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PubMed1981

Distribution of calcium in the suctorian Discophrya collini: an x-ray microanalytical study.

Discophrya collini is a free-living suctorian with tentacles which can be induced to contract by means of a range of experimental stimuli, including the application of CaCl2 and MgCl2 but not BaCl2. X-ray microanalysis of glutaraldehyde-only fixed cells shows Ca to be present in the cytoplasmic ground substance and elongate dense bodies (EDB). In 10(-1) M CaCl2-treated cells, calcium levels remain unchanged except for a three-fold increase in the EDB. Treatment of cells with 10(-1) M MgCl2 and 10(-1) M BaCl2 does not result in their detection in the cell. It is suggested that EDB may act as reservoirs controlling levels of calcium.

Animals↗
PubMed1992

Comparative surface microanalysis of failed Brånemark implants.

The chemical composition and topography of an implant surface determine the human immunologic system response. This study compared the surfaces of 13 Brånemark oral implants, 11 that came from retrieved specimens which failed initially or did not osseointegrate and 2 that were never implanted (controls). The period of implantation in human jaws varied between 3 and 20 months. After cleaning and sterilization, the topography, surface chemical composition, and thickness of the oxide layer were studied. The results obtained with scanning electron microscopy did not show any significant topographic differences among the specimens. X-ray spectrographic microanalysis showed very similar composition (titanium and amounts less than 0.5% of other elements) in the outermost layer of the analyzed specimens. The Auger spectroscope revealed considerable percentage differences in the amount of carbon and silicon in the last monolayers, which could be attributed to handling or to an inadequate cleaning process. This places the retrieved specimens out of the acceptable statistical limits of contamination by introducing a factor of doubt for long-term prognosis in the hypothetical situation of their re-use.

Dental Implants↗
PubMed1999

Antimicrobial activities of synthetic bismuth compounds against Clostridium difficile.

Clostridium difficile is a major nosocomial pathogen responsible for pseudomembranous colitis and many cases of antibiotic-associated diarrhea. Because of potential relapse of disease with current antimicrobial therapy protocols, there is a need for additional and/or alternative antimicrobial agents for the treatment of disease caused by C. difficile. We have synthesized a systematic series of 14 structurally simple bismuth compounds and assessed their biological activities against C. difficile and four other gastrointestinal species, including Helicobacter pylori. Here, we report on the activities of six compounds that exhibit antibacterial activities against C. difficile, and some of the compounds have MICs of less than 1 microgram/ml. Also tested, for comparison, were the activities of bismuth subcitrate and ranitidine bismuth citrate obtained from commercial sources. C. difficile and H. pylori were more sensitive both to the synthetic bismuth compounds and to the commercial products than were Escherichia coli, Pseudomonas aeruginosa, and Proteus mirabilis, and the last three species were markedly resistant to the commercial bismuth salts. Testing with human foreskin fibroblast cells revealed that some of the synthetic compounds were more cytotoxic than others. Killing curves for C. difficile treated with the more active compounds revealed rapid death, and electron microscopy showed that the bismuth of these compounds was rapidly incorporated by C. difficile. Energy dispersive spectroscopy X-ray microanalysis of C. difficile cells containing electron-dense material confirmed the presence of internalized bismuth. Internalized bismuth was not observed in C. difficile treated with synthetic bismuth compounds that lacked antimicrobial activity, which suggests that the uptake of the metal is required for killing activity. The nature of the carrier would seem to determine whether bismuth is transported into susceptible bacteria like C. difficile.

Anti-Bacterial Agents↗
PubMed1999

Effects of static or dynamic mechanical stresses on osteoblast phenotype expression in three-dimensional contractile collagen gels.

