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  • Bone  (222)
  • Autoradiography  (122)
  • Springer  (342)
  • MDPI Publishing
  • 1975-1979  (124)
  • 1970-1974  (218)
  • 1940-1944
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Calcified tissue international 27 (1979), S. 177-185 
    ISSN: 1432-0827
    Keywords: Calcification ; Bone ; Glycoprotein ; Golgi ; Osteoblasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The elaboration of bone matrix glycoprotein by osteoblasts of alveolar bone was investigated by radioautography after the intravenous injection of3H-fucose into young rats. At selected times after injection, animals were sacrificed by intracardiac perfusion and demineralized specimens were prepared for light and electron microscope radioautography. At 5 and 10 min after injection, when the blood fucose level was high, silver grains were restricted to the spheroidal and cylindrical saccules of the Golgi apparatus. At 20 min membrane-limited secretory granules were also labeled. By 35 min, the blood fucose level had dropped and silver grains were detected over the apical cortical cytoplasm, in association with secretory granules located therein. Some grains were present over osteoblast processes and the adjacent prebone matrix. By 4 h most of the silver grains had left the cell. At that time they were observed over prebone, adjacent to osteoblast processes, as well as over the prebone-bone junction where a distinct band of label was noted. In demineralized preparations an electron-dense granular material was present at the prebone-bone junction in association with collagen fibrils. These findings provide evidence that osteoblasts in alveolar bone synthesize fucose-containing glycoprotein and indicate that the addition of3H-fucose occurs in the Golgi apparatus. The glycoprotein passes to the apical cortical cytoplasm by way of membrane-limited secretory granules, is exteriorized, and accumulates at the site where prebone transforms into bone (the prebone-bone junction). Since this is also the site of the calcification front, the deposition of labeled glycoprotein may be related to the deposition of bone mineral.
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  • 2
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    Calcified tissue international 27 (1979), S. 95-99 
    ISSN: 1432-0827
    Keywords: Bone ; Fluoride ; Analysis ; Food ; Age
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Recently published bone fluoride values from Iowa are very high compared to earlier reports, suggesting an increase in fluoride intake. Reanalysis of the Iowa specimens shows levels one-fourth those reported by the Iowa laboratory indicating an error in the original report. Seventeen bone specimens, collected from long-term residents of Rochester, New York, drinking 1 ppm F− water, had a mean value of 2085±270 ppm F− on an ashed-weight basis. This value is not significantly different from that predicted by the data of Zipkin et al. in 1958. These data, therefore, do not support the contention that there has been an increase in fluoride intake.
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  • 3
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    Calcified tissue international 28 (1979), S. 103-105 
    ISSN: 1432-0827
    Keywords: Bone ; Photodensitometry, X-ray ; Radiography ; Spectrophotometry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary A conventional spectrophotometer was used to do radiographic photodensitometry of bone mass. An experiment to produce osteoporosis in guinea pigs required quantification of changes in bone mass. Lack of a densitometer necessitated finding another method for quantification. The method developed involved using industrial film in a closed X-ray system. Optical density of X-rays was then read in a spectrophotometer at nine femur locations. Highly significant correlations between optical density and femur calcium content and femur dry weight per millimeter were obtained. Histological sections from an animal with low optical density measurements confirmed bone loss by showing decreased cortical width and loss of trabecular bone. In addition to using readily available equipment, this method offers the advantage of being able to detect bone loss in specific areas of the bones. In addition, bones are left intact and can be used for other purposes.
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  • 4
    ISSN: 1432-0827
    Keywords: ESR ; Bone ; Enamel ; CO3-Apatite ; Paramagnetism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Carbonate-containing hydroxyapatite, enamel, and bone were irradiated by an X-ray and investigated between 77° and 350°K by means of electron spin resonance (ESR) spectroscopy. The ESR spectrum of enamel irradiated at 77°K in vacuum and observed at the same temperature was almost the same as that of the carbonate-containing hydroxyapatite. The temperature dependence of signal intensities confirms a spin-energy exchange between the mineral and organic constituents in bone, but in enamel no or very little spinenergy exchange between the mineral and organic constituents. Considerable similarity among the ESR spectra of enamel, bone, and carbonate-containing apatite was obtained after X-ray irradiation in air at 300°K with both an X-band and a Q-band ESR spectrometer. The Q-band spectrum can be interpreted in terms of two paramagnetic species. One is identified as a CO 3 3− anion radical which has an axial symmetry withg factors of 2.0029 and 1.9972. The other species is found to be centered atg=2.0019.
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  • 5
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    Calcified tissue international 28 (1979), S. 33-36 
    ISSN: 1432-0827
    Keywords: Infrared, visible light ; Bone ; Collagen ; Apatite
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Photovoltaic effect (infrared and visible light) is observed in bone and its two major components, collagen and apatite, at room temperature. A dimunition in the magnitude of photovoltage is observed after exposure to ultraviolet light in all the cases. The drift mobility of the charge carriers is obtained by measuring I versus V relationships in sandwich samples and relating them to the permitivity of the medium. Lifetime of the injected carriers is measured in the usual way. The data are consistent with the hypothesis that the effects are due to protonic conduction phenomena.
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  • 6
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    Development genes and evolution 185 (1979), S. 295-303 
    ISSN: 1432-041X
    Keywords: Myogenesis ; Cell cycle ; Autoradiography ; Cytophotometry ; Bombina
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary InBombina variegata, striated myofibrils first appear in G2 uninucleated primary myoblasts. Multinucleated muscle fibres form later as a result of the fusion of primary myobasts with secondary myoblasts of mesenchymal origin. The nuclei of the polykaryocytes vary in size and DNA content (nuclear dimorphism). The larger nuclei of the primary myoblasts retain tetraploid quantities of DNA, whereas the smaller nuclei of the secondary myoblasts are diploid. From this we conclude that fusion can take place between cells that are in different phases of the cell cycle (G1–G2). Our findings are compared with those on myogenesis in other chordate species and are confronted with the current commonly accepted model of vertebrate muscle differentiation.
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  • 7
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    Calcified tissue international 27 (1979), S. 33-40 
    ISSN: 1432-0827
    Keywords: Chick embryo ; Bone ; Organ culture ; Scanning electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The study describes the ultrastructure of the mineralized portion of chick tibiae from 10 days in ovo to 2 days post-hatch. At 10 days a single mineralized cylinder surrounds the diaphysis. On its outer surface columnar trabeculae join to form ridges parallel to the long axis of the bone. These ridges are covered by another cylinder and form the haversian canals. At 11 days vascular invasion of the marrow cavity occurs and resorption of the endosteal surface begins. This type of periosteal deposition and endosteal resorption is repeated during and subsequent to embryonic development. The mineralized portion of 10-day chick tibiae cultured for 2 days in modified BGJ medium was compared with 10-, 11-, and 12-day tibiae in ovo. Cultured tibiae were similar in length and calcium content to 11-day tibiae in ovo. The form of mineral deposited in ovo and in culture was the same, namely, aggregates of spherical mineral clusters. Differences in culture included the following: (a) few concentric cylinders were deposited as compared with tibiae in ovo; (b) trabeculae were not arranged in rows and ridges in culture; (c) osteocytic lacunae were restricted to bases of trabeculae rather than uniformly distributed as in ovo; and (d) the endosteal surface of tibiae in culture appeared etched.
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  • 8
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    Calcified tissue international 27 (1979), S. 57-64 
    ISSN: 1432-0827
    Keywords: Bone ; Bone formation ; Calcification ; Calcification nodule ; EM
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Osteolathyrism has been used as an experimental model for the study of calcification nodules during the mineralization process. Periosteal exostoses developing in osteolathyrism characteristically have spherical basophilic structures (calcification nodules) in the vicinity of developing bone spicules. In thin sections, the nodules were seen scattered between collagen fibers in the intercellular matrix. Collagen fibers did not appear to be present within the nodules but sometimes were packed just outside them. Matrix vesicles were also present in areas of early mineralization. After EDTA decalcification, the majority of the nodules consisted of a fine granular material surrounded by an electron-dense peripheral zone. The peripheral dense zone was occasionally incomplete in small nodules in areas of early mineralization. An electron-dense central area could be observed in the center of the nodules. Evidence has been presented indicating that the calcification nodules arise from smaller mineralization foci, presumably matrix vesicles. The calcification nodules enlarge to approximately 1.0 µm in size, at which point development is slowed or halted allowing the formation of the peripheral dense zone. Although coalescence of nodules was observed, this was more a random event. The further mineralization of the trabeculae was achieved by the calcification of the collagen fibers. The mineralized trabeculae reflected this pattern of nodular and collagenous calcification. It is suggested that this pattern of calcification is characteristic of rapidly developing woven bone.
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  • 9
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    Calcified tissue international 27 (1979), S. 105-108 
    ISSN: 1432-0827
    Keywords: Bone ; Femur ; Strength ; Density ; Age
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary In order to ascertain whether the intrinsic strength of human bone changes with age or not, we have determined the ultimate tensile strength and density of strips of femoral cortical bone. These femora were collected from cadavers varying in age from 13 to 97 years. The results show that both density and intrinsic strength of bone increase up to about the fourth decade of life and then decrease with age. However, the rate of decrease of strength is greater than that of density. This indicates that the density of bone is not the sole determining factor of its strength, and that some other factors play an important part.
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  • 10
    ISSN: 1432-0827
    Keywords: Bone ; Rickets ; Hypophosphatemia ; Microradiography ; D-resistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The perilacunar areas of low mineral density in microradiographs from cortical bone of patients with hypophosphatemic (vitamin D-resistant) rickets are not evenly distributed throughout the bone tissue. Their frequency and distribution were determined in bone from 9 patients with this disease. It was found that the lesion was more frequent in haversian bone than in interstitial bone, and along the inner circumference of growing haversian systems as compared with the outer circumference. These findings indicate that the lesion is the result of retarded mineralization, and that mineralization slowly proceeds in these areas as the bone becomes older. A relatively high frequency of the lesion was also found in osteons with an elliptical cross section along the long axis of the ellipse. The cause of the abundance of the lesion at these sites is not clear, but it is possible to explain the uneven distribution in elliptical osteons by assuming an unequal rate of bone formation in these structures.
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  • 11
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    Calcified tissue international 27 (1979), S. 165-169 
    ISSN: 1432-0827
    Keywords: Oxygen uptake ; Bone ; Oxygen tension ; Carbon dioxide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The present study was carried out to determine the influence of carbon dioxide and oxygen tension on the respiratory activity of bone cells in mouse calvaria in vitro. Five-day-old mouse calvaria were removed aseptically and incubated individually for 1 h at 37° C in a closed reaction chamber containing 1.5 ml of tissue culture medium made up of 60% horse serum in Gey's solution containing 100 unit/ml penicillin and 100µg/ml streptomycin. Before the calvaria were added, the medium in the incubation chamber was equilibrated with 10%, 20%, 30%, or 50% oxygen balanced with nitrogen. The effect of CO2 on oxygen utilization by the calvaria was determined by incubating the calvaria in a medium previously equilibrated with either 50% O2 balanced with N2 or 50% O2 and 5% CO2 balanced with N2. At each oxygen tension, the rate of oxygen utilization by the calvaria was measured polarographically by a Clark oxygen electrode. The results showed that the rate of oxygen uptake of bone increased as the oxygen tension increased and carbon dioxide stimulated significantly the rate of oxygen utilization by the bone cells. In view of the previous reports that both carbon dioxide and oxygen tension are implicated in the process of bone resorption, it is suggested that these two factors may affect bone resorption by influencing the oxygen utilization by bone cells and ultimately controlling their energy metabolism.
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  • 12
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    Calcified tissue international 28 (1979), S. 37-42 
    ISSN: 1432-0827
    Keywords: X-ray diffraction ; Hydroxyapatite ; Whitlockite ; Fish (Pisces) ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Pyrolyzed scales, fin spines, and bone from the ray-finned bony fishPolypterus (Actinopterygii) showed two mineral phases on X-ray diffraction: hydroxyapatite (HA), Ca5(PO4)3OH, and whitlockite, Ca3(PO4)2. The ratio of HA/whitlockite varied with the structure (scale, spine, bone) within each individual fish. The relative proportions of HA to whitlockite in pyrolyzed samples reflected the Ca/P ratio of the sample. Whitlockite appears after pyrolysis when the Ca/P is lower than 1.67. Among the five fish investigated, for each structure a general trend was noted. The proportion of HA relative to whitlockite increased with size (age) of the fish. Thus the smallest fish, a juvenile, exhibited a low Ca/P mineral in its calcified tissues, whereas the larger fish had progressively more HA and less whitlockite.
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  • 13
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    Calcified tissue international 28 (1979), S. 17-22 
    ISSN: 1432-0827
    Keywords: Anticonvulsant ; Ketogenic diet ; Calcium ; Vitamin D ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Vitamin D and mineral metabolism status was examined in five children maintained chronically on combined ketogenic diet-anticonvulsant drug therapy (KG), and the results compared to those obtained in 18 patients treated with anticonvulsant drugs alone (AD) and 15 normal controls. KG patients exhibited biochemical findings of vitamin D deficiency osteomalacia: decreased serum 25-hydroxyvitamin D (25OHD) and calcium concentrations, elevated serum alkaline phosphatase and parathyroid hormone concentrations, decreased urinary calcium and increased urinary hydroxyproline excretion, and decreased bone mass. Although the KG and AD groups demonstrated similar reductions in serum 25OHD concentration, the KG patients exhibited a significantly greater reduction in bone mass. In response to vitamin D supplementation (5000 IU/day), mean bone mass in the KG group increased by 8.1±0.9% (P〈0.001) over a 12-month period. These results suggest that ketogenic diet and anticonvulsant drug therapy have additive deleterious effects on bone mass and that these effects are partially reversible by vitamin D treatment.
