Ultrastructure of Liver in Lactosyl Ceramidosis

Author(s):  
Z. Hruban ◽  
J. R. Esterly ◽  
G. Dawson ◽  
A. O. Stein

Samples of a surgical liver biopsy from a patient with lactosyl ceramidosis were fixed in paraformaldehyde and postfixed in osmium tetroxide. Hepatocytes (Figs. 1, 2) contained 0.4 to 2.1 μ inclusions (LCI) limited by a single membrane containing lucid matrix and short segments of curved, lamellated and circular membranous material (Fig. 3). Numerous LCI in large connective tissue cells were up to 11 μ in diameter (Fig. 2). Heterogeneous dense bodies (“lysosomes”) were few and irregularly distributed. Rough cisternae were dilated and contained smooth vesicles and surface invaginations. Close contact with mitochondria was rare. Stacks were small and rare. Vesicular rough reticulum and glycogen rosettes were abundant. Smooth vesicular reticulum was moderately abundant. Mitochondria were round with few cristae and rare matrical granules. Golgi complex was seen rarely (Fig. 1). Microbodies with marginal plates were usual. Multivesicular bodies were very rare. Neutral lipid was rare. Nucleoli were small and perichromatin granules were large. Small bile canaliculi had few microvilli (Fig. 1).

1974 ◽  
Vol 63 (1) ◽  
pp. 99-108 ◽  
Author(s):  
William B. Winborn ◽  
Leonard L. Seelig

Parietal cells of the stomach of the hamster show extensive amounts of dense material in a variety of organelles after prolonged exposure to a solution of osmium tetroxide. Conspicuous amounts of reduced osmium compounds are evident within the granular endoplasmic reticulum, perinuclear cisterna, and vesicular elements of the Golgi complex. Dense material is also apparent within cristae of the mitochondria, the surface coat of the microvilli of the intracellular canaliculus, and vesicular elements of the multivesicular bodies. Multivesicular bodies, containing numerous small osmiophilic elements, are often seen surrounding and/or in close contact with mitochondria. The proximity of the multivesicular bodies to the mitochondria appears to be related to an autophagic process involving degradation of mitochondria. The distribution and intensity of the precipitates within the organelles of the parietal cells vary in different regions of the gastric glands. The findings of this study emphasize that cell structures other than the Golgi complex may contain large concentrations of reduced osmium compounds after prolonged exposure to a solution of osmium tetroxide.


Author(s):  
C. N. Sun ◽  
H. J. White

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.


1978 ◽  
Vol 24 (10) ◽  
pp. 1173-1181 ◽  
Author(s):  
Nicholas Colotelo

The exudates or liquid droplets on various structures of a number of fungi were examined. The droplets were enveloped in membranous material and were associated with actively growing mycelia, including fruiting structures. Osmium tetroxide vapour-fixed droplets of Claviceps purpurea, Myrothecium roridum, Sclerotinia sclerotiorum, Sclerotium rolfsii, and Thanathephorus cucumeris did not dry to a powder but remained intact as spheres when freeze-dried. Fractured spheres, examined with the scanning electron microscope, showed the presence of a membranous structure similar to that of rapidly frozen colloidal solutions with the ice crystals removed by sublimation. Locules or cavities within the freeze-dried droplets are thought to be due to the entrapment of air when droplets coalesce. Biochemical analyses of the exudates showed that acid phosphatase, β-glucosidase, acid and alkaline protease, RNase polygalacturonase and cellulase enzymes as well as oxalic acid and ammonia were present.


1966 ◽  
Vol 31 (2) ◽  
pp. 319-347 ◽  
Author(s):  
Robert E. Smith ◽  
Marilyn G. Farquhar

