Made available in DSpace on 2016-01-07T13:36:23Z (GMT). No. of bitstreams: 2 andressa_martorelli_ioc_mest_2014.pdf: 1406086 bytes, checksum: aa1b67f7a16391a611cf88b0ce6ca128 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2015-11-23 / Indivíduos infectados pelo HIV têm um risco mais elevado de serem afetados por doenças graves, tais como infecções por vírus respiratórios, incluindo o vírus da influenza. A imunossupressão desses indivíduos pode afetar a sua capacidade de resposta à imunização ativa. A vacinação contra o vírus influenza ainda representa a principal forma de reduzir o impacto deste vírus. Devido à circulação em 2009 do vírus influenza pandêmico A (H1N1) pdm09 e influenza sazonal A (H3N2) e vírus B, a composição atual vacina incluem antígenos destes três agentes em sua formulação. Assim, a análise do impacto da vacina trivalente inativada contra influenza (TIV) em indivíduos infectados pelo HIV merece mais estudos. Uma coorte de 131 indivíduos HIV- 1 positivos, com viremia controlada recebeu duas doses únicas de TIV com 21 dias de intervalo. Os títulos de anti- hemaglutinantes (HA) de seus soros foi avaliada em diferentes pontos de tempo (dias zero, 21 e 42, bem como seis meses pós- vacinação). No que diz respeito à resposta imune ao vírus A(H1N1) pdm09 , observouse que um ano depois de ter recebido a vacina monovalente contra este antígeno , 62,6% dos indivíduos permaneceram soroprotegidos, uma queda de 20% em relação à 6 meses pósvacinação monovalente no ano de 2010. Além disso, após a primeira dose de TIV, títulos soroprotetores foram encontrados em todos os indivíduos A soroproteção de baseline para os antígenos H3N2 e influenza B foi de 67,9% e 27,5%, respectivamente. Em relação à soroconversão, observamos que 19,8% e 21% dos indivíduos apresentavam títulos antihemaglutinantes para o vírus A(H1N1)pdm09, 21 e 42 dias pós-vacinação com TIV, respectivamente. A soroconversão para o antígeno H3N2 foi 15,7% e 16% aos 21 e 42 dias respectivamente. Em relação à Influenza B, 35,7% e 38,6% dos indivíduos soroconverteram aos 21 e 42 dias após a vacinação. Aos seis meses pós-vacinação, os títulos de anti \2013 hemaglutinantes dos soros caiu para abaixo 15,7% para o antígeno A(H1N1)pdm09, 14,4% para H3N2 e 29,1% para antígenos de influenza B. Observamos também que indvíduos com maiores contagem de células CD4 resposram melhor a vacinação frente ao virus influenza B. Em conjunto, nossos resultados indicam que indíviduos HIV-1 positivos apresentam melhores repostas humorais para os antígenos A(H1N1)pdm09, quando comparado com os outros dois antígenos presentes na TIV / HIV - infected individuals have a higher risk of being affected by serious illnesses , such as in fections by respiratory viruses , including influenza virus . Immunosuppression of these individuals can affect their ability to respond to active immunization . Vaccination against influenza is still the main way to reduce the impact of this virus . Due to the movement in 2009 pandemic influenza A (H1N1) pdm09 and seasonal influenza A (H3N2) and B viruses , the current vaccine composition include antigens of these three agents in their formula tion . Thus, analysis of the impact of trivalent inactivated influenza vaccine (TIV ) in HIV - infected individuals deserves further study. A cohort of 131 HIV - 1 positive individuals with controlled viremia received two single dose s of TIV 21 days apart. Th e titers of anti - hemagglutinant (HA ) of their sera was assessed at different time points (zero, 21, 42 days a nd six months post - vaccination) . With re gard to immune to the virus A (H1N1) pdm09 response , it was noted that a year af ter having received the monovalent vaccine against this antigen , 62.6 % of subjects remained seroprotection , a decrease of 20 % compared to 6 monovalent months post - vaccination in 2010 . Moreover, after the first dose of TIV , seroprotective titers were f ound in all individuals . The seroprotection baseline for H3N2 and influenza B antigens was 67.9 % and 27.5 % , respectively . Regarding to seroconversion , we found that 19.8 % and 21 % of subjects had anti - hemag glutinin titles for the A (H1N1)pdm09 , 21 and 42 days post - vaccination with TIV , respectively. Seroconversion for H3N2 antigen was 15.7% and 16 % at 21 and 42 days resp ectively. Regarding Influenza B , 35.7 % and 38.6 % of subjects seroconverted after 21 and 42 days after vaccination. At six months po st - vac cination , the titers of anti - hemagglutinin sera fe ll below 15.7 % for antigen A (H1N1)pdm09 , 14.4% to 29.1 % for H3N2 and influenza B antigens . We also observed that indvíduos with higher CD4 cell counts resposram best vaccination against the influ enza virus B. Together, our results indicate that HIV - 1 positive individuals show better humoral responses to antigens A (H1N1) pdm09 compared with others two antigens present in the TIV. / HIV - infected individuals have a higher risk of being affected by serious illnesses , such as in fections by respiratory viruses , including influenza virus . Immunosuppression of these individuals can affect their ability to respond to active immunization . Vaccination against influenza is still the main way to reduce the impact of this virus . Due to the movement in 2009 pandemic influenza A (H1N1) pdm09 and seasonal influenza A (H3N2) and B viruses , the current vaccine composition include antigens of these three agents in their formula tion . Thus, analysis of the impact of trivalent inactivated influenza vaccine (TIV ) in HIV - infected individuals deserves further study. A cohort of 131 HIV - 1 positive individuals with controlled viremia received two single dose s of TIV 21 days apart. Th e titers of anti - hemagglutinant (HA ) of their sera was assessed at different time points (zero, 21, 42 days a nd six months post - vaccination) . With re gard to immune to the virus A (H1N1) pdm09 response , it was noted that a year af ter having received the monovalent vaccine against this antigen , 62.6 % of subjects remained seroprotection , a decrease of 20 % compared to 6 monovalent months post - vaccination in 2010 . Moreover, after the first dose of TIV , seroprotective titers were f ound in all individuals . The seroprotection baseline for H3N2 and influenza B antigens was 67.9 % and 27.5 % , respectively . Regarding to seroconversion , we found that 19.8 % and 21 % of subjects had anti - hemag glutinin titles for the A (H1N1)pdm09 , 21 and 42 days post - vaccination with TIV , respectively. Seroconversion for H3N2 antigen was 15.7% and 16 % at 21 and 42 days resp ectively. Regarding Influenza B , 35.7 % and 38.6 % of subjects seroconverted after 21 and 42 days after vaccination. At six months po st - vac cination , the titers of anti - hemagglutinin sera fe ll below 15.7 % for antigen A (H1N1)pdm09 , 14.4% to 29.1 % for H3N2 and influenza B antigens . We also observed that indvíduos with higher CD4 cell counts resposram best vaccination against the influ enza virus B. Together, our results indicate that HIV - 1 positive individuals show better humoral responses to antigens A (H1N1) pdm09 compared with others two antigens present in the TIV

RIBEIRO, Joyce Fonteles. Perfil clínico-epidemiológico das infecções respiratórias agudas causadas por metapneumovírus humano em crianças atendidas no Hospital Infantil Albert Sabin - Fortaleza/Ceará. 2008. 94 f. Dissertação (Mestrado em Microbiologia Médica) - Universidade Federal do Ceará. Faculdade de Medicina, Fortaleza, 2008. / Submitted by denise santos (denise.santos@ufc.br) on 2012-01-05T12:12:30Z No. of bitstreams: 1 2008_dis_jfribeiro.pdf: 2268899 bytes, checksum: 31df8e028e25d45794437cdab4883903 (MD5) / Approved for entry into archive by Eliene Nascimento(elienegvn@hotmail.com) on 2012-02-02T16:25:35Z (GMT) No. of bitstreams: 1 2008_dis_jfribeiro.pdf: 2268899 bytes, checksum: 31df8e028e25d45794437cdab4883903 (MD5) / Made available in DSpace on 2012-02-02T16:25:35Z (GMT). No. of bitstreams: 1 2008_dis_jfribeiro.pdf: 2268899 bytes, checksum: 31df8e028e25d45794437cdab4883903 (MD5) Previous issue date: 2008 / The human metapneumovírus (hMPV) is a newly discovered virus that has been considered as one of the most common agents of acute respiratory infections (ARI) virus in childhood. The objectives of this study were: 1) to observe the frequency of infections caused by hMPV among children attending Hospital Infantil Albert Sabin, a major pediatric hospital in Ceará, from January 2006 to December 2007; 2) to describe aspects of seasonality of these infections relating them to the occurrence of rain and the circulation of other respiratory viruses, 3) to describe the clinical and epidemiological characteristics of patients infected by hMPV, compared with positive and negative patients for other viruses; 4) to evaluate the IFI assay as a method of diagnosis for the detection of hMPV. Nasopharyngeal aspirates were collected from children with symptoms of ARI and submitted to indirect immunofluorescence assays for the detection of the following respiratory viruses: hMPV, respiratory syncytial virus (RSV), influenza A and B, adenovirus and parainfluenza 1, 2 and 3. During the 24 months of study, samples were collected from 1276 and respiratory viruses were demonstrated in 380 (29.78%) samples. The hMPV was the second most frequently detected respiratory viruses representing a total of 8.69% of all samples and 29% among the samples positive for the virus analyzed. It was not observed for hMPV a pattern of seasonality or correlation with the rainy season. Most patients positive for hMPV were attended in the emergence (89.2%). The mean age of patients infected by hMPV was 27 months, wich is significantly older than that for VSR (15 months), adenovirus (14 months) and parainfluenza virus 3 (18 months). Among patients infected by hMPV, 53.2% had a diagnosis of infections of the upper airways and 46.7% had a diagnosis of infections of the lower airways. The hMPV infections showed the same spectrum of infections caused by other viruses analyzed. The hMPV associated to more cases of pneumonia that led to the hospitalization of children infected than other viruses analyzed. More than half of these patients used the aerosol / salbutamol as conduct therapy (68.5%). The IFI assay proved to be quite effective as a method of diagnosis for the detection of hMPV in this study / O metapneumovírus humano (MPVh) é um vírus que tem se destacado como um dos agentes mais freqüentes de infecções respiratórias agudas (IRA) virais na infância. Este estudo teve como objetivos: observar a freqüência das infecções causadas pelo MPVh em crianças atendidas por IRA no Hospital Infantil Albert Sabin, hospital pediátrico de referência do estado do Ceará, no período de janeiro de 2006 a dezembro de 2007; descrever aspectos de sazonalidade dessas infecções relacionando-as com a ocorrência de chuvas e a circulação de outros vírus respiratórios; descrever as características clínico-epidemiológicas dos pacientes infectados pelo MPVh, comparando com os pacientes negativos e com os positivos para outros vírus e avaliar a técnica de IFI como método de diagnóstico para a detecção do MPVh. Amostras de secreção de nasofaringe foram coletadas de crianças com sintomas de IRA e submetidas à técnica de imunofluorescência indireta para detecção dos seguintes vírus respiratórios: MPVh, vírus sincicial respiratório (VSR), influenza A e B, adenovírus e parainfluenza 1, 2 e 3. Durante os 24 meses de estudo, foram colhidas amostras de 1276 crianças sendo detectado algum vírus respiratório em 380 (29,78%) amostras. O MPVh foi o segundo vírus respiratório mais encontrado representando um total de 8,69% de todas as amostras e de 29% dentre as amostras positivas para os vírus pesquisados. Não foi observado para o MPVh um padrão de sazonalidade nem correlação com período chuvoso. A maioria dos pacientes positivos para MPVh foram atendidos na emergência (89,2%). A média de idade dos pacientes positivos para o MPVh foi de 27 meses sendo significativamente superior que a das crianças infectadas pelo VSR (15 meses), adenovírus (14 meses) e vírus parainfluenza 3 (18 meses). Dentre os pacientes infectados pelo MPVh, 53,2% tiveram o diagnóstico de infecções das vias aéreas superiores e 46,7% tiveram o diagnóstico de infecções das vias aéreas inferiores. As infecções por MPVh apresentaram o mesmo espectro de infecções causadas pelos demais vírus pesquisados. O MPVh associou-se mais a casos de pneumonia que levaram à hospitalização das crianças infectadas do que outros vírus analisados. Mais da metade dos pacientes infectados pelo MPVh utilizaram o aerossol / salbutamol no seu tratamento (68,5%). A técnica de IFI mostrou-se bastante eficaz como método de diagnóstico para a detecção do MPVh nesse estudo

