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Oncocytic carcinoma arising in submandibular gland with immunohistochemical observations and review of the literature.

We report a case of oncocytic carcinoma arising in submandibular gland. The tumour occurred in the left submandibular gland of an 82-year-old Japanese man. Histologically, the tumour was mostly composed of large cells with eosinophilic granules in the cytoplasm and they were arranged in the solid sheets, islands with duct-like structure and cords. The tumour cells had aggressively invaded muscles and perineural tissues, and cervical lymphatic metastasis was frequently observed. Histochemically, the tumour cells were strongly positive for phosphotungstic acid-hematoxylin (PTAH) stain, and we diagnosed this malignant tumour as oncocytic carcinoma. Immunohistochemically, the tumour cells reacted positively for cytokeratin 7, 8, 19, epithelial membrane antigen (EMA), alpha-1-antichymotrypsin and carcinoembryonic antigen (CEA), but negative for cytokeratin 13, 14, smooth muscle actin (HHF35) and S-100 protein (S-100). Tumour was diagnosed as oncocytic carcinoma in submandibular gland. Its characteristics are discussed in term of its histopathological and immunohistochemical features.

Adenocarcinoma↗

Neuronal structure of the lacertilian parietal eye, I: A retrograde label and electron-microscopic study of the ganglion cells in the photoreceptor layer.

The cellular connectivity of the lacertilian parietal eye is not well understood. Because the intercellular connections establish the foundation for information processing, we have investigated cellular connectivity of one cell type in this simple vertebrate retina. We also developed an in vitro preparation to study the anatomy of the parietal eye visual system. Horseradish peroxidase transport in the in vitro preparation revealed a class of displaced ganglion cells occupying positions among the photoreceptors, in a location where the presence of interneurons had been suggested. Three-dimensional reconstruction at the electron-microscopic level showed that the morphology and synaptic input of these displaced ganglion cells is different from that of the previously known ganglion cells. The displaced ganglion cells receive an average of about 13 ribbon synapses from photoreceptors. The ribbon input is equally distributed between the soma and dendritic arbor. Junctional membrane measurement and ethanolic phosphotungstic acid-staining provided evidence for the existence of non-ribbon synaptic contacts (synaptoid junctions). Displaced ganglion cells make about 20 synaptoid junctions, 65% of which are on the dendritic arbor. The morphology of the displaced ganglion cell is such that a significant measure of synaptic input to the dendritic arbor will be transmitted to the soma.

Animals↗

Aqueous speciation studies of europium(III) phosphotungstate.

The incorporation of lanthanide ions into polyoxometalates may be a unique approach to generate new luminescent, magnetic, and catalytic functional materials. To realize these new applications of lanthanide polyoxometalates, it is imperative to understand the solution speciation chemistry and its impact on solid-state materials. In this study we find that the aqueous speciation of europium(III) and the trivacant polyoxometalate, PW9O34 9-, is a function of pH, countercation, and stoichiometry. For example, at low pH, the lacunary (PW11O39)7- predominates and the 1:1 Eu(PW11O39)4-, 2, forms. As the pH is increased, the 1:2 complex, Eu(PW11O39)2 11- species, 3, and (NH4)22[(Eu2PW10O38)4(W3O8(H2O)2(OH)4].44H2O, a Eu8 hydroxo/oxo cluster, 1, form. Countercations modulate this effect; large countercations, such as K+ and Cs+, promote the formation of species 3 and 1. Addition of Al(III) as a counterion results in low pH and formation of [Eu(H2O)3(alpha-2-P2W17O61)]2, 4, with Al(III) counterions bound to terminal W-O bonds. The four species observed in these speciation studies have been isolated, crystallized, and characterized by X-ray crystallography, solution multinuclear NMR spectroscopy, and other appropriate tech-niques. These species are 1, (NH4)22[(Eu2PW10O38)4(W3O8(H2O)2(OH)4].44H2O (P; a=20.2000(0), b=22.6951(6), c=25.3200(7) A; alpha=65.6760(10), beta=88.5240(10), gamma=86.0369(10) degrees; V=10550.0(5) A3; Z=2), 2, Al(H3O)[Eu(H2O)2PW11O34].20H2O (P, a=11.4280(23), b=11.5930(23), c=19.754(4) A; alpha=103.66(3), beta=95.29(3), gamma=102.31(3) degrees; V =2456.4(9) A3; Z=2), 3, Cs11Eu(PW11O34)2.28H2O (P; a=12.8663(14), b=19.8235(22), c=21.7060(23) A; alpha=114.57(0), beta=91.86(0), gamma=102.91(0) degrees ; V=4858.3(9) A3; Z=2), 4, Al2(H3O)8[Eu(H2O)3(alpha-2-P2W17O61)]2.29H2O (P; a=12.649(6), b=16.230(8), c=21.518(9) A; alpha=111.223(16), beta=94.182(18), gamma=107.581(17) degrees ; V=3842(3) A3; Z=1).