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  • 14
    ISSN: 1432-0827
    Keywords: Bone ; Bone cells ; Cyclic AMP ; Osteoclast activating factor ; Lymphokine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Osteoclast activating factor (OAF) is a lymphokine which may participate in the pathologic destruction of bone observed in a number of disorders. In the current studies, we investigated the action of OAF on cAMP accumulation by bones and isolated bone cells in culture. OAF was shown to stimulate accumulation of cAMP in mouse cranial bones at doses between 1 and 1000 ng/ml. Stimulation of bone resorption was observed in bones treated with the same doses of OAF. In order to investigate the cell types responsible for cAMP responses to OAF, we isolated bone cells and grew them in monolayer culture. The cells were isolated by a variety of techniques which separate bone cells into two types of parathyroid hormone (PTH)-responsive populations: (a) cells derived from the periosteal regions of the bone, which also respond to calcitonin with increases in cAMP; and (b) cells derived from the matrix, which do not respond to calcitonin. OAF caused elevation of cAMP levels in both the periosteum-derived cells and the matrix-derived cells. The magnitudes and time courses of OAF effects in these populations resembled the effects previously reported for PTH in the same populations. OAF stimulated adenyl cyclase in both types of cell populations, but did not produce significant changes in cAMP phosphodiesterase activity. OAF differed from PTH in that its effects on cAMP accumulation decreased sharply at supramaximal doses in both bone and isolated cells, especially in the matrix-derived populations. Bone resorption did not decrease as markedly as did cAMP accumulation at high doses of OAF, suggesting that cAMP accumulation and resorption could be dissociated under some conditions. These results indicate that OAF has effects on cAMP production in the same cell populations as PTH, and suggest that OAF could modify not only resorption but also formation of bone in vivo. OAF may exert its effects on bone by means of cAMP-dependent mechanisms, but more data will be necessary to establish this unequivocally. The observed differences between OAF and PTH may be of relevance in the mechanism and treatment of pathologic bone destruction in vivo.
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  • 15
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    Calcified tissue international 27 (1979), S. 199-204 
    ISSN: 1432-0827
    Keywords: Bone ; Parathyroid hormone ; Calcitonin ; Osteoporosis ; Paraplegia ; Hydroxyproline
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary In paraplegia, osteoporosis below the neurological lesion occurs early after the spinal cord affection. The serum levels of parathyroid hormone (PTH) and calcitonin (CT), using a radioimmunoassay for the measurement of immunoreactivity, were studied in 12 paraplegic patients for 9 months following onset. Serum Ca and P levels, urinary hydroxyproline excretion, and the kinetic metabolic clearance of45Ca have also been measured. P and immunoreactive (i) CT levels were found the highest at the beginning of the observation and progressively decreased with time. Ca and iPTH serum levels varied inversely with time, the highest level of Ca and the lowest level of iPTH being recorded at the third month following the paraplegia. Mean values of Ca, iPTH, and iCT were in the normal range throughout the study. P levels were increased during the first 3 months. Hydroxyprolinuria was also high and45Ca kinetics showed increased values of Vt, Vo+, and Vu. These parameters indicate a high degree of bone turnover. The results were consistent with the assumption that PTH is not responsible for the increased resorption of bone in paraplegia. Likewise, a deficiency of CT does not seem to be responsible for this bone resorption. These endocrine modifications could be secondary to an increase in the calcium flux from bone to blood and resulting from bone destruction as attested by the increase of urinary calcium and urinary hydroxyproline excretion.
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  • 16
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    Calcified tissue international 27 (1979), S. 233-237 
    ISSN: 1432-0827
    Keywords: Bone ; Vitamin D ; Acidosis ; Phosphate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Vitamin D and phosphate deficiency were produced in rats in order (a) to evaluate the degree of bone mineral and matrix maturation using a bromoform/toluene density gradient technique; and (b) to compare the aforementioned bone maturational changes due to vitamin D and phosphate deprivation to those produced with superimposed severe acidosis. Rats were fed a diet deficient in vitamin D and phosphorus (0.2%) from 3 weeks through 7 weeks of age. To examine the additional contribution of dietary calcium, we gave one-half of the animals either a low (0.06%) or high (1.3%) calcium diet. Following the 4 weeks of vitamin D deficiency, one-half of each group was given 1.8% NH4Cl in the drinking water for 4 succeeding days to induce an acute, severe acidosis. The degree of bone maturation was quantitated via bromoformtoulene density gradient fractionation; total mineral and hydroxyproline (collagen) levels were quantitated as well. The vitamin D-deficient rats deprived of adequate dietary phosphate responded by conserving phosphorus, and as a consequence total bone phosphorus levels were maintained within that level for control rats. This conservation was independent of calcium intake but was extremely sensitive to acute acid loading, where a significant reduction in total bone phosphorus was noted. The bone maturational profile obtained from the vitamin D-phosphate deficient rats, however, revealed a significant accumulation of less mature or dense bone collagen and mineral with a corresponding decrease in the most mature or dense moieties. In contrast to the reduction of the total bone phosphorus content by acute acidosis, the skeletal collagen-mineral maturational profile was not significantly affected by the short-term systemic acidosis. The observed retardations in the bone collagen and mineral maturation of the vitamin D-deficient, phosphate-deprived state provide an additional observation which may well relate to the progressive osteopenia documented in states of chronic, mild acidosis.
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  • 17
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    Calcified tissue international 27 (1979), S. 275-279 
    ISSN: 1432-0827
    Keywords: Proline ; Cartilage ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Proline biosynthetic and degradative enzymes are unevenly distributed in differentiated mammalian tissues. Activities of the synthetic enzymes are relatively high in collagenous tissues, whereas activities of the degradative enzymes are high in noncollagenous tissues. In order to further characterize tissue-specific proline biosynthesis and degradation, we have determined proline enzyme activities during cartilage and bone formation induced by demineralized bone matrix. We can thus follow temporal changes in enzyme activity in a single tissue as different cell types develop. Ornithine aminotransferase and pyrroline-5-carboxylate reductase have peaks of activity which correlate with maximal type II collagen synthesis by chondrocytes. Both enzymes also are active during bone formation. In contrast, proline oxidase and pyrroline-5-carboxylate dehydrogenase are present at low levels and do not change as new cell types appear. Arginase activity peaks during the first 3 days and then rapidly decreases by the time cartilage and bone formation begin. These observations further substantiate the importance of proline biosynthesis in collagenous tissues. The close correlation between ornithine aminotransferase activity and type II collagen synthesis suggests that the pathway from ornithine to proline may be especially important during formation of type II collagen.
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  • 18
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    Calcified tissue international 29 (1979), S. 41-46 
    ISSN: 1432-0827
    Keywords: Albumin ; Dentine ; Enamel ; Autoradiography
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Rabbit serum albumin labeled with125I was injected into rabbits (650–870 g). The incorporation pattern of the labeled albumin into the developing dentine and enamel matrix was studied 1 h, 6 h, and 3 days after the injection using autoradiography. Silver grains were observed over the predentine 1 h after the injection. They were found over the dentine matrix adjacent to the predentine-dentine junction at 6 h. The silver grain band was concentrated over the internal dentine matrix at 3 days. However, no appreciable amount of the silver grains could be seen over the enamel matrix at any time after the injection. It is possible that there might be some physiological barrier between the extravascular fluid and the enamel matrix. The role of the albumin in dentine is discussed.
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  • 19
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    Calcified tissue international 27 (1979), S. 109-115 
    ISSN: 1432-0827
    Keywords: Bone ; Bending rigidity ; Disuse atrophy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The in vivo bending rigidity and bone mineral content of monkey ulnae and tibiae were measured. Bending rigidity in the anteroposterior plane was measured by an impedance probe technique. Forced vibrations of the bones were induced with an electromechanical shaker, and force and velocity at the driving point were determined. The responses over the range of 100–250 Hz were utilized to compute the bending rigidity. Bone mineral content in the cross section was determined by a photon absorption technique. Seventeen male monkeys (Macaca nemestrina) weighing 6–14 kg were evaluated. Repeatability of the rigidity measures was 4%. Bone mineral content was measured with a precision of 3.5%. Bending rigidity was correlated with the mineral content of the cross section,r=0.899. Two monkeys were evaluated during prolonged hypodynamic restraint. Restraint produced regional losses of bone most obviously in the proximal tibia. Local bone mineral content declines 17 to 24% and the average bending rigidity declines 12 to 22%. Changes in bones leading to a reduction in mineral content and stiffness are discussed.
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  • 20
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    Calcified tissue international 27 (1979), S. 13-18 
    ISSN: 1432-0827
    Keywords: Anticonvulsants ; Calcium ; Vitamin D ; Parathyroid hormone ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Parameters of mineral metabolism were examined in 6 patients with moderately severe anticonvulsant drug-induced osteomalacia. Compared to 15 matched controls, the patients exhibited significantly reduced serum calcium, inorganic phosphate, and 25-hydroxyvitamin D concentration, elevated serum alkaline phosphatase and immunoreactive parathyroid hormone (iPTH) concentration, reduced intestinal47Ca absorption, reduced urinary calcium and increased urinary hydroxyproline excretion, and reduced forearm bone mass. Intestinal absorption of vitamin D3 was normal. Following 4 months of treatment with vitamin D3 (4000 units/day), serum 25-OHD concentration was increased to 3 times mean normal values and all parameters except serum iPTH, urinary calcium excretion, and forearm bone mass were returned to levels not significantly different from normal. Serum iPTH concentration was reduced by 39% (P〈0.05); 24-h urinary calcium excretion rose by 98% (P〈0.001), and forearm bone mass increased by 5.6% (P〈0.05). It is concluded that moderate-dose vitamin D3 supplementation is effective in normalizing parameters of mineral metabolism in this disorder, despite evidence of resistance to the biologic effects of vitamin D.
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  • 21
    ISSN: 1432-0827
    Keywords: Calcium ; PTH ; 1,25(OH)2D ; Acid ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary We measured mineral and acid balances, serum iPTH, urinary cAMP/creatinine, and plasma concentrations of 25OHD and 1,25(OH)2D in 7 healthy adults during control conditions and during increased fixed acid production achieved either by the administration of NH4Cl (N=3) or by increased dietary protein intake (N=4). When acid production was increased, the subjects were in positive acid balance and negative Ca balance because of increased urinary Ca excretion. Serum iPTH fell slightly but urinary cAMP and the plasma levels of vitamin D metabolites did not change. We conclude that the accelerated skeletal and urinary losses of Ca that occur when fixed acid production is increased are not contributed to nor compensated for by the parathyroid-vitamin D endocrine systems.
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  • 22
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    Calcified tissue international 29 (1979), S. 169-171 
    ISSN: 1432-0827
    Keywords: Glycosaminoglycans ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Compact bone tissue chondroitin sulphate, previously considered to be pure chondroitin-4-sulphate, was isolated from three adult human femoral diaphyses and digested with chondroitinase ABC. Assaying these digests by means of high performance liquid chromatography we could demonstrate that 12 to 14% of the disaccharide residues were 6-sulphated. The 4/6-ratios were also studied in chondroitin sulphate fractions of different molecular size. Slightly increasing amounts of 6-sulphated disaccharides were found with increasing size of the chondroitin sulphates.
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  • 23
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    Calcified tissue international 29 (1979), S. 7-13 
    ISSN: 1432-0827
    Keywords: Bone ; Culture ; Resorption ; Acidosis ; Alkalosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary We have examined the effects of H+, CO2, and HCO3 − concentrations during metabolic and respiratory acidosis and alkalosis on bone resorption in vitro. Rat fetal bones prelabeled with45CaCl2 were cultured at 2%, 5%, and 10% CO2 for up to 120 h, and the release of45Ca was measured in devitalized bones (non-cell-mediated45Ca release) and in live bones (cell-mediated45Ca release) cultured with or without PTH and 1,25(OH)2D3. Non-cell-mediated mineral loss was linearly related to H+ concentration but not to CO2 or HCO3 − concentration. This effect was observed on both labeled and stable calcium. Over a wide pH range (6.9–7.5) H+, CO2, or HCO3 − concentrations did not influence cell-mediated bone resorption in control or in PTH-and 1,25(OH)2D3-stimulated cultures. However, inhibition of cell-mediated bone resorption was observed at higher or lower pH irrespective of CO2 or HCO3 − concentrations. These observations demonstrate that the bone mineral mobilizing effect of acidosis in vitro is mainly due to the effect of changing H+ concentration on devitalized bone. Effects on cell-mediated bone resorption and hormonal response were observed only at extremes of pH. The effects of H+ were independent of changes in CO2 or HCO3 − concentration and could be responsible for the negative calcium balance and increased urinary loss observed in metabolic acidosis in vivo, but do not explain the reported differences in effects on calcium metabolism between respiratory and metabolic acidosis.
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    Planta 146 (1979), S. 31-39 
    ISSN: 1432-2048
    Keywords: Autoradiography ; Calcium ; Mougeotia ; Phytochrome
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Ca2+ is proposed to function as a messenger in such phytochrome-mediated responses as localized cell growth, intracellular movements, and control of plasma membrane properties. To test this hypothesis, the uptake of Ca2+ in irradiated and non-irradiated regions of individual threads of the green alga Mougeotia was studied with the aid of 45Ca2+ and low temperature autoradiography: 10–20 cells within 40–60 cell-long threads were irradiated for up to 1 min, transferred to darkness for 3 to 10 min, submersed in a radioactive medium for 1 min, washed in an unlabelled medium for 30 min, and then autoradiographed at-80° C for several days. The autoradiographs show that those cells which had been pre-irradiated with red light did take up 2–10 times more Ca2+ than the adjacent non-irradiated cells of the same thread. Cells pre-irradiated with farred light or red light followed by far-red light showed no enhanced uptake of Ca2+. These results might be interpreted to indicate, firstly, that phytochrome-Pfr is involved in the enhanced uptake of Ca2+ and secondly, that the accumulation of radioactive Ca2+ in red light irradiated cells is an expression of an increased intracellular concentration of Ca2+. This interpretation is based on the data that (i) the dark interval between irradiation and labelling precluded the involvement of photosynthesis, (ii) the effect of red light was reversible with far-red light, and (iii) the accumulation of Ca2+ persisted during the long wash-out period. We speculate, that the red light-enhanced accumulation of Ca2+ in Mougeotia cells is caused by a Pfr-mediated increase of the Ca-permeability of the plasma membrane, and perhaps by a Pfr-impeding of an active Ca2+-extrusion.
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  • 25
    ISSN: 1432-2048
    Keywords: Autoradiography ; Chloramphenicol ; Phaseolus ; Plastids ; Protein synthesis ; Transport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Leaf discs from etiolated bean plants were found to incorporate [3H]lysine into 80 S ribosomesynthesized proteins in the presence of chloramphenicol (100 mg l−1) when exposed to light. After a 7 min pulse of [3H]lysine, the discs were transferred to the same medium but with nonradioactive lysine, and postincubation was carried out for 24 h. The number of silver grains over the plastids, after the first period of a lag phase, indicates a large increase between 12 and 24 h of postincubation. Simultaneously, the labeling of the cytoplasm becomes reduced during that period. The results show that during inhibition of the protein formation within plastids, the synthesis of plastid-destined proteins in cytoplasm, as well as their transport into plastids, can still proceed.
