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  • Electron microscopy  (1,196)
  • Rat  (826)
  • Springer  (1,967)
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  • 1
    ISSN: 1432-1351
    Keywords: Key words Olfactory receptor cells ; Olfactory bulbectomy ; Olfactory axotomy ; Electrophysiology ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract This study investigated whether contact with the olfactory bulb was necessary for developing and renewing olfactory receptor neurons (ORNs) to attain normal odorant responsiveness, and whether the anatomical and functional recoveries of the olfactory epithelium were similar in both bulbectomized (BE) and bilaterally axotomized (AX) preparations. In vivo electrophysiological recordings were obtained in response to amino acids, a bile acid [taurolithocholic acid sulfate(TLCS)] and a pheromonal odorant [17α, 20β,-dihydroxy-4-pregnen-3-one (17,20P)] from sexually immature goldfish. Both transmission and scanning electron microscopy indicated that the olfactory epithelium degenerated in BE and AX goldfish. Within 1–2 weeks subsequent to the respective surgeries, responses to high concentrations (〉0.1 mmol · l−1) of the more stimulatory amino acids remained, whereas responses were no longer obtainable to TLCS and 17,20P. At 4 weeks, responses to amino acid stimuli recovered to control levels, while responses to TLCS and 17,20P were minimal. By 7 weeks post bilateral axotomy, the olfactory epithelium recovered to a condition similar to control sensory epithelium; however, the rate of degeneration and proliferation of receptor neurons in BE preparations appeared to remain in balance, thus blocking further recovery of the olfactory epithelium. At 7 weeks post surgery, odorant responses of AX and BE goldfish to TLCS and 17,20P were still recovering.
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  • 2
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    The journal of membrane biology 140 (1994), S. 215-223 
    ISSN: 1432-1424
    Keywords: Insulin receptor ; Membrane reconstitution ; Electron microscopy ; Quaternary structure ; Immunogold labeling
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Insulin receptors were incorporated into liposomes by two different procedures, one using dialysis and one using detergent removal by Bio-Beads. Receptor incorporation was analyzed by gradient centrifugation and electron microscopy. Reconstituted receptors projected up to 12 nm above the membrane and exhibited a T-shaped structure compatible with that previously described for the solubilized receptor. Insulin binding and autophosphorylation experiments indicated that approx. 50% of the receptors were incorporated right-side out. Such random orientation was confirmed by immunogold labeling of the α- and the β-subunit of the receptor. Immunogold labeling of the C-terminus of the β-subunit indicates that it resides about 6 nm off the membrane, while two α-subunit epitopes were labeled at about twice this distance, confirming that the α-subunit is harbored in the cross-bar of the T-structure.
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  • 3
    ISSN: 1432-2021
    Keywords: Key words Cristobalite ; Tridymite ; Phase transformation ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences , Physics
    Notes: Abstract Using minimum exposure techniques, it is feasible to perform high resolution electron microscopy on the α-cristobalite phase of (Si0.9 Ge0.1)O2, which is extremely radiation sensitive. Such images reveal atomic scale information of twins and tridymite-like stacking faults on (1 1 1)β planes, as well as of domain boundaries resulting from the β→α transition. Polytype structures are formed in certain cases. Morphological features suggest that the phase transformation cristobalite → tridymite proceeds by means of a zonal dislocation mediated synchro-shear process on (1 1 1)β planes; the geometry of this process is analyzed.
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  • 4
    ISSN: 1432-2048
    Keywords: Bradyrhizobium ; Electron microscopy ; Glycine (root nodules) ; High-pressure freezing ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract High-pressure freezing of chemically untreated nodules of soybean (Glycine max (L.) Merr.), in sharp contrast to chemical fixation and prefixation, appears to preserve the ultrastructure close to the native state. This is supported by the observation that the peribacteroid membrane of high-pressure-frozen samples is tightly wrapped around the bacteroids, a finding that is fully consistent with the current views on the physiology of oxygen and metabolite transport between plant cytosol and bacteroids. In soybean root nodules, the plant tissue and the enclosed bacteria are so dissimilar that conventional aldehyde-fixation procedures are unable to preserve the overall native ultrastructure. This was demonstrated by high-pressure freezing of nodules that had been pre-fixed in glutaraldehyde at various buffer molalities: no buffer strength tested preserved all ultrastructural aspects that could be seen after high-pressure freezing of chemically untreated nodules.
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  • 5
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    Journal of molecular evolution 24 (1987), S. 260-271 
    ISSN: 1432-1432
    Keywords: Mouse ; Rat ; Two-dimensional electrophoresis ; Quantitative variability of proteins ; Qualitative variability of proteins ; Protein classes ; Membrane proteins ; Organ-specific proteins ; Regulatory genes ; Speciation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Proteins of membranes and cytosols were extracted from the livers and brains of mice (inbred strain DBA/6J) and rats (inbred strain DA/Han) and separated by two-dimensional electrophoresis (2-DE). The 2-DE patterns were compared with regard to qualitative (spot position) and quantitative (spot intensity) characteristics of the proteins of these two species. The following results were obtained: (1) Brain had more (higher percentage) conservative proteins (proteins found in both mice and rats) than liver; (2) plasma membranes had more conservative proteins than the cytosols; (3) organ-unspecific proteins contained more conservative proteins than relatively organ-specific proteins; (4) the pattern of distribution of genetic variability among different classes of proteins represented by findings 1–3 was the same for the qualitative and quantative characteristics of the proteins; and (5) some observations indicated that quantitative variability occurred more frequently among proteins than did qualitative variability. Our conclusion is that regulatory sequences in the DNA (regulatory genes) are subjected to functional constraints that differ in strength among different classes of proteins by the same ratios as the constraints acting on the structural genes. The overall effect of the selective pressure is, however, less stringent for regulatory genes than for structural genes. The results obtained here by comparing two different species are very similar to previous results we obtained by studying different subspecies (inbred strains of the mouse). From this finding arises a new concept: the study of molecular evolution on the basis of different classes of proteins. Our results were compared with data from the literature that were obtained in part from studies on cultured cells. The comparison suggested that cultured cells have lost their tissue-specific proteins, and so generate predominantly extremely conservative proteins.
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  • 6
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    Journal of molecular evolution 26 (1987), S. 347-357 
    ISSN: 1432-1432
    Keywords: Ribosome structure ; Electron microscopy ; Image analysis ; Evolutionary lineages
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Multivariate statistical analysis and classification techniques are powerful tools in sorting noisy electron micrographs of single particles according to their principal features, enabling one to form average images with an enhanced signal-to-noise ratio and a better reproducible resolution. We apply this methodology here to determining the characteristic views of the large (50S) ribosomal subunits from the eubacteriumEscherichia coli and the archaebacteriaMethanococcus vannielii, Sulfolobus solfataricus, andHalobacterium marismortui. Average images were obtained of the subunit in the common crown and kidney projections, but views of the particle in orientations intermediate between these two extremes were also elucidated for all species. These averages show reproducible detail of up to 2.0 nm resolution, thus enabling the visualization and interspecies comparison of many structural features as a first step toward comparing the actual three-dimensional structures. Our results disprove evolutionary lineages recently postulated on the basis of electron microscopical images of ribosomal subunits.
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  • 7
    ISSN: 1432-1432
    Keywords: Carcinoembryonic antigen ; Evolution ; Gene family ; Human ; Rat ; Synonymous substitutions ; Silent molecular clock ; Evolutionary trees
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Various rodent and primate DNAs exhibit a stronger intra- than interspecies cross-hybridization with probes derived from the N-terminal domain exons of human and rat carcinoembryonic antigen (CEA)-like genes. Southern analyses also reveal that the human and rat CEA gene families are of similar complexity. We counted at least 10 different genes per human haploid genome. In the rat, approximately seven to nine different N-terminal domain exons that presumably represent different genes appear to be present. We were able to assign the corresponding genomic restriction endonuclease fragments to already isolated CEA gene family members of both human and rat. Highly similar subgroups, as found within the human CEA gene family, seem to be absent from the rat genome. Hybridization with an intron probe from the human nonspecific cross-reacting antigen (NCA) gene and analysis of DNA sequence data indicate the conservation of noncoding regions among CEA-like genes within primates, implicating that whole gene units may have been duplicated. With the help of a computer program and by calculating the rate of synonymous substitutions, evolutionary trees have been derived. From this, we propose that an independent parallel evolution, leading to different CEA gene families, must have taken place in, at least, the primate and rodent orders.
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  • 8
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    Machine vision and applications 4 (1991), S. 271-285 
    ISSN: 1432-1769
    Keywords: Electron microscopy ; three-dimensional vision ; surface reconstruction ; stereo ; shape from shading ; dynamic programming
    Source: Springer Online Journal Archives 1860-2000
    Topics: Computer Science
    Notes: Abstract The computational reconstruction of surface topographies from scanning electron microscope (SEM) images has been extensively investigated in the past, but fundamental image processing problems still exist. Since conventional approaches adapted from general-purpose image processing have not sufficiently met the requirements in terms of resolution and reliability, we have explored combining different methods to obtain better results. This paper presents a least-squares combination of conventional stereoscopy with “shape from shading” and a way of obtaining self-consistent surface profiles from stereoscopy and “stereo-intrinsic shape from shading” using dynamic programming techniques. Results are presented showing how this combined analysis of multi-sensorial data yields improvements of the reconstructed surface topography that cannot be obtained from individual sensor signals alone.
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  • 9
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    Journal of comparative physiology 170 (1992), S. 645-650 
    ISSN: 1432-1351
    Keywords: Milk ejection reflex ; Positive pressure oscillations ; Motor activity of pups ; Tactile stimulation of nipples ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Changes of positive pressure exerted by pups on nipples during sucking were investigated using anesthetized, lactating dams. It was found that, every 50–60 s, individual pups performed bouts of pressure oscillations (3/s) of high amplitude which lasted about 10–12 s and coincided with periods of increased motor activity. During the intervals, when pups were quiet, series of low-amplitude oscillations (3/s) were also observed. Using a strain measuring method to record the activity of sucking pups, synchronization of activity of two or more pups was found to occur periodically every 25–30 s and, most frequently, 10–30 s before the reflex increase of milk pressure. In further experiments, artificial tactile stimulation was applied to the dam's nipples using the joint action of suction and positive pressure. Following a short-term (10–20 s) increase in frequency and amplitude of artificial nipple stimulation, 60%–80% of all reflexive peaks of milk pressure were elicited with a latency of 19 ± 5 s. This suggests that there are specific conditions under which the stimulation of nipples by pups may trigger the formation of the milk ejection reflex in the rat.
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  • 10
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    Journal of comparative physiology 171 (1992), S. 657-664 
    ISSN: 1432-1351
    Keywords: Inferior olivary complex ; Cerebellum ; 3-acetylpyridine ; Harmaline ; Learning ; Memory ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary DA/HAN-strained male rats (pigmented rats) were submitted to two experimental tasks consisting of spatial learning (water-escape) and a passive avoidance conditioning. Both these tasks were performed by different animals. In order to destroy the inferior olivary complex, the animals were injected with 3-acetylpyridine either 9 days prior to the initial learning session or 24 h after completion of the learning task. They were retested (retrieval test) 10 days after the initial learning was achieved. Learning and retention were compared to those noted in control rats. Administration of 3-acetylpyridine before the initial learning did not prevent the spatial learning but the scores were greatly altered and the number of trials needed to reach the fixed learning criterion was much greater than in controls. However, 10 days later the animals had memorized their initial experience. Injection of 3-acetylpyridine after the initial learning session impaired memory: the animals had completely forgotten their initial learning. It can therefore be concluded that lesion of the afferent climbing fibres to the cerebellar cortex alters learning and retention of a spatial task. Such a lesion does not interfere with learning and retention of a passive avoidance conditioning, since in this condition the experimental animals injected with 3-acetylpyridine either before or after the initial learning behave similarly to controls. The effects of the inferior olivary complex lesion are obviously different according to the task to be learnt, suggesting that these two tasks do not require the integrity of the same nervous structures.
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  • 11
    ISSN: 1432-1351
    Keywords: Rat ; Melatonin ; Circadian rhythm ; 5-hydroxytryptophan
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The rhythm in melatonin production in the rat is driven by a circadian rhythm in the pineal N-acetyltransferase (NAT) activity. Rats adapted to an artificial lighting regime of 12 h of light and 12 h of darkness per day were exposed to an 8-h advance of the light-dark regime accomplished by the shortening of one dark period; the effect of melatonin, triazolam and fluoxetine, together with 5-hydroxytryptophan, on the reentrainment of the NAT rhythm was studied. In control rats, the NAT rhythm was abolished during the first 3 cycles following the advance shift. It reappeared during the 4th cycle; however, the phase relationship between the evening rise in activity and the morning decline was still compressed. Melatonin accelerated the NAT rhythm reentrainment. In rats treated chronically with melatonin at the new dark onset, the rhythm had already reappeared during the 3rd cycle, in the middle of the advanced night, and during the 4th cycle, the phase relationship between the evening onset and the morning decline of the NAT activity was the same as before the advance shift. In rats treated chronically with melatonin at the old dark onset or in those treated with melatonin 8 h, 5 h and 2 h after the new dark onset during the 1st, 2nd and 3rd cycle, respectively, following the advance shift, the NAT rhythm reappeared during the 3rd cycle as well but in the last third of the advanced night only. Neither triazolam nor fluoxetine together with 5-hydroxytryptophan administered around the new dark onset facilitated NAT rhythm reentrainment after the 8-h advance of the light-dark cycle.
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  • 12
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    Journal of molecular evolution 33 (1991), S. 442-449 
    ISSN: 1432-1432
    Keywords: Humans ; Mouse ; Rat ; Codon usage ; Mutation bias ; Selection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A new statistical test has been developed to detect selection on silent sites. This test compares the codon usage within a gene and thus does not require knowledge of which genes are under the greatest selection, that there exist common trends in codon usage across genes, or that genes have the same mutation pattern. It also controls for mutational biases that might be introduced by the adjacent bases. The test was applied to 62 mammalian sequences, the significant codon usage biases were detected in all three species examined (humans, rats, and mice). However, these biases appear not to be the consequence of selection, but of the first base pair in the codon influencing the mutation pattern at the third position.
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  • 13
    ISSN: 1432-0827
    Keywords: Rat ; Incisor ; Amelogenesis ; Acid phosphatase ; Ferritin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Acid phosphatase was localized in rat incisor ameloblasts without prior decalcification. Whenβ-glycerophosphate was used as the substrate, an intense reaction was observed in the supranuclear region of the secretory ameloblasts. But the reaction was dramatically reduced at the transitional stage and was very weak in the maturation ameloblasts. Whenp-nitrophenylphosphate was the substrate, the reaction product was consistently seen in the Golgi cisternae and the vesicular components of the ameloblasts at all stages of enamel development. These observations suggest that there are two acid phosphatases in ameloblasts. One is in the secretory ameloblasts and the other in the transition and maturation ameloblasts. X-ray micro-analyses for Fe and Pb showed that Fe and acid phosphatase were in the ferritin-containing vesicles at the later stage of enamel maturation. This evidence suggests that ferritin is digested in these vesicles for the release of the Fe pigment to the enamel. An increase in the number of intercellular bridges between ameloblasts was correlated with the dramatic decrease in height of ameloblasts at the pigment release stage. The ameloblast membranes were acid phosphatase positive at the intercellular bridges whenp-nitrophenylphosphate was the substrate. This activity may be involved in the reduction in the surface area of the ameloblast membranes.
