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PubMedDate not supplied

[Quantitative determination of lipoprotein-X in blood serum].

Lipoprotein X is a reliable indicator of cholestasis. The present paper describes a simple method for direct measurement of lipoprotein X. Low-density lipoproteins, very low density lipoproteins, chylomicrons and abnormal lipoprotein X are precipitated from the serum by treatment with a solution of polyethylene glycol (PEG) MW 6000. Then phosphotungstic acid and PEG 6000 solution are added to the precipitate. The lipoprotein X is in the supernatant, where its level can be estimated from the content of phospholipids and cholesterol. This method permits the determination of lipoprotein X in several samples simultaneously, requires no sophisticated equipment, is sensitive, reproducible, accurate, and cheap.

Adult↗
PubMed1979

Ultrastructure of an excitatory synapse.

The ultrastructure of the afferent synapse in hair cells of the lateral line-canal organ was studied using different fixation and staining techniques. Glutaraldehyde-fixed tissue without post-osmication, contrasted by section staining with uranyl acetate and lead citrate, was compared with (a) osmium tetroxide-fixed tissue followed by the same staining procedure, and with (b) glutaraldehyde-fixed tissue, block-impregnated with phosphotungstic acid (PTA). The results reveal a pronounced heterogeneity in the composition of the synaptic body, reflecting regional differences it the "intracleft substance", the synaptic structure defined by the PTA staining technique, is actually due to the glutaraldehyde fixation procedure and is apparently the outer leaflet of the postsynaptic membrane. A special technique that allows alternate sections of a series to be differentially stained for electron microscopy is proposed.

Animals↗
PubMed1997

Asynchronous changes in size, number, and shape of synapses in the developing superior colliculus.

The ultra-structural development of synapses in retino-receptive layers of the opossum superior colliculus was studied by the ethanolic phosphotungstic acid (E-PTA) method. There was a tendency for a slight reduction in the diameter of synaptic disks, a rise and fall of numerical densities and, except for an ephemeral period, a general increase in the proportion of "frown" among curve synapses. The lack of strict synchrony and the occurrence of different patterns of changes suggest that multiple factors contribute to synaptic maturation.

Animals↗
PubMed1990

The selective counterstaining of the plasma membrane improves the immunoelectron microscopic detection of neu protein at the cell border.

The distribution of the neu protein has been studied with two monoclonal antibodies in a post-embedding immunogold method on glycolmethacrylate sections. Labeling of the cytoplasm was due to binding to mitochondrial cristae. The detection of label at the plasma membrane was optimized by selectively staining the plasma membrane with phosphotungstic acid at low pH. Strong labeling was observed in breast carcinoma cells and faint labeling in other tissues. Brush borders were also found to be reactive. neu protein can thus be localized to baso-lateral and apical cell membranes. An important homology exists with a mitochondrial protein.

Breast↗
PubMed1994

Ultrastructural characterization of endogenous retroviral particles isolated from normal human placentas.

Human placental endogenous retroviral (ERV) particle isolates were investigated by ultrastructural evaluation. Although retrovirus-like structures in normal human placental tissue sections have been described, the precise nature of these particles has not previously been defined. By direct electron microscopic (EM) observation of placental ERV isolates that display retroviral reverse transcriptase (RTase) activity and a buoyant density consistent with type-C retroviruses (1.17 g/ml on sucrose), we have shown these to contain particles with characteristic retroviral ultrastructural features. Samples were stained with 1.0% phosphotungstic acid (PTA), 0.5% uranyl acetate (UA), and low-angle-shadowed with platinum/palladium. Isolated placental ERV particles have an apparent diameter of approximately 120 nm, are membrane-bound with a short surface fringe, and contain capsid particles (about 90 nm in diameter) within an internal matrix structure. These observations support the view that human placental cells normally express ERV particles.

Centrifugation, Density Gradient↗
PubMed1976

Electron microscopy of interphase chromosomes in situ; binding of permanganate to chicken erythrocytes.

