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Syncytium formation by recombinant vaccinia viruses carrying bovine parainfluenza 3 virus envelope protein genes.

The highly syncytium-inducing M strain and the weakly syncytium-inducing SC strain of bovine parainfluenza 3 virus differ by a single amino acid substitution in each of the hemagglutinin-neuraminidase (HN) and membrane (M) proteins, while their fusion (F) proteins are identical (T. Shioda, S. Wakao, S. Suzu, and H. Shibuta, Virology 162:388-396, 1988). We constructed recombinant vaccinia viruses which express separately the M virus HN (Vac-MHN), SC virus HN (Vac-SCHN), M virus M (Vac-MM), SC virus M (Vac-SCM), and common F (Vac-F) proteins. CV-1 cells were infected with the recombinants, singly or in combination, and implanted onto indicator MDBK cells for syncytium formation. Combinations of Vac-MHN plus Vac-F and Vac-SCHN plus Vac-F induced extensive and weak syncytium formation, respectively. Vac-F alone did not induce syncytium formation, and both Vac-MM and Vac-SCM had no effect on syncytium formation. These findings indicated that the syncytium formation by bovine parainfluenza 3 virus requires both the F and HN proteins and that the extensive syncytium formation by the M virus is due to the M virus HN protein. MSC, another weakly syncytium-inducing virus variant, newly isolated from the M virus, was identical to the M virus in the primary structure of the HN and M proteins but differed from the M virus by a single amino acid residue in the F protein. The combination of the recombinant vaccinia virus expressing the MSC virus F protein and Vac-MHN resulted in weak syncytium formation.

Amino Acids↗

Etiology of mumps-like illnesses in children and adolescents vaccinated for measles, mumps, and rubella.

The possible viral etiology of mumps-like illnesses in patients vaccinated for measles, mumps, and rubella (MMR) was studied by use of serum samples prospectively collected, during 1983-1998, from 601 acutely ill Finnish children and adolescents with mumps-like symptoms. Mumps virus was excluded by testing serum samples for mumps antibodies, and the serum samples were further tested for antibodies to adenovirus, enterovirus, Epstein-Barr virus, parainfluenza virus types 1-3, and parvovirus B19. The serum samples of 114 children <4 years old were also tested for antibodies to human herpesvirus 6 (HHV-6). A viral etiology was verified in 84 cases (14%), most commonly Epstein-Barr virus (7%), followed by parainfluenza virus types 1, 2, or 3 (4%) and adenovirus (3%). HHV-6 infection was found in 5 children <4 years old (4%). This study confirms that mumps-like symptoms in MMR-vaccinated children and adolescents are often not caused by mumps virus infection. Careful laboratory-based diagnostic testing of MMR-vaccinated children and adolescents who develop clinical symptoms compatible with those of mumps is important in the treatment of individual patients, in the comprehension of the true epidemiology of these illnesses, and in the evaluation of the impact of MMR vaccination programs.

Adenoviridae Infections↗

Neonatal syncytial giant cell hepatitis with paramyxoviral-like inclusions.