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  • 14
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    Calcified tissue international 33 (1981), S. 119-124 
    ISSN: 1432-0827
    Keywords: Avian eggshell ; Microstructure ; Electron microscopy ; Electron diffraction ; Calcite growth
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The eggshell of the domestic fowl has been studied by transmission electron microscopy and diffraction. Thin sections of shell were prepared by chemical and ion-beam thinning techniques. Each calcite column of the palisade layer consisted of crystallites of diameter 20 to 30 µm with some tendency for crystallite alignment within a single column. Evidence indicates that there was no significant preferred orientation in the palisade layer as a whole. Only in the surface layer was any preferred orientation detected, and here {1014} planes tended to lie parallel to the surface. The results are compared with previously published data, and calcite nucleation and growth are discussed.
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  • 15
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    Calcified tissue international 10 (1972), S. 238-251 
    ISSN: 1432-0827
    Keywords: Growth ; Bone ; Rat ; Tetracycline
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé La vitesse de croissance normale en longueur de la métaphyse proximale du tibia est déterminée chez le rat Sprague-Dawley entre les âges de 20 et 100 jours, en utilisant la méthode à la tétracycline. Le taux de croissance ne varie que légèrement dans les groupes d'âges différents. Il est plus élevé chez les animaux jeunes et décroit considérablement en fonction de l'augmentation de l'âge. Les rats mâles présentent une croissance plus élevée que les femelles. Cette étude a pour but de mettre au point une méthode permettant de déterminer les facteurs expérimentaux, liés à la croissance en longueur du rat.
    Abstract: Zusammenfassung Die normale Längenwachstums-Geschwindigkeit der proximalen Wachstumsplatte der Tibia wurde bei Sprague-Dawley-Ratten in einem Alter zwischen 20 und 100 Tagen mittels der Tetracyclinmethode gemessen. Die Wachstumsgeschwindigkeit variierte nur wenig innerhalb der einzelnen Altersgruppen. Die Geschwindigkeit war bei jungen Tieren am höchsten und nahm mit zunehmendem Alter beträchtlich ab. Männliche Ratten wuchsen schneller als weibliche. Diese Arbeit dient als Grundlage, um die experimentelle Beeinflussung des Längenwachstums der Ratte abschätzen zu können.
    Notes: Abstract The rate of normal growth in length from the proximal growth plate of the tibia in the Sprague-Dawley rat was measured between 20 and 100 days of age using the tetracycline method. The growth rate varied only slightly within different age groups. The rate was highest in young animals and decreased considerably with increasing age. Male rats grew faster than female. This study is intended to provide a base for an evaluation of experimental influence on the growth in length of the rat.
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  • 16
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    Calcified tissue international 13 (1973), S. 113-129 
    ISSN: 1432-0827
    Keywords: Kinetics ; Growth ; Epiphyses ; Hypophysectomy ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé La largeur de la métaphyse tibiale, la zone indifférenciée, la zone sériée et les cellules en dégénerescence ont été observées chez des rats Sprague-Dawley normaux et hypophysectomisés. La production cellulaire de la métaphyse est déterminée sur la base de la croissance osseuse longitudinale déterminée par l'oxytétracycline et la taille des cellules en dégénérescence. La diminution de la croissance osseuse longitudinale, en fonction de l'augmentation de l'âge et après hypophysectomie, est due partiellement, à la diminution de production cellulaire et partiellement à une décroissance de la taille des cellules en dégénérescence dans la métaphyse. L'influence de la production cellulaire et de l'activité mitotique prédomine.
    Abstract: Zusammenfassung Die Breite der proximalen Wachstumsplatte der Tibia, deren undifferenzierter und säulenförmiger Zone und die Größe der nahe bei der Metaphyse auftretenden degenerativen Zellen wurden in normalen und hypophysektomierten Sprague-Dawley-Ratten bestimmt. Die Zellproduktion in der Wachstumsplatte wurde aus dem longitudinalen Knochenwachstum berechnet, welches mittels Oxytetracyclin und der Größe der degenerativen Zellen bestimmt wurde. Es wurde festgestellt, daß die Abnahme des longitudinalen Knochenwachstums bei zunehmendem Alter und nach Hypophysektomie zum Teil einem Rückgang in der Zellproduktion, zum Teil einer Verminderung der Größe der degenerativen Zellen in der Wachstumsplatte zuzuschreiben ist. Der Einfluß der Zellproduktion, und somit der mitotischen Aktivität, herrscht vor.
    Notes: Abstract The width of the proximal growth plate of the tibia, its undifferentiated and columnar zone and the size of the degenerative cell close to the metaphysis, were determined in normal and hypophysectomized Sprague-Dawley rats. The cell production in the growth plate was calculated from the longitudinal bone growth determined with oxytetracycline and the degenerative cell size. It was found that the decrease in longitudinal bone growth with increasing age and after hypophysectomy, is due partly to a decrease in cell production, and partly to a decrease in degenerative cell size in the growth plate. The influence of cell production and thus the mitotic activity predominates.
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  • 17
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    Calcified tissue international 22 (1977), S. 77-84 
    ISSN: 1432-0827
    Keywords: Fluoride ; Blood plasma ; Enamel mineralization ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract It is postulated that tissue fluid F concentrations are the primary determinants of fluoride effects on bones and developing teeth and that these concentrations are dependent on, or mirrored by, blood plasma F. It has earlier been shown that the plasma F levels are dependent on the dietary F supply as well as on skeletal F concentration. Fasting and post-ingestion or postinjection plasma F levels have been determined in rats on F doses that cause different degrees of enamel fluorosis. The results indicate that temporary peak values rather than elevated fasting values are responsible for the occurrence of enamel fluorosis and that the peak values must approach about 10 μM in order to block enamel formation by the ameloblasts. The diagnostic and prognostic importance of plasma F determinations is discussed.
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  • 18
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    Calcified tissue international 3 (1969), S. 96-99 
    ISSN: 1432-0827
    Keywords: Calcium metabolism ; Strontium metabolism ; Fracture ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé La relation entre la résorption du85Sr et du47Ca était mesuré au tibia et fémur des rats 24 heures après l'injection. La différence entre les manches et les bouts était, remarquable, mais pas entre les os séparates; en les tibiae, qui etait fracturé depuis 7 semaines et guéries, la relation etait identique à celui des tibiae manches normals. On a présumé que la découverte était due à des différences qualitatives plustot que quantitatives entre les procédé de minéralisation dans l'os cortical et trabeculair.
    Abstract: Zusammenfassung Das Verhältnis zwischen der Abnahme von85Sr und47Ca wurde von der Tibia und Femur 24 Std nach der Injektion bei Ratten gemessen. Man fand einen signifikanten Unterschied zwischen den Schaften und den Enden, aber nicht zwischen den verschiedenen Knochen. In 7 Wochen alten, verheilten Tibiafrakturen war das Verhältnis genau so wie in einem normalen Tibiaschaft. Das Resultat berechtigte zu der Annahme, daß der Unterschied in dem Mineralisierungsprozeß zwischen corticalen und spongiösen Knochen wahrscheinlich qualitativ und nicht quantitativ ist.
    Notes: Abstract The ratio between the uptake of85Sr and47Ca was measured in the tibiae and femora of rats 24 h after injection of the tracers. There was a significant difference between shafts and ends but not between the different bones; in healed tibial fractures, 7 weeks old, the ratio was identical to that of normal tibial shafts. The findings were interpreted to be related to qualitative rather than quantitative differences in mineralization between cortical and trabecular bone.
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  • 19
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    Calcified tissue international 25 (1978), S. 179-190 
    ISSN: 1432-0827
    Keywords: Decalcification ; Electron microscopy ; Calcified matrices
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The ultrastructure of calcifying cartilage and bone has been examined under the electron microscope after using three different methods of decalcification. The first was carried out before embedding (by soaking specimens in EDTA or formic acid), the second after embedding (by floating ultrathin sections on formic acid), and the third after embedding (by soaking embedded specimens in EDTA or formic acid), and with later re-embedding. The first procedure invariably induces drastic changes in the fine structure of the cells and calcified matrix, probably as a results of the extraction of organic material along with extraction of mineral. The second and third procedures make it possible to preserve ultrastructural details perfectly in both cells and calcified matrix. Of the two, the third procedure is preferable because of its greater simplicity. In areas that are still calcifying, these post-embedding decalcification techniques reveal the presence of crystal-associated, filamentous organic structures which are not recognizable in specimens decalcified before embedding. These structures, which could have a key role in inducing and regulating crystal formation and growth, are less evident in fully calcified areas (but not at their borders). This may partly be due to the loss of glycan components in the matrix during calcification. The most important determinant, however, seems to be the fact that during calcification the components of the matrix, including collagen fibrils, are involved in an aggregation process which reduces the amounts of free chemical groups available for reaction with the stain solution. Because post-embedding decalcification does not disturb this state of aggregation, the stainability of the matrix and the electron microscopic evidence of its components remain very low.
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  • 20
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    Calcified tissue international 23 (1977), S. 215-223 
    ISSN: 1432-0827
    Keywords: Amorphous mineral ; Bone ; Electron microscopy ; Ultracryotomy ; Ultramicro-incineration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The fine structure of the extracellular phase of avian medullary bone and embryonic chick femur was examined in thin sections prepared by ultracryotomy and ultramicroincineration. Since contact with solutions was completely avoided, little or no loss or dislocation of mineral constituents could occur. Amorphous bone mineral (ABM) was present in two forms: as 15–30 nm spheres and as a structure-free haze. Removal of all organic material by low temperature ashing left the ABM intact. Crystals were usually associated with the ABM. In newly ossifying regions clusters or nodules of randomly oriented crystals and ABM appeared to coalesce when they reached approximately 1 μm in diameter. In highly calcified regions crystals appeared to be oriented along collagen fibers. ABM did not appear to be associated with collagen. Unmineralized collagen was visible in osteoid after staining with dry OsO4 vapor and it appeared to be diverted around nodules. Structures which resembled matrix vesicles were present. Selected area electron diffraction patterns indicated the presence of hydroxyapatite.
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  • 21
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    Calcified tissue international 30 (1980), S. 43-50 
    ISSN: 1432-0827
    Keywords: Osteogenesis ; In vitro ; Electron microscopy ; Mineralization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Chick limb mesenchymal cells differentiate into muscle, cartilage, fibrous, and bone tissue. Previous reports show that when stage 24 limb mesenchymal cells are cultured in vitro, chondrocytes, myocytes, fibrocytes, and osteoblasts can be identified on the basis of morphological and biochemical parameters. The study reported here demonstrates that phenotypic expression in culture seems to be dependent on the initial plating density, Scanning electron microscopic observations indicate that when stage 24 limb mesenchymal cells are initially seeded at high densities (5 × 106 cells per 35 mm culture dish), mounds of cells appear in culture. These mounds represent cartilage nodules composed of a fine fibrous matrix and chondrocytes, surrounded by a loose fibrous connective tissue matrix. Cultures initially plated at intermediate densities (2.0–2.5 × 106 cells/35 mm culture dish) produce a flattened layer of fibrocytes overlying a matrix of collagen fibers and calcium phosphate deposits as determined by electron-microprobe analysis; these observations are indicative of osteoblast expression. Cells seeded at this intermediate density appear larger and possess greater surface area than cells seeded at high density. It is suggested that conditions that permit such increased cell surface area coupled with a relative compaction due to cell crowding may provide conditions permissive for osteogenesis. Based on morphological criteria, it appears that chick limb mesenchymal cell osteogenesis in vitro is not associated with chondrogenesis but represents a separate route of phenotypic expression.
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  • 22
    ISSN: 1432-0827
    Keywords: Parathyroid hormone ; Osteoclasts ; Electron microscopy ; Morphometry ; Metaphysis
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Summary The effects of parathyroid hormone (PTH) on the size of the osteoclasts, nuclei, ruffled borders, and clear zones in long bones of thyroparathyroidectomized (TPTX) rats were quantitated as a function of time. These data were compared with the number of osteoclasts in the bone and with plasma calcium levels. A significant increase in the average size of the ruffled borders was demonstrated 30 min after injection of 50 U of purified bovine PTH, and of the clear zones 30–90 min after PTH. This was followed at 90 min by an increase in the average size of the cells. The sizes of ruffled borders and clear zones dropped sharply to control levels after 6 h, whereas the size of the cells remained elevated up to 12 h and returned to control values at 24 h. Plasma calcium levels were increased, but not significantly, between 30 min and 6 h. An increase in the number of osteoclasts was significant after 12 h. Removal of the parathyroid glands did not diminish the normal activity of osteoclasts. In animals with intact glands injection of 50 U of PTH did not cause a significant change in cell size or resorbing apparatus. It is concluded that PTH acts to rapidly stimulate the bone resorptive activity of osteoclasts and to cause a delayed increase in their number.
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  • 23
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    Calcified tissue international 32 (1980), S. 63-68 
    ISSN: 1432-0827
    Keywords: Rat ; Bone formation ; Fluorochrome ; Microphotometry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary A method for quantitative studies of the formation rate of bone has been developed. After vital staining with calcein, the fluorescence of a bone section was measured with a microphotometer controlled by a mini computer. After staining the bone structure with alizarin red S in a second step, the section was measured in transmitted light. The two data sets were combined and the shortest distances between the bone surface and the fluorescence lines were computed. With this information the distance distribution and the bone area between the label and the surface could be calculated in two different ways: with the single labeling and the continuous labeling techniques. The advantages and disadvantages of the two methods are discussed and compared with those of other techniques.
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  • 24
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    Calcified tissue international 32 (1980), S. 113-122 
    ISSN: 1432-0827
    Keywords: Rat ; Bone ; Metaphysis ; Quantitative ; Aging
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The purpose of this work was to analyze the proximal tibial metaphysis of the 170 g rat in a quantitative histologic fashion which would allow some relation to tissue age to be established. Stained 3 µm thick tissue sections were analyzed with the aid of a Merz grid on an eyepiece reticule and a light microscope. Tissue mass and cell distribution were studied in all areas. The rate of change in tissue mass during aging of the metaphysis was calculated. Two regions of the metaphysis were identified. One, corresponding to the primary spongiosa, less than 4.45 days of age, is a region of high turnover of hard tissue and high numbers of osteoblasts and osteoprogenitor cells. The other, corresponding to the secondary spongiosa, is a region of relatively low net tissue turnover and low numbers of osteoblasts and osteoprogenitor cells. Osteoclasts were found relatively more uniformly distributed through the metaphysis than were osteoblasts and osteoprogenitor cells. The rate of bone formation in the primary spongiosa is 50 times that found in the Haversian bone of the rib of 5-year-old humans and about 500 times that found at the cortical-endosteal surface of ribs of 5-year-old humans. It is argued that both cell distribution and tissue distribution in the metaphysis support the concept that osteoblasts and osteoclasts, rather than osteocytes, are responsible for the maturation of the metaphysis. The inhomogeneous distribution of both cells and tissue in the metaphysis has definite meaning for the interpretation of findings concerning the incorporation of radionuclides into the skeleton.
