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PubMed2001

Effects of chelating agents and sodium hypochlorite on mineral content of root dentin.

Effects of combined and single use of EDTA, RC-Prep, and NaOCl on mineral content of root dentin were evaluated in vitro using energy dispersion spectrometric microanalysis. Thirty-six standardized midroot dentin specimens obtained from human anterior teeth were used. Specimens were polished and divided into six experimental groups. The first two groups were treated with EDTA or RC-Prep followed by NaOCl irrigation. Groups 3 to 5 were treated with EDTA, RC-Prep, and NaOCl, respectively. The last group was irrigated with saline solution as a control. Levels of calcium, phosphorus, and magnesium were measured in the root dentin after treatments. The results showed that (i) EDTA combined with NaOCl irrigation as final flush and NaOCl alone changed the calcium/phosphorus ratio of root dentin significantly (p < 0.05); and (ii) there was a significant increase in the magnesium level after the use of chelating agent combined with NaOCl (p < 0.05). It was concluded that using NaOCl irrigation as final flush altered the effectiveness of chelating agents on root dentin.

Calcium↗
PubMed1985

Electron microprobe analysis of human labial gland secretory granules in cystic fibrosis.

X-ray microanalysis of freeze-dried labial gland cryosections revealed that Na concentration was doubled and the Ca/S concentration ratio was decreased in secretory granules of labial glands from patients with cystic fibrosis (CF) when compared with glands from normal subjects. Other results suggested that the decrease in the Ca/S concentration ratio resulted from an increase in S concentration. These findings imply that mucous granules in labial saliva showed a CF-related increase in Na and S content, and such changes would be expected to affect the rheology of the mucus after exocytosis. In contrast with a previous study in human parotid glands, no evidence was found for CF-related changes in cytoplasmic or nuclear Na, K, and Ca concentrations. Significant elemental differences were found between secretory granules and nuclei and cytoplasm of control cells.

Adolescent↗
PubMed1979

Accumulation of cadmium and lead in the gills of Mytilus edulis: X-ray microanalysis and chemical analysis.

The accumulation of Cd and Pb in the gills of the lamellibranch mollusc Mytilus edulis has been studied by electron microscopy, X-ray microanalysis, atomic absorption spectroscopy and radionuclide monitoring. The patterns of accumulation of the two elements differ markedly as do the sites of deposition whithin the gills. Lead is found extracellularly as crystalline deposits in the basal lamina which forms the capillary walls of the gill lamellae. The Pb is found associated with Ca in equiatomic ratios and occurs either as a mixed or complex carbonate. Cadmium is always associated with S and frequently with P in membrane bound vesicles within the cells of the gill epithelium and in the amoebocytes. The S is probably attributable to the presence of cysteine residues in a metal binding protein which can be extracted from the gills. Analysis of the metal binding protein shows that it binds Ag, Cd, Cu, Fe, Hg, Sn and Zn. Its amino acid composition is similar to that reported for eels and limpets but has a lower cysteine content than mammalian metal binding protein.

Amino Acids↗
PubMed1988

Cis DDP in combination with selenium and sulfur. Subcellular effect in kidney cells. Electron microprobe study.

Cis DDP is an anticancer agent used in the treatment of diverse cancers, but its use has been limited by its nephrotoxic effect. We sought to modify this major toxic side effect by the addition of S2O3-- (sodium thiosulfate) or SeO2 (selenium dioxide) or both. In the present study, only ultrastructural and X-ray microanalysis data are reported. No kidney lesions were observed after the administration of SeO2 and Cis DDP in contrast to Cis DDP with S2O3-- or with S2O3-- plus selenium. An organelle of the kidney cell, the lysosome, has a particular role in the concentration of mineral elements in this cell. Platinum was observed in the lysosome along with sulfur, after the administration of Cis DDP and S2O3. Platinum, selenium and sulfur were observed after administration of SeO2, S2O3 and Cis DDP. No mineral deposits were observed after administration of SeO2 and Cis DDP. The role of sulfur seems to be very different from that of selenium. We hypothesize that sulfur favors the intralysosomal concentration of platinum or that selenium associates with platinum. The fact that selenium is not reabsorbed by the kidney cell seems to favor the hypothesis of urinary elimination of platinum. The present work confirms our previous study concerning the role of SeO2 in combination with Cis DDP.

