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Despite decades of research about educational interventions to correct parents' childhood fever management their knowledge remains poor and practices continue to be based on beliefs about harmful outcomes. The purpose of this thesis was to 1) identify Australian parents' fever management knowledge, attitudes, practices and methods of learning to manage fever and 2) undertake a theoretical exploration of the determinants of parents' intentions to reduce fever using the Theory of Planned Behavior (TPB). Two studies were undertaken: a qualitative study with 15 parents; and survey of 401 Queensland parents with a child aged between 6 months and 5 years. Parents determine childhood fever through behavioural changes they have learnt to associate with fever. Few were aware of the immunological beneficial effects associated with fever and most believed fever harmful causing febrile convulsions and brain damage. To prevent harm they monitored temperatures, used antipyretics, dressed children in light clothing and sponged them with tepid, cool or cold water. Despite believing antipyretics harmful most parents reduced temperatures of 38.3 degrees Celsius ± 0.6 degrees Celsius with antipyretics, alternating two antipyretics when fever was not reduced or returned. In addition to temperature reduction antipyretics were used to reduce distress or general unwellness and pain or discomfort. Multiple factors were used to determine antipyretic dosage including temperature, irritability and illness severity. Over one-third of parents had overdosed their child with too frequent antipyretic administration; more frequently with ibuprofen than paracetamol, 12:1. Fever management information was learnt from numerous sources. Doctors were the most frequently reported followed by personal experience. With the variety of information sources nearly half received conflicting information about how to manage fever increasing concerns and creating uncertainty about how to best care for their child. Despite this many believed they knew how to manage fever. Some parents' practices changed over time as a result of either positive or negative experiences with fever indicating more positive or negative attitudes toward fever. Positive experiences reduced antipyretic and medical service use; negative ones had the adverse effect with increase in antipyretic use including alternating antipyretics and double dosing with one antipyretic. Child medication behaviours also influenced attitudes and practice intentions. Parents of children who readily took antipyretics had more negative attitudes and intended to reduce their child's next fever with antipyretics. Negative attitudes were a significant determinant of fever management intentions. Parents' practices were strongly influenced by their perception that doctors and partners expected them to reduce fever. This expectation from partners is understandable; from doctors it is not and indicates doctors' propensity to recommend reducing fever. There is an urgent need to identify doctors' fever management beliefs and rationales for practice recommendations. Parents also learn to manage fever from nurses and pharmacists; their beliefs and management rationales must also be determined and addressed. There is an urgent need to educate parents about evidence-based fever management and reduce their unnecessary antipyretic use. They must be encouraged to delay antipyretic administration using them to reduce pain rather than fever. Findings from this thesis have identified the determinants of parents' intentions to reduce fever; negative attitudes and normative influences and positive child medication behaviours. Future studies should examine the efficiency and cost effectiveness of fever management educational programs for parents using different presentation methods in multiple settings.

Submitted by Luiz Cesar Maranho (lc-maranho@bol.com.br) on 2018-10-11T20:16:50Z No. of bitstreams: 1 Dissertação Final.pdf: 1883342 bytes, checksum: 85a25606a3113b39d6d4354dcaa161d8 (MD5) / Approved for entry into archive by Elza Mitiko Sato null (elzasato@ibilce.unesp.br) on 2018-10-15T12:39:20Z (GMT) No. of bitstreams: 1 maranho_lc_me_sjrp.pdf: 5069791 bytes, checksum: 2501e6acc67bdd7103eb807f326a4c0b (MD5) / Made available in DSpace on 2018-10-15T12:39:20Z (GMT). No. of bitstreams: 1 maranho_lc_me_sjrp.pdf: 5069791 bytes, checksum: 2501e6acc67bdd7103eb807f326a4c0b (MD5) Previous issue date: 2018-08-20 / O mosquito Aedes aegypti é o principal vetor responsável por diversas arboviroses como a dengue, a febre amarela, o vírus zika e a febre chikungunya. Devido a sua resistência, adaptabilidade e proximidade ao homem, o Aedes aegypti é atualmente um dos maiores problemas de saúde pública no Brasil e nas Américas. Mesmo com os avanços e investimentos em pesquisas com vacinas, monitoramento, campanhas educativas e diversos tipos de controle deste vetor, ainda não existe um método eficaz para controlar e erradicar o mosquito. Portanto, esse trabalho destina-se ao auxílio na criação de estratégias para controlar esse agente transmissor, mediante a análise do espaço de estados e a estabilidade assintótica de uma dinâmica não linear para controle populacional do Aedes aegypti via a técnica de linearização de Lyapunov, além de apresentação de formas de prevenção e combate aos criadouros do mosquito. A dinâmica não linear proposta é uma dinâmica simplificada obtida de um modelo não linear existente na literatura, proposto por Esteva e Yang em 2005 e se baseia no ciclo de vida do mosquito, que é dividido em duas fases: fase imatura ou aquática (ovos, larvas e pupas) e fase alada (mosquitos adultos). Na fase adulta, os mosquitos são divididos em machos, fêmeas imaturas e fêmeas fertilizadas, sendo que a dinâmica proposta nesta dissertação de mestrado é baseada nos estudos efetuados por Reis desde 2016, obtendo um modelo simplificado no qual a soma das densidades das populações de fêmeas imaturas e fêmeas fertilizadas serão consideradas como fêmeas adultas. / The mosquito Aedes aegypti is the main vector responsible for several arboviruses such as dengue fever, yellow fever, zika virus and chikungunya fever. Due to its resistance, adaptability and proximity to humans, Aedes aegypti is currently one of the major public health problems in Brazil and the Americas. Even with the advances and