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  • 25
    ISSN: 1432-0827
    Keywords: Rat ; Hypophysectomy ; Dietary phosphorus deprivation
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    Notes: Summary The demands of growth are known to exacerbate the effect of phosphorus deprivation (PD). We examined whether changes associated with PD could be prevented in young rats in which growth and growth hormone (GH) were eliminated by hypophysectomy (HPX) and whether PD in normal intact rats (INT) was associated with increased secretion of GH. INT or thyroxine- and ACTH-replaced HPX rats were fed one of the three diets: 0.31% P (NP); 0.027% P (LP), and 0.31% P, pair-fed with LP-mates (NP-PF). The results indicate that HPX did not qualitatively alter several physiologic responses to PD: (a) serum and urinary phosphorus (P) decreased and urinary calcium (Ca) increased; (b) net intestinal Ca retention fell and duodenal sac uptake of45Ca rose; and (c) external P balance was restored and duodenal sac uptake of32P-phosphate increased. Only the hypercalcemia seen in INT, LP rats was prevented by HPX. In INT rats serum immunoassayable GH levels, measured in single samples, were not different between different dietary groups while pituitary bioassayable GH was reduced in both LP and NP-PF rats when compared to the NP rats. Thus, except for hypercalcemia, the physiologic responses associated with PD are not prevented by the elimination of growth and GH, and the development of these responses in INT rats was not associated with a consistent or specific alteration in GH secretion.
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  • 26
    ISSN: 1432-0827
    Keywords: Decalcification ; Electron microscopy ; Bone matrix ; Bone glycoproteins
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    Notes: Summary A solution of triethylammonium EDTA in 80% ethanol was evaluated as a demineralizing reagent for bone in comparison with aqueous solutions of EDTA. Biochemical analysis and acrylamide gel electrophoresis of extracts of finely powdered bovine bone showed that most of the macromolecular components of the organic matrix extractable in aqueous EDTA were retained when the triethylammonium EDTA reagent was used. Ultrastructural examination of chick tibias decalcified with the reagents showed a better preservation of cellular morphology, especially the membranous components, and more uniformly distributed ground substance, though slightly less in quantity, when the aqueous reagent was used. Use of the two reagents appears to be complementary, the alkylammonium reagent being more appropriate for use in studies of the organic matrix of bone, including immunohistochemical studies of bone glycoproteins. The aqueous reagent is more appropriate for use in studies of cellular ultrastructure.
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  • 27
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    Calcified tissue international 34 (1982), S. 376-381 
    ISSN: 1432-0827
    Keywords: Matrix vesicles ; Bone ; Actin ; Rat
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    Notes: Summary Preliminary indications of the occurrence of actin and myosin in crude matrix vesicle preparations have been reported previously. In the present study extracellular matrix vesicles from rat alveolar bone were isolated. They were further purified by a sucrose density gradient. SDS-polyacrylamide gel electrophoresis of the purified vesicles revealed the presence of a polypeptide with a molecular weight of 43 K daltons and with electrophoretic mobility identical to that of blood platelet actin. The limited proteolysis of both 43 K dalton vesicular polypeptide and actin byStaphylococcus aureus-V8-protease revealed three fragments with identical electrophoretic mobility. In addition, the vesicular preparations inhibited the activity of DNase I, a property typical of actin monomers. Filamentous material extracted from matrix vesicles showed ultrastructural features of F-actin. Reaction of this material with heavy meromyosin resulted in arrowhead formation, which is characteristic of acto-heavy meromyosin. The occurrence of actin in extracellular matrix vesicles may account for their budding from the osteoblastic plasma membrane, their possible motility in the matrix, and maintenance of the spherical shape.
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  • 28
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    Calcified tissue international 35 (1983), S. 107-110 
    ISSN: 1432-0827
    Keywords: Calcium ; Glucocorticoid ; Vitamin D ; Osteoporosis ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Eighty-eight adult male Sprague-Dawley rats were given a diet with either (a) 0.5% Ca and 0.6% P or (b) 0.01% Ca and 0.6% P. Osteopenia was created by adding prednisolone to the diet. The prophylactic effect of oral 1,25(OH)2D3 on the osteopenia was studied. It was found that prednisolone osteopenia in the rat was associated with defective Ca absorption. By giving an oral dose of 1,25(OH)2D3, it was possible to maintain normal Ca absorption during prednisolone treatment and to prevent the bone loss. No significant hypercalcemia or any kidney calcifications were seen. These results are in contrast to earlier findings, in which subcutaneous administration of 1,25(OH)2D3 failed to prevent prednisolone osteopenia because of its tendency to increase bone resorption.
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  • 29
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    Calcified tissue international 52 (1993), S. 361-364 
    ISSN: 1432-0827
    Keywords: Calcitonin ; Sustained release ; Copolymer depot ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Studies were carried out to determine whether monolithic depot formulations, prepared using lactide:glycolide copolymers, could be used to administer salmon calcitonin (sCT) to rats in vivo. Formulations containing 2, 5, or 10% (w/w) sCT were administered subcutaneously to female Wistar strain rats. Release of sCT was determined by measurement of peptide in plasma using a specific radioimmunoassay and by measurement of residual sCT in the depots after recovery at postmortem. Plasma calcium concentrations and cumulative weight gain of the animals were used to measure pharmacological effects of the released sCT. Release of sCT from the depots was controlled by the copolymer and was sustained for periods up to 10 days. However, the release of sCT from the depots did not significantly alter plasma calcium concentrations, and effects on cumulative weight gain were small and transient. Peptide loading of the formulations was shown to modify sCT release. Maximal release of sCT from depots containing 10% peptide occurred over a 7 to 14-day period postadministration, with 5% sCT release occurred between days 11 and 14, and with 2% sCT, the period of maximal release was between days 11 and 18. Release of peptide from the depots was essentially complete by 21 days postadministration irrespective of the peptide loading. These data suggest that lactide:glycolide copolymer depots may have application for the convenient clinical administration of sCT in metabolic bone diseases.
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  • 30
    ISSN: 1432-0827
    Keywords: Hyperthyroidism ; Osteopenia ; Bisphosphonate ; Rat
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    Notes: Summary Hyperthyroidism, either endogenous or iatrogenic, leads to increased bone turnover and osteopenia. This study was conducted to examine (1) whether thyroid hormone excess in rats causes bone changes similar to those seen in patients with hyperthyroidism, and (2) the effects of the aminobisphosphonate alendronate on the thyroid hormone-induced bone changes. Sprague-Dawley male rats, divided into four groups, received L-thyroxine (T4) 250 μg/kg/day (+T4) or vehicle (-T4) subcutaneously six times per week and alendronate 1.75 mg/kg (+ALN) or vehicle (-ALN) orally twice a week. Rats were sacrificed after 3 weeks of treatment, blood samples were analyzed for serum T4, triiodo-L-thyronine (T3), and osteocalcin, and the proximal tibiae were processed for histomorphometric analysis. Serum T4 and T3 levels measured 20–24 hours after the last injection were 2 to 2.5-fold higher in +T4 groups than in-T4 groups. Serum osteocalcin was significantly (P 〈 0.05) higher in +T4/-ALN group than in the other groups, which were not statistically different from each other. T4 treatment (+T4/-ALN) significantly decreased the amount of cancellous bone volume (-45%) and increased osteoid surface (+254%), osteoblast surface (+111%), and osteoclast surface (+176%) relative to control values. Alendronate increased the bone volume above control values in both T4-treated (+ T4/ +ALN) and untreated (-T4/ +ALN) rats, and prevented the T4-induced increase in bone turnover in +T4/+ALN rats. It is concluded that (1) excess thyroid hormone induces cancellous bone loss associated with high bone turnover in the rat, and (2) this bone loss can be prevented by alendronate through the inhibition of osteoclastic activity.
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    Calcified tissue international 56 (1995), S. 42-48 
    ISSN: 1432-0827
    Keywords: Phenytoin ; Bone formation ; Osteocalcin ; Alkaline phosphatase ; Osteogenesis ; Rat
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    Topics: Biology , Medicine , Physics
    Notes: Abstract Long-term use of phenytoin for the treatment of epilepsy has been associated with increased thickness of craniofacial bones. The aim of the present study was to evaluate the possibility that low doses of phenytoin are osteogenic in vivo by measuring the effects of phenytoin administration on serum and bone histomorphometric parameters of bone formation in two rat experiments. In the first experiment, four groups of adult male Sprague-Dawley rats received daily I.P. injections of 0, 5, 50, or 150 mg/kg/day of phenytoin, respectively, for 47 days. Serum alkaline phosphatase (ALP) and osteocalcin were increased by 5 and 50 mg/kg/day phenytoin. The increases in osteocalcin and ALP occurred by day 7 and day 21, respectively. The tibial diaphyseal mineral apposition rate (MAR) at sacrifice (day 48) was significantly increased in rats receiving 5 mg/kg/day phenytoin. At a dose of 150 mg/kg/day, the increase in serum ALP, osteocalcin and MAR was reversed. No significant differences in serum calcium, phosphorus, or 1,25(OH)2D3 levels were seen. In a second experiment, three groups of rats received daily I.P. injection of lower doses of phenytoin (i.e., 0, 1, or 5 mg/kg/day, respectively) for 42 days. Phenytoin also did not affect the growth rate or serum calcium, phosphorus, and 25(OH)D3 levels. Daily injection of 5 mg/kg/day phenytoin significantly increased several measures of bone formation, i.e., serum ALP and osteocalcin, bone ALP, periosteal MAR, and trabecular bone volume. However, rats receiving lower doses of phenytoin (i.e., 1 mg/kg/day) did not show significant increases in the serum bone formation parameters. In contrast, metaphyseal osteoblast surface, osteoblast number, osteoid thickness, surface, and volume were all significantly increased in rats treated in 1 mg/kg/day but not with 5 mg/kg/day phenytoin, suggesting that the tibial diaphysis and metaphysis bone formation parameters might have different dose-dependent responses to phenytoin treatment. Administration of the test doses of phenytoin did not significantly affect the histomorphometric bone resorption parameters. In conclusion, these findings represent the first in vivo evidence that phenytoin at low doses (i.e., between 1 and 5 mg/kg/day) is an osteogenic agent in the rat.
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    Calcified tissue international 56 (1995), S. 83-87 
    ISSN: 1432-0827
    Keywords: Immunoradiometric assay ; Parathyroid hormone ; Rat
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    Topics: Biology , Medicine , Physics
    Notes: Abstract Measurement of parathyroid hormone (PTH) in the rat is most often performed with competitive ligand radioimmunoassays (RIA) utilizing heterologous antibodies. We report here the validation of a newly developed homologous immunoradiometric assay (IRMA) for rat PTH. Two different goat antibodies to the amino-terminal sequence of rat PTH are utilized; one is immobilized onto plastic beads to capture the PTH molecules and the other is radiolabeled for detection. To test this new IRMA, 30 Sprague-Dawley rats were randomized into three treatment groups to receive by intraperitoneal injection: (1) saline 1 ml/kg (control); (2) calcium chloride 40 mg/kg (hypercalcemic); and (3) EDTA 300 mg/kg (hypocalcemic). Blood samples were taken at 0, 30, 60, 180, and 300 minutes after administration of the assigned treatment for measurement of ionized calcium (Ca2+) and serum PTH. Most of the variance in PTH levels was found to be due to changes in Ca2+ (r2=0.780, P〈0.0001). There was also a close temporal relationship between the two, with the highest levels of PTH occurring at the same measured time points as the lowest Ca2+, and vice versa. The measured detection limit of the IRMA was 3 pg/ml with intra-and interassay coefficients of variation of 1.74% and 3.07%, respectively. Serial dilutions with pooled rat serum, synthetic rat PTH-(1–34), and synthetic human PTH-(1–34) showed good parallelism with increased specificity for the pooled and synthetic PTH, despite a degree of crossreactivity with hPTH. The assay is able to quantitate rapid changes in PTH, providing all the advantages of IRMA methodology including technical simplicity and speed of performance, and is likely to become a useful tool in investigations of bone, mineral, and renal homeostasis using the rat.
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    Calcified tissue international 8 (1971), S. 165-171 
    ISSN: 1432-0827
    Keywords: Bone ; Ceramic ; Tetracycline ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Un implant céramique non poreux est testé au niveau du fémur de rat en ce qui concerne son adhésivité à l'os. Un certain nombre de techniques morphologiques sont utilisées pour examiner le rapport entre l'implant et l'os néoformé. La microscopie électronique par transmission et la microscopie par fluorescence après marquage à la tétracycline ont donné les meilleurs résultats. Un rapport étroit entre l'os minéralisé et la céramique a été noté en microscopie électronique. Par marquage à la tétracycline, il semble que l'implant puisse stimuler la formation osseuse.
    Abstract: Zusammenfassung Ein unporöses keramisches Implantat in Rattenfemora wurde auf seine Fähigkeit geprüft, sich mit Knochen zu binden. Eine Anzahl morphologischer Techniken wurde verwendet, um die Beziehung zwischen den Oberflächen von Implantat und neuem Knochen zu untersuchen. Transmissions-Elektronenmikroskopie und Fluoreszenzmikroskopie nach Tetracyclinmarkierung waren die erfolgreichsten Techniken. Eine enge Beziehung zwischen mineralisiertem Knochen und dem Keramikimplantat konnte mit der Transmissions-Elektronenmikroskopie nachgewiesen werden. Das Aussehen der Tetracyclinmarkierung im keramischen Implantat deutet darauf hin, daß dieses wahrscheinlich die Fähighkeit hat, Knochenbildung zu erhöhen.
    Notes: Abstract A nonporous ceramic implant in rat femora was evaluated as to its ability to bond to bone. A number of morphologic techniques were utilized to examine the interfacial relationship of the implant to new bone. Transmission electron microscopy and fluorescence microscopy after tetracycline labelling were the most successful techniques. An intimate relationship between mineralized bone and the ceramic was demonstrated by transmission electron microscopy. The appearance of tetracycline labelling at the ceramic interface indicates that the implant may have capacity to enhance bone formation.
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    Calcified tissue international 33 (1981), S. 143-151 
    ISSN: 1432-0827
    Keywords: Calcium transport ; Cytochalasin B ; Dihydrocytochalasin B ; Colchicine ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary In vivo calcium absorption was studied in normal and rachitic chicks. Cytochalasin B (CB) at a concentration of 25 µg/ml added to the medium inside the duodenal lumen inhibited calcium absorption (20 min) from 82.5±1.9% of calcium absorbed in the controls to 59.2±3% in normal and from 70.0±2.3% to 47.0±2.1% in rachitic chicks. In vitro studies by everted ileal sacs of young rabbits also showed an inhibition of active transport of calcium due to CB. Whereas in the controls the ratio of45Ca concentrations in serosal and mucosal media (60 min) was 7.2±0.32, the ratios were 5.24±0.52; 4.40±0.36; 3.40±0.42; 5.77±0.52; 1.38±0.08; and 1.06±0.02 in the presence of CB at concentrations of 5, 10 and 25 µg/ml; colchicine 10−4M, Na citrate 0.02M, and heat-devitalized conditions, respectively.45Ca concentration in the mucosal scrapings was also affected. It showed an increase from controls (15,101±404 cpm/mg) and correlated with CB concentration: 17,378±489, 19,015±1000, and 20,201±362 at 5, 10, and 25 µg/ml, respectively. Dihydrocytochalasin B also inhibited active calcium transport and caused an increase in45Ca concentration in the mucosal scrapings. Correlated electron microscopic studies showed certain changes in the brush border, especially in some actin microfilaments in the terminal web region. It seems that these morphological alterations may be related to transcytoplasmic movement of calcium.