Syncytial giant cell hepatitis in the neonatal period has been associated with many different etiologic agents and may present initially as cholestasis. Infectious causes are most common and include: (1 ) generalized bacterial sepsis, (2) viral agents, (3) toxoplasmosis, (4) syphilis, (5) listeriosis, and (6) tuberculosis. Viral hepatitis may be due to cytomegalovirus, rubella virus, herpes simplex, HHV-6, varicella, coxsackievirus, echovirus, reovirus 3, parvovirus B19, HIV, enteroviruses, paramyxovirus, and hepatitis A, B, or C (rare). Giant cell hepatitis may result in fulminant liver failure with massive hepatocyte necrosis and severe liver dysfunction leading to death, resolution with severely compromised liver function, or liver transplantation. The authors report a 6-week-old male who had an unremarkable perinatal period, became jaundiced after developing diarrhea, and subsequently developed liver dysfunction with massively increased liver enzymes and a coagulopathy. Open wedge and core liver biopsies were performed to determine if the patient should be listed for liver transplantation. Giant cell hepatitis with a significant mixed lymphocytic and neutrophilic infiltrate was present on both the wedge and core biopsies. The residual 60% of hepatocytes had ballooning degeneration and many possessed pyknotic nuclei. The hepatocytes were arranged in a pseudoacinar pattern. Electron microscopy showed paramyxoviral-like inclusions in the giant cells, characterized as large inclusions with fine filamentous, beaded substructures (18-20 nm). Paramyxoviridae are nonsegmented, negative-sense, single-stranded RNA viruses. This family is divided into the Paramyxovirinae subfamily containing respirovirus (Sendai virus, parainfluenza virus type 3), rubulavirus (mumps, parainfluenza virus type 2), and morbillivirus genera (measles); and Pneumovirinae subfamily (pneumovirus genus [respiratory syncytial virus]). Supportive care to determine if hepatic function resolves following the viral episode, liver transplantation with fulminant liver failure, and ongoing evaluation in those who recover to assess chronic liver disease are necessary. Ultrastructural evaluation may unmask the etiologic agent for hepatitis and direct therapy.

Biopsy↗

Influenza and other respiratory virus-related emergency department visits among young children.

BACKGROUND: Influenza and other winter respiratory viruses cause substantial morbidity among children. Previous estimates of the burden of illness of these viruses have neglected to include the emergency department, where a large number of patients seek acute care for respiratory illnesses. This study provides city- and statewide population estimates of the burden of illness attributable to respiratory viruses for children receiving emergency department-based care for respiratory infections during the winter months. METHODS: The number of patients < or = 7 years of age presenting to the emergency department of an urban tertiary care pediatric hospital with acute respiratory infections was estimated by using a classifier based on presenting complaints. The rates of specific viral infections in this population were estimated by using the rates of positivity for respiratory syncytial virus, influenza virus, parainfluenza virus, adenovirus, and enterovirus. Local emergency department market share and US Census data enabled determination of the rates of emergency department visits in the Boston, Massachusetts, area and in Massachusetts. RESULTS: During the 11-year study period, the mean yearly number of patients < or = 7 years of age presenting to the study emergency department during the winter season was 17397. On the basis of the respiratory classifier, the mean number of patients with an acute respiratory infection was 6923, or 398 per 1000 emergency department visits. In the city population, the mean number of emergency department visits for acute respiratory infections was 17906, which is equivalent to 113.9 per 1000 children residing in the city, and in the state population the mean number was 61529, or 94.5 per 1000 children residing in the state. At the state level, 23114 of the visits were for respiratory syncytial virus, 5650 for influenza, 1751 for parainfluenza virus, 2848 for adenovirus, and 798 for enterovirus. For patients 6 to 23 months of age in the state population, there were 19860 emergency department visits for acute respiratory infections, or 168 per 1000 children in this age group, with 6235 visits resulting from respiratory syncytial virus and 2112 resulting from influenza. CONCLUSION: There is a high incidence of emergency department visits for infectious respiratory illnesses among children. This important component of health care use should be included in estimates of the burden of illness attributable to influenza and other winter respiratory viruses.

Boston↗

Epidemiology of viral infections and evaluation of the potential benefit of OM-85 BV on the virologic status of children attending day-care centers.

