Introdução: O enfermeiro é responsável em atuar no controle da contaminação do ambiente, visando a prevenção de transmissão de infecções relacionadas à assistência à saúde. Surtos de gastroenterite causados por norovirus (NoV) em locais fechados são caracterizados pela persistência do vírus no ambiente, aerolização das partículas virais e baixa dose infectante, mesmo em indivíduos saudáveis. Portanto, há necessidade de definição de um protocolo seguro para limpeza e desinfecção do piso contaminado com vômito e fezes, considerando a possibilidade de dispersão de aerossóis a partir do piso. Objetivo: Avaliar a presença residual de partículas de NoV-GII no ar e no piso quando implementados diferentes protocolos de descontaminação do piso, após contaminação intencional. Método: Trata-se de um estudo experimental laboratorial. Dois tipos de piso, vinil e granito (matérias primas frequentemente utilizadas nos pisos dos serviços de saúde), foram contaminados intencionalmente com fezes humanas positivas 10% para NoV-GII, dissolvidas em 500ml de solução tampão salino-fosfato. Os pisos foram submetidos a três tipos de tratamento: limpeza padronizada com fricção manual, água e detergente neutro; limpeza seguida de desinfecção com hipoclorito de sódio 1% por 10 minutos; limpeza seguida de desinfecção com dispositivo portátil de luz ultravioleta por cinco minutos (SURFACE-UV®). Amostras foram obtidas do piso, por meio do swab, e do ar, por meio de um coletador de ar (Coriolis® - Bertin Technologies, França), nos seguintes momentos: antes e após a contaminação intencional; após a limpeza e após os métodos de desinfecção. Para detecção de NoV-GII nas amostras, utilizou-se a técnica 4.6.2. Reação em Cadeia pela Polimerase quantitativa precedida de Transcrição Reversa (RT-qPCR), pelo método TaqMan®. Resultados: Não foram encontradas diferenças estatisticamente significantes entre os tipos de piso após os protocolos de descontaminação, tanto para o residual de NoV-GII no piso, quanto no ar. Os valores médios de Cycle Threshold (Ct) após limpeza seguida de desinfecção foram maiores (38,75 40,00) comparados aos de após limpeza (35,67 38,66), comprovando a maior eficácia desse protocolo (p<0,001). Em algumas amostras, a limpeza isolada foi capaz de reduzir a contaminação por NoV do piso até níveis indetectáveis. Quando houve residual de NoV-GII após a limpeza do piso, o protocolo cuja desinfecção foi realizada com hipoclorito de sódio foi mais eficaz do que a luz ultravioleta (p<0,001), sendo que os valores de Ct de todas as amostras foram acima de 40. Em 27 das 36 (75%) amostras de ar coletas após a limpeza do piso, foram detectadas partículas de NoV, com diferenças estatisticamente significantes entre as segundas e terceiras amostras, coletadas a 150cm do piso. Foram identificadas que, em média, 17 cópias de RNA viral/L estavam presentes no ar após a limpeza, com redução gradual após a desinfecção. Conclusões: Quando vômito e fezes com NoV-GII contaminam o piso, há aerolização desse vírus já no ato da limpeza. Essas partículas podem ser inaladas ou depositarem em superfícies frequentemente tocadas pelas mãos, estabelecendo o ciclo de transmissão oro-fecal. As partículas virais residuais no piso após a limpeza, indiscutivelmente devem ser eliminadas, evitando assim a reaerolização do NoV a partir dessa fonte. Nesse sentido, a limpeza seguida de desinfecção com hipoclorito de sódio a 1% por 10 minutos mostra superioridade como protocolo de descontaminação do piso, quando comparado ao protocolo com limpeza seguida de desinfecção com luz ultravioleta por 5 minutos de exposição. / Introduction: Nurses are responsible for controlling contamination of the environment, working to prevent the transmission of health-care-associated infections. Gastroenteritis outbreaks caused by Norovirus (NoV) in closed settings are characterized as the result of persistence of the virus in the environment, aerosolization of viral particles, and small infectious dose, even in healthy individuals. Therefore, a safe protocol to decontaminate the floor after vomit or feces have spilled must be defined, considering that subsequent aerosol dispersal may occur. Objective: To assess the presence of residual NoV-GII particles in the air and on the floor after different decontamination