scholarly journals Elemental analysis of valvular and atherosclerotic calcification

2021 ◽  
Vol 10 (3) ◽  
pp. 26-33
Author(s):  
L. A. Bogdanov ◽  
N. Yu. Osyaev ◽  
Yu. D. Bogdanova ◽  
R. A. Mukhamadiyarov ◽  
A. R. Shabaev ◽  
...  
Keyword(s):  
Elemental Analysis ◽  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
Mineral Deposit ◽  
Mineral Deposits ◽  
Atherosclerotic Plaques ◽  
Aortic Valves ◽  
Nucleation Sites ◽  
Initial Nucleation ◽  
Backscattered Scanning Electron Microscopy

Aim. To analyze the topographic patterns of valvular and atherosclerotic calcification growth.Methods.           Dysfunctional aortic valves (n = 18) and atherosclerotic plaques (n = 20) were fixed in formalin, postfixed in 1% osmium tetroxide, consecutively stained by 2% osmium tetroxide and 2% uranyl acetate, and embedded into epoxy resin (Epon) with the further grinding and polishing ofthe samples. Upon the counterstaining by lead citrate and sputter coating with carbon, samples were visualized by backscattered scanning electron microscopy. Elemental analysis was conducted via energy-dispersive X-ray spectroscopy. Measurement of Ca/P ratio within the mineral deposits was carried out employing a pool table principle (i.e., in the center of the deposit, in the near and far circumferences (clockwise), and in control regions around the mineral deposit). Topographic patterns of calcifications were modeled using the correlation analysis.             Results. Significant correlation was revealed between the Ca/P ratio in the deposit center and in the near and far circumferences of deposit in both in valvular (r = 0,35-0,78 - near circumference; r = 0,63-0,69 - far circumference) and atherosclerotic mineral deposits (r = 0,37-0,56 - near circumference; r = 0,48-0,63 - far circumference), suggesting the hierarchical growth of cardiovascular calcification around the initial nucleation sites.Conclusion.       Valvular and atherosclerotic calcifications development is concentric.

scholarly journals Ultrastructural pathology of bioprosthetic heart valves with infectious endocarditis

2021 ◽  
Vol 6 (3) ◽  
pp. 25-34
Author(s):  
R. A. Mukhamadiyarov ◽  
I. V. Milto ◽  
A. G. Kutikhin
Keyword(s):  
Extracellular Matrix ◽  
Heart Valves ◽  
Osmium Tetroxide ◽  
Giant Cells ◽  
Uranyl Acetate ◽  
Diglycidyl Ether ◽  
Multinucleated Giant Cells ◽  
Bioprosthetic Heart Valves ◽  
Sputter Coating ◽  
Backscattered Scanning Electron Microscopy

Aim. To study the ultrastructure of mitral bioprosthetic heart valves (BHVs) which failed due to infective endocarditis.Materials and Methods. Here we examined 7 ethylene glycol diglycidyl ether-treated xenopericardial BHVs excised during repeated BHV replacement because of prosthetic endocarditis. After being fixed in formalin and postfixed in osmium tetroxide, BHVs were dehydrated and stained in uranyl acetate with the subsequent embedding into epoxy resin, grinding, polishing, and lead citrate counterstaining. Upon the sputter coating with carbon, we visualised the BHV microanatomy by means of backscattered scanning electron microscopy at 15 kV voltage.Results. The extracellular matrix underwent degradation and disintegration resulting in loosening, fragmentation, and reduction in the electron density of collagen and elastin fibers. We observed a number of recipient cells (macrophages, multinucleated giant cells, neutrophils, endothelial cells and smooth muscle cells) within the BHVs. The highest number of cells was localized on the valve surfaces. The localization of the recipient cells on the ventricular and atrial surfaces was different. The central part of the valves was abundantly populated by macrophages.Conclusion. Prosthetic endocarditis is accompanied by the migration of recipient cells into the BHV structure, which is the consequence of surface and extracellular matrix disintegration.