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  • 35
    ISSN: 1432-0827
    Keywords: Connective Tissue ; Vitamin D deficiency ; Hydroxyproline excretion ; Glycosaminoglycan excretion ; Rat
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    Notes: Summary The urinary excretion of two connective tissue metabolites was studied in both control and vitamin D deficient rats. Hydroxyproline (HyPRO) excretion was determined after 2, 13 and 22 months (experiment I). It decreased with aging in animals receiving the control diet. On the contrary, this excretion increased as a function of age in vitamin D deficient animals. At the age of 22 months, HyPRO excretion was respectively 31 and 1708 µg a day in control and deficient animals. HyPRO and glycosaminoglycans (GAG) excretion was measured on a group of both control and vitamin D deficient rats at the age of 21 months (experiment II). These results confirm the high excretion of HyPRO in deficient animals. On the contrary, the GAG excretion was higher in control animals than in deficient ones, the mean excretion being respectively 412 and 234 µg a day.
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    Calcified tissue international 33 (1981), S. 223-231 
    ISSN: 1432-0827
    Keywords: Protein deficiency ; Bone formation ; Skeletal development ; Rat
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    Notes: Summary Protein deficiency was produced by freely feeding young rats a 1% lactalbumin diet for 12 weeks in order to study the effects of protein-calorie malnutrition on skeletal development. During the experiment the food and caloric intake and weight of the experimental animals decreased, while those parameters of the control animals progressively increased. However, when gross caloric intake was expressed as a function of the metabolic size of the animal, the caloric consumption was similar for both groups of animals. The protein-deficient animals exhibited micro-radiographic and histological features of an abnormal pattern of endochondral bone formation. Appositional bone growth, as determined by the daily appositional rate and the percentage of endosteal surfaces undergoing active bone formation, was significantly decreased in these animals, as was the percentage of periosteal surfaces exhibiting resorption. Both chemical analyses of the whole bone and electron probe microanalysis in the specific area of actively calcifying bone revealed no significant differences between the mineral content of control and protein-deficient animals. This study distinguishes the effects of protein deficiency from that of combined protein-calorie deprivation and demonstrates that the abnormal skeletal development observed was the result of a decrease in the quantity of bone formed rather than an altered mineral content.
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    Calcified tissue international 14 (1974), S. 283-289 
    ISSN: 1432-0827
    Keywords: Sodium fluoride ; Osteoporosis ; Bone ; Strength ; Rat
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    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé L'utilisation thérapeutique de fluorure de sodium a été recommandée dans le traitement de diverses maladies osseuses ostéopéniques. Cet emploi est basé principalement sur les effects ostéosclérotiques bien connus du fluorure de sodium, mais on connait mal son effet sur la force de résistance osseuse. L'influence de diverses concentrations de fluorure de sodium sur la force de résistance osseuse de jeunes rats soumis à des régimes riches et pauvres en calcium a été étudiée. L'administration de fluorure de sodium augmente le diamètre osseux, indiquant une stimulation de la formation de l'os périosté, mais la force de résistance osseuse est réduite ou non modifiée par l'ingestion de fluor.
    Abstract: Zusammenfassung Die therapeutische Verwendung von Natriumfluorid ist für eine Anzahl von Knochenmangel-Krankheiten empfohlen worden. Die Empfehlungen basieren hauptsächlich auf den bekannten osteosklerotischen Wirkungen von Natriumfluorid; über dessen Effekt auf die Knochenstärke ist wenig bekannt. In dieser Arbeit wurde der Einfluß verschiedener Konzentrationen von Natriumfluorid auf die Knochenstärken von wachsenden Ratten mit hoher und niederer Calciumeinnahme untersucht. Die Verabreichung von Natriumfluorid erhöhte den Knochendurchmesser, was auf eine Stimulierung der Periostbildung hinwies, die Knochenstärke wurde jedoch durch Fluorideinnahme herabgesetzt oder nicht beeinflußt.
    Notes: Abstract The therapeutic use of sodium fluoride has been recommended in a variety of osteopenic bone diseases. The recommendations are based mainly on the known osteosclerotic effects of sodium fluoride and little information is available as to its effect on bone strength. The influence of various concentrations of sodium fluoride on bone strength in growing rats on high and low calcium diets was studied. The administration of sodium fluoride increased bone diameter, indicating stimulation of periosteal bone formation, but bone strength was reduced or not affected by fluoride ingestion.
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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 26 (1978), S. 181-190 
    ISSN: 1432-0827
    Keywords: Cellular calcium ; Electron microscopy ; Osteoblasts ; Chondrocytes ; Mineralization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The calcium distribution in cartilage and bone cells during beginning ossification of fetal mouse long bones was studied after fixation with 2% K-pyroantimonate in 1% osmium. In the developing periosteum, the future osteoblasts showed a sparse cation-antimonate precipitate over the cytoplasm. In young osteoblasts the precipitate was accumulated on the mitochondrial membranes and the plasmalemma. Both organelles were sharply outlined by precipitate in the mature osteoblasts at the onset of mineralization. X-Ray microprobe analysis of these organelles demonstrated the presence of both Sb and Ca. In the extracellular compartment, a collagen-associated precipitate with 50 to 60 nm periodicity appeared during osteoblastic differentiation. During the initial phase of matrix mineralization, a random gross precipitate appeared in the matrix and seemed to be accumulated by osmiophilic matrix vesicles while the collagen fibrils lost their precipitate. Subsequently, during the confluent phase of matrix mineralization, the precipitate rapidly disappeared from the cells, leaving them devoid of precipitate once they were surrounded by mineralized matrix. Similar changes were found in the chondrocytes of the growth plate, but cartilage collagen, unlike osteoid collagen, did not bind precipitate. The results indicate that both osteoblasts and calcifying cartilage cells bind calcium prior to matrix mineralization. Bone collagen has strong pyroantimonate binding capacity, but it is not directly involved with initial stages of matrix mineralization, which starts in close association with matrix vesicles.
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    Calcified tissue international 25 (1978), S. 75-83 
    ISSN: 1432-0827
    Keywords: Rat ; Fluorosis ; Enamel ; Scanning electron microscopy ; Low temperature incineration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Sixteen 58-day-old male rats of Wistar strain, with a mean body weight of 179 g, were divided into two equal groups. Each group of eight animals was maintained for 70 days on drinking water, ad lib., containing no fluorine (control group) and 100 ppm of fluorine (experimental group). All specimens examined were obtained from the incisal portions of the incisors. The following types of enamel specimens were prepared for scanning electron microscopy: (1) acid-etched specimens; (2) acid-etched specimens followed by low temperature microincineration; and (3) fractured specimens. The enamel formed during high fluoride exposure showed marked hypocalcification, that is, the crystallite density in the prism core and interprismatic region was lower than that of control animals. The organic substances appeared to increase in these regions. These changes were prominent in the outer and middle enamel layers. Such changes following fluoride administration appear to indicate an inhibition of enamel maturation, that is, an inhibition of the mineral deposition and/or an inhibition of organic matrix withdrawal.
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  • 41
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    Calcified tissue international 23 (1977), S. 161-170 
    ISSN: 1432-0827
    Keywords: Bone ; Growth ; Rat ; Remodeling ; Tetracyclines
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Cortical bone remodeling along the femur diaphysis was determined in normal female rats (Sprague-Dawley) with the tetracycline technique. Three segments on the cortical bone circumference (the anterolateral, the medial, and the posterior) were found to be most suitable for the study of the remodeling process. Oxytetracycline was administered at age 60 and 75 days, and groups of animals were killed at age 75, 85, 95, and 105 days. The accumulated endosteal growth during age 60 to 75 days in the anterolateral segment was found to increase uniformly in the distal direction along the femur diaphysis. A method is described where this accumulated endosteal growth is used. This method eliminates the use of calipers to determine the section level and makes it possible to study comparable sections even after varying periods of time. The proximal part of the diaphysis showed periosteal apposition in all three segments. The periosteal apposition turned into resorption in the distal part of the diaphysis in the anterolateral and medial segments, whereas the periosteal appsition increased in the posterior segment. The endosteal growth increased in the distal direction in the anterolateral and medial segments. Irregular OTC bands made measurements of endosteal remodeling in the posterior segment impossible. The cortical width decreased in the distal direction along the femoral shaft. Comparison between the different age groups is described and also the relation between the accumulated endosteal growth and the diameter of the medullary cavity.
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  • 42
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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
    Source: Springer Online Journal Archives 1860-2000
    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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  • 43
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    Calcified tissue international 27 (1979), S. 161-164 
    ISSN: 1432-0827
    Keywords: Osteoporosis ; Ovary ; Rat ; Vitamin D
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Fifty adult female rats were used to study the effect of 1-alpha-hydroxyvitamin D3 (1α-OH-D3)on bone after oophorectomy. The experimental period was 6 months. At the end of the experiment the femurs and the tibias were investigated for bone mass and composition. Significant signs of osteopenia occurred as a result of oophorectomy. The treatment with 1α-OH-D3 induced only minor changes in blood chemistry but increased bone mass significantly. The findings support the view that 1α-OH-D3 may be a valuable tool in the treatment of osteoporosis resulting from ovarian insufficiency.
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  • 44
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    Calcified tissue international 34 (1982), S. 211-213 
    ISSN: 1432-0827
    Keywords: Rat ; incisor ; ameloblasts ; enamel ; 45Ca autoradiography
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Rats were injected with45Ca and horseradish peroxidase to determine the patterns of45Ca incorporation into incisor enamel and the morphological types of the overlying maturation ameloblasts.45Ca autoradiography showed no differences in the patterns of incorporation into enamel between routinely embedded and freeze-dried specimens. Enamel overlaid by ruffle-ended ameloblasts was much more heavily labeled while that overlaid by smooth-ended ameloblasts showed only moderate labeling. The observations lend further support to the hypothesis that the ruffle-ended cells are very active in mineralizing enamel and that the smooth-ended cells are in a passive, restorative phase.
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  • 45
    ISSN: 1432-0827
    Keywords: Bone alkaline phosphatase ; Rat ; Vitamin D ; Sorbitol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary In the femoral extremities of the adult rat containing the metaphysis, the epiphyseal cartilage, and the epiphysis, four alkaline phosphatase (AP) forms were distinguished on polyacrylamide gel electrophoresis. Two soluble forms were present in the 160,000 g supernatant: one of Mr 165 kDa and another of Mr 110–115 kDa, which exhibited a strong catalytical activity. Moreover, from the pellet, three membrane-bound forms of Mr 130, 110–115, and 100 kDa could be extacted with sodium deoxycholate. When denaturated AP was visualized by postelectrophoretic autoradiography of the phosphorylated intermediates, subunits always appeared as three monomers of Mr 75–80, 60–70, and 50–60 kDa. As four native forms but only three types of subunits were found to be present in the femur, it seems that, apart from homodimers, some heterodimers could also occur. Three types of diets were administered to three groups of rats for 5 weeks. Two are known to disturb bone mineralization: (1) a vitamin D3-deficient diet, and (2) the same as (1) but enriched with 12% sorbitol. The third was a normal diet containing vitamin D3. Concerning the effects on AP of dietary sorbitol and the vitamin D3-deficient diet, it was found that rats receiving the diet supplemented with sorbitol showed a substantial rise in the activity of the Mr 165 kDa form with the concomitant appearance of a new monomer of Mr 100 kDa. In contrast, rats fed the vitamin D3-deficient diet always displayed an increase in enzyme activity, principally of the Mr 100 and 110 kDa forms. In conclusion, the femur extracts of normal rats contained different forms of AP: either soluble 110–115 and 165 kDa forms or membrane-bound 130, 110–115, and 100 kDa forms. The administration of sorbitol-enriched diet induced a marked increase of the 165 kDa form whereas the administration of vitamin D3-deficient diet increased the 100 and 110 kDa forms.
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  • 46
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    Calcified tissue international 51 (1992), S. 137-142 
    ISSN: 1432-0827
    Keywords: Swimming exercise ; Bone histomorphometry ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary To specify the exercise-induced changes on different skeletal sites, the effect of a 5-week endurance swin training was studied in rats. Eighteen Lyon strain (Sprague-Dawley) 5-week old female rats were divided into nine sedentary and nine swimming rats. Each swim training session was increased by 15 minutes from 2–6 hours per day. A histomorphometric study was performed at the primary and secondary spongiosa of the distal femur and at the secondary spongiosa of lumbar and thoracic vertebral bodies. After training, bone loss was observed in the secondary spongiosa of lumbar vertebral bodies (24.7%) and in the primary spongiosa of distal femur (15.2%). A tendency to bone loss was also detected in the secondary spongiosa of distal femur (10.8%), whereas no change was detected in thoracic vertebral bodies. In secondary spongiosa, bone loss was accompanied with a thinning of trabeculae. Total eroded surfaces and osteoid surfaces were significantly decreased in the three studied skeletal sites, suggesting a decreased bone turnover. The decreased thickness of osteoid seams in both lumbar vertebrae and distal femur could mean that the osteoblastic activity has also been altered at the cell level, leading to thinning of trabeculae. Five-week swim training with such duration and intensity of exercise appears unable to increase bone volume in rats and, therefore, causes adverse effects. The three studied bones seemed to adapt differently to experimental conditions. The lack of ground reaction forces induced by water immersion might have contributed to the observed bone loss. “Normal” gravity would be an important cofactor in the osteogenic effects of exercise.
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  • 47
    ISSN: 1432-0827
    Keywords: Bone ; Nandrolone decanoate ; Ovariectomy ; Bone mechanics ; IGF-I ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Nandrolone decanoate (ND) is an anabolic steroid with a positive effect on bone mass in osteoporotic patients. The mechanism of action, (i.e., reduction of bone resorption and/or Stimulation of bone formation), the ultimate effect on mechanical properties, and the most effective dosage are not yet clear. To address these issues, dose-related effects of the long-term effect of ND on Serum and bone biochemistry, bone mineral content, and bone mechanical properties in ovariectomized (OVX) rats (12 weeks old at the Start of the experiment) were Studied for 6 months. The results were compared with those obtained in agematched, intact, and OVX rats. OVX caused in the femur a significant increase in net periosteal bone formation and net endosteal bone resorption of bone collagen content and torsional strength, and of Serum alkaline phosphatase, osteocalcin, and insulin-like growth factor-I (IGF-I) levels, whereas cortical bone density and calcium/creatinine and phosphorus/creatinine in 24-hour urine were Significantly reduced. Treatment of OVX rats with 1 mg ND/14 days resulted in a Significant increase in periosteal bone formation, femur length, cortical and trabecular bone mineral content and density, torsion stiffness and Strength, and bone IGF-I content, and a decrease in Serum osteocalcin, urinary calcium/creatinine levels, and bone collagen content compared with OVX controls. The higher ND dosage of 2.5 mg/14 days did not improve the results. ND treatment did not reverse all changes induced by OVX to the level of the intact controls. These results indicate that ND acts as an antiresorptive drug and as a bone formation Stimulating drug. Furthermore, the increased bone mass and bone mineral density is associated with improved bone Strength and stiffness and the presence of an increased amount of IGF-I. IGF-I is a growth factor considered to play a role in the maintenance of normal skeletal balance by a paracrine or autocrine mechanism.
