SUBACUTE SCLEROSING PANENCEPHALITIS WITH SPECIAL REFERENCE TO THE ULTRASTRUCTURE OF INCLUSIONS IN THE BRAIN AND LUNG

A 7-year-old boy, who was diagnosed as typical SSPE by clinical data and laboratory findings, was autopsied and observed by immunofluorescent techniques, light and electron microscope. The morphological characteristics in the brain were perivascular cuffings with plasma cells, lymphocytes and mononuclear cells, gliosis and a large number of intranuclear and intracytoplasmk inclusions in the neuroglias and nerve cells. Various kinds of intranuclear inclusions were elucidated by electron microscopy and the fine structures of these inclusions were described in detail. At least five types of intranuclear inclusions were regarded as specific in SSPE. The presence of intranuclear inclusions of mononuclear cells in the lungs resembling the inclusions in the neuroglias suggested that the disease was not localized in the brain but could be disseminated throughout the body.     

SUBACUTE SCLEROSING PANENCEPHALITIS

An autopsy case of subacute sclerosing panencephalitis (SSPE) in a 5-year-old boy, with rapid progression to a comatose state in 2 weeks after the onset of right hemiplegia, is described. The levels of antibody to measles virus in the serum and the cerebrospinal fluid were increased, and high levels of IgG in the latter were found. A characteristic pattern of electroencephalogram (EEG) showing periodic suppression of high voltage complexs was also found during the course of the disease. Microscopical examination revealed perivascular cuffing, numerous hypertrophied astrocytes with a diffuse gliosis and sporadic intranuclear inclusions in the brain. In addition to these typical findings of SSPE, impaired cellular immunity was recognized by delayed skin test in vivo , and pathologically severe atrophy of thymus, and follicular atrophy of spleen with amyloid deposition in the wall of the sheathed arteries were found.     

Giant Cell Pneumonia: Light Microscopy, Immunohistochemical, and Ultrastructural Study of an Autopsy Case

An autopsy case of measles giant cell pneumonia with intranuclear inclusion bodies is reported. This case of giant cell pneumonia was studied by light microscopy and immunohis-tochemistry using monoclonal and polyclonal antibody to measles and by electron microscopy (EM). Light microscopic examination showed multinucleated epithelial giant cells with intranuclear and intracytoplasmic inclusions. The giant cells contained prominent, sharply marginated, eosinophilic intranuclear inclusions typical of classic measles pneumonia. Presence of measles antigen was confirmed using both monoclonal and polyclonal antibodies by peroxidase antiperoxidase method. Monoclonal antibody stained positively for intracytoplasmic and intranuclear inclusions. Electron microscopic examination of lung tissue showed intranuclear inclusions of filamentous or worm like nucleocapsid materials in multinucleated epithelial giant cells. The results suggest that this is a case of measles giant cell pneumonia and the intranuclear inclusion bodies are measles viral particles.     

Intranuclear inclusions in a case of pigmented villonodular synovitis of the ankle

Pigmented villonodular synovitis (PVNS) is a proliferative disorder of the synovial lining of the joints. Although the cytological findings of this disorder have been described in the literature, there is no mention of intranuclear cytoplasmic inclusions in this entity. A 65-yr-old woman presented with a painful swelling on the ankle. In addition to other characteristic findings of PVNS, we found the presence of intranuclear cytoplasmic inclusions in the fine-needle aspiration (FNA) smears. The characteristic cytological findings of abundant mononuclear cells occurring singly and in papillary clusters, multinucleated giant cells, and hemosiderin deposition should aid in the diagnosis of PVNS and in its differentiation from melanoma and other soft tissue sarcomas, even in the presence of uncommon features such as intranuclear inclusions.     

