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  • Ultrastructure  (758)
  • Nitrogen fixation  (148)
  • Calcium  (143)
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  • American Geophysical Union (AGU)
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Monatshefte für Chemie 112 (1981), S. 149-156 
    ISSN: 1434-4475
    Keywords: Calcium ; Gallium ; Oxygen ; Single Crystal ; X-Ray
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract The hitherto unknown compound Ca3Ga4O9 was prepared and investigated by X-ray single crystal methods. Ca3Ga4O9 has orthorhombic symmetry:a=1435.8;b=1682.5;c=532.1 pm; space group C 2v 11 −Cmm2,Z=6. The tetrahedra network (circles of 4 and 5 GaO4-tetrahedra) and the surrounding of Ca2+ are described and discussed with respect to other oxogallates.
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  • 2
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    Calcified tissue international 33 (1981), S. 409-416 
    ISSN: 1432-0827
    Keywords: Bone Cells ; Cyclic AMP ; Calcium ; Parathyroid hormone ; Prostaglandin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary We have examined the influence of extracellular Ca2+ on cyclic AMP metabolism in an osteoblast-enriched population of bone cells isolated from the calvaria of rat fetuses. The cyclic AMP1 response to stimulators of cyclic AMP formation (PTH and PGE2), but not basal cyclic AMP levels, increased progressively as the extracellular Ca2+ concentration was raised from 0.2 to 4.0 mM. The response to changes in extracellular Ca2+ were rapid (within 3.5 min), and the level of responsivity that characterized each Ca2+ concentration persisted for at least 6 h when the Ca2+ concentration was kept constant. The effect of Ca2+ spanned the entire time course of PTH action, was not accompanied by altered excretion of cyclic AMP from the cells, and was evident at low as well as at high hormone concentrations. Ca2+ augmented the action of PTH in the presence as well as in the absence of cyclic AMP phosphodiesterase inhibitors, and failed to decrease cyclic AMP phosphodiesterase activity in the short term. Mn2+ and, to a smaller degree, Ba2+ substituted for Ca2+ in promoting the cyclic AMP response to PTH. Verapamil, an inhibitor of Ca2+ penetration, blunted the Ca2+-mediated increments in the cyclic AMP response, and the divalent cation ionophore A23187 enhanced these increments. These results indicate that Ca2+ and other cations are positive effectors of the stimulated cyclic AMP response in isolated bone cells. Accumulation into an as yet unknown cellular compartment may be required for the cation effect. The data are most consistent with enhancement of adenylate cyclase reactivity as the mode of cation action.
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  • 3
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    Calcified tissue international 33 (1981), S. 603-618 
    ISSN: 1432-0827
    Keywords: Preameloblasts ; Tooth germs ; Monkey ; Enamel ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Cytodifferentiation of inner enamel epithelium and the adjacent connective tissue from the tip of the cervical loop to the initiation of enamel elaboration in twoMacaca species was examined. Ten- to twelve-month-old specimens were fixed by perfusion and the permanent tooth buds were prepared for transmission electron microscopy. At the cervical loop proper, inner enamel epithelium cells have lobed nuclei, a paucity of cytoplasm, and wide extracellular spaces; the basal lamina facing the dental papilla is straight. With increasing distance from the tip of the cervical loop, the following changes occur gradually: (a) preameloblasts elongate from 15 to 45 µm, and their organelles, particularly mitochondria and profiles of rough endoplasmic reticulum, become more numerous; (b) extracellular spaces decrease between preameloblasts starting at the basal (infranuclear) end; (c) the basement membrane becomes convoluted and associated with aperiodic fibers; (d) preodontoblast projections penetrate the aperiodic fibers; (e) collagen fibers subjacent to the basement membrane increase in density, with particularly thick fibers paralleling the aperiodic fibers. These modifications occur within three-fourths of the distance from the tip of the cervical loop to the mineralization front. The condensation of preodontoblasts is followed immediately by predentin synthesis. Concomitantly, the basement membrane breaks down and the aperiodic fibers are engulfed by preameloblasts. Preameloblast projections penetrate junctional predentin, contact mineralized dentin, and enamel synthesis ensues. At this stage the ameloblast is 45 µm long, the nucleus is central or basal, the Golgi apparatus has migrated apically, but the Tomes' process has not yet formed. The results indicate that odontogenesis inMacaca monkeys more closely resembles human odontogenesis than does that in the murine rodents.
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  • 4
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    Calcified tissue international 25 (1978), S. 145-159 
    ISSN: 1432-0827
    Keywords: Bird egg shell ; Ultrastructure ; Calcification ; Electron diffraction ; Microanalysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The egg-shell of Japanese quail was studied by several techniques. Semithin sections (1μm thick) of non-decalcified shell were observed by normal and polarized light microscopy. Thin sections of non-decalcified shell, examined by transmission electron microscopy, permitted us to observe the forms and dimensions of crystals of calcite within different layers of the shell: mammilary layer, layer of cones, palissade layer and surface crystal layer. There appears to be two distinct zones in the layer of cones as well as in the superficial crystal layer. Electron microdiffraction revealed the orientation of calcite crystals in the columns. Some crystal defects (twins?) were described and the possibility of their artefactual formation during ultramicrotomy is discussed. Localization of Ca, Mg, P and S were made by X-ray microanalysis of semithin sections. This technique shows that shell membranes, and chiefly the true cuticle, are also mineralized but, in these layers, minerals are not crystallized. Otherwise the distribution of Mg is not uniform throughout the shell thickness; it is less concentrated in the external zone of the layer of cones. These results together with observation of developing shells by scanning electron microscopy allowed us to propose a scheme for shell organization of the quail egg. This organization was related with decalcification which occurs during hatching.
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  • 5
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    Calcified tissue international 27 (1979), S. 205-210 
    ISSN: 1432-0827
    Keywords: Intestine ; Calcium ; Binding ; Protein ; Uremia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary In order to assess the usefulness of intestinal biopsies as indicators of end-organ responsiveness to vitamin D in uremic patients, calcium binding activity and calcium binding protein (CaBP) content were measured in intestinal biopsies from 12 uremic patients (glomerular filtration rate〈5.0 ml/min) and 12 adult controls. Values for both were found to vary with the site of biopsy, highest values being obtained in the duodenal bulb, with lower values distally. Values for activity correlated with values for CaBP content in both normals and uremics and no difference was observed between groups. Levels of calcium binding activity and content of CaBP did not correlate with serum immunoreactive parathormone levels, but were directly related to circulating 25-hydroxycholecalciferol (25-OHD) levels. The data show that intestinal CaBP is normal in activity, quantity, and affinity for calcium in malabsorbing uremic patients, and are consistent with the hypothesis that calcium malabsorption in uremia is unrelated to deficiency of intestinal calcium binding protein.
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  • 6
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    Calcified tissue international 24 (1977), S. 173-177 
    ISSN: 1432-0827
    Keywords: Enamel ; Calcium ; Diffusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The diffusion of45Ca ions in sound bovine enamel at pH 7.0 was studied as a function of temperature in the range 20°–40°C. The experimental data show that the diffusion is governed by two processes with activation energies of about 0.7 and 1.3 eV respectively. These values are considerably larger than the activation energy of calcium ion diffusion in water (0.2 eV). The diffusion process with the activation energy of 1.3 eV is dependent upon the calcium concentration in solution. A model based on the complex pore structure of enamel and on interactions of diffusing ions with the enamel constituents is discussed.
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  • 7
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    Calcified tissue international 24 (1977), S. 215-222 
    ISSN: 1432-0827
    Keywords: Mineralization ; Osteodentin ; Intracellular ; Ultrastructure ; Microanalysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Newly formed osteodentin obtained from the anterior extremities of fetal or young rat incisors was observed by means of electron microscopy and electron probe X-ray microanalysis. Cells related to osteodentin formation frequently showed membrane bound intracellular bodies containing varying amounts of fine, needle-shaped crystals, which were identified as apatite. The intracellular clusters of apatite crystals were extruded from the cells through membrane fusion or cellular degeneration. These extracellular clusters seemed to be gradually incorporated into the mineralizing collagenous matrix, which developed around them. Frequent occurrence of dense, dotshaped or filamentous profiles suggested that the dense bodies seen in the perinuclear regions or in the Golgi area were the sites of crystal formation. Energy dispersive X-ray point analysis showed that the intracellular or extracellular apatite clusters contained sulfur in a concentration higher than was present in the mineralizing collagenous matrix. Furthermore, wave dispersive X-ray line analysis showed that the concentration of sulfur was higher in the osteodentin matrix than in the dentin matrix. The sulfur detected is presumed to be contained in acid mucopolysaccharides, which were distributed more heavily in the osteodentin matrix than in the dentin matrix. On the basis of these data, it was concluded that the unique chemical and structural characteristics of the osteodentin result primarily from the incorporation of apatite clusters of intracellular origin and associated acid mucopolysaccharides.
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  • 8
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    Calcified tissue international 28 (1979), S. 131-136 
    ISSN: 1432-0827
    Keywords: Calcium ; Phosphate ; Citrate ; Precipitation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The influence of citrate ions on the precipitation of crystalline apatitic precipitates with low Ca/P molar ratios [octacalcium phosphate (OCP) and calcium-deficient apatites (DA) (system A)] and of the intercrystalline mixtures of calcium hydrogen phosphate dihydrate (DCPD) and DA (system B) was investigated. Samples were prepared by direct mixing of calcium chloride solutions (A, 6·10−3 mol dm−3; B, 1·10−1 mol dm−3) and sodium phosphate solutions (A, 6·10−3 mol dm−3; B, 2·10−2 mol dm−3) containing citrate (0–2·10−3 mol dm−3) and preadjusted to pH 7.4. In the presence of citrate ions: (a) crystal growth of OCP and DA was slowed down; (b) habit modification of DCPD crystals occurred; and (c) equilibration in intercrystalline mixtures of DCPD and DA's was slowed down. All phenomena were caused by surface adsorption of negatively charged ions, most probably CaC6H5O7-, which is the prevalent calcium citrate species under the given experimental conditions. Habit modification of DCPD was induced by preferential adsorption at the (001) crystal plane.
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  • 9
    ISSN: 1432-0827
    Keywords: Bone resorption ; Osteogenesis ; Fish bone ; Osteocytes ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The comparative ultrastructure of fish bone osteogenesis and resorption induced by scale removal was described in the osteocytic (cellular-boned)Carassius auratus and the anosteocytic (acellular-boned)Tilapia macrocephala. Osteocytes, present in osteocytic bone, were lacking in anosteocytic bone. In osteocytic bone the osteoblast secreted a collagenous preosseous matrix in which it became enmeshed and then was termed a preosteocyte. When the preosseous matrix mineralized, the preosteocyte was termed an osteocyte and was completely surrounded by bone. In anosteocytic bone the osteoblasts receded from the mineralizing front and never became trapped as osteocytes. During resorption, types A and B resorptive cells, present in both bone types, invaded the matrix and demineralized the osseous zone. These cells were characterized by large amounts of granular endoplasmic reticulum and intracellular inclusions containing crystal-like material. Although functionally similar to mammalian osteoclasts, these cells lacked a characteristic ruffled border and were not multinucleated. The osteocytes of cellular bone did not appear to be involved during demineralization.
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  • 10
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    Calcified tissue international 30 (1980), S. 209-216 
    ISSN: 1432-0827
    Keywords: Calvarium ; cAMP ; Vitamin D3 metabolites ; Calcium ; Parathyroid hormone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Results from in vitro works suggest that 1,25- and 24,25-dihydroxyvitamin D3 (1,25-(OH)2D3 and 24,25-(OH)2D3) act on bone via different mechanisms. The present investigation was performed to study the effect of these two metabolites and of their precursor 25-hyxdroxyvitamin D3 (25-(OH)D3) on bone cAMP content in vitro. Rats' paired half calvaria were incubated under sterile conditions with one vitamin D3 derivative (10−13 to 10−9 M) or with ethanol (0.005 ml for 15 min to 24 h in 1 ml medium containing 0, 0.2, 1, 2, or 3 mM calcium. In some experiments: (a) cycloheximide (10−5M) was added simultaneously with the vitamin D3 metabolites; (b) 1–84 bPTH (5 × 10−8 M) was added for 5 or 15 min at the end of the 24 h incubation. Calvaria were immersed in 1 ml TCA 5% 4°C and homogenized. The cAMP was extracted with diethylether and measured by a competitive protein binding assay. Results bring further evidence for a particular effect of low doses of 24,25-(OH)2D3 (10−9 to 10−12M) and of 25-(OH)D3 (10−9 to 10−11M) on bone, different from that of 1,25-(OH)2D3: cAMP content was higher in 24,25-(OH)2D3- or 25-(OH)D3-treated and lower in 1,25-(OH)2D3-treated calvaria than in ethanol-treated ones with 1 mM calcium. The 1,25-(OH)2D3 effect persisted in calcium-free medium whereas 25-(OH)D3 and 24,25-(OH)2D3 effects could not be observed with 0 mM nor with 3 mM calcium. The required duration of the preincubation (over 1 h) as well as the inhibitory action of cycloheximide may suggest an involvement of protein synthesis in the vitamin D3 metabolites effects. Neither 1,25-(OH)2D3 nor 24,25-(OH)2D3 affected the PTH-induced increase in bone cAMP content.
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  • 11
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    Calcified tissue international 32 (1980), S. 201-206 
    ISSN: 1432-0827
    Keywords: Magnesium ; Parathyroid hormone ; Secretion ; In vitro ; Calcium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary In a well-defined in vitro perifusion system, the effects of extracellular magnesium concentration (Mg) on parathyroid hormone (PTH) secretion by bovine parathyroid tissue were examined. At Mg less than 0.8 mM, the ability of the glands to secrete hormone maximally in response to low calcium (Ca) stimulation was progressively impaired. Low Mg also impaired the ability of isoproterenol, dibutyryl cyclic AMP and theophylline to stimulate hormone release. The defect in hormone release at low Mg observed in vitro was analogous to the well-documented inhibition of secretion observed in vivo. Increases in Mg from 0 to 0.8 mM rapidly repaired the defect in hormone secretion. At Mg above 1.0 mM there was a Ca-like effect on hormone release, with a progressive decrease in secretion at increased Mg. Although its mechanism is not yet clear, the low Mg effect appears to impair principally the process of hormone release rather than its biosynthesis or storage.