From quantitative electron-microscope observations on the binding of permanganate to regions of erythrocytes and reticulocytes of known chemical composition, it is concluded that KMnO4, like phosphotungstic acid (PTA), binds preferentially to sites on proteins. Compared with PTA, KMnO4 binding exhibits less anomalous behaviour. The data support the hypothesis previously put forward that the 2 regions, or phases, in condensed chromatin differ in both molecular composition and concentration. The increase in binding to protein which occurs during nuclear haemolysis is interpreted in terms of protein-protein interaction in the chromatin of the intact cell.

Animals↗
PubMed2005

Malignant stromal tumor, so called "gastrointestinal stromal tumor", with rhabdomyomatous differentiation occurring in the gallbladder.

Gastrointestinal stromal tumors (GISTs) constitute the largest category of primary nonepithelial neoplasms of the gastrointestinal tract. It is extremely rare that this tumor occurs in the bile tract, and only a few cases have been reported. Immunohistochemically, the tumor cells revealed a phenotype similar to Cajal cells, occasionally with differentiation to smooth muscle cells or neural cells. We present a case of malignant stromal tumor similar to GISTs with rhabdomyomatous differentiation of the gallbladder in a 68-year-old woman. The resected tumor was predominantly composed of spindle cells with rhabdomyomatous differentiation. Immunohistochemical study revealed diffuse staining of tumor cells using vimentin despite negative staining for desmin or S-100. This indicated a mesenchymal origin of the cells without smooth muscle or neuronal differentiation. Myoglobin-positive cells, in which phosphotungstic acid hematoxylin staining revealed cross striations of the cytoplasm, suggested rhabdomyomatous differentiation. Diffuse positivity for KIT in the cells suggested that the pathogenesis of this tumor may resemble that of GIST. The tumor may have derived from a mesenchymal stem cell that had undergone partial rhabdomyomatous differentiation.

Aged↗
PubMed1995

Fine structure of tooth germs during the formation of enameloid matrix in Tilapia nilotica, a teleost fish.

Tooth germs were examined by light and transmission electron microscopy. Collagen fibrils were relatively dispersed and thin at the early and middle stages of formation of the enameloid matrix, when the enameloid layer was thin. At the late stage, the fibrils became thicker, reaching nearly 30 nm dia, and formed the interwoven thick bundles that are characteristic of teleost cap enameloid. Abundant flocculent and/or fine, network-like material, probably representing glycosaminoglycans or proteoglycans, was located between the collagen fibrils. Tall, columnar, inner dental epithelial cells contained abundant rough endoplasmic reticulum and many mitochondria, and a well-developed Golgi apparatus was seen around the nuclei at the late stage. Elongated vesicles enclosing fine, filamentous material that resembled procollagen granules, and large granules containing fibril-like structures that were 150 nm in thickness and had periodic cross-banding at 32-nm intervals, were usually observed near the Golgi apparatus. The contents of the large granules were well stained with phosphotungstic acid, which suggests that inner dental epithelial cells synthesize collagen fibrils. At this time, odontoblasts also contained abundant rough endoplasmic reticulum and mitochondria, a well-developed Golgi, several kinds of granule including those that probably contained procollagen, and many microtubules. It is proposed that odontoblasts are involved in the formation of a considerable portion of the enameloid matrix, including collagen fibrils.

Amelogenesis↗
PubMed1982

Electron density distribution of dry avian tendon.

The electron density profile along the unit cell of dry native collagen is determined. The method used is a modification of one that has been used successfully for wet native collagen. Uranyl acetate and phosphotungstic acid were used for isomorphous addition in phase determination and were located by electron microscopy. Structure factor magnitudes for dry collagen with and without the heavy metals were obtained from X-ray diffraction data. The first 9 orders were investigated. Standard Argand diagrams provided a unique solution for the phase of each. The resultant profile suggests that significant conformation changes occur in both the non-helical ends and the triple-helix portions of the molecules.

Animals↗
PubMed1992

Scanning force microscopy and cryo-electron microscopy of tobacco mosaic virus as a test specimen.

