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  • 1
    ISSN: 1432-0975
    Keywords: Key words Coral ; Calcification ; Biomineralisation ; Autoradiography ; Freeze-substitution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences
    Notes: Abstract  The uptake of 45Ca and/or 14C by the skeleton of coral colonies has been commonly used to investigate the processes of calcification. This study reports the differential uptake of these tracers within different regions of the skeleton and tissues of individual corallites and polyps of the hermatypic coral Galaxea fascicularis. Incubation in 45Ca in the light resulted in 80 percent of the 45Ca taken up being deposited in the skeleton. Autoradiography of transverse and longitudinal slices of freeze-substituted polyps and corallites showed that in the light 45Ca was incorporated into the exsert septa, the outside of the thecal walls of the corallite and the inner edges of the septa. Incorporation did not occur in the costae. The radioactivity in the skeleton was considerably greater than in the tissues. In the dark, or in the presence of the photosynthetic inhibitor Diuron, 45Ca was taken up by the exsert septa and was patchily distributed in the corallite walls which suggests that it was not a result of isotopic exchange. The differential incorporation of 45Ca onto the exsert septa was confirmed by scintillation counting. Negligible radioactivity remained in the extrathecal coelenteron after a brief 5 min rinse in non-radioactive seawater. Only 0.1% of 14C taken up in the light was incorporated into the skeleton and this was confirmed by autoradiography. In the presence of Diuron or in the dark, very little 14C was incorporated into tissues or skeleton and in autoradiographs was either not evident in the skeleton or the distribution was similar to that seen in autoradiographs of 45Ca uptake. These results show that the deposition of 45Ca, and therefore calcium carbonate, occurs at specific loci on the skeleton of a corallite. In the dark, deposition occurs specifically at the growing points of the corallite. Differential deposition of calcium carbonate within individual corallites has not been previously reported.
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  • 2
    ISSN: 1435-232X
    Keywords: Key words Moebius syndrome ; Calcification ; Arthrogryposis multiplex congenita ; Blood supply disturbance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We report the case of a Japanese girl with a severe type of Moebius syndrome. Her morphological features were a mask-like face, limitation of horizontal eye movements, severe bulbar palsy, multiple and bilateral arthrogryposis including the elbow, knee, and ankle joints, and clubfeet. After birth, her general condition became worse because of repeated apneic spells and aspiration pneumonias due to dysphagia. She finally required tracheotomy. Computed tomography (CT) of the brain revealed minute calcifications on the fourth ventricle floor; this may have been due to severe damage to the brain stem. It is most likely that the various manifestations in our patient were due to disturbance of the blood supply to arteries perfusing the brain stem and to some other arteries, at a critical stage of fetal development.
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  • 3
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    Aquaculture international 4 (1996), S. 237-252 
    ISSN: 1573-143X
    Keywords: Calcification ; Environmental disturbances ; Eumelanin ; Great scallop (Pecten maximus) ; Shell colour ; microstructure and melanization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Modifications of great scallop (Pecten moximus) shells have been noticed in many sites of scallop fisheries in Brittany, especially in shallow waters. These calcification abnormalities are linked to the appearance of a brown colouration of the internal calcified shell layer, due to the presence of a eumelanin associated with the insoluble organic matrix of the biocrystals. The appearance of this pigment generates many disturbances of the calcified foliated microstructure of the scallop internal shell layer. The mantle structure is not modified in brown shells as compared with white ones. No pathogenic signs such as hyperplasia or haemocytic infiltration have been observed. According to this observation, we hypothesize that the brown colour phenomenon is more a result of environmental disturbance rather than a symptom of a pathogenic disease. The colour abnormalities of the internal shell layer can be detected by a spectral analysis of its reflectance before it can be detected with the naked eye. This method, correlated to microstructural observations, gives a rapid and precise analysis of the appearance of the pigmentation on adult or juvenile scallops. It may be a useful method for the evaluation of the influence of environmental parameters, for example, on calcification and its abnormalities.
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  • 4
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    Calcified tissue international 56 (1995), S. 140-144 
    ISSN: 1432-0827
    Keywords: Bone sialoprotein ; Collagen ; Hole zone ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Bone sialoprotein (BSP) has an affinity to collagen fibrils [25]. A role of carbohydrate chains in the affinity was examined by removing sialic acids of BSP. Neuraminidase treatment of the BSP increased the binding to collagen. Binding sites of BSP on collagen were examined by biochemical and electron-microscopic methods. Purified bovine BSP was labeled with biotin. Collagen α chains or CNBr peptides were separated by electrophoresis and transfered to nitrocellulose membranes. The membranes were incubated with the biotin-labeled BSP, and the bound BSP was visualized with avidin conjugated with alkaline phosphatase. The labeled BSP was preferentially bound to the α 2 chain, and peptides derived from α 2 chain. In another experiment, the labeled BSP was incubated with reconstituted native collagen fibrils. The mixture was put on a copper grid, reacted with avidin conjugated with gold particles, and observed with an electron microscope. The gold particles were seen mainly within hole zones of the fibrils. BSP bound to the α 2 chain within the hole zones may regulate the onset of calcification at hole zones and the cell binding to collagen fibrils.
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  • 5
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    Calcified tissue international 56 (1995), S. 403-407 
    ISSN: 1432-0827
    Keywords: Bone sialoprotein ; Fibronectin ; Type I colfagen ; MC3T3-E1 cells ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Bone sialoprotein (BSP) containing an Arg-Gly-Asp cell-binding sequence was purified from bovine bone 4 M guanidine-HCl extract after HCl demineralization by a series of chromatographic procedures. When this protein was coated on culture dishes in the presence of type I collagen, it increased both DNA content and alkaline phosphatase (ALP) activity in osteoblast-like MC3T3-E1 cells, and stimulated calcification in the cells, whereas fibronectin, another cell-binding protein, showed a marked increase in the DNA content but had little effect on the ALP activity. These findings suggest that BSP is mitogenic for preosteoblasts and differentiating the cells into osteoblasts, thereby stimulating bone calcification
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  • 6
    ISSN: 1432-0827
    Keywords: Calcification ; Calcium phosphates ; Liposomes ; Mineralization ; Proteoglycans
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract A liposome-centered endogenous precipitation method was used to investigate the effect of ultrafilterable fragments from the enzymatic digestion of rat chondrosarcoma aggrecan on the formation of insoluble calcium phosphate salts in buffered solutions at pH 7.4 and 22°C. Unlike the intact aggrecan and its major chondroitin sulfate and core protein components, disaccharide units from chondroitinase degradation of the aggrecan and small (〈3kg/mol molecular weight) fragments from protease digestion of the core structure were found to be only weakly inhibitory toward mineral formation. Corresponding reductions in Ca2+-binding indicate that these fragments were unable to adsorb to active sites on the apatite surface for long enough periods to significantly hinder crystal growth. The data suggest that controlled enzymatic breakdown of aggrecan may be one possible mechanism by which the calcification of growth plate cartilage is allowed to advance in vivo.
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  • 7
    ISSN: 1432-0827
    Keywords: Osteoblasts ; Matrix proteins ; Collagen ; Cell differentiation ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Ipriflavone (IP), an isoflavone derivative, has been shown to interfere with bone remodeling by inhibiting bone resorption and perhaps stimulating bone formation. In this study, we have analyzed the effect of IP and its metabolites on the differentiation and function of human osteoblastic cells. Bone marrow stromal osteoprogenitor cells (BMC) and trabecular bone osteoblasts (HOB) were isolated from human donors. The former can be induced to differentiate by treatment with dexamethasone, whereas the latter represent a more differentiated osteoblast. Incubation of BMC with metabolite III (10-5 M) for 1 week induced modest but significant changes of alkaline phosphatase activity. Though both IP and metabolite III stimulated the expression of bone sialoprotein mRNA, a protein involved in cell attachment to the matrix, only metabolite III increased the steady-state level of decorin mRNA, a collagen fibrillogenesis-regulating proteoglycan. Metabolites III and V, but not the other isoflavones, increased the expression of type I collagen mRNA in HOB, whereas no detectable changes were observed in BMC cells with any of the experimental compounds. In HOB, an increased abundance of osteopontin and bone sialoprotein mRNA were also obtained after 1-week treatment with IP or metabolite V. No appreciable effects of IP or its metabolites were seen on osteocalcin expression and synthesis by either cell type. Finally, IP consistently increased the amount of 45Ca incorporated into the cell layer by BMC, and stimulated mineralization of both BMC and HOB, assessed by von Kossa staining. Thus, IP and its metabolites regulate the differentiation and biosynthetic properties of human bone-forming cells by enhancing the expression of some important matrix proteins and facilitating the mineralization process.
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  • 8
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    Protoplasma 181 (1994), S. 78-105 
    ISSN: 1615-6102
    Keywords: Calcification ; Coccolithophorids ; Coccolithogenesis ; Foraminifera ; Protists ; Taxonomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The diversity of calcified structures found in protists, the mechanisms utilized to form these structures, and the role these structures play in the taxonomy and systematics of the protists are presented. The two most frequently studied orders of protists which produce calcified structures, the coccolithophorids and foraminifera, are featured. However, consideration is given to the less known and least studied organisms.
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  • 9
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    Cell & tissue research 275 (1994), S. 345-353 
    ISSN: 1432-0878
    Keywords: Osteogenesis ; Ossification ; Mineralization ; Calcification ; Cell necroses ; Mouse (NMRI)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Previous investigations concerned with in vitro osteogenesis and mineralization have revealed some indication of a participation of cell necroses in the course of calcification. These observations were confirmed by in vivo investigations on desmoid ossification in fetal mouse calvariae, where abundant necrotic osteoblasts were found at the mineralization border and in the osteoid. In the present study, ossification of long bone cortices from fetal mice was investigated by use of electron microscopy. Specimens obtained from the collection of the Institute of Anatomy, Free University of Berlin (mouse fetuses, forearm; rat fetuses, forearm) were reinvestigated for control purposes. In all cases, mineralization of osteoid was accompanied by cell necroses. Cell degeneration was characterized by swelling of the endoplasmic reticulum and loss of the plasma membrane resulting in freely distributed vesicular structures. Cell debris was incorporated within the mineral. Initially, cell necroses in the perichondrium occurred in the region surrounding the hypertrophic cartilage and the matrix of which showed spots of endochondral mineralization. Necrotic osteoblasts occurred simultaneously with mineralization of the osteoid. During further ossification of the long bone cortices, the number of necrotic cells increased markedly. In addition to necrotic cells, healthy osteoblasts, osteocytes and perichondral tissue were present, indicating that an artifact can be excluded. The importance of cell necroses in the process of mineralization is as yet unclear. Possibly, the cells act as calcium and/or phosphate stores, which are liberated by cell death to increase the amount of mineral constituents at sites of mineralization.
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  • 10
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    Calcified tissue international 52 (1993), S. 94-98 
    ISSN: 1432-0827
    Keywords: Atherosclerosis ; Calcification ; Hydroxyapatite ; Cholesterol ; Filipin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Cholesterol and calcium phosphate, the latter in the form of hydroxyapatite, accumulate in atherosclerotic lesions. In this report, we demonstrate that these organic and inorganic constitutents of lesions can accumulate together, closely associated in crystal agglomerates. Using the fluorescent cholesterol probe, filipin, we identified unesterified cholesterol that was associated with calcium granules in tissue sections of lesions. We also have shown that small crystallites of cholesterol can associate with preformed hydroxyapatite crystals in vitro. Scanning electron microscopy couple with energy-dispersive X-ray analysis demonstrated the physical association of many small crystallites of cholesterol with larger crystals of hydroxyapatite. These small crystallites of cholesterol associated with hydroxyapatite stained with filipin. This contrasted with the lack of filipin staining of unassociated larger cholesterol crystals or hydroxyapatite alone. How cholesterol and calcium come to be closely associated in crystal agglomerates within atherosclerotic lesions remains to be determined.
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  • 11
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    Cell & tissue research 272 (1993), S. 321-327 
    ISSN: 1432-0878
    Keywords: Calcification ; Calcinosis ; Calculi ; Microliths ; Salivary gland calculi ; Salivary glands ; Cat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract This investigation concerns the natural history of microlith in the salivary glands of cat. Microliths were detected in more sublingual than submandibular glands and were almost absent in the parotid. They were found intraparenchymally, intraluminally and interstitially, and ultrastructurally in phagosomes of acinar, ductal and myoepithelial cells, intermixed with the cytoplasm of degenerate acinar cells, and in intraparenchymal macrophages and a multinuclear giant cell. They appear to form in healthy acinar cells during autophagocytosis, and possibly to be discharged luminally, laterally or basally, and to form in the debris of degenerate cells intraparenchymally and intraluminally. They appear to be removed by expulsion in the saliva, scavenging macrophages, and possible eventual degradation in the parenchymal phagosomes. The greater occurrence of microliths in the sublingual gland may relate to a low level of secretory activity, and the near absence of microliths in the parotid to a low level of calcium. The feline salivary glands were found to be an outstanding model for the investigation of microlithiasis.
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  • 12
    ISSN: 1432-0878
    Keywords: Spicule ; Calcification ; Organic matrix ; Collagen ; Leptogorgia virgulata (Cnidaria)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Spicules of the gorgonian Leptogorgia virgulata possess an insoluble matrix fraction that is predominantly collagenous in summer months. This collagenous component is largely absent in winter months. Using an antibody directed against the 140 kD collagenous protein (CP) of the insoluble matrix, immuno-gold labelling was employed to localized this protein at the transmission electron-microscopy level throughout the year, and in different areas of the gorgonian colonies. Within the tip regions, the 140 kD CP varied throughout the year in the spicules, electron-dense bodies (EDBs) of scleroblasts, polyp vesicles, desmocytes and axes. In the mid and base regions, the 140 kD CP varied throughout the year in the spicules, EDBs and lysosomes of scleroblasts, desmocytes and axes. This variation in the location and density of the label suggests a dynamic annual cycling of the collagenous component of the insoluble matrix. EDBs may transport a collagenous component of the matrix to the spicule-forming vacuole. A component of the 140 kD CP may be transported and/or degraded by polyp vesicles and lysosomes, respectively. The pattern of labelling of the axial region suggests that translocation and storage of a component of the collagenous protein may occur. Environmental factors may be responsible for the triggering of matrix cycling.
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  • 13
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    Cell & tissue research 272 (1993), S. 533-543 
    ISSN: 1432-0878
    Keywords: Mucocytes ; Calcification ; Intercellular spaces ; Zooxanthellae ; Calicoblastic ectoderm ; Confocal laser scanning microscopy ; Freeze-substitution ; Corals: Galaxea fascicularis, Acropora formosa, Tubastrea faulkneri (Cnidaria)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The extra-thecal epithelia of cryofixed undecalcified, freeze-substituted polyps of the scleractinian corals Galaxea fascicularis and Tubastrea faulkneri and axial and basal polyps of Acropora formosa have been examined, in anhydrously prepared thick slices, by confocal laser scanning light microscopy. The avoidance of chemical fixation and decalcification makes it possible to determine whether previously seen structures are real or artefactual products of swelling, shrinkage and distortion. All of the epithelia of all the corals examined are characterised by well defined intercellular spaces. Mucocytes are present in all cell layers in Galaxea and Tubastrea but are not present in any cell layers in the axial polyp of Acropora although they are abundant in the oral ectoderm of the basal polyps in this coral. Zooxanthellae are absent in Tubastrea, the epithelia of the exert septa of Galaxea and the axial polyp of Acropora. The calicoblastic ectoderm is generally composed of thin squamous cells with large intercellular spaces. At rapidly calcifying regions such as the tips of the exert septa of Galaxea, the calicoblastic cells are elongated with extensive arborisation of the basal regions of the cells. They are separated by large intercellular spaces and contain numerous fluorescent granules. The apical regions of these cells appear to be closely applied to the surface of the skeleton. There is no evidence of a space between the apical region of the calicoblastic cells and the skeleton.
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  • 14
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    Calcified tissue international 50 (1992), S. 134-136 
    ISSN: 1432-0827
    Keywords: Calcification ; Dentinogenesis ; Odontoblast ; Calcium transport ; Ion-selective microelectrode ; pH
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Ca2+ ion activities and pH were measured in the odontoblast/predentin region of rat incisors by means of the microelectrode technique. In Ringer solution, the apparent resting membrane potential of odontoblasts was determined to be -24±4 mV (mean±SE), whereas the odontoblast intracellular pH was found to be 6.66±0.02. The values obtained are within the range of other cell types, as measured in similar incubating solutions. The pH in the extracellular predentin was higher than the intracellular pH, 7.00±0.02. The Ca2+ ion activity in predentin (pCa=2.94±0.15) was found to be significantly (P〈0.001) higher than that in the dental pulp extracellular fluid (pCa=3.37±0.14). The 2–3 times higher calcium activity extracellularly in predentin, compared with the dental pulp, implies the existence of some ion-concentrating mechanism across the odontoblast layer in the direction of the mineralization front.
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  • 15
    ISSN: 1432-0827
    Keywords: Liposomes ; Hydroxyapatite ; Aggregation ; Atherosclerosis ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Some stages in the calcification of atherosclerotic plaques may involve associations between lipids and hydroxyapatite (HA) by surface interactions. Liposomes, artifical membranous lipid vesicles, have been used in this study as model structures for biological calcification processes. Liposome (containing cholesterol and phosphatidylcholine in most cases) suspensions were prepared by sonication, after which HA seed crystals were added to the suspensions and stirred at 37°C. Aliquots of the liposome suspensions were analyzed for particle size distribution and by transmission electron microscopy and electron diffraction. The results showed that HA induced aggregation of liposomes and modifications of the microscopic shapes of the liposomes in the aggregates. These data can be explained by the electron diffraction pattern where superimposition of liposome reflection and crystal reflection exists and may suggest organic-inorganic interaction. The potential of HA crystals to induce formation of liposome aggregates may be seen as a step in atherosclerotic plaques calcification.
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  • 16
    ISSN: 1432-0827
    Keywords: Growth plate ; Collage ; Proteoglycans ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Calcification occurs in the extracellular matrix of the hypertrophic zone of the growth plate when the extra-cellular matrix volume is reduced to a minimum and alkaline phosphatase content is maximal. The present study shows that significant quantitative and qualitative changes occur in the composition and structure of macromolecules in the extracellular matrix before and during calcification in the proximal tibial growth plate of the bovine fetus. These were detected in part by using microchemical and microimmuno-chemical analyses of sequential transverse frozen sections at chemical analyses of sequential transverse frozen sections at defined sites throughout the growth plate. Concentrations of matrix molecules in the extracellular matrix have not previously been determined biochemically. They were measured per unit matrix volume by using combined immunochemical/chemical-histomorphometric analyses. The concentrations within the extracellular matrix of the C-propeptide of type II collagen, aggregating proteoglycan (aggrecan), and hyaluronic acid all progressively increased in the maturing and hypertrophic zones, being maximal (or near maximal) at the time of initiation of mineralization. These results for proteoglycan are contrary to some earlier reports of a loss of proteoglycan prior to mineralization which measured the tissue content of proteoglycan rather than that present in the extracellular matrix, the volume of which is progressively reduced as the growth plate matures. The C-propeptide data provides a quantitative confirmation of previous immunohistochemical studies. Total collagen concentration (measured as hydroxyproline) in the extracellular matrix initially increased through the proliferating and maturing zones but then rapidly decreased in the hypertrophic zone. Immunohistochemical studies revealed that this is associated with the unwinding of the triple helix of type II collagen (previously shown to result from cleavage) which starts in pericellular sites in the zone of maturation (when type X collagen is first synthesized) and then extends throughout the hypertrophic zone. The significance of these matrix changes in the development and mineralization of the growth plate is discussed.