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  • 48
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    Calcified tissue international 54 (1994), S. 150-154 
    ISSN: 1432-0827
    Keywords: Vitamin D analog ; KH1060 ; Kidney transplantation ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract KH1060 is a new 20-epi-vitamin D3 analog, which has exerted a considerable immunosuppressive potency in vitro. We have tested in vivo the effect of KH1060 on the suppression of renal allograft rejection in the rat. Allogenic kidney transplantation from DA donor rats to Lewis recipient rats treated intraperitoneally with KH1060 in doses from 0.2 to 6 μg/kg/day, or saline (placebo group), or CyA 10 mg/kg/day for 10 days (positive control group), was performed. Median graft survival time in KH1060-treated groups was 7–9 days, in the placebo group 6 days, whereas CyA led to long-term graft survival, 34 days in 50% of rats and 〉100 days in 50% of rats. In vivo, KH1060 failed to prolong renal allograft survival considerably, and led to development of hypercalcemia. Our results stress the existence of a large discrepancy between the in vitro and in vivo immunoregulatory effects of this vitamin D analog.
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  • 49
    ISSN: 1432-0827
    Keywords: Loading ; Strain ; Modeling ; Rat ; Ulna
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    Topics: Biology , Medicine , Physics
    Notes: Abstract Adaptive changes in bone modeling in response to noninvasive, cyclic axial loading of the rat ulna were compared with those using 4-point bending of the tibia. Twenty cycles daily of 4-point bending for 10 days were applied to rat tibiae through loading points 23 and 11 mm apart. Control bones received nonbending loads through loading points 11 mm apart. As woven bone was produced in both situations, any strain-related response was confounded by the response to direct periosteal pressure. Four-point bending is not, therefore, an ideal mode of loading for the investigation of strain-related adaptive modeling. The ulna's adaptive response to daily axial loading over 9 days was investigated in 30 rats. Groups 1–3 were loaded for 1200 cycles: Group 1 at 10 Hz and 20 N, Group 2 at 10 Hz and 15 N, and Group 3 at 20 Hz and 15 N. Groups 4 and 5 received 12,000 cycles of 20 N and 15 N at 10 Hz. Groups 1 and 4 showed a similar amount of new bone formation. Group 4 showed the same pattern of response but in reduced amount. The responses in Groups 2 and 3 were either small or absent. Strains were measured with single-element, miniature strain gauges bonded around the circumference of dissected bones. The 20 N loading induced peak strains of 3500–4500 μstrain. The width of the periosteal new bone response was proportional to the longitudinal strain at each point around the bone's circumference. It appears that when a bone is loaded in a normal strain distribution, an osteogenic response occurs when peak physiological strains are exceeded. In this situation the amount of new bone formed at each location is proportional to the local surface strain. Cycle numbers between 1200 and 12,000, and cycle frequencies between 10 and 20 Hz have no effect on the bone's adaptive response.
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  • 50
    ISSN: 1432-0827
    Keywords: Fluoride ; Bone ; Defluoridation ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract The aim of this work was to explore the reduction of fluoride concentrations in the skeleton after stopping experimental fluoride administration. Fluoride was administered to the rats at varying doses (0, 50, 100 ppm in drinking water) and for different lengths of time (4, 13, 25 weeks). A series of fluoride concentrations across the full thickness of humerus, parietal bone, and vertebra arch in rats were measured by means of an abrasive micro-sampling technique. The distribution profiles of fluoride from periosteal to endosteal surfaces, which were apparently related to the histological structure of these bones, were U shaped in the humerus, V shaped in the parietal bone, and W shaped in the vertebra arch. The average fluoride concentrations in the bones increased significantly with each increasing dose and length of fluoride administration. The relative increments were similar between the different regions or the different bones. After stopping fluoride administration, on the other hand, the relative reduction of the average fluoride concentrations in the bones were 30–100%. They were greatly related to the length after stopping fluoride administration and the dose and length of fluoride administration, but also dependent upon the type of bone and the region examined.
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  • 51
    ISSN: 1432-0827
    Keywords: Bone sialoprotein ; osteoblast ; Bone matrix ; Electron microscopy ; Immunolocalization ; noncollagenous protein
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    Notes: Abstract Bone sialoprotein was immunolocalized at the EM level in thin Lowicryl K4M sections of rat bone. Because of the unconventional EM morphology of the bone matrix seen in thin demineralized acrylate sections, the pattern of immunolabeling was compared with detailed structural images of demineralized bone obtained using an en bloc treatment of tissue samples with the cationic electron ‘dye’, Malachite Green (MG), which provides stabilization and retention of anionic material throughout specimen processing. A system of structures corresponding to the sites of bone sialoprotein (BSP) immunoreactivity, as seen in Lowicryl K4M thin sections, could be readily identified in the MG-treated, expoxy thin sections. This system includes the cement lines, and aggregates of similar material within mineralized bone and mineralizing osteoid. The virtual identity of BSP distribution with the arrangement of the MG-visualized material indicates that a BSP-enriched, noncollagenous phase can be demonstrated using different, unrelated tissue preparation and imaging protocols for EM. Besides improving our understanding of the distribution of bone sialoprotein in bone, these data assign a previously unrecognized structural dimension to noncollagenous material in the bone matrix.
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  • 52
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    Calcified tissue international 8 (1971), S. 287-303 
    ISSN: 1432-0827
    Keywords: Calcification ; Bone ; Matrix ; Apatite ; Nucleation ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Description / Table of Contents: Résumé Du collagène d'os compact de mouton est préparé par décalcification dans I'EDTA et à partir de tendons de queux de rats, par extraction dans l'acide acétique et reconstitution dans NaCl. Le dépôt d'apatite dans le collagène osseux de mouton dans une solution de calcification métastable est étudié chimiquement et par microscopie électronique. Le collagène osseux est un bon catalyseur de nucléation pour le dépôt minéral, alors que le collagène de tendons de rat ne l'est pas. Le dépôt minéral du collagène osseux se produit en deux phases cinétiques séparées, une phase rapide de nucléation et une croissance cristalline, donnant naissance à de petits ilots calcifiés et une seconde phase lente de croissance dans des régions ne comportant pas de zones catalytiques. La seconde phase de dépôt minéral paraît être le résultat d'une diffusion inhibée d'ions à travers les fibrilles collagènes alignées, laissant de larges régions de collagène sans minéral, bien que le tampon reste hautement sursaturé. La microscopie électronique permet de penser que les zones de catalyse pourraient avoir un rapport avec la périodicité de 640 Å de collagène, mais l'importance d'un matériel noncollagènique, lié au collagène, n'est pas à exclure. L'activité catalytique faible du collagène reconstitué n'est pas liée à la présence d'inhibiteurs faiblement liés, bien que des inhibiteurs puissent être intimement liés à ce type de collagène, qui pourrait être absent du collagène osseux. La différence d'activité catalytique pourrait intervenir dans la calcification physiologique. Une hypothèse plus générale pour la nucléation de la phase minérale dans les systémes biologiques est nécessaire.
    Abstract: Zusammenfassung Kollagen wurde aus kompaktem Schafsknochen mittels EDTA-Entkalkung und aus Rattenschwanzsehnen durch Essigsäureextraktion und Rekonstitution mit NaCl gewonnen. Die Apatitablagerung aus einer metastabilen Verkalkungslösung auf Schafsknochenkollagen wurde chemisch und im Elektronenmikroskop untersucht. Es zeigte sich, daß das Knochenkollagen ein guter Nukleationskatalysator für die Mineralablagerung ist, was beim Rattenschwanzkollagen nicht zutraf. Im Knochenkollagen erfolgte die Mineralablagerung in zwei getrennten kinetischen Phasen: einer raschen Phase der Nukleation und des Kristallwachstums, welche kleine verkalkte Inseln entstehen läßt, und einer zweiten langsamen Phase, welcher das Wachstum in Bezierken, die keine katalytischaktiven Stellen einschließen, zuzuschreiben ist. Diese zweite Phase der Mineralablagerung wird als Resultat einer verminderten Ionendiffusion durch die enganeinanderliegenden Kollagenfibrillen angesehen, wodurch weite Kollagenbereiche ohne Mineral bleiben, obwohl der Puffer stark übersättigt ist. Elektronenmikrographien ließen vermuten, daß die katalytischaktiven Stellen in einem gewissen Verhältnis zur 640 Å-Periodizität des Kollagens stehen; es konnte jedoch nicht ausgeschlossen werden, daß nicht-kollagenhaltiges Material, welches an Kollagen gebunden ist, ebenfalls eine Rolle spielt. Die schlechte katalytische Aktivität des rekonstituierten Kollagens konnte nicht auf die Anwesenheit von schwachgebundenen Hemmstoffen zurückgeführt werden, obwohl Inhibitoren stark an dieses Kollagen gebunden sein könnten, die jedoch im Knochenkollagen nicht vorhanden sind. Die Unterschiede in der katalytischen Aktivität können mit der physiologischen Verkalkung in Beziehung stehen. Eine allgemeinere Hypothese für die Nukleation einer Mineralphase in biologischen Systemen wäre erforderlich.
    Notes: Abstract Collagen was prepared from compact sheep bone by decalcification with EDTA and from rat tail tendons by acetic acid extraction and reconstitution with NaCl. The deposition of apatite in sheep bone collagen in a metastable calcification solution was studied chemically and by electron microscopy. The bone collagen was shown to be a good nucleation catalyst for mineral deposition, while rat tail collagen was a poor catalyst. Mineral deposition in bone collagen occured in two separate kinetic phases, a rapid phase of nucleation and crystal growth, giving rise to small calcified islands, and a second slow phase, ascribed to growth in regions not involving the catalytic sites. This second phase of mineral deposition is considered to be the result of impaired ion diffusion through the closely-aligned collagen fibrils, thus leaving large areas of the collagen free of mineral even though the buffer remains highly supersaturated. Electron micrographs suggested that the catalytic sites might be in some relationship to the 640 Å periodicity of collagen, but a role for non-collagenous material bound to the collagen has not been excluded. The poor catalytic activity of reconstituted collagen was not due to the presence of loosely-bound inhibitors, although inhibitors could be strongly bound to this type of collagen and be absent from bone collagen. The differences in catalytic activity may have a bearing on physiological calcification. A more general hypothesis for nucleation of a mineral phase in biological systems is required.
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  • 53
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    Keywords: Alcohol ; Electron microscopy ; Growth plate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary We have previously demonstrated that ethanol has a direct toxic effect on the rat skeleton characterized by decreased trabecular bone volume. In the present study, we examined the ultrastructure of the distal radial epiphyseal growth plates in these same animals. Eight weeks of ethanol administration to 12 male rats results in serum alcohol levels of 140 mg/dl but did not alter the width or light microscopic appearance of the radial growth plate. Quantitative electron microscopy failed to demonstrate morphologic evidence of toxicity in the skeletal cells. We conclude that although ethanol appears to have a direct effect on rat bone characterized by enhanced resorption, toxicity is not attended by ultrastructural changes in the skeletal cells.
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  • 54
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    Calcified tissue international 33 (1981), S. 529-540 
    ISSN: 1432-0827
    Keywords: Bone cells ; Electron microscopy ; PTH ; PGE1
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    Topics: Biology , Medicine , Physics
    Notes: Summary Four different cell populations—designated PF, OB, OC, and PC—were isolated from calvaria of 18-day-old chick embryos for analysis of the effects of hormones on bone tissue. The cell populations were studied with histological and biochemical methods. Apart from the well-known cell types present in calvaria, a new cell type was found in the noncalcified organic matrix between the osteoblastic layer and the calcified matrix. These cells were provisionally called osteocytic osteoblasts. They represent the “transition state” between osteoblasts and osteocytes. On the basis of histological studies with light microscopy (LM), transmission electron microscopy (TEM) and scanning electron microscopy (SEM), the PF population was considered to originate primarily from the periosteal fibroblasts, the OB population from the osteoblasts and osteocytic osteoblasts. The population of cells still present in calvaria after removal of periosteal fibroblasts and osteoblasts was called the OC population. This cell population was very much enriched with osteocytes. The fourth isolated population (PC) was a mixed population of fibroblasts, osteoblasts, and preosteoblasts. On exposure to parathyroid hormone (PTH), all four cell populations showed increased lactate production, but only the OB and OC populations displayed increased cAMP production. Prostaglandin E1 (PGE1) stimulated cAMP production in both OB and PF cells. From the results of this study it was concluded that PTH receptors are present on all of the cell types studied, but that occupancy of the receptor induces adenylate cyclase stimulation only in osteocytes and fully differentiated osteoblasts.
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    Calcified tissue international 15 (1974), S. 213-220 
    ISSN: 1432-0827
    Keywords: Acid phosphatase ; Electron microscopy ; Shell Regeneration
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    Topics: Biology , Medicine , Physics
    Notes: Abstract Acid phosphatase activity was mainly localized in the lysosomes in all the regions of the outer epithelium. The transitional portion of the outer epithelium showed more intense activity than the other regions. During shell regeneration the activity of this portion decreased to a minimum level at 12 hours and was restored to normal at 72 hours. The other regions showed no change of activity during shell regeneration. It is postulated that the acid phosphatase in the transitional protion is responsible for conferring calcifiability to the organic matrix of the shell.
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  • 56
    ISSN: 1432-0827
    Keywords: Cortisol ; Cortical bone ; Bone biomechanics ; Rat ; Femur
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Doses of 8, 16 (low), 32, 48, 64 (medium), and 150 (high) mg/kg/day of cortisol were administered to groups of 8 growing rats each during 16 days, and their femurs were then submitted to 3-point bending tests at low strain rate. Low doses had no effect. Medium doses, previously shown to improve calcium (Ca) balance and weight gain in the species, augmented diaphyseal elastic and ultimate strength, stiffness, and plastic-to-elastic deformation ratio with respect to untreated controls. This effect was achieved either by enhancing bone mass (volume, sectional moment of inertia, wall/lumen ratio) without changes in material quality parameters (32 mg/kg/day) or, conversely, by increasing bone tissue mechanical properties (stress, modulus of elasticity) not affecting bone geometry (48 and 64 mg/kg/day). The highest dose, known to depress Ca balance and weight gain, impaired diaphyseal mechanical performance in controls by substantially reducing bone mass without major variation in bone material properties, that is, developing a true osteopenic state in mechanical terms. The energy elastically absorbed per unit volume (proportional to the risk of comminute fractures) was greater with the highest dose because of enhanced deformability and diminished bone mass. The biphasic dose-response curves obtained, grossly parallel to those previously demonstrated for metabolic actions of cortisol in the same species, showed that biomechanical repercussion of this treatment on bone depends on different, dose-dependent effects which vary independently in temporal course, intensity, and sign.
