osteoblasts ; Receptors ; 1,25-dihydroxyvitamin D3
    Source: Springer Online Journal Archives 1860-2000
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    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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  • 79
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    Keywords: Bone ; Mineral ; Crystallinity ; Maturation ; Age
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    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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  • 80
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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
    Source: Springer Online Journal Archives 1860-2000
    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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  • 82
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    Keywords: Vitamin D ; Vitamin D deficiency ; Bone ; Cartilage ; Bone development
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    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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  • 83
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    Calcified tissue international 35 (1983), S. 728-731 
    ISSN: 1432-0827
    Keywords: Bone ; Bioelectricity
    Source: Springer Online Journal Archives 1860-2000
    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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    Calcified tissue international 34 (1982), S. 125-130 
    ISSN: 1432-0827
    Keywords: X-Linked hypophosphatemic rickets ; Osteomalacia ; 1,25-Dihydroxyvitamin D ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary In order to determine whether a defect in vitamin D metabolism might play a role in the pathogenesis of X-linked hypophosphatemic rickets and osteomalacia (XLH), we compared the serum 1,25-dihydroxyvitamin D [1,25(OH)2D] level in 52 normal subjects and 37 patients with XLH. In untreated patients, adults were found to have values similar to age-matched controls, while youths had values similar to growth-rate-matched controls but significantly lower than the levels of age-matched controls who were growing at a normal rate. In contrast, treated XLH patients of all ages had serum levels significantly lower than both controls and untreated XLH patients. Further, the serum levels of 1,25(OH)2D in these treated patients had a significant inverse linear correlation with serum 25-(OH)D concentrations. We propose that subjects with XLH have serum 1,25(OH)2D levels within appropriate age- and growth-rate-matched normal ranges. However, in the presence of hypophosphatemia, we would have anticipated elevated levels of 1,25(OH)2D; viewed in this light the serum 1,25(OH)2D levels are inadequate, suggesting the presence of a relative deficiency of this active vitamin D metabolite.
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    Calcified tissue international 34 (1982), S. 145-148 
    ISSN: 1432-0827
    Keywords: Bone ; Diphosphonate ; Phosphate fluxes ; Solubility
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Calvaria taken from rats (normal or thyroparathyroidectomized) given a diphosphonate (EHDP 10 mg/kg daily for 7 days) were placed in special Ussing chambers for the measurement of phosphate fluxes to and from bone. When compared with appropriate controls, the experimental calvaria showed a marked decrease in influx, K, a significant increase in the projected equilibrium concentration, E/K, where E is the efflux and a significnant increase in the calcium concentration in the medium. It is concluded that this large increase in passive “solubility” is caused by the diphosphonate's ability to stabilize the small amount of regulator phase (brushite or brushite-apatite mixture) present in bone.
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    Calcified tissue international 34 (1982), S. 370-375 
    ISSN: 1432-0827
    Keywords: Complement ; Bone ; Collagen ; Prostaglandins
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary Activation of rabbit serum complement caused a marked reduction in collagen synthesis but a much smaller change in noncollagen protein synthesis in fetal rat calvaria maintained in organ culture. In the periosteum of the fetal rat calvarium, both collagen and noncollagen protein synthesis were reduced, whereas in the central bone, presumably enriched in osteoblasts, only collagen synthesis was inhibited. This large decrease in bone collagen synthesis could not be attributed to enhanced degradation of newly synthesized collagen or its release into the culture medium. Activation of complement also stimulated the production of PGE in fetal rat calvaria. Antagonists of prostaglandin cyclooxygenase decreased prostaglandin synthesis but did not restore collagen synthesis in complement-treated bones, suggesting that complement decreases osteoblast collagen synthesis by a mechanism largely independent of prostaglandin production.
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    Calcified tissue international 34 (1982), S. 403-407 
    ISSN: 1432-0827
    Keywords: Cyclic AMP ; Parathyroid hormone ; Bone ; Chronic uremia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The release of cyclic AMP from bone in response to stimulation with PTH 1–34 was examined in 20 dogs with long-term chronic renal failure (CRF) produced by unilateral nephrectomy and contralateral partial renal artery ligation. After 9 to 15 months of uremia, the tibiae were removed and perfused in vitro. Seven dogs with CRF served as controls, 7 dogs with CRF were treated with 24,25(OH)2D3 — 2.5 µg per day, and 6 CRF dogs underwent thyroparathyroidectomy (TPTX) 42 h before they were sacrificed. The release of cyclic AMP from bone in response to PTH 1–34 in the CRF dogs was severely reduced compared to the response observed in 7 dogs with normal renal function (net accumulation of cyclic AMP release 86±8.5 versus 426±59.0 pmol/30 min). Long-term treatment of uremic dogs with 24,25(OH)2D3 had no effect on the release of cyclic AMP by bone. However, the release of cyclic AMP was restored to normal levels in the CRF dogs that underwent thyroparathyroidectomy. All CRF dogs had secondary hyperparathyroidism and the fact that TPTX returned the cyclic AMP response to normal values suggests that desensitization to PTH of the adenylate cyclase system of bone exists in chronic uremia.