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  • 17
    ISSN: 1432-0827
    Keywords: Osteoblasts ; Collagen ; Cross-links ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The early stage of cross-link formation in bone collagen was studied in a cell culture system. An osteogenic cell line that produces and accumulates a remarkably high amount of collagen, and that eventually forms bone-like structures, was used in this study for its time-dependent development of reducible cross-links. It was found that precursors of the cross-link, dehydro-dihydroxynorleucine and dehydro-hydroxynorleucine became detectable as soon as the cells attained a confluent state. They showed maximal amounts at day 3–5 after confluence, but substantially disappeared at day 10 after confluence. In contrast, two characteristic cross-links of bone collagen, dehydrodihydroxylysinonorleucine dehydro-DHLNL and dehydrohydroxylysinonorleucine (dehydro-HLNL), which were present in trace amounts at the stage of cell confluence, gradually increased in amount and reached a plateau at day 10, just when their precursors disappeared. Thus, it was found that there was a time lag of about a week between the maximal formations of precursors and cross-links of bone collagen in this system. The significance of this time lag was interpreted in terms of the minimum essential accumulation of collagen for the precursor-product transition. The ratio of dehydro-DHLNL to dehydro-HLNL was as low as 0.7 at day 3 after confluency, increased to 4.2 at day 20, the period just before mineralization began, and decreased thereafter, suggesting a qualitative change in bone collagen associated with mineralization.
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  • 18
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    Calcified tissue international 50 (1992), S. 553-563 
    ISSN: 1432-0827
    Keywords: Mineralization ; Calcification ; Dystrophic calcification ; Bone organ culture ; Chick embryo
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The effectiveness of Na-β-, and Ca-glycerophosphates (GPs) in inducing mineralization was tested during long-term organ culture of femurs from 14-day-old chick embryos. When bones were incubated with Na-GP, a 66% rise in inorganic phosphate level was measured in the medium, supporting the notion that provision of a substrate for alkaline phosphatase (ALP) increased available phosphate. On the other hand, if the concentrations of Ca2+ were raised, available inorganic phosphate was decreased. Similarly, increases in inorganic phosphate decreased available calcium. Both GPs induced mineralization in bone and cartilage, but more matrix was mineralized with Ca-GP. However, the induction of mineralization by GPs was accompanied by dystrophic calcification, reduction of matrix formation and ALP activity, and increased release of lactate dehydrogenase into the culture medium. The new osteoid, which formed during culture, mineralized in the absence of GPs without the above adverse effects provided the culture period was longer than 15 days. The described organ culture system therefore facilitates studies of the mechanism of bone mineralization without the disadvantages of GP addition.
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  • 19
    ISSN: 1615-6102
    Keywords: Calcification ; Calcium carbonate ; Rhodogorgon ; Red algae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Calcification inRhodogorgon carriebowensis J. Norris et Bucher was associated with a particular cell type in the cortex. Calciferous cells were 4–6 times the length of cortical assimilatory cells. The distal two-thirds of the calcifying cell was invested with a thick wall that stained with periodic acid Schiff. Thick fibrils formed a reticulum and surrounded grains of calcium carbonate that ranged in shape from rhombohedral to subspherical and were up to 200 nm in greatest dimension. The proximal third of the cell was a tapering uncalcified stalk. The narrow base of the cell was attached to the subtending cell of the fascicle by a normal septum with a pit plug. The cell within the calcified wall matrix was usually flattened and had a very small volume. Cellular contents were dense; even when organelles could be discerned, they could not be identified. X-ray microanalysis revealed that other elements commonly found mixed with calcium carbonate are virtually absent from mineral deposits inR. carriebowensis, but electron diffraction study showed d-spacings that varied from those of pure calcite. Current models of red algal calcification are discussed in light of the findings on this alga.
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  • 20
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    Primates 32 (1991), S. 265-268 
    ISSN: 0032-8332
    Keywords: Incisor crown formation ; Calcification ; Enamel ; Japanese macaques ; Macaca fuscata ; Sexual variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Fifteen laboratory-born Japanese macaques,Macaca fuscata (Blyth, 1875), were examined radiographically for the timing of initial crown calcification of the permanent upper first incisors. The mean age of initial calcification was 199.8 days in females and 204.7 days in males; the sexual difference was significant (p〈.05). Precocious incisor calcification in females inM. fuscata resembles that inM. nemestrina andHomo sapiens.
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  • 21
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    Journal of comparative physiology 161 (1991), S. 413-418 
    ISSN: 1432-136X
    Keywords: Chitin ; Aragonite ; Calcification ; Crystalline shape ; Anodonta cygnea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary An orthorhombic structure α-chitin, probably in the form of a chitin-protein complex, was identified in the matrix of the shell of Anodonta cygnea by X-ray diffraction. Aragonite crystals of pseudohexagonal symmetry were also found by a Lauegram on the nacreous layer of the shell. The orthorhombic structure of these two compounds together with the identical reticular spacing d110 corroborate, in Anodonta cygnea, the indirect chitin-aragonite relationships already suggested for molluscan shells. Observations with SEM in the inner surface of the shell showed CaCO3 crystals with irregular geometrical shapes in spring and summer and regular geometrical shapes in autumn and winter. The more elaborate aspect appearing in winter corresponds to an accurate hexagonal shape. This suggests that the observed variability may depend on the balance between calcium and hydrogen ions in the extrapallial fluid.
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  • 22
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    Cell & tissue research 243 (1986), S. 91-99 
    ISSN: 1432-0878
    Keywords: Teeth ; Calcification ; Adenosine triphosphatase ; Calcium-alkaline phosphatase ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Enzymatic activities of calcium-magnesium dependent adenosine triphosphatase (Ca-ATPase) and nonspecific alkaline phosphatase (ALPase) were localized at the initial calcification sites of dentin and enamel of rat incisor teeth using electron-microscopic cytochemistry. Ca-ATPase was localized in the Golgi cisternae, cytoplasmic vesicles and along the outer surface of the presecretory and secretory ameloblasts, whereas it was totally absent from the odontoblasts in the pulp. Inversely, ALPase reaction was localized along the outer surface of the odontoblasts, but almost completely absent from the ameloblasts. Diffuse extracellular reactions of both enzymes were distributed throughout the unmineralized fibrous matrix of mantle dentin in which a large number of matrix vesicles were scattered. Both Ca-ATPase and ALPase reactions, which appeared in the matrix vesicles in the process of formation of mantle dentin, became most conspicuous at the site of initial dentin calcification. At this stage, an intense Ca-ATPase reaction also appeared along some of the collagen fibrils adjacent to the reactive matrix vesicles. No ALPase reaction was localized along these Ca-ATPase reactive collagen fibrils. Our observations suggest strongly that Ca-ATPase in the matrix vesicles originates from the inner enamel epithelium and/or preameloblasts whereas ALPase originates from the odontoblasts in the pulp. The importance of the coexistence of both enzymes for the control of initial calcification of dental hard tissues is suggested.
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  • 23
    ISSN: 1432-0878
    Keywords: Ca2+ transport ; Calcification ; Autoradiography ; Gorgonians ; Leptogorgia virgulata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The route of calcium transport to the sites of spicule formation in the gorgonian Leptogorgia virgulata has been examined by the use of 45Ca as a tracer in light- and electron-microscopic autoradiography. From 1 to 15 min after the 15-min incubation the tracer accumulates in the axis. After 15 min there is a movement of label out of the axis largely to the peripheral region of the axis, the axial epithelium, and the mesoglea. By 60 min much of the label is in the spicules reaching its maximum level at 120 min. When calcium enters the scleroblast from the mesoglea, it appears to be transported to the spicule by electron-dense bodies. There does not appear to be a simultaneous release of all ionic calcium from the axis, but rather a continuously increasing efflux which levels off at 60–120 min. Not all of the calcium reaching the axis will traverse it en route to spicules; instead, a portion of it apparently precipitates as an amorphous compound.
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  • 24
    ISSN: 1432-0827
    Keywords: Phosphoprotein ; Dentin ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Free and matrix-bound phosphophoryns, both highly phosphorylated proteins in dentin, were prepared from EDTA extract and CNBr-digests of bovine dentin. The two components were purified by DEAE-cellulose, SP-Sephadex, and gel filtration chromatography. The matrix-bound component was eluted as a distinct peak from the free component in the above chromatographic systems. Amino acid composition of the purified matrixbound component indicated that this component consisted of phosphophoryn and collagen in the ratio of 2:3 based on the number of the residues. The matrix-bound component could not be reconstituted by mixing phosphophoryn with collagen CNBr peptides. Artificial crosslink products of free phosphophoryn and collagen CNBr-peptides by the carbodiimide method showed similar properties to the physiological matrix-bound phosphophoryn. The bond between phosphophoryn and collagen of the matrix-bound component is assumed to be a covalent crosslink.
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  • 25
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    Calcified tissue international 36 (1984), S. 332-337 
    ISSN: 1432-0827
    Keywords: Proteolipid ; Phospholipids ; Vitamin D ; Calcification ; Cartilage
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Previous studies have shown thatin vitro calcification of chick epiphyseal cartilage matrix vesicles is proteolipid-dependent. The purpose of this research is to examine the role of proteolipid in cartilage calcificationin vivo by comparing the proteolipid concentration of normal and vitamin D-deficient chick epiphyseal cartilage, the relationship of proteolipid to other tissue lipids, and its ability to supportin vitro apatite formation. Proteolipid was isolated from the upper growth centers (reserve cell zone, upper proliferative zone) and lower growth centers (lower proliferative, hypertrophic, and calcified cartilage zones) of long-bone epiphyses from 3-week-old normal and rachitic male white leghorn chicks by Sephadex LH-20 chromatography of the crude phospholipid component of the total lipid extract. In both normal and rachitic tissue the proteolipid/dry weight and proteolipid/total lipid ratios were greater in the lower growth center than in the upper zones. No statistically significant change in the proteolipid/total lipid ratio in rachitic tissues relative to comparable cell zones in normal cartilages was observed. However, there was an increase in the nonproteolipid phospholipid content of rachitic tissues, altering the relative proteolipid/phospholipid composition. Whereas proteolipids from normal tissue supportedin vitro calcification, proteolipids from rachitic tissues did not, indicating a direct effect of vitamin D on proteolipid structure. These data support the hypothesis that failure of rachitic cartilage to calcifyin vivo may be due in part to alterations in phospholipid and proteolipid metabolism.
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  • 26
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    Calcified tissue international 36 (1984), S. 214-218 
    ISSN: 1432-0827
    Keywords: Proteolipid ; Calcium-phospholipid-phosphate complexes ; Calcification ; Hydroxyapatite ; Membranes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary This study demonstrates that calciumphospholipid-phosphate complexes (CPLX) and calcifiable proteolipid are associatedin vivo by establishing that they can be co-isolated from calcified bacteria. Both of these membrane constituents, which support apatite formationin vitro, have been isolated independently fromBacterionema matruchotii. However, isolation of proteolipid was preceded by demineralization in 2N formic acid, thereby dissociating bound Ca, whereas isolation of CPLX included sonication of calcified bacteria in 2:1:1.5 chloroform:methanol:Tris buffer, thereby dissociating any protein. Co-isolation is possible by demineralizing the calcified bacteria with 50 mM phthalic acid, pH 5.5, followed by extraction with 2:1 chloroform:methanol, and precipitation of crude phospholipid with acetone. CPLX and proteolipid are present in all Sephadex LH-20 chromatographic fractions of the crude phospholipid and of diethyl ether precipitates of the crude phospholipid. CPLXs contain protein:phospholipid:Ca:Pi but differ in relative composition from each other and from independently isolated CPLX. The Ca:phospholipid:Pi molar ratio of diethyl ether precipitable proteolipid-CPLX is most similar to previously published values for CPLX. The protein content of CPLX accounts for all of the proteolipid apoprotein in each Sephadex LH-20 fraction.
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  • 27
    ISSN: 1432-0878
    Keywords: Gorgonians ; Organic matrix ; Autoradiography ; Calcification ; Leptogorgia virgulata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The sequence of the synthesis and transport of the organic matrix of spicules has been elucidated in the gorgonian Leptogorgia virgulata by use of 3H-aspartic acid as the tracer in electron-microscopic autoradiography. The entire process of matrix synthesis and transport takes approximately 2 h. It seems that the protein moiety of the organic matrix is synthesized in the RER prior to 5 min following the initial 10 min incubation in the tracer. At the 5 min chase the label is moving from the RER to the Golgi complexes where the carbohydrate moiety of the matrix is presumed to be synthesized. At the 5 to 15 min chases the label is transported out of the Golgi complexes via Golgi vesicles. This phase continues for 30 min. From 60 to 120 min the 3H-aspartic acid moves to the spicules. After 120 min the majority of the label has moved into the spicules. Silver grain counts over both multivesicular and electron-dense bodies remain at relatively low and constant levels over 4 h indicating that neither organelle is involved in the synthesis and transport of the organic matrix.
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  • 28
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    Cell & tissue research 236 (1984), S. 413-420 
    ISSN: 1432-0878
    Keywords: Calcification ; Crab cuticle ; Cell coat glycoprotein ; Carbonic anhydrase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In the crab cuticle the interprismatic septa (IS), which correspond to imprints left in the cuticle by the margins of the epidermal cells, penetrate the twisted structure of the chitin-protein matrix. The ultrastructure and geometric relationship between the fibrous architecture and the pattern of the prisms is described. The cytochemical characterization of the IS, by pronase treatment and ruthenium red staining, supports the hypothesis that this material corresponds to cell-coat glycoproteins released in the cuticle during secretion of the organic matrix. Calcification begins after ecdysis in the external laminae of the pigmented layer and along the IS. The presence of cation-binding glycoproteins in the sites where calcification is initiated could induce the nucleation of the mineral phase by concentrating calcium. The extracellular distribution of carbonic anhydrase, which favours carbonate deposition, is observed on ultrathin sections over the IS.
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  • 29
    ISSN: 1432-0827
    Keywords: Matrix vesicles ; 32P-phosphate and45Ca-metabolism ; Epiphyseal cartilage ; Calcification ; Alkaline phosphatase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Matrix vesicle-enriched fractions were isolated from different zones of epiphyseal cartilage by nonenzymatic methods involving tissue homogenization, differential centrifugation, and isosmotic Percoll gradient fractionation. Uptakes of both32Pi and45Ca were studied concomitantly over periods from 20 min to 24 h. Percoll density gradients separated epiphyseal microsomes into two alkaline phosphatase-rich fractions: a low-density noncalcifiable fraction (P-I), and a higher-density fraction (P-II) which readily mineralized. The P-II fraction was found only in calcifying regions of the growth plate. Based on chemical and physical properties and enzyme activities, both fractions were similar except that P-II contained significantly higher levels of mineral ions than did P-I, and had lower levels of alkaline phosphatase. The mineral appeared to be primarily in a noncrystalline form. Metabolism of32Pi and45Ca by P-II followed a complex kinetic pattern in which accumulation of large amounts of both ions was preceded by an initial limited burst of uptake and a lag-phase of variable duration. During mineral ion loading, the density of the P-II fraction progressively increased as evidenced by co-migration of45Ca,32Pi, and alkaline phosphatase to increasingly higher densities. During the period of early mineral deposition (1–5 h), Ca/P uptake ratios were very low (1.0–1.2) and X-ray diffraction patterns showed a predominantly amorphous pattern. This suggests that the mineral accumulated in matrix vesicles is initially some form of noncrystalline calcium monohydrogenphosphate. L-tetramisole, a potent inhibitor of alkaline phosphatase, inhibited accumulation of both45Ca and32Piin the absence of organic P substrates,32Pi being preferentially inhibited over45Ca. This finding, coupled with recent studies on the behavior of alkaline phosphatase at physiological pH, suggests that the protein is not acting as a phosphohydrolase, but rather as a Pi-binding or transport agent in vesicle-mediated calcification.
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  • 30
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    Calcified tissue international 35 (1983), S. 723-727 
    ISSN: 1432-0827
    Keywords: Calcification ; Scleroderma ; CRST syndrome ; High resolution TEM ; Microanalysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary This paper reports the study of a subcutaneous heterotopic calcification from a patient with Thibierge-Weissenbach's syndrome, a type of creeping scleroderma included in the CRST syndrome. These local deposits, whose origin is still unknown, are commonly considered to be a classic apatite phase. Using SEM, high resolution TEM, electron diffraction, infrared spectrometry, and SEM and TEM microanalysis, it is demonstrated that this material is highly heterogeneous and appears in a nonstoichiometric, carbonated, calcium ion-deficient apatitic solid phase. Our study shows the co-existence of dense globules presenting an ill-organized, more or less amorphous phase (ACP) or microcrystalline (OCP, β tricalcium phosphate), with scattered apatite crystals, and of interglobular apatite crystals with a good cristallinity.
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  • 31
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    Calcified tissue international 35 (1983), S. 783-790 
    ISSN: 1432-0827
    Keywords: Amorphous calcium phosphate ; Apatite ; Calcification ; Hydrolysis ; Octacalcium phosphate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The hydrolysis of previously prepared amorphous calcium phosphate (ACP) was studied in a solution “saturated” with ACP; this eliminated the initial consumption of acid due to ACP dissolution. The procedure established that conversion of a high-concentration ACP slurry to an apatite involves two processes: the first process consumes acid and indicates the formation of a more acidic calcium phosphate intermediary with the solubility of octacalcium phosphate (OCP); the second process consumes base and indicates the conversion of the intermediary to apatite and, possibly, direct conversion of ACP to apatite. The thermodynamic analysis of the solution composition data suggests that ACP converts into a nonstoichiometric apatite when the OCP-like intermediary is formed, and a stoichiometric apatite is formed when no OCP-like intermediary is involved.
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  • 32
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    Calcified tissue international 35 (1983), S. 48-55 
    ISSN: 1432-0827
    Keywords: Enamel ; Density centrifugation ; Hydroxyapatite ; Carbonate ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Whole enamel scrapings from unerupted teeth of embryonic calves have been separated into fractions of varying density by stepwise centrifugation in bromoform-toluene mixtures of increasing specific gravity. Partition of enamel in this manner yields individual fractions of increasing mineral phase age and maturation. Whole scrapings and isolated fractions of the fetal bovine enamel were examined by X-ray powder diffraction, scanning electron microscopy, and atomic absorption and infrared spectroscopy to determine time-related changes in the physicochemical nature of the constituent mineral phase particles. These analyses showed poorly crystalline hydroxyapatite (HA) as the only detectable solid phase of calcium phosphate present in all fractions, its degree of crystallinity increasing with increasing density. Molar Ca/P ratios and magnesium content were highest in lowest density fractions. Carbonate vibration bands at 875 and 1420–1450 cm−1, common to mineralized tissue, were observed in intermediate and higher density fractions and in whole unfractionated enamel. Another carbonate band at ∼705 cm−1, unusual to vertebrate calcified tissue, was detected in low density fractions and disappeared rapidly with increasing enamel maturation. Its precise relation with the enamel mineral phase has not been determined.