investments in research with vaccines, monitoring, educational campaigns and various types of control of this vector, there is still no effective method to control and eradicate the mosquito. Therefore, this work is intended to aid in the creation of strategies to control this transmitting agent by analyzing the state space and the asymptotic stability of a nonlinear dynamics for population control of Aedes aegypti via the Lyapunov linearization technique to present ways of preventing and combating mosquito breeding sites. The proposed nonlinear dynamics is a simplified dynamics obtained from a nonlinear model existing in the literature, proposed by Esteva and Yang in 2005 and based on the life cycle of the mosquito, which is divided into two phases: immature or aquatic phase (eggs, larvae and pupae) and winged phase (adult mosquitoes). In the adult phase, mosquitoes are divided into males, immature females and fertilized females, and the dynamics proposed in this dissertation is based on studies carried out by Reis since 2016, obtaining a simplified model in which the sum of the densities of the populations of females immature and fertilized females will be considered as adult females.

Hantaviruses are members of the virus family Bunyaviridaethat cause two potentially life-threatening diseases in humans: hemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (BPS). HFRS is caused by Old World hantaviruses that are endemic in many Asian and European countries. Infections with Old World hantaviruses can range in severity from asymptomatic to moderate or severe, depending primarily on the infecting serotype of virus. HPS is caused by New World hantaviruses in North and South America. New World hantaviruses are rarely asymptomatic and are severe in the majority of cases. These syndromes are distinct from one another in the primary target organ of virus infection (kidney vs. lung), but have important clinical features in common, including fever, thrombocytopenia, and a capillary leak syndrome. These common clinical manifestations suggest that the underlying mechanisms of disease may be similar in the two syndromes. The precise mechanisms of pathogenesis of HFRS and HPS are poorly characterized, but may be mediated in part by immunopathology. Hantaviruses are able to establish infections in many human cell types, including primary human endothelial cells, without having any cytopathic effect on these cells. Human infections with hantavirus result in a robust activation of the humoral and cellular immune response, and we hypothesize that these immune responses contribute to the pathology of disease. Evidence for the activation of T lymphocytes, and their potential involvement in immunopathology, includes increases in the number of circulating, activated CD8+ T cells during HFRS, the presence of lymphocytic infiltrates (predominantly CD8+T cells) in kidney biopsies from patients with acute HFRS, and associations between certain HLA haplotype and disease severity following hantavirus infection. This thesis is the first examination of human T lymphocyte responses that are generated during HFRS. Initially, we studied memory T cell responses in scientists who were sub-clinically infected with Hantaan virus (HTNV), the prototype hantavirus. We later investigated memory T cell responses in healthy Finnish adults who had HFRS caused by Puumala virus (PUUV), a hantavirus endemic primarily in Scandinavia. At the onset of these studies, there was no available information on human T lymphocyte responses to Old World hantaviruses. Virus-specific CD8+ and CD4+human T cell lines had been isolated from patients with acute HPS caused by Sin Nombre virus (SNV) infection. In that study, conducted in our laboratory, several human T cell epitopes on the nucleocapsid (N) protein and G2 envelope glycoprotein of SNV were identified and characterized. We decided to perform similar analyses on PBMC from donors who had been infected with HTNV and PUUV, in order to determine the specificity and diversity of the T cell response to Old World hantaviruses. The initial study of three donors who had sub-clinical infections with HTNV demonstrated that virus-specific T cell responses could be detected in all the donors following in vitro stimulation of PBMC with inactivated virus. In two of the donors, the virus-specific cytolytic T cells (CTL) recognized the HTNV N protein, and in the third donor the virus-specific CTLs recognized the HTNV G1 glycoprotein. Isolation and characterization of virus-specific T cells from two donors resulted in the identification of two CD8+ T cell epitopes on the HTNV N protein, which were restricted by either HLA A1 or B51. These CTL lines included both HTNV-specific (HLA B51-restricted) and serotype-cross reactive (HLA A1 restricted) lines. In one subject, these virus-specific T cell responses were detectable in IFN-γ ELISPOT assays following peptide stimulation, and in bulk cultures after short-term stimulation with inactivated HTNV. These results indicated that the CD8+CTL responses of humans after sub-clinical infection with HTNV were readily detectable and were directed against a limited number of viral proteins and epitopes. In addition, sub-clinical infection resulted in the generation of both virus-specific and cross-reactive CTL responses. We reasoned that hantavirus infections that lead to clinical illness may result in the generation of more robust and/or diverse virus-specific T cell responses than in sub-clinical infections. To address this question, we studied the memory CD8+ T cell responses in a group of healthy adults from Finland who had HFRS caused by PUUV infection between the years 1984 and 1995. We detected virus-specific CTL in the bulk cultures of seven of eleven immune individuals tested following stimulation with infectious virus. The PUUV proteins N, G1 and G2 were recognized by CTLs in six, five, and two donors respectively. Extensive cloning of T cells from two donors resulted in the isolation of sixty-three virus-specific CTL lines, the majority of which (61/63) were specific for the PUUV N protein. Six novel CD8+ CTL epitopes and one CD4+ CTL epitope were identified on the N protein, all of which clustered in the center of the protein between amino acids 173 and 251. The CTL lines specific for these epitopes were restricted by a variety of HLA alleles including A2, A28, B7 and B8, and were primarily