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PubMed1990

The chemoreceptor surface of the taste disc in the frog, Rana esculenta. An ultrastructural study with lanthanum nitrate.

The distribution of lanthanum on the taste disc of the frog, Rana esculenta, after en bloc staining of the tongue with lanthanum nitrate was studied at the ultrastructural level by means of scanning electron microscopy (in the secondary electron mode or in the back scattered electron mode), energy-dispersive X-ray microanalysis, transmission electron microscopy and electron spectroscopic imaging. It was consistently found that lanthanum distribution on the surface of the taste disc is not homogeneous and that the surface of putative receptor cells is in contact with strongly lanthanum-positive material. Calcium co-localizes with lanthanum at that level. These results suggest that different microenvironments exist at the surface of the taste disc and that this could be relevant to the receptor function.

Animals↗
PubMed2003

Effects of bioactive glass particles and their ionic products on intracellular concentrations.

Numerous studies have described the bioactive properties of glass particles in the SiO(2)-CaO-Na(2)O-P(2)O(5) system. This kind of material is capable of developing a direct contact with bone through dissolution and physicochemical reactions. We have investigated the influence of bioactive particles, and ionic products from the same particles, on the intracellular concentrations in monocyte cells, which are among the first cells to colonize implantation sites. The only way to access these concentrations and particularly diffusible ionic concentrations (potassium, sodium, and chlorine) is to use cryomethods coupled to electron probe microanalysis. We have paid particular attention to the potassium:sodium ratio, the most sensitive criterion of viability. We have cultured cells with bioactive glass particles and in a conditioned medium obtained from the dissolution of the glass particles in the standard medium. Our study demonstrates that cells cultured in a conditioned medium are more active than cells cultured in a standard medium, or cells exposed to bioactive particles, and particles are more toxic for cells than are ionic products.

Biocompatible Materials↗
PubMed1980

Freeze-dried, plastic-embedded tissue preparation: a review.

The freeze-dried, plastic-embedded specimen is a versatile type of animal soft tissue preparation, with attributes that commend it for some types of analytical and morphological studies. The preparation involves rapid freezing, freeze-drying, osmic acid vapor fixation, and embedding in an epoxy resin. This preparation, though difficult and tedious, offers some advantages for quantification and localization of water-soluble constituents in intracellular spaces. When prepared according to established protocol, there is evidence to suggest that the embedded sample faithfully retains intracellular levels of water-soluble constituents that are present at time of cryofixation. The sample can be stored easily and indefinitely and the tissue can be examined in a variety of ways. Thin sections will accept many stains for light and electron microscopy; the sample can be used for some histochemical procedures; and quantification of intracellular electrolytes is possible with electron probe microanalysis using one of several techniques. A major disadvantage of the method is the lack of satisfactory preservation of normal solute distributions in extracellular spaces. Important considerations for instruments equipped with Si(Li) energy detector systems are the decrease in mass fraction of the elements of interest due to embedding material, and complications in analysis for some elements because of the presence of osmium. This latter problem is of little consequence for analysis by wavelength spectrometers; and the increase in sample density, because of the embedding material, can be used to advantage for thick sample analysis.

Freeze Drying↗
PubMed1998

Bioactive bone cement: effect of the amount of glass-ceramic powder on bone-bonding strength.

We examined the influence of the proportion of glass-ceramic powder in a bioactive bone cement of our formula on the bone-bonding ability of cement. Changes in cement bonding with time also were examined. The bioactive bone cement consisted of MgO-CaO-SiO2-P2O5-CaF2 glass-ceramic powder (AW-GC powder) and bisphenol-alpha-glycidyl methacrylate (Bis-GMA)-based resin. AW-GC powder was added to the cement as 0%, 30%, 50%, 70%, and 80% w/w. Rectangular plates (2 x 10 x 15 mm) of each cement with polished surfaces were implanted into the proximal metaphysis of the tibiae of male rabbits, and the failure load was measured by detaching tests 10 and 25 weeks after implantation. The failure loads of each cement were 0% = 0.03, 30% = 1.52, 50% = 2.67, 70% = 3.56, and 80% = 5.59 kg at 10 weeks, and 0% = 0.05, 30% = 1.68, 50% = 2.77, 70% = 3.80, and 80% = 6.37 kg at 25 weeks. Observation of the cement-bone interface revealed that all bioactive bone cements (30%-80%) formed direct contact with bone whereas intervening fibrous tissue was observed in all specimens of the 0% group. By scanning electron microscopy, all bioactive bone cements (30%-80% groups) showed direct contact with bone at the cement-bone interface. In the 0% group, direct contact with bone at the cement-bone interface was not observed. By electron-probe microanalysis, a Ca-P-rich layer was not detected at the cement-bone interfaces of the 30%-70% bioactive bone cements, but in some samples of the 80% cement specimens a thin Ca-P-rich layer (3 microns thick) was observed at the interface at 10 and 25 weeks after implantation. These results show that all of the bioactive bone cements tested had the ability to bond to bone and to function as bioactive composites of ceramics and polymers.

