Abstract Little is known about a) the differences in allergic and respiratory diseases between the Finnish and Russian populations, and the environmental factors associated with those differences, and b) exposure to pollen grains indoors and the efficiency of penetration of pollen from outdoor to indoor air. This thesis is based on a cross-sectional population-based epidemiological study conducted in Imatra (Finland) and Svetogorsk (Russia) in 2003 and a rotorod-type-sampler-based pollen study conducted in the province of South Karelia (Finland) between 2003 and 2004. The prevalence of allergic diseases was higher among Finnish than Russian schoolchildren. The symptoms among allergic children were more severe, and the occurrence of respiratory infections was in general more frequent in Russia than in Finland. In the logistic regression analyses the risk of asthma was particularly related to high maternal smoking exposure, and the risk of the common cold was related to high combined parental smoking during infancy (adjusted OR 1.83, 95% CI 1.06–3.17) in Finnish children. Among Russian children, allergic conjunctivitis was related to maternal smoking, while the common cold was inversely related to paternal and parental smoking (0.60, 0.37–0.98 and 0.31, 0.11–0.83, respectively) during the study period. The risk of asthma was inversely related to any indoor dog-keeping in Finland (0.35, 0.13–0.95), whereas in Russia the risk of asthma was increased in relation to combined indoor cat exposure during infancy and the study period (4.56, 1.10–18.91). The concentrations of pollen grains decreased from abundant (0–855 pollen grains per cubic meter, pg/m3) to low (0–3 pg/m3), when moving from outdoors to indoors and further. The differences in diseases and symptoms in these two closely related populations could be ascribed to differences in culture, exposures, diagnostic criteria and treatment. The concentrations of pollen in indoor air during the flowering period were mostly on a level high enough to cause reactions in only the most sensitive subjects. The results suggest that more efforts should be directed to reducing parental smoking, to studying the role and effects of nationally different animal exposures in childhood, and to assessing the importance of different penetration routes of pollen grains. / Tiivistelmä Suomen ja Venäjän välisistä allergioiden ja hengitystietulehdusten esiintymiseroista ja esiintymiseen vaikuttavista ympäristötekijöistä tiedetään varsin vähän. Myös tutkimuksia siitepölyille altistumisesta sisätiloissa ja siitepölyjen tunkeutumiskyvystä ulkoilmasta sisäilmaan on niukasti. Tutkimus yhdistää sekä lääketieteellisen että luonnontieteellisen tutkimusalan tutkimustraditiot sekä atooppisten sairauksien ja/tai hengitystietulehdusten tärkeimpien määrittäjien tarkastelun yhdeksi tutkimuskokonaisuudeksi. Väestö- ja kyselylomakepohjainen poikkileikkaustutkimus toteutettiin Suomen ja Venäjän rajan molemmin puolin sijaitsevissa Imatran ja Svetogorskin kaupungeissa vuonna 2003. Tutkimusväestö koostui 512 suomalaisesta ja 581 venäläisestä 7–16-vuotiaasta koululaisesta (osallistumisaste 79 %). Rotorod-tyyppisen keräimen käyttöön perustuva siitepölytutkimus toteutettiin erilaisissa ulko- ja sisätiloissa Lappeenrannan ja Imatran kaupungeissa, Rautjärven kunnassa ja valtatie 6:lla vuosina 2003 ja 2004. Atooppisten sairauksien esiintyvyys oli runsaampaa suomalaisten koululaisten keskuudessa. Sitä vastoin allergisten lasten kokemat oireet olivat voimakkaampia ja hengitystietulehdusten esiintyvyys oli runsaampaa venäläisten koululaisten keskuudessa. Astmariski kytkeytyi erityisesti äidin runsaalle tupakoinnille altistumiseen raskauden (vakioitu OR 3.51, 95 % luottamusväli 1.00–12.3), ensimmäisen elinvuoden (3.34, 1.23–9.07) ja tutkimuksen aikana (3.27, 1.26–8.48). Nuhakuumeen riski oli suurentunut suomalaisten koululaisten keskuudessa, jotka olivat altistuneet molempien vanhempien runsaalle tupakoinnille ensimmäisen elinvuoden aikana (1.83, 1.06–3.17). Äidin tupakoinnille ensimmäisen elinvuoden (4.53, 1.49–13.8) ja tutkimuksen aikana (2.82, 1.07–7.44) altistuneilla venäläisillä oli suurentunut allergisen silmän sidekalvotulehduksen riski. Tutkimuksen aikainen isän ja vanhempien tupakointi vähensi nuhakuumeen riskiä (0.60, 0.37–0.98; 0.31, 0.11–0.83) Venäjällä. Suomessa koiranpito sisätiloissa vähensi astmariskiä (0.35, 0.13–0.95), vastaavasti Venäjällä raskauden jälkeinen sisätiloissa tapahtuva kissa-altistus lisäsi koululaisten astmariskiä (4.56, 1.10–18.91). Siitepölyjen pitoisuudet pienenivät siirryttäessä ulkoa (0–855 siitepölyhiukkasta ilmakuutiossa; sp/m3) sisätiloihin (0–17 sp/m3). Ympäristöaltisteisiin ja sairauden ennusteeseen vaikuttavat sekä kansallinen kulttuuri ja vakiintuneet tavat, että erot diagnosointikriteereissä, yleisessä tautitietoisuudessa ja lääkkeiden saatavuudessa. Näin ollen altisteiden voimakkuus ja kesto sekä terveysvaikutukset voivat vaihdella merkittävästi lähellä toisiaan sijaitsevien alueiden välillä. Siitepölypitoisuudet sisätiloissa olivat pääosin tasolla, jolle altistuminen aiheuttaa oireita vain kaikkein herkimmille allergisille. Tutkimuksen tulosten mukaan lisää voimavaroja tulisi suunnata passiiviselle tupakoinnille altistumisen vähentämiseen erityisesti yksilökehityksellisesti herkkien varhaisvaiheiden aikana, kansallisten eläinaltistuserojen terveysvaikutusten selvittämiseen sekä siitepölyjen erilaisten kulkeutumisreittien merkityksen tutkimiseen.

A reação em cadeia por polimerase (PCR) e suas variantes tem sido, desde o isolamento do metapneumovírus humano (hMPV), a técnica mais utilizada para a detecção do vírus em secreções respiratórias de diferentes grupos de pacientes. Entretanto, a interpretação de estudos abordando aspectos epidemiológicos e patogenéticos da infecção pelo hMPV tem sido dificultada pelo uso de uma grande variedade de técnicas de PCR \"in house\" na ausência de uma técnica \"padrão ouro\" claramente definida. A avaliação da sensibilidade, especificidade e reprodutibilidade de qualquer técnica molecular \"in house\" é um passo crucial para podermos comparar estudos realizados por diferentes grupos de pesquisa e diferentes grupos de pacientes. Este estudo teve como objetivos a padronização de duas técnicas de PCR - convencional e em tempo real - para a detecção do hMPV em secreções respiratórias e a avaliação da concordância existente entre as técnicas. Entre 228 amostras de lavado de nasofaringe coletadas de receptores de transplante de células tronco hematopoiéticas com sintomas de infecção respiratória aguda, 10 (4,4%) foram positivas para a presença do hMPV pela técnica de PCR convencional enquanto que 11 (4,8%) foram positivas pela técnica de PCR em tempo real. A concordância entre as técnicas, medida pelo índice Kappa para um intervalo de confiança de 95%, foi de 0,95, ou seja, quase perfeita. / The polymerase chain reaction (PCR) has been, since the isolation of the virus in 2001, the most used technique for detection of human metapneumovirus (hMPV) in respiratory specimens of several groups of patients. However, the interpretation of studies regarding the epidemiology and pathogenesis of hMPV infection has been hindered by the use of a great variety of PCRs techniques for hMPV detection, in the absence of a clearly defined \"gold standard\". The assessment of the sensitivity, specificity and reproducibility of any in-house molecular technique is a crucial step to allow the comparison of studies conducted in different settings and different groups of patients. The aim of the present study was to standardize two in-house PCR assays a conventional PCR and a real-time PCR for detecting hMPV in nasopharyngeal aspirates and to evaluate the agreement between the two assays. Of 228 samples of nasopharyngeal aspirates obtained from hematopoietic stem cell transplant recipients with acute respiratory symptoms, 10 (4.4%) were positive for hMPV by conventional PCR whereas 11 (4.8%) were positive by real-Time PCR. The agreement of both assays, measured by Kappa Index, was almost perfect (0.95, 95% confidence interval).