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  • 57
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    Calcified tissue international 51 (1992), S. 30-34 
    ISSN: 1432-0827
    Keywords: PTHRP ; Anabolic ; Rat ; Bone ; Potency
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Parathyroid hormone-related protein (PTHRP) has recently been purified from human tumors associated with the syndrome of humoral hypercalcemia of malignancy. The gene encoding PTHRP has been cloned, and based on predicted amino acid sequence, polypeptides comprising the first 36 [36Tyr(1–36) PTHRP amide] and 74 [(1–74)PTHRP] amino acids have been synthesized. Human (h) PTHRP (1–36) and (1–74) are potent bone-resorbing agents, and are catabolic for bone in vivo when given continuously at high doses. Bovine parathyroid hormone (bPTH) (1–34) is also catabolic for bone at high dose levels, but when given in low doses for weeks to months, it is anabolic. Although PTHRP possess several PTH-like properties in bone, hPTHRP (1–34) is reported to be only weakly anabolic in vivo. As polypeptide length influences PTHRP action, we evaluated hPTHRP(1–74) as an anabolic agent for bone in vivo. Twenty-four 4-week-old male Sprague-Dawley rats were given daily subcutaneous injections of hPTHRP(1–74) (1 and 2 nmol/100 g body weight, bw), bPTH(1–34) (4 nmol/100 g bw) or vehicle. Rats were sacrificed on day 12, and serum calcium, phosphorus, and 1,25 dihydroxyvitamin D and femoral bone dry weight, calcium content, and hydroxyproline content were measured. Serum calcium and phosphorus were equivalent in all groups. A significant increase in dry bone weight was observed in both PTHRP-treated groups compared with controls. PTHRP also caused a significant, dose-dependent increase in bone calcium and hydroxypro-line content. Results of these studies indicate that PTHRP (1–74) is anabolic for bone in vivo when administered at low-dosage levels for a prolonged period.
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  • 58
    ISSN: 1432-0827
    Keywords: Chondrocytes ; High-density suspension culture ; Electron microscopy ; Matrix vesicle ; Apatite formation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Chondrocyte cultures grown in centrifuge tubes with intermittent centrifugation differentiate into hypertrophic chondrocytes and form calcification. We examined chondrocytes cultured in this system electron microscopically. Rat growth-plate chondrocytes were seeded in a plastic centrifuge tube and cultured in the presence of Eagle's minimum essential medium supplemented with 10% fetal bovine serum and 50 μg of ascorbic acid per ml. Specimens were examined by using electron microscopy and selected-area electron-diffraction techniques. In the early stage of culture, a few chondrocytes were scattered and extracellular matrices were not observed. In the middle stage of the cultures, the chondrocytes resembled proliferative cells. Matrix vesicles appeared to be budding from the cell surfaces of chondrocytes and were observed sparsely in the extracellular matrices, which were well formed around the chondrocytes. Matrix vesicles increased substantially during the following cultures. In the mature stage of the cultures, crystal formation related to matrix vesicles was observed. In the 33-day cultures, several masses of calcified matrix were formed and it was confirmed to be apatite by selected-area electron diffraction analysis. The chondrocytes appeared hypertrophic during this same stage. The 56-day culture was similar to the 33-day culture. It was concluded that this culture system provides an extracellular-matrix mineralization which is produced by chondrocytes per se.
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    Calcified tissue international 55 (1994), S. 33-37 
    ISSN: 1432-0827
    Keywords: Osteoporosis ; Rat ; Orchidectomy ; Exercise ; Strength
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    Topics: Biology , Medicine , Physics
    Notes: Abstract The effect of exercise on castration-induced osteoporosis in 3-month-old male rats weighing 264±4 g at the beginning of the experiment was studied. A testosterone deficiency was induced by orchidectomy (ORC), and the exercise group ran 10 m/minute for 1 hour a day on a treadmill at 0% grade. There were seven groups of eight rats (n=56) randomized into a control group killed at time 0, and sham, ORC and ORC and exercise groups killed at 4 and 8 weeks. ORC reduced body weight gain (with analysis of variance (ANOVA) P〈0.001), and at 4 weeks the body weight was 343±14 g in ORC group and 301±4 g in the ORC and exercise group (P〈0.01). The increase in femoral length was slower in the ORC+exercise groups. The ash weight of the tibia did not decrease significantly after ORC or ORC+ exercise. ORC did not affect 45Ca incorporation, but exercise slightly increased it in the whole tibia 8 weeks after ORC (with ANOVA P=0.057). ORC had significantly lowered the trabecular bone volume in the secondary spongiosa of the distal femur at 4 and 8 weeks, and exercise did not prevent this. This is an opposite finding to our previous study with ovariectomized female rats [12]. ORC also significantly had reduced the osteoblast-lined trabecular bone surface and the number of osteoclasts by 8 weeks after the operation. Exercise increased the osteoblast-lined surface and the number of osteoclasts. The mechanical strength of the femoral neck also was reduced after ORC and this was not prevented by exercise either. In conclusion, ORC reduces bone growth and turnover which leads to osteopenia in growing rats. Moderate treadmill exercise does not reverse the ORC-induced loss of trabecular bone and the reduced mechanical strength of the femoral neck, although it has a positive effect on the osteoblast and osteoclast indices and on calcium incorporation into bone.
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  • 60
    ISSN: 1432-0827
    Keywords: Bone mechanics ; Bone composition ; Vitamin D3 ; Corticosteroid ; Rat
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    Topics: Biology , Medicine , Physics
    Notes: Abstract The effects of 1α-vitamin D3 were studied for 6 months in 2-month-old male and female rats on a moderately low calcium diet with or without low-dose prednisolone treatment. Both cortical bone mechanical and biochemical properties were examined. Femoral bone specimens were subjected to torsional loading tests. With age, bone strength and stiffness increased in both sexes, accompanied by an increased degree of mineralization (bone ash and calcium concentrations). During growth, strength and stiffness increased more in male than in female rats. When 1α-vitamin D3 (0.5 μg/kg/day) was given alone, bone mechanical competence improved significantly whereas insulin-like growth factor-I (IGF-I) and calcium concentrations in the bone matrix were significantly reduced. Treatment with low-dose prednisolone (0.5 mg/kg/day) alone did not influence bone mechanical properties compared with intact control rats (without prednisolone) although a significant reduction in calcium concentration and an increased phosphorus concentration were measured. A combined therapy with prednisolone and 1α-vitamin D3 significantly increased bone strength, toughness, and stiffness compared with control bones. Both mineralization degree (ash and calcium concentration) and IGF-I concentration were decreased. We conclude that (1) mechanical properties of rat cortical bones improve relatively more in males compared with agematched females during growth which is related to increased bone mass and size, (2) low-dose prednisolone treatment does not change mechanical properties in males, and altered them only nonsignificantly in females despite a change in mineralization degree in both sexes; (3) treatment with 1α-vitamin D3 results in a consistent increase in mechanical competence of the bone accompanied by a significant decrease in IGF-I concentration in the bone matrix.
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  • 61
    ISSN: 1432-0827
    Keywords: Cathepsin inhibitors ; Osteoclasts ; Resorption ; Electron microscopy
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    Topics: Biology , Medicine , Physics
    Notes: Abstract This study was designed to evaluate the effects of specific and potent cathepsin inhibitors on osteoclastic resorptive functions in vitro by means of a novel ultrastructural assay system. Mouse bone marrow cell-derived osteoclasts were suspended on dentine slices and cultured for 48 hours in the presence of either E-64 (a generalized cysteine proteinase inhibitor) or Z-Phe-Phe-CHN2 (a selective cathepsin L inhibitor). After the removal of cultured osteoclasts, co-cultured dentine slices were examined using electron microscopy: backscattered (BSEM), scanning (SEM), and atomic force (AFM). In morphometric analyses of BSEM images, there were no significant differences in the areas of demineralized dentine surfaces between control and inhibitor-treated groups, suggesting that cathepsin inhibitors had no effect on dentine demineralization by cultured osteoclasts. However, in SEM and AFM observations, both inhibitors remarkably reduced to the same extent, the formation of deep resorption lacunae on dentine slices that had resulted from degradation of matrix collagen. In addition, Z-Phe-Phe-CHN2 treatment produced deeper, ring-like grooves with little collagen exposure in shallow resorption lacunae. These results strongly suggest that (1) cathepsins released by osteoclasts are involved in the formation of deep resorption lacunae, and (2) cathepsin L plays a key role in bone resorption.
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    Cellular and molecular life sciences 40 (1984), S. 974-975 
    ISSN: 1420-9071
    Keywords: Rat ; prostaglandins ; gastric lesion ; intragastric distension model ; stress model ; indomethacin ; somatostatin preventive effect
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    Topics: Biology , Medicine
    Notes: Summary The inhibition of endogenous prostaglandin synthesis by indomethacin treatment blocks the somatostatin preventive effect on the gastric lesions induced in a stress model and has no preventive effect on an intragastric distension model.
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  • 63
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    Cellular and molecular life sciences 40 (1984), S. 1004-1006 
    ISSN: 1420-9071
    Keywords: Rat ; adrenocortical responsiveness ; ACTH ; plasma ; corticosterone ; plasma ; corticotropin releasing factor (CRF)
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    Topics: Biology , Medicine
    Notes: Summary In the course of studying the plasma adrenocorticotropic hormone (ACTH) and corticosterone responses to synthetic corticotropin releasing factor (CRF), we noted some disparity in the responses. A higher dose (20 μg compared with 5 μg per rat i.a.) produced an equal plasma ACTH but greater plasma corticosterone response in adult male rats. Thus, we examined the possibility that CRF increases adrenocortical responsiveness to ACTH. CRF significantly (p〈0.0005) increased the plasma corticosterone response to ACTH in rats pretreated with dexamethasone. Thus, synthetic CRF increases corticosterone secretion in rats not only by stimulating ACTH secretion, but also by increasing the adrenocortical responsiveness to ACTH.
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  • 64
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    Cellular and molecular life sciences 41 (1985), S. 273-275 
    ISSN: 1420-9071
    Keywords: Rat ; embryo ; embryo ; rat ; β-hydroxybutyrate ; diabetes ; maternal ; fetal malformation ; malformation ; fetal ; ketone bodies
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    Topics: Biology , Medicine
    Notes: Summary In order to investigate further the relationship between maternal diabetes and fetal malformation, rat embryos were grown in vitro in the presence of β-hydroxybutyrate, one of the ketone bodies produced by diabetics. At 10 mM, β-hydroxybutyrate produced minor abnormalities and at 20 mM it produced major abnormalities in rat embryos.
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  • 65
    ISSN: 1420-9071
    Keywords: Rat ; cerebrospinal fluid, human ; analgesia ; naloxone ; pain indifference, congenital ; opiates, endogenous
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    Notes: Summary CSF from a patient with congential indifference to pain was found to produce analgesia in the rat following intracerebroventricular injections. The analgesic effect was attenuated by pretreatment with naloxone suggesting the involvement of hyperactive endogenous opiate mechanisms in this patient.
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  • 66
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    Cellular and molecular life sciences 40 (1984), S. 1368-1369 
    ISSN: 1420-9071
    Keywords: Rat ; kidney ; hypertensive ; prostaglandin dehydrogenase ; hexokinase
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    Topics: Biology , Medicine
    Notes: Summary 15-Hydroxyprostaglandin dehydrogenase (PGDH) surged in hypertensive (SHR) and normotensive (WKY) rat kidney at 8 days of age, is greatest in SHR. Hexokinase fell in SHR at 17 days of age, but thereafter was similar to WKY. This suggests multisystem enzymatic abnormalities in SHR kidney during development of hypertension.
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  • 67
    ISSN: 1420-9071
    Keywords: Rat ; alloxan diabetes ; intestinal villus ; alkaline phosphatase ; sucrase ; epithelial cells ; villus-crypt axis
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    Notes: Summary The sucrase activity in enterocytes isolated from the villus crypt axis was found to increase in all regions of the villus from day 2 after induction of diabetes, and the increase continued until day 4. In contrast, alkaline phosphatase activity increased mainly in the apical one-third of the villus-crypt column, and the increase occurred abruptly on day 4 with increase in food intake.
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  • 68
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    Cellular and molecular life sciences 42 (1986), S. 169-171 
    ISSN: 1420-9071
    Keywords: Rat ; olfactory pathway selective lesions ; plasma corticosterone ; lactacidemia ; adrenal glands
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    Topics: Biology , Medicine
    Notes: Summary In comparison to control rats, basal plasma corticosterone level and lactacidemia significantly increased in rats submitted to a bilateral lesion of the lateral olfactory tract and/or the anterior branch of the anterior commissure. Only the anterior branch of the anterior commissure induced hyperglycemia; that of the lateral olfactory tract exerted an opposite effect.
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  • 69
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    Cellular and molecular life sciences 46 (1990), S. 457-458 
    ISSN: 1420-9071
    Keywords: Rat ; carnitine ; lipolysis ; exercise ; glycogen ; triglycerides ; muscle
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    Topics: Biology , Medicine
    Notes: Summary To test the effect of L-carnitine on glycogen sparing when fat oxidation is increased, 100 mg/kg/d were given to rats orally for 3 days, resulting in 1.8-fold higher muscle carnitine levels. Even when FFA were raised by heparin-stimulated lipolysis, the rate of glycogen degradation was not reduced during exercise.
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  • 70
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    Cellular and molecular life sciences 40 (1984), S. 1008-1010 
    ISSN: 1420-9071
    Keywords: Rat ; ethanol preference ; acetaldehyde self-administration
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    Topics: Biology , Medicine
    Notes: Summary Peripherally self-injected acetaldehyde in interaction with environmental and nutritional variables significantly enhances alcohol drinking in rats and suggests an involvement of acetaldehyde in voluntary alcohol intake.
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    Cellular and molecular life sciences 41 (1985), S. 265-266 
    ISSN: 1420-9071
    Keywords: Rat ; hydrochlorothiazide ; pain threshold ; antinociceptive activity ; analgesic activity ; morphine
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    Topics: Biology , Medicine
    Notes: Summary Hydrochlorothiazide, acutely injected in rats, has a weak analgesic activity per se and potentiates and prolongs the antinociceptive effect of morphine.
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  • 72
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    Trees 11 (1997), S. 378-387 
    ISSN: 0931-1890
    Keywords: Key words Pinus sylvestris (L.) ; Electron microscopy ; Heavy metals ; Multi-stress-symptoms ; SO2
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract  Injuries to needles of Scots pines (Pinus sylvestris L.) growing in nutrient-poor soils on the Kola Peninsula collected in April 1991 were studied on a gradient of increasing distances (10 – 115 km) from the Monchegorsk nickel smelter, Russia, which emits SO2, Ni and Cu. The condition of the mesophyll cells was quantified from needles of the two latest age classes using a light and an electron microscope. The damage to the ultrastructure consisted of multistress symptoms caused by excess sulphur, heavy metals, frost, acidic precipitation and ozone. Injuries were most commonly manifested in the form of dark, irregularly shaped chloroplasts with protrusions and light thylakoids and plastoglobuli. These symptoms gradually disappeared with increasing distance and decreasing deposition rate. Concentrations of sulphur, copper and nickel decreased towards more distant sites where normal levels of the latter two elements were reached. Sulphur concentrations remained above background throughout the distance gradient. In the closest plots to the smelter area, cell collapse under the stomata and epidermis related to acute SO2 and heavy metal effects was found, whereas further away symptoms were more diverse, pointing towards the effects of ozone, acidic deposition and thereby decreased frost tolerance. The additive multistress symptoms were clearly seen in the area up to 40 km from the smelter where needle Cu concentration was above 110 ppm, Ni concentration above 39 ppm and S concentration above 1343 ppm.