The nature and content of lytic bodies and the localization of acid phosphatase (AcPase) activity were investigated in mammotrophic hormone-producing cells (MT) from rat anterior pituitary glands. MT were examined from lactating rats in which secretion of MTH1 was high and from postlactating rats in which MTH secretion was suppressed by removing the suckling young. MT from lactating animals contained abundant stacks of rough-surfaced ER, a large Golgi complex with many forming secretory granules, and a few lytic bodies, primarily multivesicular bodies and dense bodies. MT from postlactating animals, sacrificed at selected intervals up to 96 hr after separation from their suckling young, showed (a) progressive involution of the protein synthetic apparatus with sequestration of ER and ribosomes in autophagic vacuoles, and (b) incorporation of secretory granules into multivesicular and dense bodies. The content of mature granules typically was incorporated into dense bodies whereas that of immature granules found its way preferentially into multivesicular bodies. The secretory granules and cytoplasmic constituents segregated within lytic bodies were progressively degraded over a period of 24 to 72 hr to yield a common residual body, the vacuolated dense body. In MT from lactating animals, AcPase reaction product was found in lytic bodies, and in several other sites not usually considered to be lysosomal in nature, i.e., inner Golgi cisterna and associated vesicles, and around most of the immature, and some of the mature secretory granules. In MT from postlactating animals, AcPase was concentrated in lytic bodies; reaction product and incorporated secretory granules were frequently recognizable within the same multivesicular or dense body which could therefore be identified as "autolysosomes" connected with the digestion of endogenous materials. Several possible explanations for the occurrence of AcPase in nonlysosomal sites are discussed. From the findings it is concluded that, in secretory cells, lysosomes function in the regulation of the secretory process by providing a mechanism which takes care of overproduction of secretory products.


1965 ◽  
Vol 26 (1) ◽  
pp. 99-123 ◽  
Author(s):  
Milton W. Brightman

From 10 minutes to 3½ hours after the intraventricular injection into rats of 15 to 100 mg of ferritin, an appreciable fraction of the protein, visualized electron microscopically, traverses the ependymal epithelium by diffusing along the dense intercellular substance of the luminal open junction and thence, by circumventing discrete intercellular fusions which partition rather than seal the interspace. These partitions shunt additional protein into the cell, where ferritin is transported within pinocytotic vesicles to the lateral and basal plasma-lemma and, presumably, back into the interspace again. The basal interspace is irregularly distended by pools of moderately dense "filler" within which ferritin accumulates. The larger fraction of protein enters the ependyma by pinocytosis and is eventually segregated within membrane-enclosed organelles such as vacuoles, multivesicular bodies, and dense bodies, where the molecules may assume a crystalline packing. As a result of the accumulation of ferritin within these inclusions and within filler substance, only a small amount of protein remains to enter the underlying parenchyma. Presentation of ferritin to prefixed cells leads to a random dispersion of free cytoplasmic ferritin. This artifactual distribution in both prefixed and postfixed cells is concurrent with disruption of cell membranes.


1968 ◽  
Vol 37 (1) ◽  
pp. 89-104 ◽  
Author(s):  
Nancy J. Lane

The neuronal perikarya of the grasshopper contain sudanophilic lipochondria which exhibit an affinity for vital dyes. These lipochondria are membrane-delimited and display acid phosphatase activity; hence they correspond to lysosomes. Unlike those of most vertebrates, these lysosomes also hydrolyze thiamine pyrophosphate and adenosine triphosphate. Like vertebrate lysosomal "dense bodies," they are electron-opaque and contain granular, vesicular, or lamellar material. Along with several types of smaller dense bodies, they are found in close spatial association with the Golgi apparatus. The Golgi complexes are frequently arranged in concentric configurations within which these dense bodies lie. Some of the smaller dense bodies often lie close to or in association with the periphery of dense multivesicular bodies. Further, bodies occur that display gradations in structure between these multivesicular bodies and the dense lysosomes. Acid phosphatase activity is present in the small as well as the larger dense bodies, in the multivesicular bodies, and in some of the Golgi saccules, associated vesicles, and fenestrated membranes; thiamine pyrophosphatase is found in both the dense bodies and parts of the Golgi complex. The close spatial association of these organelles, together with their enzymatic similarities, suggests the existence of a functional or developmental relationship between them.


1965 ◽  
Vol 25 (3) ◽  
pp. 545-562 ◽  
Author(s):  
Daniel Szollosi

Electron microscope observations of osmium tetroxide-fixed rat eggs indicate that small nucleoli are extruded from pronuclei in a sharply demarcated time period after sperm penetration. Approximately 4½ hours after sperm penetration, fine fibrous material aggregated in distinct loci along the inner surface of the nuclear envelope and condensed into small, dense bodies. The term tertiary nucleolus or extrusion body is used to designate the forming bodies. The small tertiary nucleoli form distinct protrusions from the pronuclei during the following developmental period and finally bud off into the cytoplasm, carrying with them a small portion of the double nuclear envelope. The extrusion bodies can be observed only in the vicinity of the pronuclei and have not been seen near the cell membrane. The fate of the tertiary nucleoli is not known; apparently they transform or disappear after they have passed into the cytoplasm. Eleven hours after sperm penetration, tertiary nucleoli are not present near the nuclear membrane and the extrusion activity has apparently ceased. Large and small nucleoli react similarly to cytochemical reagents: they are Feulgen negative; they are positive to the Millon, Sakaguchi, brom-phenol blue, and PAS reactions. Azure B stain combined with nuclease extraction indicates the presence of small amounts of RNA in the nucleoli.


