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

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*

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

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

A defective HIV-1 helper virus DNA, pHyPC, was assembled by deleting the RNA packaging signal, env, nef and the 3'LTR sequences. HIV-1 like virus particles that carry the HIV-1 receptor, CD4 were generated by coexpression of pHyPC and plasmid DNAs encoding different chimeric CD4 proteins. The CD4 particles, sharing the CD4 ectodomain, precisely fused to different membrane anchors. CD4(+) particles specifically bound to HIV-1 Env expressing cells, but any signs of infection into these cells were not detected. Binding was only partially blocked by either polyclonal anti-CD4 antibodies or by high concentrations of soluble CD4. Suprisingly, CD4(+) particles also adsorbed to HeLa, CHO, NIH3T3 and COS-7 cells in the absence of HIV-1 Env expression. Adsorption was comparable in strength and speed to the highly specific CD4-Env interaction. CD4(-) particles exhibited only background levels of binding. Cell binding was CD4- dependent, but it was independent of the cell type from which the CD4(+) particles originated. Interestingly, CD4-dependent/Env-independent binding was only found when CD4 was present on virus particles. This suggests that the micro-environment of CD4 on virus particles uniquely expose this new cell binding activity. Its high affinity could explain in part why infection of Env(+) cells by CD4(+) particles was not detected. Further experiments will be required to evlauate whether this strong membrane interaction could represent one step in the multiple-step viral entry process.
Adsorption ; Animals ; Antibodies ; COS Cells ; DNA ; Helper Viruses ; HIV-1* ; Membranes ; Plasmids ; Product Packaging ; RNA ; Virion

Herpes zoster is known to recur only rarely. However reports on the recurrence rates by various authors ranges from 0.l% to 8%. We have experienced 2 cases of recurrent herpes zoster in a 63-year-old female and a 62-year-old male who presented with typical clinical features and previous history of herpes zoster. They were suffering from breast cancer and Hodgkin's disease, respectively. By employing negative and positive staining methods of electron microscopy, typical herpes viral particles were demonstrated from the vesicular fluid.
Breast Neoplasms ; Female ; Herpes Zoster* ; Hodgkin Disease ; Humans ; Male ; Microscopy, Electron* ; Middle Aged ; Recurrence ; 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

To evaluate the ultrastructure of cornea keratocyte infected by Herpes simplex virus, the cornea was excised from enucleated rabbit eyes. Cornea epithelium and endothelium of these was removed and cultivated in culture media. After 7 days, the Kos strain of Herpes simplex virus was inoculated into the cultured cornea keratocytes and co-cultivated until the cytopathic effect occurred. Cornea keratocytes was fixed in the 3%glutaraldehyde and examined with electron microscope. The infected cells were active cells and slightly more round appearance than normal cells. They had microvillies composed of cytoplasmic process protru-sions into collagen fibers. Virus particles was transmitted into neighboring cells easily since there was no barrier function of collagen fibers that was destroyed in the condition of culture. The nuclear changes of infected cells were marked :nuclear chromatin was degraded, condensated and displaced toward the nuclear membrane. There were many virus particles in the nucleus. There were also many degenerations in the cytoplasm, destructed numerous cytoplasmic organelles and many virus particles seen in the cytoplasm. The ultrastructural findings of keratocytes co-cultivated with Herpes simplex virus were identified from our result.
Chromatin ; Collagen ; Cornea ; Culture Media ; Cytoplasm ; Endothelium ; Epithelium ; Herpes Simplex* ; Microvilli ; Nuclear Envelope ; Organelles ; Simplexvirus* ; Virion

From June to October in 1993~1995, cultured Penaeus chinesis and Penaeus japonicus occurred mass mortality at the farm in Western Sea of Korea. The disease was reproduced in healthy shrimp by injection of filtered (at 0.45 micromeger) homogenate of infected shrimp. So we concluded hat it was filterable agents like virus. Clinical symptoms were white spots on the carapace and reddish tail. Histopathological changes were characterized by hypertrophied nuclei at cuticular epidermis lymphoid organ, hematopoietic tissue. In negatively stained preparation, the virion revealed rod-shaped, enveloped, nonoccluded. Cytopathic effect (CPE) were not observed by virus in CHSE-214, RTG-2, EPC, FHM cell lines. Base on the above facts, the reason of mass mortality of penaied shrimp was baculovirus.
Baculoviridae ; Cell Line ; Dental Caries ; Epidermis ; Korea ; Mortality ; Penaeidae* ; Tail ; Virion ; Virus Diseases*

Considerable effort has been directed at understanding the structure and function of HIV-1 envelope glycoproteins. It has been difficult to characterize HIV-1 envelope glycoproteins due to the limited availability of these proteins from virus particles or infected cells. To facilitate the structural and functional analysis of HIV-1 envelope glycoproteins, recombinant baculoviruses were generated to express wild type or mutant HIV-1 envelope glycoproteins. The gp160 precursor protein as well as the gp120 glycoprotein were detected in the cell infected with recombinant BacENVw.t containing wild type HIV-1 envelope gene. In the insect cells infected with recombinant BacENVc with mutations at the cleavage site of gp160, a precursor form of envelope glycoprotein was produced, but not secreted into the culture medium. However, the insect cells infected with recombinant BacENVc/t containing both mutations at the cleavage site and membrane spanning region produced mutant envelope glycoproteins that were efficiently secreted into the culture medium in the form of precursor. Therefore the recombinant HIV-1 envelope glycoproteins produced in this system would be useful as immunogens in the development of a vaccine against AIDS.
Baculoviridae* ; Glycoproteins* ; HIV* ; HIV-1* ; Humans* ; Insects ; Membranes ; Staphylococcal Protein A ; Virion