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  • Electron microscopy  (168)
  • Calcium  (114)
  • Springer  (281)
  • Annual Reviews
  • 1980-1984  (257)
  • 1965-1969  (24)
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  • 1
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    Springer
    Colloid & polymer science 261 (1983), S. 373-374 
    ISSN: 1435-1536
    Keywords: Electron microscopy ; short-time staining ; nodular structure ; crystallization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
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  • 2
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    Colloid & polymer science 260 (1982), S. 564-569 
    ISSN: 1435-1536
    Keywords: lin. Polyethylene ; Single crystals ; Heat of Fusion ; DSC ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Notes: Abstract Recently published results for solution crystallized PE single crystals have shown, that the experimental heat of fusionΔH * is higher, if the solvent is exchanged to silicon oil (oil suspension samples) as compared with dried mats. This has been interpreted by the collapse of the original hollow pyramids during drying, inducing lateral defects within the lamellae. The present investigation does not confirm this unexpected result.ΔH * of dried mats (T c 66 to 91 °C) and of the corresponding oil suspension samples agree within the rather small limits of experimental error. The crystallinities as derived fromΔH *, density or WAXS are in excellent agreement. SEM micrographs of cold fractured dried mats show their spongy macromorphology, but TEM micrographs of stained ultra-thin sections reveal the lamellar morphology of the walls, consisting of curved lamellae and stacked hollow pyramides. If a dried mat is sintered at room temperature, a dense transparent film is obtained with a rather regular stacked morphology of large flat lamellae.ΔH * of these films agrees with that of the original mat.
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  • 3
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    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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  • 4
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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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  • 5
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    Calcified tissue international 3 (1969), S. 103-105 
    ISSN: 1432-0827
    Keywords: Mitochondrion ; Calcium ; Excretion ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
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  • 6
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    Calcified tissue international 2 (1968), S. 296-298 
    ISSN: 1432-0827
    Keywords: Calcium ; Lipid ; Bacteria ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé On a fait ce travail pour determiner si le facteur responsable pour la liaison de calcium par un calcifableBacterionema matruchotii est dans la fraction lipide de la cellule. Des cellules congelees et sechees ont ete extraites par le chloroform-methanol. La fraction de chloroform-methanol, les cellules extraites et les cellules non traitees ont ete examinees pour la liaison de calcium. La fraction du chloroform-methanol et les cellules non traitees avaient la liaison de calcium. Les cellules extraites n'en avaient pas.
    Abstract: Zusammenfassung Diese Arbeit wurde durchgeführt um festzustellen, ob sich der Faktor für die Calcium-bindung, durch das calcifizierendeBacterionema matruchotii, in der Lipoidfraktion befindet. Die lyophiilisierten Zellen wurden mit Chloroform-Methanol extrahiert. Die Chloroform-Methanol-Fraktion, die extrahierten Zellen, sowie die nicht behandelten Zellen wurden auf eine Calciumbindung hin untersucht. Die Chloroform-Methanol-Fraktion und die nicht behandelten Zellen demonstrierten eine Calciumbindung. Die extrahierten Zellen hingegen nicht.
    Notes: Abstract This work was done to determine whether the factor responsible for calcium binding by a calcifiableBacterionema matruchotii is in the lipid fraction of the cell. Freeze-dried cells were extracted with chloroform-methanol. The chloroform-methanol fraction, the extracted cells and untreated cells were examined for calcium binding. The chloroform-methanol fraction and the untreated cells bound calcium. The extracted cells did not.
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  • 7
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    Calcified tissue international 3 (1969), S. 340-347 
    ISSN: 1432-0827
    Keywords: Bone ; Calcium ; Fluoride ; Ingestion ; compatibility
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Des expériences ont été faites sur le rat afin de tester les possiblités d'ingestion perorale simultanée de fluor (F) et de calcium (Ca) en proportions calculées pour traitement de certaines ostéopénies humaines. F sous forme de Na2PO3F et Ca sous forme de gluconate de calcium (CaGluc) n'influençaient pas l'un l'autre quant à l'utilisation par le squelette (fémur). Une viscosité élevée, produite par l'addition d'amidon ou de cellulose carboxyméthylée (CMC) à la solution ou dilution ingérée, augmentait l'utilisation du F même si CaGluc était remplacé par citrate de calcium, qui en soi avait un effet réducteur modéré sur l'utilisation du F. Le glycérophosphate de calcium réduisait fortement l'utilisation du F même en présence de CMC. L'utilisation du F comme NaF était fortement réduite par CaGluc, même en présence de CMC. Les concentrations testées de Na2PO3F, NaF ou CMC n'influençaient pas l'utilisation squelettique de Ca comme CaGluc.
    Abstract: Zusammenfassung Es wurden Rattenexperimente mit markierten Substanzen durchgeführt, um die Auswirkung simultaner peroraler Gaben von Fluor (F) und von Calcium (Ca) zu prüfen, und zwar in einem Verhältnis, das für die Behandlung gewisser menschlicher Osteopenien berechnet wurde. Fluor in Form von Na2PO3F und Ca in Form von Calciumgluconat (CaGluc) interferieren gegenseitig nicht bei der Verwertung durch das Skelet (Femur). Eine hohe Viscosität der eingegebenen Lösung oder der Aufschlämmung, die durch Zusatz von Stärke oder Carboxymethylcellulose (CMC) erzielt wurde, erhöhte die Verwertbarkeit von F sogar wenn CaGluc durch Calciumcitrat ersetzt wurde, welches die F-Verwertung leicht reduzierte. Calcium-glyzerophosphat verminderte die Fluoraufnahme in den Knochen stark, sogar in Anwesenheit von CMC. Die Verwertung von F als NaF war stark herabgesetzt durch CaGluc, selbst beim Vorhandensein von CMC. Die untersuchten Konzentrationen von Na2PO3F, NaF oder CMC hatten keinen Einfluß auf die Calciumaufnahme im Skelet in Form von CaGluc.
    Notes: Abstract Rat experiments with labelled compounds were carried out in order to test the possibilities of simultaneous peroral supply of fluorine (F) and calcium (Ca) in proportions calculated for treatment of certain human osteopenias. F in the form of Na2PO3F and Ca in the form of calcium gluconate (CaGluc) did not interfere with each other's utilisation by the skeleton (femur). A high viscosity produced by adding starch or carboxymethyl cellulose (CMC) to the ingested solution or slurry increased the utilisation of F even when CaGluc was replaced by calcium citrate, which moderately reduced F utilisation. Calcium glycerophosphate strongly reduced F utilisation even in the presence of CMC. The utilisation of F as NaF was strongly reduced by CaGluc, even in the presence of CMC. The tested concentrations of Na2PO3F, NaF or CMC did not influence the skeletal utilisation of Ca as CaGluc.
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  • 8
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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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  • 9
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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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  • 10
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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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  • 11
    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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  • 12
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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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  • 13
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    Calcified tissue international 4 (1969), S. 224-230 
    ISSN: 1432-0827
    Keywords: Calcinosis ; Calcium ; Iron ; Phosphate ; Collagen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé La calcification de la peau (calciphylaxie) provoquée par l'administration sous-cutanée de chlorure de fer chez des rats sensibilisés par le dihydrotachystérol (DHT) est précédée par une déposition sélective de fer (décelable à la microscopie électronique) à la surface des fibres collagènes. Les analyses à la microsonde prouvent que les taux de calcium et de phosphore sont augmentés à l'endroit de l'injection du chlorure de fer même sans l'administration préalable de DHT; cependant, l'apatite ne se forme que chez les animaux prétraités par le DHT. La quantité et le degré de cristallinité de la matière inorganique sont presque identiques au sixième et au trentième jour de l'expérience.
    Abstract: Zusammenfassung Durch elektronenmikroskopische Untersuchungen wurde festgestellt, daß die durch subcutane Verabreichung von Eisenchlorid bei Dihydrotachysterin-sensibilisierten Ratten herbeigeführte Hautverkalkung durch eine selektive Eisenablagerung auf der Oberfläche der Kollagenfibrillen eingeleitet wird. Die Röntgenmikroanalyse zeigt an, daß die lokale Calciumund Phosphorkonzentration bereits nach Eiseninjektion allein ansteigt, während Apatitbildung nur in den mit DHT vorbehandelten Tieren erfolgt. Ausmaß und Menge der Kristallisation sind am 6. und 30. Tage nach Versuchsbeginn fast gleich.
    Notes: Abstract Calcification of the skin (calciphylaxis) induced by the subcutaneous administration of iron chloride to dihydrotachysterol (DHT)-sensitized rats is preceded by the selective deposition of iron (visible with electron microscopic techniques) on the surface of collagen fibrils. Electron probe X-ray microanalysis indicates that the injectionof iron alone suffices to increase local concentrations of calcium and phosphorus; however, apatite formation occurs only in animals pretreated with DHT. The amount and degree of crystallinity of the inorganic material is very similar on the sixth and thirtieth day after the beginning of the experiment.
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  • 14
    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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  • 15
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    Development genes and evolution 191 (1982), S. 205-207 
    ISSN: 1432-041X
    Keywords: Chitin inhibition ; Nikkomycin ; Cuticle ; Electron microscopy ; Epilachna varivestis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The nucleoside antibiotic nikkomycin has proved to be an effective inhibitor of chitin synthesis in the Mexican bean beetleEpilachna varivestis. Ultrastructural investigations show defects in the procuticular area after nikkomycin application which suggest the complete absence of chitin. A cuticle like this is inflexible and too brittle to satisfy its normal function as an exoskeleton. The individuals are not able to free themselves from the exuvia and finally die. Therefore nikkomycin seems to be a potential insecticide with high specifity.
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  • 16
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    Calcified tissue international 33 (1981), S. 143-151 
    ISSN: 1432-0827
    Keywords: Calcium transport ; Cytochalasin B ; Dihydrocytochalasin B ; Colchicine ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary In vivo calcium absorption was studied in normal and rachitic chicks. Cytochalasin B (CB) at a concentration of 25 µg/ml added to the medium inside the duodenal lumen inhibited calcium absorption (20 min) from 82.5±1.9% of calcium absorbed in the controls to 59.2±3% in normal and from 70.0±2.3% to 47.0±2.1% in rachitic chicks. In vitro studies by everted ileal sacs of young rabbits also showed an inhibition of active transport of calcium due to CB. Whereas in the controls the ratio of45Ca concentrations in serosal and mucosal media (60 min) was 7.2±0.32, the ratios were 5.24±0.52; 4.40±0.36; 3.40±0.42; 5.77±0.52; 1.38±0.08; and 1.06±0.02 in the presence of CB at concentrations of 5, 10 and 25 µg/ml; colchicine 10−4M, Na citrate 0.02M, and heat-devitalized conditions, respectively.45Ca concentration in the mucosal scrapings was also affected. It showed an increase from controls (15,101±404 cpm/mg) and correlated with CB concentration: 17,378±489, 19,015±1000, and 20,201±362 at 5, 10, and 25 µg/ml, respectively. Dihydrocytochalasin B also inhibited active calcium transport and caused an increase in45Ca concentration in the mucosal scrapings. Correlated electron microscopic studies showed certain changes in the brush border, especially in some actin microfilaments in the terminal web region. It seems that these morphological alterations may be related to transcytoplasmic movement of calcium.
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  • 17
    ISSN: 1432-0827
    Keywords: Alcohol ; Electron microscopy ; Growth plate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary We have previously demonstrated that ethanol has a direct toxic effect on the rat skeleton characterized by decreased trabecular bone volume. In the present study, we examined the ultrastructure of the distal radial epiphyseal growth plates in these same animals. Eight weeks of ethanol administration to 12 male rats results in serum alcohol levels of 140 mg/dl but did not alter the width or light microscopic appearance of the radial growth plate. Quantitative electron microscopy failed to demonstrate morphologic evidence of toxicity in the skeletal cells. We conclude that although ethanol appears to have a direct effect on rat bone characterized by enhanced resorption, toxicity is not attended by ultrastructural changes in the skeletal cells.