Introducción: Las infecciones agudas del tracto respiratorio (IRA) son muy prevalentes, las IRAs bajas constituyen la cuarta causa de muerte a nivel mundial. Los agentes causales más comunes en niños son el Rinovirus (RV) y el Virus sincitial respiratorio (VSR). Existen tres especies de RV (A, B, C), estudios recientes han demostrado que la sintomatología y severidad de la enfermedad varía dependiendo de la especie de RV por la que hayan sido infectados los pacientes. Objetivo: Evaluar la prevalencia de Rinovirus en muestras de hisopado nasofaríngeo de niños con diagnóstico clínico de IRA en Lima, Perú durante el periodo 2009-2010. Materiales y métodos: Estudio retrospectivo de muestras de hisopado nasofaríngeo en niños, procesadas por la técnica RT-PCR para identificación de RV y sus especies. La población está compuesta por pacientes pediátricos en el Hospital Nacional Cayetano Heredia. Se analizó las variables por medio de la prueba de Chi cuadrado y Fischer. Resultados: RVA se detectó en 10.26%, RVB en 16.67%, RVC 73.9%. Grupo etario más prevalente fue de 0-5 meses. Signos y síntomas más comunes fueron tos, fiebre, rinorrea y dificultad respiratoria. Se encontró asociación entre sibilancias y RVA; tos, sibilancias e inyección conjuntival con RVC. Se halló pico de casos por RVC durante marzo, junio y noviembre. Conclusión: Se encontró alta prevalencia de infección por RVC en pacientes pediátricos, principalmente en pacientes de 0-5 meses. Distribución mensual muestra aumento de casos en marzo y junio. Se sugiere realizar vigilancia epidemiológica y estudios longitudinales para el estudio de este patógeno. / Introduction: Acute respiratory tract infections (ARTI) are a very prevalent group of diseases, lower ARTI represent the fourth cause of death worldwide. In children, the two most usual agents are Rhinovirus (RV) and Syncytial respiratory virus (SRV). RV is responsible for most is related with lower respiratory tract infections. Scientists have identified three RV species (A, B, C), recent studies have reported that symptomatology and severity vary within RV species. Objective: Asses the prevalence of Rhinovirus on nasopharyngeal swab samples of children with clinical diagnosis of ARTI in Lima, Peru during 2009-2010. Materials and method: Retrospective study about nasopharyngeal swab on children, which were processed through RT-PCR technique to identify RV and its species. The study population was pediatric patients, with clinical diagnosis of ARTI, at Hospital Nacional Cayetano Heredia. This investigation project will be revised by the ethics committee from Universidad Peruana de Ciencias Aplicadas. Results: RVA was detected in 10.26% of cases, RVB 16-67% and RVC in 73.9%. The most prevalent age group was the 0-5 months old. The most common signs and symptoms were cough, fever, rhinorrhea, and respiratory distress. The study found association between wheezing and RVA infection, cough, wheezing and conjunctival injection and RVC infection. There was a peak in RVC cases during the March, June, and November. Conclusion: We found a high prevalence for RVC infection, mainly in children between 0-5 months old. Monthly distribution showed an increase of RVC cases during March and June; epidemiological surveillance and longitudinal studies should be encouraged. / Tesis

Despite advances in health promotion through efforts to reduce tobacco smoking, tobacco-related health conditions have continued to be significant. Exposure to secondhand smoke has been identified as a health risk also in addition to infant health risks related to maternal smoking. In contrast, breastfeeding has been found to promote infant health and is strongly encouraged. Despite literature supporting both of these statements, the combined effects of both breastfeeding and maternal smoking on infant wellbeing have not been delineated. Otitis media represents a common health problem among infants and young children. Tobacco exposure has been shown to increase its incidence while breastfeeding has been shown to reduce its occurrence. In the current study, a consecutive sample of all infants less than 5 years of age with otitis media and breastfed for at least 6 months was collected from a busy urban clinic for analysis. A survey tool was administered to those meeting study criteria. Primary analysis examined the odds ratio of developing otitis media among breastfed infants between those whose mothers smoked tobacco and those whose mothers did not. As a result, the association between the protective effects of breastfeeding and the detrimental effects of maternal smoking was evaluated in relation to the development of otitis media. Secondary variables including demographics, family history, past medical and birth history, and secondhand smoke exposure were also assessed. Results failed to demonstrate a significant difference in otitis media between the 2 cohorts in this study, and of the secondary variables, only cranio-facial deformities and/or a family history of these conditions resulted in higher otitis media occurrence. Further study with larger populations with higher tobacco use rates may offer additional insights into this matter.

