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Virus antibodies in the serum of patients with liver disease.

Two hundred and twenty-six patients suspected of having liver disease were examined clinically, by a liver biopsy and laboratory test, according to a prospective scheme. Blood samples obtained just before the liver biopsy were coded and subsequently examined blindly, using the complement fixation test (CFT). The antigens were influenza A and B viruses, parainfluenza 1, 2, and 3, respiratory syncytial viruses, varicella, morbilli, cytomegalo and herpes simplex viruses. The sera were also examined by the CFT against Mycoplasma pneumoniae antigen. Antibodies against rubella virus were determined in a haemolysis-in-gel test. HBsAg and HBsAb were determined by a staphylococcal radioimmunoassay, and sera from patients with chronic active hepatitis (CAH) and chronic persistent hepatitis (CPH) were also examined for antibodies against hepatitis A virus by radioimmunoassay. Highly significant antibody titres against morbilli virus were found in patients with CAH and CPH. Patients with CPH or liver cirrhosis also had significantly higher titres against rubella virus than other groups. Some patients with liver granulomas had high titres against rubella virus. Only in one patient with CAH was a positive test for HBsAg found, and in one a positive test for HBsAb. Seven patients in the CAH and CPH groups had very high titres against both rubella and morbilli viruses.

Adolescent↗

The role of viruses in the aetiology of community-acquired pneumonia in adults.

BACKGROUND: The role of viruses in community-acquired pneumonia may have been previously underestimated. We aimed to study the incidence and clinical characteristics of community-acquired pneumonia (CAP) due to respiratory viruses in adults adding PCR to routine conventional laboratory tests. METHODS: Consecutive adult patients diagnosed of CAP from January 2003 to March 2004 were included. Conventional tests including cultures of blood, sputum, urine antigen detection of Streptococcus pneumoniae and Legionella pneumophila, and paired serologies were routinely performed. Nasopharyngeal swabs were processed for study of respiratory viruses through antigen detection by indirect immunofluorescence assay, isolation of viruses in cell culture and detection of nucleic acids by two independent multiplex RT-PCR assays. According to the aetiology, patients were categorized in 4 groups: group 1, only virus detected; group 2, only bacteria detected; group 3, viral and bacterial; and group 4, unkown aetiology. RESULTS: Of 340 patients diagnosed with CAP, 198 had nasopharyngeal swabs available and were included in this study. Aetiology was established in 112 (57%) patients: group 1, n=26 (13%); group 2, n=66 (33%); group 3, n=20 (10%). The most common aetiological agent was S. neumoniae (58 patients, 29%), followed by respiratory viruses (46 patients, 23%). Forty-eight respiratory viruses were identified: influenza virus A (n=16), respiratory syncytial virus A (n=5), adenovirus (n=8), parainfluenza viruses (n=5), enteroviruses (n=1), rhinoviruses (n=8) and coronavirus (n=5). There were two patients coinfected by two respiratory viruses. Serology detected 6 viruses, immunofluorescence 8, viral culture 12, and PCR 45. For the viruses that could be diagnosed with conventional methods, the sensitivity and specificity of RT-PCR was 85% and 92%, respectively. The only clinical characteristic that significantly distinguished viral from bacterial aetiology was a lower number of leukocytes (P=0.004). CONCLUSION: PCR revealed that viruses represent a common aetiology of CAP. There is an urgent need to reconsider routine laboratory tests for an adequate diagnosis of respiratory viruses, as clinical characteristics are unable to reliably distinguish viral from bacterial aetiology.

Aged↗

[Serological and epidemiological characteristics of acute pneumonias caused by Mycoplasma pneumoniae in Belgorod].

The etiology of the outbreaks of acute pneumonia in Belgorod was established and their epidemiological features were studied. The mycoplasmal etiology of all cases of acute pneumonia in children and adults, appearing alongside acute respiratory infections resulting from the preceding outbreaks caused by influenza viruses A/Prague/, B/Leningrad/369/75, and all types of parainfluenza viruses, was shown. The droplet mechanism of the transfer of infection was established, which was confirmed by the severity of the outbreak and a high rate of infection of the medical staff at hospitals for children and adults, where patients with mycoplasma-induced pneumonia were treated.

