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Respiratory viruses and severe lower respiratory tract complications in hospitalized patients.

BACKGROUND: Acute respiratory viral infections are generally self-limited in healthy subjects but can lead to severe complications in immunocompromised hosts. We report the clinical impact of acute lower respiratory tract viral infections in hospitalized patients. MATERIALS AND METHODS: Of 1,001 fiberoptic bronchoscopies performed during a period of 5 years, 33 BAL samples were positive for respiratory viruses by cell culture. The main diagnosis, length of hospitalization, response to initial treatment, and the mortality rate at 30 days were analyzed. Spirometry performed before and after infection was compared in lung transplant recipients. RESULTS: The following respiratory viruses were identified in 33 cases: influenza A or B (n = 13), parainfluenza virus 1-3 (n = 7), rhinovirus (n = 5), respiratory syncytial virus (n = 4), and adenovirus (n = 4). All cases were immunocompromised patients who acquired new respiratory symptoms and/or radiologic abnormalities suggesting a pulmonary infection. Twenty-five patients (74%) did not respond to initial broad-spectrum antibiotics, and 11 patients (33%) required intensive care for respiratory failure. The overall mortality rate at 1 month was 24%. In patients with a sole viral pathogen identified in their BAL, the mortality rate was 39%. In lung transplant recipients (n = 10), the mean FEV(1) decreased from 2.2 to 1.9 L/s before and during the infection episode, respectively (p < 0.01); 3 months later, 60% of the patients had still not completely recovered to baseline values. CONCLUSION: Respiratory viruses recovered in BAL samples of immunocompromised patients are associated with severe lower respiratory complications. In lung transplant recipients, we observed a persisting impairment of pulmonary function.

Acute Disease↗

Etiology of acute lower respiratory tract infection in children at Srinagarind Hospital, Khon Kaen, Thailand.

We investigated the etiology of acute lower respiratory infection (ALRI) in children under 5 admitted to Srinagarind Hospital. The causative bacteria and viruses were determined by hemoculture and viral isolation from blood and nasopharyngeal aspirate samples. Antigens of respiratory syncytial virus (RSV) and Chlamydia trachomatis were detected using EIA. The 74 children less than 5 years of age with ALRI enrolled in our study were diagnosed with pneumonia (75.7%), croup (16.2%), and bronchiolitis (8.1%), respectively. Examination of blood or nasopharyngeal aspirate revealed viral or bacterial infections in 26 and 22 cases, respectively, whereas 5 of the children aged under 1 year (10%) were diagnosed with pneumonia caused by Chlamydia trachomatis. RSV was the most common virus detected (24.3%) and was associated with pneumonia and bronchiolitis, while the parainfluenza virus was the primary cause of croup. In cases of pneumonia, bacterial infections were identified in almost all of the cases: and Streptococcus pneumoniae and Haemophilus influenzae were the most commonly isolated (at 8.9% each). Mixed infections were detected in 8 cases (10.8%). The incidence of RSV infection peaked during the especially warm and cool seasons, whereas the bacterial infections were primarily associated with the relatively cool season. Our study indicates that a combined pneumococcal and Hib vaccine and a RSV vaccine would reduce the high rate of pneumonia in children under 5 years of age in Northeast Thailand.

Bacterial Infections↗

Identification of amino acids recognized by syncytium-inhibiting and neutralizing monoclonal antibodies to the human parainfluenza type 3 virus fusion protein.

Neutralizing monoclonal antibodies specific for the fusion (F) glycoprotein of human parainfluenza type 3 virus (PIV3) were used to select neutralization-resistant antigenic variants. Sequence analysis of the F genes of the variants indicated that their resistance to antibody binding, antibody-mediated neutralization or to both was a result of specific amino acid substitutions within the neutralization epitopes of the F1 and F2 subunits. Comparison of the locations of PIV3 neutralization epitopes with those of Newcastle disease and Sendai viruses indicated that the antigenic organization of the fusion proteins of paramyxoviruses is similar. Furthermore, some of the PIV3 epitopes recognized by syncytium-inhibiting monoclonal antibodies are located in an F1 cysteine cluster region which corresponds to an area of the measles virus F protein involved in fusion activity.

Amino Acids↗

Serological investigations for some bacterial and viral pathogens in fallow deer (Cervus dama) and wild boar (Sus scrofa) of the San Rossore Preserve, Tuscany, Italy.