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  • 33
    ISSN: 1432-0827
    Keywords: Calcification ; Glycosaminoglycan ; Glycoprotein ; Collagen ; Acid phosphatase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Glycosaminoglycans (GAGs) and glycoproteins are essential components for osteogenesis. We have examined rat osteoblasts, osteoid, transitional zone, and fully calcified bone matrix, utilizing Spicer's high-iron diaminethiocarbohydrazide-silver protein (HID-TCH-SP) method for sulfated glycoconjugates and Thiéry's periodate-TCH-SP (PA-TCH-SP) method for vicinal glycol-containing glycoconjugates. HID-TCH-SP stained cytoplasmic granules of osteoblasts. Stain deposits in the extracellular matrix were observed in decreasing amounts in osteoid, the transitional zone, and fully calcified bone matrix. Enzyme digestion with testicular hyaluronidase removed most HID-TCH-SP stain deposits. PA-TCH-SP staining was observed with increasing intensity in rough endoplasmic reticulum, Golgi saccules, and cytoplasmic granules. Collagen fibrils in osteoid were weakly stained with PA-TCH-SP, and their staining appeared even weaker in fully calcified bone matrix. In contrast, collagen fibrils in calcified cartilage stained intensely with the PA-TCH-SP method. Focal circular profiles (0.1–0.5µm in diameter), which lacked collagen fibrils but reacted moderately with PA-TCH-SP, were frequently seen in the transitional zone and fully calcified bone matrix, but were only occasionally present in osteoid. The presence of testicular hyaluronidase-resistant GAG and acid phosphatase in these focal areas suggests that they represent sites of GAG degradation. The eventual loss of HID-TCH-SP staining in the bone matrix suggests that removal of sulfated glycoconjugates may be a requisite for expansion of initial calcification sites and/or complete calcification.
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  • 34
    ISSN: 1615-6102
    Keywords: Calcification ; Coccolithophorids ; Polysaccharide localization ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Emiliania huxleyi is a marine coccolithophorid which produces coccoliths,i.e., particles consisting of calcite and macromolecular organic material. The coccoliths are formed intracellulary in specialized organelles which comprise a coccolith vesicle (CV) and a reticular body (RB), together forming the CV/RB system or calcifying system. After termination of calcification, the coccolith is extruded and incorporated into the coccosphere,i.e., one or several layers of extracellular coccoliths surrounding the cell. Apart from the coccolith-producing cells (C cells) ofE. huxleyi, there are naked cells (N cells) which seem to have lost the capacity to produce coccoliths but are very similar to the C cells in other morphological respects. Biochemical studies have revealed that polysaccharides may play a regulatory role in calcification. The aim of the present study was to determine the localization of polysaccharides in both C and N cells electron microscopically. For this purpose, a cytochemical staining technique according toThiéry (1967) was applied. The CV/RB system of C cells was conspicuously stained. Due to the excellent stainability of this system, a putative succession of morphological stages during coccolithogenesis could be described. The staining pattern of the N cells closely resembled that of the C cells. It was found, however, that the “calcifying” system of N and C cells differed in both morphology and position. It is suggested that the divergent morphology of the “calcifying” system of N cells accounts for its failure to produce coccoliths.
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  • 35
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    Calcified tissue international 34 (1982), S. 428-430 
    ISSN: 1432-0827
    Keywords: Epiphyseal cartilage ; Carbonic anhydrase ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The distribution of carbonic anhydrase isoenzymes C and B in the rat epiphyseal growth cartilage was demonstrated by an immunohistochemical method. The isoenzymes were found in different locations. Isoenzyme C was in the extracellular matrix of the hypertrophic and calcifying cartilage, and no reaction was observed in the chondrocytes. In contrast, the antiserum against isoenzyme B revealed only a weak cellular staining. This supports the hypothesis that carbonic anhydrase isoenzyme C, which is the high-activity form, changes the pH in the extracellular fluid of calcifying cartilage, favoring the deposition of calcium phosphate.
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  • 36
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    Calcified tissue international 34 (1982), S. 382-390 
    ISSN: 1432-0827
    Keywords: Avian osteopetrosis ; Avian oncornavirus ; Ultrastructure ; Calcification ; Bone cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Diaphyseal tibial bone of 12.5 – 13-day and 19-day-old embryos and 20-day-old hatched chicks infected with retrovirus MAV.2-O were examined by transmission electron microscopy. The viruses were associated with lining osteoblasts and osteocytes. Whereas the infection of the osteoblast layer seemed to be a transient stage, virus association with osteocytes was a constant and main ultrastructural feature. The viruses were found either in the osteoid or in the periosteocytic space of the bone lacunae. They arose from dense cytoplasmic areas located near the cell plasmalemma via a budding process. The newly budded virus particles often had a large tail or a fine stalk-like process lost in the extracellular space. The viruses underwent calcification by deposition of inorganic material and were incorporated in the bone trabeculae. No production of virus was observed in typical osteoclasts with well-differentiated ruffled borders. The viral-induced avian osteopetrosis seemed to result from increased bone deposition through stimulation of osteoblast and osteocyte activities, whereas osteoclastic bone resorption seemed to be undisturbed.
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  • 37
    ISSN: 1432-0878
    Keywords: Gorgonians ; Spicule ; Scleroblast ; Calcification ; Leptogorgia virgulata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Ultrastructural examination of original and regenerated branch tips of the gorgonianLeptogorgia virgulata reveals that spicule formation begins with the aggregation of scleroblasts in the mesoglea. Calcite crystal deposition occurs within a Golgi vacuole containing organic matrix. Vacuole size increases while matrix incorporation and subsequent crystal growth continue, filling the vacuole. At approximately this time, the scleroblasts dissociate and “wart” formation begins. Further spicule growth stretches the cell into a thin envelope. Fusion of vacuole and plasma membrane followed by breach formation during spicule growth, as well as scleroblast atrophy or migration from mature spicules, result in the transition of the spicule from the intracellular to the extracellular environment. The results also reveal aborted spicules and digestive bodies, implying possible relationships among calcification, detoxification, and waste management.
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  • 38
    ISSN: 1432-0827
    Keywords: Calcification ; Calciphylaxis ; Skin ; Serine Phosphate ; Threonine Phosphate ; γ-Carboxyglutamic Acid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The amount of non-collagenous proteins is increased greatly during the pathological calcification of rat skin experimentally induced by dihydrotachysterol (DHT) and Ovalbumin (topical cutaneous calciphylaxis). This is accompanied by an increase in the total amount and concentrations of protein-bound serine phosphate [Ser(P)], threonine phosphate [Thr(P)] and γ-carboxyglutamic acid (Gla), almost all of which can be extracted from the tissue and can be dissociated from collagen in 0.5M EDTA. The EDTA-soluble, non-collagenous proteins are rich in aspartic and glutamic acids, similar to the non-collagenous, EDTA-soluble proteins of bone, cementum and calcified cartilage, and quite distinct from those of dentin and enamel.
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  • 39
    ISSN: 1432-0878
    Keywords: Matrix vesicles ; Calcification ; Articular cartilage ; Synovialgraft ; Rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Free autologous grafts of synovial tissue were transplanted into experimental defects produced in the articular cartilage of rabbit knee joints. The grafted tissue underwent transformation into fibrocartilage. Extracellular matrix vesicles associated with calcified areas were present at the grafted sites. Hydroxyapatite crystals were found within these vesicles and in their vicinity. No calcification occurred in articular cartilage from sham operated joints in which defects were produced but no grafts made and in normal controls. These tissues showed abundant matrix vesicles devoid of crystalline mineral. A careful study of normal synovial tissue did not reveal matrix vesicles and calcifications. The present observations suggest that matrix vesicles in normal articular cartilage exist in a latent form. Vesicle mineralization following surgical manipulations of the joint is probably a manifestation of the metabolic stage of the tissue.
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  • 40
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    Calcified tissue international 32 (1980), S. 221-228 
    ISSN: 1432-0827
    Keywords: Medullary bone ; Calcification ; Low-calcium diet ; Parathyroid hormone ; Estrogens
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Medullary bone of birds maintained on a low-calcium diet represents a good model to study modifications of matrix composition in calcified tissue undergoing intense formation and resorption. The composition of the bone matrix during the low-calcium diet has been analyzed by both chemical and histological techniques. Sixty White Leghorn pullets 1 year old were used for the experiment. Fifteen birds served as controls and were killed on day zero; the remaining birds were placed on a calcium-deficient diet (0.13% calcium) and sacrificed after 4, 7, and 12 days of treatment in groups of 15. Serum levels of calcium, PTH, and estrogens were also measured. Chemical analysis of the samples were made for total nitrogen, hydroxyproline, hexosamine, hexoses, calcium, and phosphorus. Collagen and proteoglycans of the matrix of medullary bone of the egg-laying hens were found to be affected by the low-calcium diet. They either increased or decreased during the experiment but never in parallel. The increment of serum PTH is considered responsible for the variations in the amount of collagen. The effects of this hormone are magnified by the fall of serum estrogens as shown also by variations in the amounts of noncollagenous protein. In the late phase of the diet the matrix is represented by poorly calcified osteoid tissue rich in noncollagenous protein, i.e., proteoglycans and glycoproteins.
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  • 41
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    Calcified tissue international 30 (1980), S. 227-232 
    ISSN: 1432-0827
    Keywords: Matrix vesicles ; Epiphyseal cartilage ; Calcification ; Alkaline phosphatase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Matrix vesicles isolated from chicken epiphyseal cartilage displayed an uptake of Ca2+ which was linear with time and the amount of vesicle protein. The matrix vesicles stimulated the incorporation of Ca2+ even at very low Ca × P, suggesting that they could bind Ca2+ and/or increase the local Ca × P to the metastable level. This uptake was abolished by EDTA or heating, and partially inhibited by cysteine, to the same extent as the hydrolysis of ATP. There was also a certain uptake of Ca2+ without added phosphate, this being stimulated by ATP up to 3 mmol, but diminishing again with higher concentrations. The presence of ATP failed to stimulate the uptake of Ca2+ more than an equimolar amount of phosphate in the form of inorganic KH2PO4. Mg2+ activated the hydrolysis ofp-nitrophenylphosphate at pH 10.5, and both Mg2+ and Ca2+ the hydrolysis of ATP from pH 7 to 9.5. Paradoxically, the omission of Mg2+ stimulated the uptake of Ca2+ several-fold.
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  • 42
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    Calcified tissue international 30 (1980), S. 167-174 
    ISSN: 1432-0827
    Keywords: Proteolipid ; Calcium-phospholipid-phosphate complexes ; Calcification ; Hydroxyapatite ; Membranes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Calcium-phospholipid-phosphate complexes (Ca-PL-P) were isolated from calcified and uncalcifiedBacterionema matruchotii and its calcified lipid extracts. Similar complexes were absent from the noncalcifying bacteriumActinomyces naeslundii. The majority of the Ca-PL-P complexes were associated with the proteolipid acidic phospholipid component. Ca-PL-P complexes isolated fromB. matruchotii and from calcified proteolipid contained phosphatidylinositol, phosphatidylinositol-4-phosphate, phosphatidylinositol-4,5-diphosphate, and phosphatidylserine. They consisted of approximately 52 mole % Ca, 32 mole % organic P, and 15 mole % Pi. During Ca-PL-P extraction fromB. matruchotii or its proteolipid-containing calcified lipid extracts, the proteolipid was dissociated and the apoprotein precipitated as fluff at the aqueous-organic solvent interface, thus explaining the failure to detect protein in Ca-PL-P preparations. When the ability of Ca-PL-P complexes and lipid fractions ofB. matruchotii to initiate apatite formation from metastable calcium phosphate solution was compared, the yield of hydroxyapatite decreased as follows: Ca-PL-P 〉 proteolipid acidic phospholipids 〉 proteolipid 〉 crude phospholipid 〉 total lipids 〉 whole cells.
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  • 43
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    Calcified tissue international 32 (1980), S. 139-143 
    ISSN: 1432-0827
    Keywords: Calcification ; Magnesium ; Phospholipids ; Ca-phospholipid-phosphate complex ; hydroxyapatite
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The effect of Mg on hydroxyapatite proliferation induced by phosphatidyl serine, phosphatidyl inositol, and calcium-acidic phospholipidphosphate complexes has been studied in metastable calcium phosphate solutions of constant ionic strength and variable Mg/Ca ratio. Mg inhibits formation of the Ca-acidic phospholipid phosphate complexes, probably by competing with Ca for sites on the phospholipid molecules. Once the complexed acidic phospholipids are present, Mg has no effect on the proliferation of hydroxyapatite. This is shown by the invariant first-order rate constant for the disappearance of Ca during hydroxyapatite proliferation (kCa=0.0037 h−1) in solutions with Mg/Ca weight ratios ranging 0/1 to 10/1. These studies suggest that the presence of Mg does affect in vivo calcification and that the initiation of calcification by means of a Ca-PL-PO4 complex may be dependent on the Mg/Ca ratio in the calcifying tissue.
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  • 44
    ISSN: 1432-0878
    Keywords: Calcergy ; Calcification ; Cutaneous calcinosis ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In rats a subcutaneous injection of KMnO4 leads to the calcification of the connective tissue. During mineral deposition, both intra- and extracellular changes are observed in the connective tissue. The intracellular phase is characterized by the formation of intramitochondrial granules and cytoplasmic vesicles, both in fibroblastic and extrinsic cells. In the extracellular phase, numerous heterogeneous matrix vesicles appear in the extracellular matrix. At the same time, globular particles which are resistant to microincineration, are observed between the collagen fibrils. The mineralization of the extracellular matrix takes place in two stages. The first stage comprises the appearance of needle-like structures and round aggregates. The needle-like structures are observed occasionally in the matrix vesicles and often in the extracellular matrix where they appear isolated or diverging from a central point. The round aggregates, composed of dense particles, are seen in the ground substance between the collagen fibrils. The second stage is characterized by a progressive mineralization of the collagen fibrils and the elastic fibers, without formation of extended calcified plaques.
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  • 45
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    Cell & tissue research 211 (1980), S. 303-316 
    ISSN: 1432-0878
    Keywords: Fish scale ; Fine structure ; Development ; Calcification ; Cyprinodon variegatus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Scale formation in Cyprinodon variegatus was found to be initiated at about 26 to 30 days after hatching. Ultrastructural investigation revealed that within 4 to 6 h in the first-formed scales the marginal cells begin to flatten and differentiate into osteogenic cells, which later change to osteoblasts and fibroblasts. These cells are separated from the surrounding epithelial cells by a basal lamina. The osteoid is formed by the marginal and osteogenic cells; the osseous layer by the osteoblasts; and the fibrillary plate by the fibroblasts. The osteoid is formed within 2 to 3 h after the initiation of the scale, and within 20 to 24 h the osseous layer is formed. Hydroxyapatite crystals are deposited in the matrix of the osseous layer without apparent association with collagen fibers. No matrix vesicles or dense bodies are evident at the sites of calcification. The fibrillary plate arises 18 to 20 h after the initiation of the scale. It is also partially calcified, but not before the third week of scale formation. The crystals develop almost exclusively between the collagen fibers at the extreme edge of the calcifying front, but solid calcification of the fibers results with further growth of the crystals. The fibroblasts appear to participate in calcification of the fibrillary plate.
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    Medical & biological engineering & computing 18 (1980), S. 521-526 
    ISSN: 1741-0444
    Keywords: Artificial heart ; Biodegradation ; Biomaterials ; Blood compatibility tests in vitro and in vivo ; Calcification ; Protein adsorption ; Toxicity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Synthetic materials used for medical prostheses and devices, mainly those which will come into contact with blood, have to provide for specific properties. Three in vitro blood-compatibility tests are presented; one is a screening method (m.t.e.g.) whose results show remarkable correlations to in vivo behaviour, the other two (Nosé-blood-chamber, protein adsorption) yield more basis information on synthetic surfaces. Aspects of toxicity and biodegradation should be included in the definition of ‘blood compatibility’. After preselection by in vitro tests, a material has to pass an in vivo examination in animal experiments (shunt, catheter, tube, test chamber) under haemodynamic conditions. Finally, devices or prostheses in their intended shape, such as the total artificial heart, are implanted in animals.
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    Calcified tissue international 27 (1979), S. 75-82 
    ISSN: 1432-0827
    Keywords: Proteoglycans ; Hydroxyapatite ; Amorphous calcium phosphate ; Nucleation ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Well-characterized bovine nasal proteoglycan A1 fraction (aggregate) and proteoglycan D1 fraction (subunit) have been shown to be effective inhibitors of hydroxyapatite (HA) formation in two in vitro test systems: (a) the transformation of amorphous calcium phosphate (ACP) to crystalline HA, and, (b) the direct precipitation of HA from low-concentration calcium phosphate solutions. A1 or D1 in solution slowed the transformation kinetics in system (a) without affecting the time to the onset of conversion. In system (b), A1 or D1 in solution increased the time to the onset of HA formation without affecting the HA formation kinetics. In both test systems A1 was a more effective inhibitor than D1, although the difference was not great. In both systems the inhibitory effect was proportional to the A1 or D1 solution concentration. The action of solutions of low and high molecular weight neutral dextrans on both test systems showed that high molecular weight and/or extended spatial molecular conformation has a much stronger correlation with inhibitory ability than solution viscosity. Proteoglycans have been implicated as playing a role in regulating biological mineralization particularly in the epiphyseal growth plate. Our study suggests that just enzymatic cleavage of aggregate into subunit is not sufficient to allow mineralization to occur, since we find that D1 itself is a potent inhibitor of HA formation. Further degradation and/or removal of D1 appears to be necessary for calcification to take place.
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    Calcified tissue international 27 (1979), S. 83-88 
    ISSN: 1432-0827
    Keywords: Calcification ; Calcium-phospholipid-phosphate complexes ; Diphosphonates
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    Topics: Biology , Medicine , Physics
    Notes: Summary The diphosphonates disodium ethane-1-hydroxy-1, 1-diphosphonate (EHDP) and disodium dichloromethylene diphosphonate (Cl2MDP) prevent hydroxyapatite (HA) formation in metastable calcium phosphate solutions, induced by calcium-phospholipid-phosphate complexes and by the acidic phospholipids phosphatidyl serine and phosphatidyl inositol. The diphosphonates appear to act not only as HA crystal poisons but also as surfactants which probably change the nature of the lipid micelle and the charge and conformational properties of the lipid molecules. The surfactants sodium dodecyl sulfate (SDS) and Non-Idet P-40 (NP-40), like the diphosphonates, prevent HA formation by the acidic phospholipids and complexed lipids, but do not act as HA surface poisons. The lipid surfactant lyso-phosphatidyl serine did not induce HA formation from solution. The relevance of the ability of the diphosphonates to act as lipid surfactants to the in vivo use of these agents is discussed.
