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Ultrastructure of adamantinoma of long bones.

Electron microscopic examination of three adamantinomas of long bones revealed structures usually observed in both mesenchymal and epithelial tumours. Tumour cells showing high alkaline phosphatase activity frequently tended to line clefts in connective tissue, resembling endothelial cells. The long processes of such cells showed fenestration. In areas showing fibre production the tumour cells were in close relationship to collagen fibrils. The latter were found, together with microfibrils, between the processes and above the basement membranes. The tumour cells were interconnected by desmosomes with tonofilaments and contained numerous bundles of microfilaments. All three cases revealed tiny intracytoplasmic inclusions resembling Weibel-Palade endothelial bodies. In addition, some of the structures in the lumena contained definite acid mucosubstances. A squamous cell pattern was present in only one of the three specimens. The coincidence of divergent structures in a single specimen has led us to the conclusion that the so-called adamantinoma of long bone might be possibly related to tumours of mesodermal or mesectodermal origin.

Alkaline Phosphatase↗

Chronic megakaryocytic-granulocytic myelosis--an electron microscopic study. I. Megakaryocytes and thrombocytes.

The fine structure of the bone marrow in chronic megakaryocytic-granulocytic myelosis (CMGM) was studied in 5 nontreated patients to investigate possible malignant proliferation of megakaryocytes and the role of megakaryopoiesis in fibrillogenesis, terminating in osteomyelofibrosis. In comparison with normal megakaryopoiesis there is an enormous increase of the megakaryocytic cell line and many immature and atypical forms are seen. Most conspicuous are microforms, nuclear-cytoplasmic disorganization and nuclear inclusions. Asynchrony of maturation causes abnormal thrombocytogenesis with premature detachment of platelets resulting in immature and peculiar giant forms of thrombocytes. Besides megakaryocytes appearing superficially normal the maturation anarchy of many cells is so severe that by analogy with observations in other leukaemic cells these abnormalities are thought to be representative of a malignant growth. Moreover, there is a striking accumulation of microfibrils and single collagen fibres around megakaryoblasts. Since these cells contain all those organelles commonly associated with fibre production the initial step for fibrillogenesis may therefore arise from the megakaryoblasts prior to platelet release, or any fibroblast proliferation.

Blood Platelets↗

Some morphological details of the endochondral layer of labyrinthine bone. A comparative anatomical light and transmission electron microscopit study.

The endochondral layer of labyrinthine bone of dog (Canis f. intermedius Woldrich) is examined by means of light and transmission electron microscopy. The osteocytes of the fine-fibred, alamellar "embryonic skein bone" (Meyer 1927) are surrounded by a fibre-less matrix. They correspond to osteocytes in long bones. Some of them are producing microfibrils according to formation of preosseous tissue which is called "intraosseous osteogenesis" (Knese 1970). Others are characterized by signs of micropetrosis or indicate osteocytic osteolysis. In the perivascular mesenchym of some vessels giant cells are visible which according to their ultrastructural characteristics are identified as osteoclasts. The occurrence of an embryonic type of bone in endochondral layer of otic capsule throughout life is discussed considering oxygenation via perivascular canalizition as well as biochemical aspects.

Animals↗

Fluorescence microscopical visualization of elastic fibres using basic fuchsin.

We show that fluorescence microscopy after staining of tissue sections with basic fuchsin (BF) can be used successfully for the demonstration of elastic fibres. Using double staining with BF and antibodies reacting with microfibrils of elastic fibres (anti-SAP) we showed that BF reacts with the elastin core of elastic fibres and the elastin poor terminal branches of the subepidermal elastic fibre system. Small amounts of bound BF were easily seen by fluorescence microscopy (FL) but not by ordinary light microscopy. Both frozen sections and sections of paraffin embedded tissues could be stained. The BF-FL staining procedure is simple to perform and, due to its selectivity, it may be useful for detecting elastic fibres in various tissues at the light microscopical level.

