Scanning microscopy最新文献

We examined 15 variably-sized gallstones, taken from an elderly male, by backscattered electron imaging and energy-dispersive X-ray microanalysis to learn the structural and distribution patterns of gallstone calcium (Ca-) salts. Of the 13 cholesterol-rich stones, nine stones had peripheral concentric layers of Ca-carbonate, whereas 2 stones had peripheral layers of Ca-phosphate. No Ca-salts were detected from 2 cholesterol-rich stones. The 2 stones containing Ca-phosphate had no Ca-salt cores, whereas the stones containing Ca-carbonate were separated into 3 different types: two stones with a Ca-carbonate core, four stones with several Ca-bilirubinate cores of glass-like structure, and 3 stones lacking Ca-salt cores. A closer view of the Ca-salt layers, which may be occasionally coexistent with Ca-bilirubinate, mainly showed either laminate deposits or numerous globules with a few laminae. Of the 2 cholesterol-poor stones, one had dispersed particles mainly of Ca-phosphate, and the other had loosely dispersed particles with small amounts of Ca-phosphate, bilirubinate, and/or palmitate. Some relationship between the size and Ca-salt species of these gallstones was suggested. Gallstones collected from the same individual showed a considerable heterogeneity of Ca-salts.

We studied articular disks and endoarticular loose bodies taken from patients suffering from different types of temporomandibular joint (TMJ) pathology. The scanning electron microscopy (SEM) analysis of the disks and the endoarticular loose bodies was followed by a chemical-compositional analysis using an energy dispersive spectrometer (EDS) and by characterization of the crystalline phases by X-ray powder diffraction (XRD). The articular disks were composed of a central radiopaque area lacking any evident structural features, surrounded by compact bundles of collagen fibers. EDS and XRD analyses showed that endodiscal radio-opaque areas were hydroxyapatite. By SEM, we observed a fibrous network only in circumscribed areas of the endoarticular loose bodies. The chemical-compositional analysis showed that the loose bodies were composed of calcite (CaCO3). The results of this investigation, along with the clinical history of the patients, allow us to formulate some hypotheses regarding the etiopathogenesis of these structural anomalies. The endodiscal calcifications could be the result of a chronic inflammatory process that produces displastic alterations of the articular disk. Moreover, an acute inflammatory process with modifications in the mechanisms of the synovial fluid turnover seems to be the event that leads to the formation of endoarticular loose bodies.

The degree of supersaturation of saliva with calcium (Ca) is related to the mineral phase of enamel in erupted teeth, the incidence of caries, and the formation of calculus. The mechanisms for regulating salivary Ca concentration are therefore of relevance to dentistry. Sections of rabbit, rat and human submandibular gland (SMG) were processed for immuno-histochemistry with a specific anti-plasma membrane Ca-pump antibody, 5F10. Western blots confirm that the molecular weight of the proteins identified by our antibody (135 kDa) is consistent with an appropriate molecular weight for PMCA antigen (135-150 kDa). Tissue sections were also processed for in situ hybridization to study the distribution of the PMCA mRNA isoforms. In mammals, the PMCA1 gene is reported to code for a PMCA protein with a role in maintaining the intracellular Ca levels in both epithelial and non-epithelial cells. Other genes including the PMCA2 and PMCA4 genes may code for PMCA proteins specific to Ca transporting tissues. Our studies demonstrate cytoplasmic labeling of PMCA mRNA with hPMCA-1 and hPMCA-4 specific cDNA probes in humans, and rPMCA-1 and rPMCA-2 specific oligonucleotide probes in rats. Labeling of PMCA protein and all mRNA isoforms was found in the cytoplasm of the interlobular and intralobular ducts (except for intercalated ducts). The demonstrated presence of PMCA in SMGs of rabbit, rat, and man, may suggest a role for PMCA in the regulation of intracellular Ca and in a mechanism for regulating and maintaining the high concentration of Ca in salvia.

Organization of cytoskeleton and cell contacts were studied by immunochemistry and electron microscopy in confluent HT29 cultured cells following exposure to 0.5 and 1.0 Gy doses of X-ray. Microtubules were resistant to irradiation, whereas, the actin and intermediate filaments disrupted rapidly following the treatment and their components appeared as clumps of actin and cytokeratin aggregates in the cytoplasm as demonstrated by immunochemistry. Loss of cell contacts and decrease in the number of desmosomes was also characteristic of irradiated cells. Electron microscopy revealed intact desmosomes in control cells and abnormal desmosomes in the irradiated samples characterized by the absence of tonofilaments. The perinuclear filament network and cortical filaments were well detectable by electron microscopy. Under the effect of irradiation, the perinuclear filaments almost disappeared and, at the same time, small bundles of filaments were formed irregularly in the cytoplasm associated with amorphous material.

This study investigates tissue responses after laser irradiation of the rabbit ureter, which serves as an experimental model for rectourogenital fistulae of children. Twenty-five rabbit ureters were irradiated intraluminally by a Nd:YAG laser 1320 nm (2 Watt, 20 seconds and 3 Watt, 8 seconds) via an applicator with radialsymmetrical light distribution. Immediately, 2 weeks, 4 weeks, 8 weeks, and 16 weeks after irradiation, the ureters were X-rayed with contrast solution and prepared for light and transmission electron microscopy. For the parameters employed, no apparent morphological differences could be observed. Immediately, the central laser zone showed a transmural thermonecrosis prevailed by cellular destruction, condensed ground substance and occlusion of most vascular lumina. Peripheral laser zones displayed urothelial vacuolations. Between 2 and 16 weeks, urothelial regeneration and ingrowth of granulation tissue caused a luminal stenosis or occlusion followed by transformation into scar tissue. In some peripheral laser zones, a hydroureter with marked luminal dilatation developed. We conclude that the ureter is occluded if the expanding force of the growing scar tissue exceeds the hydrostatic pressure of the obstructed urine. A laser occlusion of rectourogenital fistulae will be easier to achieve since fistula occlusion does not entail an obstruction of the urine flow.