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  • 18
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    Calcified tissue international 33 (1981), S. 529-540 
    ISSN: 1432-0827
    Keywords: Bone cells ; Electron microscopy ; PTH ; PGE1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Four different cell populations—designated PF, OB, OC, and PC—were isolated from calvaria of 18-day-old chick embryos for analysis of the effects of hormones on bone tissue. The cell populations were studied with histological and biochemical methods. Apart from the well-known cell types present in calvaria, a new cell type was found in the noncalcified organic matrix between the osteoblastic layer and the calcified matrix. These cells were provisionally called osteocytic osteoblasts. They represent the “transition state” between osteoblasts and osteocytes. On the basis of histological studies with light microscopy (LM), transmission electron microscopy (TEM) and scanning electron microscopy (SEM), the PF population was considered to originate primarily from the periosteal fibroblasts, the OB population from the osteoblasts and osteocytic osteoblasts. The population of cells still present in calvaria after removal of periosteal fibroblasts and osteoblasts was called the OC population. This cell population was very much enriched with osteocytes. The fourth isolated population (PC) was a mixed population of fibroblasts, osteoblasts, and preosteoblasts. On exposure to parathyroid hormone (PTH), all four cell populations showed increased lactate production, but only the OB and OC populations displayed increased cAMP production. Prostaglandin E1 (PGE1) stimulated cAMP production in both OB and PF cells. From the results of this study it was concluded that PTH receptors are present on all of the cell types studied, but that occupancy of the receptor induces adenylate cyclase stimulation only in osteocytes and fully differentiated osteoblasts.
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  • 19
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    Calcified tissue international 3 (1969), S. 184-193 
    ISSN: 1432-0827
    Keywords: Mitochondria ; Calcification ; Calcium ; Chondrocytes ; Growth plate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Des cultures d'épiphyse prélevées sur des rats et des souris ont été étudiées sous le microscope électronique. Un profile de granules mitochondriques de densité électronique a été trouvé. Les chondrocytes dans la zone proliférative avaint peu de granules, alors que ceux des zones successives ont montré une augmentation progressive de leur nombre et densité jusqu'à ce que la zone de calcification provisoire ait été atteinte. Cette zone a montré une distribution périphérique de mitochondries et une réduction du nombre et de la densité des granules mitochondriques. Du calcium isotopique 47 a été utilisé autoradiographiquement pour déterminer la location de calcium dans ces cellules. Des grains ont été trouvés sur les membranes R.E. et sur la plupart des mitochondries. La preuve d'un profile de ces granules et de leur rapport spatial avec la face de minéralisation indique une action éventuelle de mitochondries dès le début de la calcification de la matrice.
    Abstract: Zusammenfassung Epiphysekulturen von Ratten und Mäusen wurden unter dem Elektronenmikroskop untersucht. Ein Profil von mitochondrischen Körnchen mit elektronischer Dichte wurde gefunden. Chondrozyten in der Proliferationszone wiesen wenig Körnchen auf, während die der nachfolgenden Zonen allmählich an Zahl und Dichte zunahmen, bis die Zone der provisorischen Verkalkung erreicht wurde. Diese Zone zeigte eine periphere Verteilung der Mitochondrien und eine Abnahme in Zahl und Dichte der mitochondrischen Körnchen. Isotopes Kalzium 47 wurde autoradiographisch verwendet, um die Lage des Kalziums in diesen Zellen zu bestimmen. Körnchen wurden auf den E.R.-Membranen und auf einem Großteil der Mitochondrien gefunden. Der Nachweis eines Profils dieser Körnchen und ihres räumlichen Verhältnisses zur Mineralisierungsfläche weist auf einen möglich Einfluß der Mitochondrien mit Beginn der Matrixverkalkung hin.
    Notes: Abstract Rat and mice epiphyseal growth plates were studied with the electron microscope. A gradient of mitochondrial electron-dense granules was found. Chondrocytes in the proliferative zone had few granules, while those of the succeeding zones showed a gradual increase in number and density until the zone of provisional calcification was reached. This zone showed a peripheral distribution of mitochondria and a decrease in the number and density of mitochondrial granules. Isotopic47calcium was used autoradiographically to determine the location of calcium in these cells. Grains were found over the endoplasmic reticulum membranes and over most mitochondria. The demonstration of a gradient of these granules and their spatial relation to the mineralization front suggests a possible involvement of mitochondria in the onset of matrix calcification.
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  • 20
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    Calcified tissue international 30 (1980), S. 43-50 
    ISSN: 1432-0827
    Keywords: Osteogenesis ; In vitro ; Electron microscopy ; Mineralization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Chick limb mesenchymal cells differentiate into muscle, cartilage, fibrous, and bone tissue. Previous reports show that when stage 24 limb mesenchymal cells are cultured in vitro, chondrocytes, myocytes, fibrocytes, and osteoblasts can be identified on the basis of morphological and biochemical parameters. The study reported here demonstrates that phenotypic expression in culture seems to be dependent on the initial plating density, Scanning electron microscopic observations indicate that when stage 24 limb mesenchymal cells are initially seeded at high densities (5 × 106 cells per 35 mm culture dish), mounds of cells appear in culture. These mounds represent cartilage nodules composed of a fine fibrous matrix and chondrocytes, surrounded by a loose fibrous connective tissue matrix. Cultures initially plated at intermediate densities (2.0–2.5 × 106 cells/35 mm culture dish) produce a flattened layer of fibrocytes overlying a matrix of collagen fibers and calcium phosphate deposits as determined by electron-microprobe analysis; these observations are indicative of osteoblast expression. Cells seeded at this intermediate density appear larger and possess greater surface area than cells seeded at high density. It is suggested that conditions that permit such increased cell surface area coupled with a relative compaction due to cell crowding may provide conditions permissive for osteogenesis. Based on morphological criteria, it appears that chick limb mesenchymal cell osteogenesis in vitro is not associated with chondrogenesis but represents a separate route of phenotypic expression.
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  • 21
    ISSN: 1432-0827
    Keywords: Rat ; Calvarium ; Electron microscopy ; Preosteoclasts ; Osteoclasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary This is a study of the fine structure of cells of the 20-day fetal rat calvarium. Special attention is given to identifying and characterizing preosteoclasts. These cells are relatively common and located largely, but not exclusively, at the endocranial bone surface. The preosteoclasts are characterized by abundant mitochondria, an incomplete perinuclear Golgi apparatus, and variable-shaped dense granules. The dense granules are unique in appearance in that they contain an internal dense matrix surrounded by a clear halo. Most granules are circular in shape but some are elongate or tubular in form. Granules with identical appearance are observed in osteoclasts. The preosteoclasts are mononucleate, or occasionally binucleate. It is suggested that because preosteoclasts are morphologically distinctive and relatively abundant, it should be feasible to separate these cells from a heterogeneous cell isolate.
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  • 22
    ISSN: 1432-0827
    Keywords: Decalcification ; Electron microscopy ; Bone matrix ; Bone glycoproteins
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    Notes: Summary A solution of triethylammonium EDTA in 80% ethanol was evaluated as a demineralizing reagent for bone in comparison with aqueous solutions of EDTA. Biochemical analysis and acrylamide gel electrophoresis of extracts of finely powdered bovine bone showed that most of the macromolecular components of the organic matrix extractable in aqueous EDTA were retained when the triethylammonium EDTA reagent was used. Ultrastructural examination of chick tibias decalcified with the reagents showed a better preservation of cellular morphology, especially the membranous components, and more uniformly distributed ground substance, though slightly less in quantity, when the aqueous reagent was used. Use of the two reagents appears to be complementary, the alkylammonium reagent being more appropriate for use in studies of the organic matrix of bone, including immunohistochemical studies of bone glycoproteins. The aqueous reagent is more appropriate for use in studies of cellular ultrastructure.
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  • 23
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    Calcified tissue international 32 (1980), S. 229-236 
    ISSN: 1432-0827
    Keywords: Bone ; Ion influxes ; Calcium ; Phosphate ; Exchange
    Source: Springer Online Journal Archives 1860-2000
    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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  • 24
    ISSN: 1432-0827
    Keywords: Calcium ; Vitamin D deficiency ; 1,25(OH)2D3 ; Parathyroidectomy ; Parathyroid hormone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    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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  • 25
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    Calcified tissue international 4 (1969), S. 174-179 
    ISSN: 1432-0827
    Keywords: Calcium ; Strontium ; Bone ; Mineral
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé La proportion Sr/Ca dans l'os est à peu près la même que dans le sérum tandis que les poudres d'hydroxyapatite suspendues dans des solution physiologiques font une distinction contre le strontium. On trouva que la proportion Sr/Ca des fractions de différentes densités d'os compact de tibia de rat en forme de poudre augmentait avec augmentation de la densité jusqu'à une valeur un peu supérieure à celle observée dans le sérum. L'explication pourrait être qu'une barrière cellulaire ôte le calcium par préférence au strontium du fluide d'os ou bien qu'une accentuation précoce de la proportion Sr/Ca dans l'os se réfléchit dans les fractions de densité supérieure.
    Abstract: Zusammenfassung Das Verhältnis Sr/Ca im Knochen ist annähernd dasselbe wie es im Serum vorliegt, während in einer physiologischen Lösung von pulverisiertem Hydroxyapatit das Sr zugunsten des Ca benachteiligt wird. Es wurde festgestellt, daß das Sr/Ca-Verhältnis von Fraktionen verschiedener Dichte von pulverisierten Rattentibiae mit zunehmender Dichte anstieg, und zwar bis zu einem Wert, der etwas höher lag als der im Serum beobachtete. Dies ließe sich so erklären, daß eine Zellbarriere beim Entzug aus der Knochenflüssigkeit Ca dem Sr vorzieht; oder aber daß sich eine frühzeitige Betonung des für den Knochen typischen Sr/Ca-Verhältnisses in den Fraktionen höherer Dichte widerspiegelt.
    Notes: Abstract The Sr/Ca ration in bone is approximately the same as in serum, whereas hydroxyapatite powders suspended in physiological solutions discriminate against strontium. It was found that the Sr/Ca ratio of various density fractions of powdered compact bone of rat tibia increased with increasing density to a value slightly higher than that observed in the serum. The explanation could be that a cellular barrier removes calcium preferentially to strontium from the bone fluid, or that early accentuation of the bone Sr/Ca ration is reflected in the higher density fractions.
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    Calcified tissue international 4 (1969), S. 188-192 
    ISSN: 1432-0827
    Keywords: Serum ; Calcium ; Phosphate ; Calcitonin ; Thyroidectomy ; Metabolism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Après ablation des glandes thyroïdes de chiens, sans lèser les glandes parathyroïdiennes supérieurs, les taux en calcium et phosphate sériques et la gravité spécifique sont comparées avec des valeurs d'un groupe de chiens, chez lesquels une intervention simulée est réalisée. Immédiatement après thyroïdectomie, le taux de phosphate est plus élevé que le taux de calcium. Ce taux de phosphate se maintient plus longtemps à un niveau élevé que le taux de calcium, après thyroïdectomie. Le rapport entre l'augmentation du phosphate et le taux initial de phosphate sérique chez chaque animal indique des variations normales de la calcitonine d'un animal à l'autre.
    Abstract: Zusammenfassung Bei Hunden wurde die Thyreoidea entfernt, ohne die Parathyreoidea zu verletzen. Der Gehalt des Serums an Calcium und Phosphat sowie sein spezifisches Gewicht wurden mit den Werten verglichen, welche nach einer Scheinoperation bei einer Kontrollgruppe von Hunden bestimmt wurden. Unmittelbar nach der Thyreoidektomie war der Phosphatspiegel höher als jener des Calciums. Der Phosphatspiegel blieb nach der Thyreoidektomie länger hoch als der Calciumspiegel. Das Verhältnis zwischen dem Phosphatanstieg und dem Ausgangswert des Serumphosphates bei jedem Tier wies auf normale Variationen der Calcitonin-Konzentration bei verschiedenen Tieren hin.