Animals↗
PubMed2007

A new approach for the recovery of precious metals from solution and from leachates derived from electronic scrap.

A new approach is described for the recovery of precious metals (PMs: Au, Pd and Ag) with >99% efficiency from aqueous solution utilising biogas produced during the aerobic growth of Klebsiella pneumoniae. Gold was recovered from electronic scrap leachate ( approximately 95%) by this method, with some selectivity against Cu. The recovered PM solids all contained metal and sulphur as determined by energy dispersive X-ray microanalysis (EDX). X-ray powder diffraction analysis (XRD) showed no crystalline metal sulphur compounds but a crystalline palladium amine was recorded. Silver was recovered as a sulphide (found by EDX), carbonate and oxide (found by XRD). EDX analysis of the Au-precipitate showed mainly gold and sulphur, with some metallic Au(0) detected by XRD. The gold compound was shock-sensitive; upon grinding it detonated to leave a sooty black deposit.

Aerobiosis↗
PubMed1983

Copper storage in the liver of the wild mute swan (Cygnus olor). Its possible relation to pollution of harbor waters by antifouling paints.

Postmortem examination of three wild mute swans (Cygnus olor) from a harbor area disclosed an unusual black discoloration of the liver. Chemical, histochemical, and microscopic studies, along with electron-probe microanalysis, showed that cytoplasmic pigment granules in the liver cells contained a copper-protein complex. Similar findings have been reported in Danish and English studies on large numbers of wild mute swans. Two control mute swans from The Bronx Zoo had negligible amounts of hepatic copper. The striking difference between the wild and the captive swans in hepatic copper content suggests that the copper in the wild swans was of environmental origin, most likely from copper-rich antifouling paint used extensively in the marine industry. Flakes of this paint may be ingested by swans searching for food in the sediment of harbor waters.

Animal Population Groups↗
PubMed1988

Characterization of endosteal osteoblastic cells isolated from mouse caudal vertebrae.

We have developed a reliable procedure for isolating endosteal osteoblasts from mouse trabecular bone. Endosteal osteoblasts were obtained by migration and proliferation of the cells from the metaphyseal bone surface of caudal vertebrae onto nylon meshes. The isolated cells were cultured in Dulbecco's modified Eagle's medium supplemented with 10% fetal calf serum. The cell population consisted of 95% alkaline-phosphatase-positive cells. The cell level of alkaline phosphatase was elevated (1.19 +/- 0.26 (SD) mumol PNP/mn/mg protein) and the enzyme activity was heat-inhibitable, indicating its skeletal origin. Light and electron microscopic observation revealed that cells have morphologic and ultrastructural appearance of typical osteoblasts with high protein synthesis activity. Osteoblasts grown in multilayers in the presence of 50 micrograms/ml ascorbic acid produced within 4 days an abundant fibrous intercellular collagenous matrix forming nodules in which osteocyte-like cells were embedded. Immunolabeling revealed synthesis of type I collagen but no detectable type III collagen. In presence of 7 mM beta-glycerophosphate the matrix became mineralized after 14-21 days of culture. Mineralization could not be induced by mouse skin fibroblasts cultured under similar conditions. The mineral deposits were closely associated with the collagen matrix, consisted of EDTA-removable, Von Kossa and alizarin red S stainable material and were composed of hydroxyapatite crystals identified by X-ray electron probe microanalysis. The isolated endosteal osteoblasts also displayed an intense (+457%) increase in intracellular cAMP production in response to human (1-34) PTH (2 x 10(-8) M) stimulation. The confluent cells responded to 20 nM 1,25(OH)2D3 by a significant 45% reduction in heat labile alkaline phosphatase activity. This procedure allowed us to isolate from trabecular bone a cell population that differentiates into osteoblasts in vitro, respond to calcitropic hormones and that retains its capacity to form a calcified bone tissue in culture. This method provided us a culture system for investigating the differentiation and metabolism of endosteal osteoblastic bone forming cells.

