To identify herpesviral virions secreted by viral replication, we have established the nuclease-resistance assay (NRA) as a comparative analysis to other conventional assays, including electron microscopy (EM). For this study, we used an efficient experimental in vitro infection model for Alcelaphine herpesvirus 1 (AlHV-1), a gamma herpesvirus, to propagate the virus. The NRA could identify extracellular, cell-free, enveloped virions in the supernatants after 24 hours post inoculation (h.p.i.) of AlHV-1. The results of EM observation were correlated with those of NRA. Mature virions were observed in the clarified, concentrated supernatants from 24 h.p.i. by EM. These results show that sensitivity of the NRA is comparable with that of EM for the identification of mature enveloped virions, which directly presents evidence of herpesviral lytic replication. NRA allows us to differentiate the virus from other member of Herpesviridae, and has extended the possibility of analysis for quantification of shedding viruses when used in conjunction with real-time polymerase chain reaction (PCR).
Herpesviridae ; Microscopy, Electron* ; Real-Time Polymerase Chain Reaction ; Virion*

Adenoviruses are icosahedral virions containing double-stranded linen DNA. They are 70 nm to 90 nm in diameter and capsid is composed of 252 capsomeres. Several members of this group, including types commonly associated with respiratory disease in man, are capable of producing malignant tumors in young hamsters and a few types have been shown to be oncogenic in young rat. Previous report involving effect of caffein on transformation induced by Adenovirus type 12 [9] has been carried out. The present report represents a continuation of previous study. To obtain evidence concerning the of(tract of MNNG (N-methyl-N'-nitro-N-nitroguanidine) on transformation, investigation of adenovirus type 12 of this group was undertaken. For practical consideration it was desirable to investigate the effect of MNNG on the adenovirus type 12-induced transformation in L cell. Results were as following 1. Adeno virus type 12 induced transformation was enhanced in the presence of MNNG. 2. Yields of adeno type 12 virus in L cell were slightly inhibited by treatment of MNNG.
Adenoviridae* ; Animals ; Bedding and Linens ; Capsid ; Cricetinae ; DNA ; Methylnitronitrosoguanidine ; Rats ; Virion

Standard plaque assay using agarose-overlay has long been used for titration of many infectious virus particle. Plaque assay for the titration of varicella-zoster virus and its live vaccine requires three intermittent agarose overlay to visualize plaques. Overall procedure of the assay takes at least nine days from virus inoculation and microbe contamination including fungi is frequently accompanied during incubation period. We studied whether an immunofocus assay in conjunction with peroxidase-mediated immunohistochemical reaction may replace the standard plaque assay for the virus titration by comparing the two methods. A linear relationship was observed between number of foci and virus dilution. The number of foci in a given dilution of virus appeared a little higher than counted plaques formed in standard plaque assay. Independent titration results obtained from two assay methods for a given dilution of virus demonstrated a strong correlation (r2=0.99). Foci of virus infected cells as revealed by the enzyme reaction could be counted either 4 days post-infection (p.i.) under low magnification (40X) microscopy, or 6 days p.i. by naked eye observation. Larger size of cell cuture plate, virus adsorption at 35 degrees C, and 10% FBS in diluent appeared to be better conditions for the assay. Immunofocus assay will be an effective and dependable titration method for varicella-zoster virus and its live vaccine in place of the standard plaque assay in respect to accuracy, costs, and experimental convenience.
Adsorption ; Fungi ; Herpesvirus 3, Human* ; Microscopy ; Sepharose ; Virion

We report herein a case of bowenoid papulosis, which developed on the groin, the shaft of the penis and left parietal area, of scalp in a 31-year-old male. Clinical features present as brown to brown-black papules and verrucous plaques ranging from 2 cm to 12 cm in diameter. Electron microscopic examination of the specimen obtained from our patient fail to reveal viral particles.
Adult ; Genitalia* ; Groin ; Humans ; Male ; Penis ; Rabeprazole ; Scalp ; Virion