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    Calcified tissue international 29 (1979), S. 163-167 
    ISSN: 1432-0827
    Keywords: Mineralization ; Mollusks ; Organic matrix ; Calcification
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    Topics: Biology , Medicine , Physics
    Notes: Summary DEAE-cellulose ion-exchange chromatography separates soluble organic matrix components of three mollusk shells, each from a different taxonomic class, into analogous subfractions. The proteins of all subfractions are enriched in acidic and polar amino acids. In each chromatogram, however, the subfraction which contains the major portion of total protein also contains the highest concentration of aspartic acid. Thus the major components of the soluble organic matrix are aspartic acid-rich proteins. The identification of these proteins in mollusks, together with the known occurrence of aspartic acid and phosphoserine-rich proteins in vertebrate tooth dentin, emphasizes the fundamental importance of such acidic proteins in the processes of mineralization.
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    Calcified tissue international 29 (1979), S. 251-256 
    ISSN: 1432-0827
    Keywords: Cathepsin ; Calcification ; Dentinogenesis ; Proteoglycans
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    Topics: Biology , Medicine , Physics
    Notes: Summary Cathepsin D was purified from rat liver using a new affinity chromatographic method, based on the coupling to the specific inhibitor pepstatin. This preparation was used for the production of specific antibodies from rabbit. The purified IgG fraction was conjugated to horseradish peroxidase in a two-step coupling procedure and used for electron microscopic immunohistochemistry of the odontoblast-predentine region of the rat incisor. Precipitates, indicating the presence of cathepsin D, were seen in the odontoblast, odontoblast process, and in the extracellular unmineralized matrix, the predentine. The observations are discussed in relation to proteoglycan degradation at the mineralization front simultaneous with crystal formation, and in relation to the function of lysosomal enzymes in the turnover of connective tissue.
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    Calcified tissue international 29 (1979), S. 27-32 
    ISSN: 1432-0827
    Keywords: Ca2+-binding glycoprotein ; Pyrophosphatase ; ATPase ; Transphosphorylase ; Calcification
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    Topics: Biology , Medicine , Physics
    Notes: Summary The Ca2+-binding glycoprotein isolated from preosseous cartilage shows also alkaline phosphatase activity. The purification procedure indicates that the enzyme is inhibited in crude extract and conceivably in the intact tissue; the activity may be controlled by the proteoglycans present in the matrix. Other substrates are hydrolyzed by the purified enzyme in addition top-nitrophenylphosphate; the highest specific activity was measured with ATP and pyrophosphate (PPi) at pH 7.5 and 9.0 Mg2+ induces an activation of ATP and PPi hydrolysis; Ca2+ activates hydrolysis of ATP but inhibits that of PPi. The glycoprotein shows also transphosphorylase activity,l-serine being the best phosphate acceptor. The release or transfer of Pi catalyzed by the glycoprotein can be an important step in calcium phosphate precipitation.
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    Calcified tissue international 29 (1979), S. 101-105 
    ISSN: 1432-0827
    Keywords: Osteon ; X-ray diffraction ; Pole figures ; Electron microscopy ; Calcification
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    Topics: Biology , Medicine , Physics
    Notes: Summary The X-ray diffraction method based on pole figures has been applied to single osteon samples in order to obtain information about the texture of the inorganic bone fraction and the way it changes during calcification. The osteon samples were cylindrically shaped, with axes corresponding to those of the haversian canals. Selection was carried out according to the degree of calcification and the orientation of collagen bundles and inorganic particles. Osteons at both the initial and final stages of calcification were chosen. Arrangements of fiber bundles and inorganic particles in successive lamellae characteristic of three types of osteons were selected: longitudinal, alternate, and transversal. The results indicate that in all three types of osteons, the long axis of the sample is apparently the only direction of orientation because the transversally oriented crystallites give an isotropic diffuse scattering as would be expected if all the inorganic particles were irregularly oriented around the osteon axis. The number of longitudinally oriented crystallites increases progressively from transversally oriented osteons to alternately and longitudinally oriented ones. The crystallite orientation in an axial direction increases in fully calcified osteons. This last result is in agreement with the electron microscopic finding that the long needle-shaped crystallites covering much more than a major collagen period and measuring 40–45 Å in width increase in number as calcification proceeds.
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    Calcified tissue international 27 (1979), S. 57-64 
    ISSN: 1432-0827
    Keywords: Bone ; Bone formation ; Calcification ; Calcification nodule ; EM
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    Topics: Biology , Medicine , Physics
    Notes: Summary Osteolathyrism has been used as an experimental model for the study of calcification nodules during the mineralization process. Periosteal exostoses developing in osteolathyrism characteristically have spherical basophilic structures (calcification nodules) in the vicinity of developing bone spicules. In thin sections, the nodules were seen scattered between collagen fibers in the intercellular matrix. Collagen fibers did not appear to be present within the nodules but sometimes were packed just outside them. Matrix vesicles were also present in areas of early mineralization. After EDTA decalcification, the majority of the nodules consisted of a fine granular material surrounded by an electron-dense peripheral zone. The peripheral dense zone was occasionally incomplete in small nodules in areas of early mineralization. An electron-dense central area could be observed in the center of the nodules. Evidence has been presented indicating that the calcification nodules arise from smaller mineralization foci, presumably matrix vesicles. The calcification nodules enlarge to approximately 1.0 µm in size, at which point development is slowed or halted allowing the formation of the peripheral dense zone. Although coalescence of nodules was observed, this was more a random event. The further mineralization of the trabeculae was achieved by the calcification of the collagen fibers. The mineralized trabeculae reflected this pattern of nodular and collagenous calcification. It is suggested that this pattern of calcification is characteristic of rapidly developing woven bone.
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    Calcified tissue international 27 (1979), S. 177-185 
    ISSN: 1432-0827
    Keywords: Calcification ; Bone ; Glycoprotein ; Golgi ; Osteoblasts
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    Topics: Biology , Medicine , Physics
    Notes: Summary The elaboration of bone matrix glycoprotein by osteoblasts of alveolar bone was investigated by radioautography after the intravenous injection of3H-fucose into young rats. At selected times after injection, animals were sacrificed by intracardiac perfusion and demineralized specimens were prepared for light and electron microscope radioautography. At 5 and 10 min after injection, when the blood fucose level was high, silver grains were restricted to the spheroidal and cylindrical saccules of the Golgi apparatus. At 20 min membrane-limited secretory granules were also labeled. By 35 min, the blood fucose level had dropped and silver grains were detected over the apical cortical cytoplasm, in association with secretory granules located therein. Some grains were present over osteoblast processes and the adjacent prebone matrix. By 4 h most of the silver grains had left the cell. At that time they were observed over prebone, adjacent to osteoblast processes, as well as over the prebone-bone junction where a distinct band of label was noted. In demineralized preparations an electron-dense granular material was present at the prebone-bone junction in association with collagen fibrils. These findings provide evidence that osteoblasts in alveolar bone synthesize fucose-containing glycoprotein and indicate that the addition of3H-fucose occurs in the Golgi apparatus. The glycoprotein passes to the apical cortical cytoplasm by way of membrane-limited secretory granules, is exteriorized, and accumulates at the site where prebone transforms into bone (the prebone-bone junction). Since this is also the site of the calcification front, the deposition of labeled glycoprotein may be related to the deposition of bone mineral.
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    Plant systematics and evolution 131 (1979), S. 169-178 
    ISSN: 1615-6110
    Keywords: Cyanophyceae ; Cyanobacteria ; Stigonemataceae ; Geitleria calcarea Friedmann ; Geitleria floridana Friedmann ; n. sp. ; Calcification ; fine structure of sheath ; scanning electron microscopy ; cave algae ; atmophytic ; nomenclature
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    Notes: Abstract The calcifying cave inhabitant atmophytic blue-green algaGeitleria calcarea is reported from new localities in Florida and in the Cook Islands.—G. floridana n. sp., is described from caves in Florida. The calcified sheath has the shape of a quadratic prism and is built of crystalline acicular subunits about 0.1 µm in diameter. The subunits mostly form a rhombic lattice pattern but in some cases, they are not distinguishable and then the surface of the sheath is smooth.
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    Cell & tissue research 201 (1979), S. 409-422 
    ISSN: 1432-0878
    Keywords: Fish scale ; Fine structure ; Calcification ; Fibrillary plate ; Carassius auratus
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    Topics: Biology , Medicine
    Notes: Summary Electron microscopic investigation of scales of the goldfish Carassius auratus revealed that the lamellae of fibrillary plates contain sheet-like structures composed of vertically oriented collagen fibers embedded in an organic matrix. The fibers (TC fibers) are smaller in diameter (35–45 nm) than those of the lamellae and the matrix is stained intensely with lead citrate. The sheet-like structures as well as the lamellae are formed by fibroblasts located beneath the lamellae. The orientation of the collagen fibers of the sheets and the lamellae seems to be controlled by the orientation of the ridges and invaginations of the surface of the fibroblasts. The fibrillary plate of C. auratus was found to be partially calcified. Calcification was initiated by the deposition of needle-like or flaky crystals of hydroxyapatite in the organic matrix of the sheet-like structure and proceeded into the TC fibers and the matrix region of the lamellae. The potassium pyroantimonate-osmium tetroxide method showed a heavy concentration of calcium in the osteoblasts, fibroblasts, and in the matrix regions of the fibrillary plate. Calcium-containing precipitates were also present in the “hole zone” of the collagen fibers in the lamellae, but the significance of this location in calcification remains to be elucidated.
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    Calcified tissue international 26 (1978), S. 51-59 
    ISSN: 1432-0827
    Keywords: Collagen ; Calcification ; Fluoroapatite ; Nucleation ; Inhibitors
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    Topics: Biology , Medicine , Physics
    Notes: Summary Ca2+ and Pi uptake induced in vitro by a collagenous matrix derived from bovine tendon is inhibited by 1×10−6 to 2×10−5 M NaF and stimulated by 2×10−5 to 2×10−3 M NaF. Fluoride uptake occurs only over the latter concentrtion range. The uptake of Ca2+, Pi, and F−1 progresses toward a limiting extent at which the molar Ca/P and Ca/F values are 1.6 to 1.7 and 4.5 to 5.7, respectively. Although the matrix-bound mineral, previously formed in the absence of NaF, readily undergoes dissolution when exposed to a Ca2+- and P-free medium of pH〈7.4, the bound mineral phase formed in the presence of NaF does not. We conclude that fluoroapatite is the primary matrix-bound mineral. The uptake of fluoride, Ca2+. and Pi by both uncalcified and previously calcified matrices is inhibited by methylenediphosphonate and by phosphonoacetate as is calcification in the absence of NaF. Kinetic studies indicate that formation of a CaP complex precedes the uptake of F−1 and suggest that F−1 and OH−1 compete for interation with the CaP complex during the calcification process. We concluded that fluoroapatite formation induced by the collagenous matrix occurs by a multistep pathway comparable to that proposed previously for hydroxyapatite formation.
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    Calcified tissue international 26 (1978), S. 253-258 
    ISSN: 1432-0827
    Keywords: Ion-exchange resin ; Lead toxicity ; Energy dispersive X-ray analysis ; Calcification ; Trauma
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    Topics: Biology , Medicine , Physics
    Notes: Summary Several heavy metals have direct calcifying effects in connective tissues; most notable among them is lead (Pb), whether administered topically (1) or systemically with local injury (2). The mechanisms of metal-induced soft tissue calcification have been studied by injecting salts of known in vitro calcifying potential into subcutaneous connective tissue (3); of the metals used, only lead and holmium produced an early accumulation of minerals on collagen in vivo. Lead is also claimed to accelerate bone healing in the rabbit leg with no toxic effects (4). Lead-loaded ion-exchange resin beads, implanted into surgically prepared subcutaneous pouches in rats, rapidly induced subcutaneous calcification which has been studied by microradiography and electron-probe microanalysis.
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    Calcified tissue international 25 (1978), S. 145-159 
    ISSN: 1432-0827
    Keywords: Bird egg shell ; Ultrastructure ; Calcification ; Electron diffraction ; Microanalysis
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    Topics: Biology , Medicine , Physics
    Notes: Summary The egg-shell of Japanese quail was studied by several techniques. Semithin sections (1μm thick) of non-decalcified shell were observed by normal and polarized light microscopy. Thin sections of non-decalcified shell, examined by transmission electron microscopy, permitted us to observe the forms and dimensions of crystals of calcite within different layers of the shell: mammilary layer, layer of cones, palissade layer and surface crystal layer. There appears to be two distinct zones in the layer of cones as well as in the superficial crystal layer. Electron microdiffraction revealed the orientation of calcite crystals in the columns. Some crystal defects (twins?) were described and the possibility of their artefactual formation during ultramicrotomy is discussed. Localization of Ca, Mg, P and S were made by X-ray microanalysis of semithin sections. This technique shows that shell membranes, and chiefly the true cuticle, are also mineralized but, in these layers, minerals are not crystallized. Otherwise the distribution of Mg is not uniform throughout the shell thickness; it is less concentrated in the external zone of the layer of cones. These results together with observation of developing shells by scanning electron microscopy allowed us to propose a scheme for shell organization of the quail egg. This organization was related with decalcification which occurs during hatching.
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    Calcified tissue international 25 (1978), S. 133-143 
    ISSN: 1432-0827
    Keywords: Osteon ; X-Ray diffraction ; Electron microscopy ; Calcification
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    Topics: Biology , Medicine , Physics
    Notes: Summary To obtain information on the changes in the inorganic bone fraction during calcification, low- and wide-angle X-ray diffraction techniques and electron microscopy have been applied to single osteon samples. The samples were cylindrically shaped and their axes corresponded to the axes of the Haversian canals. The selection was made according to the degree of calcification and the orientation of collagen bundles and inorganic particles. Osteons at both the initial and final stages of calcification were chosen. Arrangements of fiber bundles and inorganic particles in successive lamellae characteristic of three types of osteon were selected, that is, longitudinally structured osteons, transversely structured osteons, and alternately structured osteons. The results indicate that in osteonic lamellar bone there are two types of inorganic particles: (1) granules arranged in linear or needle-shaped entities with maximum width 40–45 Å, which are regularly distributed at the level of the main band of the collagen fibrils where their maximum length reaches the length of the main band itself; that is, about 400 Å; and (2) very long crystallites, with a diameter of 40–45 Å, which grow with their crystallographicc-axis parallel to the collagen fibrils and cover much more than a major collagen period.
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    Calcified tissue international 26 (1978), S. 267-269 
    ISSN: 1432-0827
    Keywords: Chondroid tissue ; Calcification ; Opilions ; Chelicerates ; Electron microprobe
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    Notes: Summary Extracellular masses of crystals are present in the endosternite of certain opilions (Phalangium, Odiellus, andLeiobunum). X-ray spectrography and secondary-ion analysis have shown them to be rich in calcium but poor in phosphorus and other elements. The associated anion has not been identified, but is most probably organic in nature and perhaps an oxalate.
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    Calcified tissue international 25 (1978), S. 161-168 
    ISSN: 1432-0827
    Keywords: Tendon ; Calcification ; Collagen ; Inhibitors ; Proteoglycans
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    Topics: Biology , Medicine , Physics
    Notes: Summary Bovine and human tendon tissue do not induce calcification in vitro. However, extraction of those tissues with 3% Na2HPO4 converts them to calcifiable matrices. The supernatant fraction derived from the extraction contains a nondialyzable, perchloric acid soluble component that inhibits calcification of the extracted matrix. This inhibitory substance is characterized by a molecular weight in the range of 85,000–100,000. Exposure to pronase or hyaluronidase did not alter the inhibitory potency but did render the inhibitor dialyzable. Commercial sources of hyaluronic acid, chondroitin-6-sulfate, chrondroitin-4-sulfate, dermatan sulfate, heparin and lysozyme did not inhibit calcification of the extracted matrix. Phosvitin, a phosphoglycoprotein is a potent inhibitor. Although phosvitin and the tendon extract also inhibit calcification of previously calcified matrix, they have no detectable effect on the rate of decalcification. We conclude that the mechanism of inhibition is characterized by a degree of specificity and that phosvitin and a macromolecular component of tendon tissue blocks conversion of an intermediate matrix-bound CaP complex to crystalline apatite. It seems reasonable that the tendon inhibitor could function in situ and possibly in vivo to control calcification of tendon tissue.
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    Cell & tissue research 192 (1978), S. 249-266 
    ISSN: 1432-0878
    Keywords: Calcification ; Calcite ; Collagen ; Pennatulid
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    Topics: Biology , Medicine
    Description / Table of Contents: Résumé L'axe squelettique de Veretillum cynomorium se compose d'une matrice fibrillaire de nature collagène, minéralisée par de la calcite. Le présent article fournit des données ultrastructurales et cristallographiques quant à l'organisation de cette structure et au dépôt du minéral dans la matrice. La pointe inférieure de l'axe constitue un gradient de calcification entre l'extrémité non minéralisée et le corps entièrement calcifié. Les dépôts initiaux de calcite, parfois associés à des débris cellulaires, donnent naissance à des nodules qui s'accroissent de façon rayonnée, par bourgeonnement de plusieurs lobes. Ces nodules fusionnent et forment le coeur de l'axe dont la croissance ultérieure en diamètre, se réalise par le développement de colonnes irrégulières de calcite à partir des nodules externes du coeur. Les surfaces minéralisées présentent une microarchitecture qui peut être reliée aux axes cristallographiques de la calcite. Les relations entre le minéral et la matrice ont particulièrement retenues notre attention. Les fibres de collagène, enrobées par la calcite et non imprégnées par elle, n'interviennent jamais comme initiateur de nucléation du minéral. L'orientation cristallographique ultérieure de la calcite est aussi totalement indépendante de la matrice.
    Notes: Summary The axial skeletal rod of Veretillum cynomorium consists of a fibrillar collagenous matrix calcified with calcite. The present paper describes ultrastructural and crystallographic details of its organization and deposition. At the inferior end of the rod is a calcification gradient between the noncalcified tip and the rest of the axis. Initial mineral deposits, which are sometimes associated with cell debris, give rise to calcitic nodules which enlarge by the radial growth of several lobes. These nodules fuse and form the core of the axis. Subsequent increase in diameter of the rod involves the radial development of irregular columns of calcite which arise from the peripheral nodules. Mineral surfaces exhibit a distinctive microarchitecture which can be related to the predominantly c-axis parallel growth of the calcite. Particular attention is paid to the relationship between mineral and matrix. The collagen fibrils, embedded in the calcite but never impregnated with it, are not responsible for the initial nucleation of mineral. The crystallographic orientation of the calcite also appears to be independent of these fibrils.
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    Calcified tissue international 22 (1977), S. 159-172 
    ISSN: 1432-0827
    Keywords: Calcification ; Artery ; Gastropod ; Spherule
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    Notes: Abstract The anterior aorta and major arteries of the land pulmonate snailAnguispira alternata have large calcium deposits in their walls. These deposits occur inside spherule cells, which line the walls of these vessels. The calcium occurs as amorphous calcium carbonate, in the form of intracellular spherules having alternating layers of organic and inorganic material. The spherule cells appear to be degenerating connective tissue cells; they are characterized by large numbers of spherules and by a cytoplasm which is totally empty except for a nucleus, scattered glycogen particles and a few membrane remnants. The injection of45Ca into the foot of the snail results in rapid incorporation of this calcium into the spherule cells. Although calcium-containing spherules are now known from a wide variety of invertebrate tissues, they have not been previously recorded from arterial walls. The physiologic significance of these deposits is not known.
