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PubMed1996

Ultrastructural analysis of the integument during the moult cycle in Ligia italica (Crustacea, Isopoda).

The formation of the cuticle was investigated during moulting of the isopod crustacean Ligia italica. The intermoult cuticle is a four-layered lamellar structure composed of chitin-protein fibrils and mineralized in its upper half. The distribution of calcium carbonate in cuticle during moult cycle was determined by cytochemical methods and X-ray microanalysis, Epi-and exocuticle are secreted during premoult. Calcium is resorbed from the old cuticle and accumulates in the ecdysal gap as calcium granules. Endocuticle is secreted after moult when the mineralization of exocuticle starts. The shape and ultrastructure of epithelial cells change during cuticle secretion and mineralization. Mitochondria, bundles of filaments, calcium granules and large amounts of glycogen accumulate in the apical cytoplasm of cells in premoult animals.

Animals↗
PubMed1989

Carbohydrate-controlled precipitation of apatite with coprecipitation of organic molecules in human saliva: stabilizing role of polyols.

Addition of common dietary carbohydrates to Millipore-treated human whole saliva either enhances or inhibits the formation of salivary precipitates, some carbohydrates showing no effect. The purpose of this study was to investigate the precipitation conditions more thoroughly and to elucidate the chemical nature of the precipitates formed. D-Xylose either enhanced precipitation (in long-term incubations) or had no appreciable effect (in 10 minute incubations). Other aldo- and keto-sugars and disaccharides (maltose, sucrose, lactose) generally enhanced precipitation, whereas all polyols (xylitol, D-sorbitol, mannitol, and maltitol) retarded the formation of turbidity in saliva. Xylitol inhibited formation of precipitates also in the presence of D-xylose, dextrans, and starch. Fast protein liquid chromatography (FPLC) of EDTA-soluble pellets obtained by centrifugation of the precipitates produced two major protein fractions (I and II) with a molecular weight of 112,000 and 46,000, respectively. The carbohydrates exerted a selective effect on the relative size of I and II in that polyol incubations resulted in a I to II ratio of 1:3, whereas control incubations (without added sugars) and incubations with other carbohydrates gave ratios of 1:6 to 1:10. Both peaks contained large amounts of acidic amino acids, proline, and glycine. The saliva precipitates contained a substantial portion of a crystalline phase that had the crystal structure of apatite, the individual crystallites being extremely small (less than 1 micron) with a Ca:P ratio of 1.46. The carbohydrates had a similar effect on the overall inorganic composition of the precipitates, but they had a clearly selective effect on the rate of formation of precipitates and on the relative amount of coprecipitating salivary proteins. This selectivity indicates that these carbohydrates, when consumed habitually, may exert different effects on the precipitation of Ca-salts at mineral-deficient enamel and dentine sites.

Adult↗
PubMed2006

Lithogenesis: induction of renal calcifications by nanobacteria.

Nanobacteria have been isolated from kidney stones and it has been suggested that they may act as a nucleus for the initiation of the renal stones. In the present study, we examine their role in biocrystallization and their in vivo effects on kidney pathology. Calcium oxalate monohydrate (COM) assay was carried out in the presence of nanobacteria to study biocrystallization. Wistar rats were given an intravenous injection of nanobacteria and the kidneys were examined for pathological changes. The COM assay showed accelerated biocrystallization of (14)C-oxalate in the presence of nanobacteria, indicating them to be efficient candidates for biomineralization. Histopathological studies revealed bacteria induced renal tubular calcifications and various manifestations of infection. Our studies confirm that nanobacteria may be involved in the pathogenesis of renal tubular calcification.

Animals↗
PubMed1991

Ceftriaxone-associated gallbladder sludge. Identification of calcium-ceftriaxone salt as a major component of gallbladder precipitate.

Ceftriaxone, a third-generation cephalosporin, is partially excreted into bile. With its clinical use, the formation of gallbladder sludge detected by ultrasonography has been reported. Four surgical specimens were examined and no gallstones were found. Instead, fine precipitates of 20-250 microns were present. Microscopically, there was a small number of cholesterol monohydrate crystals and bilirubin granules among an abundant amount of granular-crystalline material that was not morphologically cholesterol monohydrate crystals. The chemical composition of the precipitates (n = 4) was determined. There was a small amount of cholesterol (1.7% +/- 0.8%) and bilirubin (13.9% +/- 0.74%). The major component of the precipitate was a residue. On further analysis using thin-layer chromatography, high-performance liquid chromatography, and electron microprobe analysis, the residue was identified as a calcium salt of ceftriaxone. The residue also had identical crystal morphology and chromatographic elution profile as authentic calcium-ceftriaxone standards. It is concluded that ceftriaxone, after excretion and being concentrated in the gallbladder bile, can form a precipitate. The major constituent has been identified as a ceftriaxone-calcium salt.