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    Archives of microbiology 123 (1979), S. 129-135 
    ISSN: 1432-072X
    Keywords: Bean rust ; Uromyces phaseoli ; Phaseolus vulgaris ; Autoradiography ; Electronmicroscopy ; Haustorium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To study the function of bean rust haustoria either the host, Phaseolus vulgaris, or the parasite, Uromyces phaseoli, were labeled with 3H-lysine. 24 h and 48 h after inoculation samples of the infected leaves were taken and processed for electronmicroscopic autoradiography. When the host was labeled, silver grain densities over the structures of host and parasite were compared. Very low grain densities were found over developing haustoria of the bean rust and higher grain densities over mature and old haustoria. Compared with mature haustoria, reduced grain densities were observed over intercellular hyphae. It is assumed that a flow of labeled metabolites from the host cell to the haustorium and then to the intercellular hypha took place. When sporelings originating from labeled uredospores infected the leaf, no detectable amount of 3H-lysine or its metabolites were found to be transferred from the intercellular hyphae or haustoria to cells of the leaf parenchyma.
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    Theoretical and applied genetics 54 (1979), S. 277-282 
    ISSN: 1432-2242
    Keywords: Nuclear cycle ; S period ; Zea mays ; Autoradiography
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The nuclear cycle among several diverse genetic stocks of Zea mays root meristem cells was compared and it was found that there were no significant differences among the nuclear cycle durations and its component phases. The durations of various periods of their mitotic cycles were studied by autoradiography of cells pulse-labelled with tritiated thymidine (3H-TdR). The total nuclear cycle was 10 to 11.5 hours and mitosis was 0.81 to 1.34 hours at 25°C. The S period is the longest interval (50% of the total time) of the nuclear cycle; of the rest of the cycle, G2 is longer than G1 or mitosis among all stocks. The constancy of the nuclear cycle among several stocks was adduced as evidence for strict genetic control of the cycle. Furthermore, it is demonstrated the DNA synthesis period is not dependent upon the amount of DNA present.
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    Cell & tissue research 200 (1979), S. 311-322 
    ISSN: 1432-0878
    Keywords: Mouse pituitary ; Autoradiography ; Glucocorticoid ; Antiglucocorticoid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Résumé La localisation au niveau de l'hypophyse de souris de la 3H-dexaméthasone (3HDM), de la 3H-corticostérone (3HB) et de la 3H-cortexolone (3HS) est étudiée par autoradiographie 30 min après l'injection des composés tritiés seuls ou en présence de stéroides compétiteurs non radioactifs. La 3H-dexaméthasone s'accumule dans des noyaux cellulaires de la pars distalis et de la pars nervosa mais pas dans des noyaux de la pars intermédia. Cette accumulation préférentielle (densité des grains d'argent nucléaire/cytoplasmique: 4/1) est abolie par l'injection de 3H-dexaméthasone en présence de dexaméthasone non radioactive. La 3H-corticostérone se concentre avec une intensité beaucoup plus faible que la 3H-dexaméthasone uniquement dans certains noyaux de la pars distalis. Un excès de corticostérone non radioactive diminue la densité nucléaire des grains d'argent des cellules de la pars distalis. Après injection de 3H-cortexolone, aucune accumulation préférentielle de grains d'argent n'est observée dans les noyaux cellulaires. Dans une seconde série d'expériences, la 3H-dexaméthasone est injectée soit en présence d'excès de dexaméthasone (10x, 100x) ou de corticostérone (100x, 300x) ou de cortexolone (100 x, 300 x). Dans ces conditions, la densité cytoplasmique des grains d'argent n'est pas différente de celle observée après injection de 3H-dexaméthasone seule mais l'accumulation préférentielle de la radioactivité dans les noyaux cellulaires est abolie. Des études biochimiques parallèles effectuées sur des préparations cytoplasmiques de rein des mêmes animaux montrent que la radioactivité cytoplasmique totale ne varie pas tandis que la liaison cytoplasmique de la 3H-dexaméthasone diffère suivant les traitements. Ces résultats montrent i) que la dexaméthasone se fixe spécifiquement dans des noyaux cellulaires de la pars distalis et de la pars nervosa et que cette fixation nucléaire est abolie aussi bien par des excés de corticostérone ou de cortexolone que par des excès de dexaméthasone, ii) que la corticostérone se localise dans des noyaux cellulaires de la pars distalis mais beaucoup moins intensément que la dexaméthasone, iii) que la cortexolone se comporte comme un antagoniste des glucocorticoides au niveau de la cellule hypophysaire.
    Notes: Summary Autoradiograms of mouse pituitaries were prepared 30 min after injection of 3H-dexamethasone (3HDM), 3H-corticosterone (3HB) and 3H cortexolone (3HS) either alone or in the presence of competing unlabelled steroids. 3H-dexamethasone accumulated in cell nuclei of both the pars distalis and the pars nervosa but not in those of the pars intermedia. This preferential accumulation (nuclear/cytoplasmic grain density, 4∶1) was abolished by the concurrent administration of excess dexamethasone. 3H-corticosterone, to a much less marked extent than 3H-dexamethasone, accumulated in cell nuclei of the pars distalis but not in those of the pars intermedia and the pars nervosa. Excess unlabelled corticosterone diminished nuclear grain density in the pars distalis. After 3H-cortexolone injection, preferential nuclear uptake was not observed. In a second series of experiments, excess dexamethasone (10 x, 100 x), corticosterone (100 x, 300 x) and cortexolone (100 x, 300 x) administered with 3H-dexamethasone were without effect on cytoplasmic grain density but totally abolished preferential nuclear accumulation. Parallel biochemical studies on kidney cytoplasmic preparations from the same animals showed no differences in total cytoplasmic radioactivity between treatments but marked differences in cytoplasmic bound 3H-dexamethasone. The results demonstrate: i) that dexamethasone binds specifically to cell nuclei of the pars distalis and the pars nervosa and that this nuclear concentration is abolished by competing corticosterone and cortexolone as well as dexamethasone; ii) that corticosterone localizes in cell nuclei of the pars distalis but much less markedly than dexamethasone; iii) that cortexolone fullfils the criteria of a glucocorticoid antagonist at the pituitary cell level.
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    Cell & tissue research 198 (1979), S. 435-440 
    ISSN: 1432-0878
    Keywords: Teleost ; Pituitary gland ; Estrogen ; Gonadotropes ; Autoradiography
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of estradiol-concentrating cells in the pituitary of the platyfish, Xiphophorus maculatus, is studied after the injection of 3H estradiol-17β by thaw-mount autoradiography. Autoradiograms prepared 2–8 h after the injection show nuclear concentration of radioactivity in certain cells of the proximal pars distalis, while no nuclear labeling is found in cells of the rostral pars distalis, pars intermedia and pars nervosa. Radioactively labeled cells are identified as gonadotropes by a combined technique of thaw-mount autoradiography and immunoperoxidase staining with antiserum to ovine LH. Approximately 80% of the immunoreactive LH cells show a concentration of radioactivity in their nuclei. These observations indicate that in teleosts, as in mammals, estradiol has a direct effect on pituitary gonadotropes.
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    Cell & tissue research 201 (1979), S. 249-262 
    ISSN: 1432-0878
    Keywords: Kidney ; Androgens ; 3H-Thymidine ; Autoradiography ; Fish
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Résumé L'incorporation de thymidine tritiée dans les tubules proximaux du rein est étudiée par autoradiographie rapide chez des Epinoches femelles préalablement traitées par la méthyltestostérone. Les résultats sont comparés avec ceux obtenus chez des mâles normaux ayant ou non développé un rein muqueux. Des cellules marquées sont présentes à tous les niveaux des tubules proximaux, mais leur nombre varie considérablement d'un segment à l'autre. Elles sont les plus nombreuses dans la seconde partie du tubule proximal, particulièrement dans sa région distale. Le taux de marquage est modifié dans toutes les régions du néphron, mais les variations sont les plus intenses dans la région distale de la seconde partie du tubule proximal, où l'on observe parfois la formation de nouveaux tubules par bourgeonnement. La comparaison des résultats obtenus dans les deux sexes fait apparaître une différence de réactivité au niveau de la zone proximale de la seconde partie du tubule proximal, où la méthyltestostérone agit fortement chez les femelles, alors qu'elle n'exerce pas d'effet notable chez les mâles. L'augmentation de volume du rein chez le mâle lors de la période de reproduction ne résulte donc pas uniquement d'un gonflement des cellules de la seconde partie du tubule proximal, comme on le pensait généralement. Des phénomènes mitotiques interviennent également dans ce processus, sous l'influence des hormones sexuelles mâles, qui conduisent à un allongement, voire à une multiplication des tubules proximaux.
    Notes: Summary The rate of 3H-thymidine incorporation into different parts of the renal proximal tubule of female sticklebacks treated with methyltestosterone was investigated using high-speed scintillation autoradiography. The results are compared with those from normal males before or after mucous transformation of the kidney. Labelled cells are observed in all parts of the proximal tubule, with marked variations from one segment to another. They are numerous in part 2 of the proximal tubule, particularly in the distal region. Male sex hormones affect the labelling rate in all parts of the nephron, especially in the distal region of part 2 of the proximal tubule. In that particular area, new tubule formation by budding is observed in some individuals, but this process does not appear to be a general one. Correlation between the frequency of these figures and the time of treatment could not be established. Comparing the action of sex hormones in females with that in males reveals a difference in reactivity in the proximal zone of part 2 of the proximal tubule, where methyltestosterone has a strong action in females; in contrast, in “mature” and “immature” males, only a few labelled cells are present in this region. It is concluded that kidney enlargement during the breeding season does not result only from a swelling of cells belonging to part 2 of the proximal tubule, as was generally believed, but also from a lengthening or even a proliferation of the proximal tubules, induced by an increase in mitotic activity controlled by male sex hormones.
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    Cell & tissue research 201 (1979), S. 467-477 
    ISSN: 1432-0878
    Keywords: Anterior pituitary ; Hormone synthesis ; Morphometry ; Autoradiography ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In order to compare the functional state of the anterior pituitary of spontaneously hypertensive rats (SHR) with that of normotensive Wistar Kyoto rats (WKR), the anterior pituitary was examined by morphometry and autoradiography at the level of electron microscopy. The relative number and the relative volume of thyrotrophs in the anterior pituitary were significantly greater in SHR compared with age-matched WKR at 0, 7, 30–33 days, and 10 months of age, while the relative number of somatotrophs in SHR was significantly smaller at 1 and 10 months of age. Electron microscope autoradiographic analysis of uptake of 3H-lysine by thyrotrophs of both strains at the age of approximately one month showed that 3H-lysine was incorporated into protein and transported finally to secretory granules which migrated to near the cell membrane to be discharged. Silver grains were significantly more numerous over the thyrotrophs of SHR than over those of WKR at 30 min, 1 h, and 4h after the injection of 3H-lysine. The present study has ascertained morphologically that a congenital hypersynthesis of TSH by the anterior pituitary occurs in SHR.
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    Plant and soil 51 (1979), S. 551-561 
    ISSN: 1573-5036
    Keywords: Autoradiography ; Culture solution ; Fe deficiency ; Millet ; P/Fe interaction ; Phosphorus ; Sorghum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Seedlings of millet (Pennisetum americanum (L) K. Schum var. Tift 23 DA × Tift 18 DB) and guineacorn (Sorghum bicolor (L) Moench var. RS' 702) were employed in growth chamber studies to evaluate the effects of various phosphorus (P) levels on their growth and nutrient composition. The growth medium was Hoagland solution containing 0.0, 0.1, 1.0 and 2.5 mM P and 2.2 mM ferrous sulphate-tartate solution. At the end of twenty-four days, two plants from each treatment were supplied with 0.31 μci/ml of radioactive iron (55Fe) (with the same P levels used to grow them) and then allowed to absorb the55Fe for 3 hours after which they were harvested, separated into roots and shoots. The two components were wet-ashed separately, and assayed for55Fe. The rest of the plants in Hoagland solution were also harvested, separately into roots and shoots. The components parts were also wet-ashed separately and assayed for iron (Fe), manganese (Mn), zinc (Zn), copper (Cu) and phosphorous (P). High P concentration in the growth medium inhibited55Fe absorption and translocation in both millet and sorghum but millet was more tolerant of P-induced Fe deficiency than sorghum. Millet and sorghum grew best at 1.0 and 0.1 mM P, respectively. Fe concentrations in plant declined as P levels increased; while Mn increased in both millet and sorghum with increasing P levels. Millet, however, maintained higher concentrations of Fe and Mn. The distribution patter of55Fe, regardless of P levels could be summarized as follows: Roots〉Vascular tissue〉leaf-sheaths〉leaf blade. Less55Fe was absorbed and translocated to shoots at 2.5 mM P, in millet and sorghum.
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  • 33
    ISSN: 1432-0878
    Keywords: Auditory nuclei ; Avian brain ; Auditory stimulation ; Deoxyglucose method ; Autoradiography
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The auditory pathway of the Guinea Fowl was labeled with [C14]2-deoxy-D-glucose after stimulation with pure tones, harmonic tones and species-specific calls. In addition to other auditory nuclei, which showed more or less uniform labeling with the present technique, the n. mesencephalicus lateralis dorsalis (MLD) of the midbrain, as well as field L and parts of the hyperstriatum ventrale in the telencephalon, showed a stripe-pattern of labeling after stimulation with a pure tone. The position and orientation of the tone-activated striped areas in field L, observed after stimulation with different tones, correspond to isofrequency contours obtained with microelectrode recordings. The labeling of the three congruent tonotopically organized layers of field L (L1, L2, and L3) was not uniform along the anterior-posterior axis of the field. Harmonic tones produced multiple reactive stripes each of which corresponded to the stripe characteristic of a particular harmonic presented as a pure tone. The species-specific Iambus-call labeled the tonotopic area of field L that corresponds to the frequency band with the highest energy of the call. The hyperstriatum ventrale generally showed a weaker pattern of labeling that, however, resembled the labeling in field L.