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    Calcified tissue international 34 (1982), S. 439-448 
    ISSN: 1432-0827
    Keywords: Bone ; Histomorphometry ; Osteocytes ; Bone dynamics ; Histology
    Source: Springer Online Journal Archives 1860-2000
    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
    Source: Springer Online Journal Archives 1860-2000
    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 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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  • 91
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    Calcified tissue international 33 (1981), S. 369-374 
    ISSN: 1432-0827
    Keywords: Histomorphometry ; Bone ; Reproducibility
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary To study bone histomorphometry reproducibility in normal subjects, we performed during orthopedic surgery bone biopsies in 16 post-menopausal women. Each woman had four bone biopsies, two at the usual site in the iliac crest, one on the left and one on the right side, and two other biopsies just behind the usual site, one at each side. We performed measurements of trabecular bone volume, relative osteoid volume, osteoid surfaces, osteoclastic resorption surfaces and calcification front. The average values of the 16 patients were compared, on the one hand, two by two, by a student test, and on the other hand, by a variance analysis. By these two methods the results showed no significant difference between the average values of the 16 patients at each location for any of the histomorphometric parameters studied. However, there was a location variation which was estimated by the intra-individual variation for a given patient. On the other hand, we calculated from the variance analysis the location variance for a group of 10 to 100 patients. In any case all the parameters had a location variation which was high for osteoclastic resorption surfaces and relative osteoid volume when expressed in % of the absolute value of these parameters. The variation of the trabecular bone volume was 0–46. 15% (95% confident limit interval) in a single patient and the hypothetical value of the location variation was 41.6% for a group of 10 patients and 13.0% for a group of 100 patients.
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  • 92
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    Calcified tissue international 33 (1981), S. 499-504 
    ISSN: 1432-0827
    Keywords: Bone ; Collagen ; Density
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The density of a bovine cortical bone matrix sample was found in water, several ethanol-water solutions, and in the dried state. Previously the density of the same mineralized bone was found fresh and when desiccated. The volume in each state was estimated from the dimensional changes axially, tangentially, and radially. Confirmation was found by determining the density of dried specimens upon immersion in xylene. The amount of imbibed xylene provided an estimate of the free pore volume in the dried matrix. The volume fraction of the solid constituent, S, in the wet matrix was found to be 0.57, from which the density of S in various solutions was calculated. Density of wet matrix in 0.15 M saline: 1.180 g/cc; for dried matrix, 1.246 g/cc. Density of wet S in saline: 1.33 g/cc; for dried S, 1.42 g/cc, which matches published values for collagen molecules. Dimensional changes between wet and dried state of matrix match published values for artificially cross-linked rat tail tendon fibers. Axially: 1.04, by area: 2.27; by volume: 2.62. Estimate of intrafibrillar volume, assuming 80% of mineral is within fibrils: 0.73 cc/g dry collagen.
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  • 93
    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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  • 94
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    Calcified tissue international 23 (1977), S. 103-112 
    ISSN: 1432-0827
    Keywords: Albumin ; Polyvinylpyrrolidone ; Tissue fluid ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The passage of tissue fluid through cortical bone has been investigated using radioactively labelled macromolecules as markers. The results suggest that in the cortex of young rabbit femur the movement of tissue fluid is in the same net direction as blood, mainly from the endosteal to the periosteal surface. Some albumin is incorporated from extravascular tissue fluid into calcified matrix at sites of bone formation. Polyvinylpyrrolidone, average molecular weight 35,000 is able to pass through extravascular tissue fluid in bone but isnot incorporated into calcified matrix. In rabbits made vitamin D deficient, much less albumin is retained in regions of bone formation than is the case with controls. Albumin adsorbs to the surface of calcium phosphate precipitates and it is suggested that this mechanism may be mainly responsible for its incorporation into bone.
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  • 95
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    Calcified tissue international 23 (1977), S. 185-188 
    ISSN: 1432-0827
    Keywords: Matrix vesicles ; Bone ; Ultracrytomy
    Source: Springer Online Journal Archives 1860-2000
    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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  • 96
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    Calcified tissue international 24 (1977), S. 93-98 
    ISSN: 1432-0827
    Keywords: Magnesium ; Bone ; Mineral ; Chick embryo ; Organ culture
    Source: Springer Online Journal Archives 1860-2000
    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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  • 97
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    Calcified tissue international 22 (1977), S. 9-17 
    ISSN: 1432-0827
    Keywords: Bone ; Fluoride ; Calcium ; Phosphorus ; Mechanical Strength
    Source: Springer Online Journal Archives 1860-2000
    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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  • 98
    ISSN: 1432-0827
    Keywords: Parathyroid hormone ; Plasma calcium ; Radiocalcium ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    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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  • 99
    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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  • 100
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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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