Acute Disease↗

[Vaccination of cattle against respiratory disease caused by virus infection (author's transl)].

Studies on the epizootiology of respiratory viruses in herds of cattle showed that infection by bovine respiratory syncytial (BRS) virus was a major cause of bovine respiratory disease in calves. Moreover, of IBR virus infection associated with respiratory symptoms were observed in the Netherlands since the Spring of 1974. Outbreaks of IBR virus infections occur throughout the Netherlands today. Initially, outbreaks ran a course marked by the typical symptoms of haemorhagic rhinitis, observed particularly in cows. Today, IBR virus infections have become enzootic on several farms, the majority of infections in young cattle being accompanied by respiratory symptoms that are identified as being due to IBR virus infection. Other virus infections that may cause respiratory disease, parainfluenza virus 3, adeno viruses and bovine virus diarrhoea (BVD) virus, were found to be enzootic in most herds in the Netherlands. However, there is no definite correlation between the occurrence of these infections and clinical symptoms. The use of vaccines against these virus infections is not advisable in the present situation. A vaccine against BRS virus infections is commerically available. Immunization by this vaccine against the pulmonary symptoms of BRS virus infection should be verified by a field trial. Unvaccinated control groups of calves should be included. In the experience of the present author, the immunity produced by this vaccine cannot prevent infection by the field strain of BRS virus. Postvaccination reactions were observed within seven days after vaccination with this vaccine in some cases. These were attributable to non-cytopathogenic strains of BVD virus, contaminating some batches of this commerical vaccine.

Animals↗

Investigations concerning the cultivation of myxo- and paramyxoviruses on chorioallantoic membrane fragments. Note I. Data on the multiplication of several myxo- and paramyxoviruses.

Influenza viruses A(H1N1) and A(H3N2) and parainfluenza viruses (Sendai mumps) were cultivated in chorioallantoic membrane (CAM) fragments maintained in media with different formulae, with or without daily medium changes, in roller or stationary tubes. Inoculation was performed either directly on CAM fragments in Petri dishes or by dilution of the virus-containing material in the medium. Infectant titers obtained in CAM fragments were similar to those recorded in embryonated eggs at 48 hours post inoculation (p.i.) in the case of influenza virus A(H1N1) and at 72 hours p.i. in that of Sendai and influenza A(H3N2) viruses; at 96 hours p.i. all the three viruses had titers superior to those found in the egg.

Animals↗

[Brain lesions in children with para-influenza].

An analysis of 1274 autopsy protocols of children parainfluenza was revealed as the main disease in 73 and as a concomitant disease in 107 cases, not infrequently as a generalized infection. In 13 observations a detailed study of the brain was performed. In 4 cases the involvement of the brain consisted mainly in cushion-like outgrowth of ependymocytes and endotheliocytes of the blood vessels. In 1 case a comprehensive analysis revealed isolated meningoencephalitis induced by type 3 parainfluenza virus. Besides, hemodynamic disorders and the nonspecific changes of moderate intensity were found. In 8 children only these nonspecific changes were detected.

Acute Disease↗

Quantitative method of measuring anti-viral activity of peritoneal dialysis effluent.

We have previously documented the presence of anti-viral activity in the peritoneal effluent of uremic patients treated by peritoneal dialysis. Anti-viral activity was determined by recording the inhibition of the cytopathic effect (CPE) of various viruses on Vero cells in a semi-quantitative manner based on observer's subjective judgment of the degree of inhibition of CPE. This degree of inhibition is a measure of the anti-viral activity. Although peritoneal dialysis effluent (PDE) was consistently seen to inhibit CPE we sought a quantitative method to accurately assess such anti-viral activity. To this end, the CPE of Parainfluenza virus (Para-3) on human kidney (Hu-K) cells was evaluated. The CPE of this virus is manifested by distinct plaques which can be easily counted after staining the tissue culture plate. Eighty-three (83) samples of PDE (0.5 ml) were added to Hu-K cells after which Para-3 was added. As control para-3 on Hu-K was used. The number of plaques obtained by virus control was taken as 100% CPE. The anti-viral activity of PDE was then recorded as the percentage of the CPE shown by virus control and averaged 20.9 +/- 28.6% (p less than 0.001 vs virus control). In addition PDE was diluted down to 1/64 concentration and the inhibitory effect again determined. Percent CPE of undiluted, 1/2, 1/64 PDE samples were 18.11 +/- 24.2, 33.57 +/- 18.16 and 77.93 +/- 17.4% respectively (p less than 0.01 between undiluted and 1/64 PDE). These data reaffirm that PDE possesses an inherent anti-viral activity which can now be quantitatively assessed using Para-3 on Hu-K cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗

Species-independent detection of RNA virus by representational difference analysis using non-ribosomal hexanucleotides for reverse transcription.