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  • 34
    ISSN: 1432-0878
    Keywords: Monoaminergic neurons ; Autoradiography ; Annelid ; Owenia fusiformis (Polychaeta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Light- and electron-microscopic autoradiographic studies were carried out on Owenia to detect selectively catecholaminergic and indolaminergic neurons at two appropriate body levels according to the regenerative properties of this annelid, i.e. in the 1st and the 4th abdominal segments. Autoradiographically, intense 3H-5-hydroxytryptamine accumulation is seen in the 1st abdominal segment, and a less intense autoradiographic reaction in the 4th segment. Roughly, a similar difference in the distribution of the label is seen at both levels following 3H-noradrenaline administration. These observations, confirmed by a quantitative uptake analysis, suggest the presence of catecholaminergic and indolaminergic neurons in Owenia.
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  • 35
    ISSN: 1432-0878
    Keywords: Glucocorticoid ; Antiglucocorticoid ; Mouse brain ; Autoradiography
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Résumé La localisation au niveau du cerveau de souris de la corticostérone, qui est le glucocorticoide naturel chez la souris, et de la cortexolone, démontrée comme étant un antagoniste des glucocorticoides, est étudiée par autoradiographie 30 min après injection in vivo des composés tritiés. Après injection de 3H-corticosterone (3HB), la radioactivité se concentre préférentiellement dans des noyaux cellulaires de plusieurs structures du système limbique et dans les noyaux de certains neurones du cortex cérébral et du bulbe rachidien. Cette concentration nucléaire est abolie après injection de 3H-corticostérone en présence d'un excès de corticostérone non radioactive. Après injection de 3H-cortexolone (3HS), une distribution diffuse de la radioactivité est observée dans tout le cerveau, cependant, une concentration plus élevée de grains d'argent est présente dans la partie ventrale du nucleus arcuatus et dans l'infundibulum. Après injection de 3H-cortexolone en présence d'un excès de cortexolone non radioactive, cette accumulation préférentielle des grains est abolie. L'accumulation de la 3H-cortexolone dans la région hypothalamique suggère que la cortexolone se concentre préférentiellement dans la région cérébrale qui contient les sites de liaison de la dexaméthasone et de plus, les résultats autoradiographiques montrent que le complexe cortexolone-récepteur ne s'accumule pas dans le noyau cellulaire.
    Notes: Summary The localization in the mouse brain of corticosterone, the natural glucocorticoid in the mouse, and cortexolone, reported to be a glucocorticoid antagonist, was studied by autoradiography 30 min after in vivo administration of the tritiated compounds. After 3H-corticosterone (3HB) injection, radioactivity was preferentially concentrated in cell nuclei of several structures within the limbic system, and in nuclei of certain neurones of the cerebral cortex and medulla oblongata. This nuclear concentration was abolished after injection of 3H-corticosterone with an excess of unlabelled corticosterone. After 3H-cortexolone (3HS) injection, a diffuse radioactivity was observed throughout the brain. However, a higher concentration of grains was present in the ventral nucleus arcuatus and in the infundibulum. When excess unlabelled cortexolone was administered with 3H-cortexolone this preferential accumulation of grains was abolished. The accumulation of 3H-cortexolone in the medial basal hypothalamic region suggests that cortexolone concentrates preferentially in dexamethasone (DM) target regions, and in addition the autoradiographic results show that the cortexolone-receptor complex does not accumulate in the cell nucleus.
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    Medical & biological engineering & computing 17 (1979), S. 471-475 
    ISSN: 1741-0444
    Keywords: Bone ; Strain
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    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Specimens of dry human cortical bone of a disc form were subjected to sinusoidal loading under controlled conditions. The results show a nonlinear increase of the measured peak potential with frequency which reaches a limiting value at about 200 Hz. The analysis, which is based on the circuit representation proposed previously by Shamos, indicates that such a behaviour can be attributed to the measuring circuit parameters. It was found that some relevant results can be interpreted qualitatively by similar analysis.
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    Calcified tissue international 26 (1978), S. 47-49 
    ISSN: 1432-0827
    Keywords: Matrix vesicles ; Calcospherites ; Mineralization ; Bone ; Dentin ; Cartilage ; SEM
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Mineral particle cluster, corresponding to calcospherites or matrix vesicle-initiated clusters in calcifying cartilage and dentin and to collagen bundle-related mineralization in lamellar bone, have been isolated from NaOCl solutions used to dissolve the organic matrix in preparation of anorganic mineralizing fronts for scanning electron microscopy.
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    Calcified tissue international 26 (1978), S. 81-84 
    ISSN: 1432-0827
    Keywords: Osteoclasts ; Bone ; Resorption
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    Topics: Biology , Medicine , Physics
    Notes: Summary Osteoclastic bone resorption involves the solubilization of the mineral salts and the degradation of noncollagen bone matrix and collagen fibrils. As no recognizable collagen fibrils have ever been reported within cytoplasmic vacuoles in osteoclasts, it is generally assumed that the collagen fibrils are digested extracellularly in the resorption zone. The extent to which lysis occurs extracellularly and whether or not the osteoclasts phagocytose the degradation products remain to be established. In the present communication, a hypothesis is presented suggesting the possibility that osteoclastic resorption of bone involves the participation of two different cell types. According to this hypothesis, osteoclastic bone resorption is initiated by osteoclasts that demineralize areas of bone and degrade noncollagen bone matrix. After the osteoclasts have moved away or become partially detached from the demineralized site, the exposed collagen fibrils are phagocytosed by mononuclear, fibroblast-like or monocyte-derived cells.
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    Calcified tissue international 26 (1978), S. 155-161 
    ISSN: 1432-0827
    Keywords: Plasma protein ; Glycoprotein ; Bone ; Glucosamine
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    Topics: Biology , Medicine , Physics
    Notes: Summary Plasmaα 2HS-glycoprotein is specifically accumulated in calcified tissues. In the present studies this glycoprotein was isolated from plasma and after iodination with iodine-125 was injected intravenously into young rabbits. The tissue distribution and plasma disappearance rate of this radioactively labeled material were determined. Of the various tissues studied, bone showed the greatest retention of labeled glycoprotein expressed as percentage of the injected dose per gram tissue relative to the plasma content. The rate of loss of iodinatedα 2HS-glycoprotein from plasma was similar to that ofα 2HS-glycoprotein labeled endogenously by using14C-glucosamine or3H-glucosamine. The uptake of exogenously labeled3I-α 2HS-glycoprotein into bone tissue expressed as a percentage of the injected dose was similar to that of endogenously labeled14C-α 2HS-glycoprotein. These results suggest that the125I-labeled material can be used to study further the metabolism ofα 2HS-glycoprotein by bone tissue.
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    Calcified tissue international 26 (1978), S. 209-213 
    ISSN: 1432-0827
    Keywords: Mithramycin ; Bone ; β-Glucuronidase ; KT medium
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    Topics: Biology , Medicine , Physics
    Notes: Summary Mithramycin suppresses bone resorption. Its effect on the synthesis and release of β-glucuronidase (a referent for lysosomal enzymes) in mouse calvarial explants was studied in an in vitro culture system. A newly described medium (designated as KT medium) was introduced in this specific study. Mithramycin initially inhibited the release of β-glucuronidase into the medium and resulted in an ultimate accumulation of this enzyme in the bone. These results suggest that inhibition of bone resorption by mithramycin may be attributed to interference in release of lysosomal enzymes from bone cells.
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    Calcified tissue international 25 (1978), S. 37-43 
    ISSN: 1432-0827
    Keywords: ESR ; Radiation free radicals ; CO3-Apatites ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Exposure of bone mineral to X-rays generates free radicals. These are usually very labile, but can be stabilized at liquid nitrogen temperatures for study by electron spin resonance spectroscopy. The free radicals thus detected in the present study included one with resonances arising from an electron excess center and 2 species with electron-deficit centers: a phosphate anion radical and a radical associated with carbonate. Each of these radicals seemed to be located chiefly at the mineral surface and was sensitive to the surface environment. Presence of an organic phase, as in whole bone, modified free radical production in a manner that suggests interference with the formation of electron deficit centers. Comparison with other synthetic minerals suggests that precipitated carbonateapatites are good models for bone mineral.
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    Calcified tissue international 25 (1978), S. 227-232 
    ISSN: 1432-0827
    Keywords: Parathyroid hormone ; Bone ; Calcium ; Chick embryo ; Organ culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The purpose of this study was to measure in an in vitro system the movement of Ca and phosphate (Pi) out of bone when treated with parathyroid hormone (PTH). Tibiae from 13-day chick embryos were incubated for up to 8 h in a defined medium containing 1.8 mM Ca. Medium samples were collected every 2 h and were analyzed for Ca, Pi and lactate. Net effluxes from the bones were calculated. When bones were incubated with PTH in the medium (1 U/ml), net Ca efflux was increased 44, 60 and 100% at 4, 6 and 8 h, respectively. At no time was net Pi efflux affected by the hormone. The well known PTH-stimulated lactate production was not seen until 8 h. Lower doses of PTH (0.1 and 0.3 U/ml) were also effective. Comparing PTH (1 U/ml) responsiveness at higher (2.2 mM) and lower (0.9 mM) medium Ca concentrations, showed that with 2.2 mM Ca no increased Ca efflux was seen, while with 0.9 mM Ca significant elevation in medium Ca occurred 2 h sooner than in the experiments using 1.8 mM Ca. In another experiment, varying the medium Pi level from 1 to 2 mM had no effect on the Ca response to PTH. In neither experiment was Pi release affected by PTH. The results of this study have led to the following conclusions: (1) PTH acts on bone to cause an early dose related increase in net Ca efflux; (2) the effect is specific for Ca, since it is not accompanied by an increased Pi efflux, and may be saturated by raising the medium Ca level; and (3) PTH-stimulated Ca efflux in this system is not correlated with, and is probably not a result of increased lactate production.
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  • 43
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    Calcified tissue international 25 (1978), S. 233-240 
    ISSN: 1432-0827
    Keywords: Bone ; Bone resorption ; Albumin
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    Topics: Biology , Medicine , Physics
    Notes: Summary A fraction (brA), which causes resorption of fetal rat bones in vitro, has been concentrated from bovine serum albumin by anion exchange column chromatography on DEAE Sephadex. This active fraction has also been prepared using DEAE Sephadex A-50 by a batch method with a 0.09M NaCl, 0.1M TRIS buffer, pH 8.35. BrA was 10–30 times more potent than the original albumin. The retained material, which constitutes the bulk of the protein and has less activity than the original albumin, elutes with 0.45M NaCl. Similar treatment of serumα,β or γ globulins does not yield brA. Further enhancement of the bone resorbing activity of brA can be obtained with (NH4)2SO4 fractionation or extraction with CH3OH∶CHCl3. Heating at 55° C for 2 h or at 100° C for 10 min does not affect the activity; overnight incubation with protease destroys the bone resorbing effect. The bone resorbing activity is not removed by dialysis and does not correlate with the protease activity of the fraction. The action of brA is inhibited by 3 mM PO4, 1 μg/ml calcitonin or glucagon, 10−7 M dexamethasone or 0.02 μg/ml actinomycin D. The bone resorbing activity of brA is partially inhibited by 10−7–10−5 M indomethacin. PTH did not elicit bone resorption when added to cultures incubated in chemically defined medium supplemented with 0.1 mg/ml brA. However, brA did not inhibit PTH-induced resorption.
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  • 44
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    Calcified tissue international 25 (1978), S. 249-254 
    ISSN: 1432-0827
    Keywords: Bone ; Oestrogens ; Oestrogen receptors ; Binding proteins
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    Topics: Biology , Medicine , Physics
    Notes: Summary Attempts were made to demonstrate the presence of specific oestrogen binding proteins (“receptors”) in bone cells. High speed cytosol preparations of bone were incubated with several concentrations of radioactive oestradiol alone and with radioactive oestradiol in the presence of a specific antioestrogen, Nafoxidine. Separation of bound and free oestradiol was carried out by dextran coated charcoal treatment and by sucrose gradient ultracentrifugation. Several types of bones likely to be oestrogen-sensitive were investigated: human femoral heads, human phalanx, rat and rabbit calvaria, humeri and femora of female rats. In all experiments we were unable to demonstrate the presence of specific oestrogen receptors in bone cell cytosol indicating that the direct effect of oestrogens on bone, if present, is not mediated by specific oestrogen receptors.
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  • 45
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    Calcified tissue international 26 (1978), S. 85-89 
    ISSN: 1432-0827
    Keywords: Bone ; Histology ; Microradiography ; Densitometry ; Computer
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    Topics: Biology , Medicine , Physics
    Notes: Summary A comparatively simple quantitative method for assessing bone morphology has been evolved. Microradiographs of thin sections of mandible have been scanned with a Joyce-Loebl double beam recording microdensitometer with a scanning autodensidater attachment, using a white beam. For each image the optical densities for all the pixels (picture elements) were divided into 10 groups. The limits of the division were fixed by the maximum and minimum densities occurring within the image. A computer generated map was produced which indicated the spatial distribution of the pixels within each group to which an arbitrarily chosen shading was attached. The number of pixels within each group is also shown on the map. The computer map was compared with the photomicrograph and, where necessary, the original section. The fractional area of hard tissue was then readily determined using the numerical values of each group of pixels.
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  • 46
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    Calcified tissue international 26 (1978), S. 139-142 
    ISSN: 1432-0827
    Keywords: Chemisorption ; Surface area ; Hydroxyapatite ; Bone ; Enamel
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    Topics: Biology , Medicine , Physics
    Notes: Summary The surface areas of three different samples of hydroxyapatite, samples of deproteinized bone, and samples of whole and deproteinized enamel were determined by adsorption of an adduct (the diglycidyl ether of bisphenol A with N-phenylglycine) from methylene chloride solution. In all cases, the surface areas of these samples agree well with those obtained by the BET (N2) method.