Animals↗
PubMed2002

Immobilization of human thrombomodulin to expanded polytetrafluoroethylene.

BACKGROUND: The success of synthetic grafts for vascular reconstruction remains limited by thrombosis and intimal hyperplasia. In addition to the well-described antithrombotic effects of thrombomodulin, we have demonstrated that recombinant human thrombomodulin (rTM) inhibits arterial smooth muscle cell proliferation induced by thrombin. This study investigated the binding of functional rTM to expanded polytetrafluoroethylene (ePTFE). METHODS: Immobilization of rTM was achieved by either (1) a direct coating or (2) a two-step binding process using a water-soluble condensing cross-reaction agent EDAC to modify the ePTFE surface followed by binding of rTM. The samples were then subjected to a tangential shaken wash. The evidence of bound rTM was evaluated by both morphologic and functional studies. RESULTS: SEM, BSI, and X-ray microanalysis identified that the two-step binding method resulted in significantly greater binding of rTM molecules to ePTFE pre- and post a 7-h wash than the direct coating method. With the two-step binding method rTM ranging from 0.25 to 12.5 microg immobilized to ePTFE-activated protein C (APC) in a concentration-dependent manner by more than 6000-fold compared to the buffer control (P < 0.04) and 50-85% more than direct coating (P < 0.004). With direct coating, the level of APC dropped significantly to near 40% of the preshaken level at 2 h and diminished to 26% at 7 h. Whereas, the level of APC with the two-step binding stabilized at 51 and 49% after being shaken 2 and 7 h, respectively. CONCLUSION: Functional rTM binding to ePTFE was significantly improved with a new two-step binding method.

Coated Materials, Biocompatible↗
PubMed1988

X-ray microanalysis of cartilage and chondrocytes.

The elemental content of differentiating chondrocytes in situ and in vitro was studied by X-ray microanalysis. The in situ studies were carried out on semi-thick cryosections and on thin sections of freeze-dried low-temperature vacuum-embedded rat rib growth plate. Resting, proliferative, and hypertrophic cells were analyzed separately. The in vitro studies were carried out on cultured cell fractions isolated from rat rib growth plate. The cell fractions were relatively homogeneous and contained resting - early proliferative, proliferative or hypertrophic cells, respectively. Data from cells in vitro are generally not comparable to data obtained from cells in situ. Chondrocytes in situ contain relatively high levels of Na, which is probably due to: (1) high Na levels in the cartilage matrix formed intracellularly, and (2) low supply of oxygen and nutrients. High Na levels in (some of) the proliferative cells may also be related to mitotic activity. The chondrocyte matrix contains relatively high levels of K. The data in the present study are compared to those of previous studies on the elemental content of chondrocytes.

Animals↗
PubMed1990

Quantitative X-ray microanalysis of alveolar macrophages after long-term treatment with amiodarone.

Treatment with the iodine-containing antiarrhythmic drug, amiodarone, can cause pulmonary toxicity. Alveolar macrophages are particularly susceptible to formation of lipidrich lamellar bodies in amiodarone-treated animals. Amiodarone and several of its metabolites accumulate in the cell. Previously, we have reported that the technique of X-ray microanalysis is useful in monitoring the distribution of iodine in freeze-dried cryosections of alveolar macrophages from Fischer 344 rats 24 hr after a single dose of amiodarone. In the present study, we examine the effects of longer term amiodarone treatment of 1 or 9 weeks. Substantial changes in iodine distribution occur in the cells with increasing length of drug treatment. High concentrations of iodine are found early in the lamellar bodies. The iodine levels in the nuclei slowly increase with the length of treatment, and after 9 weeks of treatment, approach those found in the lamellar bodies. It is possible that this accumulation of iodine in the nuclei is due to the presence of polar metabolites. In addition, the potassium concentration in the cell decreases and the sodium increases with treatment duration. These changes in cations are most likely due to altered ion transport in the macrophages by the inhibition of membrane Na-K-ATPase by the drug and its principal metabolite, desethylamiodarone.

Amiodarone↗
PubMed2000

Localization and fate of aluminium in the digestive gland of the freshwater snail Lymnaea stagnalis.