O vírus influenza A (IAV) é um agente zoonótico de grande relevância tanto para saúde humana como animal. A influenza suína teve seu primeiro reconhecimento clínico em 1918, em suínos do Meio Oeste dos EUA, coincidindo com a pandemia de influenza em humanos. Desde então, o IAV permanece como um importante patógeno para a indústria suinícola em todo o mundo. A grande variabilidade genética destes vírus é causada por dois principais mecanismos genéticos: mutações pontuais e recombinações genéticas. A influenza é endêmica em muitos países e a emergências de recombinantes tem desafiado o controle e o diagnóstico desta enfermidade. No Brasil, a infecção pelo IAV em suínos (swIAV) não está bem caracterizada; poucos relatos evidenciam a prevalência deste agente antes do ano de 2009, especialmente no Estado do Rio Grande do Sul, que alberga um dos maiores rebanhos de suínos do Brasil. Em vista disso, este trabalho teve como objetivo investigar ocorrência de swIAV em alguns rebanhos suínos comerciais do Estado do Rio Grande do Sul, Brasil, no período de 2013-2014, e determinar os tipos e subtipos de vírus circulantes naquelas propriedades. O primeiro capítulo deste estudo reporta os aspectos clínicos, patológicos e virológicos da ocorrência de influenza suína e co-infecções identificadas em seis propriedades suinícolas selecionadas na região do Vale do Taquari. Neste estudo foram analisados suabes nasais coletados de 66 animais e 6 amostras de tecido pulmonar de suínos com sinais de infecção respiratória. A detecção viral foi feita através de uma PCR de triagem e confirmada através do isolamento viral em células MDCK. A identificação dos subtipos virais foi feita através de uma PCR em Tempo Real (rRT-PCR) para o subtipo A(H1N1)pdm09 ou através de uma PCR multiplex (RT-PCR) para outros subtipos de swIAV. A detecção de agentes bacterianos foi realizada apenas nas amostras de tecido pulmonar, através da pesquisa de genomas bacterianos por PCR. O subtipo A(H1N1)pdm09 foi identificado em 4/6 granjas e o subtipo H1N2 em 2/6 granjas. Além disso, agentes envolvidos no complexo respiratório dos suínos foram identificados em todas as granjas; Pasteurella multocida foi identificada em 5/6 granjas e Mycoplasma hyopneumoniae em 3/6 granjas. Actinobacillus pleuropneumoniae (1/6), Haemophilus parasuis (1/6) e PCV2 (1/6) também foram detectados. O segundo capítulo deste estudo teve como objetivo o sequenciamento do genoma completo de um novo recombinante H1N2 de origem humana, detectado em suínos. O genoma completo foi gerado através de uma RT-PCR. Os produtos foram purificados e submetidos ao sequenciamento utilizando a plataforma MiSeq (illumina). A análise filogenética revelou que as sequencias dos genes HA e NA correspondem a genes de IAV de origem humana, enquanto que as sequencias dos genes que codificam as proteínas internas do vírus (PB1, PB2, PA, NP, M e NS) correspondem a genes de amostras do vírus A(H1N1)pdm09. O terceiro capítulo reporta o sequenciamento completo dos genomas de 8 amostras de vírus influenza identificados nas populações de suínos amostradas. Foram identificados dois subtipos virais de origem humana (H1N2 e H3N2), além do vírus A(H1N1)pdm09. Os subtipos de origem humana possuem os genes HA e NA similares a vírus sazonais de humanos e os genes internos são estreitamente relacionados com o vírus A(H1N1)pdm09. / Influenza A virus (IAV) is a zoonotic agent of great relevance to human and animal health. Swine influenza was first recognized clinically in pigs in the Midwestern U.S., in 1918, coinciding with the human influenza pandemic. Since that time swine influenza has remained of importance to the swine industry throughout the world. The great genetic variability of influenza viruses is caused by two main genetic mechanisms: point mutations (antigenic drift) and gene reassortment (antigenic shift). Influenza is endemic in pigs in many countries and the emergence of new viruses has been challenging its control and diagnostics. Influenza virus (swIAV) infection in Brazilian swine population is not well characterized, and little evidence existed of swIAV circulation before 2009, especially in Rio Grande do Sul State, which hosts one of the largest swine populations in Brazil. Thus, this study aimed to investigate the occurrence of IAV in commercial swine herds in the state of Rio Grande do Sul, Brazil, between 2013-2014 and to know the types and subtypes of swine influenza viruses that are circulating in these herd. The first chapter of this study reports the clinical, pathological and virological aspects of the occurrence of swine influenza and related co-infections in six pig properties of the Taquari Valley region. In this study were analyzed nasal swabs collected from 66 animals and six lung tissue samples from pigs showing clinical signs of respiratory disease. IAV detection was performed by PCR screening and confirmed by virus isolation in MDCK cells and hemagglutination (HA). Influenza A subtyping was performed by real-time reverse transcription PCR (rRT-PCR) to detect the 2009 H1N1pandemic A(H1N1)pdm09; other swIAV subtypes were identifieded by multiplex RT-PCR. Bacterial infections were identified through detection of bacterial genomes by PCR, only in lung samples. Influenza A was detected by screening PCR in 46/66 swab samples and from 5/6 lungs. Virus was recovered from pigs of the six herds. Subtype A(H1N1)pdm09 was