A method for the isolation of genomic fragments of RNA virus based on cDNA representational difference analysis (cDNA RDA) was developed. cDNA RDA has been applied for the subtraction of poly(A)(+) RNAs but not for poly(A)(-) RNAs, such as RNA virus genomes, owing to the vast quantity of ribosomal RNAs. We constructed primers for inefficient reverse transcription of ribosomal sequences based on the distribution analysis of hexanucleotide patterns in ribosomal RNA. The analysis revealed that distributions of hexanucleotide patterns in ribosomal RNA and virus genome were different. We constructed 96 hexanucleotides (non-ribosomal hexanucleotides) and used them as mixed primers for reverse transcription of cDNA RDA. A synchronous analysis of hexanucleotide patterns in known viral sequences showed that all the known genomic-size viral sequences include non-ribosomal hexanucleotides. In a model experiment, when non-ribosomal hexanucleotides were used as primers, in vitro transcribed plasmid RNA was efficiently reverse transcribed when compared with ribosomal RNA of rat cells. Using non-ribosomal primers, the cDNA fragments of severe acute respiratory syndrome coronavirus and bovine parainfluenza virus 3 were efficiently amplified by subtracting the cDNA amplicons derived from uninfected cells from those that were derived from virus-infected cells. The results suggest that cDNA RDA with non-ribosomal primers can be used for species-independent detection of viruses, including new viruses.

Animals↗

[Viral-microbial associations and the function of humoral factors of natural immunity in acute pneumonia patients].

A total of 359 patients with acute pneumonia and 152 practically healthy subjects comprising the control group were examined. Immunofluorescence was used to investigate nasopharyngeal washings for detecting antigens of influenza and parainfluenza viruses, respiratory-syncytial virus, adenoviruses, whereas serological studies according to the hemagglutination delay test with diagnostic agents for detecting influenza A1, A2, B, types 1, 2 and 3 parainfluenza, and the complement fixation test were made to detect antibodies against adenoviruses. Serological (65%) and immunofluorescence (63%) studies revealed associations of different viruses: type 3 and 1 parainfluenza, respiratory-syncytial virus (73%) with adenoviruses, influenza B, A2, type 2 parainfluenza. Association of different bacteria was observed in 67% of patients: hemolytic staphylococcus (65%), hemolytic streptococcus (50%), pneumococci (45%), P. aeruginosa (40%), P. mirabilis (35%), E. coli (30%), enterococci (25%). Associations of 3-2 causative agents were predominant (53%). Marked decrease in the content of complement and beta-lysins, elevation of the level of lysozyme were observed in patients with viral-bacterial and viral pneumonias as compared to the same characteristics in patients with bacterial pneumonia and in control group subjects.

Acute Disease↗

HVJ infection in alveolar macrophages of various laboratory animals.

Alveolar macrophages (AM) in bronchoalveolar lavage derived 6 species of laboratory animals (mouse, rat, Syrian hamster, Mongolian gerbil, guinea pig, and rabbit) were cultivated in vitro and were infected with HVJ, parainfluenza virus type 1, for the purpose of studying virus reproduction in these cells. Using infectivity assay, fluorescent antibodies and hemadsorption techniques, three categories were proposed depending on the potential of virus replication and antigen production. Category 1 (mouse, Syrian hamster, and guinea pig AM) was the most susceptible group to HVJ infection. All AM became antigen positive, and more than 10(6) PFU/ml of virus infectivity in the culture fluids was noted at 4 days after the virus infection. Rat and Mongolian gerbil AM were grouped into Category 2. Their AM showed a moderate virus yield (average 10(5) PFU/ml of infectivity) and antigen production. On the other hand, Category 3 (rabbit AM), exhibited no production of an infectious virus. It was likely that the virus penetrated into the AM but did not replicate complete virus. However, antigen positive cells ranging from 10 to 50% depended on the multiplicity of the infection to the AM. These findings suggest that HVJ infection in the rabbit AM was abortive (incomplete virus replication), since trypsin treatment of the culture fluids, which may contain incomplete HVJ particles, did not induce infectivity.