Studies performed at tissular (three-dimensional, 3-D) or cellular (two-dimensional, 2-D) levels showed that the loading pattern plays a crucial role in the osteoblastic physiology. In this study, we attempted to investigate the response of a 3-D osteoblastic culture submitted to either no external stress or static or dynamic stresses. Rat osteosarcoma cells (ROS 17/2.8) were embedded within collagen type I lattices and studied for 3 weeks. Entrapment and proliferation of cells within the hydrated collagen gel resulted in the generation of contractile forces, which led to contraction of the collagen gel. We used this ability to evaluate the influence of three modes of mechanical stresses on the cell proliferation and differentiation: (1) the freely retracted gels (FRG) were floating in the medium, (2) the tense gels (TG) were stretched statically and isometrically, with contraction prevented in the longitudinal axis, and (3) the dynamic gels (DG) were floating gels submitted to periodic stresses (50 or 25 rpm frequency). Gels showed maximum contraction at day 12 in 50 rpm DG, followed by 25 rpm DG, then FRG (88%, 81%, 70%, respectively) and at day 16 in TG (33%). The proliferation rate was greater in TG than in FRG (+52%) but remained low in both DGs. Gel dimensions were related to the collagen concentration and on a minor extent to cell number. Cells in DG appeared rounder and larger than in other conditions. In TG, cells were elongated and oriented primarily along the tension axis. Scanning electron microscopy (SEM) showed that tension exerted by cells in TG led to reorientation of collagen fibers which, in turn, determined the spatial orientation and morphology of the cells. Transmission electron microscopy (TEM) performed at maximum proliferation showed a vast majority of cells with a distended well-developed RER filled with granular material and numerous mitochondria. Alkaline phosphatase activity peaked close to the proliferation peak in FRG, whereas in TG, a biphasic curve was observed with a small peak at day 4 and the main peak at day 16. In DG, this activity was lower than in the two other conditions. A similar time course was observed for alkaline phosphatase gene expression as assessed by Northern blots. Regardless of the conditions, osteocalcin level showed a triphasic pattern: a first increase at day 2, followed by a decrease from day 4 to 14, and a second increase above initial values at day 18. Microanalysis-x indicated that mineralization occurred after 14 days and TEM showed crystals within the matrix. We showed that static and dynamic mechanical stresses, in concert with 3-D collagen matrices, played a significant role on the phenotypic modulation of osteoblast-like cells. This experimental model provided a tool to investigate the significance and the mechanisms of mechanical activity of the 3-D cultured osteoblast-like cells.

Alkaline Phosphatase↗
PubMed1989

[Pathogenesis of disorders of transparency of the optic media of the eye. III. Quantitative detection of copper in relation to the ultrastructure of the vitreous body in chalcosis].

The authors elaborated a new analytical micromethod for the detection of elements in morphologically investigated ultrastructures of the vitreous body. For the investigation of quantitative ultrastructural changes electron microanalysis (EMA) was used, combined with raster electron microscopy (REM). Using this method, the authors proved ultrastructural changes of the vitreous body by selective uptake of copper in the trabecular collagen fibres of the vitreous body in experimental chalcosis. From the results ensues that copper does not diffuse in the vitreous body in a homogeneous manner but aggregates in irregular agglomerations of precipitates and crystals. The different quantitative detection of copper in different parts of the same specimen is according to the authors due to the heterogeneous affinity of ultrastructural portions of the vitreous body for copper.

Animals↗
PubMed2006

Acid-etching effects in hypomineralized amelogenesis imperfecta. A microscopic and microanalytical study.

OBJECTIVES: The purpose of this study was to use quantitative x-ray microprobe analysis with scanning electron microscopy to define the morphostructural and calcification patterns in the enamel of teeth with the hypomineralized variant of amelogenesis imperfecta. STUDY DESIGN: We compared 5 fragments of permanent human canines from patients with clinically diagnosed hypomineralized amelogenesis imperfecta and 5 normal permanent canines from subjects without amelogenesis imperfecta. All specimens were etched with phosphoric acid for morphological and microanalytical examination. RESULTS: Two types of etching patterns were found; in addition, islets of pattern I were seen within areas of pattern II. Microanalysis detected no significant differences in calcium concentration between specimens with amelogenesis imperfecta and normal control specimens after acid etching. Pattern III was not observed. CONCLUSIONS: The changes and their distribution in the enamel structure after 30 s of acid etching are described in teeth with this rare disorder. Although these data seem to coincide with alterations in prism development, no alterations in calcium concentration were found.