    Notes: Abstract The thyroid glands were removed from dogs without interfering with the superior parathyroid glands. Levels of serum calcium, phosphate, and specific gravity were measured subsequently for 24 hours, and these values were compared with values obtained from a control group of dogs in which a mock operation was done. Immediately after thyroidectomy, the level of phosphate was greater than the calcium level. The phosphate level remained high longer after thyroidectomy than did the calcium level. The relationship between the increase in phosphate level and the initial serum phosphate value in each animal indicated normal variations in calcitonin levels in different animals.
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    Calcified tissue international 4 (1969), S. 245-256 
    ISSN: 1432-0827
    Keywords: Bone ; Densitometry ; X-ray ; Diet ; Calcium ; Osteoporosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé L'ingestion quotidienne de calcium of été déterminée en interrogeant 398 individus, âgés de 15 à 90 ans. Le rapport entre l'ingestion de calcium et la minéralisation vertébrale, mesurée par densitométrie radiographique quantitative, est fiable mais cependant significatif. Chez 53 personnes, atteintes d'ostéoporose, comparées à un nombre égal d'individus, d'âges similaires, les valeurs de la minéralisation vertébrale sont 60 pour cent plus faibles que celles du groupe témoin. L'ingestion totale moyenne de calcium est plus faible de 21 per cent chez les ostéoporotiques. Chez les sujets, à ingestion unique de calcium, les patients du groupe témoin ingèrent preque le double de calcium que le groupe des ostéoporotiques. Une diminution moyenne de l'ingestion calcique est notée avec l'âge. Il semble qu'il existe une diminution de l'absorption calcique avec l'âge, l'ostéoporose ou les deux, ainsi qu'un besoin plus grand en calcium chez les sujets âgés pour assurer l'équilibre en calcium. Indépendamment de l'homéostase calcique, un équilibre négatif en calcium augmente la résorption osseuse, et diminue l'ostéogenèse. De nombreux facteurs interviennent dans l'étiologie et la pathogénie de l'ostéoporose: les résultats de cette étude indique que la présence de calcium alimentaire est l'un de ces facteurs.
    Abstract: Zusammenfassung Die tägliche Calciumeinnahme wurde anhand einer Befragung von 398 Personen im Alter von 15–90 Jahren geschätzt. Die Korrelation der Calciumeinnahme und der Mineralisation der Wirbelsäule, wie sie röntgenologisch durch quantitative Messung der Dichte festgestellt wurde, war niedrig, aber durchwegs signifikant. Bei 53 Osteoporose-Patienten, die mit 53 Kontrollpersonen gleichen Alters verglichen wurden, waren die Mineralisationswerte der Wirbelsäule um 60% niedriger als jene der Kontrollgruppe, und die durchschnittlich geschätzte Gesamt-Calciumeinnahme der Osteoporotiker war um 21% tiefer. Von den Exploranden, die Angaben über eine gleichmäßige Calciumaufnahme machten, nahmen die Kontrollpersonen beinahe doppelt so viel Calcium ein als diejenigen mit Osteoporose. Im Durchschnitt wurde eine Verminderung der Calciumeinnahme mit fortschreitendem Alter festgestellt. Es ergibt sich deutlich, daß die Calciumabsorption mit dem Alter, bei Osteoporose oder in der Kombination dieser beiden Faktoren abnimmt, und daß eine größere Calciumeinnahme bei älteren Personen nötig wäre, um eine positive Calciumbilanz aufrechtzuerhalten. Ungeachtet der Komplexität der Calciumhomöostase, kann eine negative Calciumbilanz schließlich dazu führen, daß mehr Knochen resorbiert als gebildet wird. Demzufolge sollte bei erkanntem Bedürfnis eine adäquate Calciumeinnahme gesichert sein. Es ist eine Tatsache, daß viele Faktoren an der Ätiologie und Pathogenese der Osteoporose beteiligt sind; die Resultate dieses Berichtes unterstützen die Wahrscheinlichkeit, daß das Calciumangebot in der Nahrung einer dieser Faktoren ist.
    Notes: Abstract Lifetime daily calcium intake was estimated through interview of 398 individuals from 15 to 90 years of age. The correlation of calcium intake with vertebral mineralization as determined by quantitative radiographic densitometry was low but persistently significant. In 53 persons with osteoporosis matched by age with 53 individuals from the control group, vertebral mineralization values were 60% lower than those of the control group, and the mean estimated total calcium intake in osteoporotics was 21% lower. In those persons reporting a single lifetime calcium intake, the control patients ingested almost twice as much calcium as those with osteoporosis. A mean decrease in calcium intake with advancing years has been shown. Evidence points to a decrease in calcium absorption with age, osteoporosis, or both, as well as a greater need for calcium intake in the elderly to maintain a positive calcium balance. Regardless of the intricacies of calcium homeostasis, a negative calcium balance leads eventually to greater bone resorption than formation, hence the rationality of insuring an adequate calcium intake with recognized nutritional needs. Evidence suggests that many factors are involved in the etiology and pathogenesis of osteoporosis; the data in this report support the likelihood that availability of calcium in the diet is one of them.
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  • 28
    ISSN: 1432-0827
    Keywords: Calcitonin ; Parathyroid hormone ; Calcium ; Newborn ; Fracture
    Source: Springer Online Journal Archives 1860-2000
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    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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  • 29
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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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    Calcified tissue international 3 (1969), S. 100-102 
    ISSN: 1432-0827
    Keywords: Mitochondria ; Calcium ; Mineralisation
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    Calcified tissue international 2 (1968), S. 67-76 
    ISSN: 1432-0827
    Keywords: Calcergy ; Calciphylaxis ; Calcium ; Metals
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Chez le rat, des expériences ont montré que, sauf de rares exceptions, les calcifications produites au site d'injection d'acétate de plomb, de chlorure de cérium, chlorure de calcium ou de permanganate de potassium sont complètement prévenues ou fortement inhibées par l'application locale simultanée de provocateurs calciphylactiques, mais non par des agents non provocateurs. Malgré quelques exceptions (attribuées à des interrelations chimiques spécifiques entre certaines des substances utilisées), cette curieuse corrélation entre l'efficacité calciphylactique provocatrice et le pouvoir anticalcergique est, d'après le test χ2, hautement significative (P〈0,001).
    Abstract: Zusammenfassung Nach subcutaner Injektion von Bleiacetat, Ceriumchlorid, Calciumchlorid oder Kaliumpermanganat tritt an der Injektionsstelle eine starke Verkalkung auf. Bis auf sehr wenige Ausnahmen kann diese durch gleichzeitige lokale Anwendung von calciphylaktischen Provokatoren verhütet oder weitgehend unterdrückt werden. In dieser Beziehung sind nicht provolatorisch wirkende Stoffe inaktiv. Trotz der wenigen Ausnahmen (die wahrscheinlich auf spezifische, chemische Wechselwirkungen zwischen manchen der untersuchten Substanzen beruhen) ist dieser merkwürdige Zusammenhang zwischen calciphylaktisch provozierender und anticalcergischer Aktivität, nach dem χ2-Test berechnet, statistisch hochsignifikant (P〈0,001).
    Notes: Abstract Experiments on rats indicate that, with very few exceptions, the calcification which occurs at subcutaneous sites of treatment with lead acetate, cerium chloride, calcium chloride or potassium permanganate is completely blocked or severely inhibited by simultaneous topical application of calciphylactic challengers but not of non-challengers. Despite the few exceptions (which are thought to depend upon specific chemical interactions between some of the compounds tested) this singular correlation between calciphylactic challenging and anticalcergic potency, is highly significant by the Chi-square-test (P〈0.001).
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  • 32
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    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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  • 33
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    Keywords: Hydroxyapatite ; Dissolution ; Pyrophosphate, Diphosphonates ; Calcium ; Phosphate
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    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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    Calcified tissue international 31 (1980), S. 231-238 
    ISSN: 1432-0827
    Keywords: Magnesium ; Bone ; Calcium ; Parathyroid gland
    Source: Springer Online Journal Archives 1860-2000
    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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  • 36
    ISSN: 1432-0827
    Keywords: 1α-Hydroxy vitamin D3 ; 1,25-Dihydroxyvitamin D3 ; Calcium ; Transport ; Intestine
    Source: Springer Online Journal Archives 1860-2000
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    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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  • 37
    ISSN: 1432-0827
    Keywords: Perifusion ; Parathyroid hormone ; In vitro ; Calcium ; Beta regulation
    Source: Springer Online Journal Archives 1860-2000
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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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  • 38
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    Calcified tissue international 32 (1980), S. 29-44 
    ISSN: 1432-0827
    Keywords: Rabbit ; Dentin ; Calcium ; Sulfur ; Periodicity ; Circadian ; Ultradian
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    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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  • 39
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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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  • 40
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    Calcified tissue international 36 (1984), S. 596-603 
    ISSN: 1432-0827
    Keywords: Calcium ; Ameloblasts ; X-ray microanalysis ; Transport ; Frozen
    Source: Springer Online Journal Archives 1860-2000
    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 4 (1969), S. 180-184 
    ISSN: 1432-0827
    Keywords: Strontium ; Calcium ; Bone ; Shell
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Des poules ont été nourries avec une alimentation riche en strontium stable. Des concentrations de strontium de 3000 p.p. m. à 50000 p.p.m. montre une augmentation nette en strontium du tibia, alors que le contenu en calcium n'est pas modifié. Les concentrations en calcium sérique diminue, lorsque le strontium alimentaire augmente. Le calcium des coquilles d'oeufs diminue progressivement avec l'augmentation du strontium alimentaire, alors que le contenu en strontium des coquilles présente une augmentation correspondante. Des analyses de diffraction par rayons X des os et des coquilles ne permettent pas de déterminer sous quelle forme le strontium est déposé.
    Abstract: Zusammenfassung Die Resultate einer Verfütterung von hohen stabilen Strontiumdosen an Hühnern werden erläutert. Strontiumzusätze zur Nahrung in der Höhe von 3000–50000 p.p.m. führten zu einer signifikanten Zunahme des Strontiumgehaltes der Tibia und verursachten keine wesentlichen Änderungen des Calciumgehaltes. Die Calciumkonzentration im Plasma verminderte sich, wenn der Nahrung ansteigende Strontiummengen zugegeben wurden. Mit zunehmendem Strontiumzusatz zur Nahrung zeigte der Calciumgehalt der Eischale eine fortlaufende Abnahme, während sich der Strontiumgehalt entsprechend erhöhte. Durch Röntgendiffraktionsanalysen der stark Strontium-haltigen Knochen und Eischalen konnte nicht festgestellt werden, in welcher Form das Strontium abgelagert wurde.
    Notes: Abstract The results of feeding high dietary levels of stable strontium to hens are reported. Dietary levels of strontium from 3,000 p.p.m. to 50,000 p.p.m. showed a significant increase in strontium content of the tibia bone and essentially no change in the calcium content. Plasma calcium concentration was shown to decrease with increasing dietary strontium treatment. Egg shell calcium showed a progressive decrease with increasing dietary strontium treatment, whereas the strontium content has a corresponding increase. X-ray diffraction analyses of bones and shells containing large amounts of strontium were unsuccessful in evaluating the from in which strontium was deposited.