Animals↗

PIV-3 vaccine National Institutes of Health/Aviron.

Wyeth-Lederle, Aviron and NIH are developing a live intranasal parainfluenza virus (PIV) type 3 vaccine (cp45 vaccine). The vaccine is in phase II/III trials with Aviron [411386], and phase I trials with Wyeth-Lederle [408940]. It consists of a temperature-sensitive PIV-3 virus attenuated with an imported L gene mutation. This addition renders the virus 5-fold more restricted for replication in the respiratory tract of chimpanzees and also more temperature-sensitive [336999].

Administration, Intranasal↗

A functional antigenomic promoter for the paramyxovirus simian virus 5 requires proper spacing between an essential internal segment and the 3' terminus.

A previous analysis of naturally occurring defective interfering (DI) RNA genomes of the prototypic paramyxovirus simian virus 5 (SV5) indicated that 113 bases at the 3' terminus of the antigenome were sufficient to direct RNA encapsidation and replication. A nucleotide sequence alignment of the antigenomic 3'-terminal 113 bases of members of the Rubulavirus genus of the Paramyxoviridae family identified two regions of sequence identity: bases 1 to 19 at the 3' terminus (conserved region I [CRI]) and a more distal region consisting of antigenome bases 73 to 90 (CRII) that was contained within the 3' coding region of the L protein gene. To determine whether these regions of the antigenome were essential for SV5 RNA replication, a reverse genetics system was used to analyze the replication of copyback DI RNA analogs that contained a foreign gene (GL, encoding green fluorescence protein) flanked by 113 5'-terminal bases and various amounts of SV5 3'-terminal antigenomic sequences. Results from a deletion analysis showed that efficient encapsidation and replication of SV5-GL DI RNA analogs occurred when the 90 3'-terminal bases of the SV5 antigenomic RNA were retained, but replication was reduced approximately 5- to 14-fold in the case of truncated antigenomes that lacked the 3'-end CRII sequences. A chimeric copyback DI RNA containing the 3'-terminal 98 bases including the CRI and CRII sequences from the human parainfluenza virus type 2 (HPIV2) antigenome in place of the corresponding SV5 sequences was efficiently replicated by SV5 cDNA-derived components. However, replication was reduced approximately 20-fold for a truncated SV5-HPIV2 chimeric RNA that lacked the HPIV2 CRII sequences between antigenome bases 72 and 90. Progressive deletions of 6 to 18 bases in the region located between the SV5 antigenomic CRI and CRII segments (3'-end nucleotides 21 to 38) resulted in a approximately 25-fold decrease in SV5-GL RNA synthesis. Surprisingly, replication was restored to wild-type levels when these length alterations between CRI and CRII were corrected by replacing the deleted bases with nonviral sequences. Together, these data suggest that a functional SV5 antigenomic promoter requires proper spacing between an essential internal region and the 3' terminus. A model is presented for the structure of the 3' end of the SV5 antigenome which proposes that positioning of CRI and CRII along the same face of the helical nucleocapsid is an essential feature of a functional antigenomic promoter.

Animals↗

Hemagglutinin-neuraminidase of human parainfluenza 3: role of the neuraminidase in the viral life cycle.

The function of neuraminidase in the life cycle and pathogenesis of human parainfluenza virus type 3 (HPF3) was studied by analyzing a variant of HPF3 that has decreased neuraminidase enzymatic activity. The variant virus is more fusogenic than the wild-type virus during an acute infection. Cloning and sequencing of the fusion (F) and hemagglutinin-neuraminidase (HN) genes from this variant revealed a single amino acid change in the HN protein and no alterations in the F protein sequence. Analysis of the growth properties of this variant revealed a delay in release of virus particles into the supernatant. Addition of exogenous neuraminidase to the culture resulted in increased release of infections viral particles, suggesting that the viral neuraminidase is important for release of HPF3 from the infected cell surface. In addition, the behavior of the variant virus during high-multiplicity infection and in the presence of exogenous neuraminidase provided evidence that the neuraminidase of HPF3 determines the outcome of viral infection (cytopathic versus persistent) in cell culture.