Sera of 43 fallow deer (Cervus dama) of the San Rossore Preserve (Tuscany, Italy) were examined for antibodies against eight pathogens; one proved positive for Brucella sp., 21 for Listeria monocytogenes, 34 for Chlamydia psittaci, three for Coxiella burnetii, one for infectious bovine rhinotracheitis virus, 11 for parainfluenza-3 virus, 25 for bovine viral diarrhea virus and six for bovine respiratory syncytial virus. No age and sex difference in the positivity rates and titers was evidenced, while a sex difference was found both in rates of infection and in titers against parainfluenza-3 virus. Parainfluenza-3 infection was more prevalent in 1984 than in 1983 sampling. Sera of 20 wild boars (Sus scrofa) of the same preserve were examined for antibodies against five pathogens: four sera were positive for Brucella sp., while all were negative for Listeria monocytogenes, Chlamydia psittaci, Coxiella burnetii and Aujeszky's disease virus. Public and animal health involvement with these diseases are discussed for these respective host species.

Animals↗

Characterization of a live, attenuated human parainfluenza type 3 virus candidate vaccine strain.

Characterization of a temperature-sensitive and live, attenuated human parainfluenza type 3 virus strain (cp45) grown at a permissive temperature (32 degrees C) suggested that the virus efficiently multiplies in cell lines and retains antigenic and functional properties of the envelope glycoproteins. When grown at a nonpermissive temperature (39.5 degrees C), the cp45 virus exhibited poor replication; however, shifting to a permissive temperature allowed virus growth. Although at a nonpermissive temperature virus polypeptide synthesis was significantly reduced, the hemagglutinin-neuraminidase and fusion glycoproteins were transported to cell surfaces and retained their characteristic biologic activities. Studies on mRNA synthesis from the P protein gene suggested a poor transcriptional activity of the cp45 virus at a nonpermissive temperature. Results from this study indicate that the temperature sensitivity of cp45 virus is related to altered transcriptional activity and a marked reduction in virus polypeptide synthesis.

Antibodies, Monoclonal↗

The humoral immune response to recombinant nucleocapsid antigen of canine distemper virus in dogs vaccinated with attenuated distemper virus or DNA encoding the nucleocapsid of wild-type virus.

This study compared the humoral immune response against the nucleocapsid-(N) protein of canine distemper virus (CDV) of dogs vaccinated with a multivalent vaccine against parvo-, adeno-, and parainfluenza virus and leptospira combined with either the attenuated CDV Onderstepoort strain (n = 15) or an expression plasmid containing the N-gene of CDV (n = 30). The vaccinations were applied intramuscularly three times at 2-week intervals beginning at the age of 6 weeks. None of the pre-immune sera recognized the recombinant N-protein, confirming the lack of maternal antibodies at this age. Immunization with DNA vaccine for CDV resulted in positive serum N-specific IgG response. However, their IgG (and IgA) titres were lower than those of CDV-vaccinated dogs. Likewise, DNA-vaccinated dogs did not show an IgM peak. There was no increase in N-specific serum IgE titres in either group. Serum titres to the other multivalent vaccine components were similar in both groups.

Animals↗

Role of macrophages in virus-induced airway hyperresponsiveness and neuronal M2 muscarinic receptor dysfunction.

Viral infections exacerbate asthma. One of the pathways by which viruses trigger bronchoconstriction and hyperresponsiveness is by causing dysfunction of inhibitory M(2) muscarinic receptors on the airway parasympathetic nerves. These receptors normally limit acetylcholine (ACh) release from the parasympathetic nerves. Loss of M(2) receptor function increases ACh release, thereby increasing vagally mediated bronchoconstriction. Because viral infection causes an influx of macrophages into the lungs, we tested the role of macrophages in virus-induced airway hyperresponsiveness and M(2) receptor dysfunction. Guinea pigs infected with parainfluenza virus were hyperresponsive to electrical stimulation of the vagus nerves but not to intravenous ACh, indicating that hyperresponsiveness was due to increased release of ACh from the nerves. In addition, the muscarinic agonist pilocarpine no longer inhibited vagally induced bronchoconstriction, indicating M(2) receptor dysfunction. Treating animals with liposome-encapsulated dichloromethylene-diphosphonate depleted macrophages as assessed histologically. In these animals, viral infection did not cause airway hyperresponsiveness or M(2) receptor dysfunction. These data suggest that macrophages mediate virus-induced M(2) receptor dysfunction and airway hyperresponsiveness.

Animals↗

Duration of serologic response to five viral antigens in dogs.