Disruption of cytoplasmic and spindle microtubules by colchicine or nocodazole increases mitotic index, but it also enhances apoptosis in isolated mouse thymocytes; the apoptotic index exceeds 20% after 4 hours of incubation with either drug (5% in controls). Apoptosis was confirmed by DNA fragmentation, and was blocked by calcium chelators and inhibitors of protein synthesis. The apoptotic effect of microtubule disrupting drugs (MDD) was directed to interphase thymocytes and was independent on MDD action on mitotic cells. However, cell death of mitotically arrested cells showed ultrastructural changes similar in many aspects to apoptosis.

5-Fluorouracil (5-FU) is a widely used antineoplastic agent. 5-FU induced cardiotoxicity is a still relatively unknown side-effect of this drug. This phenomenon could be due to a direct cytotoxic effect on the endothelial cells. We tested this hypothesis in an experimental study in rabbits, by scanning or transmission electron microscopic evaluation of endothelium in small arteries (the central artery of the ear) after in vivo treatment with 5-FU. Both local and systemic effects of 5-FU on endothelium were studied 15, 30, 60 and 120 minutes after intra-arterial or intraperitoneal treatment. Perfusion fixation at physiological pressure and temperature was used in order to minimize damage to the endothelium during the preparation procedure. Eighteen rabbits weighing 2.5-3.0 kg were used, and 6 animals served as controls. The following parameters were evaluated: vessel wall and endothelial cell contraction, cell edema, cytolysis, occurrence of denuded areas, platelet adhesion/aggregation and fibrin formation. For the description of each parameter a scale of negative points was used. Irreversible cell damage was observed in 5-FU treated animals: disruption of the endothelial sheet and patchy exposure of the subendothelium, sometimes as a focus for thrombus formation. Our findings support the hypothesis that the thrombogenic effect of 5-FU secondary to its direct cytotoxic effect on endothelium might be one of the pathophysiological mechanisms behind 5-FU induced cardiotoxicity.

This study investigated enamel laser conditioning as an alternative to acid etching in bracket therapy. In preliminary experiments optimal laser parameters for achieving a bond strength of 6-10 N/mm2 were defined. Enamel surface morphology was assessed and the ablation depth was measured on serial enamel sections. Thirty human molars were exposed to 193 nm ArF-excimer laser radiation (energy density: 260 mJ/cm2) by single pulse application of 23 nanoseconds. Thirty molars were etched with phosphoric acid (37%) for 60 seconds. The brackets from the treated molars and 30 untreated molars were debonded vertically for tensile bond strength measurement. Roughened enamel surfaces were attained by 450 and 900 laser pulses with a mean ablation depth of 10.13 +/- 4.84 microns. After 1-10 laser pulses, the enamel surface appeared intact. The tensile bond strength was 6.63 +/- 2.18 N/mm2 in the laser-treated group (1 pulse), 8.75 +/- 3.61 N/mm2 in the acid-etched group, and 4.61 +/- 3.15 N/mm2 in the untreated group. We conclude a laser-selective ablation of the membranous enamel pellicle. Since the irradiated area can be adapted to bracket base and the enamel surface remains morphologically intact, pulsed ArF-excimer laser treatment seems to be superior to the acid etching technique.

It is generally recognized that calcium oxalate crystal formation in urine is induced by heterogeneous nucleation. However, there is no consensus as to the nature of the nucleation substrate. Evidence is provided in this paper that membranous cellular degradation products are the most likely candidates because they: (1) are ubiquitous in urine and urinary stones; (2) are found in close association with crystal deposits in the kidneys; and (3) can induce nucleation of crystals from a metastable solution of calcium oxalate in vitro and metastable urine in vivo.

The marked radiosensitivity of renal tissue represents a limitation on the total radiotherapeutic dose that safely can be applied to treatment volumes that include the kidneys. Radiation nephropathy is characterized by a progressive reduction in renal hemodynamics associated with a severe anemia. The latter is often normochromic normocytic in character, but can progress to a microangiopathic hemolytic anemia. The pathogenic mechanisms responsible for the development of radiation nephropathy remain ill-defined. Experimental studies which allow serial determinations of functional, morphologic, and cell kinetic radiation-induced changes indicate that primarily glomerular but also tubular alterations occur in the primary stages of radiation nephropathy. Glomerular capillary endothelial cell loss is seen within several weeks of irradiation. Remaining endothelial cells exhibit increased permeability leading to a subendothelial transudate. Mesangiolysis also is observed. In contrast, podocytes appear to be relatively unaffected at this stage. The endothelial changes appear to resolve, but the mesangial lesions progress, with hypercellularity and/or hypertrophy, increased mesangial matrix, mesangial sclerosis, and ultimately, glomerulosclerosis. These mesangial changes are similar to those observed in other chronic glomerulopathies. Dietary protein restriction, corticosteroids, and ACE-inhibitors all can reduce the severity of experimental radiation nephropathy.