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  • 12
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    Calcified tissue international 36 (1984), S. 139-144 
    ISSN: 1432-0827
    Keywords: 1,25(OH)2-vitamin D ; Weight loss ; Phosphate ; Calcium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary During a review of 42 metabolic studies in healthy women and men we observed that serum 1,25-(OH)2-D concentrations were directly correlated to the observed daily changes in body weight (r=0.68;P〈0.001) and to caloric intake/kg/day (r=0.39;P=0.01). These relationships could not be accounted for by related and physiologically expected changes in serum Ca or iPTH concentrations. However, serum 1,25-(OH)2-D concentrations were observed to be inversely correlated to serum PO4 levels (r=−0.44;P=0.004). In addition, serum PO4 levels were inversely correlated to the daily changes in body weight (r=−0.40;P=0.009). Since dietary sodium intake averaged 142 mmol/day, it is unlikely that the observed changes in weight were the result of changes in salt and water balance. Thus it seems reasonable to speculate that serum 1,25-(OH)2-D concentrations may vary directly with energy balance, as reflected by changes in body weight. This effect may be mediated by alterations in PO4 metabolism. The accurate assessment of serum 1,25-(OH)2-D levels thus appears to require several measurements over time periods during which body weight is stable.
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  • 13
    ISSN: 1432-0827
    Keywords: Electromagnetic field ; Bicarbonate ; Phosphate ; Calcium ; Fluoride ; Osteogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The present studies are aimed at establishing molecular correlations in the interaction of very low frequency electromagnetic fields with biological systems. Ca-uptake by chick embryo tibia rudiment in short-term culture was a useful model. Tibiae of 8- to 10-day-old chick embryos were incubated 60 min in simplified culture media in the presence of45Ca at 37.5±0.5°C either inside or outside pulsating electromagnetic fields. Radioactivity count in the medium was the most accurate method for determining Ca-uptake by the rudiment. The effect of the fields on the Ca-uptake depended markedly on the chemical composition of the culture medium: bicarbonate was indispensable; glucose or sucrose was important; phosphate was potentiating; ethanol, Mg2+, and NaF were stimulating. The field had no effect in (a) blank medium without tibia, (b) tibiae that had been altered by fixation with aqueous glutaraldehyde, (c) nonliving artificial systems endowed with great or small ion sorption capacity. The unique bicarbonate effect with living systems and the passive behavior of nonliving ion sorbing systems prompt the suggestion that the electromagnetic field probably couples with specific processes, such as a bicarbonate-dependent Ca2+ ATPase and the active ion transport, at the cell membrane level. The molecular mechanisms remain to be established.
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  • 14
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    Calcified tissue international 36 (1984), S. 194-199 
    ISSN: 1432-0827
    Keywords: Benzo(B)Thiophene-2-Carboxylic Acid ; Bone cells ; Calcium ; Cyclic AMP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The purpose of the present study was to investigate the mechanism of action on bone of Benzo(B)Thiophene-2-Carboxylic Acid (BL-5583). BL-5583, at a dose range of 0.01–100 µg/ml, inhibited spontaneous as well as A23187 and PTH-induced bone resorption in tissue culture. This compound also decreased calcium uptake in both osteoclastic and osteoblastic enriched bone cell populations obtained by sequential collagenase digestion of 1–2 day newborn rat calvariae. The decrease occurred after a 5 min. incubation with45Ca and BL-5583. The effective dose range was 0.01–100 µg/ml. No effect on leucine incorporation or lactic acid production by bone cells was observed. BL-5583 also induced a transient decrease in calcium uptake in skin cells isolated from fetal rats by collagenase digestion, suggesting a lack of tissue specificity for this compound. No effect on cyclic AMP in isolated bone cells was observed with the same dose range that produced a calcium effect.
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  • 15
    ISSN: 1432-0827
    Keywords: Saliva ; Calcium ; Phosphate ; Ion-binding ; Supersaturation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Previous ultrafiltration studies indicated that up to one-half of the calcium and two-thirds of the phosphate in human salivary secretions may be bound by salivary proteins. Since this binding is an important variable in determining the extent of salivary supersaturation with respect to calcium phosphate salts, and since the amount of binding reported is surprisingly large, calcium and phosphate ion-binding by salivary macromolecules has been reexamined. From experiments using equilibrium dialysis, it was found that (1) the fraction of salivary calcium involved in macromolecular complexes ranges from a few percent for unstimulated secretions, to no more than about 10% for stimulated glandular salivas, and (2) salivary proteins do not bind phosphate ions to any significant extent. These findings, and experiments using an improved ultrafiltration membrane, indicate that the earlier results were artifacts of the ultrafiltration technique. Fractionation of salivary proteins, followed by equilibrium dialysis measurements, showed that the anionic proline-rich proteins and a basic proline-rich glycoprotein are responsible for most of the calcium binding now observed. The finding that macromolecular complexes of salivary calcium and phosphate have been overestimated in the past, leads to the conclusion that salivary calcium and phosphate ion activities in stimulated salivary secretions may be up to 50 to 100% higher than previously thought. Revised values were therefore used to recalculate the degree of salivary supersaturation with respect to calcium phosphate salts. The results indicate that stimulated salivary secretions are supersaturated with respect to dicalcium phosphate dihydrate; this is a substantially greater degree of supersaturation than previously reported.
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  • 16
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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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  • 17
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    Mycopathologia 59 (1976), S. 117-123 
    ISSN: 1573-0832
    Keywords: Ultrastructure ; Zygospore ; Mycorrhizal fungus ; Flaming crown
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The ultrastructural organization of the spores of the sporocarp of Endogone flammicorona was studied. Two types of organization are described. Initially the spore possessed a vacuolate protoplasm and was bound by two cell wall layers. The spore was surrounded by a hyphal mantle formed of a sheet of vacuolized hyphae with uniformly thin walls. Secondly, although the ultrastructural features of the spore appeared the same, it was now surrounded by a hyphal mantle with unevenly thickened walls (i. e., the so-called flaming crown) due to the gradual and irregular deposition of granules and lamellae. This crown gives the spore its most commonly observed morphological feature and is the preminent character employed taxonomically to speciate Endogone flammicorona Trappe & Gerdemann.
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  • 18
    ISSN: 1432-041X
    Keywords: Ultrastructure ; Scanning cytophotometry ; Chromatin ; Chondrocytes ; Regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Description / Table of Contents: Résumé Les cellules cartilagineuses des membres postérieurs deTriturus cristatus en régénération après amputation, ont été étudiées en microscopie électronique et par cytophotométrie à balayage. Nous nous sommes intéressés à la structure et à la distribution de la chromatine mais aussi à différents organites cytoplasmiques. Dans l'étude de cytophotométrie à balayage, la chromatine a été considérée à travers son constituant majeur, l'ADN, coloré par la réaction de Feulgen. Au cours de la régénération du membre, l'hétérochromatine initialement condensée, essentiellement accolée à la membrane nucléaire se décondense. Les vacuoles du cytoplasme, caractéristiques des animaux âgés par rapport aux animaux jeunes, disparaissent, les mitochondries et le reticulum endoplasmique rugueux deviennent plus abondants. Les caractéristiques nucléaires de l'activation cellulaire apparaissent précocement, précédent les modifications cytoplasmiques et conduisent à des cellules en tous points identiques aux cellules d'animaux jeunes en dehors de tout processus régénératif. Cette phase d'euchromatisation et de restructuration cytoplasmique est peut-être nécessaire à l'accroissement d'activité métabolique et à la division cellulaire qui suivent. Son déroulement peut expliquer tout au moins le ralentissement de la régénération observé chez les animaux âgés par rapport aux animaux jeunes.
    Notes: Summary Cartilaginous cells of aged newts (Triturus cristatus) were studied during hind limb regeneration. The electron microscope was used to study the structure and distribution of chromatin in the cell nuclei, while the DNA content of the chromatin was measured by means of a scanning cytophotometer. Changes in the ultrastructure of the cytoplasm during regeneration were also studied. It was observed that the structure and distribution of chromatin in the activated cell is greatly modified. In the non-activated cell of the aged newt, the chromatin is found highly condensed and distributed peripherally close to the nuclear membrane. In contrast, in the activated cells, the chromatin is much less condensed and is distributed throughout the nucleus. Moreover, cytoplasmic vacuoles, found only in the non-activated aged cells, disappear and an increase in the mitochondria and rough endoplasmic reticulum is also observed. Changes in the nuclear structure are observed prior to the cytoplasmic modifications. It is interesting to note that the process of activation induces structural changes in the aged cells which make these cells appear to be structurally identical to the young cells. This process of rejuvenation takes 3–5 days in the newt. We suggest that these structural changes of the chromatin and cytoplasm in the aged cells are necessary to increase the metabolic activity which precedes cell division. It may also explain why regeneration takes a longer time in the aged animals than in the young ones.
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  • 19
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    Development genes and evolution 192 (1983), S. 42-44 
    ISSN: 1432-041X
    Keywords: Chick embryo ; Gastrulation ; Adenylate cyclase ; cAMP phosphodiesterase ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The ultrastructural localization of adenylate cyclase (E.C. 4.6.1.1.) and cAMP phosphodiesterase (PDE) (E.C. 3.1.4.17.) in the ectoderm of the developmental stage 4 chick embryo was studied. Adenylate cyclase was localized in the lateral surfaces of the ectodermal cells. In the primitive streak cells the enzymatic activity was observed on all the lateral surfaces, whereas in the periphery of the blastoderm the reaction product was localized in the apical parts of the lateral plasma membranes only. cAMP PDE localized in the apical cytoplasm of the ectodermal cells, with highest activity in the globular projections.
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  • 20
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    Development genes and evolution 183 (1977), S. 233-248 
    ISSN: 1432-041X
    Keywords: Cytoplasmic architecture ; Ultrastructure ; Insect egg ; Pattern formation ; Yolk ; Cytoplasma-Architektur ; Ultrastruktur ; Insekten-Ei ; Musterbildung ; Dotter
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Description / Table of Contents: Zusammenfassung 1. Das Ei der ZuckmückeSmittia spec. wurde licht- und elektronenmikroskopisch untersucht. Die vorliegende Arbeit beschreibt den Bau des Periplasmas und des Dotter-Endoplasma-Systems vor Bildung der Polzellen. 2. Das Periplasma, nach außen vom Oolemm und einer mehrschichtigen Eihülle begrenzt, besteht aus einer ribosomenreichen cytoplasmatischen Matrix, in die vor allem Mitochondrien und ER-Zisternen, wenig annulate lamellae und gelegentlich Golgi-Apparate eingelagert sind. Mikrotubuli wurden nur selten nachgewiesen. Öfters sind Anhäufungen einer dichten granulierten Substanz zu beobachten, die in ihrer Struktur dem Oosom-Material ähnelt. 3. Das Dotter-Endoplasma-System stellt ein Netzwerk aus Cytoplasma dar, in das Proteid-Dotterkugeln, Lipidtröpfchen sowie Glycogen-Anhäufungen eingelagert sind. Das Endoplasma, das sich zu 3–7 Plasma-Inseln erweitern kann und unmittelbar in das Periplasma übergeht, besteht wie dieses aus einer cytoplasmatischen Matrix und enthält die gleichen Zellelemente wie das Periplasma. Rosettenförmige Membran-Strukturen werden als “nuclear envelope organizing center” gedeutet. 4. Drei der sorgfältig analysierten Eier enthielten je 2 Kerne; sie lagen in Plasma-Inseln in der hinteren Eihälfte. 5. Sowohl im Periplasma wie im Dotter-Endoplasma-System sind alle Zellelemente unregelmäßig verteilt. Eine besondere Anordnung oder Zonierung ist nicht zu erkennen. 6. Die räumliche Verteilung der erfaßten Eikomponenten liefert keine Hinweise auf eine Funktion dieser Komponenten als Determinanten für die embryonale Musterbildung.
    Notes: Summary 1. Eggs of the midgeSmittia were investigated by light microscopy and transmission electron microscopy. This paper describes elements and architecture of periplasm and yolk endoplasm before the formation of pole cells. 2. The periplasm is coated externally by the oolemma and a multilayered egg shell. The periplasm consists of a cytoplasmic matrix rich in ribosomes; it contains mitochondria and ER cisternae, some annulate lamellae and an occasional Golgi complex. Microtubuli were demonstrated only rarely. Accumulations of a dense granulated substance resembling in its structure the oosome material were frequently observed. 3. The yolk endoplasm is a cytoplasmic network embodying proteid yolk particles, lipid droplets and accumulations of glycogen. The endoplasm is continuous with the periplasm and shows the same cell constituents. It may form between 3 and 7 cytoplasmic islands free of yolk particles. Rosette-shaped membranous structures in the yolk endoplasm are interpreted as nuclear envelope organizing centres. 4. Three carefully analysed eggs contained 2 nuclei each. both nuclei were situated in the posterior egg half. 5. Periplasm and yolk endoplasm are characterized by random distribution of cell elements. No zonation or special accumulations could be recognized. 6. The spatial distribution of the egg components studied did not indicate that any of these components could function as a determinant in embryonic pattern formation.
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    Development genes and evolution 181 (1977), S. 333-355 
    ISSN: 1432-041X
    Keywords: Barnacle eggs ; Constriction rings ; Microfilaments ; Ultrastructure ; Peristalsis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary 1. The egg ofPollicipes polymerus, the common intertidal gooseneck barnacle, has been studied by electron microscopy. Constriction rings, similar to the contractile rings of cleaving cells and polar lobes, move unidirectionally from the animal to the vegetal pole of newly fertilized eggs. This is referred to as peristaltic constriction. The present paper describes the fine structure of the egg during first polar body formation and peristalsis. 2. During formation of the polar body, dense bodies are produced by the Golgi and extracellular plaques are observed. Thin microfilaments (40–60 Å) are in the egg adjacent to the polar body. 3. In eggs undergoing peristalsis, the appearance of extracellular spheres, flocculent material and filaments is observed. Intracellularly large numbers of multivesiculate bodies, glycogen granules, mitochondria and protein-carbohydrate and lipid yolk bodies are seen at the level of constriction. 4. Thin microfilaments are found in the cortical area of newly-fertilized eggs exclusively in peristaltic constriction rings. Filaments are oriented primarily in a meshwork, although circumferentially-oriented filaments are also found in rings near the vegetal pole. Microvilli extend into the space created between a constriction and the elevated egg membrane. 5. A model is proposed to explain the peristalsis in this species. It is suggested that information from a pacemaker region activates peristalsis by affecting filament polymerization and orientation. One function of peristalsis may be elongation of the egg from a sphere to an ovoid, although other possibilities such as elevation of the egg membrane, segregation of the lipid yolk to the vegetal pole and predetermination of the first cleavage plane are also discussed.