Viral investigations were performed during 4 winter seasons (88/89, 89/90, 92/93, 93/94) in children attending day-care centers (DCCs) in the Rhône Département in eastern France. Over the total observation period of 4 winter seasons, 780 children were screened with a nasal swab for the presence of viruses. Of those, 230 (29.5%) had a positive viral culture. The viruses identified were respiratory syncytial virus (RSV), influenza A and B virus, parainfluenza virus, coronavirus, rhinovirus, adenovirus and enterovirus. During that time, 83 epidemic events in 47 DCC were recorded. A particular virus was judged to be causally related to an epidemic if the identical virus was isolated in > or = 3 children during the same outbreak of respiratory diseases. Thus, in 51 cases (61.4%) of all epidemics, the following viruses were responsible for an epidemic: RSV (n = 23), coronavirus (n = 10) (only during the season of 1993-1994), influenza A virus (n = 6), rhinovirus (n = 4), enterovirus (n = 4), adenovirus (n = 3) and parainfluenza virus (n = 1). Except for the somewhat surprising accumulation of coronavirus epidemics during the winter of 1993-1994, there were only minor seasonal variations from one year to another. As expected, RSV accounted for about one third of all respiratory tract infections in children attending DCCs and was therefore the most important single causative agent. These results are compared with data from children who did not attend a DCC and were cared for in a private practice.(ABSTRACT TRUNCATED AT 250 WORDS)

Adjuvants, Immunologic↗

Evaluation of cell line 293 for virus isolation in routine viral diagnosis.

Cell line 293, a continuous line of transformed human embryonic kidney cells, has been recognized for its sensitivity in the isolation of adenoviruses, particularly the fastidious species 40 and 41, from stool specimens. To explore the possibility of using this cell line for the isolation of other viruses from clinical specimens, 293 cells were tested for their susceptibility to a variety of viruses including herpes simplex virus, parainfluenza viruses, respiratory syncytial virus, and the enteroviruses ECHO 11, coxsackie B5, and coxsackie B6. All of the viruses induced a cytopathic effect in 293 cells. Consequently, 293 cells were introduced into the diagnostic laboratory and used along with primary African green monkey kidney (AGMK) cell cultures for the inoculation of all respiratory and stool specimens. The study represents a retrospective analysis of the performance of 293 cells over a 22-month period. It was confirmed that 293 cells were more sensitive than AGMK cells for the isolation of adenoviruses from both respiratory and stool specimens. The 293 cells were also sensitive for the isolation of enteroviruses (untyped) but more so from stool specimens than from respiratory specimens. Parainfluenza virus and respiratory syncytial virus were only rarely isolated in 293 cells. Herpesvirus isolates were obtained with equal frequency in both 293 and AGMK cells. This retrospective analysis confirms the value of 293 cells for the isolation of adenoviruses and demonstrates that 293 cells are also useful for the isolation of certain enteroviruses from both respiratory and stool specimens.

Adenoviruses, Human↗

Use of monoclonal antibodies for rapid diagnosis of respiratory viruses in a community hospital.

An indirect fluorescence antibody (IFA) procedure was used for the rapid detection of respiratory viruses in direct clinical specimens and for determining the epidemiology of viruses in a community hospital setting. Viral respiratory diseases were monitored for 10 consecutive respiratory seasons. The Bartels Viral Respiratory Screening and Identification Kit is an IFA method that contains pooled and individual monoclonal antibodies for seven common respiratory viruses. Compared with 8,670 conventional tube cell cultures, IFA staining of direct patient specimens had an overall sensitivity of 84.2% and a specificity of 87.7%. Yearly epidemics of respiratory syncytial virus were seen with alternating short and long intervals between successive periods when virus was isolated. Epidemics following short intervals were more severe. Influenza A virus epidemics occurred yearly, and influenza B virus activity was seen generally every other year. When influenza A and influenza B viruses were cocirculating in a given season, the months of peak activity of one virus were always within 1 month of the peak activity of the other virus. Parainfluenza virus type 1 was detected in the autumn of odd-numbered years, and parainfluenza type 2 virus was seen usually in the autumn of even-numbered years. Parainfluenza type 3 virus and adenovirus were the most ubiquitous agents, with peak incidence occurring in the late winter to spring.

Adolescent↗

Serum antibodies against respiratory tract viruses in episodes of acute otitis media.