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    Calcified tissue international 4 (1969), S. 193-201 
    ISSN: 1432-0827
    Keywords: Calcium ; Carbonate ; Gland ; Ultrastructure ; Spherolith
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les cellules activement sécrétantes des glandes riches en calcium deLumbricus terrestris ont une forme en dôme. La partie basale de la cellule présente de nombreux prolongements ramifiés. Ces prolongements contiennent de nombreuses mitochondries, ainsi que des invaginations de la membrane formant des vésicules qui migrent dans diverses parties de la cellule. Le noyau irrégulier est en position centrale. La région supranucléaire contient surtout de l'ergastoplasme, plusieurs appareils de Golgi et de nombreuses granules provenant de l'appareil de Golgi. Ces granules augmentent de taille et sont rejetées dans le sinus de la glande, où elles se modifient et se calcifient pour donner des sphérolithes. Des radioautographics glandulaires, obtenues 1/2 heure après injection de Ca45, montrent une localisation isotopique, située presque exclusivement dans les cellules; 4–24 heures après, l'isotope se localise dans les sphérolithes de la cavité glandulaire. Ces cellules semblent assurer deux fonctions liées à la production de sphérolithes: 1. absorption et transport de calcium de la région basale vers la surface cellulaire et 2. formation d'une matrice protéique qui se calcifiera.
    Abstract: Zusammenfassung Die aktivsezernierenden Zellen der calcifizierenden Drüsen vonLumbricus terrestris sind annähernd kuppelförmig gestaltet. Der untere Teil dieser Zellen besteht aus unzähligen ineinandergreifenden Zellfortsätzen. Diese Fortsätze enthalten zahlreiche Mitochondrien und membranöse Einstülpungen, welche Bläschen bilden, die zu den verschiedenen Regionen der Zelle wandern. Der Zellkern ist unregelmäßig und zentral angeordnet. Die supranukleäre Region ist zu einem großen Teil mit grobem endoplasmatischem Reticulum und mehreren Golgi-Komplexen sowie zahlreichen Granula, die von den Golgi-Komplexen stammen, ausgefüllt. Diese Granula vergrößern sich und werden in den Drüseninus ausgestoßen, wo sie schließlich reorganisiert und mineralisiert werden, um Sphärolithen zu bilden. 1/2 Std nach Injektion von Ca45 wurde die Drüse entfernt; eine Radioautographie zeigte, daß das Isotop beinahe ausschließlich in den Zellen lokalisiert war; 4–24 Std später befand sich das Isotopin den Sphärolithen der Drüsenhöhlung. Diese Zellen sind anscheinend an zwei für die Bildung von mineralisierten Sphärolithen notwendigen Funktionen beteiligt: 1. an der Calcium-Absorption und deren Transport von der Basalregion bis zur Zelloberfläche; 2. an der Bildung einer Proteinmatrix, in welcher die Mineralisation stattfindet.
    Notes: Abstract The actively-secreting cells of the calciferous glands ofLumbricus terrestris are roughly dome shaped. The basal part of the cell consists of numerous interdigitating cell processes. These processes contain numerous mitochondria and membranous infoldings, forming vesicles which migrate to various regions of the cell. The nucleus is irregular and centrally located. The supranuclear region is occupied to a great extent by the rough endoplasmic reticulum, several Golgi complexes and numerous granules derived from the Golgi complexes. These granules enlarge and are extruded into the gland sinus, where they eventually become reorganized and mineralized to give rise to spheroliths. Radioautographs of glands removed 30 min following injection of45Ca showed uptake of the isotope located almost exclusively in the cells; 4–24 h later the isotope was located in the spheroliths in the gland cavity. These cells apparently perform two functions necessary for the production of mineralized spheroliths, 1, absorption and transport of calcium from the basal region to the cell surface and 2, the elaboration of a protein matrix in which mineralization occurs.
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    Calcified tissue international 4 (1969), S. 291-304 
    ISSN: 1432-0827
    Keywords: Bone ; Fluorescence ; Resorption ; Deposition ; Calcium ; Microradiography
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Divers agents chimiques, colorés, fluorescents et se localisant dans les os, à savoir la tétracycline, l'alazazine rouge S et le DCAF, ont été administré en série à des rats sevrés et on a mesuré le taux de la formation et de la résorption osseuse sur les coupes transversales du tiers supérieur de la diaphyse. Ici la formation osseuse périostale s'effectue progressivement avec peu de changement endostéal. Avec alimentation carnée, la croissance des rats est significativement restreinte pendant la première semaine, mais se rétablit ensuite. Bien qu'il y ait croissance des os, ceux-ci ne se minéralisent pas normalement et ils deviennent rapidement fragiles et amincis. La résorption osseuse est lente d'abord, puis s'accelère pendant 2–3 semaines pour atteindre un taux de 15μ par jour, après quoi elle se ralentit de nouveau. Bien que le taux de formation osseuse soit réduit, en comparaison avec celui des os normaux, la résorption s'effectue environ deux à trois fois plus rapidement que la croissance osseuse. Des études microradiographiques sur des rats à régime carné mais carencés en calcium ont permis la constatation suivante: tandis que la résorption s'effectue à la marge endostéale et que la formation osseuse a lieu sur l'aspect périostéal, la matière osseuse nouvellement formée est moins calcifiéc que chez les témoins.
    Abstract: Zusammenfassung Einige farbige, fluoreszierende, knochensuchende Chemikalien, z. B. Tetracyclin, Alazarin-Rot S und CDAF, wurden nacheinander an entwöhnten Ratten verabreicht, worauf man die Knochenbildungs- und Knochenresorptionswerte an hartgeschliffenen Schnitten des oberen Drittels der Diaphyse gemessen hat. Hier findet fortschreitend periostale Knochenbildung statt, mit geringer Veränderung des Endosteums. Bei Fleischdiät wird das Körperwachstum während der ersten Woche erheblich beschränkt; danach aber normalisiert es sich wieder. Obwohl die Knochen noch wachsen, zeigen sie keine normale Mineralisierung und werden schnell zerbrechlich und dünn. Die Knochenresorption ist anfangs langsam, dann beschleunigt sie sich während einer Zeitspanne von 2–3 Wochen bis auf 15 μ pro Tag, um sich dann wieder zu verlangsamen. Während die Knochenbildungsgeschwindigkeit relativ zum Normalwert heruntergesetzt wird, verläuft die Resorption ungefähr 2–3mal so schnell wie die Knochenbildung. Mikroradiographische Untersuchungen an mit Fleisch ernährten Ca-armen Ratten haben bestätigt, daß während die Resorption am Endosteumrande stattfindet und sich die Knochenbildung an der Periostenfläche fortsetzt, die neugebildete Knochensubstanz weniger kalzifiziert ist, als die der Kontrolltiere.
    Notes: Abstract Different coloured, fluorescent bone-seeking chemicals, viz., tetracycline, Alizarin Red S, and DCAF, have been administered sequentially to weanling rats and the rate of formation and resorption of bone measured from hard-ground cross sections of the upper third of the diaphysis of the femur. On a meat diet, bodily growth is significantly restricted for the first week and then recovery occurs. While bones grow they fail to mineralize normally and rapidly become fragile and rarefied. Resorption of bone is at first slow, then accelerates for a period of 2–3 weeks to about 15μ/day and then slows again. While the rate of bone formation is reduced relative to normal bone, resorption proceeds at approximately two to three times the rate of bone growth. Microradiographic studies confirm tht while resorption occurs on the endosteal margin and formation proceeds on the periosteal aspect of meat fed Ca-deficient rats, new bone is less calcified than that in control animals.
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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
    Source: Springer Online Journal Archives 1860-2000
    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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  • 47
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    Calcified tissue international 33 (1981), S. 119-124 
    ISSN: 1432-0827
    Keywords: Avian eggshell ; Microstructure ; Electron microscopy ; Electron diffraction ; Calcite growth
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    Topics: Biology , Medicine , Physics
    Notes: Summary The eggshell of the domestic fowl has been studied by transmission electron microscopy and diffraction. Thin sections of shell were prepared by chemical and ion-beam thinning techniques. Each calcite column of the palisade layer consisted of crystallites of diameter 20 to 30 µm with some tendency for crystallite alignment within a single column. Evidence indicates that there was no significant preferred orientation in the palisade layer as a whole. Only in the surface layer was any preferred orientation detected, and here {1014} planes tended to lie parallel to the surface. The results are compared with previously published data, and calcite nucleation and growth are discussed.
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    Calcified tissue international 2 (1968), S. 253-261 
    ISSN: 1432-0827
    Keywords: Calcium ; Ion ; Binding ; Chondroitin ; Cartilage
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    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé La liaison du calcium par le sulfate de chondroitin C a été étudiée en utilisant des méthodes biologiques et physicochimiques pour la détermination quantitative du calcium ionique. Les valeurs moyennes pour logK j, le logarithme de la constante de formation non corrigée d'un complexe calcium-sulfate de chondroitin C, sont 1,64 par la technique de cœur de grenouille, 1,55 par la méthode de murexide, 1,39 par l'ultrafiltration et 1,04 par l'électrode sélective du calcium. L'ordre de grandeur de ces valeurs suggèrent que le sulfate de chondroitin C a une capacité de liaison pour calcium relativement élevée, mais les valeurs calculées pour un paramètre d'échangeK p indiquent que le calcium a une affinité faible pour le polysaccharide.
    Abstract: Zusammenfassung Die Bindung von Calcium mit Chondroitinsulfat C wurde untersucht mittels biologischer und physikalisch-chemischer Methoden für die quantitative Bestimmung des ionischen Calciums. Im Durchschnitt ergab logK j, der Logarithmus der unkorrigierten Formationskonstante für einen Calciumchondroitinsulfatkomplex, 1,64 mit der Technik des Froschherzens, 1,55 mit der Murexidmethode, 1,39 mittels Ultrafiltration und 1,04 mit den Calciumselektiven Elektroden. Die Größe dieser Werte zeigt, daß Chondroitinsulfat eine relativ hohe Bindungskapazität für Calcium hat, die berechneten Werte für einen AustauschparameterK p jedoch, daß Calcium eine kleine Affinität für Polysaccharide hat.
    Notes: Abstract The binding of calcium by chondroitin sulfate C was studied by employing biological and physicochemical methods for the quantitative determination of ionic calcium. Mean values for logK j, the logarithm of the uncorrected formation constant for a calcium-chondroitin sulfate complex, were 1.64 with the frog heart technique, 1.55 with the murexide method, 1.39 with ultrafiltration and 1.04 with the calcium selective electrode. The magnitude of these values suggests that chondroitin sulfate has a relatively high binding capacity for calcium, but calculated values for an exchange parameterK p indicate that calcium has a low affinity for the polysaccharide.
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    Calcified tissue international 2 (1968), S. 301-313 
    ISSN: 1432-0827
    Keywords: Calcium ; Absorption ; Vitamin D
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    Calcified tissue international 2 (1968), S. 20-29 
    ISSN: 1432-0827
    Keywords: Bone Atrophy ; Bone Resorption ; Calcium ; Collagen ; Osteoporosis
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    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Atrophie osseuse d'immobilisation était induite par section de la plexus brachial et/ou désarticulation de la coude. Après 9 bis 12 semaines de désuétude huméri intactes furent examinés par rayons X et leur qualités physiques déterminés. Les humérientiers furent isolés, dégraissés, desséchés au poids constant, et leur teneur en cendre d'os et collagène déterminés. Huit des 10 huméri immobilisés monstrérent radiodensité diminué. La jambe inusitée monstra perte parallèle en poids sec et poids sans gras (−23.2%), en collagène (−25.3%), et en cendre d'os (−26.1%) en comparaison de la jambe normale. Les données monstrèrent que la portion plus grande du tissu osseux perdu en atrophie osseuse d'immobilisation est remplacée par de l'eau, du gras, et des autres matériels organiques inconnus plutôt que par tissu fibreux, et que collagène est perdu en proportion du minéral. La perte proportionellement plus grande du collagène et de la cendre d'os que du poids sec et poids san gras semble dû à une augmentation du matériel organique, non-collagéne, non-lipide, kprobablement protéine.