Elastic Tissue↗

Histochemical staining of lymphatic anchoring filaments.

Lymphatic anchoring filaments are stained by means of histochemical methods that demonstrate disulfide-groups. Thiosulfation of sections either followed by aldehyde-fuchsin staining or by Alcian Blue +0.8 M MgCl2 staining has been used. Lymphatic anchoring filaments display striking fine structural similarities to "elastic fiber microfibrils" and both kinds of fibers are characterized by disulfide content.

Animals↗

Electron microscopic studies on zonular fibers. II. Changes of the zonular fibers after the treatment with collagenase, alpha-chymotrypsin and hyaluronidase.

Adult rabbit zonular fibers maintained in their native condition were treated with collagenase, alpha-chymotrypsin and hyaluronidase, and were observed with the electron microscope. The results obtained were as follows: 1. Collagenase digested the lens capsule, but not the zonular fibers. 2. Long time collagenase action obscured the cell membrane of the lens epithelium and the basal lamina of the ciliary epithelium. 3. Washing with 0.9% NaCl increased the collagenase action on the lens capsule. 4. Alpha-chymotrypsin digested the zonular fibers and the zonular lemalla, but not the lens capsule and the basal lamina of the ciliary epithelium. 5, Hyaluronidase only slightly changed the lens capsule. 6. The vitreous fibers were digested by collagenase, but not by alpha-chymotrypsin or hyaluronidase. Thes results together with the review of recent literature indicate that the zonular fiber has a nature close to that of the microfibril of elastic fiber.

Animals↗

Electron microscopic localization of chitin using colloidal gold labelled with wheat germ agglutinin.

The lectin wheat germ agglutinin (WGA) has a binding site which is able to bind a sequence of three N-acetyl-glucosamine residues. Therefore, it has a very strong affinity for the polymers of this sugar, especially chitin. Colloidal gold can be labelled with WGA and used as a specific electron-dense marker for the electron-microscopic localization of chitin. The specificity of the WGA-gold binding can be checked by competitive inhibition with 5-10 mM triacetyl chitotriose. The reliability of this method was tested in three species. In the formation zone of the radula of the snail, Biomphalaria glabrata Say, chitin or chitin precursors were localized in vesicles of the odontoblasts, outside the extremely long microvilli of odontoblasts and in the newly formed teeth. The inner peritrophic envelope of the earwig, Forficula auricularia L., is characterized by an orthogonal texture of bundles of microfibrils that are thought to contain chitin. The presence of chitin was proved using the present method. In the peritrophic membranes of the blowfly, Calliphora erythrocephala Meigen, it was possible to differentiate between chitin and glycoproteins which have N-acetylglucosamine residues.

Animals↗

[Electron microscopic examination of the morphogenesis of the elastic fibre in elastosis senilis and pseudoxanthoma elasticum (author's transl)].

We undertook to investigate the elastic and collagenous fibers of senile elastosis (s.E.) and pseudoxanthoma elasticum (P.e.). This was done by comparing the above with normal skin under the electron microscope. The main difference is the increasing occurance of a fine granular-filamentous substance composed of microfibrils of varying thicknesses. A new formation arises as this substance is excreted by the activated fibroblasts and the combines extracellulary with the pathological elastic fibers. In the case of P.e. there is an additional calcification. It seems that with the elastogenesis of s.E. small elastic fibers are formed. P.e., in contrast, shows irregular electron-dense reticular sheets with numerous holes. The alteration of collagen observed in both diseases is considered as the primary change (which arises in both cases through the fine granular-filamentous material). In both diseases it is a matter of alterations of the little vessels, in endothel and basement membranes. We find here the same fine granular-filamentous substance in the elastic fibers of lamina interna, where as in P.e. we can see the calcification.

Adult↗

Ultrastructure of the contrasting types of keratinization seen in the tail epidermis of the laboratory mouse Mus musculus.