serotype specific when tested against target cells expressing HTNV or SNV N protein. IFN-γ ELISPOT analysis using the defined epitopes to stimulated PBMC, revealed high frequencies of circulating N-specific CD8+ T cells in eight of thirteen individuals tested. Finally, T cell receptor (TCR) Vβ analysis of CTL clones specific for one epitope (N204-12) demonstrated that cells in this population expressed up to five different Vβ chains. These results demonstrated that the PUUV N protein may be the dominant target of the CTL response, that the N-specific CD8+ CTL responses are diverse, heterogeneous, and primarily serotype specific, and that virus-specific memory CD8+T cells can persist at high levels for up to 15 years after the primary infection. In order to understand the pathology of HFRS and HPS, we must be able to assess the contribution of various factors that could potentially contribute to disease. The virus burden in the infected individual is likely to be an important factor in the severity of the resulting disease. Quantitative RT-PCR analysis of plasma samples from acute HPS patients demonstrated that a higher virus burden (as reflected by viral RNA copy number) is associated with more severe HPS. In order to perform similar analyses in patients with HFRS caused by PUUV, we established a quantitative RT-PCR assay for the detection of PUUV S segment RNA in patient plasma. The design and optimization of the PUUV-specific RT-PCR is described in this report. This assay will allow us to measure the virus burden in patients and compare these data with levels of T cell activation and with parameters of disease severity. In this way, we hope to gain an understanding of the kinetics and magnitude of both the virus burden and virus-specific T cell response during the acute illness. This thesis provides the first description of human virus-specific T cell responses to HTNV and PUUV. These data shed light on the nature of the CD8+ T cell responses that are generated following natural infections with PUUV and sub-clinical infections with HTNV. The studies of memory CD8+ T cell responses to PUUV, and the development of a PUUV-specific quantitative RT-PCR assay, establish the framework for future studies of the immunopathology of acute HFRS. Quantitative analysis of both virus burden and T cell responses during acute illness will provide insight into their relative contributions to the pathology of disease.

The main goal of thesis is to explore the changing global epidemiological situation depending on climate changes and to map trends of changes in epidemiological situation. A sub-objective is to find out if there are differences in the geographical distribution of the incidence of West Nile Virus (WNV) in the period 2004-2014 next to map surveillance, to propose options for improving prevention in a changing geographical distribution, to describe cases of the diseases reported in the Czech Republic, to determine whether in the Pilsen and Budweis medical facilities are routinely investigating also on detection of WNV infection and finally to chart the WNV infection rate and its causes in horse breeding. Influence of climate change on distribution of WNV can be very well observed. This change is mainly due to settlement of new countries by the originator and also reservoirs which are moving further to the north or south becouse of climate. Distribution of WNV to the new countries is also supported by the changing of landscape, mainly due to the changing climate. The changes of migration routes of migratory birds which are also affected by climate changes have also the effect on changing geographic distribution. The globalization of the world is also mostly responsible for change of distribution. The change of epidemiological situation is given by that the virus attacks organisms that did not meet with virus until now. Therefore, there is a change in the clinical picture and more serious forms of the disease appear in the greater degree than before. Official statistics numbers of diseases in the EU in 2008-2012 published by the ECDC revealed that number of illnesses depends on the global climate changes and also on the climate in a current year. Data for year 2010 clearly reveal how big influence extreme temperatures have on the number of reported illnesses. With regard to prevention in the Czech Republic, as a main step was to designed the informing of tourists. The substantial part of the repressive measures against WNV disease is to stop transmission of WNV blood transfusions. This deals with in annex 3of Decree no. 143/2008 Coll.. Currently the draft of methodological instruction which sets out the procedure for assessing the risk of WNV infection and the procedure to reduce the risk of transmission of the virus through blood transfusions is in the comment procedure. By the survey of health facilities was found out that it is according the legislation, specifically Decree 233/2011 Coll.. Private microbiology and serology laboratories in Pilsen and Budweis do not test on positivity of WNV. Targeted surveillance of horse WNV in Czech Republic is doing from 2011. In 2011 2013 it was 4 5 WNV positive horses, in 2014 it was already 13 horses from a total of 783 horses tested positive WNV. Various geographic distribution of horses and high specific antibody titers according to veterinarians indicate increasing activity of the virus in the Czech Republic and the possible expansion of WNV into new areas. The thesis also analyzes three cases of WNV disease that have been imported to the Czech Republic from the USA, Tanzania and from Cyprus. In thesis is also described the first autochthonous case of WNV infection in the Czech Republic in 2013 which demonstrates the changing epidemiological situation in the country. Based on the obtained data is appreciable that the virus will be with regard to a changing climate and increased globalization spread to new areas in which will be probably cause diseases with serious clinical course. Whereas the virus has a tends to mutation it is expected also a gradual change of the clinical picture. To stop the spread of the virus should be carried out surveillance at all levels. Insomuch as is not in current time human vaccine available to curb the spread of the The essential elements od prevention includes informing tourists traveling to high-risk countries and countries potentially risky.

