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Tracheobronchial epithelial multinucleation, viral inclusion bodies and malignant disease.

Patients with malignant disease are known to have an increased incidence of multinucleation in their tracheobronchial ciliated epithelial cells as compared with controls matched by age, sex and smoking habit. A seasonal relationship of viral inclusion bodies in the cilated epithelium of asymptomatic subjects has also been shown and is not related to age, sex and smoking habit. We have conducted an epidemiologic study to determine the possible relationships between these factors. Smears from 4,150 patients with a wide variety of pathologic conditions were examined for the presence of viral inclusions and multinucleated, ciliated epithelial cells. High degress of multinucleation were observed least frequenctly in the summer both in patients with and without known malignancy. Cytoplasmic inclusion bodies were also seen least frequently in the summer and autumn both in patients with and without know malignancy. In the presence of cancer, multinucleated epithelial cells and inclusion bodies were seen more frequently regardless of the season. When the seasonal incidence of multinucleated cells in 155 smears containing viral inclusion bodies was analyzed, it was found that patients without cancer had the lowest levels of multinucleation in the summer, whereas cancer patients had a depressed incidence of multinucleation in the winter and spring. Respiratory viruses may have a specific effect on the ciliated epithelium of cancer patients.

Bronchi

Light microscopy of selected viral diseases (morphology of viral inclusion bodies).

A significant number of viruses produce intracellular changes, inclusion bodies, which do have specific and consistent features important to their identification. The inclusions must fulfill the designated criteria for the specific virus or a member of that group. The specific features of viral inclusions are consistent fixative-dependent artifacts: uniformity and familiarity of staining quality are important to the consistency of recognition and identification. Bouin's or Zenker's-acetic acid are the fixatives of choice, and a well-done hematoxylin and eosin stain offers the greatest uniformity of quality. Viral inclusions are not static objects in a cell and recognition of the range of development is often more important than the appearance of a single inclusion. Generally, most viruses within a group cause identical intracellular reactions and produce similar inclusion bodies, thus permitting the pathologist an accurate prediction of the etiologic agent in clinically undiagnosed or unsuspected viral disease. By correlating viral inclusion identification, clinical history, and physical findings the anatomic pathologist can make the diagnosis of many viral diseases rapidly, inexpensively, and with a high degree of accuracy when compared to the complex, expensive, and time consuming procedures of animal inoculation, tissue culture, or serology.

Adenovirus Infections, Human

Ultrastructural and histological studies of brains of ferrets inoculated with subacute sclerosing panencephalitis: similarities to human disease.

Similarities to human subacute sclerosing panencephalitis (SSPE) were revealed in a study of the brains of ferrets inoculated with a cell associated measles virus originally isolated from an SSPE patient. The similarities were greatest in animals that showed neurological signs 3-4 months after inoculation and had high titers of neutralizing antibodies against measles virus. These included dense core particles, nuclear bodies, alterations of basement membranes of small blood vessels, plasma cells, distorted myelin, and rod-like structures in some nuclei. Other abnormalities seen were Hirano bodies, tubular aggregates in cisternae of endoplasmic reticulum and clusters of cytoplasmic tubules. No cells containing viral inclusion bodies were observed by electron microscopy but cell cultures of the brains of these animals always yielded abundant measles virus nucleocapsids in typical SSPE multinucleated syncytia. These findings suggest that the ferret is a suitable animal model for the study of the pathogenesis of SSPE.

Animals

Hyperthermia and host-cell reactivation of adenovirus 12.

Exposure of cultured human fibroblasts to hyperthermia delayed the host-cell reactivation of UV-irradiated human adenovirus type 12 (AD12). The experimental design consisted of irradiating human AD12 with UV doses ranging from 180 to 1800 ergs/mm2, infecting human cell populations at 37 degrees C, exposing the infected cells for 7 h to 39.5 degrees C and 41.8 degrees C, returning them to 37 degrees C and estimating the frequency of cells with intranuclear viral inclusion bodies (IB) 41 and 89 h after hyperthermia treatment. Hyperthermia reduced the fractions of fibroblasts with viral IB in the 41 h samples. By 89 h the capacity to form IB in the treated cells was comparable to that in control cells. Hyperthermia of 39.5 and 41.8 degrees C for 7 h did not affect the replication of nonirradiated AD12. The pattern of host-cell reactivation of AD12 following hyperthermia was compared to that in DNA repair deficient xeroderma pigmentosum cell populations.