Animals↗
PubMedDate not supplied

Silicone breast implants: pathology.

Questions as to the bioreactivity of silicone breast implants (SBIs) have recently been intensely scrutinized, most notably by the media and legal system. Pathologists must be aware of the controversy and treat each SBI and associated tissue as a potential lawsuit. Grossly, silicone is a clear, viscous substance that may be observed either within or extruding from a silastic bag. By light microscopy, silicone is a nonstainable, nonpolarizable, refractile substance. Thicker sections, especially when viewed by non-Köhler illumination, phase-contrast, and darkfield microscopy will enhance visualization. Ultrastructurally, silicone is an electron-dense, amorphous substance often located within phagocytic vacuoles or extracellularly within the stroma. Correlating electron probe microanalysis allows for reliable identification. In most cases, a fibrous capsule surrounds the SBI, with the interface lining varying from a virtually acellular to a synovial-like lining composed of phagocytic and secretory cells. Silicone can often be identified within the fibrous capsule and also in distant tissues biopsied for suspected autoimmune disorders, such as synovium, skin, and lymph nodes, often without ultrastructural evidence of cytologic effects. This study has demonstrated that silicone accumulates at distant tissue sites due to preexisting inflammation acting as a stimulus. Thus, silicone is not a primary inducer of inflammatory disease processes. These findings are supported by various large epidemiologic studies.

Animals↗
PubMed1995

Subcutaneous nodules in a patient hyposensitized with aluminium-containing allergen extracts: a microanalytical study.

Electron spectroscopic imaging (ESI) is a recent tool for electron microscopic analysis which permits the recognition of the distribution of elements in a specimen, with a spatial resolution up to 5 nm. The authors performed the ESI together with X-ray microanalysis (EDX) and transmission electron microscopy to study granulomatous subcutaneous nodules in a patient who had undergone a desensitising therapy with an aluminium-containing vaccine. Aluminium was detected both by EDX and ESI, mainly in lysosomes of histiocytes containing needle-shaped material in a lucent matrix. However, ESI provided a better localization of aluminium with respect to EDX. This result suggests that the identification of aluminium-containing structures cannot be obtained only by ultrastructural morphology and underlines the utility of a microanalitycal study for a correct diagnosis in the presence of a needle-shaped deposition of dense material in lysosomes.

Aluminum↗
PubMed2003

Charge-related problems associated with X-ray microanalysis in the variable pressure scanning electron microscope at low pressures.

In variable pressure scanning electron microscopy (VPSEM) the current data suggests that considerable caution is required in the interpretation of X-ray data from nonconductive samples, depending on the operating conditions. This article reviews some of the documented approaches and presents data that illustrate the nature and magnitude of the effects of charge above, on, and in the sample on the detected X-ray emissions from the sample and from elsewhere within the VPSEM specimen chamber. The collection of reliable and reproducible X-ray data has been found to require relatively high specimen chamber gas pressures, at the upper end of or beyond the available pressures for most VPSEMs. It is also shown that sample characteristics, including composition, strongly influence local charge effects, which can significantly affect the primary electron landing energy and consequently the resultant emitted X-ray signal under low pressure environments.

Electron Probe Microanalysis↗
PubMed1981

Staining of oral epithelium with the zinc iodide-osmium reaction.

Some of the parameters affecting the staining of keratinized oral epithelium with the zinc iodide-osmium reaction were examined using light and electron microscopy and electron probe microanalysis. Factors examined were block size, incubation temperature and the effect of aldehyde prefixation. Large blocks (4 mm cube) were subdivided after incubation and the staining of the centre and edge compared. Generally the reaction was more variable at the edge than in the centre. Small block (1 mm cube) showed a more intense reaction when incubated at 24 degrees C than at 4 degrees C. In all these preparations, final reaction product was seen over Golgi systems, lysosome-like bodies, membrane-coating granules and, in the more intensely stained regions, over endoplasmic reticulum and nuclear membranes as well. In prefixed material, mitochondria were frequently stained in addition to the other organelles. Energy dispersive analysis showed the reaction product to be similar in all preparations and to contain high levels of zinc and osmium but not iodine.

Animals↗
PubMedDate not supplied

Measures for spectral quality in low-voltage X-ray microanalysis.