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    Development genes and evolution 186 (1979), S. 65-70 
    ISSN: 1432-041X
    Keywords: Sea urchin ; Embryo ; Collagen ; Ultrastructure
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    Topics: Biology
    Notes: Summary Collagen fibrils with a main period banding of 610 Å and 220 Å in width were observed in the blastocoel of 72-h embryos of the sea urchin,Strongylocentrotus purpuratus. Non-striated fibrils of 50 Å diameter were also observed. The collagen is seen in highest concentration in the vicinity of mesenchyme cells which are richly endowed with endoplasmic reticulum and secretory vesicles. A role for collagen in cell attachment, orientation and spicule formation is discussed.
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    Development genes and evolution 192 (1983), S. 171-178 
    ISSN: 1432-041X
    Keywords: Differentiation ; Digestive tract ; Endoderm ; Organ culture ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The self-differentiation potency of the endoderm of the chick embryo was investigated mainly by transmission electron microscopy. Endodermal fragments isolated from 4- to 6-day stomach or small intestine were cultured in the absence of mesenchyme and were able to differentiate in vitro into organ-specific epithelia. Endodermal fragments isolated from the stomach region differentiated into a pseudo-stratified epithelium with periodic acid Schiff-positive mucous granules in the apical cytoplasm, while those from the small intestinal region differentiated into a simple columnar epithelium with a striated border which was positive in alkaline phosphatase activity. These features are comparable with those of the mucous secretory epithelium of the normal embryonic stomach and the absorptive epithelium of normal embryonic small intestine, respectively. Next, the self-differentiation potencies were investigated of the upper and lower layers of the blastoderms, at stages 1–5 of Hamburger and Hamilton (H. and H.). Both stomach-type and small-intestine-type epithelia developed only when fragments of the lower layer isolated from the blastoderms older than stage 3 of H. and H. were cultured, suggesting that cells possessing the potency to differentiate into the stomach- and small-intestine-type epithelia exist in the definitive endoderm at the beginning of its formation.
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    Development genes and evolution 188 (1980), S. 65-73 
    ISSN: 1432-041X
    Keywords: Nuclear migration ; Cleavage ; Microtubules ; Ultrastructure ; Gall midge
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    Topics: Biology
    Notes: Summary In the eggs ofWachtliella persicariae the cleavage nuclei move relative to the surrounding ooplasm. This ‘active’ migration is caused by an organelle whose ultrastructure was studied throughout the mitotic cycle. It consists of a greatly enlarged polar cytaster derived from the mitotic apparatus, linked to the nucleus by 100 Å filaments. The microtubules of the cytaster were found only during periods of active nuclear migration, i.e., from the onset of anaphase to the early prophase of the next mitotic cycle. They are always solitary and follow the course of the astral rays, which are known to temporarily adhere to peripheral structures of the egg cell and to exert tractive forces. In contrast to the cytaster microtubules, the microtubules in the spindle are bundled and persist from early metaphase through late telophase. During ontogenesis the first migration cytaster is built up between 3 and 12 min after oviposition near the anterior egg pole, in the vicinity of the sperm nucleus. In non-inseminated eggs time lapse films show a migration cytaster to develop autonomously in a region free from nuclei, but it does not follow the normal path of the male pronucleus. In several cases the female pronucleus, which remains without a cytaster of its own, was observed to move to the cytaster generated in the absence of the male pronucleus. Whether or not it is adhering to a nucleus, the cytaster divides into two at the correct time, i.e, corresponding to the first cleavage division in fertilized eggs. In some non-inseminated eggs this type of ‘pseudocleavage’ has been observed to occur repeatedly, giving rise to an increasing number of anucleate cytasters.
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    Development genes and evolution 188 (1980), S. 163-177 
    ISSN: 1432-041X
    Keywords: Yolk sac ; Ultrastructure ; Embryogenesis ; Drosophila
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    Topics: Biology
    Notes: Summary Changes at the ultrastructural level during germ band extension in the embryo ofDrosophila melanogaster are described. Cytoplasmic connections between cells and the yolk sac are present during initial cellular movements. At this time, a continuous system of microfilaments is present adjacent to the membranes in the connections and at the periphery of the yolk sac. As germ band extension progresses, this system becomes discontinuous, and microfilaments are apparent only in the immediate vicinity of the connections. Cytoplasmic connections are disassembled at approximately the midpoint of extension; at the same time, extensive membrane associations develop between germ band cells and between these cells and adjacent yolk sac membranes. Positioning and orientation of cytoplasmic connections suggest that the yolk sac, via these connections, is actively involved in the cellular movements of early germ band extension.
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    Development genes and evolution 185 (1978), S. 235-248 
    ISSN: 1432-041X
    Keywords: Liver ; Primary culture ; Ultrastructure ; Albumin synthesis ; Xenopus
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    Topics: Biology
    Notes: Summary Electron microscopic analysis of primary cultures derived from larvalXenopus liver has shown that these cells, although they form only two-dimensional aggregates, retain and presumably also develop structural characteristics typical of liver parenchyma cells, such as bile canaliculi with microvilli and epithelial junctional complexes. As judged from structural criteria, primary cultures contain 80–90% hepatocytes. In contrast to the intact tissue, primary cultures showed excessive development of microfilaments, however. Incorporation of labeled amino acids has revealed further that the capacity for protein synthesis is maintained in culture and that synthesis of liverspecific protein albumin is maintained in vitro, even in liver cultures derived from thyrostatic tadpoles. This latter result suggests that initiation of albumin synthesis in the larval liver is probably not dependent upon thyroid hormones but rather reflects the protodifferentiated state of this tissue.
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    Development genes and evolution 185 (1979), S. 333-346 
    ISSN: 1432-041X
    Keywords: Chick embryo ; Limb bud ; Ultrastructure ; Cell death
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    Notes: Summary The ultrastructural changes in the wing bud afterapical ectodermal ridge (A.E.R.) removal was studied to re-examine the issue of distal mesenchymal cell death. The A.E.R. of the right wing bud was removed microsurgically from chick embryos of stages 18 to 22 (HH 1951). The wing buds were examined at three hour intervals up to twelve hours after the operation with light, transmission and scanning electron microscopy. The main findings were: (1) Immediate and temporary shrinkage of the mesenchymal extracellular space 100 to 150 μm and chromatin condensation in the cells 50 to 75 μm from the wound. (2) Death of ectodermal and mesenchymal cells in the immediate vicinity of the wound. (3) Formation of a single squamous-like layer of mesenchymal cells to cover the wound. (4) Occasional evidence of cell death in the distal mesenchyme at later times after the operation. The pattern of cell death observed suggests only a traumatic etiology, and gives little evidence for the postulated developmental significance of cell death following A.E.R. removal.
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    Calcified tissue international 24 (1977), S. 59-64 
    ISSN: 1432-0827
    Keywords: Enamel ; Calcium ; Exchange ; Diffusion
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    Topics: Biology , Medicine , Physics
    Notes: Summary The penetration of45Ca ions into whole bovine enamel from an aqueous solution was studied as a function of the Ca content in the solution. A sectioning technique was employed to measure the penetration of45Ca into the enamel. Probably only Ca ions occupying surface positions in the hydroxyapatite crystallites participate in the exchange process at the solution-enamel interface. The fraction of Ca ions participating in the exchange was 2% of the total number of Ca ions. Two different diffusion processes occur; firstly, a process governed by a diffusion coefficient, D1 ranging from about 5×10−14 to 2×10−12 cm2/s and strongly dependent on the calcium concentration in the solutions; and secondly a process with a concentration-independent diffusion coefficient, D2, with a value of 2.8×10−12 cm2/s. It is assumed that the 2 diffusion types correspond to intra-and interprismatic transport, respectively. A model based on the pore structure of dental enamel and on surface charges on hydroxyapatite crystallites is discussed.
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    Calcified tissue international 24 (1977), S. 113-117 
    ISSN: 1432-0827
    Keywords: Bone membrane ; Calcium ; Ion fluxes ; Calvarium
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    Topics: Biology , Medicine , Physics
    Notes: Summary To facilitate the study of membrane function in the control of the flow of ions into and out of bone, it was desirable to develop a system for the direct quantitation of unidirectional effluxes of calcium and phosphate from bones. Based on a mathematical analysis of the problem, a specially designed Ussing chamber was developed which proved successful. Calvaria from 2-day-old rat pups, 3-day chicks and adult mice were evaluated. Calcium influxes which exceeded the corresponding effluxes were observed in the neonatal calvaria but not with those from adult mice. Also, an asymmetry in efflux was observed in rat calvaria, the inner side of the skull showed a higher efflux than did the external side. no such asymmetry was seen with calvaria from chicks or mice. This new technology should permit a further exploration of the role of the bone membrane in electrolyte homeostasis.
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    Calcified tissue international 22 (1977), S. 9-17 
    ISSN: 1432-0827
    Keywords: Bone ; Fluoride ; Calcium ; Phosphorus ; Mechanical Strength
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    Topics: Biology , Medicine , Physics
    Notes: Abstract Four groups of weanling rats were fed for 2 weeks on a diet sufficient or insufficient in calcium and/or phosphorus. Each group was divided into four subgroups which were offered distilled water supplemented with 0, 50, 75, or 150 ppm fluoride. High levels of fluoride in drinking water inhibited weight gain. This inhibition was less in rats deficient in phosphorus than when normal-phosphorus diets were offered. At a low level, fluoride was without any effect on bone ash, thickness of femoral cortical bone, and mechanical strength, as measured by maximal load, ultimate stress to breaking, and limit of elasticity. Modulus of elasticity was decreased. At higher levels fluoride tended to decrease most of these parameters, except in rats deprived of both calcium and phosphorus. The effect of fluoride was modified by lack of dietary calcium and/or phosphorus and appeared to be weaker in rats deficient in these nutrients. Lack of dietary calcium and/or phosphorus decreased bone strength more than did fluoride content of water and of bone mineral. Concentration of bone ash and thickness of femoral cortical bone were closely correlated with parameters of mechanical strength.
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    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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  • 32
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    Keywords: Calcium ; Intestine ; Growth ; Cortisol ; Hydroxyproline ; Parathyroid hormone
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    Topics: Biology , Medicine , Physics
    Notes: Summary This paper reports the effects of cortisol on intestinal management of calcium and on related changes on bone metabolism. Five groups of 12 rats each fed a standard diet (0.8% Ca) received 2, 6, 16, 32, and 128 mg/kg/day of cortisol hemisuccinate, subcutaneously. After 16 days, intestinal absorption and excretion of Ca were measured with the aid of45Ca. True Ca absorption increased as a function of dose up to 16 mg/kg/day and remained high with the larger doses. Endogenous fecal Ca excretion increased exponentially as a function of the dose from 16 mg/kg/day onwards. Therefore, a dual effect was observed: (a) an increase in true Ca absorption at low cortisol doses (which increased net Ca absorption); and (b) an increase in endogenous fecal Ca excretion at high doses (which reduced net Ca absorption). In no case was a depression of true Ca absorption observed. Growth rate and food conversion efficiency were depressed only with a cortisol dose of 128 mg/kg/day. The urinary excretion of hydroxyproline, pyrophosphate, and aminopolysaccharides decreased with low doses and increased above normal levels with the highest dose. When animals treated with 128 mg/kg/day of cortisol were fed Ca-enriched diets, net Ca absorption improved. Simultaneously, growth rate and food conversion efficiency approached normal values. In these experiments, net absorption of Ca was found to be inversely related to urinary excretion of hydroxyproline. The urinary rate of excretion of hydroxyproline is suggested as an indicator of the effect of a Ca supplement on cortisol affected connective tissue turnover.
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  • 33
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    Keywords: Shell formation ; Free nerve endings ; Ultrastructure ; Lymnaea stagnalis ; Biomphalaria pfeifferi
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    Topics: Biology , Medicine , Physics
    Notes: Summary The mantle edge of the freshwater pulmonate snailsLymnaea stagnalis andBiomphalaria pfeifferi was investigated with histochemical and ultrastructural methods. The mantle edge gland, which is involved in shell formation, consists of the periostracal groove and the belt. This belt appears to be composed of various regions. In the area of the periostracal groove a number of subepithelial gland cell types occur; these release their products into the groove. Between the groove cells ciliated free nerve endings terminate; the corresponding perikarya occur in the subepidermal connective tissue. Also in the posterior belt region free nerve endings were observed between the epithelial cells; in addition, a particular type of subepithelial gland cell was found in this area. The epithelial cells of this part of the belt have the ultrastructural characteristics of ion and water transporting cells; they are probably involved in calcium deposition and resorption. The possible role of the free nerve endings and of the subepithelial gland cells is discussed.
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    Calcified tissue international 26 (1978), S. 199-202 
    ISSN: 1432-0827
    Keywords: Fluoride ; Calcium ; Phosphate ; Parathyroid hormone
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    Notes: Summary Fluoride 27 mg was given perorally to 14 subjects. It was found that the serum fluoride concentration increased followed by a decrease to normal levels during 24 h. Serum concentrations of calcium and phosphate decreased initially whereas those of serum immunoreactive parathyroid hormone increased. After 24 h the measured quantities were within normal limits. The results are discussed with reference to mineralization of hard tissues and to fluoride treatment of osteoporotic patients.