No description available.

<p><strong>Background</strong>: Human rhinovirus/enterovirus (HRV/ENT) infections are commonly identified in children with acute respiratory infections (ARIs), but data on their clinical severity remains limited. We compared the clinical severity of HRV/ENT to respiratory syncytial virus (RSV), influenza A/B (FLU) and other common respiratory virus in children.</p> <p><strong>Methods</strong>: Retrospective study of children with ARIs and confirmed single positive viral infections on mid-turbinate swabs by molecular assays. Outcome measures included hospital admission and, for inpatients, a composite end-point consisting of intensive care admission, hospitalization greater than 5 days, oxygen requirements or death.</p> <p><strong>Results</strong>: A total of 116 HRV/ENT, 102 RSV, 99 FLU and 64 other common respiratory viruses were identified. Children with single HRV/ENT infections presented with significantly higher rates of underlying immunosuppressive conditions compared to those with RSV (37.9% vs 13.6%; p</p> <p><strong>Conclusions</strong>: Children with HRV/ENT had a more severe clinical course than those with RSV and FLUA/B infections and often had significant comorbidities. These findings emphasize the importance of considering HRV/ENT infection in children presenting with severe acute respiratory tract infections.</p> / Master of Science (MSc)

Le genre Entérovirus (EV) (famille des picornaviridae) est composé de petits virus à ARN non enveloppées classés en 12 espèces dont 7 sont pathogènes pour l'homme : 4 espèces (A-D) d'enterovirus humains (HEV) et trois espèces (A-C) de rhinovirus humains (HRV). Dans le genre enterovirus, les HRV et HEV sont reconnus comme des pathogènes respiratoires fréquemment responsables d'infections des voies aériennes supérieures et inférieures chez l'enfant et l'adulte. Entre 2009 et 2012, de nouveaux génotypes d'HEV à tropisme respiratoire (HEV-68, 104, 109, et le CVA-21) ont été décrits dans des cas isolés ou épidémiques démontrant la capacité des espèces A à D à induire des infections respiratoires basses humaines.La première phase de ce travail de thèse a eu pour objectifs de préciser le rôle étiologique des infections à EVs; d'identifier les génotypes potentiellement responsables des pathologies respiratoires pédiatriques nécessitant une hospitalisation, mais aussi d'analyser et de comparer les caractéristiques cliniques et épidémiologiques entre les différents groupes de génotypes identifiés. Nous avons réalisé une étude rétrospective sur une cohorte de 309 enfants hospitalisés au CHU de Reims entre septembre 2009 et juin 2010 pour une infection respiratoire aiguë non documentée microbiologiquement par la réalisation des tests virologiques et bactériologiques conventionnels. Nos résultats montrent que le génome des EVs (HEV et HRV) est retrouvé dans 60,5% (187/309) des aspirations naso-pharyngées des enfants hospitalisés, distinguant 15 infections à HEV (dont 10 souches HEV-68) et 172 à HRV. Les cas de bronchiolite et d'exacerbation de l'asthme (133/187) positifs pour la détection des souches HEV (12/133) étaient plus âgés (P=0,003) et plus fréquemment associés avec une détresse respiratoire (P=0,01) et un besoin en oxygénothérapie au moment de leur hospitalisation (P=0,01) que les cas infectés par un HRV. De plus, nous avons mis en évidence pour la première fois en France la circulation épidémique de souches d'HEV-68 (10/15 des souches d'HEV détectées) isolées au cours de l'automne 2009 chez des enfants hospitalisés pour une infection respiratoire aiguë. Nos résultats fournissent de nouvelles informations sur ce génotype ré-émergent qui semble présenter un tropisme respiratoire spécifique des voies respiratoires inférieures.La seconde phase de ce travail de thèse s'est intéressée à étudier les mécanismes liés au développent des processus inflammatoires de la muqueuse au cours de l'infection des voies respiratoires basses par HEV. A l'aide d'un modèle in vitro de cellules respiratoires humaines (A549) infectées par HEV-B (CVB5, Mitchell), nous avons observé que l'infection réplicative des HEV dans les cellules A549 induisait une augmentation dose et temps-dépendante des ARNm, et des protéines IL-8, MCP-1 et RANTES.En conclusion, nos résultats obtenus à partir de prélèvements respiratoires dans le cadre de notre étude de cohorte suggèrent que les EVs représentent une cause étiologique fréquente d'infections