The mouse tail epidermis undergoes contrasting forms of keratinization. Around the hair follicle there is a granular layer containing keratohyalin granules, and nuclei are absent from the horny layer. In the scale regions keratohyalin is not formed and nuclear remnants are retained in the horny cells as in parakeratosis generally. These findings from light microscopy were confirmed by transmission electron microscopy. The complete breakdown of organelles in the follicular regions contrastred with the retention of effete nuclei in the scales. Some of these nuclear remnants were pyknotic as in abnormal human parakeratosis, but most were further degraded with loss of nuclear membranes. In the boundary zone between the follicular and scale regions the epidermal cells had a few small keratohyalin granules and also showed incomplete degradation of nuclear remnants in the horny cells. The change from living epidermal cells to dead keratinized cells was abrupt in both the follicular and scale regions. In both sites the plasma membranes of the horny cells were thickened and there was a cytoplasmic meshwork of microfibrils in the cells.

Animals↗

[Ultrastructure of endocardial lesions in the carcinoid syndrome (author's transl)].

In a 67-year-old female patient with metastasising carcinoid of the small intestine there were right-sided endocardial plaques characteristic of the carcinoid syndrome. Ultrastructurally, the lesions showed proliferation of smooth muscle cells and myofibroblasts, which were surrounded by basement-membrane-like material and occasionally showed intercellular connections of the macula adhaerens type. The ground substance consisted of acid mucopolysaccharides, normal collagen fibrils, and microfibrils. The endocardial plaques in the carcinoid syndrome differ morphologically from other endocardial lesions by the lack of amorphous elastin. The formation of plaques and its relation to the endocrine activity of the carcinoid tumors is discussed, and pertinent literature is reviewed.

Aged↗

Morphological variability of smooth muscle cells in human nasal swell bodies.

The complex functional behavior of nasal swell bodies is still not completely understood. In the present study the histology of the vessels involved in the swelling mechanism is examined and the ultrastructural appearances described of the different types of smooth muscle cells located in the vascular wall of swell bodies in the human inferior turbinate. Even though the majority of smooth muscle cells of the nasal swell bodies showed a normal, elongated appearance comparable to other smooth muscle cells elsewhere in the body, a variety of cells with atypical shapes could be detected that have not been described previously in vessels of the nasal mucosa. The diameters of the smooth muscle cells in general were strikingly variable. The individual smooth muscle cells were surrounded by a basal lamina that was occasionally disrupted or doubled. Myoblasts were separated by a connective tissue space containing collagen fibrils, mature elastin fibers and bundles of microfibrils. The latter two types of fibers and fibrils occurred mainly in the outer parts of the muscular coat. The endowment of cytoplasmic components was similar in all smooth muscle cells of the vascular wall in the swell bodies. These findings indicate that the specific feature of smooth musculature presumably resides in the unusual morphological variability of the single cells present, as well as in the striking heterogeneity of the arrangement of bundles of these cells in the vascular wall.

Connective Tissue↗

The evolution of experimental endarteritis in the rabbit abdominal aorta. Light and transmission electron microscopy.

Experimental aortic intimal thickening has been induced in rabbits by sheathing the vessel with a polyethylene cuff. The alterations have been examined by light and transmission electron microscopy, during 12 months. An irregular intimal thickening develops as soon as the 15th day and includes numerous myofibroblasts with some other cells of monocytic or endothelial type. Microfibrils, elastic aggregates and collagen fibers are found in the intercellular space. Simultaneously, the media undergoes a fragmentation of the elastic laminae and the adventitia shows a capillary angiectasis and a granuloma. After 3 months there is, between the intimal smooth muscle cells, a progressive increase of elastic and collagenous material. In the media, elastic break up becomes more frequent after the 4th month and myocytes appear increasingly atrophic, which facilitates the extension of fibrosis. This is accompanied at times by a thinning of the arterial wall with or without localized disappearance of the media. All these modifications are discussed and compared to what we had previously found in the femoral artery [12].