Bile↗
PubMed1989

Tissue and ultrastructural localization of 5-hydroxytryptamine (serotonin) in the tissues of Ascaris suum with energy dispersive X-ray spectrometry of immunoreactive structures.

Muscle, hypodermis and gastrointestinal epithelial cells from adult female Ascaris lumbricoides var. suum were found to contain serotonin based upon glyoxylic acid induced histofluorescence and indirect immunolabeling with an antiserotonin monoclonal antibody conjugated to protein A-colloidal gold. Histofluorescence indicated that muscle-hypodermis and intestinal epithelial cells contained significant concentrations of 5-hydroxytryptamine while fluorescence was absent in the nerve cord and cuticle. Immunolabeling at the ultrastructural level indicated that serotonin was sequestered in electron-opaque patches, dense vesicles and mitochondria of the muscle-hypodermis and intestinal tissue. Perfusion of whole worms and isolated tissues with 10(4) M-serotonin further indicated: (1) immunolabeled patches and dense vesicles were often associated with cytoskeletal elements, (2) serotonin did not appear to enter the intestinal or muscle cells by endocytosis, (3) immunolabeled patches examined with energy dispersive X-ray spectrometry (X-ray microanalysis) were found to contain iron at concentrations approximately double that of the surrounding cytoplasm.

Animals↗
PubMed1993

Detection of human papillomavirus DNA in CaSki and HeLa cells by fluorescent in situ hybridization. Analysis by flow cytometry and confocal laser scanning microscopy.

CaSki and HeLa cell lines, isolated from human uterine carcinomas and containing integrated human papillomavirus (HPV) DNA type 16 and 18, respectively were used to evaluate the sensitivity of HPV-DNA detection on suspended cells by fluorescent in situ hybridization using flow cytometry and on corresponding cell deposits using confocal laser scanning microscopy (CLSM). HPV DNAs were detected in cell suspensions with biotinylated DNA probes and revealed with a three-step technique: a rabbit antibiotin antibody, a biotinylated goat anti-rabbit antibody and a streptavidin-fluorescein isothiocyanate complex. By flow cytometry, HPV DNA was detectable only in CaSki cells which contained about 600 copies of HPV DNA per cell. In HeLa cells, with only 20-50 copies of HPV DNA, flow cytometry could not detect HPV DNA, whereas CLSM permitted visualization of fluorescent labelling of HPV DNA hybrids. Furthermore, CLSM showed good preservation of cellular morphology and the nucleus was clearly recognizable after fluorescent in situ hybridization and counterstaining with propidium iodide. Moreover, this examination confirmed that the fluorescent foci were specifically confined to the cell nuclei.

DNA, Viral↗
PubMed2004

In vivo evaluation of resorbable bone graft substitutes in a rabbit tibial defect model.

Calcium sulfate as a bone graft substitute is rapidly resorbed in vivo releasing calcium ions but fails to provide long-term three-dimensional framework to support osteoconduction. The setting properties of calcium sulfate however allow it to be applied in a slurry form making it easier to handle and apply in different situations. This study examines the in vivo response of calcium sulfate alone and as a carrier for a coralline hydroxyapatite in an established bilateral corticocancellous defect model in rabbits. Defects were filled flush to the anterior cortex with a resorbable porous ceramic alone and in combination with calcium sulfate slurry, calcium sulfate slurry alone or calcium sulfate pellets and examined at time points up to 52 weeks. Specimens where assessed using Faxitron X-ray, light and electron microscopy. Calcium sulfate in either slurry or pellet form does indeed support new bone formation alone however, complete filling of the bone defect is not observed. Calcium sulfate in slurry form does however improve the surgical handling of particulate bone graft substitutes such as Pro Osteon 200 R, which remained as an osteoconductive scaffold for up to 52 weeks and may have played an important role in the ultimate closure of the cortical windows.

Absorbable Implants↗
PubMed2005

Effect of toothbrushing on titanium surface: an approach to understanding surface properties of brushed titanium.