Epidermal cysts may develop on any part of the body. Most of them are thought to occur following inflammation of the epithelium of the hair follicle. These found on the palm and sole where the hair follicle is absent have been considered to developmant of following a traumatic inclusion of the epidermis into the Dermis. Because only a few cases w re known to be related to preceding trauma, the latter assumption has been questioned. Reerly, the HPV-like virions and papillomavirus genus-speciric antigen were detected in the epicrml cyst of the sole in some reports. 1 our cases of plantar epidermal cyst were studied for the presence of human papillomavirus using conventional histologic and immunohistochemical examinator Histologic examination showed three characteristic findings, that is, intracytoplasmic eosinophil odies in the cyst wall, parakeratosis within the cyst caviti, and vacuolar structures disperse the wall and cavity. In all of the cases, immunohistochernical staining was positive for papule evirus antigen. These findings suggest an etiologic as.iation between the papillomavirus infection and plantar epidermal cyst.
Dermis ; Eosinophils ; Epidermal Cyst* ; Epidermis ; Epithelium ; Hair Follicle ; Humans* ; Inflammation ; Papilloma* ; Papillomavirus Infections ; Parakeratosis ; Virion

This study was performed to investigate the morphological changes of the human conjunctival epithelium when the cultured conjunctival tissue was co-cultivated with the inoculated adenovirus. Specimens swabbed from the lower conjunctival sac of patients with epidemic Keratoconjunctivitis were inoculated into the cultured conjunctival epithelial cells and co-cultivated. The adenovirus-infected cell revealed viral particles, crystalline arrays of adenovirus, paracrystalline inclusions and three types of inclusions within the nucleus. The cytoplasm did not show the specific pathologic changes. These results suggest that adenovirus can be co-cultured with conjunctival epthelial cell and the infected cell showes the typical morphological changes within the nucleus. This in-vitro co-cultivation of adenovirus with conjunctival epithelial cell may be used for an in-vitro model in the research of adenovirus.
Adenoviridae* ; Crystallins ; Cytoplasm ; Epithelial Cells* ; Epithelium ; Humans ; Inclusion Bodies ; Keratoconjunctivitis ; Virion

A 79-year-old man with balanitis circumscripta plasmacellularis(BCP), presenting as an erythematous constricting band of the inner surface of the prepuce encircling the penile shaft is described. The biopsy specimen of the lesion showed, in addition to the typical histologic findings of BCP, increased fibrosis and decreased amount of elastic fibers which correlate well with our clinical observations. Electron microscopic examination revelaed no viral particles or elastic fibers. Immunohistologically, IgG was found to be the major immunoglobulin class in the plasma cellular inf iltrate.
Aged ; Balanitis* ; Biopsy ; Elastic Tissue ; Fibrosis ; Humans ; Immunoglobulin G ; Immunoglobulins ; Male ; Plasma ; Virion

The effective inactivation of microorganisms in drinking water by Ultraviolet (UV) irradiation is regarded as a new low-cost water treatment method shoeing high removal rate of relatively stable infectious virus particles including poliovirus. In the present study, we examined virus inactivation by UV in various water environments. Samples were collected from finished water and surface water, and tested for turbidity. UV dose of 18, 22, 30, 36 and 40 milli-Joule (mJ)/cm2 were used by combination of 2 mW/cm2 UV intensity and time of 9, 11, 15, 18 and 20 second. Depths of water were fixed at 0.37 cm and 8 cm, and virus titers were shown by plaque forming unit (PFU). Poliovirus was inactivated to 99.0% by 18 mJ/cm2 of UV dose in the condition of 0.08 Nephelometry Turbidity Unit (NTU) and 8 cm depth of water. Poliovirus at 30 mJ/cm2 of UV dose under the same condition was inactivated to 99.7%. Furthermore, Poliovirus at 56.60 NTU and 8 cm depth of water was inactivated to 92.0% and 98.5% by 18 mJ/cm2 and 30 mJ/cm2 of UV dose, respectively. The degrees of virus inactivation were dependent upon the UV dose, the turbidit, y and the depth of water. In conclusion, introduction of UV disinfections can be considered in drinking water purification systems in case reasonable engineering support is possible.
Drinking Water ; Nephelometry and Turbidimetry ; Poliovirus ; Shoes ; Virion ; Virus Inactivation ; Water Purification ; Water*