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    Calcified tissue international 23 (1977), S. 33-38 
    ISSN: 1432-0827
    Keywords: Cathepsind ; Calcification ; Odontoblasts ; Dentinogenesis
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    Notes: Summary The presence of an acid proteinase with a high activity has been demonstrated in isolated odontoblast-predentine material from dentinogenically active rat incisors. The enzyme was identified as cathepsind (EC 3.4.23.5). The possible significance of the enzymatic degradation of proteoglycans and glycosaminoglycans in the course of the calcification process is discussed.
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    Calcified tissue international 22 (1977), S. 1-7 
    ISSN: 1432-0827
    Keywords: Matrix vesicles ; Alkaline phosphatase ; Pyrophosphatase ; ATPase ; Calcification
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    Notes: Abstract Some of the characteristics of the pyrophosphatase and ATPase activities studied in isolated cartilage matrix vesicles were found to be similar to those already reported for the solubilized and purified bone alkaline phosphatase. Thus, the pH optimum of the pyrophosphatase activity responded similarly to changes in the concentration of Mg2+, Ca2+, and PPi. Further, the ATPase activity was not activated by Ca2+ in the presence of an optimal Mg2+ concentration. It is proposed that a function of the alkaline phosphatase of matrix vesicles in vivo is to hydrolyze the substrates PPi, ADP, and ATP, which are known inhibitors of calcium phosphate precipitation.
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    Calcified tissue international 22 (1977), S. 129-135 
    ISSN: 1432-0827
    Keywords: Calcification ; Connective tissue ; Skeletal muscle
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    Notes: Abstract The subcutaneous injection into mice of small quantities of lead salts resulted in calcification of the dorsal fascia. Other reputed calcergens failed to produce a similar reaction. However, the injection of Zn Cl2 and KMn O4 in high doses caused damage to, and subsequent calcification of the panniculus carnosus muscle.
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    Calcified tissue international 22 (1977), S. 231-241 
    ISSN: 1432-0827
    Keywords: Alkaline phosphatase ; Calcification ; Cartilage ; Enamel organ ; Odontoblasts
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    Notes: Abstract The enzyme alkaline phosphatase (AP) (EC 3.1.3.1) in three different calcification areas was studied by means of a spectrophotometric micro method using p-nitrophenylphosphate as a substrate. Rat maxillary incisor odontoblasts and enamel organ from the zones of matrix formation and maturation and tissue from rabbit metatarsal cartilage were allowed to react with the substrate in glycine-NaOH buffer at room temperature. The reaction was found to be linear for a minimum of 20 min. The pH optima for AP from these tissues were in the pH range of 10.0–10.3. In order to compare AP from the four calcification areas different parameters were studied. Heating at 56°C or 60°C for varying times revealed that the enzymes were almost completely inactivated after 10 min. Mg2+ ions activated the enzymes by about 25% at concentrations of 2.5 mM (enamel organ 1.25 mM); while only higher concentrations of Mg2+ had an inactivating effect, Ca2+ and PO 4 3− ions were inactivating at varying concentrations. F− ions show no effect on AP activity at concentrations below 250 mM (enamel organ 125 mM) but caused inactivation of the enzymes at about 50% at 1 M. EDTA was found to be a very effective AP inactivator at concentrations above 0.06 mM, whereas urea did not noticeably affect the enzyme reactions at concentrations below 1 M. At higher concentrations, inactivation was observed. In order to determine AP localization in the epiphyseal plate successive 40-μm-thick, freeze-sectioned slices were analyzed. The activity was highest nearest the zone of cartilage calcification and decreased towards the reserve cell zone. It was concluded that the same AP isoenzyme is present in these quite different calcification loci.
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    Calcified tissue international 23 (1977), S. 259-269 
    ISSN: 1432-0827
    Keywords: Amorphous calcium phosphate ; Apatite ; Calcification ; Octacalcium phosphate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The maturation of calcium phosphate crystals formed by the conversion of spontaneously precipitated amorphous calcium phosphate (ACP) was studied in aqueous media at temperatures ranging from 20° to 37°. Reaction pH was kept at 7.4 with either Hepes buffer or by the pH-stat addition of base. Reaction kinetics were followed by monitoring solution calcium and total phosphate, and, in the pH-stat controlled reaction, by recording the amount of KOH needed to maintain pH. Reaction products were examined chemically and by X-ray diffraction and transmission electron microcopy. The first crystals to form deviated markedly from apatite in morphology, composition, structure, and solubility. They were extremely thin and flaky in appearance, had a low Ca/P molar ratio (1.4), contained an appreciable amount of acid phosphate (16%), and had an exceptionally largea-axis (10.5 Å vs. 9.4 Å for apatite). With maturation, the crystals became thicker but smaller in lateral dimensions, more apatitelike in structure and composition, and less soluble. However, this ripening of the crystals was accompanied by unusual inflections in the solution Ca and total PO4 curves, and, in the case of the pH-stat experiments, in the OH consumption profiles as well. These anomalous post-ACP solution changes suggest that a phase change had taken place during crystal maturation. Although the observed structural and compositional changes are not inconsistent with the perfection of an initially defective apatite, the changes in crystal morphology and the anomalous behavior of the reaction solution may more accurately reflect a conversion of the ACP first into an OCP-like crystalline phase which subsequently hydrolyzes into apatite.
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    Calcified tissue international 24 (1977), S. 47-57 
    ISSN: 1432-0827
    Keywords: Calcification ; Crystal growth ; Hydroxyapatite ; Kinetics ; Precipitation
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary The kinetics of precipitation of hydroxyapatite, HA, was studied by seeding dilute supersaturated solutions with well characterized HA crystals. In solutions having initial degrees of supersaturation comparable to those present in human serum, the precipitation rates were related to the thermodynamic driving force (degree of supersaturation with respect to HA) and not to the solution composition. The following relationshipR 0=KA(DS) n , whereR 0=initial precipitation rate, A=amount of seeds, DS=degree of supersaturation, and K andn are parameters obtained from the experimental data, was found to apply over a DS range of 6.6×1010 to 3.3×106. These observations are consistent with the occurrence of a simple growth process on the HA seeds. No evidence for the formation of discrete calcium phosphates other than HA was detected. The Ca/P molar ratio of the precipitating phase, calculated from solution compositions, was invariably higher than that expected for HA; this result is shown to be consistent with an initial adsorption phenomenon. Anomalous kinetic behavior was observed at low seeding levels and may relate to the surface phenomenon described. It is concluded that, most probably, under physiological conditions, formation and remineralization of hard tissues occur through the reported crystal growth process.
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    Cell & tissue research 167 (1976), S. 37-47 
    ISSN: 1432-0878
    Keywords: Scale ; Tilapia ; Calcification ; Ultrastructure
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    Topics: Biology , Medicine
    Notes: Summary The scales of Tilapia are surrounded by an envelope of scleroblasts responsible for the production of layers of collagen that constitute the bulk of the scale. The scleroblasts adjoining the lateral face of the oldest scale region gradually atrophy. New collagen layers are deposited against the inner face of the scale, the adjoining scleroblasts showing evidence of high metabolic activity. Calcification occurs by inotropic deposition of crystals alongside the fibres. There is no sharp demarcation between calcified and non-calcified scale regions, a calcification front gradually moving towards newly formed collagen layers. It is felt that fish scales should be considered as calcified derivatives of dermal collagen layers.
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    Cell & tissue research 173 (1976), S. 349-356 
    ISSN: 1432-0878
    Keywords: Finrays ; Tilapia ; Ultrastructure ; Calcification
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    Topics: Biology , Medicine
    Notes: Summary Light and electron-microscopic studies were carried out on the fins of the fish Tilapia mossambica (Peters). A detailed description is presented of the different skeletal components comprising the finrays. The mode of development of the hemisegments appears in several ways similar to that of fish scales. Each hemisegment is contained by an envelope of scleroblasts which secrete collagen fibres in a unipolar fashion. Calcification takes place as a result of deposition of hydroxyapatite-like crystals between the collagen fibres. However, the orientation of these fibres is not as regular as that of the fibres occurring in scales.
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    Cell & tissue research 168 (1976), S. 277-287 
    ISSN: 1432-0878
    Keywords: Proteoglycans ; Odontoblasts, predentin, dentin ; Calcification ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The localization of proteoglycans in the predentin of the rat incisor was investigated by ultrastructural histochemistry. Ruthenium red stained the cell coat of the odontoblasts as well as intracellular vesicles. There was also a staining of the extracellular matrix, but not of collagen fibers in the predentin. Treatment with the enzyme hyaluronidase prior to staining with ruthenium red abolished the staining of the vesicles and the extracellular matrix but not that of the cell coat. Bismuth nitrate and phosphotungstic acid gave similar staining of odontoblast vesicles and extracellular matrix. It is likely that the stained structures contain proteoglycans. The importance of these proteoglycans and their ultrastructural localization are discussed in relation to intracellular transport and the calcification process.
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    Cell & tissue research 163 (1975), S. 81-97 
    ISSN: 1432-0878
    Keywords: Medullary bone ; Bone ultrastructure ; Bone histochemistry ; Calcification ; Organic-inorganic relationships
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary Folliculin administration to pigeons stimulates the development of medullary bone in marrow spaces of the femora and other long bones. It is a specialized osseus tissue not devoted to mechanical functions and which is rapidly reabsorbed before egg-shell formation. Medullary bone is formed and reabsorbed in the same way as other types of bone. Consequently, because of its very rapid rate of formation and resorption, it represents an ideal tissue for studying osteoblastic, osteoclastic and osteocytic activity, and the calcification process. Medullary bone is deeply stained by PAS, Alcian blue and colloidal iron and is metachromatic after toluidine blue staining. This shows that its interfibrillary ground substance contains relatively high amounts of glycoproteins and acid proteoglycans. Calcification initially occurs in matrix vesicles (or calcifying globules) which are very numerous between the collagen fibrils of the osteoid tissue, and successively spreads into the surrounding interfibrillar matrix. Here, the crystals are closely related to thin, filament-like organic structures which seem to be components of ground substance proteoglycans. These findings confirm that in medullary bone, as in other types of calcifying tissue, the inorganic substance is initially laid down within calcifying globules and is successively closely related to organic, non-collagenous, filamentous organic structures (crystal ghosts) which probably represent a framework for calcium salt deposition.
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    Cell & tissue research 159 (1975), S. 233-243 
    ISSN: 1432-0878
    Keywords: Odontoblasts ; Predentine ; Dentine ; Calcification ; Scanning electron microscopy
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    Topics: Biology , Medicine
    Notes: Summary A scanning electron microscopic technique was used to investigate the surface structure of dentinogenically active odontoblasts. Thin pieces of rat incisors were fixed, rapidly frozen, freezedried at -70° C and fractured to expose new surfaces prior to examination in the SEM. Differences in the appearance of odontoblastic cell surfaces were seen, with the most extensive ridge formations at the distal part of the sides of the odontoblasts. The predentine area displayed a spongy structure which contrasted to the compact appearance of dentine. Results are discussed in relation to previous studies at the light microscopic and transmission electron microscopic levels.
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    Calcified tissue international 16 (1974), S. 227-237 
    ISSN: 1432-0827
    Keywords: Kinetics ; Calcification ; Secretion ; Protistan ; Coccolith
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    Notes: Abstract The calcified body scale (coccolith) of the unicellular marine algaHymenomonas carterae is composed of two morphologically distinct calcite elements which alternate about the perimeter of a complex, elliptical organic base. The coccoliths ofHymenomonas are formed intracellularly in a precise and orderly sequence of events within the single Golgi apparatus of the alga and then secreted to the anterior cell surface. This study demonstrates that the kinetics of this calcification-secretion process can be reproducibly determined by morphological means. Under standardized conditions, coccoliths are secreted at a rate of about ten per hour. In the absence of light this rate is significantly reduced. The procedure described for determining the kinetics of coccolith secretion inHymenomonas provides a simple, precise method for evaluating the results of experimental manipulation of this calcification-secretion system.
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    Calcified tissue international 16 (1974), S. 315-320 
    ISSN: 1432-0827
    Keywords: Ossification center ; Calcification ; Femoral head ; Estrogen ; Hypophysectomy
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    Topics: Biology , Medicine , Physics
    Notes: Abstract In Wistar rats, calcified centers normally appeared in the epiphyseal cartilage of the femoral heads by 21 days of age, but ossification centers were not observed until the animals were at least 6 weeks old. The effects of hypophysectomy, performed when the rats were 13 days old, and estradiol, 5 μg per g body weight injected subcutaneously on the day of birth and subsequently every second day, were investigated. All animals were sacrificed when 21 days old and the development of calcified centers examined radiographically and histologically. At this time there were fewer calcified centers in the femoral heads of the hypophysectomized rats than in those of the control animals, but there was no significant difference between the number of calcified centers in the estradiol-treated hypophysectomized rats and the controls. Estradiol-injected rats developed calcified centers at a nearlier stage than hypophysectomized rats receiving estradiol injection. These findings are interpreted to show that hypophysectomy produces the opposite effect to estradiol treatment on the development of calcified centers in the femoral heads of rats.
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    Calcified tissue international 14 (1974), S. 31-48 
    ISSN: 1432-0827
    Keywords: Bone ; Cartilage ; Calcification ; Collagen ; Phosphate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Une méthode pour l'étude de la croissance des os longs de foetus de rat, en culture d'organe, dans un milieu chimiquement défini, a été mise au point. Les extrémités cartilagineuses et les parties centrales de l'os sont analysées séparément pour leur croissance et minéralisation en étudiant leur contenu en collagène, calcium et phosphate, poids sec, et incorporation de proline marquée en hydroxyproline. La croissance et la minéralisation des parties centrales osseuses sont plus lentes dans un milieu chimiquement défini qu'in vivo. La croissance peut être accélérée en ajoutant au milieu des acides aminés non essentiels, de l'albumine ou du sérum. Les extrémités cartilagineuses présentent une augmentation plus importante en poids et contenu en collagène que les parties centrales et l'adjonction de diverses substances a moins d'effet sur la croissance. La croissance et la minéralisation des parties centrales sont augmentées en élevant la concentration du milieu en phosphate de 1.5 à 4.5 mM, avec ou sans adjonction de sérum ou d'albumine. A une concentration faible de calcium (0.5 mM), la croissance et la minéralisation des parties centrales sont arrêtées. A une concentration faible en magnésium (0.5 mM), la minéralisation est augmentée, mais la croissance est arrêtée.
    Abstract: Zusammenfassung Es wird eine Methode beschrieben, mit welcher das Wachstum der Röhrenknochen von Rattenembryos in einem chemisch bestimmten Medium in Organkultur untersucht werden kann. Die Knorpelenden und Knochenschäfte wurden gesondert auf Wachstum und Mineralisation geprüft, indem Collagen-, Calcium- und Phosphatgehalt, das Trockengewicht und der Einbau von markiertem Prolin in Hydroxyprolin gemessen wurden. Wachstum und Mineralisation des Knochenschaftes waren langsamer in einem chemisch bestimmten Medium als in vivo. Das Wachstum konnte beschleunigt werden, indem dem Medium nicht-essentielle Aminosäuren, Albumin oder Serum beigegeben wurden. Die Knorpelenden zeigten eine viel stärkere Zunahme an Gewicht und Collagengehalt als die Schäfte, und Anreicherung des Mediums hatte weniger Wirkung auf ihr Wachstum. Das Wachstum und die Mineralisation der Knochenschäfte nahmen zu, wenn die Phosphatkonzentration im Medium zwischen 1,5 und 4,5 mM erhöht wurde, und zwar unabhängig davon, ob dem Medium Serum oder Albumin beigegeben wurde oder nicht. Bei niederer Calciumkonzentration (0,5 mM) im Medium wurden Wachstum und Mineralisation der Knochenschäfte beeinträchtigt. Bei niedriger Magnesiumkonzentration (0,5 mM) wurden die Mineralisation erhöht, das Wachstum hingegen gehemmt.
    Notes: Abstract A method for studying the growth of fetal rat long bones in a chemically defined medium in organ culture is described. Cartilage ends and bone shafts were analyzed separately for growth and mineralization by measuring the collagen, calcium, and phosphate content, dry weight, and incorporation of labeled proline into hydroxyproline. Growth and mineralization of the bone shaft were slower in a chemically defined medium thanin vivo. Growth could be enhanced by supplementation of the medium with non-essential amino acids, albumin or serum. Cartilage ends showed a greater increase in weight and collagen content than the shafts, and medium supplements had less effect on their growth. Bone shaft growth and mineralization were enhanced by increasing medium phosphate concentration over a range of 1.5 to 4.5 mM whether or not the medium was supplemented with serum or albumin. At a low medium calcium concentration (0.5 mM) bone shaft growth and mineralization were impaired. At a low magnesium concentration (0.5 mM) mineralization was enhanced, but growth was impaired.
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    Calcified tissue international 14 (1974), S. 275-281 
    ISSN: 1432-0827
    Keywords: Calcification ; Cartilage ; Lipids ; Membranes ; Vesicles
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Les vésicules matricielles sont des particules extracellulaires, entourées par une membrane, qui sont nombreuses dans la matrice du cartilage en voie de calcification et qui sont en rapport morphologique avec le dépôt minéral. Les vésicules matricielles et les cellules de l'épiphyse d'os longs d'embryons de veaux sont libérées par digestion grâce à la collagénase et séparées par centrifugation différentielle. Les vésicules ainsi obtenues sont riches en lipides et contiennent significativement plus de sphingomyéline et phosphatidyl sérine que les fractions cellulaires. Le rapport moles de cholestérol sur moles de phospholipides totaux est plus élevé dans les vésicules que dans les cellules; ce résultat est en accord avec l'origine des vésicules au niveau des membranes cellulaires des chondrocytes.
    Abstract: Zusammenfassung Matrixvesikel sind extrazelluläre, membrangebundene Teilchen, welche in der Matrix von verkalkendem Knorpel gehäuft vorkommen und welche in morphologischem Zusammenhang mit der Mineralablagerung in diesem Gewebe stehen. Matrixvesikel und Epiphysenzellen aus Röhrenknochen von Kalbsembryonen wurden mittels Collagenaseverdauung freigesetzt und durch Differential-Zentrifugierung getrennt. Die Vesikelpräparate waren reich an Lipidien und enthielten bedeutend mehr Sphingomyelin und Phosphatidylserin als die Zellfraktionen. Das molare Verhältnis Cholesterol/Gesamtlipide war in den Vesikeln höher als in den Zellen; dieser Befund stimmt mit der Tatsache überein, daß die Vesikel ihren Ursprung in den Plasmamembranen der Chondrozyten haben.
    Notes: Abstract Matrix vesicles are extracellular, membrane-bounded particles which are abundant in the matrix of calcifying cartilage and which are morphologically related to the deposition of mineral in this tissue. The matrix vesicles and cells of the epiphyses of the long bones of fetal calves were liberated by digestion with collagenase and separated by differential centrifugation. Vesicle preparations were found to be lipid-rich and to contain significantly more sphingomyelin and phosphatidyl serine than cellular fractions. The ratio, moles cholesterol to moles total phospholipid, was higher in vesicles than in cells, a finding which is consistent with vesicles having their origins in the plasma membranes of the chondrocytes.