    Abstract: Zusammenfassung Immobilisations-Knochenatrophie wurde in 10 erwachsenen Hunden herbeigeführt durch Brachialplexusschnitt und/oder Ellenbogenexartikulation. Nach 9–12 Wochen der Immobilisation wurden die intakten Humeri unter Röntgenstrahlen untersucht und ihre physischen Eigenschaften bestimmt. Derintakte Humerus wurde isoliert, entfettet, zu konstantem Gewicht getrocknet und der Knochenaschengehalt und Kollagengehalt bestimmt. Acht der 10 experimentellen (immobilisierten) Humeri demonstrierte Beweis von verringerter Röntgendichte. Die immobilisierte Extremität zeigte ähnlichen Verlust in trockenem, fettfreiem Gewicht (−23,2%) in Kollagen (−25.3%) und Knochenasche (−26,1%) im Vergleich zur normalen Extremität. Die Date deuten an, daß der Hauptanteil des Verlustes des Knochengewebes (bei Immobilisations-Knochenatrophie) von Wasser, Fett und anderen unidentifizierten organischen Substanzen ersetzt wird, als von Faserngewebe und daß Kollagen und Knochenasche im gleichen Verhältnis verloren gehen. Der größere, proportionale Verlust an Kollagen und Knochenasche, eher als der Verlust an fettfreiem Gewicht, scheint in der Zunahme an nicht kollagener, nicht lipoider, organischer Substanz, vermutlich Protein, zu liegen.
    Notes: Abstract Disuse bone atrophy was induced in 10 adult dogs by means of brachial plexus section and/or elbow disarticulation. After 9 to 12 weeks of disuse intact humeri were examined by X-ray, and their physical properties determined. Thewhole humeri were isolated, defatted, dried to constant weight, and their mineral and collagen content determined. Eight out of 10 experimental (non-used) humeri demonstrated evidence of decreased radiodensity. The non-used limb demonstrated parallel loss in dry, fat-free weight (−23.2%), in collagen (−25.3%), and in mineral (−26.1%), as compared to the normal limb. The data indicated that the major portion of the lost bone tissue in disuse osteoporosis is replaced by water, fat, and other unidentified organic materials rather than fibrous tissue, and that collagen is lost in equal proportion to mineral. The proportionatelly greater loss of collagen and mineral than of dry, fat-free weight appears to be due to an increase of non-collagenous, non-lipid organic material, presumably protein.
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    Calcified tissue international 30 (1980), S. 27-34 
    ISSN: 1432-0827
    Keywords: Ultrastructure ; Calcium ; Cartilage ; Vesicles
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    Topics: Biology , Medicine , Physics
    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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  • 52
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    Calcified tissue international 34 (1982), S. 43-48 
    ISSN: 1432-0827
    Keywords: Calcium ; Phosphate ; Phosphatidylserine ; Transport ; Mineralization
    Source: Springer Online Journal Archives 1860-2000
    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
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    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 34 (1982), S. 121-124 
    ISSN: 1432-0827
    Keywords: Bone ; Calcium ; Metabolism
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    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 35 (1983), S. 107-110 
    ISSN: 1432-0827
    Keywords: Calcium ; Glucocorticoid ; Vitamin D ; Osteoporosis ; Rat
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    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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  • 59
    ISSN: 1432-0827
    Keywords: 31P NMR spectroscopy ; Phosphoprotein ; Dentin ; Calcium ; Inorganic orthophosphate ; Bovine
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    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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  • 60
    ISSN: 1432-2048
    Keywords: Calcium ; Cation exchange ; Lycopersicon esculentum ; Rhythm ; diurnal Transport (calcium) ; Zea mays
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    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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    Planta 158 (1983), S. 320-328 
    ISSN: 1432-2048
    Keywords: Calcium ; Egeria ; Fluorescent probe ; Ions, group II ; Symplast
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    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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  • 62
    ISSN: 1432-2048
    Keywords: Bryophyta ; Bud formation (moss) ; Calcium ; Chlorotetracycline ; Cytokinin ; Funaria
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    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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    Planta 152 (1981), S. 450-456 
    ISSN: 1432-2048
    Keywords: Calcium ; Cell elongation ; Gibberellin ; Lactuca ; pH and growth
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    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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    European biophysics journal 7 (1981), S. 209-212 
    ISSN: 1432-1017
    Keywords: Photosynthetic bacteria ; Electron microscopy ; Planar lattices
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    Topics: Biology , Physics
    Notes: Abstract The thylakoids of Rhodopseudomonas viridis have been studied by freeze-fracturing whole cells. Depending on growth conditions and treatment before freezing, three different types of particle arrangements in the photosynthetic membrane are reported: a random arrangement, an isometric (quadratic) lattice arrangement with a lattice constant of 12.5 ± 0.8 nm, and a hexagonal lattice arrangement with a lattice constant of 12.5 ± 0.8 nm.
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    European biophysics journal 9 (1982), S. 131-136 
    ISSN: 1432-1017
    Keywords: Chemotaxis ; Calcium ; Oscillation ; Dictyostelium ; Ionophore
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    Topics: Biology , Physics
    Notes: Abstract Migration of differentiated cells to a capillary containing cyclic AMP was enhanced in the presence of 1 mM CaCl2 and was virtually absent in the presence of 1 mM EGTA. Furthermore, the cells contracted and extended pseudopods to a capillary filled with the calcium ionophore A 23187. At short distances, migration to the tip of the capillary was observed. The ionophore also induced transient decreases of the optical density of suspended cells indicating changes of cell shape. These findings support the hypothesis that cyclic AMP-binding to cell surface receptors causes a local influx of calcium ions. These in turn lead to an increase of the cytosolic calcium concentration and subsequently to an activation of cell migration. Perturbing pulses of the ionophore induced permanent phase shifts of free-running light scattering oscillations. This result indicates that cytosolic calcium is an intrinsic component of the oscillatory system.
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  • 66
    ISSN: 1432-0983
    Keywords: Allomyces macrogynus ; Mitochondrial DNA ; Electron microscopy ; Restriction enzyme map
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    Topics: Biology
    Notes: Summary The mitochondrial (mt) DNA of the aquatic phycomycete Allomyces macrogynus is a circular molecule with a size of 56.1 kbp. The cleavage sites for the restriction enzymes SalI and PvuI were mapped by comparing the partial denaturation patterns of isolated restriction fragments with the pattern of the intact circle. The genes coding for the small and large ribosomal RNA (rRNA) were located on the restriction map by heteroduplex and R-loop analysis. The gene coding for the large rRNA contains an intervening sequence, app. 0.7 kbp in size, near the 3′-end of the gene. The two rRNA genes are encoded on the same strand of the mtDNA and separated by a region of 17–18 kbp. This rRNA gene organization is similar to that found with members of the Ascomycetes.
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  • 67
    ISSN: 1432-072X
    Keywords: Allomyces ; Zoospores ; Cell wall ; Chitin ; Gamma particle ; Encystment ; Electron microscopy ; Calcofluor
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    Topics: Biology
    Notes: Abstract Structural changes during cell wall formation by populations of semisynchronously germinating zoospores were studied in the water mold Allomyces macrogynus. Fluorescence microscopy using Calcofluor white ST (which binds to β-1,4-linked glycans) demonstrated that Calcofluor-specific material was deposited around most cells between 2–10 min after the induction of encystment (beginning when a wall-less zoospore retracts its flagellum and rounds up). During the first 15 min of encystment there was a progressive increase in fluorescence intensity. Ultrastructural analysis of encysting cells showed that within 2–10 min after the induction of encystment small vesicles 35–70 nm diameter were present near the spore surface, and some were in the process of fusing with the plasma membrane. The fusion of vesicles with the zoospore membrane was concomitant with the appearance of electron-opaque fibrillar material outside the plasma membrane. Vesicles similar to those near the spore surface were found within the gamma (γ) particles of encysting cells. These particles had a crystalline inclusion within the electron-opaque matrix. During the period of initial cyst cell wall formation numerous vesicles appeared to arise at the crystal-matrix interface. Approximately 15–20 min was required for the cell wall to be formed. We suggest that the initial response of the zoospore to induction of encystment is the formation of a cell wall mediated by the fusion of cytoplasmic vesicles with the plasma membrane.
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    Archives of microbiology 130 (1981), S. 125-128 
    ISSN: 1432-072X
    Keywords: Rhodospirillum rubrum ; Chromatophores ; Reaction centers ; Liposomes ; Electron microscopy
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    Topics: Biology
    Notes: Abstract In freeze-fractures of chromatophores of Rhodospirillum rubrum the reaction centers are seen as hexagonal arranged particles of 13 nm diameter with a density of around 5,500 particles per μm2. Similar regions on the cytoplasmic membrane suggest that these parts are the prospective invagination sites. Isolated reaction centers are easily incorporated into liposomes. In freeze fractures of liposomes particles similar in shape and size, although less dense as in chromatophores are observed. In negative staining much smaller units of only 5 nm in diameter are found indicating that reaction centers occur in the membrane as tri- or tetramers. There is a strong correlation between particle density in chromatophores and titratable reaction centers remaining in these membranes after extraction of reaction centers by detergents; both values are in good agreement with the yield of reaction centers at a given detergent concentration.
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    Archives of microbiology 126 (1980), S. 277-283 
    ISSN: 1432-072X
    Keywords: Bacteriophage ; Myxococcus ; λ ; Superooiled DNA ; Cross-linking ; Electron microscopy
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    Topics: Biology
    Notes: Abstract DNA was partially released from the heads of myxococcus phages and also coliphage λ and examined by electron microscopy by a modification of the Kleinschmidt technique, in which water was used as hypophase. DNA emerged from the heads in patterns suggestive of newly relaxed supercoils. The unreleased DNA appeared to occupy discrete regions in the head. Some closed circles were released from λ heads. When NaCl solution was used as hypophase, the DNA was observed either released from the tail or from the head, in the latter case, supercoiled regions were observed. When NH4OAc solution was used as hypophase, tightly wound structures were released from λ heads; these fields also contained supercoiled circles. The presence of constrained supercoiled domains in newly released phage DNA was confirmed by observing the effects of ethidium bromide on its conformation. Treatment of phage with nitrogen mustard, a bifunctional alkylating agent, preserved supercoiled domains, even when the phage were lysed over water as hypophase. Further experiments suggested that phage inactivation by nitrogen mustard is largely due to restraint of the supercoiled, native, tertiary structure and that DNA-protein cross-linking may be involved in this reaction. The implications of these findings for the conformation of phage DNA in vivo are discussed and a new model for the winding of DNA in phage heads is proposed.
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    Archives of microbiology 129 (1981), S. 129-134 
    ISSN: 1432-072X
    Keywords: Cockroach ; Hindgut ; Distribution ; Microbial morphotypes ; Transmission ; Electron microscopy ; Statistical analysis ; Eublaberus posticus
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    Topics: Biology
    Notes: Abstract The relative numbers of fourteen microbial morphotypes in transmission electron micrographs of the hindgut of a cockroach, Eublaberus posticus, were counted and their distribution was analyzed statistically. The microbiota of three wall-associated regions (the anterior paunch, the posterior paunch, and the black band region) was clearly different from that of the gut lumen. The three wall fractions were also significantly different from each other. Only one of the fourteen types, prosthecate bacteria, appeared to be distributed randomly in the four fractions. The five main wall-associated morphotypes individually constituted up to 41% of the microbes in some micrographs. They included one type with the characteristic morphology of Methanospirillum. Six morphotypes rarely made up over 2% of the population, but were consistently present. The numbers of the remaining three morphotypes were quite variable between micrographs and between individual insects, but when present often made up 5–10% of the population.