Submitted by Tereza Risi (tereza.risi@icict.fiocruz.br) on 2013-10-11T19:29:13Z No. of bitstreams: 1 Isabel Levy.pdf: 2081227 bytes, checksum: d87790882fbeedaf43096403d4e742be (MD5) / Made available in DSpace on 2013-10-11T19:29:13Z (GMT). No. of bitstreams: 1 Isabel Levy.pdf: 2081227 bytes, checksum: d87790882fbeedaf43096403d4e742be (MD5) Previous issue date: 2013 / Fundação Oswaldo Cruz. Instituto de Comunicação e Informação Científica e Tecnológica em Saúde. Rio de Janeiro, RJ, Brasil / O advento e a popularização das tecnologias da informação e da comunicação ± dentre elas, a internet e as redes sociais on - lin e ± têm modificado profundamente as formas como pessoas e instituições se relacionam. No campo da Comunicação e Saúde, diversas experiências contemporâneas apontam a concorrência entre forças centrípetas e centrífugas no ciberespaço e levantam a questão: o investimento das instituições públicas de saúde em atividades desenvolvidas na internet, por meio das redes sociais on - line, é de fato uma inovação em suas práticas discursivas ou corresponde a uma inovação relativa ao suporte, mas não ao modo de relação com os seus interlocutores? Por meio da abordagem da netnografia, a pesquisa buscou mapear, descrever e analisar práticas de comunicação do Ministério da Saúde no ciberespaço, tendo como objeto específico a Campanha Nacional de Combate à Dengue 2011/2012. O estudo aponta que, a o assumir algumas características que tradicionalmente são observadas na forma como a mídia hegemônica aborda os temas de saúde, os espaços virtuais do Ministério da Saúde constituem - se mais como mídias digitais e menos como redes soc iais on - line. Os resultados nos levam a concluir que, apesar da inovação no uso de suportes digitais e no investimento em recursos criativos como as narrativas transmídia, as práticas de Comunicação e Saúde desenvolvidas pelo Ministério da Saúde no ciberes paço durante a Campanha estudada repetem as dinâmicas que caracterizam o modelo hegemônico de comunicação de natureza transferencista, bipolar, linear unidirecional e centralizado no emissor, e consolidam o discurso campanhista de saúde / The information and communication technologies ± especially the internet and online social networks - have profoundly changed the relationshi p between people and institutions. In the field of Communication and Health, various contemporary experiences indicates the competition between centripetal and centrifugal forces in cyberspace and raise the question about the public institutions investmen t in online activities. Is it is indeed an innovation in their discursive practices or represents an innovation on the devices , but not on the mode of relationship with their interlocutors? Through netnography approach, the research sought to identify, des cribe and analyze communication practices of the Ministry of Health in cyberspace, with the specific object of the National Campaign to Combat Dengue 2011/2012. The study shows that by taking a few features that are traditionally found in how the hegemonic media covers WKH WRSLFV RI KHDOWK 0LQLVWU\ RI +HDOWK¶V YLUWXDO VSDFHV DUH PRUH OLNH GLJLWDO PHGLD DQG less like online social networking. The results lead us to conclude that, despite the innovative use of digital media and creative resources as transmed ia narratives, Communication and Health practices developed by the Ministry of Health in cyberspace during the Campaign repeat the dynamics that characterize the hegemonic model of communication - bipolar, linear and unidirectional, centralized in an emit ter polo - and consolidate the health's prescriptive speech

Made available in DSpace on 2011-11-09T14:45:44Z (GMT). No. of bitstreams: 2 license.txt: 1648 bytes, checksum: e095249ac7cacefbfe39684dfe45e706 (MD5) 000239.pdf: 1746615 bytes, checksum: b1538d716409c5efb4a08da0bc08f73d (MD5) Previous issue date: 2008 / Fundação Oswaldo Cruz. Instituto Fernandes Figueira. Departamento de Ensino. Programa de Pós-Graduação em Saúde da Criança e da Mulher. Rio de Janeiro, RJ, Brasil / Objetivo: avaliar a evolução clínica, morbidade e mortalidade de crianças e adolescentes portadores de cardiopatia reumática em acompanhamento ambulatorial num centro terciário. Material e Métodos: estudo descritivo observacional longitudinal de base hospitalar. Foi realizada análise de prontuários de pacientes com 3-18 anos, acompanhados por no mínimo 2 anos no ambulatório do Instituto Nacional de Cardiologia no Rio de Janeiro. O diagnóstico foi confirmado pelos critérios de Jones e/ou exame ecocardiográfico com lesão reumática típica mitral e/ou aórtica. Banco de dados foi elaborado com o programa ACCESS 2000, e a análise estatística realizada com o programa EPI-INFO 