OBJECTIVES: This in vitro study aimed to characterize the surface morphology and composition of tooth-brushed titanium casting and thereby to elucidate interactions between the metal and abrasive material in dentifrice. METHODS: Specimens were cast from CP Ti ingots and then mirror-finished. Two fluoride-free toothpastes containing crystalline CaHPO4.2H2O and amorphous SiO2 particles as abrasive were slurried with distilled water (15 g/30 mL). While toothbrushes were reciprocated at 120 strokes/min for 350,400 strokes, the specimens were brushed with the respective slurries under a load of 2.45 N. The brushed and non-brushed surfaces were characterized by means of SPM, EPMA, and XPS. SPM data were analyzed using one-way ANOVA, followed by post hoc Tukey test (p<0.01). RESULTS: Irrespective of toothpastes, toothbrushing had a significant influence on surface roughness. The CaHPO4.2H2O-containing paste produced much rougher surface than the SiO2-containing paste. Both the surfaces were chemically altered due to reactions with the respective abrasive materials. Abrading chips had dimensions of micron to submicron order. A number of chips were attached to abrasive particles. SIGNIFICANCE: The alterations of surface morphology and composition may affect biological responses of titanium in the oral environment. Dentifrice with lower abrasivity might be advisable for daily oral hygiene practice of patients with dental titanium devices.

Calcium Phosphates↗
PubMed2005

Characterization of Portland cement for use as a dental restorative material.

OBJECTIVES: The aim of this study was to evaluate the suitability of fast-setting cement formulations based on Portland cement as dental core build-up materials using two different methods of testing compressive strength and evaluation of setting times. METHODS: Four fast-setting cements based on Portland cement and their four respective densified with small particle (DSP) mortars were tested for setting time, constitution of cement by EDAX, and compressive strength using International and British Standards. Ordinary Portland cement (OPC) was used as a control. RESULTS: All the fast-setting cements had a similar elemental composition to OPC and the setting times were less than 7 min. The compressive strength of OPC was different between the two methods (P<0.001). All the fast-setting cements tested showed no difference in compressive strength regardless of the method of testing at 1 and 7 days (P>0.05), but the cylinders showed a lower compressive strength at 28 days (P<0.05). The OPC DSP mortar showed poorer compressive strength than OPC (P<0.01) at all times for cube testing but not for cylinder testing, where no difference was observed. The fast-setting DSP mortars had a lower compressive strength at 1 day (P<0.005) with both methods. At later times, there was no difference between the cements and DSP mortars for the cubes. SIGNIFICANCE: The pure fast-setting cements set in <7 min and were not susceptible to changes in the compressive strength testing procedure at 1 and 7 days but at 28 days all the fast-setting cements had a significantly higher strength with the test using cubes (P<0.05). A reduction in strength was observed at 28 days in cylinder testing. Most of the cements tested did not show encouraging strengths, however, one of the prototype cements tested could be a prospective dental restorative material.

Aluminum Oxide↗
PubMed2005

Evidence of the involvement of plant ligno-cellulosic structure in the sequestration of Pb: an X-ray spectroscopy-based analysis.

European walnut (Juglans regia) plants were grown in pots, on peat soil contaminated with lead (Pb), for four years. European walnut was chosen because it grows in Mediterranean climates, it yields a high biomass, and a fine quality wood. In the above ground parts Pb concentration was 1000 times lower than in roots: in 50 g roots there was 450 mg of Pb. Microanalysis of roots found in periderm more than 50% of the total root Pb. Pb L(III) EXAFS spectroscopy was performed on: root powder from Pb-exposed plants, Pb-impregnated cellulose and lignin. Comparison of plant material with lignin and cellulose helped to envisage a plant disposal strategy for Pb. This may consist in establishing links with large organic molecules, which are abundant constituents of cell walls. EXAFS spectroscopy evidenced the presence of Pb-O bindings within the ligno-cellulosic structure in roots. Lead was scantly conveyed to the shoots, giving to walnut plants an added asset in Pb phytostabilization.

Calcium↗
PubMed1994

Particles found in lung and brain following subureteral injection of polytetrafluoroethylene paste are not teflon particles.