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  • 71
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    Keywords: Biological control ; Cyanobacteria ; Electron microscopy ; Entrapment ; lysis ; Myxococcus ; Phormidium ; Spherule
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    Notes: Abstract A Myxococcus xanthus isolate from a farm drainage ditch, designated strain PCO2, is capable of rapidly inducing lysis of both agar and liquid-grown cultures of the cyanobacterium, Phormidium luridum, var. olivacea. Microscopic studies of the predator-prey interaction demonstrate that lysis of the cyanobacterium occurs within clumps and spherules formed by the cells of M. xanthus PCO2. In the earliest stage, one sees the formation of irregular microclumps of bacteria and cyanobacterial filaments. As these clumps mature, colonies 1 to 6 mm in diameter develops. The center of these densely green colonies contains cyanohacteria in various stages of degradation, while the periphery is almost exclusively a tightly woven mass of myxobacterial cells. Electron microscopy shows that long extrusions from the outer membrane of the M. xanthus PCO2 cells are involved in the formation both of initial clumps and of mature colonial spherules. These extrusions appear to efficiently entangle the cyanobacterial filaments in the culture environment. Predator-to-prey ratios of 1/10, 1/100 and 1/1,000 have resulted in cyanobacterial lysis. Because the entrapment and lysis of P. luridum filaments by M. xanthus PCO2 appears to be independent of any other heterotrophic nutritional requirement, as well as of environmental agitation, this system has potential as a biological control technique for undesirable aquatic cyanobacteria.
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  • 72
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    Archives of microbiology 138 (1984), S. 273-277 
    ISSN: 1432-072X
    Keywords: Cyanobacteria ; Secondary metabolite ; Allelopathy ; Photosynthesis ; Electron transport ; Thylakoids ; Herbicides ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cyanobacterin is a secondary metabolite produced by the cyanobacterium, Scytonema hofmanni. Highly purified cyanobacterin was found to inhibit the growth of many cyanobacteria at a minimum effective dose of 2 μg/ml (4.6 μM). The antibiotic had no effect on eubacteria including the photosynthetic Rhodospirillum rubrum. The site of action of cyanobacterin was further investigated in the unicellular cyanobacterium, Synechococcus sp. Electron micrographs of antibiotic-treated Synechococcus cells indicated that cyanobacterin affects thylakoid membrane structure. The antibiotic also inhibited light-dependent oxygen evolution in Synechococcus cells and in spheroplasts. These data support our conclusion that cyanobacterin specifically inhibits photosynthetic electron transport. This activity is similar to herbicides such as 3-(3,4-dichlorophenyl)-1,1-dimethyl urea (DCMU). The anhydro analog of cyanobacterin had no biological activity.
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  • 73
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    Archives of microbiology 130 (1981), S. 339-343 
    ISSN: 1432-072X
    Keywords: Leptospira ; Halophilic ; Electron microscopy ; X-ray analysis ; Inclusions ; Cytoplasmic tubules
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    Topics: Biology
    Notes: Abstract The morphology of cells of strain Muggia, a slightly halophilic leptospire, was examined by the negative staining technique. The ultrastructure of the cells was rather similar to that of cells of Leptonema illini, i. e. the cells possessed cytoplasmic tubules. The basal complex of their flagella, however, was similar to the corresponding part of flagella on Gramnegative bacteria. The interior of the cells was densely packed with inclusions, except for the two outermost wavelengths at each end where these inclusions were absent. X-ray microanalysis showed that the inclusions contained sodium and chlorine as their main constituents. The inclusions disappeared upon storage of the cultures at room temperature.
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  • 74
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    Archives of microbiology 132 (1982), S. 10-13 
    ISSN: 1432-072X
    Keywords: Calcofluor White ; Cell wall structure ; Chlorella ; Electron microscopy ; Protoplast ; Ruthenium Red
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Among 12 strains ofChlorella ellipsoidea, C. vulgaris, andC. saccharophila tested, 4 strains (1,C. ellpsoidea; 2,C. vulgaris; 1,C. saccharophila) formed osmotically labile protoplasts after treatment with mixtures of polysaccharide degrading enzymes. The relationship between enzymatical digestibility and structure or composition ofChlorella cell walls were studied by electron microscopy and staining techniques with some specific dyes. The cell wall structures of the 12Chlorella strains were grouped into three types: (1) with a trilaminar outer layer, (2) with a thin outer monolayer, and (3) without an outer layer. Protoplasts were formed only from the strains with a cell wall of Type 2. In the strains with a cell wall of Type 1, the outer layer protected the inner major microfibrillar layer against enzymatic digestion. The cell wall of Type 3 was totally resistant to the enzymes; the chemical composition of the cell wall would be somewhat different from that of other types.
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  • 75
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    Archives of microbiology 135 (1983), S. 169-175 
    ISSN: 1432-072X
    Keywords: Photosynthetic membranes ; Electron microscopy ; Image processing ; Ectothiorhodospira halochloris ; Ectothiorhodospira abdelmalekii ; Rhodopseudomonas viridis ; Rhodopseudomonas sulfoviridis ; Thiocapsa pfennigii
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract The photosynthetic membranes of the five bchl b-containing bacteria Ectothiorhodospira halochloris, E. abdelmalekii, Rhodopseudomonas viridis, R. sulfoviridis and Thiocapsa pfennigii have been investigated by electron microscopy and digital image analysis. All five species have the photosynthetic complexes hexagonally arrayed in the membrane with lattice spacings close to 13 nm, except for R. sulfoviridis and T. pfennigii which display somewhat smaller (∼12.5 nm) lattice spacings. Correlation averaging which imposes less stringent requirements on the lattice perfection than conventional Fourier filtration techniques has been employed to elucidate the structure of the photosynthetic complexes. Their basic organization, i.e. a ring, probably containing the light-harvesting (LH) polypeptides, surrounding a core (the “reaction centre”) appears to be almost identical for all species under scrutiny. Despite a resolution of ∼1.6 nm, however, little further significant substructure can be deduced from the averages; possible reasons for the “blurred” appearance of the LH-ring and absence of any subdivision in the reaction centre are discussed along with strategies aimed at obtaining a more detailed model of the molecular architecture of the photosynthetic membranes.
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  • 76
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    Keywords: Phototaxis ; Motility ; Flagellate ; Chlamydomonas ; Sodium azide ; Calcium ; A 23 187
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    Topics: Biology
    Notes: Abstract A fully automated and computerized method for simultaneous measurements of motility and phototaxis of unicellular flagellates is described. Both systems are directly coupled with a homocontinuous culture. The motility measuring apparatus is equipped with a video camera and recorder for simultaneous single cell behaviour studies. First results of studies on the effects of the phototaxis inhibitor sodium azide and the Ca2+ conducting ionophore A23 187 on motility and phototaxis of Chlamydomonas are reported and correlated with video observations. These results demonstrate that the described systems give informations of whether phototaxis or motility or both are inhibited by chemicals.
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  • 77
    ISSN: 1432-072X
    Keywords: Immunoferritin labeling ; Electron microscopy ; Membrane vesicles ; Nitrate reductase ; Bacillus licheniformis ; Klebsiella aerogenes
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    Topics: Biology
    Notes: Abstract The indirect immunoferritin labeling method was used to localize the membrane-bound respiratory nitrate reductase in membrane vesicles and protoplasts or spheroplasts of Bacillus licheniformis and Klebsiella aerogenes, respectively. For a comparison of the labeling of the various vesicle preparations, which differed not only in size but also in the percentage of inside-out orientation, a quantification of the results was needed to circumvent the problem of non-specifically bound ferritin. From the results the sidedness of the nitrate reductase in the cytoplasmic membrane of the abovementioned bacteria was determined as being cytoplasmic in B. licheniformis and as transmembranous in K. aerogenes.
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  • 78
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    Archives of microbiology 128 (1980), S. 12-18 
    ISSN: 1432-072X
    Keywords: Moraxella glucidolytica ; Electron microscopy ; Lipopolysaccharide
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    Topics: Biology
    Notes: Abstract A cellular (LPS I) and extracellular (LPS II) lipopolysaccharide were isolated from Moraxella glucidolytica cells grown on ethanol and from the culture fluid, respectively. Both LPS were toxic when injected to mice and chick embryos. These LPS contained glucose, galactose, glucosamine, galactosamine, 2-keto-3-deoxyoctonate and lipids. By permethylation studies, glucose was found to be linked (1→6) and (1→3) in LPS I and only (1→6) in LPS II. Galactose was the terminal non-reducing sugar. Branching occurred at positions 3 and 4 of galactose residues. LPS I was rich in α- and β-hydroxylauric and α-hydroxymyristic acids and LPS II contained mainly stearic and α-hydroxymyristic acids. LPS I was detoxified by mild acid and alkaline treatments. It was also dissociated by sodium deoxycholate and chromatographed on Sephadex G-75. The main fraction was reassociated by removing the surfactant by dialysis. The morphology of LPS I and LPS II was examined by electron microscopy. LPS I (original and reassociated fractions) consisted exclusively of ribbons while LPS II contained ribbons and vesicles.
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  • 79
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    Archives of microbiology 138 (1984), S. 140-152 
    ISSN: 1432-072X
    Keywords: Caulobacter ; Prostheca development ; Stalk function ; Calcium ; Phosphate
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    Topics: Biology
    Notes: Abstract Calcium was found to stimulate stalk development in Caulobacter crescentus and to relieve the inhibition of development long known to be caused by phosphate. This suggested that phosphate inhibition could be attributed to its interaction with Ca2+, thereby depriving the cells of a factor that promoted development. Calcium was also found to promote phosphate acquisition by the cells, observed as acceleration of growth at extremes of phosphate concentration, as promotion of carbon-source utilization rather than storage, and as support for phosphate-dependent resistance to arsenate inhibition of growth. Cytological studies of dividing cells revealed that stalked siblings had greater access to exogenous phosphate for use in growth or for storage as polyphosphate, and that access of non-stalked sibling to phosphate was dependent on the length of the stalk of the dividing cell. It was concluded that the physiologic role of the stalk is enhancement of phosphate acquisition. The stimulatory role of calcium in this process was attributed to its support of stalk development and to its stabilization of internal membrane/cell envelope association within the cell-stalk juncture.
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  • 80
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    Keywords: Ribulose bisphosphate carboxylase ; Quaternary structure ; Molecular weight ; Electron microscopy ; Cyanobacteria ; Synechococcus
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    Notes: Abstract Ribulose bisphosphate (RuP2) carboxylase from the marme cyanobacterium, Synechococcus sp., comprised both large (57,000 dalton) and small (12,000 dalton) subunits. The undissociated, purified enzyme was considerably smaller than the spinach enzyme when compared by pore-gradient electrophoresis, gel filtration and density-gradient centrifugation. This suggested that the cyanobacterial enzyme might have a hexameric (L6S6) subunit structure, unlike the enzymes from spinach and many other organisms which are octamers (L8S8). However, the molecular weight of the Synechococcus enzyme was measured by equilibrium sedimentation and found to be 530,000, which is within the range observed for L8S8-type enzymes. Furthermore, electron microscopic studies of negatively stained preparations of both the native enzyme, and a preparation depleted of 87% of its small subunits by repeated mild-acid precipitation, revealed four-fold symmetry characteristic of an octameric, cubical structure. Synechococcus RuP2 carboxylase therefore must be an L8S8 octamer and its anomalous pore-penetration behaviour may be due to an asymmetric shape. Some support for the latter possibility was provided by electron miscoscopic observations of two different types of images which may be different views of the molecule in two planes.