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  • 47
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    Calcified tissue international 25 (1978), S. 105-110 
    ISSN: 1432-0827
    Keywords: Progesterone ; 17β estradiol ; Testosterone ; Tissue culture ; Bone
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    Topics: Biology , Medicine , Physics
    Notes: Summary Although sex steroids are known to affect skeletal metabolism, their effects on bone collagen synthesis have not been studied. We have examined the direct effects of progesterone, 17β estradiol, testosterone, 5α dihydrotestosterone and dehydroepiandrosterone on bone collagen and noncollagen protein synthesis in cultures of calvaria obtained from 21-day fetal rats. Bones were incubated for 24 to 168 h and3H-proline was added for the last 2 h of culture. Incorporation of the label into collagenase-digestible protein (CDP)1 and noncollagen protein (NCP) was determined using repurified bacterial collagenase. Progesterone caused a dose dependent inhibition of the labeling of CDP at concentrations of 3×10−7 M to 10−5 M after 96 h of culture. A smaller inhibitory effect was observed on NCP. The inhibitory effect was slow in onset and persisted when bones were incubated for 48 h with progesterone and then transferred to control medium for 48 h. Progesterone also inhibited the incorporation of3H-thymidine and3H-uridine into fetal rat calvaria. After 24 h of culture, 17β estradiol and testosterone had no effects on the labeling of CDP and NCP. After 96 h, 17β estradiol had a small and inconsistent stimulatory effect on the labeling of CDP but testosterone had no effect. Neither hormone altered the inhibitory effects of parathyroid hormone, cortisol and progesterone. Dihydrotestosterone and dehydroepiandrosterone had no effects after 24 h and 96 h of culture. 17β estradiol, testosterone and dihydrotestosterone did not affect the incorporation of3H-uridine or3H-thymidine into fetal rat calvaria. Our studies indicate that progesterone is an inhibitor of bone collagen synthesis and estrogens and androgens are not major regulators of bone formation in vitro.
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  • 48
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    Calcified tissue international 25 (1978), S. 29-35 
    ISSN: 1432-0827
    Keywords: Albumin ; Bone ; Growth retardation ; Osteomalacia
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    Topics: Biology , Medicine , Physics
    Notes: Summary 125I-labeled rat albumin injected intravenously into rats was taken up by growing bone. Some of this radioactive albumin could be removed from bone by washing with saline, the proportion so removed decreasing from 82.5% at 1 day to 7.4% at 8 days. Both the total radioactivity, and that remaining in bone after saline wash, were reduced when growth was slowed by alteration or restriction of the diet. Although the amount of125I albumin in rachitic bone was reduced, autoradiography showed that radioactivity was present in rachitic osteoid. Immunoprecipitation using anti-rat-albumin serum showed that about half of the radioactivity released from normal bone by EDTA was still attached to albumin. These results suggest that albumin plays some part in the growth of bone.
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  • 49
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    Archives of microbiology 118 (1978), S. 27-34 
    ISSN: 1432-072X
    Keywords: Bacillus subtilis ; Sporulation ; Uracil incorporation ; Autoradiography ; Nuclear shape
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    Topics: Biology
    Notes: Abstract The transcriptional activity of the two genomes of the sporangium during spore formation was determined by pulse-labeling bacteria with 3H-uracil at different times of sporulation and preparing them for high resolution autoradiography. The quantitative analysis of autoradiographs shows that uracile incorporation in the whole sporangium decreases considerably between stages II and IV. However, the variations of the transcriptional activity are not identical in the mother cell and in the forespore. The one of the mother cell decreases rapidly between stages II and III and then remains stable until the end of stage IV, whereas that of the forespore which is low at stage II increases as the forespore grows ovoid and then quickly diminishes. It is very weak at the beginning of stage IV and negligible at the end of this stage. Pulse-chase experiments made in the presence of rifampine indicate that about 30% of the uracile incorporated is located in stable RNA. This value is found at any stage of sporulation in both cellular compartments whatever their rate of uracile incorporation. A relationship can be made between the nuclear shape and the activity of the genetic material. This confirms observations made by several authors in other bacterial species and other physiological conditions that the condensed shape corresponds to a state of low transcriptional activity whereas the more irregular and dispersed shape corresponds to a state of high activity.
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  • 50
    ISSN: 1432-0878
    Keywords: Suboesophageal ganglion (cricket) ; Paraamedial neurosecretory cells ; Stainability ; Incorporation of 35S-cysteine ; Autoradiography
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    Topics: Biology , Medicine
    Notes: Summary Ovariectomy, performed immediately after the final hatch, caused a reduction of stainable (neurosecretory?) material in the paramedial neurosecretory cells (PNC) (A-type) of the suboesophageal ganglion in 10 day-old females of Teleogryllus commodus (Walk.). A concomitant increase in nuclear volume and in the incorporation of 35S-cysteine indicates increased synthesis of neurosecretory material. From these findings it is concluded that more stainable material is secreted in the cerebral neurohaemal organ after Ovariectomy. A functional relationship between the PNC and the ovaries is suggested.
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  • 51
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    Cell & tissue research 186 (1978), S. 535-540 
    ISSN: 1432-0878
    Keywords: Skeletal muscle ; Satellite cells ; Regeneration ; Electron microscopy ; Autoradiography
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    Topics: Biology , Medicine
    Notes: Summary Satellite cells were traced autoradiographically during the regeneration of skeletal muscle in young Sprague-Dawley rats. Approximately 31% of the satellite cells in uninjured muscles appeared labelled after three injections of tritiated thymidine; none of the myonuclei were labelled in the same muscles. Four to six days after transplanting the radioactive muscles to non-radioactive littermates, regenerating myotube nuclei in the host appeared labelled. Thus, this study confirms that satellite cells in young rats can differentiate into multinucleated myotubes following muscle injury.
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  • 52
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    Cell & tissue research 189 (1978), S. 305-312 
    ISSN: 1432-0878
    Keywords: Adrenal medullary cells (mouse) ; Proliferative activity ; Autoradiography ; Formaldehydeinduced fluorescence
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    Topics: Biology , Medicine
    Notes: Summary The proliferative activity of newborn and adult mouse adrenal medullary cells was determined with light and electron microscopic autoradiography. The H3 thymidine labelling index of 2 weeks old mice adrenal medullary cells was about 9.4 % and declined to less than 1 % in adult mice. In electron microscopic autoradiography labelled norepinephrine as well as epinephrine cells could be seen. Only in 1 and 2 weeks old mice some morphologically undifferentiated cells were visible. In formaldehyde induced fluorescence combined with light microscopic autoradiography the fluorescence intensities of labelled and unlabelled medullary cells were measured. On average the fluorescence intensity of labelled cells was lower than that of unlabelled cells. The differences could be explained by a higher number of autoradiographic silver grains laying on the cytoplasm of labelled cells. These results give evidence that fully differentiated adrenal medullary cells are capable of division.
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  • 53
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    Cell & tissue research 193 (1978), S. 259-267 
    ISSN: 1432-0878
    Keywords: Skeletal muscle ; Denervation ; Tenotomy ; Cell proliferation ; Autoradiography
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    Topics: Biology , Medicine
    Notes: Summary Autoradiographic experiments using 3H-thymidine were designed to analyse cell proliferation which occurs in skeletal muscle after denervation and after tenotomy. In mouse tibialis anterior and tongue muscles during the first 24 h after denervation or tenotomy labelling levels were low and did not differ significantly from sham operated control muscles. By 48 h after denervation and tenotomy of tibialis anterior muscles, increased levels of labelling occurred in both muscle and connective tissue nuclei. Daily pulse labelling for 7 days after denervation produced a labelling level which was 8 times that of sham operated controls, 25–30% of the total nuclear population being labelled. Denervated muscles had twice the level of labelling compared to tenotomised muscles. These results provide conclusive evidence that both denervation and tenotomy stimulate cell proliferation in skeletal muscle and it is suggested that the increased numbers of labelled muscle nuclei are likely to be the result of mitotic activity in muscle satellite cells.
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  • 54
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    Cell & tissue research 194 (1978), S. 279-285 
    ISSN: 1432-0878
    Keywords: 14C-thymidine incorporation ; Autoradiography ; Multinucleated blasts ; Electron microscopy ; Deformities of cellular membranes
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    Topics: Biology , Medicine
    Notes: Summary Cytochalasin B (CB) administered simultaneously with a mitogenic dose of concanavalin A (Con A) interferes with the activation process. This interference involves structural alterations of cellular membrane which do not include a reduced Con A-binding capacity. This conclusion is supported by the observation of deformities in both nuclear and cytoplasmic membranes in Con A-activated lymphocytes subsequently treated with CB. The high incidence of membrane blebs and pseudomyelin bodies in the cytoplasm points to a general effect of CB on the structural organization of membrane which may secondarily interfere with some specific event such as generation or transfer of signals for activation or cytokinesis.
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  • 55
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    Cell & tissue research 195 (1978), S. 29-43 
    ISSN: 1432-0878
    Keywords: Endocrine hypothalamus (rat) ; Arcuate nucleus ; L-dopa ; Autoradiography ; Electron microscopy
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    Topics: Biology , Medicine
    Notes: Summary Light and electron microscopic autoradiography has been employed to define the neuroanatomical patterns of uptake and binding of radiolabelled L-dopa in the endocrine hypothalamus of the rat. A dorsomedial continuum of arcuate and periventricular neurons selectively sequester 3H L-dopa 20 min following its intraventricular infusion. By 40 and 60 min following the infusion labelling of neurons is minimal and supports the notion of rapid degradation. Other cell compartments such as tanycytes demonstrate uptake of 3H L-dopa. The ultrastructural localization and distribution of radiolabelled L-dopa (or its metabolites) in the rodent hypothalamus is discussed with respect to mechanisms and cell compartments involved in neuroendocrine regulatory processes.
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  • 56
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    Cell & tissue research 191 (1978), S. 183-186 
    ISSN: 1432-0878
    Keywords: Amygdaloid body ; Hypothalamus ; Midbrain ; Autoradiography ; Rat
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    Topics: Biology , Medicine
    Notes: Summary Light microscopic autoradiography was performed subsequent to injection of tritiated amino acids into various parts of the amygdaloid body of the rat. Evidence is provided for two hitherto unreported projections of the amygdala: from the medial amygdaloid nucleus to the contralateral premamillary nuclei and from the central amygdaloid nucleus to the mesencephalic central grey. The functional implications of these findings are discussed.
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  • 57
    ISSN: 1432-0878
    Keywords: Bone marrow (Rat), Stroma ; Regeneration and hemopoiesis after evacuation ; Morphometry ; Histogenesis ; Autoradiography
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    Topics: Biology , Medicine
    Notes: Summary The marrow cavity of the rat tibia was mechanically evacuated and autoimplanted to the subcutaneous tissue. The regenerative process which restored the integrity of marrow stroma and hemopoiesis, was morphometrically evaluated in whole mount of tibia. Following evacuation, the clot filled the cavity. The granulation tissue then appeared and expanded, penetrating and replacing the clot. The fibroblasts of the granulation tissue differentiated into osteoblasts forming osteoid bone. Within its interstices, the primordial marrow consisting of loose connective tissue and vascular sinuses appeared and hemopoiesis resumed. Expansion of hemopoiesis resulted in the resorption of bone and within three weeks the tibial cavity was restored to the pre-evacuation state. Autoradiography indicated that the labeling index was initially high in fibroblasts and osteoblasts but was subsequently reduced while it increased in osteocytes, cells of Haversian canals, stromal and hemopoietic cells of marrow. The finding is in disagreement with the view that the regenerative process originates from the Haversian canal. When the label was introduced on day 4 post-operatively, it subsequently appeared in osteocytes, cells of Haversian canal, stromal elements of the marrow, but not in the hemopoietic cells. This indicates complete dissociation of marrow stroma and hemopoietic stem cell.
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  • 58
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    Cell & tissue research 194 (1978), S. 103-114 
    ISSN: 1432-0878
    Keywords: Polyamines ; α-methylornithine ; Gastrulation ; Autoradiography ; Polychete
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    Topics: Biology , Medicine
    Notes: Summary Continuous treatment of developing eggs of the polychete Ophryotrocha labronica with α-methylornithine, which inhibits synthesis of putrescine, led to arrest of development at gastrulation. The present ultrastructural analysis suggests that the arrest of development is due to failure to form nuclei, and thus reveals a possible role for putrescine in nucleolar formation. Further support for this contention was provided by means of electron-microscopical autoradiography. It was found that newly synthesized putrescine, derived from administered 3H-ornithine, labeled the nucleoli intensely at the time of their normal appearance during gastrulation, the time at which the rate of endogenous putrescine synthesis is maximal. These observations have led to the conclusion that putrescine synthesis may be directly involved in formation of nucleoli.
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  • 59
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    Cell & tissue research 194 (1978), S. 207-218 
    ISSN: 1432-0878
    Keywords: Protein transport ; Kidney tubules (proximal) ; Muramidase (lysozyme) ; Autoradiography ; Hypothetical grain analysis
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    Topics: Biology , Medicine
    Notes: Summary Proteins filtered in the renal glomeruli are reabsorbed by the proximal tubule and catabolized in the lysosomes. On the basis of studies on isolated flounder tubules it has been suggested that, in addition to this catabolism, a transtubular transport of intact protein (lysozyme) also occurs. The present study demonstrates that significant amounts of lysozyme are reversibly bound to the peritubular side of isolated tubules. Electron microscopic autoradiography demonstrates that the protein is located in the basement membrane and intercellular spaces. After in vivo injection, 125-Ilysozyme is mainly located in endocytic vacuoles of the first proximal segment, but also over the basal part of the cells. Since a significant peritubular binding of lysozyme is demonstrated in vitro, it is suggested that a similar binding of tracer protein originating from the peritubular capillaries might occur in vivo and that subsequent release of this protein in vitro might simulate transtubular transport. It is therefore concluded that release of tracer protein from isolated kidney tubules does not conclusively demonstrate transtubular transport of intact protein in experimental systems in which peritubular binding of protein can be demonstrated.
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  • 60
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    Calcified tissue international 22 (1977), S. 303-313 
    ISSN: 1432-0827
    Keywords: Osteoclasts ; Sedimentation ; Isolation ; Cells ; Bone
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    Topics: Biology , Medicine , Physics
    Notes: Abstract A method is presented for separating osteoclasts from the heterogeneous population of bone and marrow cells. Cell suspensions were prepared from femora of young rabbits by mechanical dispersion. The starting cell suspension typically contained only 1.0%±0.5 osteoclasts. Following an initial 45 min of unit gravity sedimentation in a lucite chamber osteoclasts were primarily distributed in fractions 2–5. A second 45-min sedimentation of these pooled fractions yielded cell suspensions containing greater than 30% osteoclasts (as much as a 50-fold increase over starting percentages). Linear scan analysis, however, revealed that osteoclasts accounted for 73.14%±0.58 of the cell colume. Subsequent in vitro experiments demonstrated linear incorporation of3H-leucine into TCA precipitable protein for cells comprising the osteoclast fraction. Concomitant radiautographs revealed radioactive label in isolated osteoclasts.