Cellular Contacts Within the Interhemal Membrane of the Rat Placenta at Term

1974 ◽  
Vol 32 ◽  
pp. 146-147
Author(s):  
William P. Jollie
Keyword(s):  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
Capillary Endothelium ◽  
En Bloc ◽  
Aldehyde Fixation ◽  
Chorioallantoic Placenta

By routine EM preparative techniques, the tissues which, collectively, separate maternal and fetal bloods in the fully formed chorioallantoic placenta of the rat have been shown to consist of three chorionic layers, or trophoblast, and a layer of allantoic capillary endothelium [Fig. 1]. Relationships between these layers are best demonstrated by special techniques, viz., cacodylate-buffered aldehyde fixation, collidine-buffered osmium tetroxide postfixation, and en bloc staining with uranyl acetate. By using this method on placentas at term, the cells of the outermost chorionic layer (Trophoblast 1) appear to be attached to each other by means of maculae adherentes which sometimes occur in clusters [Fig. 2].

Membranous alterations in the cytoplasm and nucleus in a primitive neuroectodermal tumor of the brain

1978 ◽  
Vol 36 (2) ◽  
pp. 628-629
Author(s):  
C. N. Sun ◽  
C. Araoz ◽  
H. J. White
Keyword(s):  
Nuclear Envelope ◽  
Tumor Cells ◽  
Osmium Tetroxide ◽  
Primitive Neuroectodermal Tumor ◽  
Uranyl Acetate ◽  
Neuroectodermal Tumor ◽  
Annulate Lamellae ◽  
Lead Citrate ◽  
Thin Sections ◽  
The Brain

The ultrastructure of a cerebral primitive neuroectodermal tumor has been reported previously. In the present case, we will present some unusual previously unreported membranous structures and alterations in the cytoplasm and nucleus of the tumor cells.Specimens were cut into small pieces about 1 mm3 and immediately fixed in 4% glutaraldehyde in phosphate buffer for two hours, then post-fixed in 1% buffered osmium tetroxide for one hour. After dehydration, tissues were embedded in Epon 812. Thin sections were stained with uranyl acetate and lead citrate.In the cytoplasm of the tumor cells, we found paired cisternae (Fig. 1) and annulate lamellae (Fig. 2) noting that the annulate lamellae were sometimes associated with the outer nuclear envelope (Fig. 3). These membranous structures have been reported in other tumor cells. In our case, mitochondrial to nuclear envelope fusions were often noted (Fig. 4). Although this phenomenon was reported in an oncocytoma, their frequency in the present study is quite striking.

Cytoplasmic Invaginations in Trophoblasts and Epithelial Cells of Rat

1971 ◽  
Vol 29 ◽  
pp. 524-525
Author(s):  
Iracema M. Baccarini
Keyword(s):  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
Nuclear Inclusions ◽  
Mucous Cells ◽  
Electron Microscopic ◽  
Cervical Epithelium ◽  
Intact Membrane ◽  
Microscopic Studies ◽  
Abnormal Cells ◽  
Nuclear Features

Some morphological nuclear features (invaginations) in normal and abnormal cells have been described in several electron microscopic studies. They have been referred to by others as blebs, loops, pockets, sheets, bodies, nuclear inclusions and cytoplasmic invaginations. Identical appearing structures were found in cells of the uterine cervical epithelium, in trophoblasts of blastocysts and in trophoblasts of rat placenta.Methods. Uterine cervix (normal rats), rat placenta (9-10 days gestation) and blastocyst were placed in 3% glutarahdehyde for 3 hours. The tissue was washed in phosphate buffer for 24 hours, postfixed in 1%. buffered osmium tetroxide for 1-2 hours and embedded in epon araldite. Sections were double stained with uranyl acetate and lead citrate and viewed in E. M. Siemens 200.Observations. Nuclear invaginations were found in basal, parabasal and mucous cells of the cervix epithelium, in trophoblasts of blastocyst and in trophoblasts of placenta. An oval, round or elongated invagination contained heterogenously cytoplasm surrounded by a double intact membrane; usually several invaginations were found in the same nucleus.