Three-dimensional structural analysis of papillary thyroid carcinoma nuclei with serial block-face scanning electron microscopy (SBF-SEM)

Nuclear morphology of carcinoma cells is critical for the pathological diagnosis of papillary thyroid carcinoma (PTC). However, three-dimensional architecture of PTC nuclei is still elusive. In this study, we analyzed the three-dimensional ultrastructure of PTC nuclei using serial block-face scanning electron microscopy which takes advantage of the high-throughput acquisition of serial electron microscopic images and three-dimensional reconstruction of subcellular structures. En bloc-stained and resin-embedded specimens were prepared from surgically removed PTCs and normal thyroid tissues. We acquired two-dimensional images from serial block-face scanning electron microscopy and reconstructed three-dimensional nuclear structures. Quantitative comparisons showed that the nuclei of carcinoma cells were larger and more complex than those of normal follicular cells. The three-dimensional reconstruction of carcinoma nuclei divided intranuclear cytoplasmic inclusions into “open intranuclear cytoplasmic inclusions” connecting to cytoplasm outside the nucleus and “closed intranuclear cytoplasmic inclusions” without that connection. Cytoplasm with abundant organelles was observed in open inclusions, but closed inclusions contained fewer organelles with or without degeneration. Granules with a dense core were only observed in closed inclusions. Our observations suggested that open inclusions originate from nuclear invaginations, and disconnection from cytoplasm leads to closed inclusions.     

Fine-needle aspiration cytology of pleomorphic hyalinized angiectatic tumor: A case report

Pleomorphic hyalinized angiectatic tumor (PHAT) of soft parts is a neoplasm characterized by spindle and pleomorphic cells associated with an angiectatic vasculature. We describe the cytological findings of a fine-needle aspiration biopsy (FNAB) from the right medial knee of a 45-yr-old woman. The aspirate material was entirely submitted in Cytolit solution. The specimen was moderately cellular and was comprised of spindle cells in a background of fibrinous material. The cells varied from small, bland spindle cells with a fine chromatin pattern and inconspicuous nucleoli to larger pleomorphic cells with coarser chromatin and occasional intranuclear inclusions. Most of the cells were arranged singly with sporadic small cluster formation with indistinct cell borders. Rare mononuclear inflammatory cells morphologically compatible with mast cells were identified. The differential diagnosis include solitary fibrous tumor (SFT) and ancient schwannoma, which also shows fibrous-like material and spindle cells that may have intranuclear inclusions.     

Early appearance and localization of intranuclear inclusions in the segments of renal proximal tubules of rats following ingestion of lead.

Intranuclear inclusions in epithelial cells lining rat renal proximal tubules were detected by electron microscopy as early as 4 days after the addition of lead (5 mg/ml as lead acetate) to the drinking water. At 9 and 12 weeks pathological changes, but very few intranuclear inclusions, were apparent in the epithelial cells lining the third segment of the proximal tubules in the outer stripe of outer zone of the medulla. On the other hand, morphological changes were less in the epithelial cells lining the first and second segments of the proximal tubules in the cortex, which contained many intranuclear inclusions. These findings suggest that lead incorporated into the epithelial cells in the proximal tubules may exist in an inert chemical form as inclusions, especially in the second segment. These bodies, therefore, may protect the cells from the toxic effect of lead.     

Ultrastructural study of intranuclear inclusion bodies of pulmonary adenocarcinoma

Intranuclear inclusion bodies are sometimes observed in pulmonary adenocarcinoma by light microscopy. Electron microscopic characteristics of lung cancer cells with intranuclear inclusion bodies were studied. In addition, polymerase chain reaction (PCR) was performed using primers coding for human papillomavirus (HPV) types 16, 18, and 33. Eosinophilic intranuclear inclusion bodies were observed in 22 out of 285 cases by light microscopy. Immunohistochemically, cancer cell nuclei stained with PE-10. Three types of intranuclear inclusion bodies were classified electron microscopically. Type A showed aggregation of electron dense particles (30-40 nm) with an electron-dense core and was most frequently observed. Type B consisted of a mass of branching and whirling tubular structures. Type B intranuclear inclusions had a relationship with inner nuclear membrane. In type C, several spherical inclusions were observed in one nucleus. HPV DNA was detected using PCR and type-specific probes in a case with type A inclusion bodies. This study suggests that intranuclear inclusion bodies in pulmonary adenocarcinoma are formed by several different mechanisms.     