The digestive gland of the freshwater snail Lymnaea stagnalis, exposed to water containing an elevated concentration of aluminium at neutral pH for up to 30 days, followed by a 20 day recovery period, was examined by light and electron microscopy and X-ray microanalysis. Aluminium was localized in the yellow granules present in the digestive and excretory cells and in the green and small granules present in the digestive cells. More aluminium, silicon, phosphorus and sulphur were present in all three granule types from aluminium exposed snails. The number of yellow and green granules from the digestive gland of aluminium exposed snails showed a progressive increase over the experimental period compared to controls. The number and aluminium content of the granules is likely to reflect the role of the digestive gland as a 'sink' for accumulated aluminium. We propose that intracellular monomeric silica is involved in the detoxification of aqueous aluminium which at neutral pH is largely in the form of an insoluble polyhydroxide. The increased amounts of sulphur and phosphorus in the granules are likely to be part of a broad response to metal loading but probably do not play a significant role in the storage and detoxification of aluminium.

Aluminum↗
PubMed1989

Expression of anionic sites on tumour cells at different stages of tissue invasion in vivo: a comparative study by X-ray microanalysis.

Quantification of colloidal iron hydroxide (CIH) labelling by X-ray microanalysis was used to investigate anionic sites at the surface of Ehrlich carcinoma cells from different locations in the mouse host. Individual tumour cells from peritoneal ascites suspensions (pre-invasion stage) varied up to threefold in their ability to bind CIH and a similar degree of intra-tumour heterogeneity was observed in different experimental animals. Pretreatment of the cells with neuraminidase confirmed that binding was at least partly due to surface sialic acid. Invasive cells isolated from mesenteric tumour nodules were also heterogeneous with regard to the availability of surface anionic sites, as were tumour cells adhering to the surface of the mesentery; however, in both these populations CIH binding was significantly greater on average than for free ascites tumour cells. The results suggest that surface anionic sites are determinants of the invasiveness of malignant cells in vivo, and that both the number and topography of these sites may be important in modulating tumour cell behaviour.

Animals↗
PubMed1995

Energy-dispersive X-ray microanalysis of calcified tissues after NaOCl treatment and/or resin embedding.

By energy-dispersive X-ray microanalysis (EDX) in scanning electron microscopy, we studied the calcification of the inner-basic lamellas of a cow bone and the coronal cementum of a horse tooth treated with sodium hypochlorite (NaOCl). These tissues were divided into 4 groups with a combination of NaOCl treatment and polyester-resin embedding including, [A]: non-NaOCl + Resin, [B]: non-NaOCl + non-resin, [C]: NaOCl + Resin, and [D]: NaOCl + non-Resin. From the Ca and P values by EDX analysis, it was suggested that the natural porous spaces of [B] were higher than those of [A], and both the natural and NaOCl-soluble porous spaces were highest in [D]. However, [A] had the lowest porous spaces in both the tissues because the micropores formed 3-dimensionally by NaOCl treatment might be incompletely filled with the resin. The backscattered electron microscopy and the difference of the Ca/P ratios indicated that the NaOCl treatment of the calcified tissues caused some minerals besides organic materials to dissolve. Thus, the sample preparation of [B] is suitable for the quantitative EDX of calcified tissues, whereas the data of [C] except for the Ca/P ratio may be used to approximate the Ca and P contents.

Animals↗
PubMed1995

Characterization of thin hydroxyapatite layers formed on anodic titanium oxide films containing Ca and P by hydrothermal treatment.

An anodic titanium oxide film containing Ca and P (AOFCP) was formed on commercially pure titanium which was anodized in an electrolytic solution of dissolved beta-glycerophosphate (beta-GP) and calcium acetate (CA). Hydroxyapatite (HA) crystals were precipitated by hydrothermally heating the AOFCP at 300 degrees C. After hydrothermal treatment, the film was characterized by scanning electron microscopy (SEM), energy-dispersive X-ray microanalysis (EDX), and tensile tests. The morphology, composition, and amount of HA crystals precipitated were significantly affected by the composition of the electrolytes. Near-stoichiometric HA crystals with high crystallinity were precipitated completely covering the AOFCP surface at specific electrolyte concentrations. The HA layers were thin at 1-2 microns in thickness. The adhesive strength of the film increased with decreasing electrolyte concentration and the maximum value was about 40 MPa. In vitro tests for 300 days suggested that the stability of the film was high. The high adhesive strength may result from the AOFCP existing as an intermediate layer between the HA layer and a titanium substrate. The intervention of the AOFCP may have prevented abrupt changes in Ca and P content at an HA coating-titanium interface as seen in a plasma-sprayed one. The porous TiO2 matrix of the AOFCP may be suitable for nucleation sites of HA crystals, as well as SiO2 matrix of silicate bioactive glasses or glass ceramics.