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  • 73
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    Journal of biomedical science 1 (1994), S. 201-203 
    ISSN: 1423-0127
    Keywords: Hypertension ; Eicosanoid ; Rat ; Genetics ; Kidney
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    Topics: Biology , Medicine
    Notes: Abstract The present paper reviews the evidence for a possible involvement of renal eicosanoids in the pathophysiology of high blood pressure in genetically hypertensive rats of the Lyon strain. Both in vivo and in vitro experiments suggest that an increased ability to synthesize the vasoconstrictor prostaglandin H2 and/or thromboxane A2 in renal vessels (1) acts as an autocrine amplifier of pressor agents and (2) may contribute to resetting the pressure natriuresis curve which is a prerequisite for the development and maintenance of hypertension.
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  • 74
    ISSN: 1432-041X
    Keywords: Synaptogenesis ; Electron microscopy ; Visual acuity ; Fish development
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    Topics: Biology
    Notes: Summary The morphogenetic differentiation of synapses of the optic tectum of the rainbow trout was investigated at different stages of development (from hatching to adult) and compared with the improvement in visual discrimination (minimum separable). (1) The main phase of synaptogenesis (increase in number of synapses, length of contact zone and number of vesicles) begins about one week after hatching and continues up to the age of one month, when the larvae start swimming freely. (2) Myelination begins 26 days after hatching and induces the end of the synaptogenesis period. (3) The visual discrimination (minimum separable) of trout larvae improves from 30 degrees of arc on the 10th day after hatching to 1 degree on day 30, then to about 14 to 18 min of arc in the adult. The results are discussed with special reference to previous biochemical investigations on changes in the ganglioside composition of the trout brain during comparable periods of development.
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    Development genes and evolution 191 (1982), S. 205-207 
    ISSN: 1432-041X
    Keywords: Chitin inhibition ; Nikkomycin ; Cuticle ; Electron microscopy ; Epilachna varivestis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The nucleoside antibiotic nikkomycin has proved to be an effective inhibitor of chitin synthesis in the Mexican bean beetleEpilachna varivestis. Ultrastructural investigations show defects in the procuticular area after nikkomycin application which suggest the complete absence of chitin. A cuticle like this is inflexible and too brittle to satisfy its normal function as an exoskeleton. The individuals are not able to free themselves from the exuvia and finally die. Therefore nikkomycin seems to be a potential insecticide with high specifity.
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    Development genes and evolution 206 (1997), S. 503-514 
    ISSN: 1432-041X
    Keywords: Key words Preimplantation mouse embryo ; Brefeldin-A ; Monensin ; Golgi ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The intracellular trafficking of integral membrane and secreted proteins is likely to be a key element involved in the morphogenesis and differentiation of the early mammalian embryo. In this study, we used transmission electron microscopy (TEM) to analyse the effects of brefeldin-A (BFA) and monensin, well known inhibitors of vesicular protein trafficking in somatic cells, on the structure of preimplantation mouse embryos. Both BFA and monensin distinctively altered the morphology of Golgi compartments in the blastomeres of treated morulae. BFA-treated morulae lacked recognizable Golgi complexes but possessed heterogeneous organelle clusters consisting of an abundance of smooth tubular and vesicular membrane compartments in addition to mitochondria, endosomes and lysosomes. Treatment of morulae with monensin was associated with swelling of Golgi compartments in addition to altering the morphology of mitochondria, lysosomes and the plasma membrane. BFA, and to a lesser extent monensin, inhibited cytokinesis as evidenced by the detection of binucleate blastomeres. In addition, BFA induced morulae to decompact. These latter effects have not been reported previously for these agents in mammalian somatic cell lines or other vertebrate or invertebrate embryos. These results provide the first demonstration of the structural effects of BFA and monensin on cells of the early mammalian embryo, some of which are consistent with the known actions of these agents on components of the vesicular protein trafficking system in mammalian somatic cells. This information serves as a foundation for the further use of these agents in studies of vesicular protein trafficking as an agent of preimplantation morphogenesis.
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  • 77
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    Calcified tissue international 25 (1978), S. 133-143 
    ISSN: 1432-0827
    Keywords: Osteon ; X-Ray diffraction ; Electron microscopy ; Calcification
    Source: Springer Online Journal Archives 1860-2000
    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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  • 78
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    Calcified tissue international 24 (1977), S. 191-197 
    ISSN: 1432-0827
    Keywords: Amelogenesis imperfecta ; Hypocalcification ; Hypoplasia ; Electron microscopy
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    Topics: Biology , Medicine , Physics
    Notes: Summary An ultrastructural study of teeth with amelogenesis imperfecta revelaed various aspects of microcavities in the enamel surface, which ranged from isolated imprints of ameloblasts corresponding to the mildest lesions at the end of amelogenesis, to pits caused by the death of 20 to 30 ameloblasts at the beginning of amelogenesis. Abnormalities in the shape of the prisms can be observed. Further, crystals are distributed randomly within a prism or at the junction of 2 contiguous prisms while intercrystalline spaces are widened, indicating in various places the lack of a preferred orientation of the crystals. In amelogenesis imperfecta, two different crystalline periods are found: 1 of about 250 Å, the other of about 500 Å and over. The fact that amorphous areas are found among the crystals of enamel may be related to different stages of crystallization. However, it was not possible to find any lattice defect.
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    Calcified tissue international 24 (1977), S. 239-242 
    ISSN: 1432-0827
    Keywords: Cementum ; Lysis ; Electron microscopy
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    Topics: Biology , Medicine , Physics
    Notes: Summary Examination of microradiographs from the deciduous teeth of pigs revealed large lacunae or radiolucent zones close to the cemento-dentinal junction. Electron microscopic studies of the ground sections showed areas or irregularly shaped zones devoid of mineral and filled with collagen fibers. In the wide unmineralized zones, spherical clusters of crystallites were noted. Several cementum lacunae bordered by a broad rim of unmineralized collagen fibers were noted and some lacunae also contained zones of a moderately electron dense material. This material did not yield a diffraction pattern, while the mineralized part of the cementum gave the diffraction pattern typical of hydroxyapatite.
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    Calcified tissue international 25 (1978), S. 217-222 
    ISSN: 1432-0827
    Keywords: Bone mineral ; Electron microscopy ; X-ray diffraction ; Dark field
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    Topics: Biology , Medicine , Physics
    Notes: Summary Electron microscopical observations of the size and shape of bone mineral crystallites have not been in complete agreement with X-ray diffraction findings. The two prevalent viewpoints consider bone mineral crystals to be either rod, or plate like in habit. There appears to be agreement that the smallest dimension of the crystals is about 5 nm, but there is discrepancy in the reported c-axial lengths. The method of dark field imaging is used to obtain a quantitative measurement of the c-axial length distribution in rabbit, ox and human bone: mean c-axial lengths 32.6 nm, 36.2 nm and 32.4 nm, respectively, show no significant difference at the 5% level to the mean c-axial length measured by X-ray line broadening. Both bright and dark field images strongly suggest that bone mineral has a plate like form. Reasons for past discrepancies are discussed.
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    Calcified tissue international 25 (1978), S. 45-51 
    ISSN: 1432-0827
    Keywords: CaCO3 ; Amino acids ; Sheaths ; Ligament ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The aragonite crystals in the molluscan bivalve hinge ligament are surrounded by an organic sheath which is distinct from the remainder of the ligament matrix. Methods have been developed to isolate these sheathed crystals from the ligaments ofSpisula solidissima andMercenaria mercenaria employing a papain digestion of the matrix protein. The sheathed crystals fromSpisula have a CaCO3/protein ratio of 11.1 and those fromMercenaria a ratio of 29.6. The sheathed crystals and the empty crystal sheaths have been examined by electron microscopy for structural integrity. The sheath proteins exhibit much smaller proportions of the amino acids glycine and methionine than the hinge ligaments. These are characteristic amino acids of high concentration in the hinge ligaments of both species. The concentrations of acidic and basic amino acids are increased about two fold in the sheaths over those of the ligaments. Otherwise there is little similarity in the amino acid composition of the sheaths in the two species. However, SDS electrophoresis shows the sheaths of both to contain a major protein component with a molecular weight of about 25,000. The sheath protein from theMercenaria ligament contains about 5% carbohydrate and that ofSpisula sheaths less than 1% carbohydrate.
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  • 82
    ISSN: 1432-0827
    Keywords: Rat ; Calvarium ; Electron microscopy ; Preosteoclasts ; Osteoclasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary This is a study of the fine structure of cells of the 20-day fetal rat calvarium. Special attention is given to identifying and characterizing preosteoclasts. These cells are relatively common and located largely, but not exclusively, at the endocranial bone surface. The preosteoclasts are characterized by abundant mitochondria, an incomplete perinuclear Golgi apparatus, and variable-shaped dense granules. The dense granules are unique in appearance in that they contain an internal dense matrix surrounded by a clear halo. Most granules are circular in shape but some are elongate or tubular in form. Granules with identical appearance are observed in osteoclasts. The preosteoclasts are mononucleate, or occasionally binucleate. It is suggested that because preosteoclasts are morphologically distinctive and relatively abundant, it should be feasible to separate these cells from a heterogeneous cell isolate.
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    Calcified tissue international 50 (1992), S. 524-526 
    ISSN: 1432-0827
    Keywords: Verapamil ; Bone ; Osteopenia ; Rat ; Female ; Intestinal calcium absorption
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Verapamil inhibits the intestinal absorption of calcium (Ca) and increases serum parathyroid hormone in rats. The effects of verapamil on bone tissue after long-term treatment is, however, not well described. Adult female and male Sprague-Dawley rats received verapamil in their drinking water at a dosage of 0.075 mg/ml (low dose) or 0.75 mg/ml (high dose) for 12 weeks; control rats received only drinking water. All rats were fed a diet containing 0.1% Ca and 0.5% P. In female rats, the amount of bone ash per volume was significantly reduced from 0.742 g/ml in controls to 0.713 g/ml after low-dose treatment of verapamil, and to 0.667 g/ml following high-dose treatment (P〈0.01). The tibial length was increased from 39.7 mm in controls to 40.3 mm or to 40.7 mm after low or high doses (P〈0.01). The tibial volume increased from 0.385 ml in controls to 0.397 ml after low doses and to 0.429 ml after high doses (P〈0.01). In contrast, in male rats the amount of bone ash per volume was significantly increased from 0.578 g/ml in controls to 0.580 g/ml after low doses and to 0.620 g/ml after high doses of verapamil (P〈0.01). The tibial bone volume in males as decreased from 0.633 ml in controls to 0.641 ml after low doses and to 0.583 ml after high doses (P〈0.05). The tibial length in the males was not changed by verapamil. The intestinal absorption of Ca was reduced in male rats from 5.28 in controls to 4.03 (serosa/mucosa) after low-dose treatment and to 2.46 after high-dose treatment with verapamil (P〈0.05). In female rats, the intestinal absorption of Ca did not change after verapamil treatment. Thus, chronic treatment with verapamil in female rats induced osteopenia whereas in male rats bone growth was inhibited.
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  • 84
    ISSN: 1432-0827
    Keywords: Rat ; Osteoblast-like culture ; Ovariectomy ; Estrogens ; Bone blood supply
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Ovariectomy in the rat induces a rapid osteopenia associated with an elevated bone turnover. One hundred and twenty-day-old rats were ovariectomized (OVX) or sham-operated (n=6–8 per group and per time period studied). 45Ca accretion rate and bone blood flow (microspheres trapping technique) in the femurs were determined at 28, 42, 84, and 119 days after ovariectomy. Both parameters were markedly increased by 84 days and subsided thereafter. At the 42nd day, when bone turnover was maximal, bone marrow and trabecular bone cultures were obtained from shamoperated and ovariectomized animals (n=10/group). Proliferation rate of bone marrow cells and trabecular osteoblast-like cells estimated by fibroblast colony-forming units (FCFU) efficiency and cell counting was markedly increased in primary and secondary cultures in ovariectomy. These data fitted well with the enhanced number of osteoblasts observed in situ in the long bone metaphyses of estrogen-depleted animals. As estrogens were shown in the literature to inhibit proliferation of the red cell line and of other hemopoietic lines, it is possible that estrogens, through a general mechanism, inhibit hemopoietic and stromal lines and also the proliferation of bone marrow-derived trabecular bone cells.
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  • 85
    ISSN: 1432-072X
    Keywords: d-Ribulose 1,5-diphosphate carboxylase ; Oxygenase activity ; Quaternary structure ; Electron microscopy ; Alcaligenes eutrophus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract d-Ribulose 1,5-diphosphate carboxylase has been purified from autotrophically grown cells of the facultative chemolithotrophic hydrogen bacteriumAlcaligenes eutrophus. The enzyme was homogeneous by the criteria of polyacrylamide gel electrophoresis. The molecular weight of the enzyme was 505000 determined by gel filtration and sucrose density gradient centrifugation, and a sedimentation coefficient of 18.2 S was obtained. It was demonstrated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis that the enzyme consists of two types of subunits of molecular weight 52000 and 13000. Electron microscopy on the intact and the partially dissociated enzyme lead to the construction of a model for the quaternary structure of the enzyme which is composed of 8 large and 8 small subunits. The most probable symmetry of the enzyme molecule is 4:2:2. Michaelis constant (K m ) values for ribulose 1,5-diphosphate, Mg2-, and CO2 were 0.59 mM, 0.33 mM, and 0.066 mM measured under air. Oxygen was a competitive inhibitor with respect to CO2 suggesting that the enzyme also exhibits an oxygenase activity. The oxygenolytic cleavage of ribulose 1,5-diphosphate was shown and a 1:1 stoichiometry between oxygen consumption and 3-phosphoglycerate formation observed.
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  • 86
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    Archives of microbiology 115 (1977), S. 185-198 
    ISSN: 1432-072X
    Keywords: Synechococcus lividus ; Cyanobacteria ; Carbon dioxide ; Electron microscopy ; Bleaching-regreening
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of carbon dioxide on pigment and membrane content in Synechococcus lividus was studied by depriving cells of CO2 and examining cell populations biochemically and by electron microscopy. After 120 h of CO2 deprivation, S. lividus lost all detectable chlorophyll a and C-phycocyanin. Such bleached cultures were “mustard yellow”, the result of approximately 1.8 times more carotenoid per cell than green control cultures. Although cells from beached cultures appeared morphologically identical to control green cells when examined by light microscopy, electron microscopic examination revealed them to be devoid of detectable thylakoid membrane. Thylakoid membrane could not be recovered by physical isolation or revealed by freeze etching of bleached S. lividus. In addition, inclusion bodies characteristically found in S. lividus were also absent. Reintroduction of CO2 into bleached cultures resulted in a rapid resynthesis of both chlorophyll a and C-phycocyanin. Electron microscopic examination of these regreening cultures revealed that thylakoid membrane was also rapidly resynthesized. Growth of regreened cultures did not occur until there was the synthesis of a full complement of chlorophyll a, C-phycocyanin, and thylakoid membrane. A time course study of the cytological events occurring during bleaching and regreening is presented.