The Ultrastructure of Metastatic Human Mammary Carcinoma After Prolonged Estrogen Therapy

1967 ◽  
Vol 25 ◽  
pp. 180-181
Author(s):  
John H.L. Watson ◽  
John L. Swedo ◽  
R.W. Talley
Keyword(s):  
Mammary Carcinoma ◽  
Osmium Tetroxide ◽  
Estrogen Therapy ◽  
Ultrastructural Level ◽  
Uranyl Acetate ◽  
Human Mammary Carcinoma ◽  
Surgical Biopsy ◽  
Lead Hydroxide ◽  
Preliminary Study ◽  
Hormonal Therapies

A preliminary study of human mammary carcinoma on the ultrastructural level is reported for a metastatic, subcutaneous nodule, obtained as a surgical biopsy. The patient's tumor had responded favorably to a series of hormonal therapies, including androgens, estrogens, progestins, and corticoids for recurring nodules over eight years. The pertinent nodule was removed from the region of the gluteal maximus, two weeks following stilbestrol therapy. It was about 1.5 cms in diameter, and was located within the dermis. Pieces from it were fixed immediately in cold fixatives: phosphate buffered osmium tetroxide, glutaraldehyde, and paraformaldehyde. Embedment in each case was in Vestopal W. Contrasting was done with combinations of uranyl acetate and lead hydroxide.

Reactivity of Pulmonary Alveolar Cells to Crocidolite Asbestos Fibers

1982 ◽  
Vol 40 ◽  
pp. 334-335
Author(s):  
Charles L. Sanders ◽  
Roy R. Adee
Keyword(s):  
Osmium Tetroxide ◽  
Intratracheal Instillation ◽  
Uranyl Acetate ◽  
Fiber Suspension ◽  
Nacl Solution ◽  
Alveolar Cells ◽  
Asbestos Fibers ◽  
Fischer Rats ◽  
Mineral Silicates

Asbestos is a generic name for a group of hydrated mineral silicates that occur naturally in a fibrous form. The early interactions of asbestos fibers with alveolar cells in large part determines their long-term toxicity. Young adult, SPF, Fischer rats were given a single intratracheal instillation of 2 mg crocidolite asbestos suspended in 0.5 ml of 0.9% NaCl solution. About 80% of the fibers had lengths of less than 10 ym as measured on light micrographs of the fiber suspension. Two rats were killed at 3 hr, 1 d and 1, 4, 8, 12 and 16 wk after instillation and the lungs instilled with 8 ml McDowell - Trumps at 20 cm H2O. Lung tissue was dehydrated and sputtered coated with palladium-gold for SEM or post-fixed in osmium tetroxide, embedded in epoxy resin and sections stained with uranyl acetate and lead citrate for TEM.

Ultrastructural comparison of prolactin adenomas of pituitary in men and women

1988 ◽  
Vol 46 ◽  
pp. 346-347
Author(s):  
R.C. Caughey ◽  
U.P. Kalyan-Raman
Keyword(s):  
Endoplasmic Reticulum ◽  
Electron Microscope ◽  
Phosphate Buffer ◽  
Pituitary Adenomas ◽  
Osmium Tetroxide ◽  
Secretory Granules ◽  
Uranyl Acetate ◽  
En Bloc ◽  
Men And Women ◽  
Transmission Electron

Prolactin producing pituitary adenomas are ultrastructurally characterized by secretory granules varying in size (150-300nm), abundance of endoplasmic reticulum, and misplaced exocytosis. They are also subclassified as sparsely or densely granulated according to the amount of granules present. The hormone levels in men and women vary, being higher in men; so also the symptoms vary between both sexes. In order to understand this variation, we studied 21 prolactin producing pituitary adenomas by transmission electron microscope. This was out of a total of 80 pituitary adenomas. There were 6 men and 15 women in this group of 21 prolactinomas.All of the pituitary adenomas were fixed in 2.5% glutaraldehyde, rinsed in Millonig's phosphate buffer, and post fixed with 1% osmium tetroxide. They were then en bloc stained with 0.5% uranyl acetate, rinsed with Walpole's non-phosphate buffer, dehydrated with graded series of ethanols and embedded with Epon 812 epoxy resin.