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  • 61
    ISSN: 1432-0827
    Keywords: Bone ; Diphosphonates ; Bone resorption ; Bone formation
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    Notes: Summary Male rats weighing 100 g were assigned to groups and injected daily for 10 days with vehicle (control), 0.4, 2.0, 4.0, 10.0, or 20.0 mg/kg/day of ethane-1-hydroxy-1,1-diphosphonate (EHDP) or dichloromethylene diphosphonate (Cl2MDP). The proximal tibial metaphysis and epiphysis were assayed for changes in percentage of hard tissue and bone formation parameters. From the data, information about hard tissue resorption was deduced. All doses of Cl2MDP and doses of 2.0 mg EHDP/kg/day and greater caused significant increases in percentage of hard tissues with Cl2MDP being more effective than similar doses of EHDP in decreasing bone resorption. Osteoclast population parameters were increased with all doses of both Cl2MDP and EHDP with Cl2MDP having a greater effect than similar doses of EHDP. Decreases in the proliferation of the osteoprogenitor pool parallel the decreases in osteoblasts and bone formation parameters. These decreases in osteoprogenitor pool proliferation do not account for the increases with diphosphonates in osteoclast population parameters.
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  • 62
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    Calcified tissue international 23 (1977), S. 283-289 
    ISSN: 1432-0827
    Keywords: Bone ; Resorption ; Osteoclast ; Quantitation
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    Notes: Summary The stimulation of bone resorption, assessed by the release of45Ca from prelabeled bones, was associated with an increase in number of osteoclasts per bone section in parathyroid hormone (PTH)-treated bones, but not in lipopolysaccharide (LPS)-treated bones. By contrast the number of nuclei per osteoclast increased following LPS treatment, but was not affected by PTH. LPS-treated bones had more multinucleated cells, some having as many as 27 nuclei per osteoclast. More osteoclasts were adjacent to the bone collar in bones treated with LPS or PTH than in control bones. In LPS-treated bones this area also contained the largest osteoclasts, as determined by the greatest number of nuclei per osteoclast. The results suggest that LPS and PTH stimulate osteoclastic resorption by different mechanisms.
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  • 63
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    Calcified tissue international 23 (1977), S. 303-305 
    ISSN: 1432-0827
    Keywords: Albumin ; Bone ; Hydroxyapatite ; Plasma protein
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    Notes: Summary The extractability of125I-labelled plasma albumin from bone pieces and from powdered bone has been compared after both in vivo and in vitro incorporation. The results show that albumin is more readily extracted from bone pieces than from bone powder which implies that tissue disruption exposes additional protein adsorption sites. It is suggested that incorporation of plasma albumin into calcified matrix during bone formation occurs mainly as a result of its strong interaction with bone mineral.
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  • 64
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    Calcified tissue international 23 (1977), S. 161-170 
    ISSN: 1432-0827
    Keywords: Bone ; Growth ; Rat ; Remodeling ; Tetracyclines
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    Topics: Biology , Medicine , Physics
    Notes: Summary Cortical bone remodeling along the femur diaphysis was determined in normal female rats (Sprague-Dawley) with the tetracycline technique. Three segments on the cortical bone circumference (the anterolateral, the medial, and the posterior) were found to be most suitable for the study of the remodeling process. Oxytetracycline was administered at age 60 and 75 days, and groups of animals were killed at age 75, 85, 95, and 105 days. The accumulated endosteal growth during age 60 to 75 days in the anterolateral segment was found to increase uniformly in the distal direction along the femur diaphysis. A method is described where this accumulated endosteal growth is used. This method eliminates the use of calipers to determine the section level and makes it possible to study comparable sections even after varying periods of time. The proximal part of the diaphysis showed periosteal apposition in all three segments. The periosteal apposition turned into resorption in the distal part of the diaphysis in the anterolateral and medial segments, whereas the periosteal appsition increased in the posterior segment. The endosteal growth increased in the distal direction in the anterolateral and medial segments. Irregular OTC bands made measurements of endosteal remodeling in the posterior segment impossible. The cortical width decreased in the distal direction along the femoral shaft. Comparison between the different age groups is described and also the relation between the accumulated endosteal growth and the diameter of the medullary cavity.
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  • 65
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    Calcified tissue international 23 (1977), S. 1-5 
    ISSN: 1432-0827
    Keywords: Water ; Bone ; Homeostasis ; Structure
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    Notes: Summary A short review is given of the water in bone. Various analyses of bone water content are discussed, and its possible location is considered in relation to the behaviour of water in isolated components of bone. Some of the difficulties encountered in examining such microscopic phenomena as water structure in a heterogeneous system such as bone are also discussed.
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  • 66
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    Calcified tissue international 24 (1977), S. 41-46 
    ISSN: 1432-0827
    Keywords: Lead salt ; Bone ; Mobilization ; Hypercalcemia ; Hyperphosphatemia
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    Topics: Biology , Medicine , Physics
    Notes: Summary The present study is an investigation of the mechanism of hypercalcemia and hyperphosphatemia induced by the intravenous injection of lead acetate (Pb-Ac). A total of 118 male rats were injected with 30 mg/kg of Pb-Ac, or with 16.5 mg/kg of sodium acetate as the control. The levels of serum calcium, phosphorus and lead were then determined at various time periods after the injections. Serum calcium and phosphorus levels increased with time after Pb-Ac injection and the maximum values of calcium (17 mg%) were found after 1 h and of phosphorus (13.5 mg%) after 30 min. Both calcium and phosphorus levels reverted to the normal range after 12 h. The maximum net rates of increase of calcium and phosphorus were found immediately after Pb-Ac injection. At that time, deposition of lead at the calcifying sites of bone and incisor dentin was demonstrated by a histochemical examination. In other experiments the changes in the calcium and phosphorus contents in the medium after shaking bone powder in serum with Pb-Ac in an in vitro system were studied. It was confirmed that the calcium and phosphorus were displaced from the bone mineral, the extent of the displacement being correlated with the concentration of the Pb-Ac added to the medium, and that these displacements were very rapid reactions. These results suggest that hypercalcemia and hyperphosphatemia following Pb-Ac injection results from a direct action of lead on the bone mineral.
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    Calcified tissue international 24 (1977), S. 209-214 
    ISSN: 1432-0827
    Keywords: Parathyroid hormone ; Plasma calcium ; Radiocalcium ; Bone
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    Topics: Biology , Medicine , Physics
    Notes: Summary The net rate of loss of45Ca from plasma was monitored in normal, thyroparathyroidectomized (TPTX), and nephrectomized rats. Also studied was the effect of a 24 h intravenous infusion of PTH in TPTX rats. The45Ca was injected after an overnight fast and 1 to 4 h prior to the time the first blood sample was obtained. In normal rats, the net rate of loss of45Ca from plasma was slower than in TPTX rats for at least 12 h after radionuclide injection. Nephrectomy did not affect this difference between control and TPTX rats until 8 h after45Ca injection or about 18 h after nephrectomy. At this time a reversal occurred and the net rate of plasma45Ca loss in nephrectomized rats with parathyroids became faster than in TPTX nephrectomized rats. When PTH was infused at a constant rate (3 mU/g body weight/h) plasma calcium concentrations rose gradually for approximately 12 h after which a constant plasma calcium level was maintained with continued PTH infusion. During the period when plasma calcium concentrations were rising, the net rate of loss of45Ca from plasma was slower than in TPTX controls infused only with saline. However, when plasma levels stabilized, plasma45Ca disappearance rates changed such that the rate loss of45Ca from plasma became faster in PTH-infused than in TPTX controls. It is concluded that PTH increases calcium efflux from the same compartment in bone which receives calcium from plasma, thereby returning some recently deposited45Ca to plasma. In the non-equilibrated state, this returns additional45Ca to plasma causing anet decrease in the rate of loss of45Ca from plasma. However, in the equilibrated condition, when both calcium efflux from bone and influx into bone are in balance but at higher flux rates, PTH stimulates the diffusion of45Ca into other bone compartments.
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  • 68
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    Calcified tissue international 23 (1977), S. 185-188 
    ISSN: 1432-0827
    Keywords: Matrix vesicles ; Bone ; Ultracrytomy
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    Topics: Biology , Medicine , Physics
    Notes: Summary Matrix vesicles were observed in femurs of 8-day-old chick embryos prepared by ultracryotomy. Some of the sections were subjected to ultramicroincineration. The unfixed tissues never came into contact with solutions, and thereby artifacts due to dissolution, redistribution, or rearrangement of the mineral constituents were avoided. In the osteoid, electron dense objects with the size and appearance of matrix vesicles were seen, although limiting membranes were not visible. After ultramicroincineration the vesicles were observed to contain small crystals and a less dense amorphous mineral material which may be the precursor of bone mineral. In addition, a ring of ash enclosed the crystalline and amorphous mineral and appeared to occupy the position of the vesicle membrane as seen in conventionally prepared material.
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  • 69
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    Calcified tissue international 24 (1977), S. 93-98 
    ISSN: 1432-0827
    Keywords: Magnesium ; Bone ; Mineral ; Chick embryo ; Organ culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Using bone organ culture techniques, three concentrations of magnesium were evaluated for their effects on the mineral mass of embryonic chick tibiae incubated for 3 days with or without a metabolic inhibitor (1 mM iodoacetic acid) added to the media. Varying the medium Mg level from 0.3 to 3.0 mM had little effect on the net increase in mineral in live bones (without the inhibitor), but in dead tibiae (with the inhibitor) there was a marked net decrease in mineral deposition with increasing Mg concentration. Bone Mg mass varied directly with the level of Mg in the medium regardless of whether or not the tibiae were inhibited. Responsiveness to parathyroid hormone (PTH) with regard to mineral loss and increased lactate production was not affected by Mg even in tibiae incubated in Mg-free media. However, PTH-treated bones cultured in Mg-free media had a significantly elevated Mg mass, suggesting that the hormone may have a Mg-conserving effect on bone during severe Mg deficiency. The findings of this study support the hypothesis that bone cells regulate the level of Mg to which newly forming mineral is exposed and that PTH-responsiveness in embryonic chick tibiae is not dependent on the ambient Mg concentration.
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  • 70
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    Calcified tissue international 22 (1977), S. 9-17 
    ISSN: 1432-0827
    Keywords: Bone ; Fluoride ; Calcium ; Phosphorus ; Mechanical Strength
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    Topics: Biology , Medicine , Physics
    Notes: Abstract Four groups of weanling rats were fed for 2 weeks on a diet sufficient or insufficient in calcium and/or phosphorus. Each group was divided into four subgroups which were offered distilled water supplemented with 0, 50, 75, or 150 ppm fluoride. High levels of fluoride in drinking water inhibited weight gain. This inhibition was less in rats deficient in phosphorus than when normal-phosphorus diets were offered. At a low level, fluoride was without any effect on bone ash, thickness of femoral cortical bone, and mechanical strength, as measured by maximal load, ultimate stress to breaking, and limit of elasticity. Modulus of elasticity was decreased. At higher levels fluoride tended to decrease most of these parameters, except in rats deprived of both calcium and phosphorus. The effect of fluoride was modified by lack of dietary calcium and/or phosphorus and appeared to be weaker in rats deficient in these nutrients. Lack of dietary calcium and/or phosphorus decreased bone strength more than did fluoride content of water and of bone mineral. Concentration of bone ash and thickness of femoral cortical bone were closely correlated with parameters of mechanical strength.
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  • 71
    ISSN: 1432-0827
    Keywords: Parathyroid hormone ; Plasma calcium ; Radiocalcium ; Bone
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    Topics: Biology , Medicine , Physics
    Notes: Abstract Acute changes in plasma calcium and45Ca were studied in young adult male thyroparathyroidectomized (TPTX) rats injected with moderate doses of parathyroid hormone (PTH). For plasma calcium changes, comparison was made between rats fasted or fed prior to PTH injection. For plasma45Ca changes, the effect of the time of administration of the radionuclide was also studied; this included rats injected with PTH 1 h after radionuclide (“1 h45Ca”), 18 h later (“18 h45Ca”) and more than 6 days later (“6 day45Ca”). The results can be summarized as follows: (1) Plasma calcium changes were greater when PTH was injected into “fed” rather than into “fasted” rats. (2) PTH always produced a relative increase (compared to controls tested concurrently) in plasma45Ca concentrations. This increase was the same in the “1 h45Ca” and the “18 h45Ca” groups. (3) Plasma45Ca rose at least temporarily following PTH injection in the “18 h45Ca” group. (4) The45Ca rise following PTH was always greater in “fed” than in “fasted” groups. (5) Plasma45Ca specific activities (S.A.) tended to rise in the “6 day45Ca” group and to fall in the “18 h45Ca” group, following PTH injection. However, the45Ca S.A. was always higher in fed than fasted groups. (6) In a few experiments in which32P was injected with45Ca, specific activity changes in plasma45Ca following PTH injection werenot accompanied by similar changes in32P specific activity. These results could not be adequately explained by PTH effects on bone resorption, but the data supported the postulate that PTH controls plasma calcium concentrations by increasing transport of calcium through the osteocyte-lining cell (osteoblast) bone cell complex from the bone fluid compartment to the ECF.
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  • 72
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    Calcified tissue international 22 (1977), S. 275-284 
    ISSN: 1432-0827
    Keywords: Bone ; Calcium ; Collagen ; Dentine ; Phosphoprotein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Calcium-binding and biochemical studies have been applied to characterize the state of the carboxylate and protein-bound phosphate groups in the EDTA-demineralized matrices of rat bone and dentine. The organic phosphate and carboxylate content of demineralized bone is virtually identical to that of purified steer skin collagen whereas demineralized dentine has a significantly higher phosphate and carboxylate content, presumably due to the presence of an acidic non-collagenous phosphoprotein. Two classes of calcium-binding sites can be detected in demineralized bone, demineralized dentine, and purified, reconstituted collagen. The number of strong calcium-binding sites correlates with the number of protein-bound phosphate groups. Depending on the preparative procedure, seven to nine such sites (per collagen molecule) are present in dentine, and one to two in the purified reconstituted collagen and in bone. The binding constant for the dentinal sites (1.1×104 M−1), however, is 20 times greater than that for bone or reconstituted collagen fibrils from skin. We tentatively conclude that the strong calcium-binding site in bone and reconstituted collagen is of the form protein-PO 4 − Ca++ whereas in dentine it is of the form the weak binding sites in bone and dentine are of the form protein-COO-Ca++; and that approximately 160 of the 217 carboxylate groups of the collagen molecules of dentine or bone are present as electrostatic linkages of the form protein-COO−+H3N-protein.