Although the findings of epidemiological studies have suggested viral respiratory tract infection (RTI) to be crucially involved in the development of acute otitis media (AOM), the relationship between AOM and viral RTI remains unclear. Serum samples, obtained in the acute and convalescent phases of 57 AOM episodes (in 35 children during the first three years of life) were analysed for IgG antibodies against influenza A viruses, influenza B viruses, parainfluenza virus type 1, respiratory syncytial virus and adenoviruses. One third of the AOM episodes (18/57) could be related to viral RTI, as evidenced by significant increases in viral serum antibody activity. Treatment failure occurred in four AOM episodes where increases in serum viral antibody activity were noted. In three of these failures, antibiotic treatment was unsuccessful despite the bacterial strains not being resistant to the drug used. This suggests that concomitant viral infection may be a determinant of treatment outcome in some AOM episodes.

Acute Disease↗

Methyl prednisolone acetate modulation of infection and subsequent pulmonary pathology in hamsters exposed to parainfluenza-1 virus (Sendai).

Parainfluenza-1 virus (PI-1) has a wide host range; the disease process observed varies with the age, previous exposure, and species. This study was performed to determine possible effects of the corticosteroid methyl prednisolone acetate (MPA) on PI-1 infection in hamsters. Hamsters serologically negative for PI-1 were exposed to virus alone or were exposed to virus the day after pretreatment with a single subcutaneous injection of MPA. Serum antibodies to PI-1 were present in virus-only exposed hamsters by Day 8 and increased up to Day 20. PI-1 was recovered from lungs of virus-only exposed hamsters on Day 2 to 8. Virus antigen was detected by immunocytochemistry on Days 2 to 10 in lungs of virus-only exposed hamsters. Virus-associated lesions in these hamsters began as acute bronchiolar epithelium degeneration and necrosis on Day 4 and were foci of fibrosis by Days 12 to 20. Hamsters exposed to virus after MPA treatment developed no antibodies to virus, had no virus detectable by plaque assays or immunocytochemistry, and had no pulmonary lesions. Hamsters treated with MPA had decreased total lymphocyte counts up to Day 20 after treatment. Treatment of hamsters with MPA one day prior to PI-1 virus exposure is associated with no detectable evidence of viral infection. Humoral and cellular immunity mediated by MPA-sensitive lymphocytes may mediate some of the manifestations of PI-1 pulmonary disease.

Animals↗

Susceptibility of human retinal pigment epithelial cells to different viruses.

BACKGROUND: Different viruses have been reported to be involved in retinal diseases in animal systems. In humans, herpes simplex virus and cytomegalovirus have been found to cause retinal disease. Most of the studied viruses are neurotropic. In this study, the in vitro susceptibility of human retinal pigment epithelial cells (RPEC) to representative members of different groups of human pathogenic viruses was investigated. METHODS: Early cultures of RPE C - after two or three passages - were infected with the following viruses: herpes simplex virus (HSV) type 1, human herpesvirus 6 (HHV-6), Epstein-Barr virus (EBV), cytomegalovirus (CMV), adenovirus types 1 and 7, measles virus, parainfluenza virus and coxsackie virus B3. RESULTS: Cultures of RPE C could be infected with neurotropic viruses like HSV or measles virus as well as with typical respiratory viruses like parainfluenza or adenoviruses. Coxsackievirus, an enterovirus, replicated as well as human CMV, whereas EBV and HHV-6, two lymphotropic viruses, failed to infect RPE. CONCLUSION: These findings suggest that a variety of viruses, including those causing rather common illnesses, might be capable of inducing retinal lesions under certain circumstances due to haematogenous spread during the course of viraemia.

Antibodies, Monoclonal↗

Detection of influenza viruses through selective adsorption and detection of the M-protein antigen.