Morphological and virological investigations of cell strains cultured from the brain in Jakob-Creutzfeldt disease and subacute sclerosing panencephalitis.

Cell strains were established in culture from fragments of the brain from 2 cases each of Jakob-Creutzfeldt (JC) disease and subacute sclerosing panencephalitis (SSPE). After about 12 weeks strains from the former spontaneously formed persistent heaped up nodules of cells which appeared to produce reticulin-like fibrils as well as confluent sheets of rounded and spindle, fibroblast-like cells. Similar sheets of cells were obtained from the cases of SSPE but the only nodules formed were smaller and ephemeral. Attempts to detect virus in all 4 strains were made by inoculation of supernatant fluids into cultures of other laboratory cells, haemadsorption, co-cultivation, electron microscopy and immunofluorescence, and testing for interferon production. No evidence was found by any of these methods of the persistent presence of virus in the strains. Immunofluorescence revealed a probable anti-glial cell IgM autoantibody in one case of JC disease. Morphologically some cells resembled astrocytes and others fibroblasts. Those from JC disease contained more vacuoles and redundant membranes than did those from the cases of SSPE, features that are particularly striking in brain cells in human and animal cases of the spongiform encephalopathies.     

Intranuclear inclusions of the human vas deferens.

Intranuclear inclusions in epithelial cells of the human vas deferens have not been well characterized or widely recognized. We studied their prevalence in an adult male population by light microscopy and characterized them using the techniques of histochemistry, electron microscopy, and electron microprobe analysis. They occurred in all cases studied, consisted of electron dense globules, contained neutral mucosubstances, and occasionally were single membrane bound. Epithelial intranuclear inclusions routinely occur in the vas deferens of sexually mature males, do not disappear with advancing age, and should not be interpreted as a pathologic finding.     

Intraductal Papilloma of the Male Breast: An Ultrastructural and Immunohistochemical Study

A case of intraductal papilloma of the male breast was studied by electron microscopy and immunohistochemistry. The major components of this lesion were the epithelial and myoepithelial cells. Intermediate cells showing ultrastructural features of both cell types were also observed. Squamous metaplasia was noted in many areas. Numerous intranuclear helioid inclusions were seen in the tumor cells. The features of this lesion are similar to those of papillomas of the female breast.     

Intraductal Papilloma of the Male Breast: An Ultrastructural and Immunohistochemical Study

A case of intraductal papilloma of the male breast was studied by electron microscopy and immunohistochemistry. The major components of this lesion were the epithelial and myoepithelial cells. Intermediate cells showing ultrastructural features of both cell types were also observed. Squamous metaplasia was noted in many areas. Numerous intranuclear helioid inclusions were seen in the tumor cells. The features of this lesion are similar to those of papillomas of the female breast.     

Relationship of Ehrlichia canis-Infected Mononuclear Cells to Blood Vessels of Lungs

The lung tissue of six dogs with ehrlichiosis and two control dogs was examined with the electron microscope. Mononuclear cells containing inclusions (morulae) of Ehrlichia canis were adhered at one or more sites to the luminal surfaces of endothelial cells of arterioles or capillaries by way of interdigitations or areas of adherence, or an endothelial cell-bound mononuclear cell was chained to another parasitized or nonparasitized mononuclear cell in the lumen. The bifurcation of arterioles was the most common site at which mononuclear cells clung to the endothelium. Cell-free bodies (morulae), enclosed by a single membrane, were present in the lumens of arterioles. Such cell-free bodies were swollen and vesiculated as compared with intracytoplasmic inclusions (morulae) in mononuclear cells.     