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    Calcified tissue international 14 (1974), S. 317-325 
    ISSN: 1432-0827
    Keywords: Calcification ; Neutral sites ; Elastin ; Calcium
    Source: Springer Online Journal Archives 1860-2000
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    Description / Table of Contents: Résumé Des études précédentes suggèrent la présence de groupements carboxyles, sulfhydriles et aminés dans les sites de liaison en calcium de l'élastine. La possibilité de l'existence de sites neutres de liaison en calcium au niveau de l'élastine a été étudiée dans ce travail. Une augmentation de la fixation du calcium au niveau de l'élastine est observée après des modifications de dissolution qui ont aussi provoqué des modifications de structure de la protéine. Dans des mélanges méthanol-H2O, les liaisons du calcium semblent indépendantes du pH et de la force ionique. Sur dix ions testés (Ca2+, CO2+, Na2+, Cu2+, Zn2+, Cr3+, Pb+2, K+, Rb+ et Mg2+) seule la liaison du calcium est nettement augmentée, lorsque le méthanol est ajouté. Il semble que les sites neutres sont importants pour les divers rapports entre calcium et élastine et servent, peut-être, comme centres de nucléation au cours de la calcification de la protéine.
    Abstract: Zusammenfassung Vorgängige Studien haben die Bedeutung der Carboxyl-, Sulfhydryl- und Aminogruppen als Stellen der Calciumbindung im Elastin gezeigt. Die vorliegende Arbeit hatte zum Ziel, die Rolle der neutralen Koordinationsstellen im Elastin als mögliche Calcium-Bindungsseite abzuklären. Die Calciumbindindung an das Elastin wurde durch solche Lösungsmittelveränderungen erhöht, die auch gleichartige Verschiebungen im Proteinmolekül bewirkten. In Methanol-Wasser-Mischungen schien die Calciumbindung nicht von Veränderungen des pH oder der Ionenstärke abhängig zu sein. Von 10 Ionen, bei welchen die Bindung überprüft wurde, war einzig diejenige des Calciums signifikant erhöht, wenn Methanol zugesetzt wurde. Es wird vorgeschlagen, daß die neutralen Stellen für die verschiedenen Vorgänge, bei welchen Calcium und Elastin beteiligt sind, eine wichtige Rolle spielen und vielleicht für die Verkalkung der Proteine als Nukleationszentren in Frage kommen.
    Notes: Abstract Previous studies have implicated carboxyl groups, sulphhydryl groups and amino groups as the sites for calcium binding in elastin. In this study, the concept was investigated that neutral co-ordinating sites in elastin may also provide calcium binding sites. Calcium binding to elastin was increased upon solvent changes which also effected conformational changes in the protein. In methanol-H2O mixtures calcium binding appeared to be independent of changes in pH and ionic strength. Of ten ions tested (Ca2+, Co2+, Na+, Cu2+, Zn2+, Cr3+, Pb+2, K+, Rb+, and Mg2+), only calcium binding was significantly increased when methanol was added. It is proposed that neutral sites are important to the various relationships involving calcium and elastin and perhaps serve as nucleation centers in the calcification of the protein.
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    Calcified tissue international 15 (1974), S. 31-44 
    ISSN: 1432-0827
    Keywords: Calcification ; Regeneration ; Repair ; Shell ; Mollusc
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Une régénération de la coquille induite expérimentalement chez l'excargot terrestreOtala lactea a été utilisée pour l'étude du substratum sur le type de régénération minérale. Des subsrats, comportant le periostracum de quatre espèces de mollusques, des répliques de surfaces et la membrane externe de la coquille de poule, ont été insérés au niveau de la région de reconstitution de la coquilled'Otala. Les divers types de dépôt de carbonate de calcium sont étudiés en microscopie électronique par balayage. Les types minéraux, formés sur les substrats mis en place, se conforment à la microtopographie des surfaces et se trouvent être nettement différents du type minéral observé au cours d'une régénération normale de la coquille. Les types minéraux de la surface minérale externe normale de la coquille de quatre espèces de mollusques semblent identiques aux types individuels de surface du periostracum sur lequel les cristaux de la coquille se déposent. Il semble que ce type de surface minérale externe de la coquille se forme par l'interaction de cristaux de CaCO3 qui se développent dans une matrice organique au contact du periostracum. La coquille normaled'Otala est constituée d'aragonite à orientation hautement préférentielle. La coquille régénérée contient de la calcite et de l'aragonite orientées au hasard. Le rapport aragonite-calcite est très variable et parait indépendant des substrats expérimentaux placés dans la région de régénération.
    Abstract: Zusammenfassung Der Einfluß der Unterlage auf die Art der Mineral-Neubildung im Gehäuse der Landschnecke,Otala lactea, wurde mit dem Raster-Elektronenmikroskop untersucht, indem in der Regenerationszone Fremdkörper eingeschoben wurden. Diese Fremdkörper waren Periostraca von vier anderen Molluskenarten, Plastikabdrücke von Periostraca oder der äußeren Mineral-Oberfläche normaler Gehäuse und die äußere Membran einer Hühnereierschale. Die Oberflächenstruktur des Kalziumkarbonats, das auf diesen Unterlagen abgelagert wurde, entsprach der Mikrotopographie der Unterlags-Oberfläche und war deutlich verschieden von der Mineraloberflächenstruktur bei normaler Regenerierung des Gehäuses. Die Strukturen der normalen Mineraloberflächen der vier Molluskenarten entsprachen der Struktur der Periostracum-Oberfläche, auf welcher die Kalziumkarbonat-Kristalle abgelagert werden. Die Mineraloberflächenstruktur des Gehäuses ergibt sich vermutlich durch die Wechselwirkung zwischen den Kalziumkarbonat-Kristallen, welche in einer organischen Matrix wachsen und der Berührung mit dem Periostracum. Das normale Mineral von Otala-Gehäusen besteht aus Aragonit in ganz bestimmter Anordnung. Regeneriertes Gehäuse enthielt Kalzit in zufälliger Anordnung und Aragonit, meist auch in zufälliger Anordnung. Das Verhältnis Aragonit/Kalzit variierte stark und schien unabhängig von der experimentellen Unterlage zu sein.
    Notes: Abstract Experimentally induced shell repair in the land snailOtala lactea has been employed to study the influence of the substratum on microtopography of the surface of mineral deposited by molluscs. Substrata, including the periostraca of four species of molluscs, surface replicas, and the outer membrane of the eggshell of the hen, have been inserted in the area of shell repair inOtala, and the topographical patterns of deposition of calcium carbonate have been observed by scanning electron microscopy. The mineral patterns formed on all inserted substrata conformed to the microtopography of the surfaces and were distinctly different from that of normal shell regeneration. The topography of the normal outer mineral surface of the shell of four species of molluscs were observed to conform to the topography of the periostraca on which the crystalline shell is deposited. It is inferred that the topography of the outer mineral surface of the shell probably results from the interaction of crystals of aragonite and calcite growing in an organic matrix and in contact with the periostracum. Normal shell ofOtala consists of fine-grained aragonite in highly preferred orientation. Regenerated shell contains calcite grains in random orientation and aragonite, usually in random orientation. The aragonite-to-calcite ratio varies widely and appears to be independent of experimental substrata placed in the area of repair.
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    Calcified tissue international 14 (1974), S. 229-244 
    ISSN: 1432-0827
    Keywords: Teeth-enamel ; Histocytochemistry ; Calcification
    Source: Springer Online Journal Archives 1860-2000
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    Description / Table of Contents: Résumé Des études histochimiques et d'affinités tinctoriales sont menées sur des germes dentaires de foetus de veauxin situ, fixés dans du chlorure de cyanure ou du formol. La concentration en protéine décroit avec la maturation de l'émail. Le tissue est divisé en deux bandes distinctes: une région externe d'émail nouvellement formé et une région interne d'émail en maturation. L'émail mature, défini antérieurement par méthode biochimique, n'a pas été étudié. L'histidine, présente dans l'émail, est aisément traitée par l'iode: le traitement au benzol n'a pas d'effet. La tyrosine est mise en évidence par deux méthodes dans les deux types d'émail et décroit légèrement au cours de la maturation. La transition est progressive. Des groupements sulfhydriles sont présents dans l'émail jeune, mais non dans l'émail en voie de maturation, avec une transition nette entre les deux zones. Le tryptophane est présent dans toute l'épaisseur de l'émail. L'arginine ne réagit pas bien par la méthode de Sakaguchi, mais on peut la mettre en évidence en bloquant par la fluorodinitrobenzène les colorants anioniques. La lysine est présente et diminue avec la maturation. Les résidus d'acide carboxyliques sont mis en évidence dans les deux types d'émail, mais des bandes distinctes et moins de résidus sont observés au cours de la maturation. Les affinités tinctoriales anioniques à diverses concentrations de sels montrent que la concentration critique en électrolytes de l'émail est relativement élevée et semblable à celle de la kératine. Les affinités tinctoriales cationiques indiquent que l'ionisation des groupements d'acide carboxylique peut être supprimée dans l'émail jeune, mais non dans l'email en voie de maturation. L'émail jeune a les caractéristiques histochimiques d'une kératine, mais non pas l'émail mature, qui ressemble à un groupe de protéines globulaires ectodermiques.
    Abstract: Zusammenfassung Histochemische Methoden und Farbbindungs-Untersuchungen wurden an Zahnkeimen von Kalbsembryonen in situ, welche in Cyanurchlorid oder Formalin fixiert worden waren, ausgeführt. Die Proteinkonzentration nahm mit dem Reiferwerden des Schmelzes ab. Das Gewebe wurde in zwei genau abgegrenzte Bänder unterteilt, einen äußeren Bereich aus neu gebildetem Schmelz und einen inneren Bereich aus reifendem Schmelz. Was früher biochemisch als reifer Schmelz definiert wurde, wurde nicht untersucht. Histidin lag im Schmelz vor und konnte leicht iodiert werden, eine Benzoilierung fand jedoch nicht statt. Zwei verschiedene Tyrosinanalysen bestätigen dessen Gegenwart in neugebildetem und reifendem Schmelz, wobei es aber beim Reifeprozess etwas abnahm. Der Übergang zeigte sich fließend. Sulphydryl-Gruppen waren in neugebildetem, aber nicht in reifendem Schmelz vorhanden, wobei sich ein abrupter Übergang zwischen den beiden Zonen fand. Tryptophan konnte durch die gesamte Schmelzbreite nachgeweisen werden. Argininrückstände reagierten kaum auf die Sakaguchi-Methode, aber sie konnten durch die anionische Farbstoffe blockierende Fluorodinitrozenzen-Methode nachgewiesen werden. Lysin war anwesend und nahm im Laufe des Reifeprozesses ab. Carboxylsäure-Rückstände wurden in reifendem und neugebildetem Schmelz nachgewiesen, jedoch mit scharfer Abgrenzung und deutlich weniger Rückständen in reifendem Schmelz. Die Bindung mit anionischen Farbstoffen bei verschiedenen Salzkonzentrationen zeigte, daß die kritische Elektrolyten-Konzentration nicht besonders hoch und derjenigen von Keratin ähnlich ist. Untersuchungen mit kationischen Farbstoffen deuteten an, daß die Ionisierung von Carboxylsäure-Gruppen in neugebildetem, nicht aber in reifendem Schmelz, unterdrückt werden kann. Neugebildeter Schmelz hat die histochemische Eigenschaften von Keratin; reifender Schmelz hingegen hat diese Eigenschaft nicht und gleicht eher einem Vertreter der Gruppe der ektodermalen globulären Proteine.
    Notes: Abstract Histochemical methods and dye binding studies were carried out on foetal calf tooth germsin situ fixed in cyanuric chloride or formalin. Protein concentration decreased upon maturation of enamel. The tissue was divided into two distinct bands, an outer region of newly formed enamel and an inner region of maturing enamel. What has previously been defined biochemically as mature enamel was not investigated. Histidine was present in enamel and easily iodinated but protected from benzoylation. Two methods for tyrosine analysis confirm its presence in both newly formed and maturing enamel but with some decrease upon maturation. The transition was gradual; Sulphydryl groups were present in newly formed, but not in maturing, enamel, with abrupt transition between the two zones. Tryptophane was demonstrated throughout the entire thickness of enamel. Arginine residues were not very reactive to Sakaguchi's method but they were demonstrable by fluorodinitrobenzene blockade of anionic dyes. Lysine was present, decreasing upon maturation. Carboxylic acid residues were demonstrated in both maturing and newly formed enamel but with distinct banding and noticeably fewer residues in maturing enamel. Anionic dye binding at various salt concentrations showed that the critical electrolyte concentration of enamel is moderately high and similar to that of keratin. Cationic dye binding studies indicated that ionization of carboxylic acid groups in newly formed enamel, but not maturing enamel, might be suppressed. Newly formed enamel has the histochemical characteristics of a keratin but maturing enamel does not, more nearly resembling one of a group of ectodermal globular proteins.
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    Calcified tissue international 16 (1974), S. 93-107 
    ISSN: 1432-0827
    Keywords: Dentinogenesis ; Globules ; Pyrophosphatase ; Calcification ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Three-day-old rats were fixed by perfusion with glutaraldehyde and thin slices were cut of the first molar germs. The slices were treated with EDTA and “activated” with buffered solutions containing Mg2+, Ca2+ or Zn2+. Incubation was carried out in buffered solutions (pH 8.5) containing inorganic pyrophosphate and Pb2+. In the Mg2+-activated specimens incubation products were localized to the plasma membranes in the stratum intermedium and the subodontoblastic area. Lead deposits were found on the periphery of the dentinal globules. Incubation products were more randomly distributed in Ca2+-activated specimens whereas those activated with Zn2+ displayed a deposition of lead precipitates mainly corresponding to that seen after activation with Mg2+. The findings are discussed in reference to the localization of alkaline phosphatase in the dentin-producing tissues and it is proposed that the results are indicative of the presence of an inorganic pyrophosphatase in these tissues.
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    Calcified tissue international 16 (1974), S. 169-182 
    ISSN: 1432-0827
    Keywords: Histochemistry ; Alkaline phosphatases ; Calcification ; Bone ; Teeth
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Activity of alkaline phosphatases in unfixed cold microtome setions from the lower first molar area of newborn mice was recorded by histochemical methods. A substrate specificity test included the following phosphate compounds: ATP, CTP, GTP, UTP, ADP, AMP, GP, PPi, MDP and naphthol AS-TR phosphate. Intense staining was obtained in osteoblasts, stratum intermedium of the enamel organ and odontoblasts with all the substrates, except PPi and MDP. Staining of skeletal muscle fibres was obtained only with triphosphates as substrates. Addition of-SH groups decreased the hydrolysis of triphosphate compounds in cells involved in mineralization while the hydrolysis of monophosphate was inhibited. In contrast triphosphatase activity in striated muscle was enhanced when-SH compounds were added. Demineralization with EDTA diminished the cytoplasmic staining but induced a nuclear staining in hard tissue forming cells when triphosphates were used as substrates. No cytoplasmic and only slight nuclear staining was seen with GP or AMP as substrates. The triphosphate hydrolyzing capacity of tongue muscle fibres was, however, increased after the decalcification treatment. Addition of Mg2+ ions to the incubation media distinctly lowered the hydrolysis of triphosphates in the investigated tissues whereas the hydrolysis of ADP, AMP, GP and naphthol AS-TR phosphate remained unchanged. In view of the findings the triphosphatase activities at alkaline pH of muscle fibres and of cells related to hard tissue formation are considered to be due to activity of separate enzymes. The orthophosphate liberating enzyme activities at alkaline pH in osteoblasts, stratum intermedium and odontoblasts may be expressions of the catalytic functions of one common enzyme. Furthermore, the results indicate that CaATP might be the substrate used by the alkaline ATPase in mineralizing areas.
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    Keywords: Dentin ; Calcification ; Pyrophosphate ; Enzyme
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    Description / Table of Contents: Résumé Après incubation de coupes de germes de molaires de rats, fixés aux aldéhydes, dans une solution alcaline de pyrophosphate de plomb, une réaction positive de coloration au sulfure est localisée au niveau du front de minéralisation de la dentine, dans le stratum intermedium et les cellules sous-odontoblastiques. Au microscope électronique, le nombre et la taille de cristaux, en forme d'aiguilles, sont augmentés dans la zone en voie de minéralisation (denture intermédiaire). Un fin précipité est surtout observé dans les noyaux du stratum intermedium et les cellules sous-odontoblastiques. Les effects de l'EDTA, Zn2+, Ca2+, Mg2+ et citrate sont aussi étudiés et la probabilité d'activité en pyrophosphatase inorganique est discutée en fonction des dépôts de plomb observés. L'action d'une pyrophosphatase inorganique sur la précipitation du phosphate de plomb ne peut être exclue. Cependant, la réaction de coloration peut être aussi due à une absorption non catalysée de composé de plomb sur les cristaux d'apatite.
    Abstract: Zusammenfassung Schnitte von Ratten-Molarkeimen, die mit Aldehyd fixiert und in einer alkalischen Blei-Pyrophosphat-Lösung inkubiert wurden, zeigten an folgenden Stellen eine positive Sulfid-Färbungsreaktion: in der Mineralisierungsfront des Dentins, im Stratum intermedium, und in den subodontoblastischen Zellen. Elektronenmikroskopisch waren Anzahl und Größe der sichtbaren, nadelartigen Kristallite in der Mineralisationszone (intermediäres Dentin) erhöht. Ein feiner Niederschlag konnte vorwiegend in den Nuclei des Stratum intermedium und der subodontoblastischen Zellen beobachtet werden. Es wurde zudem die Wirkungsweise von EDTA, Zn2+, Ca2+, Mg2+ und Zitrat untersucht. Schließlich wird die Wahrscheinlichkeit einer Aktivität von anorganischer Pyrophosphatase im Zusammenhang mit den beobachteten Bleiablagerungen besprochen. Daß anorganische Pyrophosphatase bei der Fällung von Bleiphosphat eine Rolle Spielt, ist nicht auszuschließen. Allerdings kann die Farbreaktion auch durch nicht-katalysierte Adsorption der Bleiverbinding an die Apatitkristalle bedingt sein.
    Notes: Abstract After incubation of aldehyde-fixed sections of rat molar germs in an alkaline solution of lead pyrophosphate, a positive sulphide staining reaction was observed in the mineralization front of the dentin, in the stratum intermedium, and in the subodontoblastic cells. In the electron microscope, the number and size of visible needle-like crystallites was increased in the mineralization zone (intermediate dentin). A fine precipitate was observed predominantly in the nuclei of the stratum intermedium and subodontoblastic cells. The effects of EDTA, Zn2+, Ca2+, Mg2+ and citrate were also studied and the probability of inorganic pyrophosphatase activities is discussed in relation to the observed lead deposits. The interaction of an inorganic pyrophosphatase cannot be excluded in the precipitation of lead phosphate. However, the staining reaction may also be due to uncatalyzed adsorption of the lead compound onto the apatite crystals.