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  • 73
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    Calcified tissue international 22 (1977), S. 315-327 
    ISSN: 1432-0827
    Keywords: Bone ; Bioelectricity ; Strain ; Locomotion ; Osteoporosis
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    Topics: Biology , Medicine , Physics
    Notes: Abstract Using wet dead specimens it was possible to show that the size and form of strain-related electrical potentials on the surface of sheep radii were related both to the amount of strain, and the strain rate, over the range of these recorded during locomotion. Using the same electrode and amplifier system in vivo changes in surface strain and surface charge were recorded from the radius of three sheep during locomotion. During slow locomotion electrical changes were negligible. At medium speed they were most variable, and profoundly influenced by small alterations in the timing and pattern of strain change. When locomotion was fast the electrical waveform reflected fairly faithfully the changes in strain on the bone surface.
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  • 74
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    Calcified tissue international 22 (1977), S. 329-331 
    ISSN: 1432-0827
    Keywords: Bone ; Fracture ; Mineral
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    Notes: Abstract The bone mineral content was measured by gamma absorptiometry in patients who had sustained fractures of the upper limb. Measurements were taken on the shafts of the forearm and in the trabecular bone immediately proximal to the wrist. Fractures of the surgical neck of the humerus and fractures of the shaft of the humerus did not significantly influence the bone mineral content of the ipsilateral forearm. Fracture of the radius and the ulna, however, caused a 15% loss of the bone mineral content in the trabecular bone proximal to the wrist, whereas the mineral content on the shafts of the fractured forearms increased or decreased depending on the fracture site. The findings suggest that measurements of bone mineral content of the forearm are not necessarily invalidated if there has been a fracture of the same limb. Only if the forearm itself is fractured should measurements on this site be avoided.
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  • 75
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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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  • 76
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    Keywords: Bone ; Calcium ; Strontium ; Apposition ; Tracer analysis
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    Notes: Abstract 1. Evidence has accumulated that the rate of acceretion (A) of calcium to bone is the sum of two fluxes; apposition involving the laying down of new bone and augmentation which is the result of slow exchange of non-surface bone calcium with plasma calcium pools as the result of solid state diffusion. 2. A method has been devised for separating A into its two components. It requires the use of45Ca or, for clinical studies,85Sr as a calcium tracer. Studies which are initiated with a combined accretion rate-calcium balance study, are concluded with an estimate of the exponent of the power function which has been found to describe the whole body retention of tracer from the second month onward. 3. The impulse response function of the skeleton for the tracer is then calculated, making the assumption that in any uniform volume of bone, osteoclastic resorption is a first order process. Making in addition certain simplifying assumptions, which are shown to have a modest influence on the final results, a mean rate of bone resorption can be calculated using a development of the well known Stewart-Hamilton formula. The apposition rate is calculated as the sum of the resorption rate and the calcium balance. Augmentation and diminution, defined as equal and opposite exchange processes, are given by the difference between A and the apposition rate. 4. The results of our first thirteen studies in normal subjects and patients with metabolic bone disease are presented, together with analyses of some data from the literature. It is concluded that the development of an atraumatic method for measuring rates of bone formation and resorption in the whole body would be an important advance in the study of metabolic bone disease, and this work is presented so that critical comparisons may be initiated between this tracer method and independent histological methods for measuring these parameters.
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  • 77
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    Keywords: Electron spin resonance ; Crystallinity ; Bone ; Osteons
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    Notes: Summary Several groups containing 10–15 isolated osteons differing in their degree of maturity were analysed. Samples were isolated from undecalcified human and bovine bone sections. The crystallinity coefficient, defined as the ratio of the number of radiation-induced paramagnetic defects in the crystalline lattice of hydroxyapatite to the total ash content, was calculated. The results were compared with measurements performed on fragments of total cortical bone, primary periosteal bone, and inner circumferential lamellar bone. The results show a higher crystallinity of fully calcified osteons as compared with that found at the initial stage of calcification. No differences in the ash content were observed between human osteons, from different stages of calcification evaluated morphologically and by X-ray absorption. These differences were evident when bovine osteons differing in their stage of calcification were compared. Human fully calcified osteons contain 60% ash and their crystallinity coefficient is 52.1. Human osteons at the initial stage of calcification contain 57% ash and their crystallinity coefficient is 40.6. The same parameters for fully calcified bovine osteons and for bovine osteons at the initial stage of calcification are 59% ash, 62.6 crystallinity coefficient and 46% ash, 43.0 crystallinity coefficient, respectively.
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    Calcified tissue international 24 (1977), S. 65-72 
    ISSN: 1432-0827
    Keywords: Bone ; Strength ; Tension ; Microstructure ; Osteon
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Standardized human and beef femoral compact bone specimens were tested in tensile impact and the dynamic mechanical properties were determined. The microstructure of 45 beef and 47 human bone specimens were examined histologically to determine if there is a structural basis to account for strength differences in the bone samples. Strong negative correlations were obtained between the maximum stress and the percentage area of secondary osteons in each specimen. For human bone samples, the energy absorption capacity and the modulus of elasticity were also found to have strong negative correlations with the percentage area of secondary osteons present in each specimen. Linear regression equations were obtained describing the impact strength properties in terms of the percentage areas of secondary osteons and cavities in the samples. Fracture surfaces of the tested bone specimens were examined in a scanning electron microscope. Most surfaces exhibited a fairly rough texture indicating a quasi-cleavage type of failure. Fractographic analysis of bone fracture surface was helpful in understanding the micromechanics of bone fracture.
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  • 79
    ISSN: 1432-0827
    Keywords: Bone ; Density ; Composition ; Renal osteodystrophy ; Dogs
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    Notes: Summary Bone density and composition were studied in trabecular and cortical bone from control 2 and 4 year old beagles and those which had various degrees of renal failure as a result of perinatal irradiation. Changes in the two types of bone were qualitatively similar but consistently greater in trabecular bone. In a group identified as markedly uremic, the decrease (P〈0.025) from control levels in specific gravity was about 4 times greater in trabecular than in cortical samples. The decrease (P〈0.025) in grams of ash/ml was 9% in trabecular bone and 2% in cortical bone. These changes were associated with an increase (P〈0.001) in water content and, on a percent by volume basis, approximately equal decreases in ash (P〈0.025) and combustible matter (P〈0.025). In a mildly uremic group there were similar trends in mean values but the only significant difference was an increase (P〈0.001) in trabecular bone water.
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  • 80
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    Calcified tissue international 24 (1977), S. 201-208 
    ISSN: 1432-0827
    Keywords: Calcitonin ; Plasma calcium ; Radiocalcium ; Bone
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    Topics: Biology , Medicine , Physics
    Notes: Summary The effect of intravenous salmon calcitonin (SCT) infusion on the net rate of loss from plasma of recently injected45Ca has been studied in conscious fasted rats, some of which had been previously thyroparathyroidectomized (TPTX). The45Ca was injected 10 h prior to or 1 or 2 h after the initiation of the infusion. Salmon calcitonin (0.05–0.2 MRC mU/g body weight/h) infusion produced the expected hypocalcemia and hypophosphatemia. The drop in plasma calcium levels produced by the hormone was lowered further by the addition of phosphate to the infusate. Plasma45Ca changes, during SCT infusion, followed a biphasic pattern when the radionuclide injection preceded SCT infusion by 10 h. During the first part of the infusion, the rate of loss of45Ca from plasma was greater in SCT-treated than in control rats. Later, this pattern reversed, and the rate of loss of45Ca from plasma became slower in SCT-treated animals. If45Ca was injected after SCT infusion was initiated only the second phase of the biphasic effect was observed; i.e., disappearance of45Ca from plasma was slower in SCT-infused than in control rats. These data are considered to support the hypothesis that rapid and opposing calcium fluxes exist between bone fluid and plasma, and that calcitonin acts by decreasing the calcium efflux from bone fluid. The calcium moved in this efflux system includes some that has only recently entered bone fluid from plasma. This is the explanation given for the initial decrease in plasma45Ca concentrations following SCT infusion. The reverse effect in the second phase can be explained as the result of a secondary reduction in calcium influx to bone fluid from plasma.
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  • 81
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    Calcified tissue international 23 (1977), S. 113-114 
    ISSN: 1432-0827
    Keywords: Bone ; Absorption Spectra ; Collagen ; Hydroxyapatite
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Absorption spectra of human bone and its two major constituents (collagen and apatite) were obtained in the wavelength region extending from 2000 to 12,000 Å. In the last two cases a, minimum transmission (maximum absorption) was uniformly observed in the ultraviolet region. The two samples after exposure to ultraviolet radiations show a shift in the peak positions. Absorption peaks in the total bone spectra are not reproducible in its two constituents. After exposure to UV radiations the position of maximum absorption is displaced. A possible interpretation of the observed results is presented.
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  • 82
    ISSN: 1432-0827
    Keywords: Parathyroidectomy ; Diphosphonates ; Bone ; Collagen ; Hydroxyproline
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary Disodium ethane-1-hydroxy-1,1-diphosphonate (EHDP) may impair mineralization and reduce turnover of bone. However, its administration has been shown to lead to an accumulation of osteoid in both man and the rat. Using rats, the effects of surgical parathyroidectomy and administration of EHDP (30 mg EHDP/kg body weight by subcutaneous injection daily for 5 days) upon bone composition, uptake of3H-proline into hydroxyproline in bone and the subsequent urinary excretion of labelled and unlabelled hydroxyproline have been compared. Both parathyroidectomized (PTX) rats and EHDP-treated animals accumulated more hydroxyproline in bone and excreted less total urinary hydroxyproline than sham-PTX control animals, the changes induced by EHDP being of greater magnitude than those induced by surgical parathyroidectomy. Skeletal buildup of hydroxyproline was associated with decreased catabolism of mature collagen rather than enhanced synthesis of hydroxyproline in bone. EHDP administration, although decreasing urinary excretion of newly synthesised hydroxyproline, did not alter the skeletal synthesis of hydroxyproline as measured by uptake of3H-proline. In all parameters measured, responses to EHDP were similar in PTX and sham-PTX rats. It is concluded that depression of parathyroid hormone mediated bone turnover may have contributed to, but did not fully account for, the lowering of bone turnover and changes in bone composition induced by EHDP in sham-PTX rats in the present experiment.
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  • 83
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    Calcified tissue international 22 (1977), S. 27-33 
    ISSN: 1432-0827
    Keywords: Protein ; Bone ; Blood plasma ; Dentine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract The spectrum of plasma proteins present in human cortical bone and permanent dentine has been determined. One plasma glycoprotein, theαHS-glycoprotein, was found to be present at a high concentration in both bone and dentine and was shown to be concentrated in the mineralized tissues with respect to the other plasma proteins by factors of between 30 and 100. In this respect theαHS-glycoprotein is analogous to the G2B-glycoprotein and α-glycoprotein of bovine and rabbit b one, respectively. The binding ofαHS-glycoprotein and albumin to calcium phosphates generated within serum samples has been studied at different serum: precipitate ratios. In each case all theαHS-glycoprotein was removed from the samples and theαHS-glycoprotein was concentrated with respect to albumin by factors ranging from 370 at the highest serum: precipitate ratio to 25 at the lowest ratio. The plasmaαHS-glycoprotein concentrations of patients with Paget's disease of bone were shown to be substantially lower than the normal range, with significant negative correlation between theαHS-glycoprotein concentration and the plasma alkaline phosphatase activity.
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  • 84
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    Calcified tissue international 22 (1977), S. 183-189 
    ISSN: 1432-0827
    Keywords: Calcium ; Strontium ; Bone ; Mathematical modelling
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    Topics: Biology , Medicine , Physics
    Notes: Abstract 1. A method has been devised for comparing the impulse response functions of the skeleton for two or more boneseeking tracers, and for estimating the contribution made by measurement errors to the differences between any pair of impulse response functions. 2. Comparisons were made between the calculated impulse response functions for47Ca and85Sr obtained in simultaneous double tracer studies in sixteen subjects. Collectively the differences between the47Ca and85Sr functions could be accounted for entirely by measurement errors. 3. Because the calculation of an impulse response function requires fewera priori assumptions than other forms of mathematical analysis, and automatically corrects for differences induced by recycling of tracer and non-identical rates of excretory plasma clearance of tracer, it is concluded that differences shown in previous in vivo studies between the fluxes of*Ca and*Sr into bone can be fully accounted for by undetermined oversimplifications in the various mathematical models used to analyse the results of those studies.85Sr is therefore an adequate tracer for bone calcium in most in vivo studies.
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  • 85
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    Calcified tissue international 22 (1977), S. 219-226 
    ISSN: 1432-0827
    Keywords: Hydroxyapatite ; Bone ; X-ray Diffraction
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    Notes: Abstract Evidence relating to the existence of crystalline bone mineral in vivo is considered, and bone apatite crystal structure investigated using an x-ray powder diffraction technique. Specimens of femoral compacta excised post-mortem from male and female subjects ranging from 3 1/2 years to 87 years of age have been studied. Values of the ratio c/a of bone apatite crystal cell axes for females are significantly higher (p〈0.05) than for males. Moreover, significant change of c/a with age is observed for males (p=0.0005) but not for females (p=0.30). Differences in c/a are interpreted as indicating substitution of constituent ions in the bone apatite crystals.
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  • 86
    ISSN: 1432-0827
    Keywords: Diphosphonate ; Bone ; Osteoclasts ; Autoradiography
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Abstract Male rats weighing 100 g were injected with vehicle (control group), 0.4 or 4.0 mg/kg/day of ethane-1-hydroxy-1,1-diphosphonate (EHDP). Beginning after 6 days of EHDP treatment all rats received six subcutaneous injections of3H-thymidine at 8 h intervals. The multiple labeling index of autoradiographically defined labeled cells was determined for the osteoprogenitor cells of the primary spongiosa of the proximal tibial metaphysis. The osteoclasts in the proximal tibial metaphysis were assayed for population parameters and incorporation of labeled nuclei at sequential intervals after the administration of3H-thymidine. After 6 days of EHDP pre-treatment the multiple labeling index of the osteoprogenitor cells was reduced with 4.0 mg/kg/day dose of EHDP. At the beginning of the experimental kinetic sampling period, after the 6 days of EHDP pre-treatment, EHDP treatment resulted in significant increases in the numbers of osteoclasts, nuclei per osteoclast profile, and total osteoclast nuclei. During the sampling period, the 4.0 mg/kg/day dose of EHDP caused further increases in the number of total osteoclasts. EHDP treatment caused a marked increase in the rate of incorporation and accumulation of3H-thymidine-labeled nuclei in osteoclasts. Thus, EHDP causes an accelerated rate of osteoclast production from precursor cells in spite of its ability to slow bone resorption. Although the origin of osteoclast nuclei is not clear, the kinetic data from this study suggests that osteoclast nuclei are not arising from osteoprogenitor cells or osteocytes in the EHDP-treated animals.