detected in 4/6 herds and H1N2 in the other 2/6 herds. In lung tissues, further agents involved in porcine respiratory disease complex were detected in all cases; Pasteurella multocida was identified in 5/6 samples and Mycoplasma hyopneumoniae in 3/6. Actinobacillus pleuropneumoniae (1/6), Haemophilus parasuis (1/6) and PCV2 (1/6) were also detected. The aim of the second chapter was to sequence the whole-genome of a novel human-like H1N2 swine influenza virus. Wholegenome sequences were generated by RT-PCR. Amplicons were purified followed by sequencing in the MiSeq sequencing platform (Illumina). Phylogenetic analyses revealed that the HA and NA genes clustered with influenza viruses of human lineage, whereas the internal genes (PB1, PB2, PA, NP, M and NS) clustered with the A(H1N1)pdm09. The third chapter reports the genetic sequencing of the full genomes of eight swine influenza viruses circulating in the sampled pig population. Two swine human-like subtypes (H1N2 and H3N2) and the A(H1N1)pdm09 virus were identified. The human-like subtypes have the HA and NA genes similar to the human seasonal strains and the internal genes are closely related to the virus A(H1N1)pdm09.

Humans are subject to infection with a wide range of commensal and pathogenic organisms. Each pathogen requires an appropriate immune response to eliminate or control the invading organism and minimise pathology. Many pathogens have evolved strategies to subvert or manipulate the immune response and establish on-going infections. Similarly acute respiratory infection with virulent strains of influenza A virus are often poorly controlled by the immune system and can cause severe immunopathology and even fatality as a result of an inappropriate and excessive inflammatory response called a ‘cytokine storm’. Morbidity due to influenza infection and exacerbation by the immune response can vary greatly between individuals. The effect of underlying infection on the immune system could contribute to the variation in response. The aim of this project was therefore to determine if co-infection with two pathogens that establish on-going infections could alter the immune response to influenza A and impact the outcome of infection. Persistent infections with filarial helminths can cause debilitating disease and significantly impact the immune response toward a skewed TH2 or regulatory phenotype in order to control pathology. In contrast, infection with gammaherpesviruses in an immunocompetent host causes an initial inflammatory ‘anti-viral’ response before becoming an asymptomatic, latent infection. In an immunocompromised host, gammaherpesviruses can reactivate and lead to clinical presentation of disease. This suggests that these viruses require an on-going immune response to control all stages of infection. Both filarial helminths and gammaherpesviruses are common infections in human populations and therefore mouse models of these infections provide relevant systems to study their potential role in influenza virus infections. In a BALB/c murine co-infection model, latent infection with the rodent gammaherpesvirus MHV-68 led to significantly decreased weight loss and clinical signs following high dose infection with A/WSN/33, (a H1N1 influenza A virus). This was coupled with decreased immunopathology in the lung and fewer infiltrating lymphocytes in the alveolar spaces and around larger airways, although infectious virus titres were not significantly reduced. This response was coupled with a decreased production of inflammatory cytokines and chemokines in co-infected mice 6 days post infection which correlated with the amelioration of pathogenesis in these animals. A repeat of the study in 129Sv/Ev IFNγR knock out mice showed the same protective effect in the co-infected mice, suggesting IFNγ is not critical for the protective phenotype. Mice infected with latent MHV-68 alone showed a significant increase in expression of T cell chemokines in the lung and alveolar macrophages had a significantly increased production of suppressor of cytokine signalling (SOCS-1) suggesting latent MHV-68 infection may impact the phenotype of macrophages in the lung, modulating the response to influenza co-infection. A co-infection model with a persistent rodent filarial helminth, Litomosoides sigmodontis and A/WSN/33 was also established in BALB/c mice. The L4 developmental stage of L. sigmodontis infection had no impact on co-infection with A/WSN/33. Adult stage worms, however, appeared to have a protective effect against A/WSN/33 pathogenesis. Co-infected mice had significantly delayed weight loss and clinical signs 3-5 days post infection. CD4+ and CD8+ T cells in the lung draining lymph nodes had significantly reduced TH1 and TH2 phenotypes (measured by cytokine production) compared with singly infected controls. IFNγ secreting CD4+ T cells in the lungs of co-infected mice also secreted increased levels of IL-10, suggesting an increase in regulation of the inflammatory response to A/WSN/33. At the full patent stage of L. sigmodontis infection, co-infection with A/WSN/33 led to increased clinical signs and significantly exacerbated weight loss. CD4+ and CD8+ T cells in the lung draining lymph nodes were inflammatory in L. sigmodontis infected mice alone as well as co-infected mice and there were no differences in the percentage of CD4+ T cells in the lung secreting IL-10 and IFNγ between co-infected and influenza infected mice. A loss in regulatory responses during the patent stage of L. sigmodontis infection may therefore contribute to the loss of protection against A/WSN/33 at this time point within the co-infection model. Understanding the impact of an underlying infection on the immune system could provide immune mechanisms that could be exploited to increase vaccine efficacy against influenza and similarly help to provide better treatment for individuals infected with influenza A. These results may also help predict the outcome of influenza A infection in individuals already infected with highly immunogenic, on-going infections.