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  • 87
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    Archives of microbiology 111 (1976), S. 175-183 
    ISSN: 1432-072X
    Keywords: Serratia marcescens ; (Phage tail) bacteriocin ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Description / Table of Contents: Zusammenfassung Die Genese eines phagenschwanzähnlichen Bacteriocins in Zellen des Gruppe A-bacteriocinogenen (bA+) Serratia marcescens-Stammer Nr. 16 wurde nach Mitomycin C (MC) Induktion elektronenoptisch untersucht. Dieses Bacteriocin (Gesamtlänge 117 nm) besteht aus einem hohlen Stift mit kontraktiler Scheide. Nach 60 min Induktion wurden in Dünnschnitten stäbchenförmige Bacteriocine identifiziert. Sie erscheinen in drei Aggregationsformen: 1. als hexagonale Einschlüsse, 2. als Bänder dicht nebeneinanderliegender Bacteriocine und 3. als Stapel von übereinanderliegenden Bacteriocinschichten, wenn nach 120 min Induktion ein Maximum von ca. 450 Bacteriocinen pro Zelle erreicht wird. Bacteriocine konnten nach der gleichen Induktionszeit von 60 min auch mit der “in situ lysis technique” nachgewiesen werden. Neben Bacteriocinen traten relativ selten und unregelmäßig auch Phagenköpfe auf. Die Stäbchenform teilungsfähiger Zellen blieb bis zum Auftreten von intracellulären Bacteriocinen erhalten. Ihre Umwandlung in geblähte, sphäroplastenähnliche Zellformen, die nach 120 min Induktion lysierten, war zeitlich korreliert mit Feinstrukturveränderungen der Zellwand.
    Notes: Abstract The biosynthesis of a phage tail-like Bacteriocin by cells of the group A-bacteriocinogenic (bA+ Serratia marcescens strain no. 16 after induction with mitomycin C (MC) was examined electronmicroscopically. This bacteriocin (total length 117 nm) consists of a hollow core and a contractile sheath. At 60 min following induction, rod-like bacteriocin-partieles were identifiable in ultrathin sections. The particles were found to comprise three morphologically different forms of aggregation: 1. hexagonal inclusions, 2. contiguous, bank-like particles, and 3. staples of superimposed layers of bacteriocin particles. At 120 min after induction bA+ cells revealed maximally 450 bacteriocin particles. Similarly, the phage tail particles could be demonstrated with the “in situ lysis technique” at 60 min following induction. Occasionally, phage heads were demonstrable, but in no instance were complete phage particles discernible. Dividing cells of the bA+ strain of S. marcescens maintained their rod-form following induction with MC until intracellular phage tail bacteriocin particles were seen. However, at 120 min after induction, the swollen, sphaeroplast-like cells lysed, an event that could be correlated with fine structural alterations of the cell wall.
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  • 88
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    Archives of microbiology 126 (1980), S. 277-283 
    ISSN: 1432-072X
    Keywords: Bacteriophage ; Myxococcus ; λ ; Superooiled DNA ; Cross-linking ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract DNA was partially released from the heads of myxococcus phages and also coliphage λ and examined by electron microscopy by a modification of the Kleinschmidt technique, in which water was used as hypophase. DNA emerged from the heads in patterns suggestive of newly relaxed supercoils. The unreleased DNA appeared to occupy discrete regions in the head. Some closed circles were released from λ heads. When NaCl solution was used as hypophase, the DNA was observed either released from the tail or from the head, in the latter case, supercoiled regions were observed. When NH4OAc solution was used as hypophase, tightly wound structures were released from λ heads; these fields also contained supercoiled circles. The presence of constrained supercoiled domains in newly released phage DNA was confirmed by observing the effects of ethidium bromide on its conformation. Treatment of phage with nitrogen mustard, a bifunctional alkylating agent, preserved supercoiled domains, even when the phage were lysed over water as hypophase. Further experiments suggested that phage inactivation by nitrogen mustard is largely due to restraint of the supercoiled, native, tertiary structure and that DNA-protein cross-linking may be involved in this reaction. The implications of these findings for the conformation of phage DNA in vivo are discussed and a new model for the winding of DNA in phage heads is proposed.
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  • 89
    ISSN: 1432-072X
    Keywords: Allomyces ; Zoospores ; Cell wall ; Chitin ; Gamma particle ; Encystment ; Electron microscopy ; Calcofluor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Structural changes during cell wall formation by populations of semisynchronously germinating zoospores were studied in the water mold Allomyces macrogynus. Fluorescence microscopy using Calcofluor white ST (which binds to β-1,4-linked glycans) demonstrated that Calcofluor-specific material was deposited around most cells between 2–10 min after the induction of encystment (beginning when a wall-less zoospore retracts its flagellum and rounds up). During the first 15 min of encystment there was a progressive increase in fluorescence intensity. Ultrastructural analysis of encysting cells showed that within 2–10 min after the induction of encystment small vesicles 35–70 nm diameter were present near the spore surface, and some were in the process of fusing with the plasma membrane. The fusion of vesicles with the zoospore membrane was concomitant with the appearance of electron-opaque fibrillar material outside the plasma membrane. Vesicles similar to those near the spore surface were found within the gamma (γ) particles of encysting cells. These particles had a crystalline inclusion within the electron-opaque matrix. During the period of initial cyst cell wall formation numerous vesicles appeared to arise at the crystal-matrix interface. Approximately 15–20 min was required for the cell wall to be formed. We suggest that the initial response of the zoospore to induction of encystment is the formation of a cell wall mediated by the fusion of cytoplasmic vesicles with the plasma membrane.
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  • 90
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    Archives of microbiology 129 (1981), S. 129-134 
    ISSN: 1432-072X
    Keywords: Cockroach ; Hindgut ; Distribution ; Microbial morphotypes ; Transmission ; Electron microscopy ; Statistical analysis ; Eublaberus posticus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The relative numbers of fourteen microbial morphotypes in transmission electron micrographs of the hindgut of a cockroach, Eublaberus posticus, were counted and their distribution was analyzed statistically. The microbiota of three wall-associated regions (the anterior paunch, the posterior paunch, and the black band region) was clearly different from that of the gut lumen. The three wall fractions were also significantly different from each other. Only one of the fourteen types, prosthecate bacteria, appeared to be distributed randomly in the four fractions. The five main wall-associated morphotypes individually constituted up to 41% of the microbes in some micrographs. They included one type with the characteristic morphology of Methanospirillum. Six morphotypes rarely made up over 2% of the population, but were consistently present. The numbers of the remaining three morphotypes were quite variable between micrographs and between individual insects, but when present often made up 5–10% of the population.
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  • 91
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    Archives of microbiology 133 (1982), S. 97-99 
    ISSN: 1432-072X
    Keywords: Cyanobacteria ; Thylakoid centers ; Photosynthetic membranes/thylakoids ; Membranes ; Membrane biogenesis ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An ultrastructural study of four cyanobacteria (Anabaena cylindrica, Dermocarpa violaceae, Gleocapsa alpicola, Pleurocapsa minor) indicates the presence of previously undescribed thylakoid centers from which photosynthetic membranes (thylakoids) radiate. These peripherally located thylakoid centers are cylinders 30 nm wide by 320 nm long, consisting of globular subunits oriented in nonparallel stacked arrays. Thylakoids are attached to the outer surface of the cylinder along its longitudinal axis. Thylakoid centers appear to be functionally significant due to their structure, location and thylakoid association.
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  • 92
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    Archives of microbiology 130 (1981), S. 339-343 
    ISSN: 1432-072X
    Keywords: Leptospira ; Halophilic ; Electron microscopy ; X-ray analysis ; Inclusions ; Cytoplasmic tubules
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The morphology of cells of strain Muggia, a slightly halophilic leptospire, was examined by the negative staining technique. The ultrastructure of the cells was rather similar to that of cells of Leptonema illini, i. e. the cells possessed cytoplasmic tubules. The basal complex of their flagella, however, was similar to the corresponding part of flagella on Gramnegative bacteria. The interior of the cells was densely packed with inclusions, except for the two outermost wavelengths at each end where these inclusions were absent. X-ray microanalysis showed that the inclusions contained sodium and chlorine as their main constituents. The inclusions disappeared upon storage of the cultures at room temperature.
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  • 93
    ISSN: 1432-072X
    Keywords: Ribulose bisphosphate carboxylase ; Quaternary structure ; Molecular weight ; Electron microscopy ; Cyanobacteria ; Synechococcus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Ribulose bisphosphate (RuP2) carboxylase from the marme cyanobacterium, Synechococcus sp., comprised both large (57,000 dalton) and small (12,000 dalton) subunits. The undissociated, purified enzyme was considerably smaller than the spinach enzyme when compared by pore-gradient electrophoresis, gel filtration and density-gradient centrifugation. This suggested that the cyanobacterial enzyme might have a hexameric (L6S6) subunit structure, unlike the enzymes from spinach and many other organisms which are octamers (L8S8). However, the molecular weight of the Synechococcus enzyme was measured by equilibrium sedimentation and found to be 530,000, which is within the range observed for L8S8-type enzymes. Furthermore, electron microscopic studies of negatively stained preparations of both the native enzyme, and a preparation depleted of 87% of its small subunits by repeated mild-acid precipitation, revealed four-fold symmetry characteristic of an octameric, cubical structure. Synechococcus RuP2 carboxylase therefore must be an L8S8 octamer and its anomalous pore-penetration behaviour may be due to an asymmetric shape. Some support for the latter possibility was provided by electron miscoscopic observations of two different types of images which may be different views of the molecule in two planes.
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  • 94
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    Archives of microbiology 141 (1985), S. 85-90 
    ISSN: 1432-072X
    Keywords: C. sporosphaeroides ; Citrate lyase ; Regulation ; Purification ; Properties ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cells of Clostridium sporosphaeroides which were grown on citrate contained citrate lyase and citrate lyase acetylating enzyme, but no detectable citrate synthase and citrate lyase deacetylase activities. Citrate lyase from C. sporosphaeroides was purified to homogeneity as judged by polyacrylamide gel electrophoresis and high performance liquid chromatography. In contrast to the enzyme from Clostridium sphenoides, the addition of l-glutamate was not necessary for activity and stabilization of the enzyme. The purified enzyme had a specific activity of 34 U/mg protein and was comparable to other citrate lyases with respect to its molecular weight and subunit composition. Electron microscopic investigations showed that similar to the lyase from C. sphenoides and in contrast to all other citrate lyases examined so far, the majority of the enzyme molecules was present in “star” form.
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  • 95
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    Archives of microbiology 142 (1985), S. 259-261 
    ISSN: 1432-072X
    Keywords: Methanogenic bacteria ; Plasmid isolation ; Alkaline lysis ; CsCl gradient ; Restriction endonuclease mapping ; Electron microscopy ; DNA homology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Of 21 recently isolated strains of methanococci, one was found to harbor a small, cryptic, low copy number plasmid. Reproducible recovery was achieved by alkaline lysis of cells pretreated with proteinase K in an osmotically stabilizing buffer. The plasmid was found to contain a singleAval site. No homology was detected between the plasmid and DNA from any of the other new strains or from five known species of methanococci.
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  • 96
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    Archives of microbiology 143 (1986), S. 400-402 
    ISSN: 1432-072X
    Keywords: E. coli relA +/relA ; Starvation survival ; Guanosine tetraphosphate ; Electron microscopy ; Glycogen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Amino acid-starved cells of Escherichia coli relA +, which contain a large number of glycogen particles, are able to survive in phosphate buffer for a longer time period than their relaxed counterparts. With regard to NH 4 + starvation differences in the survival of both strains were not found. NH 4 + starved cells of E. coli relA are able to synthesize glycogen but amino acid-starved cells of the relA strain are not. We suggest that the synthesis of glycogen triggered by guanosine tetraphosphate during amino acid starvation is responsible for the prolonged viability of the E. coli relA + strain.
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  • 97
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    Archives of microbiology 106 (1975), S. 195-200 
    ISSN: 1432-072X
    Keywords: Trichophyton terrestre ; Trichophyton rubrum ; Hyphal fusions ; Origin of intra-hyphal hyphae ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cell observation chamber was designed to perform continuous photomicroscopic observations of hyphal anastomosis and the origin of intra-hyphal hyphae in Trichophyton terrestre and T. rubrum. These data were correlated with ultrastructural features of intra-hyphal hyphae. Hyphal fusions occurred commonly in either species of Trichophyton when incubated alone. In T. terrestre, empty hyphal segments adjoined by live units were invaded at the septa from both directions by new hyphal ingrowth. Continuous observations revealed that the intra-hyphal hyphae subsequently anastomosed via a lateral fusion peg. Similar intra-hyphal hyphae were shown in T. rubrum. Electron microscopic studies revealed ascomycetous septa in both conventional hyphae and intra-hyphal hyphae. For the latter, the cytoplasm and wall of the inner hypha were bounded by cytoplasmic organelles and another cell wall of the outer hypha.
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  • 98
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    Archives of microbiology 163 (1995), S. 300-306 
    ISSN: 1432-072X
    Keywords: Key wordsNitrosomonas europea ; Hydroxylamine ; oxidoreductase (HAO) ; Electron microscopy ; Electron ; spectroscopic imaging ; Quaternary structure
    Source: Springer Online Journal Archives 1860-2000
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    Notes: Abstract The hydroxylamine oxidoreductase from Nitrosomonas europaea was prepared to apparent electrophoretic homogeneity. Electron microscopy of negatively stained preparations of the sample revealed an overall diameter of about 8.8 nm of the enzyme particle. The native structure was determined as a tetrahedron-like assembly of identical subunits exhibiting four protein masses.
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  • 99
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    Archives of microbiology 146 (1986), S. 267-274 
    ISSN: 1432-072X
    Keywords: Photosynthesis ; Membrane structure ; Electron microscopy ; Ectothiorhodospira ; Serial thin sectioning ; Three dimensional reconstruction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The three dimensional organization of the complete photosynthetic apparatus of the extremely halophilic, bacteriochlorophyll b containing Ectothiorhodospira halochloris has been elaborated by several techniques of electron microscopy. Essentially all thylakoidal sacs are disc shaped and connected to the cytoplasmic membrane by small membraneous “bridges”. In sum, the lumina of all thylakoids (intrathylakoidal space) form one common periplasmic space. Thin sections confirm a paracrystalline arrangement of the photosynthetic complexes in situ. The ontogenic development of the photosynthetic apparatus is discussed based on a structural model derived from serial thin sections.
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  • 100
    ISSN: 1432-072X
    Keywords: Thiobacillus ferrooxidans ; Sulfur production ; Sulfur oxidation ; Inhibitors ; Uncouplers ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The intermediary production of elemental sulfur during the microbial oxidation of reduced sulfur compounds has frequently been reported. Thiobacillus ferrooxidans, an acidophilic chemolithoautotroph, was found to produce an insoluble sulfur compound, primarily elemental sulfur, during the oxidation of thiosulfate, trithionate, tetrathionate and sulfide. This was confirmed by light and electron microscopy. Sulfur was produced from sulfide by an oxidative step, while the production from tetrathionate was initiated by a hydrolytic step, probably followed by a series of chemical reactions. The oxidation of intermediary sulfur was severely inhibited by sulfhydryl-binding reagents such as N-ethylmaleimide, by the addition of uncouplers or after freezing and thawing of the cells, which probably damaged the cell membrane. The mechanisms behind these inhibitions have not yet been clarified. Finally, it was observed that elemental sulfur oxidation by whole cells depended on the medium composition. The absence of sulfate or selenate reduced the sulfur oxidation rate.
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