Acetates↗
PubMed1987

Scanning transmission ion microscopy as it complements particle induced x-ray emission microanalysis.

The early uses of Scanning Transmission Ion Microscopy (STIM) with MeV ions are reviewed. The transformation of STIM energy-loss images into maps of areal density is discussed, and is illustrated with images of a fruit fly head (Drosophila melanogaster). Freeze-dried male heads are transparent to 4-MeV protons in the dorsal and frontal directions, but in the sagittal direction the brain is opaque. STIM with molecular ions is shown to be useful for increasing contrast in low density areas. For recording registered STIM and PIXE images without changing accelerator parameters, apertures are used to accomplish the required change in beam intensity (a factor of 10(5]. Molecular ions are used to assess contamination of the microbeam by scattered ions. Pixel by pixel ratios of x-ray intensity to areal density are taken to obtain maps of element concentrations. Calcium and iron maps are shown. Inner parts of the fly head are clearly seen in the concentration maps. The PIXE exposure caused differential displacements of inner parts of the head by 15 micron or less. Weight loss during the PIXE exposure was measured to be 3%.

Animals↗
PubMed1984

Age-dependent decrease of the passive Rb+ and K+ permeability of the nerve cell membranes in rat brain cortex as revealed by in vivo measurement of the Rb+ discrimination ratio.

Young, adult and old male CFY rats (2, 12 and 24 mth of age, respectively) were treated with a daily dose of 30 mg RbCl/100 g body weight, in form of aqueous solution injected intraperitoneally for 14 days. A considerable part of the intracellular K+-content of the body was replaced by Rb+ during this treatment. After cessation of the RbCl injections, a relative steady state came into being in each age group, called Rb+-release period. During this period Rb+ and K+ contents of the blood serum and the cisternal CSF were measured by atomic absorption spectrophotometry, and of the intracellular space of brain cortical cells by energy-dispersive X-ray microanalysis. Ultrastructural features of the brain cortex were also checked by transmission electron microscopy. For X-ray microanalysis, the L-line of Rb at 1.694 keV energy was used at 10 kV accelerating voltage in a scanning electron microscope equipped with an EDAX System F. Rb+ and K+ concentrations were obtained for the cellular dry mass and converted into wet concentrations on the basis of intracellular water contents known from former experiments. Rb+-replacement of K+ did not cause any ultrastructural alteration in the brain cortex. However, the Rb+ accumulation displayed a very significant age-dependent increase: at the beginning of release, adult and old rats had 32.6 and 44.7 mM Rb+ in their intracellular water as against the 8.6 mM found in the young group, and similar proportional difference persisted during 20 days of the release. Rb+ discrimination ratios (DR) calculated either for the blood or the CSF displayed very considerable age-dependent increase: the values of the adult and old groups were 191 and 242% of the young one, indicating that the passive Rb+ (and K+) permeability of the nerve cell membrane decreases throughout the life span of rats. These results give further support to the membrane hypothesis of aging.

Aging↗
PubMed1997

Changes in surface levels of mercury, silver, tin, and copper of dental amalgam treated with carbamide peroxide and hydrogen peroxide in vitro.

OBJECTIVES: The effect of 10% carbamide peroxide or 10% hydrogen peroxide on the surface levels of mercury, silver, tin, and copper of amalgam fillings was tested in vitro with scanning electron microscopy and energy dispersive spectrometric microanalysis. STUDY DESIGN: Samples of amalgam were treated for 14 and 28 days with either 10% carbamide peroxide or 10% hydrogen peroxide solutions and compared with phosphate buffer controls. RESULTS: A significant increase in mercury levels occurred after treatment with carbamide peroxide for 14 days (p < 0.01) and 28 days (p < 0.001) and after treatment with hydrogen peroxide for 28 days (p < 0.001). A significant increase in silver levels occurred after treatment with carbamide peroxide for 14 days (p < 0.05) and 28 days (p < 0.01) and subsequent to treatment with hydrogen peroxide for 14 days (p < 0.05) and 28 days (p < 0.001). A significant reduction in tin levels occurred after treatment with hydrogen peroxide for 14 days (p < 0.01) and 28 days (p < 0.001), and after treatment with carbamide peroxide for 28 days (p < 0.01). A significant reduction in copper levels was found after treatment with carbamide peroxide for 14 days (p < 0.05). CONCLUSIONS: It appears that prolonged treatment with bleaching agents may cause microstructural changes in amalgam surfaces, possibly increasing exposure of patients to toxic byproducts.