protocols are conducted on a deliberately contaminated floor. Method: This is an experimental laboratory study. Two types of floor, vinyl and granite (materials which are often used in the flooring of healthcare facilities), were intentionally contaminated with 10% NoV-GII-positive human feces dissolved in 500ml of a saline-phosphate buffer solution. The floors received three types of treatment: standard cleaning, with manual friction, water, and neutral detergent; cleaning followed by a ten- minute disinfection procedure using 1% sodium hypochlorite; and cleaning followed by a five- minute disinfection procedure using a portable ultraviolet light device (SURFACE-UV®). Swab samples were taken from the floor, and air samples were obtained using an air sampler (Coriolis® - Bertin Technologies, France) at the following moments: before and after the intentional contamination, after cleaning, and after disinfection. The TaqMan® method for real-time Reverse Transcription-Polymerase Chain Reaction was used to detect NoV-GII in the samples. Results: No statistically significant difference between the two types of floor was found for residual NoV-GII, either in the air or on the floor, after the decontamination protocols. The average Cycle Threshold (Ct) values found after cleaning followed by disinfection were higher (38.75 - 40.00) than those recorded after cleaning (35.67 - 38.66), thus attesting to the greater effectiveness of the latter protocol (p<0.001). In some samples, cleaning alone was enough to reduce floor contamination by NoV to undetectable levels. When residual NoV-GII was found after cleaning the floor, the disinfection protocol that involved using sodium hypochlorite proved more effective than UV-light exposure (p<0.001), and Ct values were higher than 40 for all samples. NoV particles were detected in 27 of the 36 (75%) air samples obtained after cleaning the floor, and significant statistical differences were found between the second and third samples, collected 150cm from the floor. An average of 17 copies of viral RNA/L were identified in the air after cleaning, gradually decreasing after disinfection. Conclusions: When NoV-GII-infected vomit or feces contaminate the floor, the virus is aerosolized even during cleaning. These particles may then be inhaled or settle on frequently touched surfaces, establishing the fecal-oral transmission cycle. Residual viral particles on the floor must undoubtedly be eliminated, thereby preventing NoV aerosolization from this source. Along these lines, cleaning followed by disinfection by 1% sodium hypochlorite for ten minutes proved to be a superior floor decontamination protocol when compared with cleaning followed by disinfection by UV-light exposure for five minutes.

Introdução: O crescente aumento do número de casos de AIDS em mulheres evidencia um progressivo aumento da Transmissão Vertical (TV) do HIV, gerando assim a necessidade de focalizarmos esforços no intuito de impedir ou minimizar este tipo de transmissão. Objetivos: avaliar as condutas profiláticas adotadas para redução da TV do HIV em serviços de referência pertencentes à 7ª CRS. Metodologia: Os dados foram obtidos por meio de pesquisa de campo retrospectiva com análise quanti-qualitativa dos formulários de cadastro, fichas de notificação e investigação compulsória, além de prontuários das gestantes e puérperas soro positivas para o HIV, no período de 01 de janeiro de 2010 a 01 de janeiro de 2013 mediante aprovação ética. Resultados: A população estudada compreendeu um universo de 22 mulheres notificadas como Gestante HIV +, dentre as quais 72% foram notificadas na cidade de Bagé, e destas, 9% foram notificadas em 2010, 27,3% em 2011 e 63,7% em 2012. A faixa etária de maior prevalência é de 15 a 24 anos (59%), 50% das mulheres não teve a via de parto nem a evolução da gravidez informada e 60% não tem registro de profilaxia no período intra-parto. Em 50% das fichas, não foi informado o uso da medicação pela criança nas primeiras 24h e 59% das notificações foram feitos por profissional enfermeiro. Pôde-se caracterizar com os dados coletados os aspectos individual, escolar e sócio-demográfico da 7ª CRS. Indo ao encontro das informações publicadas por Brasil (2013), este estudo evidencia uma sub-enumeração de casos