2000. Foi considerado significativo o valor de α- 0,05. Resultados:139 prontuários foram incluídos no estudo. A mediana da idade no início do seguimento foi de 11 anos, 52,6% eram do sexo feminino. Quanto à forma de apresentação clínica 45,3% estavam no primeiro surto, 14,4% em recidiva e 40,3% na fase crônica. A mediana de idade dos pacientes crônicos e em recidiva era superior aos pacientes do primeiro surto de febre reumática (p-0,0001). O tempo médio de seguimento foi de 9,9 anos (2-21 anos). A lesão valvar predominante foi a insuficiência mitral (82,7%), seguida da insuficiência aórtica (55,9%) e da insuficiência mitro-aórtica em 45,3% dos casos. Houve redução importante dos percentuais de lesões valvares graves - tanto mitrais como aórticas - ao final do seguimento.Recidivas foram evidenciadas em 32,3%. Pacientes com profilaxia irregular ou sem profilaxia apresentaram a média do número total de surtos (2,4 surtos por paciente) superior a do grupo em profilaxia regular (1,4 surtos por paciente), com diferença entre os grupos estatisticamente significante (p-0,0009).A mortalidade foi de 4,3% (n=6) Todos os pacientes que evoluíram para óbito eram portadores de próteses valvares. O abandono de tratamento foi de 10,8%, sendo que 1,4% desses pacienteseram portadores de próteses mecânicas (n-=2). Procedimentos intervencionistas foram realizados em 45,3% dos pacientes - valvuloplastia por cateter balão em 2,9% e cirurgia cardíaca valvar em 42,4%. Reoperação foi necessária em 8,6% (2ª cirurgia) e 2,8% (3ª cirurgia). O procedimento mais realizado foi o implante de prótese mecânica mitral (31,3%), seguido por prótese mecânica aórtica (20,9%) e plastia mitral (18,6%). A endocardite infecciosa foi evidenciada em 8,6%, sendo a endocardite de prótese em 3,6%, responsável por 50% da mortalidade desta amostra, com letalidade de 25%. Outrascausas de mortalidade incluíram estenose grave de prótese biológica (n=1), estenose grave de prótese mecânica (n=1) e trombose de prótese biológica (n=1). / Objective: to investigate the outcome, mortality and morbidity of children and adolescents with rheumatic heart disease followed up in an outpatient care unit of a terciarie center. Methods– descriptive longitudinal observational study of an hospitalar based population. Medical file of patients – 3 to18 years old - with rheumatic heart disease, followed-up for at least 2 years from in the outpatient care unit of the National Cardiology Institute (InstitutoNacional de Cardiologia) in Rio de Janeiro were reviewed. Diagnosis were confirmed through medical file register of the revised Jones criteria for rheumatic fever and/or Doppler echocardiographic report of typical chronic mitral or aortic lesions. Database program ACCESS 2000, statistical analysis was performed using EPI-INFO 2000 software, with significant αvalue 0,05. Results–139 medical files were reviewed. Median age at the first visit to the service was 11 years, 52,5% were female. At the first exam, 45,3% presented with acute rheumatic fever- first attack, 14,4% recurrence; while 40,3% had chronic valvular lesions. Median age of the chronic and recurrent group was greater than median age of the first attack group (p-0,0001). Mean follow-up time was 9,9 years (2 to 21 years). Mitral regurgitation was the most common valvular lesion (82,7%), followed by aortic regurgitation (55,9%) and combined mitral and aortic regurgitation (45,3%). There was a significant percent decrease in severe valvular lesions - both mitral and aortic – at the end of follow-up period. Recurrences were present in 32,3% of cases. There was a significant difference (p-0,0009) between the mean rate of the total number of attacks of patients under irregular or no prophylaxis (2,4 attacks / patient) compared with patients under regular prophylaxis (1,4 attacks / patient). Mortality rate were 4,3% (n=6). All these patients that died had prosthesis. 10,8% were lost of follow-up - 1,4% of these patients had mechanical prostheses (n=2). 45,4% underwent valve procedures: 2,9% balloon dilatation and 42,4% valve surgery. Reoperation wererequired by 8,6% (2 nd surgery) and rereoperation by 2,8% (3 rd surgery). The most common surgical procedure was mitral valve replacement with mechanical prosthesesimplantation (31,3%), followed by aortic valve replacement with mechanical prostheses implantation (20,9%) and mitral valve repair (18,6%). A total of 8,6% presented with endocarditis - 3,6% had prosthetic valve endocarditis, that accounted for 50% mortality of this group, and for a letality rate of 25%.Another causes of death included severe bioprosthesis stenosis (n=1), severe mechanical prosthesis stenosis (n=1) and bioprosthesis valve trombosis (n=1).