Crystallography, X-Ray↗

Poly(alkylcyanoacrylate) nanocapsules: physicochemical characterization and mechanism of formation.

Nanocapsules of poly(isobutylcyanoacrylate) and poly(isohexylcyanoacrylate) were prepared by addition of the monomer to an organic phase and subsequent mixing of the organic phase to an aqueous phase containing poloxamer 188, 238 or 407. Gel permeation chromatography indicated that in contrast to literature reports, polymerization occurred in the organic phase and nanocapsules were obtained by interfacial precipitation of the polymer without any significant change of the molecular weight. Addition of SO2 to the organic phase before the introduction of the monomer allowed preparation of nanocapsules with a lower molecular weight. Nanospheres were prepared in a similar way albeit using an organic phase that was completely miscible within the aqueous phase so that solid spheres were obtained. Density gradient centrifugation revealed that nanocapsules had a density intermediate between nanospheres and an emulsion prepared in the same way without addition of monomer to the organic phase. Further, the process used to prepare nanocapsules had a high yield since no oil droplets or nanospheres were obtained by this process. Zeta potential of the nanocapsules and spheres was found to be related to the molecular weight of the polymer: values as high as approximately -42 mV were obtained for low molecular weight nanocapsules (MW approximately 1000) compared to approximately -10mV for the emulsion and the high molecular weight nanocapsules (MW approximately 100,000). Surface charge of the nanocapsules and molecular weight of their polymeric wall conditioned the adsorption capacity of poloxamers. Moreover, the highest adsorption was measured with the most hydrophobic poloxamer. These observations agree with previous work conducted on hydrophobic surfaces.

Adsorption↗

Eight prion strains have PrP(Sc) molecules with different conformations.

Variations in prions, which cause different incubation times and deposition patterns of the prion protein isoform called PrP(Sc), are often referred to as 'strains'. We report here a highly sensitive, conformation-dependent immunoassay that discriminates PrP(Sc) molecules among eight different prion strains propagated in Syrian hamsters. This immunoassay quantifies PrP isoforms by simultaneously following antibody binding to the denatured and native forms of a protein. In a plot of the ratio of antibody binding to denatured/native PrP graphed as a function of the concentration of PrP(Sc), each strain occupies a unique position, indicative of a particular PrP(Sc) conformation. This conclusion is supported by a unique pattern of equilibrium unfolding of PrP(Sc) found with each strain. Our findings indicate that each of the eight prion strains has a PrP(Sc) molecule with a unique conformation and, in accordance with earlier results, indicate the biological properties of prion strains are 'enciphered' in the conformation of PrP(Sc) and that the variation in incubation times is related to the relative protease sensitivity of PrP(Sc) in each strain.

Animals↗

A novel sample holder allowing atomic force microscopy on transmission electron microscopy specimen grids: repetitive, direct correlation between AFM and TEM images.

A novel sample holder that allows atomic force microscopy (AFM) to be performed on transmission electron microscope (TEM) grids is described. Consequently, AFM and TEM images were repeatedly obtained on exactly the same sample area. For both techniques, a thin carbon film was used as the imaging substrate. Although these techniques have been previously used in conjunction, AFM and TEM images on exactly the same area have not been repeatedly obtained for any system. Correlation of AFM and TEM images is useful for work where the three-dimensional topographical information provided by the AFM could be used to better interpret the two-dimensional images provided by the TEM and vice versa. To demonstrate the applicability of such correlation, new results pertaining to a fibrillar collagen system are summarized.

Fibrillar Collagens↗

Electron microscopy of negatively stained and unstained fibrinogen.

Electron microscope images of negatively stained fibrinogen are predominantly asymmetric rods 450 A in length and about 60 A in width. The molecules appear to have considerable flexibility, and mass distribution along the major axis is not uniquely distinguished despite apparent beading in some particles. Scanning transmission electron microscopy of unstained fibrinogen again demonstrates that a majority of molecules are rodlike. The results differ from those obtained by negative staining in that a substantial fraction of images are trinodular with striking resemblance to those obtained by C. E. Hall and H. S. Slayter [J. Biophys. Biochem. Cytol. (1959) 5, 11--16] using the mica replica technique. The above results were obtained on glow-discharged carbon substrate films by a simple low-concentration, long-attachment-time modification of standard deposition methods that is diffusion controlled and depends on concentration and time but is independent of pH, buffer, and other staining conditions. Evidence is presented that standard attachment procedures result in artifactual images. Any models of fibrinogen in solution consequently must encompass properties that permit its visualization as an asymmetetric rod by electron microscopy as first suggested by Hall and Slayter 20 years ago.