A model system has been developed which permits rapid detection of influenza viruses through targeting of the M (membrane or matrix)-protein; a type-specific antigen, in an enzyme-linked immunosorbent assay system. This technique exploits the hydrophobic properties of M-protein; the M-protein is selectively and rapidly adsorbed to polystyrene surfaces even in the presence of a 5000-fold excess of bovine serum albumin. Hyperimmune antiserum prepared to purified M-protein is used as the detecting reagent. All type A influenza viruses could be detected by this technique, type B influenza viruses reacted to a slight extent and Sendai virus (parainfluenza virus, type 1) did not react. Virus could be detected to levels as low as 3 ng. Purification of M-protein and preparation of hyperimmune sera from other related virus groups, such as type B influenza viruses, paramyxoviruses and rhabdoviruses should permit detection of these agents by a similar technique.

Adsorption↗

New neplanocin analogues. 6. Synthesis and potent antiviral activity of 6'-homoneplanocin A1.

The design, synthesis, and antiviral activities of 6'-homoneplanocin A (HNPA, 3) and its congeners having nucleobases other than adenine, such as 3-deazaadenine (4), guanine (5), thymine (6), and cytosine (7), were described. Starting from the known cyclopentenone derivative 8, the optically active (mesyloxy)cyclopentene derivative 15 was prepared, which was condensed with nucleobases then deprotected to give target compounds 3-7. Of these compounds, HNPA showed an antiviral activity spectrum that was comparable to, and an antiviral specificity that was higher than, that of neplanocin A. HNPA proved particularly active against human cytomegalovirus, vaccinia virus, parainfluenza virus, vesicular stomatitis virus, and arenaviruses, which is compatible with an antiviral action targeted at S-adenosylhomocysteine hydrolase. HNPA appears to be a promising candidate drug for the treatment of these viruses.

Adenosine↗

New neplanocin analogues. 7. Synthesis and antiviral activity of 2-halo derivatives of neplanocin A.

The syntheses and the antiviral activities of 2-halo derivatives of neplanocin A (1b,c), (6'R)-6'-C-methylneplanocin A (2b), and dehydroxymethylneplanocin A (3b,c) are described. SN2 reaction of the known cyclopentenyl units 12 and 13 with 2-haloadenines under basic conditions gave the protected carbocyclic nucleosides 14b,c and 15b,c, respectively. Starting from the cyclopentenone derivative 5, the optically active tosyloxycyclopentene derivative 11 was prepared, which was similarly condensed with 2-fluoroadenine to give the protected (6'R)-6'-C-methyl derivative 16b. Deprotection of these compounds afforded the target 2-halo derivatives of neplanocin A. Of these new compounds, 2-fluoroneplanocin A (1b) showed an antiviral potency and a spectrum that was comparable to that of neplanocin A (1a). It was particularly active against vaccinia virus, vesicular stomatitis virus, parainfluenza virus, reovirus, arenaviruses (Junin, Tacaribe), and human cytomegalovirus, i.e., those viruses that fall within the purview of the S-adenosyl-L-homocysteine hydrolase inhibitors.

Adenosine↗

Field trial of a quadrivalent vaccine against calf respiratory disease.

A quadrivalent vaccine containing the killed antigens of respiratory syncytial virus, parainfluenza virus type 3, Mycoplasma dispar and M bovis, emulsified with an oil adjuvant, was tested for efficacy against naturally occurring calf respiratory disease. Three batches of beef cattle aged 12, seven and three weeks at the time of first vaccination were used. Within each batch of approximately 100 animals, half were vaccinated subcutaneously on three occasions, three weeks apart and half served as unvaccinated controls. Over the trial period, from November 1981 to May 1982, 27 per cent of the control calves were treated for respiratory disease compared with 16.3 per cent of the vaccinated animals. This reduction of non-fatal disease in the vaccinated animals represented a protection rate of almost 40 per cent and was statistically significant (P less than 0.05). Mortality was also reduced from 3.4 per cent in the control calves to 1.9 per cent in the vaccinated animals but this difference was not statistically significant. During a major outbreak of disease associated with respiratory syncytial virus, the protection rate increased to 69 per cent (P less than 0.01). Furthermore, in the batch of cattle aged seven weeks at first vaccination there was significantly less pneumonic consolidation at death in the vaccinated animals than in the control animals (P less than 0.05).