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  • 35
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    Keywords: Solanum malacoxylon ; Vitamin D metabolites ; Rickets ; Phosphate ; Calcium
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    Notes: Summary The effects of a partially purified aqueous extract ofSolanum malacoxylon (SM) leaves and of an SM extract incubated with ruminal fluid (RF) prior to administration on intestinal phosphate and calcium absorption in rachitic rats were compared with 1α, 25-dihydroxy-vitamin D3[1α,25-(OH)2-D3] using a duodenal loop in situ method. The time course responses elicited by both extracts were qualitatively similar to that of 1α, 25-(OH)2-D3. However, a single dose of SMRF was significantly more effective in stimulating32P absorption than an equivalent dose of SM. Part of this difference can be accounted for by incomplete saturation of the phosphate transport system with the level of SM employed. Treatment with rumen potentiated the effects of SM on45Ca absorption to a lesser extent. Extraction of SMRF with nonpolar organic solvents and purification of the lipid extract by chromatography on Silica Gel G thin-layer plates and on Sephadex LH-20 columns yield two fractions which promote intestinal32P absorption. The bioactive fraction eluting first from the Sephadex LH-20 column migrates similarly as 1α,25-(OH)2-D3. The greater effectiveness of SMRF extracts on intestinal phosphate and calcium absorption may be tentatively attributed to the release of 1α,25-(OH)2-D3 from the corresponding glycoside, shown previously to be present in SM, and to steroid modification during incubation with rumen.
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    Calcified tissue international 28 (1979), S. 17-22 
    ISSN: 1432-0827
    Keywords: Anticonvulsant ; Ketogenic diet ; Calcium ; Vitamin D ; Bone
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    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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    Calcified tissue international 27 (1979), S. 171-176 
    ISSN: 1432-0827
    Keywords: Calcium ; Phosphate ; Precipitation ; Double diffusion ; Collagen
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    Notes: Summary One-dimensional double diffusion was applied to determine critical concentrations at which the precipitation of calcium phosphates occurs in reconstituted connective tissue collagen and agar gels at 37°C and in gelatin gels at 25°C. Experiments were performed in the presence of unbuffered 0.15 mol dm−3 NaCl, or 0.15 mol dm−3 NaCl-veronal adjusted to pH 7.4. It was found that critical concentrations of precipitation of both precipitating components, CaCl2 and phosphate buffer (pH 7.4), were equimolar and independent of the ratios of initial concentrations of the components. Critical concentrations of precipitation were not affected by the concentrations and kinds of gels used. The first-formed precipitates showed amorphous structure by X-ray diffraction analyses. Infrared (IR) spectra of the precipitates indicated CaHPO4 · H2O to be their predominant species. The molar Ca/P ratio obtained by chemical analyses was 1.08. This precipitate transformed in time into octacalcium phosphate. In all experiments, two very thin membranes of precipitate were formed in the gel column at the onset of precipitation simultaneously on both sides of the actual disc of precipitate. IR spectra and chemical analyses showed that both membranes were identical to the actual precipitation discs.
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    Calcified tissue international 25 (1978), S. 227-232 
    ISSN: 1432-0827
    Keywords: Parathyroid hormone ; Bone ; Calcium ; Chick embryo ; Organ culture
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    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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    Calcified tissue international 32 (1980), S. 229-236 
    ISSN: 1432-0827
    Keywords: Bone ; Ion influxes ; Calcium ; Phosphate ; Exchange
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    Topics: Biology , Medicine , Physics
    Notes: Summary Quantitative measurements were made of the ion fluxes of calcium and phosphate into and from calvaria (mouse or rat) when clamped in specially designed micro-Ussing chambers. The effects of varying concentrations of calcium were examined on the influx and efflux of calcium and of its counterion, phosphate. A comparable series of experiments was performed with varying phosphate concentrations. Both ions, as their concentrations increased, depressed their own influx, increased their own efflux, and significantly increased the equilibrium concentration, E/K, supported by the calvaria. Similarly, both ions, as their concentrations increased, affected the influx or efflux of their counterion only slightly but did depress the counterion's equilibrium level, E/K, significantly. In spite of these changes it was shown that calvaria effectively buffered the medium at physiological concentrations of calcium and phosphate. The buffering capacity, however, was small, and the balance, E/K, was modified by small uptake or loss of either ion. The small size of the interacting mineral pool was confirmed by direct measurement of the rapidly exchanging fractions of both calcium or phosphate. They were only ∼1% of the total ions present. The significance of these findings is discussed.
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  • 40
    ISSN: 1432-0827
    Keywords: Calcium ; Vitamin D deficiency ; 1,25(OH)2D3 ; Parathyroidectomy ; Parathyroid hormone
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    Notes: Summary Studies presented here were designed to investigate further the basis for an impaired cAMP response to parathyroid hormone (PTH) in osteoblastlike calvarial bone cells isolated from vitamin D-deficient rat pups. The goal was to perturb Ca, PTH, and vitamin Din vivo in order to see which factors might be responsible for the impairedin vitro bone cell cAMP response. Pups either were parathyroidectomized (PTX) 3–5 days, implanted with osmotic minipumps delivering high doses of PTH, given repeated, high doses of 1,25(OH)2D3, or were D-deficient (-D, i.e., born and suckled by D-deficient mothers). Osteoblastlike bone cells, isolated by sequential enzyme digestion and centrifugation, were exposed to PTH for 5 min in the presence of a phosphodiesterase inhibitor. In bone cells isolated from -D rat pups, both basal and PTH-induced cAMP accumulation were significantly lower than in +D bone cells. Earlier, we had shown that two daily injections of -D pups with 50 ng 1,25(OH)2D3 restores this reduced bone cAMP response of -D pups toward normal. In the present study, neither basal nor PTH-induced bone cell cAMP accumulation was affected by subjecting D-replete pups to PTX, PTH infusion, or repeated high doses of 1,25(OH)2D3 despite the fact that each treatment markedly changed serum Ca or serum immunoreactive PTH. The results indicate that the impaired bone cell cAMP response seen in -D pups is not a direct result of chronic hypocalcemia and that the “heterologous desensitization” seenin vitro with added 1,25(OH)2D3 could not be duplicated byin vivo treatment of +D pups with supraphysiologic doses of 1,25(OH)2D3. Finally the lack of alteration in the bone cell cAMP response to PTHin vitro after chronic PTH infusionin vivo fails to support the notion that the impaired response in -D bone cells can be explained entirely by “homologous desensitization” induced by high circulating levels of PTH in the hypocalcemic, -D rat pup.
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  • 41
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    Calcified tissue international 34 (1982), S. 273-279 
    ISSN: 1432-0827
    Keywords: Odontogenesis ; Ultrastructure ; Alkaline phosphatase
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    Topics: Biology , Medicine , Physics
    Notes: Summary The ultrastructural localization and gradient of activity of alkaline phosphatase were studied with respect to cell differentiation, matrix synthesis, and matrix mineralization in the incisor and molar teeth of 4-day-old Sprague-Dawley rats. The animals were perfused intracardially at room temperature with 2.5% glutaraldehyde in 0.1M sodium cacodylate (pH 7.4) with 3–4% sucrose. The jaws were dissected, immersion-fixed for 24 h, and the incisor and molar tooth germs removed. These were demineralized in 10% EDTA in NaOH (pH 7.4) with 7% sucrose. After reactivation of the enzyme with 0.1M MgCl in Tris-maleate buffer (pH 7.4) at 4°C, the teeth were incubated for alkaline phosphatase in a medium consisting of 6 ml 3% sodiumβ-glycerophosphate, 4 ml 0.2M Tris-HCl buffer (pH 9.2), 3 ml 1.6% MgSO4, 12 ml 0.5% lead citrate (pH⋍12), and 2.1 g sucrose. The pH was adjusted to 9.2 with 0.2M HCl, the volume made up to 30 ml, and the solution centrifuged for 10 min at 5000 rpm. Control teeth were incubated in medium minus the substrate. Finally, the specimens were routinely post-fixed and embedded for sectioning and examination with a Philips 300 electron microscope. A gradient of alkaline phosphatase activity was mapped along the developing teeth in the cells of the stratum intermedium, the proximal borders of the ameloblasts, the early dentine matrix, the predentine-dentine border, matrix vesicles, and the plasma membranes of odontoblasts and subodontoblast cells. The gradient of alkaline phosphatase activity was evident in the forming tooth from the cervical loop to the crown apex and was related to the cellular events, matrix synthesis, and matrix mineralization occurring during odontogenesis.
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  • 42
    ISSN: 1432-0827
    Keywords: Calcitonin ; Parathyroid hormone ; Calcium ; Newborn ; Fracture
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    Topics: Biology , Medicine , Physics
    Notes: Summary Determinations of serum calcium (Ca), phosphorus (P), calcitonin (CT), and parathyroid hormone (PTH) were carried out in 36 full-term newborn infants with fracture of the clavicle (CF) and in 46 normal neonates (N). At the 6th hour of life the CF neonates demonstrated lower serum Ca and higher serum CT in comparison with normal infants. In the hours following, no significant differences between the two groups for the Ca levels were found, whereas serum CT remained significantly higher in the CF newborns at the 24th, 48th, and 72nd hour of life. Significant differences between normal and CF infants in the PTH serum levels were detected only at the 48th hour, when PTH was lower in the CF newborns. The results of this investigation indicate that the fracture of the clavicle is a significant and peculiar factor in stimulating CT secretion. Serum Ca level appeared to be controlled by CT rather than auto-regulating the secretion of the hormone.
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  • 43
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    Calcified tissue international 33 (1981), S. 365-368 
    ISSN: 1432-0827
    Keywords: Absorptiometry ; Osteoporosis ; Spinal bone ; Bone mineral content ; Neutron activation ; Calcium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Dual-photon absorptiometry using153Gd (44 and 100 keV) was used to measure the bone mineral content (BMC) of the trunk and of the total body (TBBM) in 7 volunteers with no overt bone disease. These values were compared to those obtained with partial-body neutron activation of calcium (trunk Ca). The trunk Ca seemed to represent best a 60 × 30 cm area; the correlation coefficient with the corresponding BMC in that area was 0.97 (SEE ⋍ 7%). Trunk Ca was also highly correlated with TBBM (r=0.96; SEE=8%) and with radius BMC (r=0.92; SEE=11%), but the correlations with the BMC of smaller subareas of the trunk were lower (r⋍0.9; SEE ∼ 12%). The BMC of the lumbar spine was only moderately correlated with trunk Ca, radius BMC and TBBM (r ∼ 0.82; SEE ∼ 18%), and only slightly more associated with trunk BMC (r ∼ 0.88; SEE ∼ 14%). The BMC of the combined lumbar-thoracic spine showed higher correlations with trunk Ca, radius BMC and TBBM (r ∼ 0.87; SEE ∼ 13%), and trunk BMC (r=0.93; SEE ∼ 10%). An accurate and sensitive measure of spinal status requires a direct measurement of that area.
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  • 44
    ISSN: 1432-0827
    Keywords: Bone ; Calcium ; Strontium ; Apposition ; Tracer analysis
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    Topics: Biology , Medicine , Physics
    Notes: Abstract 1. Evidence has accumulated that the rate of acceretion (A) of calcium to bone is the sum of two fluxes; apposition involving the laying down of new bone and augmentation which is the result of slow exchange of non-surface bone calcium with plasma calcium pools as the result of solid state diffusion. 2. A method has been devised for separating A into its two components. It requires the use of45Ca or, for clinical studies,85Sr as a calcium tracer. Studies which are initiated with a combined accretion rate-calcium balance study, are concluded with an estimate of the exponent of the power function which has been found to describe the whole body retention of tracer from the second month onward. 3. The impulse response function of the skeleton for the tracer is then calculated, making the assumption that in any uniform volume of bone, osteoclastic resorption is a first order process. Making in addition certain simplifying assumptions, which are shown to have a modest influence on the final results, a mean rate of bone resorption can be calculated using a development of the well known Stewart-Hamilton formula. The apposition rate is calculated as the sum of the resorption rate and the calcium balance. Augmentation and diminution, defined as equal and opposite exchange processes, are given by the difference between A and the apposition rate. 4. The results of our first thirteen studies in normal subjects and patients with metabolic bone disease are presented, together with analyses of some data from the literature. It is concluded that the development of an atraumatic method for measuring rates of bone formation and resorption in the whole body would be an important advance in the study of metabolic bone disease, and this work is presented so that critical comparisons may be initiated between this tracer method and independent histological methods for measuring these parameters.
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  • 45
    ISSN: 1432-0827
    Keywords: Bone cells ; Cyclic AMP ; Calcium ; Ascites fluid resorptive protein
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    Notes: Summary The effects of a bone resorptive protein isolated from human cancer ascites fluid on bone cell calcium and cyclic AMP were studied with fetal rat cells. The osteoclast-activating factor increased bone cell calcium uptake at 37°C and 4°C with no direct effects on calcium efflux. Concentrations of the resorptive factor that increased in vitro bone resorption and cell calcium uptake had no effect on cyclic AMP. The effects of the protein on calcium uptake were not specific for bone cells, and large increases were also observed in isolated fetal rat skin cells. These studies suggest that increases in the permeability of the cell membrane to calcium are involved in the mechanism of action of the ascites fluid resorptive protein.
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  • 46
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    Calcified tissue international 23 (1977), S. 87-94 
    ISSN: 1432-0827
    Keywords: Calcium ; Osteoporosis ; Lactation ; Chemistry
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    Topics: Biology , Medicine , Physics
    Notes: Summary The calcium homeostatic mechanism was challenged in adult female rats by feeding them a calcium-deficient diet containing oxalate, and by subjecting them to pregnancy and lactation. The regimen caused a substantial weight loss, especially in those animals which reared their young well. Severe hypocalcaemia was observed in the lactating rats. Serum-P was slightly elevated. The content of hydroxyproline in serum was considerably elevated, reflecting the degree of calcium deprivation. Serum proteins were least influenced. The calcium depriving regimen reduced the growth of long bones, but did not stop it. The ash content of the bones was considerably reduced, the degree of reduction depended on the degree of calcium deprivation. Ash as percentage of total bone organ was reduced, but not to the same extent as total ash. Analyses of different parts of femur showed that the proximal and distal parts had lost more bone mineral than the diaphyseal shaft. The ash content of cortical bone tissue from the femur was estimated by a volumetric method. No differences were observed between test groups and controls, indicating that no measurable amounts of bone mineral had been removed from the walls of the vascular canals or by osteocytic osteolysis. Planimetric determinations on cross sections from femora disclosed that a great amount of bone had been removed from the endosteal surface of the diaphysis, while the periosteal surface demonstrated reduced bone apposition.