respiratoires basses chez l'enfant avec une pathogénicité supérieure des HEVs (principalement dans notre étude HEV-68) par rapport aux souches HRVs. De plus, nos résultats obtenus à partir d'expérimentation in vitro démontrent que les HEVs du groupe B sont capables d'induire au cours de l'infection des cellules épithéliales alvéolaires humaines (A459) une sécrétion spécifique d'IL-8, MCP-1 et RANTES. La production de ces chimiokines correspond à une réponse innée de la cellule épithéliale humaine infectée par les HEVs: nous avons montré pour la première fois que ce mécanisme était en partie régulé par l'activation de la voie non canonique de NF-kB via la protéine NIK dans la cellule épithéliale respiratoire humaine. / The Enterovirus (EV) genus (picornaviridae family) consists of small non-enveloped positive RNA viruses classified in 12 species of which 7 are pathogenic for humans: four species (A-D) of human enterovirus (HEV) and three species (A-C) of human rhinovirus (HRV). Among the EV genus, HEV and HRV are recognized as leading causes of acute respiratory tract infections (ARTIs) in human. Between 2009 and 2012, new HEV respiratory genotypes (e. g. HEV-68, 104, 109, 117 and CVA-21) have been described in isolated cases or outbreaks supporting the ability HEV species A to D to induce lower respiratory tract infections. This supports the hypothesis of an underestimation of the prevalence and etiological role of EVs in pediatric acute respiratory tract infections (ARTIs) (more specifically bronchitis, bronchiolitis and asthma exacerbation).To assess the etiological role and the clinical characteristics of HRV and HEV infections in pediatric patients hospitalized for ARTIs, we conducted a retrospective study of 309 hospitalized pediatric patients in University Hospital Centre of Reims with microbiologically unexplained ARTIs from September 2009 to June 2010. Among the 309 ARTIs, 15 HEV and 172 HRV strains were identified. Among bronchiolitis and asthma exacerbation cases (n=133), HEV infected cases were older (P=0.003) and were more frequently associated with a respiratory distress (P=0.01) and a need for oxygen therapy at the time of admission (P=0.01) than cases infected by HRV strains. Interestingly, during this retrospective study, we provided evidence that during the fall 2009 in France, HEV-D68 strains were responsible for a low proportion of pediatric cases hospitalized for acute airway diseases including bronchiolitis and asthma exacerbation.To identify the mechanisms that can regulate the development of airway mucosa inflammation during HEV respiratory lower tract infections, we investigated the production of chemokines by HEV infected human alveolar epithelial cells (A549). Using in vitro model A549 cells infected by HEV-B (CVB5, Mitchell), we demonstrated that HEV-B strains isolated from upper respiratory tract of child with bronchiolitis could actively replicate in various human airway epithelial cells, and that this replicative infection induced specific dose and time-dependent increases in mRNA and protein secretion of IL-8, MCP-1 and RANTES, but not of all other CC and CXC human chemokines tested. The protein secretion of these chemokines appeared to be significantly increased at 48 and 72 hours post-infection in culture treated by low-doses of IFN-γ in comparison with mock-infected cells (P <0.001), and was correlated to the viral replication activity. In second time, we explore the pathogenic mechanisms that can regulate inflammatory responses to HEV in lower respiratory airways. We show that HEV infection induced a time-dependent increase of NIK protein accumulation that peaks at 16 hours post-infection (H P-I). NIK protein accumulation mediated the processing of p100 in p52, which association with Rel B was evidenced in nuclear compartment between 16 and 48 H P-I.In conclusion, our findings indicate that EVs are a common cause of lower respiratory tract infections in pediatric patients with a potential higher pathogenicity of HEV strains (mainly HEV-68) by comparison to HRV strains. Moreover, our in vitro results demonstrated that HEV are capable to induce the release of specific chemokines (IL -8, MCP-1 and RANTES) by alveolar epithelial cells during a replicative infection. Finally, we demonstrated for the first time that this innate airway epithelial cell response against HEV infection was partly regulated by the activation of the non-canonical NF-kB via NIK protein.