We injected 0.3 ml. of polytetrafluoroethylene paste (Polytef) behind the left submucosal ureter in 4 mini-pigs and 4 dogs, and 2 dogs and 2 mini-pigs underwent sham operation and acted as controls. Two mini-pigs were injected with 0.5 ml. polytetrafluoroethylene (Teflon) suspended in 50 ml. of normal saline into a peripheral vein and 2 dogs were injected with 0.5 ml. polytetrafluoroethylene into a bladder vein. In addition, 4 dogs were injected with 0.1 ml. polytetrafluoroethylene paste suspended in 20 ml. saline into the right carotid artery. The lungs and brain from half of the animals who had subureteral and intravascular injection of polytetrafluoroethylene paste as well as sham operated animals were dissolved in sodium hypochlorite solution. The resulting organ suspensions were then centrifuged and the smear preparations of the precipitate were examined by polarized light microscopy, scanning electron microscopy and x-ray microanalysis. Polytetrafluoroethylene paste suspended in saline acted as positive control for polytetrafluoroethylene particles. Lungs and brain from the remaining animals were fixed in formalin solution. The brain and lungs of animals who underwent subureteral injection with a minimal amount of polytetrafluoroethylene paste carefully placed in the submucosal plane showed no evidence of polytetrafluoroethylene on histological examination, polarized light microscopy, scanning electron microscopy and x-ray microanalysis.

Animals↗
PubMed1983

Mulberry particles formed by red blood cells in human weddelite stones.

In a series of electron microscopic studies of human urinary stones, mulberry shaped particles were observed in 5 out of 51 calcium oxalate dihydrate (COD) containing stones. The mulberries were crenation products of red blood cells which adhered to the surface of bipyramid COD crystals, followed by impregnation of the COD. The red cells appeared to participate in the crystal growth by creating foci of surface nucleation, or by formation of a layer of crust on the surface of the bipyramids which caused deposition of an additional layer of calcium oxalate. The lipid containing organic matrix seems to have a role in COD crystal growth.

Calcium↗
PubMed1996

Histochemical analysis of dental hard tissues following bleaching.

The effect of commonly used bleaching materials on the dental hard tissues was tested in extracted human premolars. In each tooth, the apical two-thirds of the root was removed, and the remaining tooth stump was cut longitudinally into two equal segments. The segments were cleaned, dried, and divided into six experimental groups. Each group was treated with one of the following bleaching materials: 30% hydrogen peroxide (HP), 10% carbamide peroxide (CP), sodium perborate (SP), Nu-Smile (NS), Opalescence (Op), and DentlBright (DB). Treatment consisted of immersing the specimens in the respective test material followed by incubation at 37 degrees C for 7 days. The levels of calcium, phosphorus, sulfur, and potassium were measured in the enamel, dentin, and cementum. In the enamel, a significant reduction in the calcium/phosphorus (Ca/P) ratio was found following treatment with HP. In the dentin, a significant reduction in Ca/P ratio was found following treatment with HP,CP, DB, and Op. In the cementum, a significant reduction in the Ca/P ratio was found following treatment with HP, CP, NS, and Op. Changes in sulfur and potassium levels also occurred, but were usually not statistically significant. Significant reduction in sulfur levels occurred only in the cementum following treatment with CP and SP. Sulfur levels increased significantly following treatment with NS. Significant reduction in potassium levels occurred only in the dentin following treatment with CP. It is concluded that bleaching materials may adversely affect the dental hard tissues and should be used with caution.

Borates↗
PubMed2002

Titanium casting into phosphate bonded investment with zirconite.

OBJECTIVE: Reaction layer structure of the titanium castings surface was investigated when a new centrifuge-vacuum-pressure cast titanium machine and a zirconite inner investment was used. METHODS: The micro-hardness of titanium castings was measured using a Knoop's hardness tester. The structure of reaction layers and the distribution of elements on the surfaces of titanium castings were analysed by SEM and EDS, respectively, when zirconite was used as the inner investment for cast titanium. RESULTS: Knoop's micro-hardness at distances of 50 and 75 microm from the titanium casting surfaces made directly in phosphate investment molds had significantly harder surfaces than those made in zirconite coating molds. The thickness of the reaction layer on the surface of titanium castings was <35 microm with zirconite coating and was >50 microm without zirconite coating. The structure of reaction layer was divided into four layers from the outer surface to the inner including: sintering layer, alpha-Case, Si-rich layer, acicular crystal structure. The distribution of elements was different on each layer. SIGNIFICANCE: The thickness of the cast titanium reaction layer was reduced when the wax patterns were coated with zirconite. The surface of the titanium castings was smooth and without cracks, and the investment on the surface of titanium castings was easily removed.

Dental Casting Investment↗
PubMed2003

In vitro corrosion behavior of bioceramic, metallic, and bioceramic-metallic coated stainless steel dental implants.