Submitted by Priscila Nascimento (pnascimento@icict.fiocruz.br) on 2012-11-16T12:24:56Z No. of bitstreams: 1 rafael-lawson-ferreira.pdf: 3259100 bytes, checksum: 120506960b3b1958e6509f56db6fef38 (MD5) / Made available in DSpace on 2012-11-16T12:24:56Z (GMT). No. of bitstreams: 1 rafael-lawson-ferreira.pdf: 3259100 bytes, checksum: 120506960b3b1958e6509f56db6fef38 (MD5) Previous issue date: 2007 / Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos. Rio de Janeiro, RJ, Brasil / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil. / O Instituto de Tecnologia em Imunobiológicos - Bio-Manguinhos/ FIOCRUZ (BM) é um importante centro de produção de imunobiológicosna América Latina. Dentre as vacinas produzidas em Bio-Manguinhos, sob condiçõesde boas práticas de fabricação, a vacina contra Febre Amarela (VcFA), produzida a partir de ovos embrionados SPF, é certificada pela Agência Nacional de Vigilância Sanitária e Organização Mundial de Saúde para suprir a demanda do Ministério da Saúde e das Agências das Nações Unidas. Para garantir a segurança da vacina assim como a ausência de agentes adventícios, testes de Controlede Qualidade (CQ) devem ser realizados na matéria-prima e nas diferentes etapasda produção. Por recomendação da Organização Mundial de Saúde, em vacinas produzidas para o uso humano deve ser demonstrada a ausência de M. oralee M. pneumoniaee quando material de origem aviária é utilizado durante a produção, ausência de M. gallisepticume M. synoviae. Da mesma forma como o desenvolvimento de novos produtos tem evoluído, o CQ deve seguir estas novasabordagens cujos benefícios imediatos seriam a rapidez na liberação de resultados, maior sensibilidade e especificidade. O micoplasma é um importante agenteadventício amplamente encontrado em diferentes tipos de substratos biológicos, porém é um microrganismo fastidioso e de difícil detecção pelo exame direto (cultura do microrganismo) o qual requer um longo prazo para obtenção de resultados, cerca de 35 dias. Em nosso trabalho selecionamos o par de oligonucleotídeos iniciadores GPO-1 e MGSO com o objetivo de estabelecer um teste para detecção de micoplasmas por PCR no CQ da VcFA. Os oligonucleotídeos selecionados amplificam fragmentos de aproximadamente 700 pb na região do gene do rDNA 16S. Foram testadas diferentes metodologias de extração de DNA, como fervura, fenol/clorofórmio e kits comerciais avaliando a adaptabilidade da técnica para os diferentes produtos intermediários. A metodologia proposta, neste trabalho, foi capaz de detectar M. pneumoniaeem concentrações entre 6,25 e 3,125 UFC/mL e M. synoviaeentre 12,5 e 6,25 UFC/mL, em produtos intermediários intencionalmente contaminados, em concentrações inferiores do que as preconizadas pela OMS. Além disso, os oligonucleotídeos selecionados demonstraram especificidade pelas espécies de micoplasmas utilizadas, não apresentando amplificação quando o DNA de outras espécies bacterianas foi utilizado. Paralelamente, foi realizado um ensaio de polimorfismo do comprimento dos fragmentos da restrição (RFLP) para identificação da espécie de micoplasma, onde selecionamos as enzimas de restrição MboI e HinfI, capazes de diferenciar as três espécies de micoplasmas de referência utilizadas (M. gallisepticum, M. pneumoniaee M. synoviae). / The Institute of Technology in Immunobiologicals - Bio-Manguinhos/ FIOCRUZ is an important immunobiological production center in Latin America. Among the vaccines produced in Bio-Manguinhos under good manufacturingpractice, the vaccine against Yellow Fever (YF-Vaccine), produced in SPF embrionated eggs, is certified by the Brazilian National Surveillance Agency and World Health Organization to achieve the demand of the Brazilian Ministry of Health and the United Nations Agencies. In order to guarantee the safety of the vaccine as well as the absence of contaminant agents, tests of Quality Control (QC) must be done along its production. By advice of WHO, on vaccines produced for human use, there must be demonstrated the absence of M. oraleand M. pneumoniae. When poultry material is used during the production, absence of M. gallisepticumand M. synoviae is also required. Mycoplasma is a fastidious microorganism of difficult detection through culturing, which requires a long term for conclusive results, about 35 days. We selected the primer pair GPO-1 and MGSO to establish a mycoplasma detection through PCR in YF-Vaccine. The selected oligonucleotides amplify fragments of roughly 700 bp in the 16S rRNA gene region. Different methodologies of DNA extraction were performed, such as boiling, phenol/chloroform and commercial kits, thus, evaluating the adaptability of each method for the intermediary products. The proposed methodology in this study was capable to detect M. pneumoniaein concentration between 6,25 and 3,125 CFU/mL and M. synoviae,between 12,5 and 6,25 CFU/mL, in spiked intermediate products which constitute lower detection limits throw thosestipulated by WHO. In addition, selected oligonucleotides demonstrated specificity to the Mycoplasma species used in this work, not presenting amplification productswhen other bacteria species was used. Three reference mycoplasmas species (M. gallisepticum, M. pneumoniaeand M. synoviae) could be differentiated based on restriction fragments length polymorphism (RFLP) after digestion of the amplification products by HinfI and MboI.