JC virus intranuclear inclusions associated with PML-NBs: analysis by electron microscopy and structured illumination microscopy

Progressive multifocal leukoencephalopathy is a fatal demyelinating disorder caused by JC virus infection. JC virus was recently found to target promyelocytic leukemia nuclear bodies (PML-NBs), punctuate domains in the nuclei. Thus, the virus progenies cluster in dots as intranuclear inclusions (ie, as dot-shaped inclusions). In the present study, both the viral major and minor capsid proteins were expressed from polycistronic expression vectors with a powerful promoter, and formation into virus-like particles (VLPs) was examined by electron microscopy. When the upstream regulatory sequence including the agnogene (nt 275 to 490) was present, capsid protein expression was suppressed, but numerous VLPs were efficiently formed with restricted accumulation to PML-NBs. VLPs were uniform, and the cells were severely degraded. In contrast, when the 5' terminus of the agnogene (nt 275 to 409; 135 bp) was deleted, capsid protein expression was markedly enhanced, but VLPs were more randomly produced in the nucleus outside of PML-NBs. VLPs were pleomorphic, and cell degradation was minimal. JC virus association with PML-NBs was confirmed in human brain tissues by structured illumination microscopy. PML-NBs were shaped in spherical shells, with viral capsid proteins circumscribing the surface. These findings indicate that PML-NBs are intranuclear locations for pathogenic JC virus proliferation. Either the agnogene or its product likely supports efficient progeny production at PML-NBs, leading to subsequent degeneration of host glial cells.     

BK polyomavirus interstitial nephritis in a renal allograft recipient

Human polyomavirus (PV) interstitial nephritis has recently been recognized as a cause of severe renal allograft dysfunction. It occurs in immunosuppressed patients after reactivation of the latent virus PV type BK (BK virus) in the renal epithelium. BK disease is defined as a morphologically manifest renal infection with cytopathic signs accompanied by varying degrees of interstitial inflammatory cell infiltrates and functional impairment. It is also identified by the presence of cells containing viral inclusion bodies (decoy cells) in the urine. The authors report a case of BK PV interstitial nephritis in a 36-year-old renal allograft recipient. Under light microscopy the chief diagnostic indicator was detection of intranuclear viral inclusions, which were found exclusively in tubular epithelial cells. Cells with viral changes were often enlarged with nuclear atypia and chromatin basophilia. Widespread interstitial plasma cell infiltrates associated with tubulitis were present. Intranuclear paracrystalline arrays of virus particles 35-38 nm in diameter were present as characteristic ultrastructural indicators. Urine samples revealed decoy cells with ground-glass-type intranuclear inclusions positive for BK virus by electron microcopy.     

BK Polyomavirus Interstitial Nephritis in a Renal Allograft Recipient

Human polyomavirus (PV) interstitial nephritis has recently been recognized as a cause of severe renal allograft dysfunction. It occurs in immunosuppressed patients after reactivation of the latent virus PV type BK (BK virus) in the renal epithelium. BK disease is defined as a morphologically manifest renal infection with cytopathic signs accompanied by varying degrees of interstitial inflammatory cell infiltrates and functional impairment. It is also identified by the presence of cells containing viral inclusion bodies (decoy cells) in the urine. The authors report a case of BK PV interstitial nephritis in a 36-year-old renal allograft recipient. Under light microscopy the chief diagnostic indicator was detection of intranuclear viral inclusions, which were found exclusively in tubular epithelial cells. Cells with viral changes were often enlarged with nuclear atypia and chromatin basophilia. Widespread interstitial plasma cell infiltrates associated with tubulitis were present. Intranuclear paracrystalline arrays of virus particles 35-38 nm in diameter were present as characteristic ultrastructural indicators. Urine samples revealed decoy cells with ground-glass-type intranuclear inclusions positive for BK virus by electron microcopy.     