Aging↗

Fine structure of the glomerular basement membrane of the rat kidney visualized by high-resolution scanning electron microscopy.

The fine structure of the glomerular basement membrane (GBM) of the rat kidney was studied by means of high resolution scanning electron microscopy. Specimens were taken from kidneys perfused with paraformaldehyde, freeze-fractured and then processed with conductive staining. The fractured surface of glomerular tufts exhibited the inner and outer surface of the GBM uncovered by endothelial and epithelial cells. The lamina densa was composed of densely packed granular material together with scattered fibrils. The laminae rarae interna and externa were composed of a meshwork that showed some structural heterogeneities. The meshwork composing the lamina rara interna contained 5- to 9-nm-thick fibrils, had pores 11-30 nm wide, and was associated with granular material except in those places that corresponded with endothelial fenestrae. The meshwork of the lamina rara externa was made up of 6- to 11-nm-thick fibrils, and had smaller pores under the foot processes (10-24 nm wide) than those near the filtration slits (16-32 nm wide). In addition to the meshwork, the lamina rara interna contained microfibrils that were arranged differently depending on the topography of the capillary wall: scattered fibrils had no predominant orientation at the convex side, circumferential bundles lay at the concave side of the peripheral capillary wall, and had a circumferential arrangement in the paramesangial wall.

Animals↗

Long-term endoneurial changes after nerve transection.

Long-term endoneurial changes in the distal stump of transected rat sciatic nerve were examined from 8 to 50 weeks after nerve transection. The morphological alterations were followed both in nerves which were allowed to regenerate and in nerves in which regeneration was prevented by suturing. The nerves prevented from regenerating showed markedly atrophied Schwann cell columns after 20 weeks and a disappearance of some Schwann cell columns after 30 weeks. The surrounding endoneurial fibroblast-like cells gradually lost their delicate cytoplasmic extensions and formed rough fascicles around numerous shrunken Schwann cell columns or around areas from which Schwann cells had apparently disappeared. Inside the fascicles, the Schwann cell loss was replaced by collagen fibrils or occasionally, by a dense accumulation of microfibrils. The loss of endoneurial cytoplasmic processes continued up to 50 weeks, leaving behind patches of thin fibrils around numerous shrunken Schwann cell columns or around collagenous areas where Schwann cells were lost. The endoneurial matrix showed presence of thin 25- to 30-nm collagen fibrils close to shrunken Schwann cell columns up to 50 weeks but in areas with advanced degeneration a shift towards regular 50- to 60-nm collagen fibrils occurred. The degenerated areas resembled those described in Renaut bodies and neurofibromas. Despite suturing of transected nerves to prevent sprouting, occasional regenerating sprouts were noted in the Schwann cell columns. These axons were surrounded in a sheath-like fashion by pre-existing endoneurial cell fascicles covered by a basal lamina. In the reinnervating nerves the endoneurial space gradually lost its compartmentized structures consisting of collagen fibrils and endoneurial fibroblast-like cells. After 20 weeks the endoneurial cells were inconspicuous and the extracellular matrix consisted mainly of 50- to 60-nm collagen fibrils. During axonal growth and maturation, Schwann cells containing unmyelinated axons surrounded large, myelinated axons in a collar-like fashion. Close to these collars of Schwann cells, thin 25- to 30-nm collagen fibrils were noted in focal areas, even after 50 weeks. Occasionally, numerous clusters of regenerating axonal sprouts were noted in the perineurium. These were surrounded by multiple layers of cells possessing basal lamina. The present results show that after nerve transection the distal stump of the severed nerve shows dynamic changes in the endoneurial space, especially in nerves where reinnervation is prevented.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Fine structure of psammoma bodies at the outer aspect of blood vessels in meningioma.