Adenoviruses, Human

Canine distemper virus-associated encephalitis: modification by passive antibody administration.

Gnotobiotic puppies were intraperitoneally infected with R252-canine distemper virus (CDV), and treated with normal serum (NS) or CDV-specific antibody. Resultant central nervous system (CNS) lesions were evaluated by light and immunofluorescence microscopy. Lesions in NS-treated dogs, if present, consisted of small foci of neuronal degeneration and accompanying microgliosis, restricted to grey matter of the frontal lobe. Dogs receiving anti-CDV antibody, in contrast, had extensive lesions in the brain. Laminar cortical necrosis, gliosis and astrocytic activation of the grey matter of the frontal lobe was the most consistent lesion observed. In addition, foci of cerebellar granule cell layer degeneration and necrosis was seen along with segmental poliomalacia of the spinal cord. In several dogs, eosinophilic intracytoplasmic and intranuclear viral inclusion bodies were observe in neurons. The lesions observed in these dogs are compared to previously reported lesions of CDV infection in dogs.

Animals

Replication of IPN virus: a cytochemical and biochemical study in SWT cells.

Although IPN virus failed to multiply at 30 degrees, it replicated at 16 degrees and 22 degrees in SWT cells. At 22 degrees the viral eclipse period lasted nearly 6 hr with maximal virion titers attained by 24 hr, whereas replication at 16 degrees was much slower. The replication of the virion was inhibited by 0.05 mug/ml of AD which did not interfere with the production of reovirus. Biochemical studies revealed that cellular DNA synthesis was markedly reduced (greater than 50%) soon after infection whereas total RNA synthesis was enhanced. The period of rapid increase in RNA synthesis paralleled the exponential production of infectious virus. Viral inclusion bodies, revealed by acridine orange-staining of virus-infected cells (SWT and RGG-2) late in the infectious cycle, were found to contain single-stranded RNA on the basis of their staining characteristics and sensitivity to RNase.

Acridines

Human cell cultures infected by tumor-inducing and non-tumor-inducing viruses, an electron microscopic study.

This study is dealing with the infection patterns of adenovirus type 2 and type 12 on human fibroblast cell lines, KB, WI and MAF. With the exception of Ad -12 leads to WI, many intranuclear viral particles were present. None of these second passages (Ad-2 WI-WI, Ad-12 WI-WI, Ad-2 MAF-MAF, Ad-12 MAF-MAF) was found to have viral production. This indicates that the injections on both WI and MAF cells caused by both Ad-2 and Ad-12 cannot be serially transmitted. However, when these infected cells were used to expose KB cells, significant viral yields were obtained. This shows that the infected cells might still carry the viral specific antigens although no visible virions were observed.

Adenoviridae

Relationship between A-type and C-type particles in cells infected by Rous sarcoma virus.

Chicken cells infected with avian RNA tumor virus often contain small cytoplasmic A-type particles which commonly exist as clusters of 50--100 particles when viewed in thin sections. These particles were found more consistently in Rous sarcoma virus-infected than Rous-associated virus-infected cultures, but were generally present in only a small fraction of the total infected cells. The results of the survey of cells infected with various strains of leukosis-sarcoma viruses led to the hypothesis that the A particles develop in cells undergoing cytopathological degeneration. The hypothesis explains also the evanescent nature of the appearance of these particles in infected cells. The application of immunoelectron microscopic methods using monospecific antisera against viral internal proteins revealed that the A particles contain components immunologically related to the proteins of C-type virus.

Animals

Replication of parainfluenza type 3 virus in alveolar macrophages: evidence of in vivo infection and of in vitro temperature sensitivity in virus maturation.