Trata-se de um estudo sobre o impacto da introdução da vacina contra influenza no perfil de morbimortalidade por pneumonias na população acima de 65 anos, em Porto Alegre, no período de 1995 a 2001. Associado ao estudo da série histórica, investigamos os motivos de adesão ou recusa à prática de vacinação contra influenza entre 138 idosos. Aprofundamos a análise das concepções do processo saúde-doença e das práticas preventivas em saúde com um grupo de 30 idosos usuários do sistema público de saúde. A metodologia utilizada é de caráter epidemiológico do tipo série temporal, combinando entrevistas com idosos e categorização temática das informações obtidas. Os dados de morbidade foram obtidos pela pesquisa documental estatística, a partir dos dados de internação hospitalar do Sistema Único de Saúde (Tabwin) e os dados de mortalidade foram obtidos a partir do Sistema de Informação em Mortalidade (SIM). Os resultados apontam um comportamento de tendência à queda na morbidade, através das internações hospitalares e na mortalidade por pneumonias, após a introdução regular da vacina contra influenza em nosso meio no ano de 1999. Particularizando a adesão ou recusa à prática de vacinação, os idosos demonstram que fatores culturais e sociais influenciaram suas decisões. Os idosos que participaram deste estudo também revelaram que é fundamental a manutenção de uma atividade física, intelectual ou laboral para um envelhecimento saudável. Os resultados deste estudo, acredita-se, contribuem para o aperfeiçoamento das práticas de promoção em saúde através da educação em saúde e da adoção de medidas de proteção específica como a vacinação com eficácia e credibilidade junto à população.

Influenza virus H1N1 is a huge threat on human health. Influenza occurs with seasonal variations and reaches peak prevalence in winter, with many people killed worldwide every year. In the research of interaction between influenza virus and cells, four major parts were in the range of our consideration, namely the proteins of virus, the proteome of host cell, the method of proteomic and the potencial medicine related with those significant proteins. Hemagglutinin (HA), as an envelope protein, plays an important role in influenza A virus. It was found that HA has a series of isoforms in two dimensional gels in this study. For the investigation of HA, firstly, virus was purified by sucrose density-gradient centrifugation, followed by the separation of virus proteins through electrophoresis method, and then these proteins were digested by different enzymes and analyzed through MALDI-TOF MS and ESI-Q-TOF MS. Database searching was used for identification of sequences. The results of the virus samples digested by different enzymes were compared, and the isoforms of HA were proved to be related with the glycan and their glycosylation sites. A novel strategy of stable-isotope N-phosphorylation labeling was developed for peptide de novo sequencing and protein quantification based on organic phosphorus chemistry. Different from other stable-isotope labeling reagents that needed to be activated in advance for peptide coupling, N-phosphorylation labeling reagents were activated in situ to form labeling intermediates with high activity and selectivity targeting on N-terminus and -amino group of lysine under various reaction conditions. The obtained results showed excellent correlation of the measured ratios to theoretical ratios with errors that ranging from 0.5 to 6.7 % and relative standard deviation of less than 10.6 %, indicating the reproducibility and preciseness of the developed method. The method development based on organic phosphorus chemistry offered a new approach for quantitative proteomics by using novel stable-isotope labeling reagents. A method combining hydrazide chemistry, stable isotope labeling and mass spectrometry analysis was developed and applied to study glycoproteins of H1N1 (A/Purto Rico/8/1934) infected cell line (A549). The result showed that some glycoproteins were significant in influenza virus infected cells. In these glycoproteins, RPC1_HUMAN, RHG25_HUMAN , RPTOR_HUMAN, ARHGC_HUMAN, ROCK1_HUMAN, DOCK3_HUMAN were down-regulated. Protein named TITIN_HUMAN, DESP_HUMAN, PTN13_HUMAN were up-regulated. High dose of N-acetylcysteine (NAC) was recently reported for a therapy of H1N1 influenza pneumonia. NAC was used as a small-molecule organic probe to investigate the protein expression of human lung carcinoma cell line (A549) infected by influenza virus H1N1. The obtained results showed that NAC kept cells away from apoptosis. Virus-infected cells were arrested in G0/G1 phase. The lowest cell population of G0/G1 phase was detected when the cells were treated by 10 mM NAC for one day. Software analysis showed that 4 proteins had close relationship. The results indicated that NAC as a small-molecule probe might effect the proteins expression of A549 cells infected by the H1N1 virus

No description available.