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  • 81
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    Archives of microbiology 140 (1984), S. 265-270 
    ISSN: 1432-072X
    Keywords: EcoRI ; EcoRI-DNA complexes ; EcoRI* activity ; Recognition sites ; Frequency of binding ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Electron microscopy of negatively stained isolated restriction enzyme EcoRI revealed particle projections with triangular or square outlines, indicating that the enzyme, in its tetrameric state, is tetrahedron-like. The two dimers making up the tetramer appear to be arranged in two planes orthogonal to each other. Complexes formed by EcoRI with the plasmids pBR322 or pGW10 were investigated by electron microscopic spreading techniques. In the presence of Mg2+, EcoRI was bound to the DNA molecules to form pearl necklace-like aggregates. The number of bound EcoRI particles was much higher as the sum of EcoRI-and 5′..AATT..3′ sites (with exceptions, the 5′..AATT..3′ sites may function as one type of EcoRI* sites) along the DNAs, indicating unspecific binding. In the absence of Mg2+, EcoRI was bound to the DNA only at the recognition site for EcoRI and the sites where the tetranucleotide sequence 5′..AATT..3′ was present. A direct correlation of the local concentrations of the bases A and T within the flanking sequences of the binding sites with the frequency of EcoRI to the DNA was observed. Dimers and tetramers of the enzyme was found to bind to the DNA. Tetramers occasionally exhibited two binding sites for DNA as indicated by the observation of DNA loops originating at the sites of bound tetrameric EcoRI particles.
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  • 82
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    Archives of microbiology 126 (1980), S. 87-95 
    ISSN: 1432-072X
    Keywords: Ascodichaena ; Beech bark ; Electron microscopy ; Host-fungus relationship
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    Topics: Biology
    Notes: Abstract Ascodichaena rugosa Butin is a corkinhabiting fungus, found frequently on the bark of Fagus sylvatica L. The hyphae of the fungus are distributed solely in the phellem cells, stopping their growth in the last-formed cork cell layer. The cell to cell invasion is effected by penetration hyphae, causing no extensive dissolution of the cork wall. Electron microscopical observations revealed fine structural details of the fruit bodies and of the intracellular hyphae. Of special interest were the finger-like hyaline hyphae in the last-formed layer of cork cells, which are interpreted as haustoria on the basis of the fine structure both of hyphae and host cells. This situation is considered as reflecting a parasitic relationship of Ascodichaena to beech bark. The activity of the fungus led also to the increased production of cork cells, perhaps related to the nutrient supply of the fungus.
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  • 83
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    Archives of microbiology 133 (1982), S. 97-99 
    ISSN: 1432-072X
    Keywords: Cyanobacteria ; Thylakoid centers ; Photosynthetic membranes/thylakoids ; Membranes ; Membrane biogenesis ; Electron microscopy
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    Notes: Abstract An ultrastructural study of four cyanobacteria (Anabaena cylindrica, Dermocarpa violaceae, Gleocapsa alpicola, Pleurocapsa minor) indicates the presence of previously undescribed thylakoid centers from which photosynthetic membranes (thylakoids) radiate. These peripherally located thylakoid centers are cylinders 30 nm wide by 320 nm long, consisting of globular subunits oriented in nonparallel stacked arrays. Thylakoids are attached to the outer surface of the cylinder along its longitudinal axis. Thylakoid centers appear to be functionally significant due to their structure, location and thylakoid association.
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  • 84
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    Archives of microbiology 135 (1983), S. 25-29 
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    Keywords: Electron microscopy ; Euglena mutabilis ; Flagellate ; Photomovement ; Photoreceptor ; Phototaxis ; Single-cell analysis ; Videomicroscopy
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    Topics: Biology
    Notes: Abstract Due to the lack of an emergent flagellum the green flagellate Euglena mutabilis is restricted to gliding motility. During forward movement, the organisms orient positive phototactically in the presence of a suitable light stimulus. The cell contains both a stigma and a paraflagellar body which differ in shape and size from the organelles found in E. gracilis. The degree of orientation in white light follows an optimum curve with a maximum at about 100 lx. The spectral sensitivity shows a number of prominent peaks in the blue and green regions and extends well into the red region of the visible spectrum. Since the cell does not rotate during locomotion a periodic shading mechanism cannot account for phototactic orientation. Thus, phototaxis in the related species, E. gracilis and E. mutabilis differ in their photoreceptor molecules, their sensory transduction chains and their strategies of light direction detection.
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  • 85
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    Archives of microbiology 139 (1984), S. 102-104 
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    Keywords: Fungus ; Zoospore ; Blastocladiella ; Allomyces ; Gamma ; Particle ; Phosphate ; Calcium
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    Notes: Abstract The elemental composition of gamma particles in zoospores ofAllomyces macrogynus andBlastocladiella emersonii was determined by use of energy dispersive X-ray analysis of glutaraldehyde fixed, thin section zoospores. Isolated preparations of gamma particles were also examined. Gamma particles contained no detectable elements. Similar sized, globular, electron dense cytoplasmic inclusions contained phosphorus and calcium. We suggest that previous studies assigning calcium and phosphorus to gamma particles may have been based on confusion of these two types of cytoplasmic inclusions.
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  • 86
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    Archives of microbiology 131 (1982), S. 116-123 
    ISSN: 1432-072X
    Keywords: Cell wall ; Wall degradation ; Lysozyme ; Autolysines ; Electron microscopy ; Staphylococcus aureus
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    Notes: Abstract In contrast to former findings lysozyme was able to attack the cell walls ofStaphylococcus aureus under acid conditions. However, experiments with14C-labelled cell walls and ribonuclease indicated that, under these conditions, lysozyme acted less as an muralytic enzyme but more as an activator of pre-existing autolytic wall enzymes. Electron microscopic studies showed that under these acid conditions the cell walls were degraded by a new mechanism (i.e. “attack from the inside”). This attack on the cell wall started asymmetrically within the region of the cross wall and induced the formation of periodically arranged lytic sites between the cytoplasmic membrane and the cell wall proper. Subsequently, a gap between the cell wall and the cytoplasmic membrane resulted and large cell wall segments became detached and suspended in the medium. The sequence of lytic events corresponded to processes known to take place during wall regeneration and wall formation. In the final stage of lysozyme action at pH 5 no cell debris but “stabilized protoplasts” were to be seen without detectable alterations of the primary shape of the cells. At the same time long extended ribbon-like structures appeared outside the bacteria. The origin as well as the chemical nature of this material is discussed. Furthermore, immunological implications are considered.
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  • 87
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    Planta 160 (1984), S. 12-20 
    ISSN: 1432-2048
    Keywords: Calcium ; Calmodulin ; Germination (spore) ; Onoclea ; Phytochrome and Ca2+ ; Pteridophyta
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    Notes: Abstract Phytochrome is confirmed to be the photoreceptor pigment in the germination response of Onoclea sensibilis L. by demonstrating red-far-red (R-FR) photoreversibility. External Ca2+ is required for this response with a threshold at a submicromolar concentration. Ethylene glycol-bis(β-amino-ethyl ether)-N,N,N′,N′-tetraacetic acid, La3+ and Co2+ reversibly inhibit germination. Lanthanum only inhibits germination when applied before or during irradiation, indicating that the external Ca2+ requirement is transient, although in the absence of Ca2+ the R-stimulated system remains maximally poised to accept the ion for over 4 h after irradiation. The ability to respond to Ca2+ 4.1 h after R-irradiation is not reversed by FR-irradiation, indicating that Ca2+ transport has been uncoupled from phytochrome. Barium and Sr2+, but not Mg2+ can substitute for Ca2+. Artificially increasing the concentration of intracellular free Ca2+ with the ionophore A 23187 stimulates germination in the dark. The Ca2+-calmodulin antagonists, trifluoperizine and chlorpromazine, reversibly inhibit germination. Calcium is required in phytochrome-mediated fern spore germination; it may be acting as a second messenger.
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  • 88
    ISSN: 1432-0878
    Keywords: Ionophore A-23187 ; Calcium ; Exocrine pancreas ; Secretion ; Zymogen granule
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    Topics: Biology , Medicine
    Notes: Summary Rat-pancreas lobules were incubated with the ionophore A-23187 in the presence of Ca2+. After 90 min, some of the acini were partially or almost completely depleted of their zymogen granules while others had the appearance of resting acini. With few exceptions, the cells of a given acinus were degranulated to a comparable level. Slight dispersion of the zymogen granules was noticed in cells incubated in a Ca2+-free medium containing EGTA with or without A-23187. In the presence of Ca2+ the secretory response obtained with the ionophore was comparable to that observed with 10-5M urecholine. The results obtained provide cytological evidence that the secretory response is only partially determined at the membrane-receptor level and that other mechanisms intervene between cytosol Ca2+ increase and exocytosis.
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  • 89
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    Cell & tissue research 237 (1984), S. 219-226 
    ISSN: 1432-0878
    Keywords: Spermatogonia ; Spermatocytes ; Carbohydrates ; Guppy ; Electron microscopy
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    Topics: Biology , Medicine
    Notes: Summary The structure of guppy (Poecilia reticulata) spermatogonia and spermatocytes has been studied using electron microscopy. The spermatogonia, situated at the apex of the seminiferous tubule, are almost all surrounded by a network of Sertoli cells; they have very diffuse chromatin and one or two large nucleoli. The cytoplasm contains relatively few organelles, although annulate lamellae are found. The mitochondria have few cristae and are concentrated at one pole of the cell; they are sometimes found with intermitochondrial cement. These spermatogonia are separated from each other, having no intercellular bridges or inclusion in Sertoli cells, and are relatively undifferentiated; they correspond to stem cells. The spermatogonia beneath the apex are organized into cysts. First-generation spermatogonia are more dense and heterogeneous, their nuclei becoming smaller and their chromatin becoming denser during successive generations. In spermatocytes, the synaptinemal complex exists as a modified form until metaphase. The concentration of organelles in the cytoplasm increases and the organelles become more diversified as spermatogenesis progresses. Many cytoplasmic bridges are observed (several per cell), indicating that the cells remain in contact after several divisions. These changes in germ cell structure have been related to some of the characteristic features of spermatogenesis in guppy, e.g. the large number of spermatogonial generations and the complexity of spermiogenesis.
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    Cell & tissue research 238 (1984), S. 413-416 
    ISSN: 1432-0878
    Keywords: Peroxisomes ; DAB-cytochemistry ; Electron microscopy ; Liver, amphibian ; Gymnophiona ; Ichthyophis glutinosus
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    Topics: Biology , Medicine
    Notes: Summary The ultrastructure of hepatic peroxisomes was investigated in Ichthyophis glutinosus (Amphibia: Gymnophiona), employing perfusion fixation and the diaminobenzidine (DAB) technique for the visualization of catalase. The majority of peroxisomes is circular or rod-shaped, although elongated particles occasionally occur. They contain a finely granular matrix, lightly stained after the DAB procedure. Their mean diameter is approximately 0.25 μm. Serial sections reveal that the circular and rod-shaped peroxisomal profiles are cross and oblique sections of highly tortuous, tubular organelles exceeding 2 μm in length. In addition to tubular profiles, elongated, rectangular particles, as well as straight dumbbell-shaped organelles with distinct marginal plates are observed. They range from 900 to 1650 nm in length (mean = 1200 nm). In the flattened, thin central portion of the dumbbell-shaped particle, the peroxisomal membranes form a cisterna enclosing one or two uniformly thick marginal plates, which display a definite substructure with a periodicity of 10 nm. These findings indicate that peroxisomes in the liver of Ichthyophis exhibit a complex organization. It is suggested that the organelles undergo a specific differentiation process, morphologically characterized by the formation of enlarged segments of unusual shape.