2013 ◽  
Vol 583 ◽  
pp. 137-144
Author(s):  
Cosmin Alexandru Buzila ◽  
Iulian Antoniac ◽  
Florin Miculescu ◽  
Marius Dumitrescu ◽  
Ionel Droc

A 55-year-old female who undergone mitral valve replacement eight years ago with a mechanical graft, presented accusing sudden decrease of effort tolerance and two episodes of pulmonary edema in the last month. Anamnesis, physical examination, electrocardiogram (ECG), transthoracic echocardiography and coronarography were performed. The mechanical valve leaflets and the tissue surrounding the prosthetic ring were evaluated by: optical microscopy (hematoxylin eosin stain and immunohistochemistry), scanning electron microscopy (SEM) and EDAX analysis (Energy Dispersive X-ray spectroscopy). Anamnesis: inferior myocardial infarction in 2006, congestive cardiac insufficiency, pulmonary hypertension, and arterial hypertension. Clinical examination and ECG: minimal perimaleolar edema bilateral, sinus rhythm on admission. Transthoracic echocardiography: 55% ejection fraction, a pression gradient across the mitral valve (Gmax/Gmed= 24/11 mmHg), tricuspid regurgitation, and pulmonary hypertension. A mitral prosthetic valve’s leaflet was found immobile. No thrombus was evidenced. Coronarography: an immobile graft’s leaflet, stenoses on the right coronary artery, stenosis on left anterior descending artery and occlusion at the circumflex artery emergence. The prosthesis was replaced, and two coronary artery bypasses were performed. Macroscopic examination: a fibroconjunctive tissue expansion in close contact with the leaflet. Histopathological evaluation: muscle cells with altered phenotypes, fibroblasts along with fibrous connective tissue and calcium depots areas. SEM evaluation: tissue depots on the immobile leaflet, suggesting that the connective tissue expansion was blocking the leaflet’s movements. EDAX analysis: the metallic leaflets surface was made of tantalum; sodium, calcium and chloride deposits were also detected. Graft failure was caused by the tissue proliferation affecting leaflet’s mobility. SEM is a viable method for failed cardiovascular grafts investigation.


Author(s):  
Kuen-Shan Hung ◽  
M. Sue Hertweck ◽  
John D. Hardy ◽  
Clayton G. Loosli

A small number of fibroblasts (alveolar septal cells, pulmonary interstitial cells, mesenchymal cells) are present in the alveolar areas of the adult lungs. These cells, like fibroblasts elsewhere, may be important in production and maintenance of connective tissue fibers found throughout the lung. Very little attention has been given to these cells in studies of the lungs. Therefore their precise structure and function are not clear. This report demonstrates filaments in the fibroblasts in the alveolar ducts and alveolar walls of the mouse lungs.The lungs were inflated with 2% glutaraldehyde, post-fixed with 1% osmium tetroxide and processed for electron microscopy. The cell body and processes of the alveolar fibroblast are located in the interalveolar septum (Fig. I).


Author(s):  
Kevin M. Imel ◽  
J. Franklin Bailey ◽  
Mark DeSantis

Damage to axons in the neonatal mammal results in the death of the affected neurons rather than survival and axonal regeneration as generally seen in the adult. This phenomenon has been well documented using indirect methods such as counting cells in histological sections of the experimentally damaged versus control tissue. Recent experiments have shown that cells with osmiophilic dense bodies, which have been termed “granules”, become apparent at the light microscopic level in ipsilateral lumbar dorsal root ganglia after destruction of the sciatic nerve on that side in neonatal rats. Within 84 - 96 hours after the lesion, the rat pups were anesthetized, perfused with 4% paraformaldehyde and the ganglia post-fixed in 1% unbuffered osmium tetroxide for at least 72 hours. Such a lengthy exposure to osmium was found to be necessary to visualize the granules.This offers the possibility of using these granules as a direct indicator of neuronal death in the lumbar dorsal root ganglia. The cells containing these granules were usually chromatolytic neuronal somata which had an eccentrically positioned nucleus (when a nucleus or nuclear remnant was still present) and peripheral aggregation of the rough endoplasmic reticulum (i.e. Nissl bodies).


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