O bocavirus humano (HBoV) é um parvovirus recentemente identificado em associação com a presença de sintomas de infecção do trato respiratório. Esse vírus possui um genoma de aproximadamente 5217 nucleotídeos que contém 3 open reading frames que codificam 4 proteínas (NS1, NP-1, VP-1 e VP-2). HBoV tem sido detectado em amostras respiratórias de diversas partes do mundo, incluindo Austrália, América do Norte, Europa, Ásia e África, o que sugere uma distribuição global desse vírus. Entretanto, nenhum estudo longitudinal de HBoV em amostras respiratórias foi realizado na América Latina. Dessa forma, nós realizamos um estudo prospectivo de HBoV em lavados nasofaríngeos (LFNs) coletados de pacientes com sintomas de infecção do trato respiratório (IRA) atendidos em um hospital universitário de Ribeirão Preto, SP e em um hospital universitário de Salvador, BA no período entre 2005 a 2007. 1288 LFNs de 1217 pacientes foram encaminhados ao laboratório de virologia e foram testados por PCR para HBoV. Desses pacientes, 962 eram menores de 5 anos e 177 eram maiores de 5 anos. Além disso, também foram analisados 50 LFNs de crianças menores de 5 anos que não tinham sintomas respiratórios. Todas as amostras positivas para HBoV foram testadas para todos os outros vírus respiratórios, incluindo o vírus sincicial respiratório (HRSV), rinovirus humano (HRV), influenza humano (HFLU), metapneumovirus humano (HMPV), parainfluenza humano (HPIV), coronavirus humano (HCoV) e adenovirus humano (HAdV). A carga viral de HBoV foi determinada por PCR em tempo real em todas as amostras positivas e o genoma completo de 19 amostras de HBoV foi seqüenciado. Com intuito, de fazer um levantamento sorológico e determinar sítios replicativos de HBoV, nós ainda clonamos e expressamos em S. cerevisae (Y258) o gene de VP2, que codifica uma das proteínas do capsídeo viral. A prevalência desse vírus foi de 4,8% em crianças menores de cinco anos e de 1% em pacientes maiores de cinco anos. HBoV não foi detectado em crianças sem sintomas. Dos 259 pacientes analisados em 2005, 25 (10%) foram positivos para HBoV. Esse vírus circulou mais frequentemente em abril, mês de maior incidência do HRSV. Em 2006, HBoV foi detectado em apenas 10 LFNs de 334 (3%) amostras testadas, sem qualquer pico de freqüência. Em 2007 HBoV foi detectado em 13 de 552 (2%) amostras, com uma freqüência de detecção um pouco maior em junho e julho. Os sintomas mais comumente observados foram rinorréia, tosse, febre e chiado, que foram observados geralmente em mais de 50% dos casos positivos para HBoV. Não houve uma diferença significativa na prevalência desses sintomas entre as crianças positivas e negativas para HBoV. Entretanto, foi observada uma maior freqüência de diarréia entre as crianças com esse vírus. Nesse estudo também foi documentado uma alta freqüência de co-infecções virais entre os pacientes com HBoV. Os vírus mais frequentemente associados com o bocavirus humano foram: HRSV, HRV e HAdV. Além disso, foi detectado uma maior carga viral media e uma maior freqüência de diarréia nos 15 pacientes com infecção exclusiva por HBoV do que nos pacientes com co-infecção. Esses resultados mostraram que HBoV pode alcançar títulos enormes (tão grandes como1014/mL) em LFNs de pacientes com sintomas respiratórios e que isso é associado a de diarréia. O seqüenciamento do genoma inteiro de HBoV realizado nesse estudo indica que a divergência genômica entre as amostras desse vírus é muito pequena. Como conclusão, nós demonstramos que HBoV circula e é detectado em associação com sintomas de infecção respiratória e diarréia no