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    Calcified tissue international 15 (1974), S. 201-212 
    ISSN: 1432-0827
    Keywords: Ultrastructure ; Alkaline Phosphatase ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Des cals expérimentaux de neuf jours, formés au niveau de radius de jeunes rats, sont traités par la méthode calcium-cobalt de Gomori (1939) pour la mise en évidence ultrastructurale de la phosphatase alcaline afin d'étudier son rôle éventuel dans le dépôt du calcium. L'activité enzymatique apparait initialement sous forme de précipités globulaires en dehors de la membrane cellulaire de jeunes chondroblastes hypertrophiques. Ce précipité donne ensuite naissance à des corps sphériques de phosphatase alcaline qui se forme près de la cellule. Ces corps sphériques s'observent dans une zone intermédiaire plus éloignée. Une formation de cristaux en aiguilles (apparemment une calcification) se développe dans des corps isolés ou agrégés, laissant voir nettement leurs limites, même lorsque la calcification est plus avancée au point qu'on ne peut plus distinguer des cristaux individuels. Au niveau des coupes témoins, traitées de façon identique mais sans substrat ou avec de l'E.D.T.A., on n'observe ni précipité enzymatique ou corps sphériques. L'aspect des dépôts cristallins dans des corps qui contiennent de la phosphatase alcaline ne peut s'expliquer que par l'existence d'une association étroite entre enzymes et calcification.
    Abstract: Zusammenfassung Neun Tage alter experimenteller Kallus an Radii von jungen Ratten wurde mit Gomori's (1939) Calcium-Kobalt Methode untersucht, um die Verteilung der alkalischen Phosphatase und ihre Beziehung zur Calciumablagerung ultrastrukturell zu demonstrieren. Enzymaktivität zeigte sich zuerst als globulares Präzipitat außerhalb der Zellmembran von Knorpelzellen im Beginn der Hypertrophie. Aus dieser Präzipitatschicht entstanden dann gerundete Körperchen, die sich von der Zelle abtrennten. Solche Körperchen wurden auch in größerer Entfernung von der Zelle beobachtet, d.h. in einer Zwischenzone zwischen benachbarten Zellen. Nadelförmige Kristalle, wahrscheinlich von Calcium-Salzen, wurden in einzelnen oder aggregierten Körperchen beobachtet. Die äußere Zone der Körperchen blieb jedoch deutlich sichtbar, selbst dann, wenn der Calciumgehalt derart zugenommen hatte, daß einzelne Kristalle nicht länger erkennbar waren. In Kontrollen, die in gleicher Weise behandelt waren, aber ohne Substrat oder mit Zufügung von EDTA, wurden weder Präzipitate noch Körperchen beobachtet. Das Auftreten von Calciumablagerungen in alkalischer Phosphatase enthaltenden Körperchen scheint kaum anders erklärbar als durch eine enge funktionelle Verbindung zwischen Enzym und Calciumablagerung.
    Notes: Abstract Nine day old experimental calluses in radii of young rats were treated with Gomori's (1939) calcium-cobalt method to demonstrate ultrastructurally the presence of alkaline phosphatase in a search for its possible role in the desposition of calcium. Enzyme activity first appeared as globule-like precipitates outside the cell membrane of early hypertrophic cartilage cells. This precipitate layer then seemed to give rise to spherical bodies of alkaline phosphatase which occur at a slight distance from the cell. The spherical bodies were also observed further away from the cell in an intermediate zone between neighboring cells. Needle-like crystal formation, apparently calcification, occurred inside single or aggregated bodies, leaving their peripheral rim clearly visible, even when calcification had increased to such an extent that individual crystals could no longer be recognised. In controls, treated in the same way but without substrate, or with EDTA, no enzyme precipitate or spherical bodies were seen. The appearance of crystalline deposits in bodies which contain alkaline phosphatase seems difficult to explain on any other basis than that there is a close functional association between the enzyme and calcification.
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    Calcified tissue international 16 (1974), S. 59-71 
    ISSN: 1432-0827
    Keywords: Apatite ; Calcification ; Pineal gland ; Calcospherulites ; Calculi
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Calcifications of the pineal gland in the form of calcospherulites have been studied by optical microscopy, electron microscopy, electron probe analysis, X-ray diffraction, thermogravimetry, and infra-red and chemical analysis. Complex calcospherulite textures have been observed which have a granular substructure made up of apatite crystals averaging 218 Å in length and 38 Å in width. These apatite crystals appear to be a carbonate-containing hydroxyapatite, mineralogically similar to enamel.
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    Calcified tissue international 16 (1974), S. 109-127 
    ISSN: 1432-0827
    Keywords: Dentinogenesis ; Diphosphonates ; Calcification ; Collagen ; Electron Microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract The effects of high doses of ethylene-1-hydroxy-1,1-diphosphonate (EHDP) were investigated at the light microscopic and subcellular level. The administration of EHDP at a concentration of 7.5–10 mg P/kg body weight/day over a short period of time resulted in complete inhibition of crystal formation in predentin and pre-enamel. An increased predentin width was observed and within newly-formed predentin areas the formation ofcollagen fibrils was grossly disturbed. In addition, fine precipitates appeared in the ground substance. The presence of unusual thread-like elements within specific bodies in the cytoplasm of the odontoblastic processes may be indicative of an interference by EHDP in e.g. the synthesis of precollagen. The possibility of an inhibition by EHDP of the extracellular aggregation of collagen molecules is also discussed. EHDP further inhibited crystal formation within dentinal globules. Functioning ameloblasts were grossly affected in EHDP-treated rats, and it is suggested that this is related to an inhibition of crystal formation in pre-enamel.
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    Calcified tissue international 13 (1973), S. 173-185 
    ISSN: 1432-0827
    Keywords: Calcification ; Apatite ; Whitlockite ; Amorphous
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Des études cristallographiques, spectroscopiques et chimiques ont permis de mettre en évidence deux types distincts de phosphate de calcium dans les tissus de patients, atteints de troubles rénaux chroniques. Les dépôts des calcifications périarticulaires sont des apatites contenant des carbonates, alors que ceux trouvés dans les tissus viscéraux calcifiés sont soit des micro-cristaux de whitlockite de magnésium ou un précurseur de ce composé. Les clichés de diffraction aux rayons X sont de type apatitique pour les calcifications tumorales et amorphe pour les calcifications viscérales. Il semble que la présence de magnésium facilite la formation de ce dernier type en inhibant la cristallisation de l'apatite et en stabilisant la phase amorphe comme dans les systèmes synthétiques.
    Abstract: Zusammenfassung Kristallographische, spektroskopische und chemische Untersuchungen belegen die Anwesenheit von zwei bestimmten Arten von Calciumphosphat-Ablagerungen in den Geweben von Patienten mit chronischem Nierenversagen. Die Ablagerungen, welche in periartikulären Verkalkungen gefunden wurden, sind Karbonat-enthaltendes Apatit; diejenigen in visceralen Geweben sind entweder Kleinkriställchen von Magnesium-Whitlockit oder eine direkte Vorstufe davon. Röntgendiffraktionsspektren dieser beiden Ablagerungsarten zeigen, daß tumorale Verkalkungen apatitartig und viscerale Verkalkungen amorph sind. Es wird vorgeschlagen, daß die Anwesenheit von Magnesium die Bildung dieser zweiten Art der Calciumablagerung begünstigt, indem es die Apatitbildung stört und die amorphe Phase stabilisiert, was in synthetischen Systemen beobachtet werden konnte.
    Notes: Abstract Crystallographic, spectroscopic and chemical studies demonstrate the presence of two distinct types of calcium phosphates deposits in tissues of patients with chronic renal failure. Deposits found in periarticular calcifications are carbonate-containing apatite, while those found in calcified visceral tissues are either microcrystallites of magnesium whitlockite or an immediate precursor of this compound. X-ray diffraction patterns of these two types show tumoral calcification to give an apatitic pattern and visceral calcification, an amorphous one. It is suggested that the presence of magnesium promotes the formation, of this latter type of calcium deposit by disturbing the crystallization of the apatite and stabilizing the amorphous phase as observed in synthetic systems.
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    Calcified tissue international 12 (1973), S. 117-124 
    ISSN: 1432-0827
    Keywords: Mouse ; Calcification ; Metals ; Mast Cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Des souris, ayant reçu par voie intra-veineuse une solution d'acétate de plomb, sont injectés par voie sous-cutanée avec du sulfate de polymixine B. Au niveau de ce dernier point d'injection, on observe des calcifications. L'examen histologique montre une dégranulation des mastocytes, une vaso-dilatation locale ainsi qu'une diffusion d'ions à partir des vaisseaux. Après 24 heures, des ions calcium et phosphate sont présents. En outre. des groupes de souris, pré-injectées par voie intraveineuse avec de l'acétate de plomb, reçoivent des extraits de granules de mastocytes, comportant du phosphate d'histamine, du sulfate de créatinine sérotonine, de l'héparine, du sulfate de chondroitine ou de l'acide hyaluronique. Seules les injections de sérotonine et d'histamine, connues comme vaso-dilatateurs et augmentant probablement la perméabilité capillaire, provoquent des calcifications. Il semble que le rôle des mastocytes dans la calcification soit lié principalement à la production d'une vasodilatation locale, avec perméabilité capillaire augmentée, plutot qu'à une action de liaison initiale des ions métalliques.
    Abstract: Zusammenfassung Nachdem Mäusen Bleiacetatlösungen intravenös eingespritzt worden waren, erhielten die Tiere Polymixin-B-Sulfat subkutan. Dies führte zu Verkalkung an der zweiten Injektionsstelle. Die histologische Untersuchung zeigte eine deutliche degranulation der Mastzellen, eine lokale Vasodilatation und das Austreten von Bleiionen aus diesen Gefäßen 24 Std nach der Injektion konnte eine groß Menge von Calcium- und Phosphationen eindeutig nachgewiesen werden. Weiter wurden Gruppen von Mäusen, die vorher Bleiacetat intravenös erhalten hatten. Bestandteile von Mastzellen-Granula injiziert. Diese bestanden aus Histaminphosphat, Serotonin-Creatinin-Sulfat, Heparin, Chondroitin-Sulfat oder Hyaluronsäure. Unter diesen exogenen Bestandteilen bewirkten nur Serotonin und Histamin eine Verkalkung. Beide sind starke Vasodilatatoren und erhöhen vermutlich die kapillare Permeabilität. Es wird postuliert, daß die Rolle der Mastzelle in der “calcergy” viel eher auf einer lokalen Vasodilatation mit erhöhter kapillarer Permeabilität als auf einer primären Bindung der Metallionen beruht.
    Notes: Abstract Mice previously injected intravenously with lead acetate solution were injected subcutaneously with Polymixin B Sulphate. This resulted in calcification at the latter injection site. Histological evidence showed marked mast cell degranulation, local vasodilation and the efflux of lead ions from these vessels. At twenty four hours after injection abundant calcium and phosphate ions were readily demonstrated. In addition, groups of mice pre-injected intravenously with lead acetate, were injected with constituents of mast cell granules. These included histamine phosphate, serotonin creatinine sulphate, heparin, chondroitin sulphate or hyaluronic acid. Of these exogenous constituents used, calcification was found only after serotonin and histamine injections, both of which are potent vasodilators and presumbly increase capillary permeability. It is postulated that the role of the mast cell in calcergy may be mainly that of producing local vasodilation with increased capillary permeability rather than the initial binding of metallic ions.
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    Calcified tissue international 12 (1973), S. 169-173 
    ISSN: 1432-0827
    Keywords: Calcification ; Histochemistry ; Autoradiography ; Fetal ; Mandible
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Des rats femelles reçoivent du45Ca ou du32P au 13ème et 18ème jour de la grossesse. Les coupes sériées des têtes de foetus sont étudiées par autoradiographie et par diverses méthodes histologiques pour déterminer la calcification. La détection la plus précoce de45Ca s'observe simultanément comme une réaction positive pour le calcium avec une des méthodes histologiques utilisées.32P est en évidence par les méthodes autoradiographiques un peu plus tard que le45Ca et sa présence coincide avec la réaction positive la plus précoce observée avec les autres méthodes histologiques utilisées. Les os de ces têtes foetales commencent à se calcifier selon un mode particulier commençant par la mandibule, puis l'os frontal et le maxillaire supérieur, suivis par les os nasaux, pariétaux et interpariétaux.
    Abstract: Zusammenfassung Weibliche Ratten erhielten45Ca und32P zwischen dem 13.–18. Tag der Schwangerschaft. Zur Ermittlung der Verkalkung wurden Serienschnitte der Köpfe ihrer Feten mittels Autoradiographie sowie durch verschiedene histologische Methoden untersucht. Das erste Auftreten von45Ca war ebenfalls von einer positiven Calciumreaktion mittels einer der verwendeten histologischen Methoden begleitet.32P wurde in der Autoradiographie erst etwas später als45Ca festgestellt und dessen Nachweis deckte sich mit den ersten positiven Reaktonen aller anderen verwendeten histologischen Methoden. Die Knochen in diesen Fetusköpfen begannen in einer bestimmten Sequenz zu verkalken: zuerst der Unterkiefer, dann das Stirnbein und der Oberkiefer, dann das Nasenbein, die Parietal- und Interparietalknochen.
    Notes: Abstract Female rats were given45Ca or32P from 13 to 18 days of pegnancy. Serial sections from the heads of their fetuses were studied by autoradiography, as well as by several histological methods for assessing calcification. The earliest detection of45calcium occured at the same time as a positive reaction for calcium with one of the histological methods used.32P was not detectable by autoradiographic methods until somewhat later than45Ca, and its presence coincided with the earliest positive reaction with all of the other histological methods employed. The bones in these fetal heads began to calcify in a partcular sequence, the mandible first, then the frontal bone and maxilla, followed by the nasal, parietal and interparietal bones.
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    Calcified tissue international 13 (1973), S. 63-72 
    ISSN: 1432-0827
    Keywords: Structure ; Poly(glycol monomethacrylate) ; Gel ; Implants ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Abstract L'effet de la structure physique et des modification chimiques de gels de poly(glycol monométhacrylate) sur la calcification d'implants tests a été étudié. Alors que les modifications chimiques du poly(glycol monométhacrylate) osseux, obtenu par introduction de groupes formateurs d'ions, n'affectent pas le processus de calcification, on observe un effet net de la structure physique du polymère par action du rapport monomère: eau sur le processus de calcification des implants. Des gels homogènes et microporeux montrent rarement une calcification autor de l'implant; dans les gels macroporeux, la calcification s'observe massivement autour de l'implant, en cas de porosité élevée en son centre. Ce phénomène a été expliqué par la pénétration diverse par du tissu néoformé ainsi que par la possibilité de pénétration d'éléments nutritifs cellulaires, liée à la porosité de l'implant.
    Abstract: Zusammenfassung Die Wirkung der physikalischen Struktur und der chemischen Veränderung von Poly-(Glycol-Monomethacrylat)-Gelen auf die Verkalkung von Implantat-Proben wurde untersucht. Die chemische Veränderung der Poly(Glycol-Monomethacrylat)-Grundstruktur, hervorgerufen durch das Beifügen ionogener Gruppen, hatte auf den Verkalkungsprozeß keine Wirkung; die physikalische Struktur des Polymers hingegen hatte eine markante Wirkung auf den Verkalkungsprozeß der Implantate, welche durch das Verhältnis Monomer: Wasser bestimmt wurde. Homogene und kleinporöse Gele zeigten ganz ausnahmsweise eine Verkalkung am Rande des Implantats; in großporösen Gelen erfolgte massive Verkalkung am Rande des Implantats, bei höherer Porosität in dessen Zentrum. Dieses Phänomen wird zurückgeführt auf den unterschiedlichen Grad, in welchem das neu gebildete Gewebe das Implantat durchdringt und auf die verschiedene Zugänglichkeit der Nahrung der eindringenden Zellen, welche von der Porosität des Implantats abhängt.
    Notes: Abstract The effect of the physical structure and chemical modification of poly(glycol monomethacrylate) gels on the calcification of test implants has been studied. While the chemical modification of the poly(glycol monomethacrylate) backbone by the introduction of ionogenic groups did not affect the process of calcification, there was a substantial effect of the physical structure of the polymer, as determined by the monomer: water ratio, on the process of calcification of the implants. Homogenous and microporous gels showed calcification in the margin of the implant only exceptionally; in macroporous gels calcification occurred massively in the margin of the implant and in the case of a higher porosity in its centre. This phenomenon has been explained by the different degree to which the implant is penetrated by the newly formed tissue, as well as by the difference in accessibility of nutrition to the penetrating cells depending on the porosity of the implant.
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    Calcified tissue international 12 (1973), S. 209-216 
    ISSN: 1432-0827
    Keywords: Analysis ; Osteon ; Bone ; Composition ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Une étude microscopique de la composition du tissu osseux, réalisée initialement au niveau des ostéones, a été étendue à l'os lamellaire interstitiel périosté et aux ≪Mittellinien≫. Le phosphore, calcium, l'azote total, les mucopolysaccharides (sous forme d'hexosamines) et le collagène (sous forme d'hydroxyle-proline) ont été déterminés dans ce tissu au point de vue pondéral et volumétrique. Le calcium a également été déterminé dans les ostéones aux stades initial et final de la calcification. Malgré des rapports Ca/P sensiblement identiques, l'os lamellaire interstitiel périosté et les Mittellinien diffèrent en composition organique. L'os interstitiel a un contenu plus élevé en azote (absolu et par rapport au phosphore) et en hydroxyle-proline et azote non-collagénique que les Mittellinien. En étudiant les résultats sur la composition du tissu ostéoide, les ostéones à divers degrés de calcification et les structures décrites ci-dessus, on se rend compte de la composition variable du tissu osseux d'une plage à l'autre. D'après les résultats analytiques, il semble que dans tout processus de calcification rapide, des protéines non-collagéniques (probablement des protéoglycanes) sont présentes.
    Abstract: Zusammenfassung Eine Untersuchung der Zusammensetzung des Knochengewebes auf der Stufe mikroskopischer Strukturen, welche früher auf die Haversianischen Systeme begrenzt war, ist auf den interstitiellen Lamellenknochen des Periosts und auf die Mittellinien ausgedehnt worden. Phosphor, Calcium, Gesamtstickstoff, Mucopolysaccharide (als Hexosamine) und Kollagen (als Hydroxyprolin) wurden in diesen Materialien in bezug auf Gewicht und Volumen bestimmt. Calcium wurde auch in den Osteonen in der Anfangs- und Endphase der Verkalkung bestimmt. Trotz praktisch identischem Ca/P-Verhältnis unterscheiden sich interstitieller Lamellenknochen des Periosts und Mittellinien in ihrer organischen Zusammensetzung. Interstitieller Knochen hat einen höheren Gesamtstickstoffgehalt (absolut sowie auf Phosphorgehalt bezogen), ebenso mehr Hydroxyprolin und nicht-kollagenen Stockstoff als die Mittellinien. Eine Zusammenstellung von Daten über die Zusammensetzung des Osteoid-Gewebes, der Osteonen in verschiedenen Stadien der Verkalkung und der oben aufgeführten Strukturen zeigt, wie unterschiedlich die Zusammensetzung des Knochengewebes sogar von einer mikroskopischen Struktur zur angrenzenden sein kann. Es wurde versucht, aus den analytischen Daten eine allgemeine Regel betreffend den verkalkungsprozess zu erhalten, nämlich: Wo immer eine Verkalkung rasch stattfindet, sind nicht-kollagene Proteine (möglicherweise Proteoglycane) anwesend.