The effective inactivation of microorganisms in drinking water by Ultraviolet (UV) irradiation is regarded as a new low-cost water treatment method shoeing high removal rate of relatively stable infectious virus particles including poliovirus. In the present study, we examined virus inactivation by UV in various water environments. Samples were collected from finished water and surface water, and tested for turbidity. UV dose of 18, 22, 30, 36 and 40 milli-Joule (mJ)/cm2 were used by combination of 2 mW/cm2 UV intensity and time of 9, 11, 15, 18 and 20 second. Depths of water were fixed at 0.37 cm and 8 cm, and virus titers were shown by plaque forming unit (PFU). Poliovirus was inactivated to 99.0% by 18 mJ/cm2 of UV dose in the condition of 0.08 Nephelometry Turbidity Unit (NTU) and 8 cm depth of water. Poliovirus at 30 mJ/cm2 of UV dose under the same condition was inactivated to 99.7%. Furthermore, Poliovirus at 56.60 NTU and 8 cm depth of water was inactivated to 92.0% and 98.5% by 18 mJ/cm2 and 30 mJ/cm2 of UV dose, respectively. The degrees of virus inactivation were dependent upon the UV dose, the turbidit, y and the depth of water. In conclusion, introduction of UV disinfections can be considered in drinking water purification systems in case reasonable engineering support is possible.
Drinking Water ; Nephelometry and Turbidimetry ; Poliovirus ; Shoes ; Virion ; Virus Inactivation ; Water Purification ; Water*

For the purpose of morphologic analysis of human cytomegalovirus (HCMV) antigens reacting with specific monoclonal antibodies, we observed HCMV particles after immunogold staining. HCMV was cultured in human fetal lung fibroblasts to be concentrated by polyethylene glycol 6,000. The HCMV stock was dropped onto Formva-coated grids and was fixed by 2% glutaraldehyde. The grids were reacted with MCMVA57, 93, 135 or with SCMVM1, 6, 14, 49 monoclonal antibodies (MoAbs) follwed by gold (10 nm)-conjugated goat anti-mouse IgG. Then the grids were stained with 2.5% uranyl acetate to be observed under Hitachi 500 or Jeol 1,200 electron microscope. When HCMV was reacted with SCMVM14 and SCMVM49 MoAbs, gold particles were adsorbed to virion envelopes, suggesting that the reactive antigens were envelope proteins. In cases of MCMVA135 and SCMVM6 MoAbs, gold particles were adsorbed to dense bodies as well as to virion envelope. These results, together with the previous results of immunologic and genetic characterization, suggested that the reactive antigens of MCMVA135 and SCMVM6 MoAbs were gB homologue and structural protein, respectively. In case of SCMVM1 MoAb, gold particles were adsorbed to capsids, envelopes, and dense bodies, suggesting that the reactive antigen was structural protein. In case of SCMVM8 MoAb, gold particles were observed between the envelopes and capsids, which space was supposed to be the tegument, suggesting that the reactive antigen was carbohydrate moiety of glycoprotein or its polymer. In cases of MCMVA57 and MCMVA93 MoAbs, gold particles were adsorbed to only dense bodies, suggesting that the reactive antigens were precursors of structural proteins.
Antibodies, Monoclonal* ; Capsid ; Cytomegalovirus* ; Fibroblasts ; Glutaral ; Glycoproteins ; Goats ; Humans* ; Immunoglobulin G ; Lung ; Polyethylene Glycols ; Polymers ; Virion