Psammoma bodies at the perivascular area in five cases of meningioma were examined with the electron microscope. In general, meningocytic cells invest the outer aspect of blood vessels, which are constituted by multilayered basal laminae, collagen fibers, microfibrils, and pericytes. Remnants of degenerated cells are observed in some areas of the perivascular space. Matrix vesicles and matrix giant bodies with or without mineralized deposits are also present in those areas. Energy dispersive, X-ray microanalysis evidenced the presence of both calcium and phosphorus (probably hydroxyapatite) mineralized precipitates. Production of psammoma bodies in the perivascular area may indicate that matrix vesicles and matrix giant bodies are derived from degenerated cells, which then sequestrate hydroxyapatite crystals to form psammoma bodies.

Blood Vessels↗

Subperineurial space of the sural nerve in various peripheral nerve diseases.

The size of the subperineurial space of the sural nerve has been evaluated quantitatively in 69 cases of various peripheral nerve diseases and in controls. A significant increase was found in beriberi neuropathy (6 cases) and idiopathic polyradiculoneuropathy (9 cases) as compared with the control (8 cases). On electron microscopy a few macrophages, fibroblast processes, collagen fibrils with a diameter of 50 nm, microfibrils with a diameter of 8 nm, and amorphous material were observed in both the enlarged subperineural space and the endoneurial intercellular space. They were less frequently observed in controls. No significant correlation was found between the size of subperineurial space and the density of myelinated fibers.

Adolescent↗

Distribution of collagen types I and III and basal lamina in human gastric carcinoma: an immunohistochemical and electron microscopic study.

Collagen types I and III were examined immunohistochemically in 32 cases of gastric carcinoma classified as poorly differentiated adenocarcinoma with scirrhous stroma, well differentiated adenocarcinoma with intermediate stroma, or poorly differentiated adenocarcinoma with medullary stroma. In the stroma of scirrhous carcinoma, types I and III collagens were distributed abundantly in fibrillar or granular patterns with little difference in the intensity of staining. In well differentiated adenocarcinoma, type I collagen was diffusely distributed in the stroma with type III collagen distributed sparsely. In poorly differentiated adenocarcinoma with medullary stroma, the two types of collagen were only found around capillaries, constituting the tumor interstitium. Electron microscopic examination of scirrhous carcinoma showed tumor cells partially covered with fibroblasts, and discontinuous basal lamina, collagen fibers and microfibrils present between tumor cells and fibroblasts. In well differentiated carcinoma, tumor cells were surrounded by fibroblasts, and well developed basal lamina was observed beneath the tumor cells. In poorly differentiated carcinoma with medullary stroma, the stroma consisted of capillaries and very few fibroblasts with discontinuous basal lamina occasionally being present between tumor cells and fibroblasts.

Adenocarcinoma↗

Ultrastructural visualization of elastic fibres with a tannate--metal salt method.

A modification of the tannic acid-metal salt method was applied as an ultrastructural stain for elastin. Thin sections of glutaraldehyde-fixed, embedded rat aorta and rabbit elastic cartilage, with and without osmication, were examined. Raising the pH of the tannic acid solution from 2.7 to 9.0 progressively increased the electron-density of elastic fibres and collagen fibrils in osmicated and unosmicated specimens. The maximum tannic acid staining of elastic fibres was observed in the pH range 7.0-9.0. Collagen staining, although less intense than that of elastic fibres, was also greatest in this pH range. Elastic fibres in osmicated specimens demonstrated the strongest tannic acid staining with a minimal increase in density of collagen and cell nuclei when compared to the unosmicated specimens. Sequential treatments of osmicated specimens with tannic acid pH 7.0-9.0, and uranyl acetate, pH 4.1, enhanced the density of the elastin intensely, increased collagen staining moderately, but hardly increased the density of nuclei and microfibrils. In elastase-digested osmicated specimens, all tannic acid (pH 7.0)-uranyl acetate-reactive elastin was selectively removed. These results demonstrate that all the neutral and alkaline tannic acid-uranyl acetate methods can be used as a postembedment stain for elastin specimens fixed in glutaraldehyde and osmium tetroxide.

Animals↗