Histology and ultrastructure of kidney tissue from ringed turtle doves that ingested lead

Ringed turtle doves (Streptopelia risoria) ingested 4 x 122 mg lead pellets or lead acetate (75 micrograms Pb/g body weight) and their kidneys were examined by histological and electron microscopic techniques. Doves that received lead treatments had readily discernable lead intranuclear inclusion bodies in cells of the proximal convoluted tubules. At 33,000 X, the inclusions had a characteristic dense central core, and outer fibrillary zone. Necrosis in the proximal tubular area was seen as deterioration of cell cytoplasm and reduction in mitochondria. In doves ingesting lead, histological evaluation of proximal convoluted tubular cells revealed acid-fast pinkish granules in cell nuclei. Inclusions from rapidly as 1 dove that died after receiving the first of two lead doses (75 micrograms Pb/g body weight/day) had relatively small intranuclear inclusions in cells of the proximal convoluted tubules.     

A light and electron microscopical study of intranuclear cytoplasmic invaginations in interstitial cell tumours of dogs

Intranuclear cytoplasmic invaginations, so-called "pseudoinclusions", were seen in all 7 interstitial cell tumours identified among 32 consecutive canine testicular tumours examined. The proportion of cells with intranuclear cytoplasmic invaginations varied from less than 1 per cent to 15 per cent. Similar "pseudoinclusions" were not seen in any other canine testicular tumours examined; this finding might be helpful in differential diagnosis, especially in less differentiated tumours. Histologically, the cytoplasmic invaginations appeared as round, eosinophilic, clearly demarcated intranuclear inclusions and stained with periodic acid-Schiff. Affected nuclei were enlarged. The process of progressive cytoplasmic invaginations into the nucleus and replacement of most of the nuclear volume of the neoplastic interstitial cells with cytoplasmic material was examined by transmission electron microscopy. Two or more cytoplasmic invaginations were sometimes present within a single nucleus. The nuclear membrane could be followed continuously around the cytoplasmic invaginations and nuclear pores were present in the membrane covering invaginations. The invaginations contained smooth and rough endoplasmic reticulum, vesicles, lipid vacuoles, myelin figures and disrupted membranous profiles. Bundles of interwoven cytoplasmic filaments were among the organelles seen in the invaginations in one tumour. Possible reasons for the formation of intranuclear cytoplasmic inclusions are discussed and maintenance of the normal ratio of nuclear surface to nuclear volume is suggested as the mechanism promoting formation of inclusions in interstitial cell tumours.     

Biological and biochemical characterization of a latent subacute sclerosing panencephalitis (SSPE) virus infection in tissue culture

The present investigation describes the biological and biochemical properties of a persistent SSPE virus infection. Persistently infected cells were derived by cocultivation of infected brain cells and uninfected Vero cells, and cultures were maintained by normal subculturing methods. No infectious virus was ever released from these cultures, and all attempts to induce infectious virus release were unsuccessful. Biological assays showed that infected cells contained nucleocapsid and salt-dependent hemagglutinin antigens, whereas the normal hemagglutinin appeared not to be present. Electron microscopic examination demonstrated the presence of both intranuclear and cytoplasmic nucleocapsids together with the release of virus particles (defective?) from the cell membrane. Biochemical analysis demonstrated that approximately 90% of the intracellular genomic RNA was defective or subgenomic although a small quantity of infectious genomes was present. It is proposed that the large quantities of defective genomes in the infected cells are the major factor in the maintenance of this persistent infection.     

Ultrastructure of inclusions in peripheral blood mononuclear cells in sarcoidosis.

Bundle-shaped tubular (BST) inclusions were identified by electron microscopy in 1 to 6 percent of peripheral blood mononuclear cells obtained from 10 to 11 patients with sarcoidosis. The frequency of the inclusions within mononuclear cells did not correlate with the clinical status of the patients. The relationship, if any, of BST inclusions to sarcoidosis and the immunologic defects in sarcoid patients remains to be determined.