In this study, tobacco mosaic virus (TMV) provides a resolution criterion for specimen preparation methods as well as for imaging parameters of the scanning force microscope (SFM). We present scanning force microscopic images of the virus embedded in 0.5% buffered phosphotungstic acid solution adsorbed on a freshly cleaved mica surface, and imaged under atmospheric conditions. Individual TMV particles were clearly identified with a characteristic shape of long rods of about 300 nm long and 60-70 nm in apparent width due to the geometric parameters of the tip. The structure of the virus was compared with cryo-electron microscopic data of vitrified suspensions observed to a resolution of 1.15 nm. Uncoated TMV particles were also deposited on evaporated titanium thin films and imaged by SFM.

Cryopreservation↗
PubMed1995

Failure to demonstrate a protein coat on enamel crystallites by morphological means.

Phosphotungstic acid (PTA) treatment of section of Epon-embedded enamel dissolves the crystallites and stains material postulated to be crystal-bound proteins. Alternative, capillarity forces within the channels left after crystallite removal may draw in PTA. This prediction was tested on three systems. (1) Protein free synthetic hydroxyapatite was embedded in Epon; treatment of thin sections with PTA removed most crystals, leaving empty holes outlined by stain that could not represent protein. (2) Sections of rat incisor enamel were treated with PTA and then re-embedded in Epon and sectioned at 90 degrees to the original plane. In these sections-of-section the cut ends of dissolved crystallite profiles were coated with stain. To determine if stained protein coats can be detected in the absence of the crystallite profiles, Epon sections were partially demineralized with formic acid, re-embedded in Epon and sections-of section were PTA treated. Previously extracted crystallites left no stained coats, and only the crystallites that were not removed by formic acid left PTA-stained outlines. (3) PTA-treated sections of dogfish shark enameloid were flooded with 5-nm colloidal gold particles and sections-of-section were prepared. The presence of gold particles on the section surface and in holes previously occupied by crystallites suggested that PTA solution could also be sucked into similar holes. It is concluded that PTA outlines are not crystal-bound proteins but artefacts caused by stain lining holes left in the section when the crystallites have been extracted.

Animals↗
PubMed1992

Immunohistochemical and ultrastructural studies of intraluminal crystalloids in human prostatic carcinomas.

Intraluminal crystalloids (ICr) observed in 19 cases of incidental or invasive human prostatic carcinoma (PCa) and in a case of benign prostatic hyperplasia were examined extensively by immunohistochemistry and electron microscopy. They were brilliantly eosinophilic with haematoxylin and eosin, manifesting needle-like, triangular, rectangular, hexagonal and irregular lump-like in shape. They were strongly positive, dark blue, with phosphotungstic acid -haematoxylin (PTAH) stain in all cases examined. Among the human antibodies tested, epithelial membrane antigen (EMA) gave specifically positive immunostainability with ICr in all cases. Annual ring-like lamellar or concentric structures were detected by electron microscopy. Positive staining of ICr with PTAH and anti-EMA antibody is very useful as a diagnostic marker for PCa in human prostatic tissues.

Aged↗
PubMed1985

An improved EPTA staining method for synapses in rat cerebral cortex cultures.

Ethanolic phosphotungstic acid (EPTA) staining has proved to be a useful tool in electron microscopic studies on synapse formation and maturation in the developing brain. However, the available staining procedures did not give satisfactory contrast between synaptic structures and background in reaggregated cerebral cortex tissue grown in vitro. Such contrast is a prerequisite for reliable quantitative studies. We therefore varied several steps of the staining procedure and have succeeded in obtaining a modification which gives the required contrast. A short rinsing time after EPTA staining turned out especially to be very crucial.

Animals↗
PubMed2004

Malignant astrocytoma with binucleated granular cells in a Sprague-Dawley rat.