    Notes: Abstract A study of the composition of bone tissue at the level of microscopic structures, previously limited to the Haversian systems, has been extended to interstitial periosteal lamellar bone and to the “Mittellinien”. Phosphorus, calcium, total nitrogen, mucopolysaccharides (as hexosamines) and collagen (as hydroxyproline) were determined in these materials on both weight and volume basis. Calcium was also been determined in osteones at the initial and final stage of calcification. In spite of virtually identical Ca/P ratios, interstitial periosteal lamellar bone and Mittellinien differ in their organic composition. Interstitial bone has a higher total nitrogen content (both absolute and relative to phosphorus) as well as higher hydroxyproline and non-collagenous nitrogen than the Mittellinien. A compilation of data on the composition of osteoid tissue, osteones at different degrees of calcification and the above structures, shows how variable is the composition of bone tissue even from one microscopic structure to the one adjacent. Tentative indications of a general rule governing the process of calcification were obtained from the analytical data; namely that wherever calcification is taking place rapidly, non-collagenous proteins (possibly proteoglycans) are present.
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    Calcified tissue international 13 (1973), S. 335-337 
    ISSN: 1432-0827
    Keywords: Bone ; Morphogenesis ; Calcification
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    Calcified tissue international 10 (1972), S. 128-135 
    ISSN: 1432-0827
    Keywords: Bone ; Embryo ; Calcification ; Phosphate ; Pyrophosphate ; Carbonate ; Apatite
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé La maturation chimique et physique du sel osseux est étudiée par l'obervation des caractéristiques stoichiométriques et aux infra-rouges d'os embryonnaire d'oiseaux, du début jusqu'à la fin de la croissance adulte après éclosion. La succession des transformations chimiques de l'os en voie de développement montre surtout un rapport inverse entre le phosphate acide et le carbonate, ainsi que la formation de CO3-apatite. Ces résultats semblent indiquer que le CO 3 2− est substitué au HPO 4 2− au cours de la synthèse du CO3-apatite de l'os.
    Abstract: Zusammenfassung Die chemischen und physikalischen Umwandlungen zum stabilen Knochensalz in Hühnerknochen wurden anhand serienmäßiger Beobachtungen seiner stöchiometrischen Zusammensetzung und seiner infraroten Charakteristika untersucht; diese Beobachtungen erstreckten sich über die Zeit der frühen embryonalen Mineralablagerung bis zur vollständigen Reife nach dem Ausbrüten. Die Sequenz der chemischen Umwandlungen im sich entwickelnden Knochen zeigte hauptsächlich ein entgegengesetztes Verhältnis zwischen saurem Phosphat und Carbonat, das mit der Bildung von Carbonatapatit zusammenfällt. Diese Resultate weisen darauf hin, daß HPO 4 2− bei der Synthese von Carbonatapatit im Knochen durch CO 3 2− substituiert wird.
    Notes: Abstract The chemical and physical maturation of the bone salt was studied by serial observations on its stoichiometric and infrared characteristics in avian bone from early embryonic mineral deposition to full maturity after hatching. The sequence of chemical transformations in the developing bone showed most predominantly an inverse relationship between acid phosphate and carbonate, coincident with the formation of CO3-apatite. The data are consistent with the view that CO 3 2− is substituted for HPO 4 2− in the synthesis of CO3-apatite in bone.
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    Calcified tissue international 9 (1972), S. 67-79 
    ISSN: 1432-0827
    Keywords: Bone ; Calcium ; Calcification ; Mitochondria ; Cell
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Quatre vingt quinze pour cent du total du calcium et phosphate d'homogénats cellulaires, isolés mécaniquement de l'os, sédimentent avec des particules sub-cellulaires par centrifugation à 15000×G. Une méthode, destinée à isoler et à purifier partiellement cette fraction, riche en calcium, est mise au point par digestion enzymatique avec de la désoxyribonucléase et ultracentrifugation dans 70% de saccharose. Le fragment obtenu contient 10% de produit minéral, sous forme de complexe calcium-phosphate, et des protéines et lipides. Des quantités importantes de phospholipides et RNA sont notées, ainsi que des quantités faibles d'hexosamine et d'acide sialique. Les essais enzymatiques et la microscopie électronique indiquent que la fraction contient plusieurs types différents de particules cytophoniques, mais leur contenu en calcium n'a pu être déterminé.
    Abstract: Zusammenfassung Es wurde festgestellt, daß 95% des gesamten Calcium-und Phosphatgehaltes in Zellhomogenaten, welche mechanisch aus Knochen isoliert wurden, mit subzellulären Teilchen in einem Zentrifugenfeld von 15000×G sedimentieren. Zur Isolation und teilweisen Reinigung dieser calciumreichen Fraktion wurde eine Methode entwickelt, die auf enzymatischer Verdauung mit Desoxyribonuclease und Ultrazentrifugation mittels 70% Sucrose basiert. Der entstandene Niederschlag enthielt 10% Mineral, der in Form eines Calciumphosphat-Komplexes vorlag und einen hohen Gehalt an Protein und Lipid aufwies. Es fanden sich ferner signifikante Mengen von Phospholipid und RNS, dagegen nur kleine Mengen von Hexosamin und Sialsäure. Enzymbestimmungen und Elektronenmikroskopie zeigten, daß die gefundene Fraktion mehrere, sich voneinander unterscheidende Typen von Cytoplasmapartikeln enthielt; ihr Calciumgehalt konnte jedoch anhand der verfügbaren Resultate nicht bestimmt werden.
    Notes: Abstract Ninety-five percent of the total calcium and phosphate in homogenates of cells isolated mechanically from bone was found to sediment with subcellular particles in a centrifugal field of 15000×G. A method was devised for the isolation and partial purification of this calcium-rich fraction by enzymatic digestion with deoxyribonuclease and ultracentrifugation through 70% sucrose. The resultant pellet was 10% mineral, present as some form of calcium-phosphate complex, and was rich in protein and lipid. Significant amounts of phospholipid and RNA were present also, but only small amounts of hexosamine and sialic acid. Enzyme assays and electron microscopy indicated that the fraction contained several different types of cytoplasmic particles but those which contained calcium could not be determined from available data.
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    Calcified tissue international 9 (1972), S. 122-130 
    ISSN: 1432-0827
    Keywords: Tetracycline ; Development ; Calcification ; Statolith ; Nematocysts ; Aurelia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé L'effet de la tétracycline HCl sur la synthèse de statolithes de sulfate de calcium chezAurelia a été étudié. La tétracycline inhibe la synthèse de statolithes et nématocystes à un stade précoce de strobilation. La tétracycline, cependant, n'est pas incorporée dans les statolithes ou nématocystes en formation. Comme la tétracycline ne se combine pas avec le calcium des statolithes de sulfate de calcium dihydraté d'Aurelia, l'explication des effets d'inhibition sur la différenciation de statolithes et nématocystes ne semble pas liée avec un facteur en rapport avec l'incorporation. Des étudesin vitro de quatre systèmes inorganiques de calcium et de tétracycline montrent que le sulfate de calcium dihydraté (gypse) n'incorpore pas la tétracycline: il en est de même de son équivalent isostructural, le phosphate de calcium hydrogéné dihydraté (brushite). Le carbonate de calcium et le phosphate de calcium (apatite) incorpore la tétracycline. L'explication des différences de comportement du calcium peut être liée à la structure cristalline des composés respectifs, et, en particulier, au fait que l'ion Ca est prêt ou non à réagir avec la tétracycline.
    Abstract: Zusammenfassung Es wird über die Wirkung von Tetracyclinchlorhydrat auf die Synthese von Calciumsulfat-Statolithen beiAurelia berichtet. Wird das Tetracyclin in einem Frühstadium der Strobilation verabreicht, so hemmt es die Synthese der Statolithen und der Nematocysten. Das Tetracyclin wird jedoch nicht in die sich bildenden Statolithen oder Nematocysten eingebaut. Da sich das Tetracyclin nicht mit dem Calcium der Calciumsulfatdihydrat-Statolithen derAurelia verbindet, so kann dessen Hemmwirkung auf die Statolithen und die sich differenzierenden Nematocysten offenbar nicht mit einem einbaubedingten Faktor erklärt werden. Untersuchunge, die in vitro mit vier verschiedenen anorganischen Calciumsalzen und Tetracyclin ausgeführt wurden, zeigten, daß weder Calciumsulfatdihydrat (Gips), noch dessen isotrukturelles Aequivalent Calciumhydrogenphosphatdihydrat (Bruschit) Tetracyclin einbauen. Dagegen inkorporieren Calciumcarbonat und Calciumphosphat (Apatit) das Tetracyclin. Die Erklärung für dieses unterschiedliche Verhalten der Calciumsalze findet sich in der Kristallstruktur der betreffenden Verbindungen, d.h. es hängt davon ab, ob das Calciumion ür die Reaktion mit Tetracyclin leicht verfügbar ist.
    Notes: Abstract The effect of tetracycline HCl on synthesis of calcium sulphate statoliths inAurelia is reported. Tetracycline inhibits synthesis of statoliths and nematocysts when administered at an early stage of strobilation. The tetracycline, however, is not incorporated into the developing statoliths or nematocysts. As the tetracycline does not combine with the calcium of the calcium sulfate dihydrate statoliths ofAurelia, an explanation for its inhibitory effects on statoliths and nematocyst differentiation apparently does not rest with an incorporation-related factor. In vitro studies of four inorganic calcium systems and tetracycline revealed that calcium sulfate dihydrate (gypsum) did not incorporate tetracycline nor did its isostructural equivalent, calcium hydrogen phosphate dihydrate (brushite). Calcium carbonate and calcium phosphate (apatite) did incorporate tetracycline. The explanation for these different behaviors of calcium can be found in the crystal structure of the respective compounds, namely, whether or not the Ca ion is readily available to react with tetracycline.
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    Calcified tissue international 9 (1972), S. 109-121 
    ISSN: 1432-0827
    Keywords: Arthritis ; Phosphonates ; Bone ; Resorption ; Pathology ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé De l'adjuvant de Freund, constitué deMycobacterium butyricum tués, en suspension dans de l'huile minérale, provoque une arthrite chez les rats qui rappelle certaines formes d'arthrites chez l'Homme. L'arthrite comporte une inflammation de tissu mou, une formation de pannus et deux ostéopathies distinctes, une résorption ossuse accélérée et une formation anormale d'os périarticulaire. L'étidronate disodique, administré à raison de 4 mg/kg/jour par voie cutanée, à partir de l'injection de l'adjuvant, inhibe nettement la résorption osseuse, la formation de pannus, l'érosion inflammatoire de cartilage et la formation d'os pathologique, en rapport avec l'adjuvant. Après arrêt du traitement à l'étidronate disodique, la formationd'os pathologique devient visible radiographiquement deux semaines après l'arrêt du traitement. Le rôle de l'étidronate disodique sur la régulation de la résorption osseuse et sur la concentration du calcium et du phosphate, dans les tissus environnants, ainsi que les rapports des phénomènes arthritiques avec ce type d'expérience sont envisagés. Les résultats suggèrent la possibilité de l'emploi de l'étidronate disodique dans certaines formes d'arthrites humaines.
    Abstract: Zusammenfassung Freund's Adjuvans, das aus abgetötetem und in Mineralöl suspendiertem Mycobacterium butyricum besteht, verursacht bei Ratten eine arthritische Reaktion, welche gewissen Arthritisformen beim Menschen ähnlich ist. Die arthritische Reaktion besteht in einer Entzündung der Weichteile, einer Pannusbildung und zwei verschiedenen Osteopathien, ferner in einer beschleunigten Knochenresorption und einer abnormen periartikulären Knochenbildung. Wird gleichzeitig mit der Verabreichung des Adjuvans Dinatriumetidronat in einer Dosis von 4 mg/kg/Tag subcutan gegeben, so werden sowohl die Knochenresorption, als auch die Pannusbildung, die entzündliche Knorpelerosion und die mit der Adjuvansgabe verbundene pathologische Knochenbildung merklich gehemmt. Wurde die Dinatriumetidronat-Behandlung abgebrochen, so wurde die pathologische Knochenbildung innerhalb 2 Wochen nach Behandlungsabbruch röntgenologisch sichtbar. Die Kontrollfunktion des Dinatriumetidronates bei der Knochenresorption und bei der Calcium- und Phosphatkonzentration in den umliegenden Geweben, sowie die Beziehung dieser Substanz zu den arthritischen Prozessen, wie sie bei diesem Rattenversuch vorliegen, werden besprochen. Auf Grund dieser Resultate ist eine potentielle Anwendung des Dinatriumetidronates bei gewissen Formen der menschlichen Arthritis denkbar.
    Notes: Abstract Freund's Adjuvant, consisting of deadMycobacterium butyricum suspended in mineral oil, produces an arthritic response in rats which resembles certain forms of arthritis in man. The arthritic response consists of soft tissue inflammation, pannus formation and two separate osteopathies; accelerated bone resorption and abnormal periarticular bone formation. Disodium etidronate administered at 4 mg/kg/day subcutaneously from the time of adjuvant injection markedly inhibited bone resorption, pannus formation, inflammatory erosion of cartilage, and the pathologic bone formation associated with the adjuvant model. When the disodium etidronate treatments were discontinued, the pathologic bone formation became radiologically visible within two weeks after cessation of treatment. The role of disodium etidronate in controlling bone resorption and surrounding tissue concentration of calcium and phosphate and the relation to arthritic processes in this rat model are discussed. The data suggest a potential use of disodium etidronate in some forms of human arthritis.
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    Calcified tissue international 9 (1972), S. 310-324 
    ISSN: 1432-0827
    Keywords: Strontium ; Calcification ; Collagen ; Nucleation ; Exchange
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Une matrice non calcifiée de tendon de bœuf fixe environ 2 μ moles de Sr2+/100 mg de matrice. La fixation de Sr2+ est inhibée par du Mg2+, du Ca2+ et de l'urée, mais n'est pas modifiée par le phosphate inorganique, le phosphonoacétate ou le méthylène-diphosphonate. Le Sr2+ inhibe la calcification réticulaire et cette inhibition est inversée par le Ca2+, mais non par HPO 4 2− . Bien que la formation réticulaire d'apatite de strontium ne soit pas induite par cette matrice, une précipitation élevée de Sr2+ est observée au niveau de la matrice déjà calcifiée. Cette dernière ne nécessite pas l'addition de phosphate. Ce phénomène s'accompagne par une libération presque équimolaire de Ca2+ de la phase soluble et il est inhibé par des composés qui inhibent également les réactions d'échange du45Ca2+ et du32P-HPO 4 2− . Ces résultats semblent indiquer que le Sr2+ peut se fixer à la matrice par un processus sensible au Mg2+ et à l'urée, ainsi que par incorporation à la phase minérale liée à la matrice. Ce dernier mécanisme pourrait se produire par échange isoionique nécessitant l'adjonction de phosphate. Le rapport possible de ces observations avec les diverses étapes de la calcification est envisagé.
    Abstract: Zusammenfassung Nichtverkalkte Matrix aus Rindersehne bindet ungefähr 2 μMol Sr2+/100 mg Matrix. Die Aufnahme von Sr2+ wird durch Mg2+, Ca2+ und Harnstoff gehemmt, jedoch durch anorganisches Phosphat, Phosphonoacetat oder Methylendiphosphonat nicht beeinflußt. Sr2+ hemmt die Netto-Verkalkung, und diese Hemmung wird durch Ca2+, aber nicht durch HPO 4 2− verhindert. Obschon die Netto-Strontiumapatitbildung durch diese Matrix nicht angeregt wird, wurde eine erhöhte Sr2+-Aufnahme durch vorgängig verkalkte Matrix beobachtet. Für diese Aufnahme wird kein zusätzliches Phosphat benötigt; sie wird von einer nahezu äquimolaren Ca2+-Abgabe an die lösliche Phase begleitet und von Substanzen gehemmt, welche auch die45Ca2+- und32P-HPO 4 2− -Austauschreaktionen hemmen. Diese Resultate können in dem Sinne interpretiert werden, daß Sr2+ fähig ist, sich durch einen Mg2+-und Harnstoff-empfindlichen Vorgang, sowie durch Einbau in die Matrix-gebundene Mineralphase an die Matrix zu binden. Es wird vorgeschlagen, daß dieser Einbau durch einen heteroionischen Austauschmechanismus geschieht, welcher mit dem isoionischen Austausch, der zusätzlich Phosphat benötigt, nicht identisch ist. Es wird diskutiert, ob diese Beobachtungen möglicherweise mit einem in mehreren Schritten ablaufenden Verkalkungsprozeß im Zusammenhang stehen.
    Notes: Abstract Uncalcified matrix derived from beef tendon will bind approximately 2 μmoles Sr2+/100 mg matrix. Sr2+ uptake is inhibited by Mg2+, Ca2+ and urea but is not influenced by inorganic phosphate, phosphoacetate, nor methylenediphosphonate. Sr2+ inhibits net calcification and this inhibition is reversed by Ca2+ but not by HPO 4 2− . Although net strontium apatite formation is not induced by this matrix, an elevated Sr2+ uptake by previously calcified matrix was observed. The latter does not require added phosphate, is accompanied by a nearly equimolar release of Ca2+ to the soluble phase and is inhibited by compounds that also inhibit the45Ca2+ and32P-HPO 4 2− exchange reactions. These results are interpreted to indicate that Sr2+ can bind to the matrix by a Mg2+ and urea-sensitive process as well as by incorporation into the matrix-bound mineral phase. It is proposed that the latter occurs by a heteroionic exchange mechanism that is not identical to the isoionic exchange which requires added phosphate. The possible relationship of these observations to a multi-step calcification process is discussed.
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    Calcified tissue international 10 (1972), S. 91-102 
    ISSN: 1432-0827
    Keywords: Hydroxyapatite ; Calcification ; Phases ; Growth ; Microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé La croissance cristalline de l'hydroxyleapatite à 25° et à pH constant de 7.4 a été étudiée à l'aide du microscope électronique à balayage. La technique reproductible de croissance par ensemencement à partir de solutions stables sursaturées est utilisée efficacement pour produire des échantillons de minéral à divers stades distincts de croissance. Des changements de phase sont observés avec le progrès de la croissance; ils correspondent dans le temps avec les résultats cinétiques obtenus antérieurement. Un essai de rationalisation est tenté à la lumière des mécanismes proposés pour la formation d'hydroxyleapatite dans des conditionsin vivo.
    Abstract: Zusammenfassung Das Kristallwachstum von Hydroxyapatit bei 25° und einem konstanten pH von 7,4 wurde mit Hilfe eines Raster-Elektronenmikroskopes studiert. Die reproduzierbare Technik des Keimwachstums aus stabilen übersättigten Lösungen wurde mit Erfolg verwendet, um Mineralproben in verschiedenen bestimmten Stadien des Wachstums zu erhalten. Phasenveränderungen wurden beim fortschreitenden Wachstum beobachtet, und diese stimmten zeitlich gut überein mit kinetischen Resultaten, über welche früher berichtet wurde. Es wurde versucht, diese Beobachtungen zu erklären in Anbetracht von Mechanismen, welche für die Bildung von Hydroxyapatit unterin vivo-Bedingungen vorgeschlagen wurden.
    Notes: Abstract The crystal growth of hydroxyapatite at 25° and at a constant pH of 7.4 has been studied with the aid of a scanning electron microscope. The reproducible technique of seeded growth from stable supersaturated solutions was used effectively to produce samples of the mineral at various distinct stages of growth. Phase changes were observed as the growth proceeded and these corresponded favorably in time with kinetic results reported earlier. An attempt was made to rationalize the observations in light of mechanisms proposed for the formation of hydroxyapatite under conditions foundin vivo.
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