Aim: To temporally describe the hospital morbidity in the elderly population due to influenza related causes and its complications and to estimate the effects of the vaccine over its coefficients in Brazilian Southern State of Santa Catarina. Methods: An ecological study combining temporal series from 1995 to 2009 was carried out. Secondary data from the Brazilian Sistema de Informações Hospitalares was used. The study's population was composed by records of 60 year old individuals and older, who was living in the nine macroregions of health of the state. To observe the number of hospital admissions, influenza related causes and its complications according to the International Classification of Diseases were used. It was estimated the coefficient of hospital admissions according to gender and age for the state, the death rate regardless the sex and age, the seasonal effect. The effect of seasonality and vaccine were measured using multiple linear regressions. Results: Males and the group of age of 80 and over showed higher rates of hospital admissions. Variations were observed, with a decrease of 15.8% in the first quarters of the years of the study, and an increase of 14.9% in the thirds quarters highlighting the effects of seasonality. It was also observed a decrease of 18.9% from 1999 on. Conclusion: It was observed important effects on the hospitalization admissions rates in Santa Catarina associated to seasonality and to the introduction of the influenza vaccination campaigns in the studied period. / Submitted by Rogele Pinheiro (rogele.pinheiro@unisul.br) on 2018-02-28T16:59:18Z No. of bitstreams: 1 104838_Alana.pdf: 2108859 bytes, checksum: ff0050caead0675d80f419d9dd249bd5 (MD5) / Approved for entry into archive by Ana Barreiros (ana.barreiros@unisul.br) on 2018-02-28T17:01:02Z (GMT) No. of bitstreams: 1 104838_Alana.pdf: 2108859 bytes, checksum: ff0050caead0675d80f419d9dd249bd5 (MD5) / Made available in DSpace on 2018-02-28T17:01:02Z (GMT). No. of bitstreams: 1 104838_Alana.pdf: 2108859 bytes, checksum: ff0050caead0675d80f419d9dd249bd5 (MD5) Previous issue date: 2011 / Objetivos: Descrever temporalmente a morbidade hospitalar na população idosa por causas relacionadas à influenza e suas complicações e estimar os reflexos da vacina sobre seus coeficientes no Estado de Santa Catarina. Métodos: Estudo epidemiológico de delineamento ecológico combinando séries temporais de 1995 a 2009. Utilizaram-se dados secundários oriundos do Sistema de Informações Hospitalares do Sistema Único de Saúde do Brasil. A população do estudo foi composta por registros de indivíduos de 60 anos e mais, residentes nas nove macrorregiões de saúde do Estado de Santa Catarina. Para o cálculo dos coeficientes de internação hospitalar foram utilizadas causas relacionadas à influenza e suas complicações de acordo com a Classificação Internacional de Doenças. Foram analisados os coeficientes de internação hospitalar por sexo e faixa etária para o Estado, a tendência da morbidade sem distinção de sexo e faixa etária, o efeito da sazonalidade. Utilizando a análise regressão linear múltipla calculou-se o efeito exercido pela sazonalidade e os reflexos causados com a introdução da vacina contra influenza no Estado de Santa Catarina. Resultados: O sexo masculino e a faixa etária acima de 80 anos apresentaram coeficientes mais elevados. Observaram-se variações, com diminuição de 15,8% nos primeiros trimestres dos anos de estudo, e aumento nos terceiros trimestres em 14,9%, evidenciando o reflexo da sazonalidade. O efeito da vacinação, a partir de 1999, foi de 18,4%, com relação ao efeito que exerce na diminuição do coeficiente de internação. Conclusão: A sazonalidade e a introdução das campanhas de vacinação contra influenza exerceram reflexos nos coeficientes de internação hospitalar no Estado de Santa Catarina no período estudado.

O presente trabalho aborda como questão central o desfecho das gestações expostas ao vírus Influenza A (H1N1) e, consequentemente, ao seu tratamento com o fármaco oseltamivir durante a pandemia do ano 2009. O vírus influenza A H1N1 é produto de vários rearranjos genéticos entre cepas dos vírus influenza previamente circulantes, alguns destes exclusivos de suínos ou de aves, e que se tornaram capazes de infectar humanos. A epidemia de influenza A (H1N1) teve início no México, e expandiu-se rapidamente para países do mundo inteiro, sendo declarada pandemia pela Organização Mundial da Saúde (OMS), aproximadamente dois meses após o aparecimento dos primeiros casos. As gestantes são consideradas um grupo de risco para complicações graves relacionadas ao vírus influenza H1N1, com grande morbidade e mortalidade observadas em epidemias anteriores do vírus Influenza. Quanto aos efeitos sobre o embrião-feto, os estudos a respeito do potencial teratogênico do vírus influenza ainda são limitados. A literatura não demonstrou, até o momento, efeitos adversos desse vírus sobre o embrião-feto. O tratamento específico consiste no uso de inibidores da neuraminidase, zanamivir e oseltamivir, dos quais apenas o último está disponível no Brasil. O objetivo geral da tese é avaliar as gestações expostas ao vírus H1N1 e submetidas ao tratamento com Fosfato de Oseltamivir. Os objetivos específicos são comparar gestantes expostas e não-expostas ao vírus Influenza A H1N1 quanto aos desfechos maternos e perinatais; avaliar os potenciais efeitos adversos da medicação em gestantes expostas ao oseltamivir; e avaliar a saúde e o desenvolvimento neuropsicomotor das crianças expostas durante a gravidez ao oseltamivir. Foi realizado um estudo de coorte prospectivo não controlado que avaliou gestantes com exposição ao vírus H1N1 e ao tratamento com Fosfato de Oseltamivir. A amostra consistiu nas 589 gestantes com sintomas suspeitos de Influenza A notificadas no Sistema de Informação