Acid Etching, Dental↗
PubMed1995

Morphological effects of glycosaminoglycans on calcium oxalate monohydrate crystals.

The effects of individual glycosaminoglycan (GAG) species on calcium oxalate monohydrate (COM) crystal growth were studied in vitro by the observation of crystal morphology grown in a supersaturated calcium oxalate solution in the presence of GAGs using optical and scanning electron microscopes. GAGs affected the morphology of COM crystals differently depending on the species. The growth rates of the crystals formed in the presence of chondroitin-6-sulfate (ChS-C) were higher in length and lower in width and thickness than those of control crystals. The incorporation of dermatan sulfate or heparin into the crystals formed in the presence of these GAGs was revealed by X-ray microanalysis, whereas ChS-C was not detected in the crystals grown with it. The experiment using dicarboxylates, as a simple model of GAG molecules, showed that a distance between the side groups was important for their morphological effects. These findings suggested that the different effects of GAGs on the crystal morphology resulted from the differences in their interaction modes with COM crystal faces, that is, the differences in their binding behavior, their inhibition modes of crystal growth, and other roles played after binding to the crystals.

Calcium Oxalate↗
PubMed1984

Shell formation and calcium transport in the barnacle Chthamalus fragilis.

The mantle epithelium of the barnacle Chthamalus fragilis (Darwin) exhibits several ultrastructural features which may serve to regulate the calcification process. At the basis-mural plate and intermural plate junctions where rapid shell growth occurs, cells are characterized by long apical cytoplasmic projections and large intercellular spaces. These features may increase the functional surface area of the epithelium and enable more rapid deposition of calcium. The cells underlying the general shell surfaces contain numerous electron-dense inclusion bodies and show frequent cellular disintegration near the growing shell interface. Release of the granular contents of these inclusion bodies has been observed in both disintegrating and non-disintegrating cells. X-ray microanalysis revealed significantly higher calcium levels in the inclusion bodies than in the surrounding cytoplasm. This suggests a calcium transport role for these inclusion bodies. Cellular debris produced as a result of the disintegration of the mantle cells near the shell may play some role in the formation of the organic matrix of the shell. The presence of large numbers of mitochondria and well-developed apical microvilli in the cells of the inner mantle epithelium suggest that these cells serve to transport calcium into the mantle from the ambient sea water.

Animals↗
PubMed1990

Transmission electron microscopic characterization of hypersensitive human radicular dentin.

Transmission electron microscopy (TEM) and x-ray microanalysis (XMA) were used for the study of the ultrastructure of the lumens of dentinal tubules in superficial layers of dentin specimens obtained by use of a new biopsy technique from both hypersensitive and naturally desensitized areas of exposed root surfaces, in vivo. The TEM images showed clearly that the lumens of most of the tubules were occluded with mineral crystals in naturally desensitized areas, but such lumens were empty and surrounded with peritubular and intertubular dentin in hypersensitive areas. Moreover, electron-dense structures that lined peritubular dentin were observed in the empty lumens of dentinal tubules.

Adult↗
PubMed2004

Cementum-like tissue deposition on the resorbed enamel surface of human deciduous teeth prior to shedding.

Prior to the shedding of human deciduous teeth, odontoclastic resorption takes place at the pulpal surface of the coronal dentin, and this resorption occasionally extends coronally from the dentinoenamel junction into the enamel. After the end of resorption, however, the resorbed enamel surface is repaired by the deposition of a cementum-like tissue. Using this phenomenon as an observation model, in this study we examined the sequence of cellular and extracellular/matrix events involved in the enamel resorption repair by light and electron microscopy. As the odontoclast terminated its resorption activity, it detached from the resorbed enamel surface; thereafter, numerous mononuclear cells were observed along the resorbed enamel surface. Most of these mononuclear cells made close contact with the resorbed enamel surface, and coated pits or patches were observed on their plasma membrane facing this surface. Furthermore, they frequently contained thin needle- or plate-like enamel crystals in their cytoplasmic vacuoles as well as secondary lysozomes. Following the disappearance of these monononuclear cells, the resorbed enamel surface now displayed a thin coat of organic matrix. Ultrastructurally, this organic layer was composed of a reticular and/or granular organic matrix, but contained no collagen fibrils. Energy-dispersive X-ray microanalysis of this thin organic layer in undecalcified sections revealed small spectral peaks of Ca and P. Cementum-like tissue initially formed along this thin organic layer, increased in width, and appeared to undergo mineralization as time progressed. The results of our observations demonstrate that regardless of type of matrix of dental hard tissues, tooth repair may be coupled to tooth resorption, and suggest that mononuclear cells and an organic thin layer found on the previously resorbed enamel surface may play an important role in the repair process initiated after resorption of the enamel.