no SINAN, pois apenas 63% das mulheres foram notificadas/cadastradas nos sistemas de informação. Considerações: As práticas de saúde adotadas pelos serviços estão de acordo Protocolo Vigente, no que se trata da administração da TARV a partir da 14ª semana de gestação, da escolha, sempre que possível da via de parto, a administração de medicamentos intra-parto, a inibição da lactação, o uso do ARV no bebê e o fornecimento de fórmula láctea. As medidas educativas necessárias para qualificar o atendimento realizado na região de saúde estudada relacionam-se principalmente à qualidade das informações das ações de saúde (alimentação dos bancos de dados dos sistemas), além de estimular a população em geral para o conhecimento do prénatal e das testagens para HIV. Para que pudéssemos trabalhar acerca deste tema, discutimos os resultados do estudo em equipe no SAIS de Bagé, e pensamos em algumas estratégias que nos aproxime dos demais profissionais da rede. Criamos um perfil em uma rede social, onde estão sendo postadas diariamente informações acerca dos temas: HIV, AIDS, DSTs, Hepatites Virais, medidas de prevenção, meios de transmissão, atividades do serviço, dados importantes, dentre tantas outras informações, sendo imprescindível relatar que todos os componentes da equipe tem acesso a senha, para que possa sentir-se ator no processo de educação da comunidade. Logo, serão propostas oficinas e fóruns, com os temas em questão para discutir com a comunidade local, com o apoio da equipe da 7ª CRS/RS. / The increasing number of AIDS cases among women reveals a progressive increase of Vertical Transmission (VT) of HIV, thus generating the need of focusing efforts in order to prevent or minimize this type of transmission. The aim of the study was to evaluate the prophylactic measures adopted to reduce HIV in TV reference belonging to the 7th CRS services. Data were obtained from retrospective research field with quantitative and qualitative analysis of the registration forms, reporting forms and compulsory research, and medical records of pregnant and postpartum serum positive for HIV in the period from January 1, 2010 to January 1, 2013 by ethical approval. The study population comprised a universe of 22 women reported / registered as HIV + Pregnant Women, among which 72% were reported in the city of Bage, while 09% reported in 2010, 27.3% in 2011 and 63.7% in the year 2012 the most prevalent age group is 15-24 years (59%), 50% of women had no way of delivery, or pregnancy outcome informed and 60% have no record of prophylaxis in intra period partum. In 50% of the chips, was not informed use of medication by the child in the first 24h and 59% of registrations / notifications were made by professional nurses. Could be characterized with the data the individual, school and socio-demographic aspects of CRS 7th. Going against the information published by Brazil (2013), this study highlights an under-enumeration of cases in SINAN because only 75% of women were reported / registered in information systems. Practices adopted by health services meet the Effective Protocol, it is the administration of ART from the 14th week of gestation, the choice where possible the mode of delivery, medication administration intra-partum, the inhibition of lactation, the use of ARVs in providing baby and milk formula. Educational measures to improve service held at the health region studied relate primarily to information quality of health (nutrition database systems), in addition to encouraging the general population to the knowledge of prenatal and testings for HIV. So we could work on this subject, discuss the results of the study team at SAIS Bage, and think of some strategies that bring us closer to the other networking professionals. We created a profile on a social network, where they are being posted daily information on the topics: HIV, AIDS, STDs, Viral Hepatitis, prevention, transmission media, service activities, important data, among many other information, is indispensable report that all team members have access to the password, so you can feel an actor in the process of educating the community. Soon, workshops and forums will be proposed, with the issues in question to discuss with the local community, with the support of the 7th CRS / RS staff.