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  • 47
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    Calcified tissue international 26 (1978), S. 39-45 
    ISSN: 1432-0827
    Keywords: Solanum malacoxylon ; Bone culture ; Resorption ; Calcium ; Osteoclasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The water-soluble active principle of the plantSolanum malacoxylon (SM) was partially purified by column chromatography, and its bone-resorbing activity was tested by using an organ culture system of 18-day fetal mouse fibulae prelabeled with45Ca. Three series of experiments were performed. In the first series, the aqueous extract of SM was submitted to Sephadex G-25 chromatography to separate the active substances. From six collected fractions only two exhibited reproducible bone-resorbing activity as indicated by an increased release of45Ca from bone rudiments. In the second series, the active fractions were combined and lyophilized to a powder which was dissolved in the culture media to obtain the concentrations of 5, 25, 50, and 100 μg/ml. The addition of this saltfree material resulted in a dose-related accentuation of45Ca release from fetal mouse bones into the medium. Histologically, the treated fibulae showed greater numbers of osteoclasts than the controls and a disappearance of bone trabeculae. In the third series, the active material was further purified on Sephadex LH-20 column. From four collected fractions, only one showed bone-resorbing activity in vitro. It is concluded that the purified active factor of SM exerts a direct effect on bone. It stimulates the mobilization of bone calcium in vitro by increasing the population of osteoclasts. The nature of the active principle remains unknown.
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  • 48
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    Calcified tissue international 26 (1978), S. 71-79 
    ISSN: 1432-0827
    Keywords: Absorption ; Calcium ; Mitochondria ; Intestinal epithelium ; Potassium-pyroantimonate
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    Topics: Biology , Medicine , Physics
    Notes: Summary Intracellular transport of calcium from the apical to the basal-lateral region of the intestinal epithelial cell was invetigated in duodenum from normal fed, fasted, and calcium-loaded rats. The process was followed with time using electron microscopy with potassium pyroantimonate to precipitate calcium. The observations made were subjected to morphometric analysis. The specificity of the method was demonstrated in the villus cell by resistance to micro-incineration and by absence of deposits following exposure to EGTA. Using this method calcium was seen in cells from calcium-fed rats at the microvillus border, in the Golgi zone, and within the internal compartments of the mitochondria. In cells from fasted rats calcium was not seen. Mitochondria were found largely at the apex of the cell and were free of detectable calcium. By 5 min, in the cells of fasted rats given a calcium load, the calcium had reached the Golgi apparatus and the inner mitochondrial compartment. After 15 min mitochondria were heavily loaded with calcium and had moved to the basal region of the cell. These observations suggest that mitochondria play an important role in absorption of calcium and appear to transport this ion from the apex to the basal region of the cell where entry into the capillaries takes place.
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  • 49
    ISSN: 1432-0827
    Keywords: Parathyroid hormone ; Calcium ; Plasma phosphate ; Bone phosphate ; Plasma32P ; Specific activity
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    Topics: Biology , Medicine , Physics
    Notes: Summary Plasma changes in calcium, phosphate, and their radionuclides were studied in thyroparathyroidectomized (TPTX) rats treated with parathyroid hormone (PTH) for 8 h, this treatment starting 10 h after injection of45Ca and32P. Prior to intravenous infusion or hourly injections of PTH (10 mU/g/h), rats were maintained in one of three ways: on an extended fast (24 h); on a partial fast (10 h); or provided with 10% glucose and 1% calcium lactate overnight as a substitution for solid food. The pattern of change for plasma calcium,45Ca, and45Ca specific activity (S.A.) produced by PTH was not affected by these dietary conditions. The changes in phosphate were as follows: During the experimental (8 h) period, the rate of loss of32P from plasma in control rats was proportional to the length of the fast. This suggests that32P was released into plasma during the experimental period proportional to the ready availability of soft tissue glucose. In rats on an extended fast, PTH was phosphaturic, hypophosphatemic, and increased the rate of loss of32P from plasma without affecting32P S.A. values. In rats fasted for only 10 h, PTH produced similar effects on plasma phosphate and plasma32P values, but also caused a significant fall in plasma32P S.A. After glucose and calcium lactate treatment, PTH-induced phosphaturia was temporarily lost and the marked hypophosphatemia was replaced with a slight hyperphosphatemia. Plasma32P values also rose slightly; therefore, no effect on32P S.A. was produced. It is concluded from these studies that as the result of the phosphaturia caused by PTH, the hypophosphatemia which is produced automatically changes the phosphate gradient between various body compartments, causing phosphate entry into plasma. The authors postulate that this phosphate entering plasma is withdrawn primarily from bone fluid and bone.
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  • 50
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    Calcified tissue international 24 (1977), S. 271-274 
    ISSN: 1432-0827
    Keywords: Enamel ; Mineralization ; Calcium
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    Topics: Biology , Medicine , Physics
    Notes: Summary The potassium pyroantimonate technique was used to study the cellular distribution of calcium during the early mineralization of enamel in rat molar tooth germs at the electron microscope level. Differing patterns of calcium distribution were observed in the ameloblast seemingly associated with the appearance of Tomes' process. In the early secretory ameloblast calcium pyroantimonate deposits were observed within the Golgi apparatus, within coated vesicles, within mitochondria and associated with the inner aspect of the cell membrane. However, with the development of Tomes' process the ameloblasts no longer demonstrated these discrete deposits of calcium pyroantimonate. Instead they showed a diffuse cytoplasmic staining pattern with no preference for any particular organelle.
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  • 51
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    Calcified tissue international 27 (1979), S. 13-18 
    ISSN: 1432-0827
    Keywords: Anticonvulsants ; Calcium ; Vitamin D ; Parathyroid hormone ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Parameters of mineral metabolism were examined in 6 patients with moderately severe anticonvulsant drug-induced osteomalacia. Compared to 15 matched controls, the patients exhibited significantly reduced serum calcium, inorganic phosphate, and 25-hydroxyvitamin D concentration, elevated serum alkaline phosphatase and immunoreactive parathyroid hormone (iPTH) concentration, reduced intestinal47Ca absorption, reduced urinary calcium and increased urinary hydroxyproline excretion, and reduced forearm bone mass. Intestinal absorption of vitamin D3 was normal. Following 4 months of treatment with vitamin D3 (4000 units/day), serum 25-OHD concentration was increased to 3 times mean normal values and all parameters except serum iPTH, urinary calcium excretion, and forearm bone mass were returned to levels not significantly different from normal. Serum iPTH concentration was reduced by 39% (P〈0.05); 24-h urinary calcium excretion rose by 98% (P〈0.001), and forearm bone mass increased by 5.6% (P〈0.05). It is concluded that moderate-dose vitamin D3 supplementation is effective in normalizing parameters of mineral metabolism in this disorder, despite evidence of resistance to the biologic effects of vitamin D.
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  • 52
    ISSN: 1432-0827
    Keywords: Calcium ; PTH ; 1,25(OH)2D ; Acid ; Bone
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    Topics: Biology , Medicine , Physics
    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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  • 53
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    Calcified tissue international 29 (1979), S. 63-69 
    ISSN: 1432-0827
    Keywords: Vaterite ; Calcium ; Eggshells ; Cuckoos
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    Topics: Biology , Medicine , Physics
    Notes: Summary The crystalline spherules occurring in patches on the outer surface of the eggshells ofGuira guira and the bloom on the entire outer surface of those ofCrotophagi ani were shown by X-ray diffraction analysis to be vaterite.
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  • 54
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    Calcified tissue international 29 (1979), S. 155-161 
    ISSN: 1432-0827
    Keywords: Periosteal cells ; Calcium ; PTH ; CT ; Metabolic inhibitors
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    Topics: Biology , Medicine , Physics
    Notes: Summary Calcium transport was studied in cells isolated from cultured periostea of 18-day-old chick embryos. Net calcium uptake was stimulated by iodoacetate (IAA) and inhibited by dinitrophenol (DNP). Calcium efflux from the intracellular compartment was inhibited by IAA. Changes in the extra-or intracellular sodium concentration had only minor effects on calcium transport. This indicates that calcium efflux from periosteal cells is probably directly dependent on ATP hydrolysis, whereas calcium-sodium exchange is of less importance. The cation ionophore A23187 stimulated calcium uptake during short incubations but was inhibitory in long incubations. The possible involvement of the mitochondria in this effect is discussed. Parathyroid hormone (PTH) and calcitonin (CT) stimulated the net uptake of calcium in relatively low doses (both 0.01 U/ml or higher). The effects of the hormones on net calcium uptake were not additive. Calcium efflux was not changed in the presence of PTH or CT.
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  • 55
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    Calcified tissue international 31 (1980), S. 93-108 
    ISSN: 1432-0827
    Keywords: Calculus ; Ultrastructure ; Apatite ; Transmission ; Scanning
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    Topics: Biology , Medicine , Physics
    Notes: Summary Using transmission and scanning electron microscopy, we have studied the ultrastructure of a number of urinary calculi, mainly composed of calcium phosphate. Three fundamental kinds of calcium phosphates were detected: nonstoichiometric carbonate apatite, nonhexagonal octacalcium phosphate, and calcium-magnesium whitlockite. The influence that the organic matter, substitutions in the phosphate lattice of CO3 and Mg, and apatitic stoichiometry have on the ultrastructure of the calcium phosphate calculi has been detailed. An originating apatitic unity named U2 is assumed to be the responsible for all the different structures of calcium apatites appearing in renal calculi. On the basis of our observations, a mechanism whereby apatites grow is postulated; magnesium functions as an inhibitor for the growing mechanism.
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  • 56
    ISSN: 1432-0827
    Keywords: Hydroxyapatite ; Dissolution ; Pyrophosphate, Diphosphonates ; Calcium ; Phosphate
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    Topics: Biology , Medicine , Physics
    Notes: Summary Pyrophosphate and diphosphonate ions have been said to diminish the dissolution of hydroxyapatite crystals, because they lower the equilibrium concentrations of calcium and phosphate ions in the bulk solution around hydroxyapatite crystals in a closed system. However, in a closed system these effects are not necessarily due to an effect on dissolution alone. In this paper we have used a continuous flow system to study the effects of pyrophosphate and two diphosphonates, ethane-1-hydroxy-1,1-diphosphonate and dichloromethane diphosphonate, on the dissolution of hydroxyapatite. All three compounds decreased markedly the rate of dissolution of hydroxyapatite as well as the exchangeable pools of calcium and phosphate ions around the cystals.
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  • 57
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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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    Calcified tissue international 32 (1980), S. 45-53 
    ISSN: 1432-0827
    Keywords: Dentin ; Periodicity ; Allometry ; Calcium ; Sulfur
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    Topics: Biology , Medicine , Physics
    Notes: Summary We have described differences in the aspects of biological rhythms for calcium and sulfur deposition on the labial and lingual sides of the growing rabbit incisor, where growth occurs along a spiral axis. The calcium oscillations appear to be smoother on the labial side than on the lingual side. The lingual side is characterized by high-frequency rhythms with high amplitudes which possess the greatest percent of the power (Fourier analysis). These observations also reflect a difference in behavior of the mean Ca concentration across the labial and lingual sides. Sulfur rhythms on the labial side have higher amplitudes than those on the lingual side, but systematic differences in distribution of power between high and low frequencies is not as pronounced as in the case of Ca. The differences in Ca rhythms reflect differences in the growth rates of incisors on either side of the spiral axis. The labial side grows slightly faster than the lingual side, and its odontoblasts secrete Ca along the spiral axis and toward the pulp cavity at the same time. Thus the resultant direction of growth is more nearly opposite the extension of the occlusal end on the labial side, and Ca is consequently deposited over a wider area relative to that on the lingual surfaces. On the lingual side, Ca is deposited within a more limited area, and growth must therefore be continuous at high frequencies. The distribution of Ca on both sides of the tooth reflects these differences in growth rate and periodicity in two ways. First, given a unit area of tooth, the calcium concentration on the labial side is less than that of the lingual side. Second, whereas the calcium concentration on the labial side declines rapidly from the enamel-dentin junction to the pulp cavity, it is uniformly high across the lingual side because its growth is more continuous at high frequencies.
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  • 59
    ISSN: 1432-0827
    Keywords: 1α-Hydroxy vitamin D3 ; 1,25-Dihydroxyvitamin D3 ; Calcium ; Transport ; Intestine
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    Topics: Biology , Medicine , Physics
    Notes: Summary The timed sequence of events following the oral administration of 1α-hydroxy vitamin D3 (1αOHD3) to rachitic chicks was compared to that following a comparable dose of 1α,25-dihydroxyvitamin D3 (1,25(OH)2D3). RNA polymerase activity was maximally increased 20% by 1αOHD3 within 1 to 2 h and returned to control values after 8 h. Alkaline phosphatase activity was stimulated by 4 h and was maximal (3- to 5-fold increase) at 24 h. Calcium binding protein (CaBP) was detected initially within epithelial cells at the proximal end of the villus (just above the crypt) 6 to 8 h after 1αOHD3 administration, in epithelial cells lining the proximal half of the villus by 24 h, and in epithelial cells along nearly the entire villus by 48 h. At no time did goblet cells contain CaBP. Serum calcium concentrations were significantly elevated in 2 h and maximal by 12 h (an increase of 3.6 mg/dl). Calcium accumulation by the intestinal mucosa in vitro was increased by 6 to 8 h and maximal (60% increase over controls) at 24 h. Phosphate accumulation by the intestinal mucosa in vitro was increased by 6 h and maximal (105% increase over controls) between 8 and 24 h. 1,25(OH)2D3 increased CaBP and calcium accumulation by 4 h, 2 h sooner than did 1αOHD3. 1,25(OH)2D3 decreased serum calcium levels and increased serum phosphate levels at 2 h unlike 1αOHD3. No difference in the effects of these compounds on alkaline phosphatase activity, RNA polymerase activity, and phosphate accumulation could be demonstrated. These results are consistent with the possibility that 1αOHD3 may not require conversion to 1,25(OH)2D3 for all of its biological effects.