Submitted by Priscila Nascimento (pnascimento@icict.fiocruz.br) on 2013-03-19T16:07:54Z No. of bitstreams: 1 Ana_Carolina.pdf: 2560132 bytes, checksum: 663a398860ff953a4b1de21a209ee151 (MD5) / Made available in DSpace on 2013-03-19T16:07:54Z (GMT). No. of bitstreams: 1 Ana_Carolina.pdf: 2560132 bytes, checksum: 663a398860ff953a4b1de21a209ee151 (MD5) Previous issue date: 2012 / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil. / O desenvolvimento de uma vacina para malária é considerado atualmente uma prioridade em saúde pública pelo impacto socioeconômico e morbidade da doença com 250 milhões de novos casos por ano. A vacina de febre amarela é considerada uma das vacinas mais bem sucedidas por sua imunogenicidade duradora obtida após uma única dose. Frente a elucidação das respostas polivalentes dirigidas ao vírus vacinal 17D, a utilização do mesmo como vetor de expressão para antígenos heterólogos têm sido encorajada. Tendo em vista que febre amarela e malária compartilham grandes zonas endêmicas nos continentes americano e africano, a construção de uma vacina para malária baseada no vetor de febre amarela 17D se tornou uma abordagem interessante. Foram construídos dois vírus recombinantes contendo a proteína heteróloga MSP-119de P. falciparum (FA17D/MSP-119fal) e P. vivax (FA17D/MSP-119vivax) entre os genes E/NS1 de FA. Esta proteína é constituída de um fragmento de 19 kDa obtido após o processamento proteolítico da proteína de superfície do merozoíta 1 (MSP-1) durante a invasão do eritrócito e é descrita como alvo de anticorpos protetores em animais e pessoas imunes. Os vírus recombinantes foram caracterizados in vitro quanto à capacidade proliferativa em células Vero, estabilidade genética e expressão da proteína heteróloga por microscopia confocal de imunofluorescência e western blotting. A imunização de camundongos BALB/c e primatas não-humanos da espécie Saimiri sciureus foi usada para avaliar as construções quanto à imunogenicidade. Ambos os vírus foram capazes de induzir a produção de anticorpos neutralizantes contra a FA, porém em menores títulos do que os induzidos pelo vírus vacinal 17DD. A indução de anticorpos específicos para a proteína heteróloga após a imunização com os diferentes vírus recombinantes, também foi demonstrada e resultou em baixos títulos de IgG.em ambos os modelos. Os anticorpos induzidos no modelo com macacos Saimiri reconheceram a proteína nativa do parasita em hemácias infectadas por P. falciparum. No entanto, o desafio realizado neste modelo de primata não-esplenectomizado após a imunização com FA17D/MSP-119fal não gerou resultados conclusivos. Estes resultados sugerem a necessidade de aprimoramento da plataforma de expressão em busca de maior imunogenicidade. / The development of a vaccine for malaria is currently considered a priority in public health due to the socioeconomic impact and morbidity of the disease with 250 million new cases registered every year. The yellow fever vaccine is considered one of the most successful vaccines for its longlasting immunogenicity obtained after a single dose. As a result of the elucidation of the polyvalent responses directed to YF17D vaccine virus, the use of this virus as an expression vector of heterologous antigens has been encouraged. Considering that yellow fever and malaria share the major endemic areas in American and African continents, the construction of a vaccine for malaria based on the yellow fever 17D vector became an interesting approach. Two recombinant viruses containing the heterologous protein MSP-119 from P. falciparum (YF17D/MSP-119fal) and P. vivax (YF17D/MSP-119vivax) inserted between the E/NS1 genes have been constructed. This protein consists of a 19 kDa fragment obtained after proteolytic processing of the merozoite surface protein 1 (MSP-1) during invasion of erythrocytes and is described as a target for protective antibodies in animals and immune people. Recombinant viruses were characterized in vitro for their proliferative capacity in Vero cells and genetic stability and expression of heterologous protein were assessed by confocal immunofluorescence and Western blotting. Immunization of BALB/c mice and non-human primate species Saimiri sciureus allowed the evaluation of the constructions in terms of immunogenicity. Both viruses were capable of inducing neutralizing antibodies to YF, but in lower titers than those induced by the vaccine virus 17DD.The induction of specific antibodies for the heterologous protein by the different recombinant viruses was also demonstrated by low levels of IgG in both models. The antibodies induced in this monkey model bound to the native protein in parasite-infected red blood cells by immunofluorescence. The challenge carried out in after immunization of Saimiri monkeys with FA17D/MSP-119fal did not generate conclusive results. These data suggest the need to improve the platform of expression towards higher viral immunogenicity.