Submitted by (edna.saturno@ufrpe.br) on 2016-10-21T12:41:27Z No. of bitstreams: 1 Pedro Alves de Moura Sobrinho.pdf: 655310 bytes, checksum: cf5f69ddf5c19bb603c77fd7a7012e4e (MD5) / Made available in DSpace on 2016-10-21T12:41:27Z (GMT). No. of bitstreams: 1 Pedro Alves de Moura Sobrinho.pdf: 655310 bytes, checksum: cf5f69ddf5c19bb603c77fd7a7012e4e (MD5) Previous issue date: 2008-02-04 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The aim of the present study was to characterize the goat and sheep production systems, estimate the prevalence of small-ruminant lentiviruses (SRLV) and study predisposing factors to infection in herds in the state of Tocantins (Brazil). Investigative questionnaires were administered addressing data on 29 goat herds and 28 sheep herds distributed among 21 towns in the state of Tocantins. To estimate the prevalence of SRLV, 838 samples of sheep blood and 843 samples of goat blood were analyzed using the immunodiffusion test in agarose gel. There was a mean number of 79 and 340 animals for the goat and sheep herds, respectively. According to the survey, 79.3 and 71.4% of the goat and sheep farms, respectively, were implanted after 2001. The animals making up the herds originated in the states of Bahia and Sergipe. The Santa Inês (61.0%) and SRD (38.7%) were the most frequent breeds among the sheep, whereas the Anglo-Nubian (55.4%), SRD (36.6%) and Saanen (7.1%) were the most frequent goat breeds. 55.1% of the 49 properties surveyed bred cattle + goats or sheep. The semi-extensive system was adopted in 51.7 and 46.4% of the goat and sheep farms, respectively. The most frequently adopted hygiene practices among the breeders were de-worming, cuts and disinfection of the navel and burying dead animals. Vaccination is practiced by 24.1% of goat breeders, most frequently for clostridiosis (20.7%), aphthous fever (6.8%) and rabies (3.4%). The clinical signs and diseases most often cited as having considerable importance were caseous lymphadenitis (34.5%) and pododermatitis (17%) by goat breeders and pododermatitis (42.3%) and diarrhea (30.8%) by sheep breeders. 51.7 and 28.6% of the goat and sheep breeders, respectively, knew about lentiviruses (CAE and Maedi-Visna). The frequency of ovine blood reagents was 0.9 ±0.3% (8/838). The results regarding the micro-regions of the state were distributed in the following manner: 3.3 (2/60), 0.61 (1/178), 1.3 (2/150) and 2.0% (3/150) for Bico Papagaio, North, Miracema and South, respectively. No animals tested positive in the Jalapão, Porto Nacional, Rio Formoso and Southeast micro-regions. The Santa Inês was the breed with the highest percentage of blood reagent animals (3.9%; 6/511), followed by Undefined Breed (0.6%; 2/324). Regarding age, 0.6% (2/238) and 1.2% (96/510)of animals under 24 and over months of age, respectively, tested positive. 1.2% (2/161) of males and 0.9% (6/677) of females tested positive. There were no statistically significant differences in the prevalence of blood reagent regarding breed,age and gender (p > 0.05). The frequency of blood reagent goats was 2.7% (23/843). The results were distributed according to micro-region in the following manner: 10.0 (20/200) and 1.4% (3/207) for the North and Miracema micro-regions, respectively. The Saanen was the breed with the highest percentage of blood reagent animals (11.7%; 7/60), followed by the SRD (0.6%; 2/310) and Anglo-Nubian (3.0%; 14/466). Regarding age, 1.9% (6/314) and 3.2% (17/529) of animals under 24 and over months of age, respectively, tested positive. 