OBJECTIVE: To determine whether vaccinated dogs either remained seropositive or responded serologically to revaccination for 5 key viral antigens after extended periods since their last vaccination. DESIGN: Serologic survey. ANIMALS: 322 healthy client-owned dogs. PROCEDURE: Dogs were > or = 2 years old and vaccinated against canine distemper virus (CDV), canine adenovirus-1 (CAV-1), canine adenovirus-2 (CAV-2), canine parainfluenza virus (CPIV), and canine parvovirus (CPV). On day 0, dogs were revaccinated with a vaccine from the same vaccine line as they had historically received. Antibody titers were measured in sera collected at day 0 (prevaccination titer) and 5 to 7 days later (postvaccination titer). Dogs were considered to have responded serologically if they had a day-0 serum neutralization titer to CDV > or = 1:32; a serum neutralization titer to CAV-1, CAV-2, or CPIV > or = 1:16; a hemagglutination inhibition titer to CPV > or = 1:80; or a > or = 4-fold increase in antibody titer after revaccination. RESULTS: The percentage of dogs that had titers at or greater than the threshold values or responded to revaccination with a > or = 4-fold increase in titer was 98.1% for CDV, 98.4% for CAV-1, 99.0% for CAV-2, 100% for CPIV, and 98.1% for CPV. CONCLUSIONS AND CLINICAL RELEVANCE: In most dogs, vaccination induced a response that lasted up to and beyond 48 months for all 5 antigens. Although not equivalent to challenge-of-immunity studies as a demonstration of efficacy, results suggest that revaccination with the same vaccine provides adequate protection even when given less frequently than the traditional 1-year interval. The study provides valuable information for clinicians to help determine appropriate revaccination intervals.

Animals↗

Respiratory virus disease in Malaysian children: a serological study.

Paired sera from 101 Malaysian children aged up to 10 years and suffering from respiratory illnesses were examined serologically for evidence of respiratory viral infections. Of these children, 32.6% showed rising antibody titres for one or more of the test agents. Respiratory syncytial virus appeared to be the main respiratory pathogen involved, followed by Mycoplasma pneumoniae, parainfluenza viruses, adenoviruses, and influenza A virus. These findings are generally similar to those reported by others in temperate and tropical countries.

Antibodies, Viral↗

Giant cell arteritis, polymyalgia rheumatica, and viral hypotheses: a multicenter, prospective case-control study. Groupe de Recherche sur l'Artérite à Cellules Géantes.

OBJECTIVE: Although suspected, a viral etiology has never been proven in giant cell arteritis (GCA). We tested for viruses known to induce multinucleated giant cells in human pathology, which include the parainfluenza viruses (HPIV), respiratory syncytial virus, measles virus, herpesviruses type 1 and 2, and the Epstein-Barr virus. METHODS: A multicenter case-control study on incident cases of temporal arteritis (TA) and polymyalgia rheumatica (PMR). Population based age and sex matched controls were randomly selected. Serological tests for IgG and IgM directed against the viruses listed above were performed, on blood samples taken at the time of clinical diagnosis. RESULTS: Three hundred five new patients were included over a 5 year period, of whom 159 presented with positive biopsy TA, 70 with negative biopsy TA, and 76 with negative biopsy PMR. Thirty-eight percent of cases versus 20.9% of controls were positive for IgM directed against HPIV (p = 0.00005). The association was stronger in the positive TA subgroup [positivity rate 43.31%; odds ratio with controls 2.89 (95% CI 1.82-4.60, p = 0.000006)] than in the PMR or negative biopsy TA subgroups. Only HPIV type 1 was associated with the disease, regardless of the season or the geographical origin of the cases. Positivity rates for HPIV types 2 and 3 and for the other viruses tested were similar in cases and controls. CONCLUSION: Our findings suggest that reinfection with HPIV type 1 is associated with the onset of GCA in a subset of patients, particularly in cases with positive TA biopsy.

Aged↗

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↗

Primary cytotoxic response of bovine peripheral blood leukocytes to parainfluenza type 3 virus infection.

Peripheral blood mononuclear cells were isolated from calves following Parainfluenza Type-3 (PI-3) virus infection and incubated with autologous and non-autologous PI-3 virus-infected muscle cells in a 4 h chromium release assay. Peaks of cytotoxicity, which ranged from 12% to 53% were observed between Days 6 and 9 post infection, with an effector to target cell ratio of 100:1. Killing of PI-3 virus-infected or uninfected mismatched muscle cells was never more than 5%.

Animals↗

Sequence determination of the mumps virus HN gene.

The hemagglutinin-neuraminidase protein (HN) of mumps virus was purified by immunoaffinity chromatography and fragmented by the combined action of CNBr and trypsin. The resulting peptides were separated by HPLC and sequenced by automated Edman degradation. Using this HN-specific amino acid sequence data, a degenerate oligonucleotide was produced and subsequently used to screen a mumps virus cDNA library to isolate HN-specific clones. The complete nucleotide sequence of the HN gene was determined. The monocistronic HN mRNA is approximately 1900 nucleotides long and encodes a single open reading frame of 582 amino acids. The HN protein has a unique hydrophobic stretch of 19 amino acids at its N-terminus that apparently anchors the protein in the viral envelope. A comparison of the mumps virus HN protein sequence with the sequences of the other known paramyxovirus HNs indicates that mumps virus is most closely related to SV-5, followed in decreasing order by NDV, parainfluenza virus 3, and Sendai virus.