OBJECTIVES: The most common metals and alloys used in dentistry may be exposed to a process of corrosion in vivo that make them cytotoxic. The biocompatibility of dental alloys is primarily related to their corrosion behavior. The aim of this work was to evaluate the corrosion behavior and thus the biocompatibility of the uncoated and coated stainless steels and compare the effect of type of coatings on corrosion behavior. METHODS: Three types of coatings, hydroxyapatite (HA), titanium (Ti), and a double-layer HA/Ti on AISI 316L stainless steel were made. HA coating was produced using plasma-spraying technique and Ti coating was made using physical vapor deposition process. In order to perform a novel double-layer composite coating, a top layer of HA was plasma-sprayed over a physical vapor deposited Ti layer on AISI 316L stainless steel. Structural characterization techniques including XRD, SEM and EDX were used to investigate the microstructure, morphology and crystallinity of the coatings. Electrochemical potentiodynamic tests were performed in physiological solutions in order to determine and compare the corrosion behavior of the coated and uncoated specimens as an indication of biocompatibility. RESULTS: Double-layer HA/Ti coating on AISI 316L SS had a positive effect on improvement of corrosion behavior. The decrease in corrosion current densities was significant for these coated specimens and was much lower than the values obtained for uncoated and single HA coated specimens. Ti coating on AISI 316L SS also has a beneficial effect on corrosion behavior. The results were compared with the results of corrosion behavior of HA coated commercially pure titanium (cpTi) and uncoated cpTi. SIGNIFICANCE: These results demonstrated that the double-layer HA/Ti coated 316L SS can be used as an endodontic implant and two goals including improvement of corrosion resistance and bone osteointegration can be obtained simultaneously.

Biocompatible Materials↗
PubMed1998

Electrodeposition of radioactive rhenium onto stents to prevent restenosis.

Radioactive stents are currently being evaluated for preventing restenosis. A major difficulty to overcome is the need to load any pre-manufactured stents with defined amounts of radioactivity at the time of use. Using stents that are preloaded by the manufacturer is not ideal because the stent length usually differs from the length needed for a specific lesion and the amounts of radioactivity varies widely due to ongoing decay of the source. Thus, we have developed a novel method that allows any currently used stainless steel or tantalum stent to be coated with radioactive rhenium. The method involves placing the stent in a series of rinsing and electroplating solutions, one containing radioactive rhenium (186Re, 188Re, or both). The overall processing time is 15 min and the procedure may be conveniently applied just prior to the stent insertion. The plated stent contains radioactive rhenium in a 1.2 microm-thick cobalt layer, with an outer 2 microm layer of gold. The gold layer gives the radioactive stent excellent radiochemical stability, good bending and biocompatibility properties, and improves stent visibility during fluoroscopy.

Angioplasty, Balloon, Coronary↗
PubMed1996

Distinct immuno-localization of mucin and other biliary proteins in human cholesterol gallstones.

BACKGROUND/AIMS: Cholesterol gallstones consist of cholesterol crystals and smaller amounts of pigments and calcium salts, arrayed on a mucin plus protein matrix. The localization of the various biliary proteins in the stones has not been characterized. We aimed to localize several biliary proteins in gallstones in order to determine their possible role in stone formation and growth. METHODS: The distribution of several matrix proteins and their relationships to the minerals were determined using immunostaining and EDAX microanalysis on hemisected cholesterol gallstones. RESULTS: Pigment areas were rich in calcium and contained Cu, P and S. These elements were absent in cholesterol regions. Mucin was identified in a three-dimensional network intercalated between cholesterol crystals and as septa between deposits of pigments and cholesterol; APF/CBP and ApN coated only the pigment deposits. No specific topographical localization was found for albumin or IgA. CONCLUSIONS: This suggests a role for mucin, APF/ CBP and ApN in the formation of cholesterol gallstones. We propose that cholesterol crystals bind directly to mucin, whereas calcium salts and pigments deposit on APF/CBP and ApN bind to the mucin.

Bile↗
PubMed1987

Argyria: the intradermal "photograph," a manifestation of passive photosensitivity.

X-ray microprobe and electron microscopic study was made of the remarkable blue-black pigmentation that sunlight elicits in patients with argyria. The patient under study had developed argyria following injection of silver nitrate as a sclerosant into his varicose veins 41 years ago. Similarities are demonstrated between the darkening of the skin and the darkening of a photographic film following light exposure. In both instances, colorless silver salts and compounds present in an inert matrix (collagen versus gelatin) are reduced by incident light to black metallic silver. This passive photosensitivity reaction leads to silver tattooing of the light-exposed skin and to photographic imaging in the film.

Aged↗