2.4% (4/166) of males and 2.8% (19/677) of females tested positive. Analysis of the data revealed a predominance of the semiextensive breeding system with good installations, but little use of reproduction biotechniques and important hygiene management practices. The activity has been expanding in the state, with a tendency toward professionalization. SRLV infection occurs at a low prevalence among sheep and goats in the state of Tocantins and is evenly distributed according to micro-region, breeding system, breed, age and origin of the base herds. Control measures should be implanted to avoid the dissemination of the disease among the herds. / Objetivou-se com esta pesquisa caracterizar o sistema de produção de caprino e ovino, estimar a prevalência e estudar os fatores predisponentes à infecção por Lentiviroses de Pequenos Ruminantes (LVPR), em rebanhos no Estado do Tocantins. Foram aplicados questionários investigativos em 29 rebanhos de caprinos e 28 de ovinos, distribuídos em 21 municípios do Estado do Tocantins, sendo 15 para cada espécie. Para estimar a prevalência de LVPR, foram analisadas 838 amostras de soros ovinos e 843 amostras de caprinos, utilizando o teste de imunodifusão em gel de agarose – IDGA. Identificou-se um número médio de animais de 79 e 340 para os rebanhos de caprinos e ovinos, respectivamente. Conforme a pesquisa, 79,3 e 71,4% dos criatórios de caprinos e ovinos, respectivamente, foram implantados após 2001. Os animais para formação dos rebanhos bases tem como origem os Estados da Bahia e Sergipe. Os tipos raciais Santa Inês (61,0%) e o SRD (38,7%) são os mais encontrados entre os ovinos e na espécie caprina o Anglo-nubiano (55,4%), SRD (36,6%) e Saanen (7,1%). O estudo mostrou que 55,1% das 49 propriedades pesquisadas criam bovinos + caprinos ou ovinos. O sistema semi-extensivo é adotado em 51,7 e 46,4% dos criatórios de caprinos e ovinos, respectivamente. As práticas sanitárias adotadas com maior freqüência pelos criadores são vermifugação, cortes e desinfecção do umbigo e enterrar os animais mortos. A vacinação é prática adotada por 24,1% dos criadores de caprinos, sendo as mais freqüentes contra clostridioses (20,7%), febre aftosa (6,8%) e raiva (3,4%). Os sinais clínicos e doenças mais citados como de grande importância foram linfadenite caseosa (34,5%) e pododermatite (17%) pelos criadores de caprinos e Pododermatite (42,3%) e diarréias (30,8%) para os ovinos. Dos produtores de caprinos e ovinos, 51,7 e 28,6%, respectivamente, conhecem as lentiviroses (CAE e a Maedi-Visna). A frequência de ovinos sororreagentes encontrada foi de 0,9 ±0,3% (8/838). De acordo com a microrregião do Estado, os resultados foram assim distribuídos: 3,3 (2/60), 0,61 (1/178), 1,3 (2/150) e 2,0 % (3/150) para Bico Papagaio, Norte, Miracema e Sul e Sudeste, respectivamente. As microrregiões Jalapão, Porto Nacional, Rio Formoso e Sudeste não tiveram animais positivos. Entre as raças, a Santa Inês foi a que apresentou numericamente o maior percentual de animais sororreagentes, 3,9% (6/511), seguida da Sem Raça Definida, 0,6% (2/324). De acordo com a idade, os animais com idade inferior e superior a 24 meses apresentaram 0,6% (2/328) 1,2% (6/510), respectivamente. Os machos apresentaram 1,2% (2/161) de positivos e as fêmeas 0,9% (6/677). Não houve diferença significativa (p > 0,05) na prevalência de sororreagentes com relação às variáveis raça, idade e sexo. A frequência de caprinos sororreagentes encontrada foi de 2,7% (23/843). De acordo com a região, os resultados foram assim distribuídos: 10,0 (20/200) e 1,4% (3/207) para as microrregiões Norte e Miracema, respectivamente. Entre as raças, a Saanen foi a que apresentou numericamente o maior percentual de animais sororreagentes, 11,7% (7/60), a SRD 0,6% (2/310) e a anglonubiana 3,0% (14/466). De acordo com a idade, os animais com idade inferior e superior a 24 meses apresentaram 1,9% (6/314) e 3,2% (17/529)), respectivamente. Os machos apresentaram 2,4% (4/166) de positivos e as fêmeas 2,8% (19/677). A análise das informações mostrou uma atividade onde predomina o sistema de criação semiextensivo com boas instalações, mas pouco uso de biotécnicas da reprodução e uso de importantes práticas de manejo sanitário. Observou-se que a atividade vem se expandindo no Estado com uma tendência de profissionalização. A infecção por LVPR ocorre em ovinos e caprinos do Estado de Tocantins com baixa prevalência, homogeneamente distribuída de acordo com a microrregião, sistema de criação, raça, sexo, idade e origem dos rebanhos base. Medidas de controle devem ser implantadas no sentido de evitar a disseminação da doença entre os rebanhos.