Animals↗

Ultrastructural identification of protein bodies, cellular markers of human catecholamine neurons, in a temporal lobe ganglioglioma.

A temporal lobe ganglioglioma, surgically removed from an 8-year-old body, and a human brainstem at the level of locus coeruleus (LC) were processed for light microscopy (LM), with formalin fixation and paraffin embedding, and for electron microscopy (EM) with glutaraldehyde fixation, potassium permanganate postfixation, phosphotungstic acid-hematoxylin block-staining, and epoxy resin embedding. The paraffin sections were stained with toluidine blue O/rhodamine B and observed under epi-fluorescence. The thin sections for EM were viewed directly without further staining. The neuronal neoplastic cells of ganglioglioma and the neurons of LC are known to produce catecholamines. Both also contain spherical protein bodies (pb), cellular markers that identify catecholamine neurons in humans. The ultrastructural characteristics of the pb in LC were compared with those of the pb in neoplastic ganglion cells. These bodies had an identical ultrastructure, in both tissues, consisting of electron-lucent core surrounded by an electron-dense thin rim. The rhodamine B-stained sections also emphasized the identical morphology of the pb in ganglioglioma and LC. Based on the EM comparison, these brightly fluorescing spherical bodies are ideal markers for identifying in LM, the clusters of large neoplastic cells, representing neurons, which are the most important clue to the correct diagnosis of gangliogliomas.

Biomarkers↗

2-Butoxyethanol female-rat model of hemolysis and disseminated thrombosis: X-ray characterization of osteonecrosis and growth-plate suppression.

We recently proposed a chemically induced rat model for human hemolytic disorders associated with thrombosis. The objective of the present investigation was to apply a noninvasive, high-magnification X-ray analysis, the Faxitron radiography system, to characterize the protracted bone damage associated with this 2-butoxyethanol model and to validate it by histopathology. Groups of female Fischer 344 rats were given 0, 250, or 300 mg of 2-butoxyethanol/kg body weight daily for 4 consecutive days. Groups were then sacrificed 2 hours or 26 days after the final treatment. The treated animals displayed a darkened purple-red discoloration on the distal tail. Histopathological evaluation, including phosphotungstic acid-hematoxylin staining of animals sacrificed 2 hours after the final treatment, revealed disseminated thrombosis and infarction in multiple organs, including bones. The Faxitron MX-20 specimen radiography system was used to image selected bones of rats sacrificed 26 days posttreatment. Premature thinning of the growth plate occurred in the calcaneus, lumbar and coccygeal vertebrae, femur, and ilium of the treated animals. Areas of decreased radiographic densities were seen in the diaphysis of the femur of all treated animals. The bones were then examined histologically and showed a range of changes, including loss or damage to growth plates and necrosis of cortical bone. No thrombi were seen in the animals sacrificed at 30 days, but bone and growth plate changes consistent with prior ischemia were noted. The Faxitron proved to be an excellent noninvasive tool that can be used in future studies with this animal model to examine treatment modalities for the chronic effects of human thrombotic disorders.

Animals↗

An electron microscope study of the epithelium in the normal mature and immature mouse cornea.

1. An electron microscope study at high resolution of the corneal epithelium of the normal mature and immature mouse revealed new information regarding the submicroscopic appearance of these cells. 2. Two thin dense lines separated by a less dense area constituted the structure of the limiting surface membrane of epithelial cells; the thickness of this membrane was about 80 A. 3. Some differences in the appearance of the cytoplasm and mitochondria of cells from the immature mouse cornea and the appearance of the cytoplasm and mitochondria of cells from the adult mouse cornea were observed. 4. The basement membrane appeared as a dense band about 600 A wide separating the basal epithelial cells from the substantia propria. Suggestions of periodicity were seen in some phosphotungstic acid-treated specimens. 5. Round bodies believed to be bacteria were seen on the surface of the outer epithelial cells in the adult mouse cornea but not in the immature, unopened eye.

Animals↗

Fine structure of lipid-depleted mitochondria.

The fine structure of mitochondria and submitochondrial vesicles depleted of their lipid by extraction with aqueous acetone was studied. Thin sections of mitochondrial membranes depleted of more than 95% of their lipid retained the unit membrane structure. Densitometer tracings of the electron micrographs showed that the unit membrane of extracted mitochondria was, on the average, wider than that of unextracted controls and showed a greater variation in width. The outer membrane was lost in mitochondria from which 80-95% of the lipids was extracted. Inner membrane particles were present on submitochondrial vesicles depleted of up to 85% of their lipids. However, when more than 95% of the lipid was removed, few, if any, particles remained attached to the membranes but many particles were found unattached in the background. When lipid was restored to lipid-deficient preparations, the mitochondrial membranes were found to be devoid of inner membrane particles but were fully active with respect to succinate-cytochrome c reductase activity.

Animals↗