Dental Cementum↗
PubMed1996

Morphometry, scanning electron microscopy and X-ray spectral microanalysis of protostylid pits on human lower third molars.

Protostylid pits on 50 lower third molars were studied. The depth and width of the pits, the thickness of the enamel at the pit bottom and the concavity of the dentino-enamel junction under the pit were measured by light microscopy. The pit content was examined by scanning electron microscopy and X-ray spectral microanalysis. The results show that the morphometric variables of protostylid pits are similar to those of occlusal fissures and pits. The protostylid pits were seen to contain globular formations, belonging mainly to dental calculus; organic rod-shaped bodies, which could represent remnants of the enamel organ--or more probably--foreign bodies, were frequently observed. The study suggests that the development of the most common form of protostylid is basically similar to that of an occlusal cusp, with variations attributable to the low expression of the protostylid and particularities of cusp development in the lateral tooth wall.

Calcium↗
PubMed1994

Silicate pneumoconiosis in pigs: optical and scanning electron microscopical investigations with X-ray microanalysis.

Four cases of silicate pneumoconiosis are described in pigs raised near several chalk quarries and two cement works. The pulmonary changes were characterized by thickened alveolar septa, resulting in distorted airspaces, and small foci of initial fibrosis. In the bronchiolar and alveolar sites, as in the interstitium, free and intracytoplasmic dust was detected. An energy dispersive X-ray microanalysis coupled with a scanning electron microscope revealed that this dust was composed mainly of silicon, calcium, potassium, sulphur, aluminium and iron. In lung-associated lymph nodes, severe lymphoid cell depletion and dilatation of peritrabecular and subcapsular sinuses were constant findings. The inorganic material found in the lymph nodes contained the elements listed above. Air samples from the same geographical area revealed particulate pollutants, the qualitative features of which were similar to those found in lung and lymph nodal tissue. It is concluded that domestic animals raised in polluted environmental conditions represent an important biological source from which helpful data may be obtained for assessing risks to human health.

Animals↗
PubMed2004

Distribution of minerals in quinoa (Chenopodium quinoa Willd.) seeds.

The distribution of minerals in quinoa (Chenopodium quinoa Willd.) seed was examined using energy dispersive X-ray microanalysis (EDX) in combination with scanning electron microscopy (SEM). Phosphorus, K, and Mg coincided in localization in embryonic tissue. Since phytin globoids have been known to localize in protein bodies in embryonic cells of quinoa seed, it is thought that P is attributed to phytic acid and that K and Mg form to phytate. Calcium and K were present in the pericarp, where the cell wall is thickly developed, suggesting that these minerals are associated with pectin. Sulfur occurred in embryonic tissues, which would be derived from sulfur amino acid residues of storage proteins concentrated in the tissues. Abrasion of quinoa seeds resulted particularly in decrease in Ca content.

Calcium↗
PubMed1980

Genetic dissection of the final exocytosis steps in Paramecium tetraurelia cells: cytochemical determination of Ca2+-ATPase activity over performed exocytosis sites.

In different Paramecium tetraurelia strains the occurrence of a Ca2+-ATPase (or p-nitro-phenylphosphatase) activity at the preformed attachment and exocytosis sites of the secretory vesicles (trichocysts) was analysed by electron-microscopic cytochemistry and X-ray microanalysis. In conjunction with freeze-fracture studies it was found that only those strains, which contain rosette particles, display this Ca2+-ATPase activity (7S, K 40I, nd 9 (18 degrees C), while other strains (nd 6, nd 9 (27 degrees C), tam 38) are devoid of both these characteristics. The presence (absence) of rosette particles and of Ca2+-ATPase activity at the preformed exocytosis sites is correlated with the capability (incapability) of performing exocytosis in these strains. We discuss several possible interpretations of this structure-function correlation.