A 2-year-old Sprague-Dawley rat with hindlimb paralysis was diagnosed with a cerebral malignant astrocytoma. The distinctive feature of this astrocytoma was the presence of scattered binucleated cells that contained hypereosinophilic, 1-2 micro m in diameter, cytoplasmic granules. The neoplastic astrocytes stained positively for vimentin (VIM), lysozyme, and phosphotungstic acid hematoxylin (PTAH). Within the binucleated cells, granules stained with PTAH and periodic acid-Schiff (PAS) before and after diastase digestion. Ultrastructurally, neoplastic astrocytes were characterized by cytoplasmic aggregates of electron-dense intermediate filaments consistent with VIM and desmin. The cytoplasm of binucleated cells contained numerous phagolysosomes enlarged by myelin figures and glycoprotein or glycolipid. Intermediate filaments were not present. This is the first description, in the rat, of a neoplasm with features resembling the human granular cell astrocytoma. Our findings suggest that an astrocytic origin should be considered for the binucleated cells in this neoplasm.

Animals↗
PubMed1980

Morphological features of developing synapses as revealed by the E-PTA staining technique.

Synaptogenesis was studied in the optic tectum of chick embryos from the stage 40 to the hatching (last week of incubation). Our object was to characterize the morphological changes occurring in pre- and postsynaptic elements during this period of the development. Specialized interneuronal contact zones have been examined after conventional fixation procedure, or after staining of glutaraldehydefixed blocks with ethanolic phosphotungstic acid (E-PTA). Some mature synapses are seen also at the 14th incubation day (stage 40), and they increase in number to the hatching. In addition, immature synapses are seen in which presynaptic elements seem to be the less differentiated. Also the study of the evolution of the PTA-stainable paramembranous densities at the synaptic sites confirms these observations. These findings suggest a considerable prenatal synaptogenesis, as already demonstrated in various brain areas of the chick embryo, and also that the maturation of the postsynaptic site may precede and determine that of the presynaptic contact and vesicle accumulation.

Animals↗
PubMed1987

[Structural and functional reorganization of the interneuronal contacts of the cerebral cortex after a single convulsive paroxysm].

Using the technique of contrasting the cerebral tissue with phosphotungstic acid, the authors studied the structural and functional status of interneuronal contacts of the molecular layer of the sensomotor cortex in the brain of Krushinsky-Molodkina rats following convulsive sound stimulation and the subsequent audiogenic convulsive paroxysm. Marked reduction in the general number of synapses 4 h after the attack was attended by transformation of some flat functionally mature contacts into concave ones, which reflects the activation of the synaptic pool. The relative levels of concave and flat mature contacts returned to the initial level 8 to 24 h later.

Acoustic Stimulation↗
PubMedDate not supplied

[Effect of the isolation method and staining procedure on the morphology of the virion ribonucleoprotein of Sendai virus].

Electron microscopic examinations of morphology of Sendai virus ribonucleoprotein (RNP) isolated from purified virions by two methods and simultaneous staining of the preparations with uranyl acetate and phosphotungstate acid (PTA) were carried out. The staining method was shown not to influence the kind of nucleoproteins which had been isolated from disrupted virions by sucrose density gradient centrifugation. With both strains RNP appeared as strongly helixed filaments. The staining method, however, strongly influenced morphology of RNP isolated from disrupted virions by equilibrium centrifugation in cesium chloride density gradient. After uranyl acetate staining RNP had an appearance of hard filaments whereas in the same preparation stained with PTA partially and completely unwound spirals of nucleocapsid were found alongside with strongly spiralized structures.

Centrifugation, Density Gradient↗
PubMed1984

Electron microscopic observation on the developing rabbit and human vitreous collagen fibrils by negative staining.

The vitreous fibrils of developing eyes of 3-day-old rabbits and 24th week human fetuses were examined by the negative staining method (phosphotungstic acid) electron microscopically. Fibrils about 10 nm in diameter and with no periodic striated pattern as well as adult ones were observed. Peculiar fibers were observed in both vitreous samples preserved at low temperature. Particularly, dimeric segment-long-spacing (SLS) aggregates and fibrous-long-spacing (FLS)-like aggregates were observed in 3-day-old rabbit vitreous. FLS-like aggregates showed a 125-nm light band and 50-nm dark band periodic pattern and seemed to be formed with 300-nm overlapping of dimeric SLS aggregates. The present results indicate that the developing vitreous could possibly form various types of collagen fiber under native conditions.

Adult↗