de Agravos de Notificação - Influenza (SINAN-Influenza banco de dados do estado do Rio Grande do Sul). Os seguimentos de 424 gestantes foram realizados por contato telefônico, visita domiciliar, dados de prontuário médico ou Declaração de Nascido Vivo, por uma equipe treinada. . Foram obtidos 243 resultados de exames de PCR (polymerase chain reaction). Houve 163 (67%) casos confirmados de H1N1 e 80 (33%) Influenza não-H1N1. Houve 24 óbitos maternos, sendo 18 em H1N1. Houve 8 natimortos, sendo 5 filhos de gestantes expostas ao H1N1. Não houve diferença nos desfechos perinatais. Apenas um caso de malformação congênita (fenda palatina) foi observado em um bebe não exposto ao oseltamivir. Uso de oseltamivir foi identificado em 221 pacientes. Dessas, 86 gestantes apresentaram PCR positivo para Influenza A (H1N1) e 51 estavam no grupo não- H1N1. Reações adversas foram relatadas em 92 (42%) gestantes. Houve um maior número de reações adversas relatadas em pacientes não-H1N1 após o uso do oseltamivir. Ocorreram menos óbitos maternos (7,2%) nas que receberam oseltamivir comparativamente a 34,7% das mulheres que não foram tratadas (OR: 0,14, IC95%: 0,04-0,42, p=0,0003). Da mesma forma a frequência de natimortos foi menor (2,2%) nas tratadas, em comparação a 13,0% das não tratadas (OR: 0,15, IC95%: 0,03-0,89, p=0,03). Atrasos afetando dois ou mais marcos do desenvolvimento foram relatados em 10 (19,2%) de 52 crianças expostas ao oseltamivir durante o período gestacional e seguidas por no mínimo 36 meses. Essa frequência está acima do esperado para a população brasileira (15%). Em conclusão, espera-se que o presente trabalho seja capaz de contribuir para um melhor entendimento a respeito do potencial teratogênico do vírus Influenza A (H1N1) e de seu tratamento com o fármaco oseltamivir. Estudos futuros serão decisivos no estabelecimento de condutas clínicas no que diz respeito ao tratamento e manejo geral dessa condição nesse grupo específico de pacientes. / The present investigation approaches as central issue the outcomes of pregnancies exposed to the Influenza A (H1N1) virus and, consequently, to its treatment with the drug oseltamivir during the pandemic in the year 2009. The Influenza A H1N1 virus is the product of multiple genetic rearrangements among strains of influenza that had previously been circulating. Some of these were unique to swine and birds and became capable of infecting humans. The influenza A (H1N1) epidemic began in Mexico and rapidly spread to other countries around the world and was declared a pandemic by the World Health Organization (WHO) approximately two months after the first cases appeared. Pregnant women are considered to be a group at risk of serious complications related to the H1N1 influenza virus, with high morbidity and mortality observed in previous Influenza virus epidemics. As for effects on the embryo/fetus, there are still few studies on the teratogenic potential of the influenza virus. The literature has not demonstrated, so far, adverse effects of this virus on the embryo/fetus. The specific treatment is the use of the neuraminidase inhibitors, zanamivir and oseltamivir, of which only the latter is available in Brazil. The general aim of this work is to evaluate pregnancies exposed to the Influenza A (H1N1) virus and submitted to treatment with Oseltamivir Phosphate. Specific objectives are to compare pregnant women exposed and not exposed to the Influenza A H1N1 virus as on maternal and perinatal outcomes; evaluate potential adverse effects of medication in pregnant women exposed to oseltamivir; and evaluate the health and neurodevelopment in children exposed during pregnancy to oseltamivir. We performed an uncontrolled prospective cohort study that evaluated pregnancies with exposure to the H1N1 Influenza virus and its treatment with Oseltamivir Phosphate. The sample consisted of 589 pregnant women with suspected symptoms of Influenza A who were reported in the Information System for Notifiable Diseases - Influenza (SINAN-Influenza, Rio Grande do Sul Database). Follow-up of 424 pregnancies was conducted via telephone, home visit, medical records or Live Birth Certificate, by a trained team. PCR (polymerase chain reaction) was performed in 243 individuals. There were 163 (67%) confirmed cases of H1N1 and 80 (33%) non-H1N1 Influenza virus. There were twenty-four maternal deaths, 18 of these were H1N1+ patients. Eight stillbirths were reported, five of these were for H1N1+ pregnant women. There were no differences in perinatal outcomes. Only one cleft palate was reported in a newborn whose mother did not use oseltamivir. Use of oseltamivir phosphate was identified in 221 patients. Of this, there were 86 confirmed cases of Influenza A (H1N1) and 51 non-H1N1 Influenza virus. Adverse reactions were reported in 92 (42%) pregnancies. There were a higher number of adverse effects reported in non-H1N1 patients after the use of oseltamivir. There were fewer maternal deaths (7.2%) in those who received oseltamivir compared to 34.7% of women who were not treated (OR: 0.14, CI95%: 0.04-0.42, p=0.0003). Similarly, the frequency of stillbirth was lower (2.2%) in treated as compared to 13.0% of the untreated women (OR: 0.15, CI95%: 0.03- 0.89, p=0.03). Developmental delay in two or more skills was reported in 10 (19.2%) of 52 children exposed prenatally to oseltamivir and followed for at least 36 months. This rate is above of expected for the Brazilian population (15%). In conclusion, it is expected that this work can contribute to a better understanding towards the potential teratogenic effect of Influenza A (H1N1) virus and its treatment with oseltamivir. Future studies will be decisive to the establishment of clinical practices about treatment and general management of this condition in this specific group of patients.