Carbamide Peroxide↗
PubMed1986

Elemental concentration gradients between subcellular compartments.

To gain information on the mechanisms involved in the establishment and maintenance of subcellular gradients of Na, K, Cl and other elements in the flagellate, Euglena gracilis, we turned to the technique of ultracentrifugal stratification of its intracellular contents, which is achieved without loss of viability or cell rupture. Stratified and non-stratified Euglena were cryofixed for energy-dispersive X-ray microanalysis of Na, K, Cl and other elements in thin freeze-dried cryosections. A number of significant elemental concentration differences (expressed as mmol kg-1 dry weight) were found between chloroplast, nucleus, paramylon granules and open cytoplasm (which contained ribosomes, membranes and macromolecules associated with the cytomatrix) in the non-stratified cells. Stratification caused several ions to be redistributed. For example, we observed a significant increase in K and Cl in the nucleus, which was correlated with the condensation of chromatin. Also Cl, but not Na, decreased significantly in the region of cytoplasm that was cleared of observable ribosomes, membranes and macromolecules associated with the cytomatrix, as well as of observable cytochemical enzyme activity. We conclude from the data that more than half of the Cl in open cytoplasm was adsorbed to or entrapped in material that was removed by ultracentrifugation. Thus, it appears that a close association of at least one ion, Cl, with ultracentrifugable material is involved in maintenance of the measured Cl concentration in the open cytoplasm of the non-stratified cell.

Animals↗
PubMed2000

X-ray microanalysis of cultured keratinocytes: methodological aspects and effects of the irritant sodium lauryl sulphate on elemental composition.

Irritant substances have been shown to induce elemental changes in human and animal epidermal cells in situ. However, skin biopsies are a complicated experimental system and artefacts can be introduced by the anaesthesia necessary to take the biopsy. We therefore attempted to set up an experimental system for X-ray microanalysis (XRMA) consisting of cultured human keratinocytes. A number of methodological aspects were studied: different cell types, washing methods and different culture periods for the keratinocytes. It was also investigated whether the keratinocytes responded to exposure to sodium lauryl sulphate (SLS) with changes in their elemental composition. The concentrations of biologically important elements such as Na, Mg, P and K were different in HaCaT cells (a spontaneously immortalized non-tumorigenic cell line derived from adult human keratinocytes) compared to natural human epidermal keratinocytes. The washing procedure and time of culture influenced the intracellular elemental content, and rinsing with distilled water was preferred for further experiments. Changes in the elemental content in the HaCaT cells compatible with a pattern of cell injury followed by repair by cell proliferation were seen after treatment with 3.33 microM and 33 microM SLS. We conclude that XRMA is a useful tool for the study of functional changes in cultured keratinocytes, even though the preparation methods have to be strictly controlled. The method can conceivably be used for predicting effects of different chemicals on human skin.

Cells, Cultured↗
PubMed2004

Redistribution of thorotrast into a liver allograft several years following transplantation: a case report.

Thorotrast was used as a radiographic contrast agent in the United States from about 1930 to the mid-1950 s. Its use was discontinued when it was recognized that its radioactivity caused long-term deleterious effects. Such long-term sequelae of intravascular Thorotrast injection include, most notably, hepatic and hematologic malignancies and hepatic fibrosis. Some patients who had received Thorotrast subsequently received liver transplants. However, it was not known whether or not Thorotrast could become redistributed within the new allograft. A single report in 1994 demonstrated that radioactivity was detected by gamma-ray spectroscopy in liver allografts shortly after transplantation. No report has identified Thorotrast in the allografts of long-term transplant survivors, and the redistribution of Thorotrast into allografts has not been documented histologically or by electron microscopy. We report a case of recurrent Thorotrast into a liver allograft 10 years post-transplant. We evaluated the native liver and allograft specimens for the presence of thorium utilizing light microscopy, electron microscopy and electron X-ray microanalysis. This case report demonstrates for the first time the redistribution of Thorotrast into a long-surviving liver allograft using histologic, electron microscopic and X-ray microanalysis techniques.

Cholangitis, Sclerosing↗