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  • 60
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    Keywords: Perifusion ; Parathyroid hormone ; In vitro ; Calcium ; Beta regulation
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    Notes: Summary An in vitro perifusion system was used to study parathyroid hormone (PTH) secretion in response to calcium (Ca) and beta-adrenergic agents. Perifused parathyroid tissue responded to changes in Ca within the physiologic range during experiments up to 5 h. There was rapid secretory stimulation after exposure to low Ca, with the maximum response being observed at 20 min. Normal bovine glands showed a Ca-independent nonsuppressible component of PTH release at concentrations of Ca above physiologic. 1-isoproterenol produced rapid stimulation of PTH release, the response being blocked by a beta antagonist. The maximum secretory response to either low Ca (0.5 mM) or 1-isoproterenol (10−5 M) was enhanced when the two stimuli were applied simultaneously. The response to isoproterenol was blocked by raising Ca to 2.5 mM. Although d,l-propranolol (10−4 M) caused mild suppression of PTH release at a Ca of 1.25 mM, it did not cause additional suppression at 2.5 mM Ca nor did it decrease the response to 0.5 mM Ca stimulation. The secretory response of the gland to low Ca was sustained at a level more than double the baseline rate. The response to isoproterenol was more transient, with a return to or toward baseline secretion within 60 min. These results suggest that beta agonists and low Ca have separate but related mechanisms for stimulating PTH release and may affect different pools of hormone. The perifusion system described is a relatively simple technique for assessing the kinetics and interactions of various stimulators of PTH secretion.
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    Calcified tissue international 32 (1980), S. 29-44 
    ISSN: 1432-0827
    Keywords: Rabbit ; Dentin ; Calcium ; Sulfur ; Periodicity ; Circadian ; Ultradian
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    Notes: Summary We have identified a variety of biological rhythms involved in the apposition and mineralization of dentin in the rabbit incisor. Animals were injected during the day or night with lead acetate at 2-week intervals—to provide biological time markers in forming dentin—and transverse undecalcified sections of the lower incisors were prepared for electron microprobe analysis. The positions of the lead markers were identified, and the continuous distribution of calcium and sulfur was measured at 1 µm intervals between the markers. In thin sections stained with hematoxylin after decalcification, the widths of a series of structural increments (bands) were measured with an ocular micrometer. Fourier analysis of the data revealed spectra of structural and compositional rhythms with a range of periodicities which extended from a matter of hours [ultradian (〈24 h)] to days [infradian (〉24 h) and circadian (approximately 24 h)]. The structural and compositional rhythms appeared to be independent to the extent that they did not necessarily have the same periods, or amplitudes. Nor were there simple phase relationships between all of the rhythms. At some times, Ca and S fluctuations are inversely proportional (180° out of phase), but in other cases they are directly proportional or out of phase by varying degrees other than 180°. The analyses thus suggest that calcium and sulfur deposition (representing mineral and glycosaminoglycan deposition, respectively) are not simply inversely proportional, and that the hematoxylin-stained structural increments did not solely reflect differences in the distribution of the mineral components in dentin.
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    Calcified tissue international 34 (1982), S. 253-257 
    ISSN: 1432-0827
    Keywords: Prednisolone ; Calcium ; Bone ; Corticoid osteopenia ; Vitamin D metabolites
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Adult male rats were fed a diet containing 0.15% calcium, 0.3% phosphorus, and either 100, 50, or 20 mg of prednisolone per kg of diet. All these levels of prednisolone led to osteopenia, decreased intestinal absorption of calcium, slightly lower serum calcium and phosphorus, and a decreased level of serum 1,25-dihydroxyvitamin D3. Exogenous parenteral 1,25-dihydroxyvitamin D3 corrected steroid-induced changes in serum calcium and phosphorus, but could not completely correct the low intestinal calcium transport; nor did it prevent the development of osteopenia. The prednisolone-induced osteopenia seems at least in part to be caused by impaired intestinal calcium transport. The impaired calcium transport may be the result of low levels of 1,25-dihydroxyvitamin D3 and a direct effect of presnisolone on the intestine.
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    Calcified tissue international 36 (1984), S. 596-603 
    ISSN: 1432-0827
    Keywords: Calcium ; Ameloblasts ; X-ray microanalysis ; Transport ; Frozen
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    Topics: Biology , Medicine , Physics
    Notes: Summary Energy dispersive X-ray microanalysis was applied to freeze-dried blocks of enamel organ tissue to determine levels of calcium in various celular regions. The tissue blocks were dissected free from adjacent forming enamel following injection of cobalt or fluoride ions, both of which temporarily inhibit enamel mineralization. In all control and experimental specimens there was an increasing gradient of calcium from the stratum intermedium cells to the distal ends of the ameloblasts. Calcium levels were significantly reduced near the distal ends of the ameloblasts following cobalt or fluoride injection as compared with controls. It is suggested that evidence of an intercellular buildup of calcium near the distal ends of the ameloblast supports a controlling function of these cells. The changes in calcium levels are correlated with alterations in mineralization known to occur in the adjacent enamel of the model systems employed.
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    Calcified tissue international 33 (1981), S. 477-484 
    ISSN: 1432-0827
    Keywords: 1,25-(OH)2-D ; Calcium ; Phosphorus ; Sex ; Age
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    Topics: Biology , Medicine , Physics
    Notes: Summary Dietary phosphate deprivation in women, but not men, is accompanied by a fall in plasma PO4 and a rise in plasma 1,25-(OH)2-vitamin D concentrations. In contrast, young male rats exhibit a fall in plasma PO4 and a rise in plasma 1,25-(OH)2-D concentrations in response to PO4 deprivation. To evaluate whether age and sex influence basal plasma 1,25-(OH)2-D levels and their regulation by PO4 deprivation, plasma 1,25-(OH)2-D, PO4, and Ca levels were measured in male and female rats ranging in age from 6 weeks to 6 months while they were eating normal or low PO4 diets for 1 to 16 days. Similar observations were also made in 6-week-old castrated male and female rats, males replaced with testosterone, and females replaced with estradiol. Basal plasma 1,25-(OH)2-D levels were higher in 6-week-old males (228±76 pmol/l) than in 6-week-old females (148±62 pmol/l;P〈0.01) and declined by age 11 weeks to stable levels averaging about 100 pmol/l without sex difference. Dietary PO4 deprivation resulted in a three-to fourfold increase in plasma 1,25-(OH)2-D concentrations regardless of age and sex, accompanied by a correlated rise in serum Ca concentrations. Castration of 6-week-old males and females eliminated the sex difference in basal plasma 1,25-(OH)2-D levels and appeared to enhance the elevation of plasma 1,25-(OH)2-D concentrations in response to PO4 deprivation in females. Although gonadal hormones may modify basal plasma 1,25-(OH)2-D levels, they are not required for the augmentation of plasma 1,25-(OH)2-D levels in response to PO4 deprivation.
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    Calcified tissue international 33 (1981), S. 575-581 
    ISSN: 1432-0827
    Keywords: Osteoporosis ; Vitamin D ; Bone mineral ; Estrogen ; Aging ; Thiazide ; Fluoride ; Calcium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Bone mineral content (BMC) was measured by125I photon absorptiometry every 3 months in 264 normal females (45–54 years) over a 2-year period together with serum samples for calcium, phosphate, magnesium, creatinine, alkaline phosphatases, potassium, and protein. A 48-h urinary calcium and creatinine measurement was obtained. The subjects were divided into 7 treatment groups and 3 placebo groups. Five of the treatments (thiazide, vitamin D3, fluoride + vitamin D3, fluoride, and 1αD3) were ineffective at the doses used; the annual loss of compact bone was 1.5–2.2% (-X=1.8%), similar to the loss seen with placebos. Estrogen and estrogen + thiazide, in contrast, produced a 1.34% annual increase of BMC. The subjects were divided into groups with low, medium, and high initial BMC. Also, individual regressions for bone change were calculated and the subjects were divided into groups of responders, maintainers, and losers (annual change of 〉0%, 0 to −1%, and 〉−1%, respectively). The initial BMC status did not consistently affect bone or biochemical responses to the therapeutic agents. Estrogen was effective even in subjects with high BMC, whereas the other agents did not inhibit bone loss even in subjects with low initial BMC. Virtually all subjects responded to estrogen positively; in contrast we could not identify a subset of “responders” with any of the other treatments. Time since menopause appeared to influence the bone changes, although it was not a significant effect given the sample size. Bone loss in groups not treated with estrogens was 2%/year at 20 months after menopause with a decline to 1.3%/year at 45 months post-menopause. There was no apparent decline in the bone response to estrogen during the first 4 years after menopause, and in fact bone response tended to increase with time.
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    Calcified tissue international 33 (1981), S. 485-488 
    ISSN: 1432-0827
    Keywords: 1,25-(OH)2-D ; Hypophysectomy ; Calcium ; Phosphorus
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    Topics: Biology , Medicine , Physics
    Notes: Summary The mechanism by which dietary phosphate deprivation elevates plasma 1,25-(OH)2-D levels is not known. To evaluate the role of the pituitary in regulating plasma 1,25-(OH)2-D concentrations, the responses of plasma 1,25-(OH)2-D to dietary phosphate deprivation and, separately, to dietary calcium deprivation were evaluated in intact and hypophysectomized male rats. Among intact and hypophysectomized rats eating normal diets, plasma 1,25-(OH)2-D levels averaged 228±76 and 148±62 pmol/1, respectively (P〈0.01). During dietary phosphate deprivation, plasma 1,25-(OH)2-D levels rose to 1160±260 in intact rats and fell to 90±26 pmol/l in hypophysectomized rats (P〈0.001). By contrast, during dietary calcium deprivation, plasma 1,25-(OH)2-D levels rose in both intact and hypophysectomized animals to 856±107 and 742±279 pmol/l, respectively (NS). In response to dietary phosphate deprivation, serum calcium concentrations rose as 1,25-(OH)2-D concentrations rose in intact rats but remained at control levels in hypophysectomized rats. These results support the hypothesis that a pituitary hormone acting either directly or indirectly on the kidney mediates the increase in plasma 1,25-(OH)2-D during dietary phosphate deprivation. The hypercalcemia that occurs in rats during dietary phosphate deprivation appears to depend on the elevation of plasma 1,25-(OH)2-D.
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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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    Calcified tissue international 24 (1977), S. 275-278 
    ISSN: 1432-0827
    Keywords: Calcium ; Thyroid ; Calcium ; Thyroïde
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    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les auteurs utilisent une solution de Glyoxal bis 2-hydroxyanil à basse température pour localiser le calcium dans la thyroïde. Grâce à cette technique qui limite les mouvements ioniques, ils observent que le calcium situé principalement dans la colloïde est résorbé en même temps qu'elle lors d'une stimulation par le froid.
    Notes: Summary With a Glyoxal bis 2-hydroxyanil alcoholic solution at −25°C, the calcium was localized in frozen thyroid slices. The limited ionic movement permits seeing that the calcium situated principally in the colloid is resorbed at the same time by a low temperature stress.
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    Calcified tissue international 30 (1980), S. 27-34 
    ISSN: 1432-0827
    Keywords: Ultrastructure ; Calcium ; Cartilage ; Vesicles
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    Notes: Summary The potassium pyroantimonate technique was utilized for the selective subcellular localization of calcium in the mandibular condylar cartilage of 1-day-old rats. Electron dense calcium pyroantimonate precipitates were localized principally in mitochondria and at the cell membrane of the chondrocytes. In addition, small intracellular vesicles 0.1–0.2µm in diameter were observed in proximity to the cell membrane of chondrocytes of the mid-hypertrophic zone. The results suggest that these vesicles were being extruded from the cell into the extracellular matrix. Energy-dispersive analysis by X-rays confirmed that calcium is the principal cation of the electron-dense precipitates.
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    Calcified tissue international 34 (1982), S. 43-48 
    ISSN: 1432-0827
    Keywords: Calcium ; Phosphate ; Phosphatidylserine ; Transport ; Mineralization
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    Topics: Biology , Medicine , Physics
    Notes: Summary Experiments were performed to study the effect of inorganic phosphate (Pi) on phosphatidylserine-mediated Ca2+ transport utilizing two- and three-compartment lipid-aqueous phase model systems. When the three-compartment model was used, the rate of Ca2+ transport from an aqueous donor compartment to an aqueous receiver compartment, separated by a nonaqueous phospholipid phase, was determined. This experiment showed that the rate of Ca2+ transport was proportional to the phosphatidylserine concentration and the pH. Pi modulated the rate of Ca2+ transport; even when the Pi concentration of the donor aqueous phase was low, there was a marked enhancement of transport. To determine whether the Pi-mediated rise in the Ca2+ transport rate was due to an increase in the uptake of Ca2+ into the lipid phase, or to an increase in the ability of the lipid phase to release Ca2+, a two-compartment model was used. It was found that the ability of the phosphatidylserine phase to take up Ca2+ increased as the Pi concentration of the aqueous donor phase was raised. With the increase in Ca2+ uptake there was a concomitant elevation in the rate of Ca2+ transport into an aqueous receiver phase. However, Pi did not stimulate Ca2+ release from the phosphatide. Thus it was concluded that Pi enhanced the interaction between Ca2+ and phosphatidylserine, possibly by forming a Ca-phospholipid-Pi complex. Once this interaction had taken place, Ca2+ release into the aqueous receiver compartment was independent of the Pi concentration.
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    Calcified tissue international 36 (1984), S. 77-82 
    ISSN: 1432-0827
    Keywords: Osteogenesis ; Diffusion chambers ; Alkaline phosphatase ; Calcium ; Phosphorus
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    Topics: Biology , Medicine , Physics
    Notes: Summary The alkaline phosphatase activity and the calcium and phosphorus content of osteogenic tissue formedin vivo following the implantation of diffusion chambers loaded with rabbit bone marrow cells is reported. (In this study the term osteogenic includes osteoblastic and chondroblastic.) Chambers examined 14–70 days after implantation revealed progressive accumulation of mineral. Alkaline phosphatase activity increased until day 30 and declined thereafter. The osteogenic potential of the marrow cells decreased with increasing weight (age) of the cell donor rabbit when measured either as the percentage of chambers containing osteogenic tissue or as the amount of calcium, phosphorus, or alkaline phosphatase activity within the chambers. The results confirm that measurements of these parameters in tissue formed by cells incubated in diffusion chambersin vivo may be used as a method for assay of osteogenesis.