Brasil. Novos estudos, com um longo acompanhamento em diferentes populações serão necessários para determinar a sazonalidade e o real impacto clínico de HBoV em nosso país. / Human bocavirus (HBoV) is a parvovirus recently identified in association with respiratory tract infections. HBoV 5217 nt genome contains 3 open reading frames encoding four proteins (NS1, NP-1, VP-1 and VP-2). HBoV has been reported in respiratory samples from children in several parts of the world (including Australia, North America, Europe, Asia, and Africa), suggesting that the virus circulates worldwide. However, no longitudinal studies of HBoV in respiratory samples have been reported in Latin America. We report a prospective study of HBoV in nasopharyngeal aspirates (NPAs) collected from patients seen for acute respiratory tract infections (ARI) at the University of Sao Paulo Hospital in Ribeirao Preto, southeast Brazil and at the University Hospital in Salvador, Brazil. 1288 NPAs from 1217 patients was submitted to the virology lab for respiratory virus detection from 2005 to 2007 and were screened for HBoV by polymerase chain reaction (PCR), whom 962 were under 5 years of age and 177 were older than 5 years. In addition, NPAs from 50 children under 12 years without IRA was also tested to HBoV for PCR. All samples positive of HBoV was tested for others respiratory virus, including the human respiratory syncitial virus (HRSV), human rhinovirus (HRV), human influenza (HFLU), human metapneumovirus (HMPV), human parainfluenza virus (HPIV), human coronavirus (HCoV) and human adenovirus (HAdV). These samples had their HBoV viral load determined by real time PCR and the viral entire genome of nineteen HBoV sample was sequenced. We also cloned and expressed in S. cerevisae (Y258) the gene of VP2, one protein of viral capside. The prevalence of this virus was of 4,8% in children under 5 years and 1% in adults, both with IRA. HBoV was not found on the patients without symptoms. In 2005, of the 259 patients tested, 25 (10%) were positive for HBoV. Interestingly, the virus circulated more frequently in April, the month of peak activity of respiratory HRSV. In 2006 HBoV was detected in only 10 NPAs out of 334 samples (3%) tested, without any notable peak of frequency. In 2007 HBoV was detected in 13 out of 552 (2%) tested samples with little higher frequency of detection in June an July. Rhinorrhea, cough, and wheezing were observed in more than 50% of the HBoV-positive children, and no obvious respiratory clinical differences were noted between HBoV-positive and negative children. However, was noted a higher frequency of diarrhea on HBoV-positive patients. In this study was also observed a larger frequency (71%) of viral coinfections between the HBoV-positive patients. The respiratory viruses more frequently associated with human bocavirus were: HRSV, HRV and HAdV. Interestingly, on the 15 HBoV-alone patients was observed a higher viral load and a higher prevalence of diarrhea than HBoV-coinfection patients. These results showed that this virus can reach enormous titles (like 1014) in NPAs from patients with respiratory infection symptoms and this is associated with diahhrea. The entire genome sequencing of HBoV of our study indicates that the genetic divergence between the HBoV lineages is small. In conclusion, we demonstrated that HBoV circulates and is detected in association with respiratory symptoms and diarrhea in Brazil. Long term surveillance will be needed to determine whether or not an HBoV season occurs and what is the real clinical impact of this virus in our country.