Animals↗

Injury of respiratory epithelium.

The large surface area provided by the respiratory tract epithelium of humans for exposure to microbial agents and toxic substances in the environment makes this organ system very vulnerable but a good early indicator of adverse health effects. However, the complexity of pulmonary defense mechanisms complicates definition of the interactive effects of pollutants and infectious agents. Tracheal organ culture has been utilized to maintain organized, differentiated respiratory epithelium in vitro. This model system permits the exposure of respiratory epithelium to injurious agents in an easily visualized and controlled environment. Effects of individual toxin and/or infectious agents may be examined without the involvement of most host defenses and unwanted secondary microbial invaders which hamper interpretation of in vivo model studies. Further, elements of host immune response, pharmacologic agents and the like may be added selectively if desired. A body of information is being developed on specialized respiratory cell injury by various common pathogenic agents--including respiratory syncytial virus, parainfluenza virus type 3, Bordetella pertussis and Mycoplasma pneumoniae--through studies in tracheal organ cultures. These agents injure specialized epithelial cells in different ways, providing a spectrum of changes against which the added effects of toxic substances could be evaluated at the cellular and subcellular levels. Information on the pathogenesis of infectious/toxic injury could suggest new directions for human health research and for means to benefit the human host.

Animals↗

Viral etiology and epidemiology of acute lower respiratory tract infections in Korean children.

Viral etiologic agents of acute lower respiratory tract infections were studied from November, 1990, through April, 1994, in Korean children. From 712 children who visited or were admitted to Seoul National University Children's Hospital because of acute lower respiratory tract infections, 804 nasal aspirates were collected; viral agents were detected by virus isolation and virus antigen was detected by indirect immunofluorescent staining. One or more viral agents were identified in 369 (45.9%) cases; of which 3.3% were mixed infections. The pathogens identified were respiratory syncytial virus (27.2%), parainfluenza virus type 3 (7.8%), influenza A virus (3.9%), adenovirus (3.9%), parainfluenza virus type 1 (1.7%), influenza B virus (1.4%), parainfluenza virus type 2 (0.5%), measles virus (0.1%) and others (0.9%). The clinical patterns of viral lower respiratory tract included pneumonia (56.6%), bronchiolitis (35.2%), croup (6.5%) and tracheo-bronchitis (1.6%). Infections with respiratory syncytial virus, parainfluenza virus types 1 and 3 and influenza A and B virus occurred in epidemics, whereas adenovirus was isolated sporadically throughout the study period. The data expand our understanding of the epidemiology of acute viral lower respiratory tract infections in Korean children and may be helpful to the clinicians and researchers interested in the control of viral respiratory tract infections.

Acute Disease↗

Virus isolations from patients in general practice, 1961-71.

During the period 1961-71 of 1785 viruses isolated from patients in the general population 503 (28%) were rhinoviruses, 465 (26%) influenza viruses, 248 (14%) enteroviruses, 234 (13%) herpes simplex virus, 132 (7%) parainfluenza viruses, 129 (7%) adenoviruses and 49 (3%) respiratory syncytial virus. Also isolated were 18 strains of mumps virus, 7 coronaviruses and 295 streptococci of groups A, C or G.Fluctuations were observed in the frequency with which respiratory syncytial virus, parainfluenza virus type 2, and the adenoviruses were isolated over the 10-year period.Influenza viruses types A and B, parainfluenza viruses types 1 and 2, respiratory syncytial virus, adenoviruses types 3, 4, 6, 7 and 21, and many enteroviruses were all associated with outbreaks.Infections with influenza viruses A and B and parainfluenza viruses types 1 and 2 came during the winter, whereas those with parainfluenza virus type 3, enteroviruses, and rhinoviruses were more frequently seen in the summer and early autumn.

Adenoviridae↗