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    Calcified tissue international 36 (1984), S. 269-273 
    ISSN: 1432-0827
    Keywords: Bone loss (osteopenia) ; Calcium ; Corticosteroids (glucocorticosteroids) ; Fluoride ; Vitamin D
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary To evaluate the effect of prednisone and triple treatment (sodium fluoride, calcium, and vitamin D) on trabecular and cortical bone serial bone mineral content (BMC) measurements were made at a metaphyseal (BMCD) and diaphyseal (BMCP) site on the forearm on 31 consecutive and previously bone-healthy patients scheduled for at least 24 weeks high-dose prednisone treatment. The patients were randomized into two further treatment groups: group I (n=16) received prednisone plus triple treatment and group II (n=15) received only prednisone. The two groups were similar with regard to age, sex, prednisone dose, and initial BMC. During 24 weeks treatment, BMCD (partially representing trabecular bone) and BMCP (mainly representing cortical bone) fell significantly and similarly, demonstrating that there is no preventive effect on bone mineral loss on the triple regimen. The BMC fall after 12 weeks was significantly more pronounced for metaphyseal (partially trabecular) than for diaphyseal (cortical) bone, whereas the values did not differ significantly after 24 weeks; this indicates a greater sensitivity to the hormone treatment of trabecular bone. In the entire group, the fall in BMC correlated positively with individual prednisone dose, significant at the diaphyseal site (r=0.39,P〈0.05), but not at the metaphyseal site (r=0.31, P=0.08). It is concluded that corticosteroid-induced osteopenia is a diffuse bone disease which affects trabecular as well as cortical bone, suggesting that BMC measured on the forearm reflects changes in bone mineral at other locations.
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    Calcified tissue international 36 (1984), S. 392-400 
    ISSN: 1432-0827
    Keywords: Vitamin D ; Chick embryo ; Bone ; Calcium ; Phosphate
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    Topics: Biology , Medicine , Physics
    Notes: Summary Chick embryos were injected in the yolk sac at various ages with various doses of different vitamin D3 metabolites. Serum concentrations of total calcium and inorganic phosphate were determined 24 h after the injection and histological and electron microscopic studies of the tibiae were conducted 3–6 days after. Confirming previous results, the injection of 1,25(OH)2D3 was found to produce significant hypercalcemia and hypophosphatemia. The dose required to produce these effects decreased with age: 100 ng on the 9th day, 50 ng on the 11th, and 10 ng on the 15th. This finding is interpreted as resulting from the fact that the specialized cells in the chorionic epithelium which are considered to be involved in mineral resorption from the shell differentiate between the 11th and 13th days. Although no bone changes were observed in embryos injected before the 11th day, a rim of unmineralized trabeculae (osteoid) was observed at the periphery of the cortex of the tibial diaphysis in the embryos which had been injected after that age. Thus, in embryos injected on the 11th day with 100 ng 1,25(OH)2D3, the trabeculae formed between the 11th and 14th day remained unmineralized until the 15th or 16th day at which time they completed their mineralization. In the embryos injected on the 14th day, the alterations were more severe and could be produced with doses 10 times smaller than those required when the injections were made on the 11th day. At all ages, the doses that produced an osteoid rim also induced hypercalcemia and hypophosphatemia. The electron microscopical study of the osteoid trabeculae showed that osteoblasts and osteocytes had normal cytological characteristics and that the bone matrix did not present changes other than the reduction in mineral deposition. While the above findings do not exclude a direct action of 1,25(OH)2D3 on bone cells as the mechanism of osteoid formation, they do underline the importance of the humoral changes at least as partial determinants of this phenomenon. The activities of various vitamin D metabolites were compared using as parameter the threshold-dose required to produce a rim of unmineralized trabeculae in the tibia of 14–15 days embryos (T-D). The most active metabolite appeared to be 1,25(OH)2D3 (T-D: 10 ng); it was followed by 1,24,25(OH)3D3 (T-D: 100 ng) and 1,25,26(OH)3D3 (T-D: 100 ng). Vitamin D3 itself (T-D: 100 µg), 25(OH)D3 (T-D: 2.5µg) and 24,25(OH)2D3 (T-D: 5 µg) produced similar responses but only when administered in much larger doses.
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    Calcified tissue international 36 (1984), S. 702-710 
    ISSN: 1432-0827
    Keywords: Corticosteroids ; Cartilage ; Organ culture ; Calcium ; Matrix vesicles
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The present study examined the effects of various corticosteroid and noncorticosteroid hormones upon the ultrastructure of chondroprogenitor cells and chondroblasts in an organ-culture system of late fetal condylar cartilage. Corticosteroids, (triamcinolone acetonide, dexamethasone, corticosterone) at concentrations of 10−6–10−8M stimulated markedly a precocious formation of matrix vesicles by chondroblasts. This stimulation was accompanied by a significant accretion of calcium complexes intra- and extracellularly in both the chondroprogenitor cell population and chondroblastsin vitro, as well as in the newly induced matrix vesicles. Nonglucocorticoid steroids (progesterone, estradiol, testosterone, cortexolone) did not evoke similar effects. Progesterone and testosterone, however, seemed to adversely affect the ultrastructure of the cartilage cells, whereas estradiol appeared to have a favorable effect on the morphology of cultured condylar cartilage.
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    Calcified tissue international 36 (1984), S. 550-555 
    ISSN: 1432-0827
    Keywords: Enamel crystals ; Length ; Shape ; Apatite ; Ultrastructure
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    Topics: Biology , Medicine , Physics
    Notes: Summary An original method for fractionating and preparing isolated crystals of homogeneous size was developed. It was demonstrated that enamel apatite crystals are at least 100 µm long. The flexibility of the very long crystallites was demonstrated. Crystal curvatures, accounting for the irregular course of the prisms through the enamel thickness, were visualized and measured. It was shown that in the deep forming enamel layer, lateral branches may grow out of the crystals and crystal fusing often occurs, inducing the crystallites to assume pyramidal shapes with their wide bases pointing toward the dentino-enamel junction and one or two tops toward Tomes' processes. During the maturation process, the two tops of the still immature crystals also fuse so that the mature crystals acquire a rodlike aspect, with parallel faces and steplike graduations along thec axis, allowing a close contact between the crystals. These results support the hypothesis that the crystallites would be continuous from the dentino-enamel junction to the surface.
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    Calcified tissue international 34 (1982), S. 121-124 
    ISSN: 1432-0827
    Keywords: Bone ; Calcium ; Metabolism
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    Topics: Biology , Medicine , Physics
    Notes: Summary Bone Gla protein (BGP) was measured in the plasma by radioimmunoassay (RIA) during treatment of 59 patients with bone diseases including Paget's disease (N=9), primary hyperparathyroidism (N=25), chronic renal failure (N=20), and cancer involving bone (N=5). Plasma BGP was increased above normal in all patients. BGP decreased in the patients with Paget's disease following the acute and chronic administration of salmon calcitonin. Plasma BGP was higher in women then in men with primary hyperparathyroidism. Following parathyroidectomy, BGP decreased in both sexes but the decrease was significant in women only. Plasma BGP was increased in patients with renal osteodystrophy and did not change after hemodialysis. In the patients with bone cancer, plasma BGP decreased during treatment of the attendant hypercalcemia with salmon calcitonin. Although plasma BGP and serum alkaline phosphatase (AP) levels were generally correlated in these studies, there were examples of dissociation between the two. The measurement of plasma BGP appears to provide a specific index of bone metabolism that may in some circumstances be more sensitive than serum alkaline phosphatase measurement. However, further studies are necessary to establish the clinical value of plasma BGP measurement by RIA in the management of patients with bone diseases.
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    Calcified tissue international 34 (1982), S. 382-390 
    ISSN: 1432-0827
    Keywords: Avian osteopetrosis ; Avian oncornavirus ; Ultrastructure ; Calcification ; Bone cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Diaphyseal tibial bone of 12.5 – 13-day and 19-day-old embryos and 20-day-old hatched chicks infected with retrovirus MAV.2-O were examined by transmission electron microscopy. The viruses were associated with lining osteoblasts and osteocytes. Whereas the infection of the osteoblast layer seemed to be a transient stage, virus association with osteocytes was a constant and main ultrastructural feature. The viruses were found either in the osteoid or in the periosteocytic space of the bone lacunae. They arose from dense cytoplasmic areas located near the cell plasmalemma via a budding process. The newly budded virus particles often had a large tail or a fine stalk-like process lost in the extracellular space. The viruses underwent calcification by deposition of inorganic material and were incorporated in the bone trabeculae. No production of virus was observed in typical osteoclasts with well-differentiated ruffled borders. The viral-induced avian osteopetrosis seemed to result from increased bone deposition through stimulation of osteoblast and osteocyte activities, whereas osteoclastic bone resorption seemed to be undisturbed.
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    Calcified tissue international 35 (1983), S. 107-110 
    ISSN: 1432-0827
    Keywords: Calcium ; Glucocorticoid ; Vitamin D ; Osteoporosis ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Eighty-eight adult male Sprague-Dawley rats were given a diet with either (a) 0.5% Ca and 0.6% P or (b) 0.01% Ca and 0.6% P. Osteopenia was created by adding prednisolone to the diet. The prophylactic effect of oral 1,25(OH)2D3 on the osteopenia was studied. It was found that prednisolone osteopenia in the rat was associated with defective Ca absorption. By giving an oral dose of 1,25(OH)2D3, it was possible to maintain normal Ca absorption during prednisolone treatment and to prevent the bone loss. No significant hypercalcemia or any kidney calcifications were seen. These results are in contrast to earlier findings, in which subcutaneous administration of 1,25(OH)2D3 failed to prevent prednisolone osteopenia because of its tendency to increase bone resorption.
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  • 79
    ISSN: 1432-0827
    Keywords: 31P NMR spectroscopy ; Phosphoprotein ; Dentin ; Calcium ; Inorganic orthophosphate ; Bovine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The single phosphoprotein of fetal calf dentin, having a molecular weight of approximately 94,000 and a phosphorus content of 8% (w/w), was examined by31P NMR spectroscopy. The single resonance at 3.7 ppm at pH 10 and its chemical shift during acid titration established the phosphomonoester nature of the organic phosphorus moiety. During titration of the phosphoprotein with CaCl2 in the presence of inorganic orthophosphate ions, line broadening for the orthophosphate resonance was both phosphoprotein- and calcium-dependent, indicating ternary complex formation. The data indicate that the phosphoprotein of fetal calf dentin binds both calcium and inorganic orthophosphate ions and therefore has the requisite physical chemical properties necessary for it to facilitate the heterogeneous nucleation of a Ca-PO4 solid phase from solution during tissue mineralization.
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  • 80
    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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  • 81
    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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  • 82
    ISSN: 1432-2048
    Keywords: Centrosema leghaemoglobin ; Nitrate reductase ; Nitrogen fixation ; Nitrogenase ; Vigna
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The sequence of events leading up to the establishment of symbiotic nitrogen-fixation were studied in two tropical legumes, Centrosema pubescens Benth, and Vigna unguiculata L. Walp. Parameters measured included fresh and dry weights, chlorophyll and leghaemoglobin contents, as well as the activities of NADH-nitrate reductase (EC 1.6.6.1), and nitrogenase (nitric-oxide reductase-EC 1.7.99.2) in plants that were inoculated with suitable rhizobia or which were watered with potassium nitrate. Dry weight and photosynthetic activity of both species followed the sigmoidal pattern which is characteristic of most plants. Growth was little different in either a qualitative or quantitative sense whether nitrogen was supplied as nitrate or through dinitrogen fixation. Although the biochemical sequence of events was dependent on the limiting sensitivities of the individual assays used, the data suggest that nitrate reductase is the first measurable enzymatic activity in the nodules (and roots), followed by acetylene reduction and leghaemoglobin in that order. It is possible therefore, that low levels of symbiotic nitrogen fixation occur in the nodules in the absence of leghaemoglobin. Nitrate reductase activity in C. pubescens nodules was negatively exponentially correlated with nitrogenase activity of the same nodules, suggesting a changing metabolism in old nodules. These data are discussed in terms of environmental and physical factors known to control nitrogen fixation.
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  • 83
    ISSN: 1432-2048
    Keywords: Leguminosae ; Nitrate-reductase mutants ; Nitrogen fixation ; Nodulation ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of nitrate on the symbiotic properties of nitrate-reductase-deficient mutants of a strain of cowpea rhizobia (32H1), and of a strain of Rhizobium trifolii (TA1), were examined; the host species were Macroptilium atropurpureum (DC.) Urb. and Trifolium subterraneum L. Nitrate retarded initial nodulation by the mutant strains to an extent similar to that found with the parent strains. It is therefore unlikely that nitrite produced from nitrate by the rhizobia, plays a significant role in the inhibition of nodulation by nitrate. Nitrite is an inhibitor of nitrogenase, and its possible production in the nodule tissue by the action of nitrate reductase could be responsible for the observed inhibition of nitrogen fixation when nodulated plants are exposed to nitrate. However, the results of this investigation show that nitrogen fixation by the plants nodulated by parent or mutant strains was depressed by similar amounts in the presence of nitrate. No nitrite was detected in the nodules. Nodule growth, and to a lesser extent, the nitrogenase specific activity of the nodules (μmol C2H4g−1 nodule fr. wt. h−1), were both affected by the added nitrate.
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  • 84
    ISSN: 1432-2048
    Keywords: Festuca ; Frost damage ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Tillers of Festuca arundinacea Schreb. were subjected to-8°C in a bath of methylated spirits for three-quarters of an hour. They were thawed at room temperature and some material taken from the shoot apical meristem and leaf blade for electron microscopy. Similar material was taken from control plants for electron microscopy. Nine tillers subjected to-8°C and thawed subsequently failed to regrow. Nine control tillers regrew. All the treated meristem cells and about half the treated leaf mesophyll cells were extensively altered. Their nuclei were contracted, organelles were swollen or partly disrupted, plasmalemma and nuclear membranes were broken or absent and vacuoles were sometimes disrupted. Strongly osmiophilic material accumulated in the vicinity of membranes. About half the leaf mesophyll cells differed from the control mesophyll cells only in having more spherosomes and narrower thylakoids. Parallels with other ultrastructural studies of stress damage and the indications the results give of possible primary damaging events are discussed.