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    Cell & tissue research 101 (1969), S. 72-87 
    ISSN: 1432-0878
    Keywords: Cell differentiation ; Keratin ; Electron microscopy ; Reptiles ; Skin
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    Topics: Biology , Medicine
    Notes: Summary The epidermis of Anolis carolinensis is renewed periodically by molting. Prior to the molt the distinct layers of the epidermis, namely, the Oberhäutchen, β, mesos, and α layers, are formed in sequence from a morphologically homogenous population of basal cells. The Oberhäutchen, the first cell layer to form, has spinules on the surface which interdigitate with the overlying cells of the clear layer. The cells of the Oberhäutchen develop 80 Å filaments similar to those in the cells of the α layer. Beneath the Oberhäutchen is the β layer, the cells of which develop membrane-bounded packets containing a homogenous material during the early stages of differentiation. Later 100–500 Å thick fibrils are formed in the membranebounded packets. The fully keratinized cells, however, are packed with filaments 30 Å in diameter separated by an electron dense amorphous matrix, very similar to β-type keratin found in the feather rachis. The cells of the α layer, which is immediately below the β layer, contain 80 Å filaments very similar to the α-type keratin found in hair cortex and keratinizing stratified epithelia of mammals. Large quantities of glycogen are found in the cells of each layer during their genesis. Even though a stratum granulosum is not found underneath the α layer, the cells of the clear layer develop bodies which have histochemical and ultrastructural characteristics of keratohyalin granules. The old epidermis is then shed in toto at the junction of the clear layer (above) and the Oberhäutchen (below).
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    Cell & tissue research 206 (1980), S. 55-63 
    ISSN: 1432-0878
    Keywords: Phagocytosis ; Spermatozoa ; Ovum ; Fertilization ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Spermatozoa with intact acrosomes, as well as those coming into contact with the ovum at a smaller angle, and morphologically abnormal spermatozoa reach the plasma membrane of the ovum via an extensively dissolved zone of the inner layer of the vitelline membrane. This zone is assumed to be formed by overlapping of two or more tunnels formed by spermatozoa that had previously come into contact with the ovum. When a spermatozoon comes into contact with the plasma membrane of the ovum, many cytoplasmic processes extend outwards and cover it. Thereafter, the plasma membranes of the processes fuse, thereby phagocytizing the spermatozoon. It is assumed that the phagocytized spermatozoa cannot undergo transformation into male pronuclei and that they degenerate soon after phagocytosis.
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    Cell & tissue research 206 (1980), S. 225-232 
    ISSN: 1432-0878
    Keywords: Compound eye ; Photoreceptor membrane ; Electron microscopy ; Calcium-induced changes ; Artefacts ; Diptera
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary When the compound eyes of the fly Lucilia are fixed for electron microscopy with glutaraldehyde in common buffer solutions, artefactual whorls are liable to be formed from the photoreceptor microvilli. The whorls result from two factors: (i) a prolonged time interval prior to osmication, such as the “overnight” primary fixation or wash at 4° C commonly used in studies of compound eyes; (ii) as little as 1–2 mM Ca++ in the primary fixative and wash solutions. Osmication after short (1 h) glutaraldehyde fixation at 4° C, or omission of Ca++ and addition of 2 mM EGTA, prevent whorl-formation. In the tipulid fly Ptilogyna, similar artefacts are produced, but are confined to the distal zone of the microvilli that sheds during turnover.
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  • 94
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    Cell & tissue research 211 (1980), S. 83-93 
    ISSN: 1432-0878
    Keywords: Epithelium ; Cloaca ; Electron microscopy ; Hen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The epithela of the three divisions (coprodaeum, urodaeum, proctodaeum) of the cloaca of the hen, and of the excretory ducts (colon, ureter, vagina) which join the divisions, are described using light microscopy, and scanning and transmission electron microscopy. Each region of the cloaca has its typical epithelium. Special attention is focussed in this study on the boundaries between the different epithelia. The coprodaeal epithelium does not differ considerably from that of the colon; a transitional zone is not visible. Distinct border zones, however, are observed between the other regions (ureter — urodaeum; vagina — urodaeum and proctodaeum; urodaeum-proctodaeum; proctodaeum — cutis). Although the vaginal opening is generally thought to lie in the urodaeum, our investigations show that at the vaginal opening into the cloaca the ciliated epithelium changes, on one border to a secretory epithelium characteristic of the urodaeum and on the other border to that characteristic of the proctodaeum. These observations are discussed in relation to functional aspects.
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  • 95
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    Cell & tissue research 211 (1980), S. 171-174 
    ISSN: 1432-0878
    Keywords: Pancreatic D cell ; Neural control ; Vagotomy ; Electron microscopy ; Fowl
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In an attempt to determine the neural control of pancreatic D cells, the pancreatic islets of the domestic fowl were examined electron microscopically from 1 to 28 days after abdominal vagotomy. Exocytotic release of many secretory granules from D cells occurred one day after vagotomy. Rough endoplasmic reticulum developed and formed an arrangement of concentric whorls in the cytoplasm of D cells after axotomy. The altered D cells were also characterized by the occurrence of many peculiar dense bodies in the apical cytoplasm at all time periods studied. These bodies varied in shape and size, containing several round vesicles. The D cells were extensively depleted of granules after the longer time periods following vagotomy. The present results provide new morphological evidence for the vagus-nerve control of D cells, which may regulate the activity of islet cells.
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  • 96
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    Cell & tissue research 211 (1980), S. 191-206 
    ISSN: 1432-0878
    Keywords: Hypothalamus ; Transplants ; Vasopressin ; Median eminence ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Blocks of anterior hypothalamus were transplanted from 19 day-old fetuses of Wistar/Lewis rats into the third ventricle of adult male Brattleboro rats. Physiological changes in graft recipients and in sham-operated animals were monitored daily. Twenty days after surgery, the graft recipients and shamoperated animals were killed and their brains examined by correlative scanning and transmission electron microscopy. Host animals that exhibited both decreased polydipsia and increased urine concentration were found to have viable grafts within the third ventricle. The observed physiological changes suggested that synthesis and release of vasopressin occurred in the transplanted neurons. Grafts were well vascularized by vessels arising from the host hypothalamus. Neurons, with perikarya ranging from 8 to 30 μm in diameter, glial cells, and neurites were located throughout the transplants. A neurohemal contact zone, similar to that normally seen in the median eminence, could not be demonstrated in the grafts. The absence of complete glial and ependymal barriers indicates a relatively close association between cells in the transplants and the cerebrospinal fluid. A large increase in supraependymal neurons and their processes, including an eruption of neurons through the floor of the third ventricle in one animal, was observed in graft recipients but not in shamoperated animals.
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  • 97
    ISSN: 1432-0878
    Keywords: Carotid body ; Calcium ; Granular vesicles ; Exocytosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An attempt was made to study the subcellular localization of calcium in carotid body glomus cells of adult rats using fixation with glutaraldehyde followed by treatment with a mixture of pyroantimonate and osmium tetroxide. Precipitates were seen as electron-dense particles (EDP) in the glomus cells, mostly within membrane-bound organelles, such as dense-cored vesicles, mitochondria, small clear vesicles, multivesicular bodies, and especially in lysosomes. However, EDP were also seen in the nuclei and in the free cytoplasm of the glomus cells and even outside them. Preincubation of carotid bodies in media containing calcium and either high potassium or calcium-ionophore A 23187 resulted in a marked increase in the general precipitation pattern, there being an increased amount of EDP both in the glomus cell nuclei and in the cytoplasm. Dense-cored vesicles more often showed precipitates than those in the controls. Some dense-cored vesicles contained multiple precipitates, typically located in the electron-lucent area between core and vesicle membrane. Extensive diffusion of ions probably occurred during fixation before precipitation, making the localization of calcium and other precipitating cations unreliable. However, it is possible that precipitates, which were regularly seen in the dense-cored vesicles, may reflect the content of bound calcium. The possible significance of calcium in glomus cell function is discussed, and the need for more adequate methods is emphasized.
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  • 98
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    Cell & tissue research 234 (1983), S. 469-496 
    ISSN: 1432-0878
    Keywords: Bone marrow (human) ; Neutrophil granulocyte ; Granules ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Bone marrow from hematologically healthy adults was exposed to a number of fixation procedures for investigation of the heterogeneity of the granule population in neutrophil granulocytes at the ultrastructural level. Four main cell stages were distinguished: early promyelocyte, late promyelocyte, myelocyte, and mature neutrophil granulocyte and described separately; metamyelocytes and band-form or stab cells are described together. The characteristic changes in the cytoplasm during myelopoiesis were analysed quantitatively. Special attention was given to the development of the granule population. Three types of granule arise in successive cell stages: granules which develop a sub-structure in the matrix (nucleated granules) are formed in early promyelocytes, granules with a homogeneous electron-dense matrix (azurophil granules) in late promyelocytes, and granules with a less electron-dense matrix (specific granules) in myelocytes. The three types of granule remain present during myelopoiesis. The best results in distinguishing the granule types were obtained by prefixation either in 0.1% glutaraldehyde or in 1.5% glutaraldehyde followed by washing in phosphate-buffered Ringer solution to which aminotriazole had been added. Granule counts revealed for the mature neutrophil a total number of granules of about 220 per ultrathin section. This population of granules is composed of about 12% nucleated, 11% azurophil, and 77% specific granules. When our previous findings are taken into account, the existence of three successively formed and morphologically distinguishable types of granule in heterophil (neutrophil) granulocytes has been demonstrated for three mammalian species: the guinea pig, the rat, and man. A separate term for the early promyelocyte stage is proposed: eomyelocyte.
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  • 99
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    Cell & tissue research 234 (1983), S. 579-593 
    ISSN: 1432-0878
    Keywords: Rods ; Cones ; Retina ; Bullfrog ; Synapse ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Lumps of electron-dense material were observed in synaptic clefts associated with all types of photoreceptors, in the vicinity of the synaptic ribbons, in the retinae of dark-adapted frogs. Frogs were reared under a cyclic illumination (light on at 8:00; light off at 20:00) and then exposed to one of two courses of dark adaptation: one started from 11:00 in the morning, and the other started from 20:00 in the evening. The synaptic clefts of red rods became wider at some places where spherical or polygonal lumps of dense material were accumulated. The frequency and sectional area of the lumps increased faster for the first hour in the regime starting from 20:00 than in the regime starting from 11:00, then they reached the similar saturation levels of about 0.6 (per ribbon) and 1.6 to 1.8×104 (nm2) in both the regimes. In greenrod synapses, plate-shaped lumps of dense material were present in synaptic clefts and interspaces between the processes of second-order neurons. In cone synapses at the end of about 1 h darkness, the frequency and area of the lumps reached maximum values of about 0.12 (per ribbon) and 9×103 (nm2) in the regime starting from 11:00 and, about 0.08 (per ribbon) and 4 × 103 (nm2) in the regime starting from 20:00. On exposure to light, the dense material abruptly disappeared from all types of photoreceptor synaptic clefts. Large dense-core vesicles, occasionally observed in light-adapted rod photoreceptor terminals, seem to participate in exocytosis of the dense material. The number of dense-core vesicles per synaptic ribbon in a terminal was about 0.55 at the end of 3 h light in the morning and about 1.28 at the end of 12 h light in the evening. The increased number of dense-core vesicles during the daytime may contribute to the faster accumulation of dense material in the synaptic clefts. Although the chemical identification or the functional significance of the electron-dense material remains unknown, it is interesting that this material showed a rise and fall in response to darkness and illumination. Also the fact that this material is clearly visible will be helpful for future analysis of frog photoreceptor synapses.
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  • 100
    ISSN: 1432-0878
    Keywords: Ovulation (rabbit) ; Graafian follicle ; Perfusion ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Isolated ovaries from untreated, sexually mature rabbits were introduced into an in vitro perfusion system and perfused with a chemically defined medium containing albumin. The ovaries were perfused for up to 15 h (mean 11.5 h) and then processed for morphological investigation. Both at the light- and electron-microscopical levels, most of the ovaries exhibited a normal structure comparable with ovaries in situ. In two cases, however, marked accumulations of bacteria were found, although not inside the follicles. Since ovulation in the rabbit normally occurs between 9.5–13 h after mating or human chorionic gonadotrophin treatment, this model seems adequate for studies of ovulation in vitro. It is, however, important to study the ovaries microscopically after the perfusion to detect artifacts, e.g., bacterial infection, that may have influence on the process of ovulation.
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