Characteristic x-ray production with energetic electrons depends strongly on the overvoltage, the ratio of the incident beam energy to the critical excitation energy for the atomic species of interest. Low-voltage x-ray microanalysis (beam energy < or = 5 keV) is especially susceptible to artifacts due to sample charging because the overvoltage is low and even slight charging can strongly affect peak intensities. The Duane-Hunt bremsstrahlung limit is a good diagnostic to detect sample charging. Dynamic charging effects, however, can influence spectra despite an apparently satisfactory Duane-Hunt limit. Dynamic charging effects must be examined by time series experiments, or through use of dynamic energy windows continuously measuring count rates placed across the spectrum. When charging is a problem, conductive surface coatings can eliminate the effects. When pristine surfaces must be examined without coating, the use of a conductive grid can control charging so that useful x-ray spectra can be obtained.

Artifacts↗
PubMed1998

Changes in the extracellular matrix on the surface of sintered bovine bone implanted in the femur of a rabbit: an immunohistochemical study.

The interface of implanted True Bone Ceramics (TBC; sintered bovine bone; Koken, Tokyo, Japan) was examined. In the primary experiment, TBC was implanted into the bone marrow of a rabbit's femur. The extracellular matrices (types I, II, and III collagens and fibronectin) of decalcified specimens collected 1-48 weeks postoperatively were immunohistochemically examined. Undecalcified sections collected 6 weeks postoperatively were used for line analyses of calcium and phosphorus, by a scanning electron microscope-electron probe microanalysis (SEM-EPMA) method. In a secondary experiment, TBC was implanted into an osteochondral defect of a femoral condyle, harvested 1-12 weeks postoperatively, and decalcified to examine the extracellular matrices at the interface. In the bone marrow in the early phase, TBC had absorbed quantities of fibronectin. Immature bone (containing both types I and III collagens) in direct apposition to the ceramic surface had matured (containing type I collagen alone) in the TBC pores. SEM-EPMA revealed the continuity of high levels of calcium and phosphorus at the TBC-bone interface. In the secondary experiment, enchondral ossification or fibrous tissue formation was observed near the articular surface. However, in the subchondral layer, direct bone formation was observed in the TBC pores. It was concluded that TBC has excellent bioactivity for inducing maturation of new bone matrix on porous surfaces.

Animals↗
PubMed2000

Long-term changes of hydroxyapatite-coated dental implants.

There are many controversies about the long-term prognosis of hydroxyapatite (HA)-coated implants. Failure may be related to compositional and structural changes of the coating occurring during implantation. Two retrieved and two unused HA-coated blade-type implants were examined by stereomicroscopy, secondary electron microscopy, Fourier transform infrared spectroscopy, X-ray diffraction, and electron probe microanalysis. The objective was to investigate the HA morphology, composition, and structure, and to characterize the changes that occurred in the retrieved implant coatings. Retrieved implants presented partial loss of the coating, especially at the apical and mesiodistal edges. Remaining HA was thick and flattened in the cervical and central areas and gradually thinner and rougher towards the apical and mesiodistal edges. Increase of Cl and Mg, decrease of OH, and X-ray diffraction peak broadening were found in the retrieved implant coatings, in comparison with the unused implants. Morphological changes of the retrieved implants seem to depend on stress values in the surrounding bone and on implant mobility. Compositional changes and increased amount of lattice imperfections appeared in the retrieved implant coatings, as a result of ion substitutions in the apatite lattice. However, the present study could not confirm the influence of these changes on implant failure.

Coated Materials, Biocompatible↗
PubMed1984

In vitro biodegradation of chrysotile fibres by alveolar macrophages and mesothelial cells in culture: comparison with a pH effect.

The modification of the chemistry of asbestos chrysotile fibres (Mg3(Si2O5)(OH)4) after their ingestion by cultured cells has been studied. Two types of cells involved in asbestos related pulmonary disease were used, rabbit alveolar macrophages (AM), recovered by bronchoalveolar lavage, and pleural mesothelial cells (PMC) obtained from the rat parietal pleura. Chemical characterisation of intracellular fibres was performed on unstained ultrathin sections by electron probe microanalysis. The results showed a progressive leaching of Mg, characterised by a time dependent decrease of Mg/Si. AM were more efficient than PMC at leaching intracellular chrysotile fibres since it took longer to obtain the same proportion of leached fibres with PMC than with AM. As in vitro Mg-leaching can be obtained by acid treatment, chrysotile fibres were incubated, either untreated or pretreated with cell membranes, at pH 4 or 7 for various times. The data show that the kinetic of leaching by AM was comparable with leaching at pH 4. The leaching by PMC was of the same order as leaching at pH 7. When membranes were adsorbed on to the fibres, a delayed leaching was observed. The results indicate that the solubilisation of chrysotile by AM could be an intraphagolysosomal event due to a pH effect. With PMC, however, it is not possible to draw this conclusion since nothing is known about the intracellular pH.