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  • 87
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    Calcified tissue international 23 (1977), S. 103-112 
    ISSN: 1432-0827
    Keywords: Albumin ; Polyvinylpyrrolidone ; Tissue fluid ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The passage of tissue fluid through cortical bone has been investigated using radioactively labelled macromolecules as markers. The results suggest that in the cortex of young rabbit femur the movement of tissue fluid is in the same net direction as blood, mainly from the endosteal to the periosteal surface. Some albumin is incorporated from extravascular tissue fluid into calcified matrix at sites of bone formation. Polyvinylpyrrolidone, average molecular weight 35,000 is able to pass through extravascular tissue fluid in bone but isnot incorporated into calcified matrix. In rabbits made vitamin D deficient, much less albumin is retained in regions of bone formation than is the case with controls. Albumin adsorbs to the surface of calcium phosphate precipitates and it is suggested that this mechanism may be mainly responsible for its incorporation into bone.
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  • 88
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    Calcified tissue international 24 (1977), S. 143-149 
    ISSN: 1432-0827
    Keywords: Bone ; Periodontal ligament ; Bone resorption ; Aryl sulfatase ; Mineralization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The distribution of aryl sulfatase in the rat periodontal ligament was investigated by ultrastructural histochemistry. In the periodontal ligament aryl sulfatase was localized specifically in osteoclasts and in vicinal perivascular macrophages. Macrophages associated with bone formation did not stain. The authors interpret this specificity as a potential marker for osteoclast differentiation from macrophages — or as a further indication of the capacity of macrophages to modulate their enzymatic complement in response to the environment. To explain the absence of aryl sulfatase in areas of bone formation we suggest that different sulfate esters are mobilized from resorbing and mineralizing matrices, and that only the enzyme associated with bone resorption is histochemically detectable with the artificial substrates currently used.
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  • 89
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    Calcified tissue international 24 (1977), S. 243-248 
    ISSN: 1432-0827
    Keywords: Paracrystallinity ; Hydroxyapatite ; Bone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary X-ray diffraction patterns from oriented bone sections show that the crystalline apatite content of untreated mature cortical bovine bone has, in fact, a paracrystalline structure (i.e., no long range order). There is anisotropy in both lattice distortions and the sizes of the coherently diffracting domains. The paracrystalline mean distance fluctuations, (g) were found to be 1.5 (±0.1)% and 2.9 (±0.2)% for the basal and prism planes respectively, the corresponding paracrystalline sizes being 220 (±20) and 70 (±10) Å. The paracrystalline structure became more ordered above 600°C, suggesting the association of hydroxyl and possibly carbonate and other ions with the paracrystalline structure. The paracrystalline model for bone apatite helps explain anomalies between X-ray and electron microscope measurements of crystal size and also some of the biological functions of the crystalline apatite.
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  • 90
    ISSN: 1432-0878
    Keywords: Cholesterol ; Symbiotes ; Aphids ; Digitonin ; Autoradiography
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Pea aphid primary symbiotes have previously been shown to synthesize cholesterol in vitro. Two electron microscopic techniques were used here to determine whether the symbiotes also synthesize cholesterol in vivo and whether this cholesterol is made available to the aphid. We also inquired into a possible role of secondary symbiotes in cholesterol biosynthesis. Treatment of aphids with digitonin resulted in significant alteration of ultrastructural sites in primary and secondary symbiote membranes. We concluded that these sites are areas of high cholesterol concentration in the symbiotes. Electron microscopic autoradiography with 3H-mevalonate precursor indicated that both primary and secondary symbiotes synthesize cholesterol; in both cases, the majority of grains were associated with the symbiote membranes. While the frequency of grains on the symbiotes remained constant, irrespective of incubation time in labelled media, the frequency of grains over surrounding tissues increased exponentially as the time of incubation was increased from 30 min to 8 h, indicating that symbiote cholesterol is transported to other tissues. High voltage electron microscopic autoradiography permitted thick section autoradiography, reducing the time of emulsion exposure from 54 days (thin section) to 12 days (0.5 μm sections).
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  • 91
    ISSN: 1432-0878
    Keywords: CSF-contacting subependymal cells ; Frog hypothalamus ; Fluorescence microscopy ; Autoradiography ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Blue-green fluorescent subependymal cells with intraventricular processes were shown by the fluorescent histochemical method to be distributed from the preoptic recess to the infundibular recess of the frog hypothalamus. Electron microscopy revealed at least two types of CSF-contacting subependymal cells, type 1 containing large dense granules (about 100–200 nm in diameter) and type 2 containing small dense core vesicles (about 60–100 nm in diameter). Subsequent to fixation in permanganate solution, the small dense core vesicles in type 2 cells reacted with the fixative and consistently showed a dense content. However, the large granules in type 1 cells were mostly pale or less dense after this fixation. Two hours after intraventricular injection of 3H-dopamine, a large number of silver grains appeared only in the cytoplasm of intraventricular processes possessing dense core vesicles (type 2 cells). A few grains were also found in the perikarya. It is concluded that type 2 cells are catecholamine-storing cells. It is suggested that type 1 cells in the infundibular recess are peptidergic neurons which may secrete some hypothalamic regulating hormones of the anterior pituitary. Most of these cells in the preoptic recess belong to the neurosecretory cells of the preoptic nucleus, while some cells probably function similarly to those in the infundibular recess.
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  • 92
    ISSN: 1432-0878
    Keywords: Pituitary ; Dexamethasone ; ACTH ; Autoradiography ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary 3H-Dexamethasone (10 μg/kg) was injected intravenously in adrenalectomized rats and after survival times of 5, 30, 60, and 180 min its uptake within the pituitary was studied by autoradiography. Radioactivity was concentrated in cell nuclei in the pars nervosa and pars distalis. Within the pars intermedia, only cells of the marginal zone were labeled. In the pars distalis, some cells showed a weak nuclear accumulation of radioactivity as early as 5 min after injection. The tissue radioactivity was nearly maximal at 5 min, and the proportion of radioactivity in nuclei reached a maximum of 60–70% by 30 min. In competition experiments, non-radioactive steroids (1 mg/kg) were injected 5 min before 3H-dexamethasone and sacrifice was 30 min later. Dexamethasone markedly diminished the nuclear accumulation in the pars distalis, but corticosterone and progesterone did not. In the pars nervosa, corticosterone and progesterone competed for nuclear uptake of 3H-dexamethasone, although less effectively than dexamethasone itself. Different cell types in the pars distalis were characterized by treating autoradiograms with an immuno-peroxidase bridge procedure. Cells treated with anti-ACTH 17–39 had the greatest nuclear concentration of radioactivity, and those stained with anti-TSH were least heavily labeled. Cells treated with antisera to GH, PRL, and hCG were moderately labeled.
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  • 93
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    Cell & tissue research 177 (1977), S. 141-144 
    ISSN: 1432-0878
    Keywords: Cell cycle ; Intestinal mucosa (mouse) ; Autoradiography ; Transplantation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Transplantation of a small fragment of the ileum beneath the kidney capsule in syngenic mice results in the formation of a cyst lined with proliferating intestinal epithelium. The duration of the cell cycle in this epithelium was determined (using tritiated thymidine and the FLM method) as 14.5 h, as compared with 11.5 h in the intestinal epithelium in situ. We conclude that the intestinal content has little effect on the cell cycle of epithelial cells of the small intestine.
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  • 94
    ISSN: 1432-0878
    Keywords: Pineal gland, rabbit ; Organ culture ; Ultrastructure ; Indoleamine synthesis ; Autoradiography
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In order to localize pineal indoleamine synthesis at the ultrastructural level, an electron microscopic analysis was carried out on rabbit pineal tissue, cultured for several days in a medium containing the inhibitor p-chlorophenylalanine. Furthermore, electron microscopic autoradiography was applied to rabbit pineal tissue cultured in a medium containing the tritium labeled precursor 5-hydroxytryptophan. p-Chlorophenylalanine altered the ultrastructure of the mitochondria in the light pinealocytes only. Incubation with 3H-5-hydroxytryptophan caused a moderate and uniform labeling of the pinealocytes. These data suggest that the synthesis and storage of indoleamines in the rabbit pineal gland do not take place within the conventional membrane-limited cell organelles but more diffusely in the cytosol of the light pinealocytes. The spherical enlargement of the mitochondria in the light pinealocytes after culturing with p-chlorophenylalanine supports the biochemical findings of Hori et al. (1976) that the mitochondria are the carriers of the enzyme tryptophan 5-hydroxylase. The subsequent decarboxylation of 5-hydroxytryptophan to serotonin probably occurs free in the cytosol. The strong labeling of the noradrenergic nerve endings (in contrast to the cholinergic ones) after incubation with 3H-5-hydroxytryptophan demonstrates that the noradrenergic nerve endings in the rabbit pineal gland can, at least in vitro, selectively take up 5-hydroxytryptophan.
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  • 95
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    Medical & biological engineering & computing 15 (1977), S. 72-74 
    ISSN: 1741-0444
    Keywords: Bone ; Elasticity ; Mechanical strength
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Summary The mechanical properties of canine long bones have been determined. The mean and standard deviation of the maximum load supported by six fresh canine femurs in 3-point bending was 1366±253 N. In all cases, the bones sustained a considerable amount of plastic deformation before failure. The modulus of elasticity (GN/m2), ultimate tensile stress (MN/m2) and yield stress (MN/m2) were 13·86, 108·3 and 88·3 for six humeral specimens and 13·88, 142·1 and 104·0 for 13 tibial specimens, respectively. The tibial specimens showed a statistically significant higher ultimate strength than the humeral specimens.
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  • 96
    Electronic Resource
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    Medical & biological engineering & computing 15 (1977), S. 207-208 
    ISSN: 1741-0444
    Keywords: Bone ; Instrumentation ; Tensile strength
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
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  • 97
    Electronic Resource
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    Cell & tissue research 178 (1977), S. 333-339 
    ISSN: 1432-0878
    Keywords: Thymus, Human fetus ; Lymphocytes ; DNA synthesis ; Autoradiography ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Fragments and suspensions of human fetal thymus were incubated in the presence of 3H-TdR to permit study of the distribution and morphology of DNA-synthesizing cells. Results of light and EM autoradiography showed that 1. although DNA-synthesizing cells were present in the medulla, the vast majority of these cells were localized in the thymic cortex, 2. cells with the typical EM appearance of small lymphocytes and lymphoid blast cells both synthesized DNA, and 3. cells in S-phase were predominantly 8 to 12 μm in size.
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  • 98
    Electronic Resource
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    Cell & tissue research 183 (1977), S. 379-384 
    ISSN: 1432-0878
    Keywords: Brain ; Rat ; Diethylstilboestrol ; Autoradiography
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The cellular and subcellular localization of radioactivity in the brain of immature female rats was determined by dry-mount autoradiography 2 h after iv injection of 1.0 μg of (monethyl-3H) diethylstilboestrol per 100 g body weight. A specific topographic pattern of nuclear concentration of the synthetic oestrogen was obtained similar to that for 3H-oestradiol-17β in specific neurons of the basal hypothalamus, preoptic region and amygdala. In competition experiments, the nuclear concentration of radioactivity in all areas studied was inhibited by unlabeled oestradiol, while unlabeled testosterone had no effect. These data suggest that although oestradiol can bind to androgen receptors, the oestrogen receptor itself can account for the localization seen after the injection of 3H-oestradiol.
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  • 99
    Electronic Resource
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    Cell & tissue research 183 (1977), S. 221-237 
    ISSN: 1432-0878
    Keywords: Visual system (Rat) ; Autoradiography ; Quantitative analysis ; Suprachiasmatica nucleus ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Light microscopic autoradiography performed subsequent to intraocular injection of 3H-leucine revealed silver grains (SG) above axons of the optic tract which could be followed into the ventral and caudal portion of the suprachiasmatic nuclei (SCN) and above the contralateral anterior hypothalamic nucleus (AHN). By high resolution photometric measurement and computer processing the labelled areas were analysed, thus yielding statistical data of the relative grain distribution. The highest SG density was found in the ventrolateral part of both SCN (SCvl), confirming earlier reports concerning retinohypothalamic connections. That area exhibiting a cytoarchitecture different from the remaining nucleus was traversed, however, by numerous labelled axons. In the caudal part of both SCN a specific projection field of retinal fibres could be located. Here, almost no traversing fibres contribute to the rather circumscribed marked area. In the ventral part of the contralateral AHN, diffuse labelling well above background levels could be observed. Distinction between bypassing and terminating fibres within the SCvl could not be made using light microscopy. Analysis of SG distribution of the SCvl with electron microscopic autoradiography revealed a specific localization of SG within presynaptic terminals containing clear vesicles and pale mitochondria.
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  • 100
    ISSN: 1432-0878
    Keywords: Ovary (Rat) ; Luteinization ; LH(HCG) Receptors ; Autoradiography
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The LH(HCG) receptors in the ovaries of immature rats which were either untreated, or primed with PMSG and HCG, have been studied with a histochemical method which has proved to be as effective as when earlier used in the rat testis. This method, which consists of the topical application of 125I-HCG to picric acid-formaldehyde (PAF) fixed frozen sections followed by autoradiography, is also suitable for quantitative studies on the distribution of receptors. In the ovary of the immature 26 days old rat, the LH(HCG) receptors are localized exclusively in the interstitial and thecal tissues. After PMSG treatment many receptors appear in the granulosa of the large antral follicles. These receptors are most numerous in the outer layers of cells and least numerous in the inner. At the same time there are fewer receptors in the thecal and interstitial cells which have undergone the process of luteinization. After PMSG and HCG treatment the newly formed corpora lutea have few receptors, but these become progressively more numerous on subsequent days. It is suggested that, in the rat, the luteinization of the ovarian LH-target cells is associated with an initial decrease in the number of their LH(HCG) receptors.
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