Approximately 2% of cultured alveolar macrophages (AM), originally lavaged from the lungs of parainfluenza type 3 virus (PI-3V)-infected calves, were observed to contain viral antigen (by fluorescent antibody method) or viral nucleocapsids (by electron microscopy). Plaque assays, however, indicated that virus titers were generally low when cultures were incubated at 37 degrees C for 10 days. AM, obtained from "in vivo infected" and "noninfected" calves, were found to be equally susceptible to further in vitro PI-3V infection when cultures were incubated at 37 degrees C. AM that were obtained from the lungs of normal calves, cultured at 37 degrees C, and inoculated with PI-3V were observed to produce relatively high virus titers when the incubation temperature was shifted down to 32 degrees C. Results from hemagglutinin assays showed that considerable amounts of hemagglutinin were detected when AM cultures were incubated at 32 degrees C, but only limited amounts were detected at 37 degrees C. Results from electron microscopic examinations at both temperatures substantiated the results of plaque and hemagglutinin assays. The PI-3V, isolated from AM cultures incubated at 32 degrees C, grew well in Madin-Darby bovine kidney cells at 32 degrees C, but little virus was produced at 37 degrees C. In contrast, parent PI-3V grew equally well at both temperatures. The results are discussed in terms of host susceptibility, temperature-sensitivity and virus maturation, and surface viral antigens and persistent viral infection.

Animals

Cytomegalovirus: nucleocapsid assembly and core structure.

New ultrastructual aspects of cytomegalovirus nucleocapsid assembly are illustrated in cells from a Kaposi sarcoma. The occurrence of viral deoxyribonucleoproteins in the form of strands and penetration of these strands into open capsids is illustrated. The geometrical arrangement of the core material of the nucleocapsid is aslo emphasized.

Capsid

New perspectives on biliary atresia.

An investigation into the aetiology, diagnosis, and treatment of biliary atresia was carried out because the prognosis remains so poor.In an electron microscopical study no viral particles or viral inclusion bodies were seen, nor were any specific ultrastructural features observed. An animal experiment suggested that obstruction within the biliary tract of newborn rabbits could be produced by maternal intravenous injection of the bile acid lithocholic acid.A simple and atraumatic method of diagnosis was developed using(99) (m)Tc-labelled compounds which are excreted into bile. Two compounds, (99m)Tc-pyridoxylidene glutamate ((99m)Tc-PG) and (99m)Tc-dihydrothioctic acid ((99m)Tc-DHT) were first assessed in normal piglets and piglets with complete biliary obstruction. Intestinal imaging correlated with biliary tract patency, and the same correlation was found in jaundiced human adults, in whom the (99m)Tc-PG scan correctly determined biliary patency in 21 out of 24 cases. The (99m)Tc-PG scan compared well with liver biopsy and (131)I-Rose Bengal in the diagnosis of 11 infants with prolonged jaundice.A model of extrahepatic biliary atresia was developed in the newborn piglet so that different methods of bile drainage could be assessed. Priorities in biliary atresia lie in a better understanding of the aetiology and early diagnosis rather than in devising new bile drainage procedures.

Adult

Isolation and properties of reovirus from cattle in an outbreak of acute respiratory disease.

A cytopathogenic virus was isolated in the primary culture of bovine kidney cells from a nasal swab of affected calves in an outbreak of acute respiratory disease in Japan in 1971. It agglutinated human type O erythrocytes and produced cytoplasmic inclusion bodies. Viral replication was inhibited by 5-iodo-2'-deoxyuridine, indicating that the viral nucleic acid was RNA. The virus was resistant to ether, chloroform, sodium deoxycholate, and acid, and passed readily through Sartorius' membrane filter 100 nm in pore size, but not through the filter 50 nm in pore size. Electron microscopy showed many spherical particles 60 approximately 75 nm in diameter with a double-layered capsid in a sample taken at a buoyant density of 1.34 produced by CaCl equilibrium centrifugation. The virus suspended in 1M MgCl2 solution was stable against heating at 50 degrees C for 30 minutes, but not against freezing at -20 degrees C for 60 minutes. The virus was resistant to, and increased in infectivity after, treatment with 0.063 approximately 1.0% trypsin. These properties were consistent with those established for the reoviruses. Most affected cattle showed a significant rise of antibody titer against reovirus and bovine respiratory syncytial virus, whereas only a few of them presented a serological evidence for recent infection with parainfluenza virus type 3, bovine adenovirus type 7, and bovine parovirus.

Acute Disease