Genetic Variation↗

Broad-spectrum antiviral activity of the IMP dehydrogenase inhibitor VX-497: a comparison with ribavirin and demonstration of antiviral additivity with alpha interferon.

The enzyme IMP dehydrogenase (IMPDH) catalyzes an essential step in the de novo biosynthesis of guanine nucleotides, namely, the conversion of IMP to XMP. The major event occurring in cells exposed to competitive IMPDH inhibitors such as ribavirin or uncompetitive inhibitors such as mycophenolic acid (MPA) is a depletion of the intracellular GTP and dGTP pools. Ribavirin is approved as an inhaled antiviral agent for treatment of respiratory syncytial virus (RSV) infection and orally, in combination with alpha interferon (IFN-alpha), for the treatment of chronic hepatitis C virus (HCV) infection. VX-497 is a potent, reversible uncompetitive IMPDH inhibitor which is structurally unrelated to other known IMPDH inhibitors. Studies were performed to compare VX-497 and ribavirin in terms of their cytotoxicities and their efficacies against a variety of viruses. They included DNA viruses (hepatitis B virus [HBV], human cytomegalovirus [HCMV], and herpes simplex virus type 1 [HSV-1]) and RNA viruses (respiratory syncytial virus [RSV], parainfluenza-3 virus, bovine viral diarrhea virus, Venezuelan equine encephalomyelitis virus [VEEV], dengue virus, yellow fever virus, coxsackie B3 virus, encephalomyocarditis virus [EMCV], and influenza A virus). VX-497 was 17- to 186-fold more potent than ribavirin against HBV, HCMV, RSV, HSV-1, parainfluenza-3 virus, EMCV, and VEEV infections in cultured cells. The therapeutic index of VX-497 was significantly better than that of ribavirin for HBV and HCMV (14- and 39-fold, respectively). Finally, the antiviral effect of VX-497 in combination with IFN-alpha was compared to that of ribavirin with IFN-alpha in the EMCV replication system. Both VX-497 and ribavirin demonstrated additivity when coapplied with IFN-alpha, with VX-497 again being the more potent in this combination. These data are supportive of the hypothesis that VX-497, like ribavirin, is a broad-spectrum antiviral agent.

Animals↗

Imidazo[1,2-a]-s-triazine nucleosides. Synthesis and antiviral activity of the N-bridgehead guanine, guanosine, and guanosine monophosphate analogues of imidazo[1,2-a]-s-triazine.

The first chemical synthesis of 2-aminoimidazo[1,2-a]-s-triazin-4-one (8), the corresponding nucleoside and nucleotide, and certain related derivatives of a new class of purine analogues containing a bridgehead nitrogen atom is described. Condensation of 2-amino-4-chloro-6-hydroxy-s-triazine (2) with aminoacetaldehyde dimethyl acetal followed by the ring annulation gave the guanine analogue 8. A similar ring annulation of 4-(2,2-dimethoxyethylamino)-s-triazine-2,6-dione (5) gave imidazo[1,2-a]-s-triazine-4,6-dione (9). Direct glycosylation of the trimethylsilyl derivative of 8 with 1-O-acetyl-2,3,5-tri-O-benzoyl-beta-D-ribofuranose in the presence of stannic chloride, followed by debenzoylation, gave the guanosine analogue 2-amino-8-(beta-D-ribofuranosyl)imidazo[1,2-a]-s-triazin-4-one (12b), which on deamination gave the xanthosine analogue 13. Phosphorylation of 12b gave 2-amino-8-(beta-D-ribofuranosyl)imidazo[1,2-a]-s-triazin-4-one 5'-monophosphate (II). The anomeric configuration has been determined unequivocally by using NMR of the 2',3'-O-isopropylidene derivate 10 and the site of ribosylation has been established by using 13C NMR spectroscopy. These compounds were tested against type 1 herpes, type 13 rhino, and type 3 parainfluenza viruses in tissue culture. Moderate rhinovirus activity was observed for several compounds at nontoxic dosage levels.