Ultrastructure of myoepithelial cell in parotid gland

1988 ◽  
Vol 46 ◽  
pp. 342-343
Author(s):  
C. N. Sun
Keyword(s):  
Parotid Gland ◽  
Osmium Tetroxide ◽  
Individual Cell ◽  
Branching Processes ◽  
Muscle Cells ◽  
Myoepithelial Cells ◽  
Uranyl Acetate ◽  
Thin Sections ◽  
Gland Carcinoma ◽  
Parotid Gland Carcinoma

Myoepithelial cells have been observed in the prostate, harderian, apocrine, exocrine sweat and mammary glands. Such cells and their numerous branching processes form basket-like structures around the glandular acini. Their shapes are quite different from structures seen either in spindleshaped smooth muscle cells or skeletal muscle cells. These myoepithelial cells lie on the epithelial side of the basement membrane in the glands. This presentation describes the ultrastructure of such myoepithelial cells which have been found also in the parotid gland carcinoma from a 45-year old patient.Specimens were cut into small pieces about 1 mm3 and immediately fixed in 4 percent glutaraldehyde in phosphate buffer for two hours, then post-fixed in 1 percent buffered osmium tetroxide for 1 hour. After dehydration, tissues were embedded in Epon 812. Thin sections were stained with uranyl acetate and lead citrate. Ultrastructurally, the pattern of each individual cell showed wide variations.

Electron Microscopic Examination of a Transplantable Rat Mammary Carcinoma

1968 ◽  
Vol 26 ◽  
pp. 20-21
Author(s):  
S. Shirahama ◽  
G. C. Engle ◽  
R. M. Dutcher
Keyword(s):  
Electron Microscope ◽  
Osmium Tetroxide ◽  
Electron Microscopic Examination ◽  
Undifferentiated Carcinoma ◽  
Uranyl Acetate ◽  
Sprague Dawley ◽  
Electron Microscopic ◽  
North West ◽  
X Irradiation ◽  
Tissue Specimens

A transplantable carcinoma was established in North West Sprague Dawley (NWSD) rats by use of X-irradiation by Engle and Spencer. The tumor was passaged through 63 generations over a period of 32 months. The original tumor, an adenocarcinoma, changed into an undifferentiated carcinoma following the 19th transplant. The tumor grew well in NWSD rats of either sex at various ages. It was invariably fatal, causing death of the host within 15 to 35 days following transplantation.Tumor, thymus, spleen, and plasma from 7 rats receiving transplants of tumor at 3 to 9 weeks of age were examined with an electron microscope at intervals of 8, 15, 22 and 30 days after transplantation. Four normal control rats of the same age were also examined. The tissues were fixed in glutaraldehyde, postfixed in osmium tetroxide and embedded in Epon. The plasma was separated from heparanized blood and processed as previously described for the tissue specimens. Sections were stained with uranyl acetate followed by lead citrate and examined with an RCA EMU-3G electron microscope.

Banded Structures in the Connective Tissue of Meningioma

1976 ◽  
Vol 34 ◽  
pp. 234-235
Author(s):  
C. N. Sun ◽  
H. J. White
Keyword(s):  
Connective Tissue ◽  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
Collagen Fibrils ◽  
Lead Citrate ◽  
Connective Tissues ◽  
Native Collagen ◽  
Routine Procedures

Previously, we have reported on extracellular cross-striated banded structures in human connective tissues of a variety of organs (1). Since then, more material has been examined and other techniques applied. Recently, we studied a fibrocytic meningioma of the falx. After the specimen was fixed in 4% buffered glutaraldehyde and post-fixed in 1% buffered osmium tetroxide, other routine procedures were followed for embedding in Epon 812. Sections were stained with uranyl acetate and lead citrate. There were numerous cross striated banded structures in aggregated bundle forms found in the connecfive tissue of the tumor. The banded material has a periodicity of about 450 Å and where it assumes a filamentous arrangement, appears to be about 800 Å in diameter. In comparison with the vicinal native collagen fibrils, the banded material Is sometimes about twice the diameter of native collagen.

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