Amino Acid Sequence↗

Viral respiratory infection in Curitiba, Southern Brazil.

OBJECTIVE: Viral infection is the major cause of respiratory tract pathology affecting individuals of all ages, especially children and immunocompromised patients. There is a paucity of data on etiological and epidemiological infection caused by virus, in Southern Brazil. The aim of this study was to evaluate the seasonality, and the incidence of the viruses involved in respiratory infections in Curitiba, South Brazil. METHODS: Two hundred seventy-three nasopharyngeal aspirates (NPA) from primary care health units and 1348 NPA or bronchoalveolar lavages from a tertiary-care teaching hospital were studied. Viruses were identified by indirect immunofluorescence and cellular culture. RESULTS: Thirty percent of tested samples scored positive for the presence of virions. The percentages of infection for each virus from outpatients and hospitalized, respectively, were 3.3, 19.3 for respiratory syncytial virus; 13.9, 3.5 for influenza A, 3.0, 1.7 for influenza B; 3.3, 5.6 for parainfluenza virus and 2.2, 0.8 for adenovirus. A higher incidence of positive results was found during the winter season, thus showing a pattern of seasonality. CONCLUSION: Viral agents are one of the main etiologies of respiratory tract pathology in the population studied.

Adolescent↗

Rapid detection of influenza A and B viruses in clinical specimens by Light Cycler real time RT-PCR.

BACKGROUND: the influenza viruses cause morbidity and mortality annually among children and elderly. Surveillance and rapid diagnosis is imperative in the reference laboratory, as clinical symptoms are insufficient for proper diagnosis. OBJECTIVES: this study involved the design of a rapid detection method for influenza A and B viruses using real time RT-PCR from clinical specimens. Methods were specifically designed for use on the Light Cycler. The sensitivity and specificity were also to be determined. STUDY DESIGN: the identification and discrimination of influenza A and B viruses employs two dual probe systems based on fluorescence resonance energy transfer (FRET) technology. Following submission by physicians participating in the Florida sentinel influenza network, 58 specimens were chosen for testing using both tissue culture and Light Cycler methods. RESULTS: of the 35 identified positive for influenza virus via tissue culture isolation, the Light Cycler results matched identification and typing with 100% agreement. However, the Light Cycler recognized 16 additional specimens that were positive for the presence of the virus. RT-PCR and nucleotide sequencing confirmed the presence of influenza A virus in these specimens. Using tenfold serial dilutions, the sensitivity of the Light Cycler method was determined to be 0.01 TCID50. The lower limit of RNA detection was determined as 1.6 x 10(-7) microg for influenza A virus, and 1.2 x 10(-7) microg for influenza B virus. Specificity of the Light Cycler method was determined by testing specimens containing adenovirus, parainfluenza virus and echovirus, all of which yielded negative results with no discernible background. CONCLUSIONS: overall, this newly developed method of simultaneous detection and typing of influenza types A and B using the Light Cycler proves to be more sensitive than tissue culture isolation, with corresponding specificity. This technique may be valuable for surveillance and rapid identification of influenza for early diagnosis.

Humans↗

The classification of viruses infecting the respiratory tract.

Following the boom in respiratory virology in the 1960s, species of rhinoviruses, coronaviruses, enteroviruses, adenoviruses, parainfluenza viruses and respiratory syncytial virus were added to influenza and measles viruses as causes of respiratory tract infection. In restricted patient groups, such as the immunocompromised, members of the family of herpesviruses including herpes simplex, cytomegalovirus, varicella-zoster virus, Epstein-Barr virus and human herpes virus 6 have also been associated with respiratory disease. This list of pathogens was extended last year with the discovery of a novel virus, the human metapneumovirus. More than 200 antigenically distinct viruses have been documented as causes of sporadic or epidemic respiratory infections in infants, children and adults. However, this varied and diverse group can be divided among six distinct families. Understanding some of the basic biology of these families gives an insight into possible strategies for diagnosis, control and therapy.

Adenoviridae↗

Viral infections in childhood leukemia.

The importance of viral infections in 150 children receiving therapy for leukemia was studied prospectively by application of comprehensive viral diagnostic procedures. One hundred five viral infections were identified, with herpes simplex virus and varicella-zoster virus being the most common agents. The spectrum of viruses associated with serious illness was wider than that in previous studies, and adenoviruses, parainfluenza viruses, rhinoviruses, and enteroviruses were important causes of morbidity. Viral isolation was the most sensitive diagnostic procedure used because complement fixation serology was falsely negative in two-thirds of cases. The occurrence of viral infection may be a previously overlooked important cause of respiratory tract infection and acute pyrexia of unknown origin. Viral infection rates were highest in patients with acute myeloblastic leukemia, in induction and relapse, and in patients treated with newer chemotherapeutic schedules. Thus, viruses are important pathogens in children with leukemia.

Adolescent↗