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  • 85
    ISSN: 1432-2048
    Keywords: Ammonium assimilation ; Glycine ; Nitrogen fixation ; Proplastid ; Purine synthesis ; Root nodule ; Ureide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Subcellular organelle fractionation of nitrogen-fixing nodules of soybean (Glycine max (L.) Merr.) indicates that a number of enzymes involved in the assimilation of ammonia into amino acids and purines are located in the proplastids. These include asparagine synthetase (EC 6.3.1.1), phosphoribosyl amidotransferase (EC 2.4.2.14), phosphoglycerate dehydrogenase (EC 1.1.1.95), serine hydroxymethylase (EC 2.1.2.1), and methylene-tetrahydrofolate dehydrogenase (EC 1.5.1.5). Of the two isoenzymes of asparate aminotransferase (EC 2.6.1.1) in the nodule, only one was located in the proplastid fraction. Both glutamate synthase (EC 1.4.1.14) and triosephosphate isomerase (EC 5.3.1.1) were associated at least in part with the proplastids. Glutamine synthetase (EC 6.3.1.2) and xanthine dehydrogenase (EC 1.2.1.37) were found in significant quantities only in the soluble fraction. Phosphoribosylpyrophosphate synthetase (EC 2.7.6.1) was found mostly in the soluble fraction, although small amounts of it were detected in other organelle fractions. These results together with recent organelle fractionation and electron microscopic studies form the basis for a model of the subcellular distribution of ammonium assimilation, amide synthesis and uredie biogenesis in the nodule.
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  • 86
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    Planta 146 (1979), S. 31-39 
    ISSN: 1432-2048
    Keywords: Autoradiography ; Calcium ; Mougeotia ; Phytochrome
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Ca2+ is proposed to function as a messenger in such phytochrome-mediated responses as localized cell growth, intracellular movements, and control of plasma membrane properties. To test this hypothesis, the uptake of Ca2+ in irradiated and non-irradiated regions of individual threads of the green alga Mougeotia was studied with the aid of 45Ca2+ and low temperature autoradiography: 10–20 cells within 40–60 cell-long threads were irradiated for up to 1 min, transferred to darkness for 3 to 10 min, submersed in a radioactive medium for 1 min, washed in an unlabelled medium for 30 min, and then autoradiographed at-80° C for several days. The autoradiographs show that those cells which had been pre-irradiated with red light did take up 2–10 times more Ca2+ than the adjacent non-irradiated cells of the same thread. Cells pre-irradiated with farred light or red light followed by far-red light showed no enhanced uptake of Ca2+. These results might be interpreted to indicate, firstly, that phytochrome-Pfr is involved in the enhanced uptake of Ca2+ and secondly, that the accumulation of radioactive Ca2+ in red light irradiated cells is an expression of an increased intracellular concentration of Ca2+. This interpretation is based on the data that (i) the dark interval between irradiation and labelling precluded the involvement of photosynthesis, (ii) the effect of red light was reversible with far-red light, and (iii) the accumulation of Ca2+ persisted during the long wash-out period. We speculate, that the red light-enhanced accumulation of Ca2+ in Mougeotia cells is caused by a Pfr-mediated increase of the Ca-permeability of the plasma membrane, and perhaps by a Pfr-impeding of an active Ca2+-extrusion.
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  • 87
    ISSN: 1432-2048
    Keywords: Calcium ; Cation exchange ; Lycopersicon esculentum ; Rhythm ; diurnal Transport (calcium) ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Tomato (Lycopersicon esculentum Mill. cv. Moneymaker) and maize (Zea mays L. cv. spec.) plants were supplied with 45Ca-labeled nutrient solutions for a period of 8 or 16 h in the dark, in the light, or in a light-dark régime. Plant parts were analyzed for 45Ca content. The partitioning of 45Ca between mature leaves and meristems was shown to be affected by the presence of light. The transport of 45Ca to meristems was higher in a dark period than in a comparable light period. Experiments with excised tomato shoots yielded similar distribution patterns of 45Ca over leaves and meristems, thus excluding root pressure as the main driving force for the enhanced import of 45Ca into the meristems in the dark. Results are discussed in terms of cation-exchange transport and competition between the various calcium sinks.
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  • 88
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    Planta 158 (1983), S. 320-328 
    ISSN: 1432-2048
    Keywords: Calcium ; Egeria ; Fluorescent probe ; Ions, group II ; Symplast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The hydrophyllic dyes fluorescein glutamic acid, fluorescein glutamylglutamic acid (F(Glu)2), fluorescein hexaglycine, fluorescein leucyldiglutamyl-leucine and 6-carboxyfluorescein are unable to pass the plasmalemma in leaves of E. densa. However, when injected into single cells the dye conjugates of molecular weight 665 dalton or less move freely from cell-to-cell. This intercellular movement presumably occurs via the plant symplast. Movement of F(Glu)2 from the injected cell occurs with greatly reduced frequency when Ca2+, Mg2+ or Sr2+ are injected into the cell immediately prior to the dye. The fraction of dye injections leading to movement declines with increasing group II ion concentration in the electrode tip, up to 10 mM. Sodium and K ions do not affect dye movement. When dye injection is delayed 30 min after Ca2+ injection, dye movement is no longer inhibited. Thus the cells recover from the Ca2+ injection, indicating that the ion does not cause major cell damage. Recovery from Mg2+ injection is not complete within 60 min. Treatment of leaves with chemicals expected to raise the concentration of free intracellular group II ions, notably the mitochondrial uncoupler carbonyl cyanide p-trifluoromethoxyphenyl hydrazone, the inhibitor of mitochondrial Ca2+ uptake trifluralin, or the ionophore A23187 also inhibits dye movement, while the calmodulin inhibitor trifluoperazine does not. Cytoplasmic streaming is inhibited by Ca2+ or Mg2+ injection and by the metabolic inhibitors. However when streaming is stopped by cytochalasin B, dye movement is not inhibited. Hence steaming is not necessary for dye movement. Thus the cytoplasmic concentration of free group II ions may directly regulate the permeability of the plant symplast.
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  • 89
    ISSN: 1432-2048
    Keywords: Bryophyta ; Bud formation (moss) ; Calcium ; Chlorotetracycline ; Cytokinin ; Funaria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have investigated the changes in membrane-associated calcium that occur during cytokinin induced bud formation in Funaria hygrometrica Hedw. using the fluorescent Ca2+-chelate probe chlorotetracycline (CTC). In the target caulonema cells a localization of CTC fluorescent material becomes evident at the presumptive bud site 12 h after cytokinin treatment. By the time of the initial asymmetric division this region is four times as fluorescent as the entire caulonema cell. Bright CTC fluorescence remains localized in the dividing cells of the bud. To relate the changes in CTC fluorescence to changes in Ca2+ as opposed to membrane-density changes we employed the general membrane marker N-phenyl-1-naphthylamine (NPN). NPN fluorescence increases only 1.5 times in the initial bud cell. We conclude that the relative amount of Ca2+ per quantity of membrane increases in this localized area and is maintained throughout bud formation. We suggest that these increases in membrane-associated Ca2+ indicate a localized rise in intracellular free Ca2+ concentration brought about by cytokinin action.
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  • 90
    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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  • 91
    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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  • 92
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    Planta 144 (1979), S. 401-406 
    ISSN: 1432-2048
    Keywords: A23187 gradients ; Calcium ; Germination (moss spores) ; Electric fields-Funaria ; Ionophore gradient ; Moss spores-Polarization ; Spores (moss)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have used both steady electric fields, and gradients of the divalent ionophore, A23187, to control the point at which rhizoids emerge from spores of the common moss Funaria hygrometrica. The spores were grown in a medium containing calcium nitrate as the only major salt. Spores tend to form rhizoids towards the positive electrode, with a half maximal response to a difference of 4–8 mV across each cell. They also tend to form rhizoids towards the end of higher ionophore concentration in response to A23187 gradients. Both of these responses are the same at pH 5.5 and 8.0. Our tentative explanation is that Funaria spores tend to form rhizoids where most calcium enters. However, the point of chloronema emergence is scarcely affected by steady fields of up to 45 mV/cell. Moreover, when steady fields are applied across already developed rhizoids or chloronemata, their subsequent growth is directed towards the negative electrode in both cases, with rhizoids giving a 50% response at only 3—5 mV/cell, and chloronemata being less responsive.
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  • 93
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    Planta 152 (1981), S. 450-456 
    ISSN: 1432-2048
    Keywords: Calcium ; Cell elongation ; Gibberellin ; Lactuca ; pH and growth
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The relationship between calcium ions and gibberellic acid (GA3)-induced growth in the excised hypocotyl of lettuce (Lactuca sativa L.) was investigated. The short-term kinetics of growth responses were measured using a linear displacement transducer. Test solutions were added either as drops to the filter paper on which the hypocotyl stood (“non-flow-past”) or by switching solution flowing past the base of hypocotyl (“flow-past”), resulting in differences in growth behavior. Drops of CaCl2 added at a high concentration (10 mM) inhibited growth within a few minutes. This inhibition was reversed by ethylenediaminetetraacetic acid (EDTA). Drops of EDTA or ethyleneglycol-bis(2-aminoethylether)-tetraacetic acid caused a rapid increase in growth rate. Growth induced by EDTA was not further promoted by GA3. A continuous H2O flow resulted in growth rates comparable to those in response to GA3. Addition of CaCl2 to the flow-past medium inhibited growth and this inhibition was reversed by a decrease in CaCl2 concentration. The growth rate was found to be a function of CaCl2 concentration. When a constant CaCl2 concentration was maintained by the flow-past medium, a shift in pH from 5.5 to 4.25 had no obvious effect on hypocotyl elongation. Gibberellic acid was found to reverse the inhibitory effect of CaCl2, causing an increase in growth rate similar to that found previously when GA3 was added to hypocotyls grown in H2O under non-flow-past conditions. We propose that gibberellin controls extension growth in lettuce hypocotyl sections by regulating the uptake of Ca2+ by the hypocotyl cells.
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  • 94
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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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  • 95
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    Planta 133 (1976), S. 15-19 
    ISSN: 1432-2048
    Keywords: Tropaeolum, Embryogenesis ; Differentiation ; Plastids ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Embryogeny in the nasturtium is characterized by the development of a large, tripartite suspensor and storing cotyledons. A light and electron microscopic study revealed an early diversification of the plastids in the various regions of the suspensor and the embryo proper. Amyloplasts are found in the developing cotyledons of the heart-like embryo, while chloroplasts occur within the meristematic part of the embryo and the adjacent portion of the suspensor. The cells between the meristem and the storing cotyledons display undifferentiated leukoplasts, whereas leukoplasts with an electron-dense matrix occur in the basal cell mass of the embryo-suspensor. Etioplasts develop in several cells of the placental haustorium of the suspensor. The carpel haustorium shows rather undifferentiated leukoplasts, which are transformed into electron-dense plastids during autolysis of the suspensor. This early plastidal differentiation in discussed with respect to its control and functional significance.
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  • 96
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    European biophysics journal 3 (1977), S. 55-58 
    ISSN: 1432-1017
    Keywords: Rhodopsin ; Rods ; Phototransduction ; Calcium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Physics
    Notes: Abstract Difficulties in testing the possible role of calcium as a Transmitter in the R.O.S. are discussed. A comparison is made with the Sarcoplasmic Reticulum system where calcium flux are easily measured. The latest results reviewed on intact cellular structures are highly indicative but not yet conclusive.
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  • 97
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    European biophysics journal 3 (1977), S. 141-143 
    ISSN: 1432-1017
    Keywords: Calcium ; Limulus ; Photoreceptors ; Adaptation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Physics
    Notes: Abstract Calcium ion fulfills several criteria for identifying an intracellular messenger for light-adaptation in Limulus photoreceptors. Direct injection of Ca++ mimicks two aspects of light-adaptation; sequestration of intracellular calcium tends to prevent light-adaptation; and light induces an increase in intracellular Ca++ as demonstrated by two independent techniques.
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  • 98
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    European biophysics journal 3 (1977), S. 145-151 
    ISSN: 1432-1017
    Keywords: Visual cell membranes ; Invertebrates ; Ion channels ; Activation of light channels ; Calcium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Physics
    Notes: Abstract It is assumed that “dark channels” determine a permanent dark conductance of the arthropod visual cell membrane. The light stimulus causes a transient opening of “light channels”. The ion selectivity of dark channels and light channels is roughly described. Factors influencing the activation of light channels, as membrane energy metabolism, membrane potential and adjusted calcium ion concentration are specified. The mechanism of the action of calcium ions on the conductance of the visual cell membrane is discussed.
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  • 99
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    European biophysics journal 5 (1979), S. 33-41 
    ISSN: 1432-1017
    Keywords: Rhodopsin ; Calcium ; Reaction kinetics ; Regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Physics
    Notes: Abstract The formation of metarhodopsin II in various bovine rhodopsin preparations (rod outer segment (ROS) suspensions and rhodopsin-detergent solutions) was measured by means of flash spectrophotometry. The half-lifetime and formation of metarhodopsin II in ROS did not depend on the calcium concentration in the range of less than 10−9 M (using EGTA or EDTA) to 15×10−3 M calcium at pH values of 5.0, 7.1, and 9.0 (Table 1). The regeneration of rhodopsin from opsin by adding 11-cis retinal to ROS-suspensions and rhodopsin digitonin solutions was measured spectrophotometrically. It was not substantially different in either saline, one containing less than 10−7 M calcium (by adding EGTA), the other containing 10−3 M calcium (Table 2).
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  • 100
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    European biophysics journal 5 (1979), S. 223-230 
    ISSN: 1432-1017
    Keywords: Prolonged depolarising after-potential (PDA) ; Visual pigment ; Invertebrate photoreceptor ; Manganese ; Calcium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Physics
    Notes: Abstract We have studied the effects on the PDA of modifying intracellular and extracellular concentrations of Ca2+ and Mn2+. The effect of decreased Ca2+ concentration or addition of EGTA is mainly an increase in the PDA amplitude and length. Raising Ca2+ concentration using ruthenium red or high external Ca2+ has the opposite effect. The effect of Mn2+ is much more striking: In the presence of 50–100 mM Mn2+ the PDA is initially greatly depressed but can rise slowly for up to 20 or 30 s (in the dark) until it approaches its original amplitude and time course. Bridge measurements showed that the depression of the PDA corresponds to a depressed conductance and so is not due to an increase in K+ conductance. The Mn2+ effect is potentiated by decreased Ca2+. Appropriate stimulation suppresses the rising PDA as promptly as it does a normal PDA, suggesting that if lateral diffusion is the source of the slow rise, the PDA and PDA-depressing processes must be spatially linked. The action of the anti-PDA is apparently prolonged by both Ca2+ and Mn2+.
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