Animals↗
PubMed2003

Electron-probe X-ray microanalysis of individual particles of solid snow sediment with size factor correction.

A technique is proposed for X-ray electron-probe microanalysis (EPMA) of individual particles of solid snow sediment. Two sample preparation procedures were used to analyze the sediments. The PAP-method and the original bi-exponential model were used for matrix correction and the calculation of elemental composition. A method of calculating composition has been developed for approximately spherical individual particles, comparable in size with an area of X-ray generation. An analytical expression is proposed for determining the composition of these particles, taking account of their size. Including the size factor reduces the error of composition determination from 0.5-45 to 0.2-22% relative percent, for particles in the size range 1-3 microm.

Journal Article↗
PubMedDate not supplied

Black and brown pigment gallstones differ in microstructure and microcomposition.

The two subtypes of pigment gallstones, black and brown stones, differ in chemical composition and pathogenesis. We examined a black bilirubinate stone and a black phosphate stone (which represented opposite ends of the compositional spectrum of black noncarbonate stones), a black carbonate stone, and a brown pigment stone using scanning electron microscopy and microchemical techniques to determine if stone microstructure and microcomposition reflected different patterns of formation. The cross-sectional surfaces of the black bilirubinate and black phosphate stones were smooth and homogenous. Electron probe microanalysis demonstrated high concentrations of sulfur and copper in the center of the black bilirubinate stone; sulfur was in a low valence state consistent with disulfide linkages in proteins. The brown stone was rough-surfaced with lamellated bands on cross-section. The lighter-colored bands in this stone contained virtually all of the detected calcium palmitate, while the darker sections contained much more calcium bilirubinate. Plasma oxygen etching demonstrated a network of protein interdigitating with calcium bilirubinate salts in the black bilirubinate and black phosphate stones but not in the black carbonate or brown stones. Argon ion etching demonstrated that calcium bilirubinate was in a closely packed rod-shaped arrangement in all three black stones but not in the brown stone. We conclude that the marked differences in structure and composition between the black noncarbonate and brown pigment gallstones support the hypothesis that the two major pigment gallstone types form by different mechanisms. In addition, the layered structures of the black carbonate and brown stones suggest that stone growth is affected by cyclic changes in biliary composition.

Argon↗
PubMed2003

Reduction of potassium tellurite to elemental tellurium and its effect on the plasma membrane redox components of the facultative phototroph Rhodobacter capsulatus.

Anaerobically light-grown cells of Rhodobacter capsulatus B100 are highly resistant to the toxic oxyanion tellurite (TeO(3)(2-); minimal inhibitory concentration, 250 microg/ml). This study examines, for the first time, some structural and biochemical features of cells and plasma membrane fragments of this facultative phototroph grown in the presence of 50 microg of K(2)TeO(3) per ml. Through the use of transmission microscopy and X-ray microanalysis we show that several "needlelike" shaped granules of elemental tellurium are accumulated into the cytosol near the intracytoplasmic membrane system. Flash-spectroscopy, oxygen consumption measurements, and difference spectra analysis indicated that membrane vesicles (chromatophores) isolated from tellurite-grown cells are able to catalyze both photosynthetic and respiratory electron transport activities, although they are characterized by a low c-type cytochrome content (mostly soluble cytochrome c(2)). This feature is paralleled by a low cytochrome c oxidase activity and with an NADH-dependent respiration which is catalyzed by a pathway leading to a quinol oxidase (Qox) inhibited by high (millimolar) concentrations of cyanide (CN(-)). Conversely, membranes from R. capsulatus B100 cells grown in the absence of tellurite are characterized by a branched respiratory chain in which the cytochrome c oxidase pathway (blocked by CN(-) in the micromolar range) accounts for 35-40% of the total NADH-dependent oxygen consumption, while the remaining activity is catalyzed by the quinol oxidase pathway. These data have been interpreted to show that tellurite resistance of R. capsulatus B100 is characterized by the presence of a modified plasma-membrane-associated electron transport system.

Anaerobiosis↗