Submitted by Gilvan Almeida (gilvan.almeida@icict.fiocruz.br) on 2016-09-26T14:06:07Z No. of bitstreams: 2 license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) 154.pdf: 3093937 bytes, checksum: 918716b589d18e1e0f0abe0dcbcc0d98 (MD5) / Approved for entry into archive by Barata Manoel (msbarata@coc.fiocruz.br) on 2016-10-14T13:05:33Z (GMT) No. of bitstreams: 2 license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) 154.pdf: 3093937 bytes, checksum: 918716b589d18e1e0f0abe0dcbcc0d98 (MD5) / Made available in DSpace on 2016-10-14T13:05:33Z (GMT). No. of bitstreams: 2 license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) 154.pdf: 3093937 bytes, checksum: 918716b589d18e1e0f0abe0dcbcc0d98 (MD5) Previous issue date: 2013 / Fundação Oswaldo Cruz. Casa de Oswaldo Cruz. Rio de Janeiro, RJ, Brasil. / Esta tese tem como objeto a Campanha Continental para a Erradicação do Aedes Aegypti, lançada pela Organização Sanitária Pan-Americana (OSP), em 1947. As suas origens, contudo, remontam à Campanha Mundial de Erradicação da Febre Amarela, idealizada em 1914 por Wycliffe Rose,o primeiro Diretor da Comissão de Saúde Internacional (CSI) da Fundação Rockefeller (FR), e iniciada oficialmente em 1918, após o término da Primeira Guerra Mundial (19414-1918). A Campanha se desenvolveu entre as décadas de 1910 e 1930, nas América e na África tendo sido marcada por uma série de inflexões até ser reformulada nos anos da Segunda Guerra Mundial (1939-1945) e relançada, em 1947, sob os auspícios da OSP, na época dirigida por Fred Soper, um ex-funcionário da Fundação Rockefeller com uma longa trajetória de atuação na América do Sul, no combate à doenças como ancilostomíase, malária e febre amarela. Desta data até o final dos anos 1960, a meta de erradicar o vetor da febre amarela das Américas foi perseguida, com maior ou menor intensidade, por praticamente todas as Repúblicas americanas. O objetivo é analisar as origens, desenvolvimento histórico,impactos e controvérsias suscitadas por este que foi o primeiro e mais duradouro programa internacional de erradicação de uma doença já implantado. A hipótese é que a Campanha Mundial de Erradicação da Febre Amarela da FR fortaleceu a cooperação interamericana em saúde, estreitando as relações entre as Repúblicas americanas entre as décadas de 1920 e 1940, processo que resultaria na Campanha para erradicação do Aedes aegypti que constitui-se em uma nova fase da campanha da FR, só que em um outro contexto internacional. Assim, através da análise da Campanha, com seus avanços, retrocessos e inflexões,em diferentes contextos políticos e sanitários, se discute a crescente cooperação internacional em saúde que vai se estabelecendo nas Américas ao longo do seu desenvolvimento. / This Ph.D. dissertation focuses on the Continental Campaign for the Eradication of Aedes aegypti, launched by the Pan American Health Organization (PAHO) in 1947. Its origins, however, date back to the Worldwide Campaign for the Eradication of Yellow Fever - conceived in 1914 by Wycliffe Rose, the first Director of the International Health Board (IHB) of the Rockefeller Foundation (RF) - and officially launched in 1918, after the end of World War I (1914-1918). The Campaign was developed between the 1910s and 1930s, in the Americas and Africa, and was marked by a series of inflections until its reformulation during World War II (1939- 1945) and was relaunched in 1947, under the auspices of PAHO, at that time directed by Fred L. Soper (1947-1959), a former official of the Rockefeller Foundation, with a long career in South America, where he fought diseases like hookworm, malaria and yellow fever. From this moment until the late 1960s, the goal of eradicating the vector of yellow fever of the Americas was pursued, with greater or lesser intensity, by practically all the American Republics. My aim is to analyze the origins, the historical development, and both the impacts and the controversies raised by this Campaign, which was the first and most lasting international eradication program already implemented. My hypothesis is that the Rockefeller’s Worldwide Campaign for the Eradication of Yellow Fever strengthened inter-American cooperation in health, narrowing the relationship between the American Republics between 1920 and 1940, a process which resulted, in the aftermath of World War II, in the Continental Campaign for the Eradication of Aedes aegypti which, as I tried to demonstrate, constituted a new phase of the RFcampaign, but in a new international context. Therefore, through the analysis of the Campaign, with its advances, setbacks and inflections, in different political and health contexts, I intend to discuss the growing international cooperation in health which was eventually settling in the Americas throughout its development.