Animals↗
PubMed1982

Calcium-sequestering vesicles and contractile flagellar roots.

The intracellular localization of calcium during contraction of striated flagellar roots of the green alga Tetraselmis subcordiformis was investigated using calcium pyroantimonate and calcium oxalate cytochemistry and energy-dispersive X-ray microanalysis. Vesicles (0.25 - 0.50 microns diameter) located in the anterior-most region of the cell are calcium-sequestering organelles. These vesicles exhibit a consistent and reproducible localization of electron-opaque pyroantimonate and oxalate salts of calcium. Striated contractile flagellar roots (CFR) show a periodic pattern of calcium antimonate (Ca-Sb) deposits following 'calcium shock' stimulation of contraction of this organelle. When CFRs are in a fully extended state they do not show Ca-Sb deposits. Deposits of Ca-Sb precipitate are present throughout the cytosol when CFRs are stimulated to contract. The implications of these findings with respect to the regulation of intracellular levels of 'free' Ca2+ and to the mechanism of CFR contraction are discussed.

Calcium↗
PubMed1994

Measurement of BBN-induced alterations in rat urothelium by electron microscopic X-ray microanalysis.

The aim of this study was to analyse N-butyl-n-butanol-4-nitrosamine (BBN)-induced alterations of the urothelium in rats concerning its content of phosphorus, sulphur, chlorine, potassium and calcium using electron microscopic X-ray microanalysis (REM analysis). The following histopathological findings of the bladder mucosa were discovered after exposure to BBN: normal urothelium (n = 36); focal epithelial proliferations (n = 12) following 6-12 week's exposure; epithelial hyperplasia (n = 8) after urothelial carcinoma (n = 4) following 12 weeks' exposure. The observed phosphorus/sulphur and phosphorus/calcium ratios based on REM analysis did not show any statistical correlation with the morphological changes classified by light microscopy. Our data do not support the hypothesis raised by other investigators that an increase in phosphorus content or phosphorus/sulphur or phosphorus/calcium ratio could indicate early neoplastic transformations of urothelial cells as "tumor markers".

Animals↗
PubMed1993

The ultrastructure of copper amalgam-covered dentin from human deciduous teeth.

The structure of dentin subjacent to Cu-amalgam restorations was studied. The restorations had been inserted 7-8 years before extraction of the teeth. No lining had been applied. The dentin subjacent to the restorations had a greenish-grayish discoloration, and in the pulpal end irregular secondary dentin had formed. Energy-dispersive X-ray microanalysis of areas showing discoloration confirmed the presence of Cu. Semithin and ultrathin undemineralized sections were studied. Vital pulp tissue and irregular secondary dentin were observed. Close to the predentin and halfway into the dentin most of the dentinal tubules were occluded. In the control material only an occasional occluded tubule was seen. Close to the Cu-amalgam restoration some tubules were occluded or partly occluded, and some were open. The odontoblasts had responded to the irritating agent by obturation of the tubules, thus reducing the permeability of the dentin. Furthermore, irregular secondary dentin had formed in another attempt to wall off the irritating agent.

Child↗
PubMed2005

Destructive and non-destructive microanalysis of biocarbonates applied to anomalous otoliths of archaeological and modern sciaenids (Teleostei) from Peru and Chile.

Anomalous otoliths were discovered among modern and archaeological (8th millennium BP) sciaenids. The two species concerned, Cilus gilberti and Sciaena deliciosa, are common on the Peruvian-Chilean coast and do not seem to be affected by this morphological anomaly that maintained in their populations for thousands of years. The carbonates of the anomalous forms, determined by X-ray diffraction, are different from that of the normal otoliths, i.e. calcite and vaterite instead of aragonite. A method of non-destructive analysis by cathodoluminescence is tested and assumptions on the origin of the anomaly and its possible implications on environmental studies are advanced.

Animals↗