O vírus respiratório sincicial (RSV) é responsável por importante morbidade em receptores de transplante de células tronco-hematopoéticas (TCTH), especialmente no período que antecede a enxertia. A imunidade induzida pela infecção pelo RSV é transitória e as reinfecções são freqüentes. O comportamento e papel dos anticorpos anti-RSV em receptores de TCTH é desconhecido. Em amostras de soro estocadas, ensaio imunoenzimático (ELISA) foi aplicado para detecção de anticorpos anti-RSV para avaliar a dinâmica desses anticorpos antes e após o TCTH, em pacientes com e sem infecção pelo RSV, bem como a resposta de anticorpos específicos nos pacientes com infecção pelo RSV diagnosticada por imunofluorescência direta. A mediana do tempo de coleta das amostras pré-TCTH foi de -35 e -44 dias nos casos e controles, respectivamente, com média de títulos de anticorpos anti-RSV de 2490 UA/mL e 2872 UA/mL, respectivamente. Após o transplante, as medianas de tempo das 3 amostras analisadas dos pacientes com infecção pelo RSV foram d+14, d+52 e d+89 e os respectivos títulos de anticorpos foram 2457 UA/mL, 2715 UA/mL e 2950 UA/mL. Nos pacientes sem infecção pelo RSV (controles), as medianas de tempo das 3 amostras analisadas foram d+9, d+69 e d+93 e os respectivos títulos de anticorpos foram 2738 UA/mL, 2794 UA/mL e 2642 UA/mL. Não houve diferença estatística entre os dois grupos. Nenhum dos pacientes com infecção pelo RSV apresentou elevação de quatro vezes nos títulos de anticorpos / Respiratory syncytial virus (RSV) infection can cause significant morbidity and mortality in haematopoietic stem cell transplant (HSCT) recipients, especially when upper respiratory tract infection (RTI) progresses to lower RTI, which is expected to occur in more than 50% of the patients. The humoral immunity induced by RSV infection is transient and reinfections are frequent. The dynamics and role of anti-RSV antibodies in HSCT recipients are unknown. In stored serum samples, an enzyme-linked immunosorbent assay (EIA) was applied to evaluate the dynamics of anti-RSV antibodies in HSCT recipients with and without RSV infection, as well as the specific humoral response in HSCT recipients with RSV infection diagnosed by direct immunofluorescent assay in nasal wash samples. Pre-transplant samples were selected at a median time of 35 and 44 days and the mean concentration of RSV antibodies were 2490 AU/mL and 2872 AU/mL, in cases and controls, respectively. After HSCT, serum samples from patients with RSV infection (cases) were evaluated at median time of 14, 52 and 89 days, and the respective mean concentrations of anti- RSV antibodies were 2457 AU/mL, 2715 AU/mL and 2950 AU/mL. In patients without RSV (controls), serum samples were evaluated at median time of 9, 69 and 93 days, and the respective mean concentrations of anti-RSV antibodies were 2738 AU/mL, 2794 AU/mL e 2642 AU/mL. Difference was not statistically significant. No patient developed a four-fold rise in RSV antibody titers

S.aureus resistente à meticilina(SARM) é um problema em centros de fibrose cística(FC). Foi desenvolvido um multiplex PCR(mPCR) para detecção do SARM em secreção respiratória de 106 pacientes com FC. Foram usados 3 pares de primers para amplificar os genes: mecA, coa, 16S rRNA. O mPCR detectou até 0,25pg de DNA de SARM e identificou 70/106(66,0%) pacientes com S.aureus e 28/106(26,4%) com SARM. O mPCR mostrou especificidade, sensibilidade, valores preditivos positivo e negativo de 87,8%, 84,4%, 50% e 97,5%, considerando a cultura como padrão-ouro. Os resultados discordantes foram testados com outros primers, confirmando os obtidos pelo mPCR em 82/84. O mPCR mostrou-se método rápido e confiável para detecção de SARM / Methicillin-resistant S.aureus(MRSA) is a significant concern in cystic fibrosis(CF) centers. A multiplex PCR(MPCR) was developed to detect MRSA in respiratory secretion of 106 CF patients. Three pairs of primers were used for amplification of genes: mecA, coa, 16S rRNA. MPCR detected 0.25pg of MRSA DNA and identified 70/106(66.0%) of patients with S.aureus and 28/106(26.4%) with MRSA. MPCR showed specificity, sensitivity, positive and negative predicted values at 87.8%, 84.4%, 50% and 97.5%, considering culture as the gold standard. Discrepant results were retested using different primers, and confirmed MPCR results in 82/84. The developed MPCR was found to be a rapid and reliable method for MRSA detection