Dissertação (mestrado)—Universidade de Brasília, Faculdade de Medicina, Núcleo de Medicina Tropical, 2013. / Submitted by Larissa Stefane Vieira Rodrigues (larissarodrigues@bce.unb.br) on 2014-11-06T18:36:45Z No. of bitstreams: 1 2013_LíbiaRobertaDeOliveiraSouza.pdf: 1103965 bytes, checksum: 1dbd08ad6319a90b02749362cb1da2a9 (MD5) / Approved for entry into archive by Raquel Viana(raquelviana@bce.unb.br) on 2014-11-06T19:09:48Z (GMT) No. of bitstreams: 1 2013_LíbiaRobertaDeOliveiraSouza.pdf: 1103965 bytes, checksum: 1dbd08ad6319a90b02749362cb1da2a9 (MD5) / Made available in DSpace on 2014-11-06T19:09:48Z (GMT). No. of bitstreams: 1 2013_LíbiaRobertaDeOliveiraSouza.pdf: 1103965 bytes, checksum: 1dbd08ad6319a90b02749362cb1da2a9 (MD5) / Em abril de 2009 foram identificados os primeiros casos por Influenza A(H1N1)pdm09 e, em junho desse mesmo ano, foi declarada fase pandêmica por este novo subtipo. As gestantes foram identificadas precocemente como grupo de risco para complicações e óbito. O Brasil apresentou decréscimo de 51% na morte materna no período de 1990 e 2010, porém houve um incremento nesse indicador no ano de 2009, o qual pode ter sido influenciado por tal pandemia. O objetivo desse estudo foi descrever a mortalidade por Influenza A(H1N1)pdm09 entre gestantes no Brasil, nos anos de 2009 e 2010. Foi realizado um estudo descritivo sobre a mortalidade em gestantes empregando-se o método de relacionamento probabilístico entre os óbitos por todas as causas no Sistema de Informação sobre Mortalidade e as notificações no Sistema de Informação de Agravos de Notificação em mulheres de 10 a 59 anos. Foram realizados 7 passos de blocagem e as variáveis nome e data de nascimento foram utilizadas para o pareamento. Foram identificados óbitos subnotificados e subinformados no SIM e calculada a proporção de mortes maternas por Influenza A(H1N1)pdm09. Foram notificadas 6.695 gestantes no período estudado e confirmados 3.301 (49,3%) casos por Influenza A(H1N1)pdm09. Em 2009, 52,% (3.164/6.000) dos casos foram confirmados, o equivalente a 95,8% no período de estudo. Dentre o total de gestantes notificadas, 371 (5,5%) evoluíram para óbito, sendo 61,2% (227) dos óbitos confirmados para Influenza A(H1N1)pdm09. Enquanto menos de 3% da população de mulheres em idade fértil era gestante, mais de 30% das mulheres em idade fértil, que evoluíram para óbito por Influenza A(H1N1)pdm09 eram gestantes. Quarenta e dois por cento dos óbitos tinham registro de pelo menos uma comorbidade e mais de 50% estavam no terceiro trimestre da gestação. O pico das mortes maternas pelo grupo das doenças do aparelho respiratório complicando a gravidez, parto e puerpério coincidiu com o pico dos óbitos em gestantes por influenza A(H1N1)pdm09. Após identificação dos óbitos subnotificados e subinformados em gestantes por Influenza A(H1N1)pdm09, foi evidenciado que, entre 9,9% e 9,0% das mortes maternas, em 2009, e entre 1,8% e 1,5%, em 2010, foram por Influenza A(H1N1)pdm09. Excluindo as mortes maternas por Influenza A(H1N1)pdm09, a Razão de Morte Materna não sofreria incremento em 2009. A pandemia pelo vírus Influenza A(H1N1)pdm09 foi responsável pelo aumento de morte materna em 2009, no Brasil, evidenciado por exagerada representação de gestantes entre os óbitos por Influenza A(H1N1)pdm09 nas mulheres em idade fértil e por elevação do número de mortes maternas. Recomenda-se que sejam realizados estudos sobre o uso de oseltamivir e da vacina contra influenza na mitigação dos óbitos por influenza em gestante. _______________________________________________________________________________________ ABSTRACT / In April 2009, the first cases of Influenza A(H1N1)pdm09 were identified and in June of that year was declared pandemic phase for this new subtype. Pregnant women were at increased risk of complications and death according to early reports on the pandemics epidemiology. Brazil had a 51% decrease in maternal deaths between 1990 and 2010, but there was an increase in this indicator in 2009, which may have been influenced by such a pandemic. The aim of this study was to describe mortality from Influenza A(H1N1)pdm09 among pregnant women in Brazil during the years 2009 and 2010. We conducted a descriptive study on mortality among pregnant women using the method of probabilistic record linkage between deaths from all causes in the Mortality information System (SIM) and reports of cases in women with 10-59 years-old. Seven blocking steps were executed and variables name and date of birth used for pairing. We identified underreported and under informed deaths in the SIM and calculated the proportion of maternal deaths due to Influenza A(H1N1)pdm09. During the study period, 6,695 pregnant women were reported and 3,301 (49.3%) were confirmed for influenza A(H1N1)pdm09. In 2009, 52% (3.164/6.000) of the cases were confirmed, equivalent to 95.8% in 2009-2010. Among the total number of pregnant women reported, 371 (5.5%) died, among then 61.2% (227) was confirmed as influenza A(H1N1)pdm09 deaths. While less than 3% of the population of women of childbearing age was pregnant, over 30% of them who have died due to Influenza A(H1N1)pdm09 were pregnant. Forty- two percent of recorded deaths had at least one known comorbidity, and over 50 % were in the third trimester of pregnancy. The peak of maternal deaths ordered by group of respiratory diseases complicating pregnancy, childbirth and puerperium coincided with the peak of deaths in pregnant women with influenza A(H1N1)pdm09. After identification of underreported and under informed deaths among pregnant women by Influenza A(H1N1)pdm09, it was shown that between 9.9% and 9.0% of maternal deaths in 2009 and between 1.8 % and 1.5 % in 2010 were due to Influenza A(H1N1)pdm09. Excluding maternal deaths due to Influenza A(H1N1)pdm09, the Maternal Mortality Ratio in Brazil did not suffer an increase in 2009. The pandemic of Influenza A(H1N1)pdm09 was responsible for the increase of maternal death in Brazil in 2009, evidenced by overrepresentation of deaths among pregnant women with Influenza A(H1N1)pdm09 in women of childbearing age and increased number of maternal deaths. It is recommended that studies on the use of oseltamivir or the influenza vaccine in mitigating influenza deaths in pregnant women.