Antiviral Agents↗

Viral enhancement of nasal colonization with Haemophilus influenzae type b in the infant rat.

Infant rats infected with influenza A virus, Sendai (parainfluenza 1) virus or rat coronavirus were used to determine whether viral infection increases the intensity of nasal colonization with Haemophilus influenzae type b (HIB). Intranasal inoculation of HIB in rats previously infected with each of these viruses resulted in nasal HIB titers at least 100-fold higher than those for controls during the first 2 wk after HIB inoculation, and as much as 10,000-fold higher during the first week. Children with cough, sneezing, or rhinorrhea could be effective disseminators of HIB if they were as heavily and persistently colonized as these virus-infected animals.

Animals↗

The Seattle Virus Watch. VI. Observations of infections with and illness due to parainfluenza, mumps and respiratory syncytial viruses and Mycoplasma pneumoniae.

Seattle Virus Watch families were observed, 1965-1969, for infections with paramyxoviruses and M. pneumoniae by agent isolation and antibody assay of serial sera. Infection rates, based on serology, exceeded those in Tecumseh where families contained fewer young children. Rates per 100 person-years were 44.4 for parainfluenzavirus, 21.6 for respiratory syncytial (RS) virus and 12.3 for M. pneumoniae. Preschool children experienced the highest rates for RS and parainfluenza-viruses but, for the latter, rates were also high among older children and adults. Within invaded families infection rates generally varied inversely with age, although for M. pneumoniae the rates for adults and 6-9 year old children nearly equalled the infant rate. The introducers' identity and/or the age-specific infection rates in invaded families support the role of young schoolchildren in community spread of M. pneumoniae and, together with older children and adults, of RS virus. Young schoolchildren were less important than infants, preschoolers, and adults in spreading parainfluenza-viruses and less important than preschoolers and infants for mumps. The frequent infection of exposed older children and adults suggests that reinfection with all the agents studied is common. All agents spread significantly within families and secondary attack rates for the mostly non-immune infants indicated high infectivity of parainfluenza and mumps viruses. The basic high pathogenicity of these agents and of RS virus is indicated by the high frequency of illness among virus shedders (80-90%) and among seroconverting infants (greater than or equal 68%). The less frequent illness of older persons with serologically proven infection is consistent with diminished clinical response to reinfection. Parainfluenza-associated illnesses were relatively severe and contributed up to 9.3% to total respiratory illnesses. RS virus-related illnesses also were severe but contributed less (4-5%) to total respiratory disease. Mumps-associated illness was largely respiratory, 65% overall, 77% in infected infants and 75% above age 9. Thus, mumps virus emerges as another respiratory pathogen which is spread largely by 2-5-year old children rather than by schoolchildren with "typical" parotitis.

Adolescent↗

Nucleotide sequence analysis of the simian virus 41 gene encoding the large (L) protein and construction of a phylogenetic tree for the L proteins of paramyxoviruses.

The complete nucleotide sequence of the simian virus 41 (SV41) large (L) protein gene was determined. The L gene spanned 6883 nucleotides including a putative trailer RNA, and the L mRNA contained a single large open reading frame encoding a polypeptide of 2269 amino acids. Dot-matrix comparisons under stringent conditions identified domains highly conserved among paramyxoviruses. Domain 3 is the most highly conserved, and has been hypothesized to be the RNA polymerase active site. A phylogenetic tree was constructed from the sequences of the L proteins of seven paramyxoviruses. SV41 was most closely related to human parainfluenza virus type 2 (HPIV-2), and SV41, HPIV-2 and SV5 form a subgroup. The intergenic sequences at the nucleocapsid protein-phosphoprotein and haemagglutinin-neuraminidase-L protein gene junctions, and the 5' trailer sequence of SV41 were also determined, and it was shown that the first 13 nucleotides of the 5' trailer sequence are complementary to those of the 3' leader sequence. The intergenic, and gene-start and -end sequences of SV41, HPIV-2 and SV5 are shown.

Amino Acid Sequence↗