Introdução: A tuberculose é uma doença de repercussão mundial que desafia os serviços de saúde (SS). Estima-se que um terço da população mundial apresenta a forma latente da doença. Os profissionais da saúde (PAS) inseridos na assistência direta podem ter maior probabilidade de contágio; entretanto, os índices de ocorrência da doença nesta população não foram, até o momento, reconhecidos de forma sistemática. Objetivo: Estimar a prevalência e a incidência de tuberculose ocupacional nos serviços de saúde. Método: Trata-se de uma revisão sistemática de literatura segundo protocolo do Joanna Briggs Institute, que buscou responder quais os índices de tuberculose ocupacional nos SS. Foram considerados estudos primários que incluíssem dados de incidência e prevalência de tuberculose ocupacional ou de viragem tuberculínica em PAS. Foram definidos como PAS indivíduos em formação ou formados em cursos na área da saúde e que atuam na assistência direta. Quanto a SS, foram consideradas instituições de atendimento em qualquer um dos níveis assistenciais e que realizam atendimento direto a pacientes suspeitos ou portadores de tuberculose em suas fases transmissíveis, de forma a manter a homogeneidade da investigação. Foram excluídos os relatos de surtos e as ações dirigidas a populações em geral. Resultados: Foram identificadas 2.081 citações potencialmente relevantes em dez bases de dados, sem limites de tempo e idioma, a seleção final levou a 17 estudos. Em todos os artigos selecionados o desfecho estudado foi tuberculose latente. Não foram identificados estudos com dados com relação à tuberculose doença. O teste tuberculínico foi utilizado em 88,2% dos estudos. A prevalência de tuberculose latente (TL) variou entre 0,1% e 59,7% e a taxa de incidência anual variou de 3,3% a 26%. Um estudo identificou a densidade de incidência de 547/100.000 pessoas/ano. Conclusão: As evidências demonstram que a tuberculose é um importante agravo entre os PAS nos Serviços de Saúde, com importantes variações de acordo com o contexto. Os resultados indicam que a despeito da relevância do fenômeno, ainda há lacunas de informação sobre a incidência e a prevalência da tuberculose doença em PAS. / Introduction: Tuberculosis is a disease of worldwide repercussion that defies health care services (HS). It is estimated that one-third of the world population has a latent form of the disease. Healthcare workers (HCW) dealing with direct patient assistance may have higher contagion probability. However, the rates of the disease occurrence in this population have not been established in a systematic way yet. Objective: To estimate the global prevalence and incidence of occupational tuberculosis in HS by means of the literature review. Method: This is a systematic literature review according to the Joanna Briggs Institute, which sought to identify the rates of occupational tuberculosis in HCW. Primary studies were considered to include data on the incidence and prevalence of occupational tuberculosis or tuberculin skin test reaction in HCW. Reports of outbreaks and actions addressed to the community in general were excluded. Results: Out of ten cross-referenced databases, 2,081 potentially relevant studies were identified, without time and language limits, and the final selection was 17 studies. In all the articles selected, the outcome was latent tuberculosis. No studies with data regarding the tuberculosis disease were identified. The tuberculin skin test was used in 88.2% of the studies. The prevalence of latent tuberculosis ranged from 0.1 % to 59.7%. The annual incidence rate varied from 3.3% to 26%. As the risk and occupational exposure the absence of vaccination prior, the duration of care and performance in tuberculosis specific services were shown as relevant factors. One study identified the density of incidence of 547/100,000 people per year. Conclusion: Evidence indicates that tuberculosis is a relevant disease with the HCW and has important rates variations according to the context. The results demonstrate that despite the relevance of the phenomenon, there are still gaps in information on the incidence and prevalence of the tuberculosis disease in HCW.

O presente trabalho visa, através de métodos genético-epidemiológicos, identificarem genes associados à resposta ao antígeno da Leishmania. Utilizando amostras através do teste de Montenegro dos municípios de Monte Negro (RO) e Assis Brasil (AC). Na primeira abordagem foram feitos testes com TaqMan&#210 e a segunda com GWAS, e análises de associação foram feitas utilizando-se o pacote SPSS e o Plink. Não foram encontradas associações com cinco SNPs (MYD88, IL12, IL10, IFNGR1 e NRAMP1). A análise de dados de varredura genômica com filtros, indicou uma região no cromossomo 10 com 3 SNPs próximos que fazem parte de uma região regulatória que com o posterior auxilio do real time não se confirmaram, apesar do ensaio RS11251056 apresentar valores limítrofes, se tornando uma possível indicação para trabalhos futuros e por fim a último teste foi a meta-análise, através do método de Woolf, apresentou resultados indicativos de associação para ensaios encontrados no cromossomo 2 com ZNF638 relacionados a diferenciação celular e também no cromossomo 10 que contem lincRNAs e o gene NGR3, com dois ensaios apresentando valores significativos de p, onde podemos inferir que estas duas regiões podem participar ativamente na diferenciação da resposta ao antígeno da Leishmania. / The present study aims, through genetic-epidemiological methods, to identify genes associated with response to Leishmania antigen. Using samples Montenegro skin test through the municipalities of Monte Negro (RO) and Assis Brazil (AC). In the first approach were tested with TaqMan&#210 and the second GWAS, and association analyzes were performed using SPSS and Plink. No associations were found with five SNPs (MyD88, IL12, IL10, IFNGR1, and NRAMP1). The analysis of genome scan data with filters, indicated a region on chromosome 10 with three nearby SNPs that are part of a regulatory region that later with the help of real time is not confirmed, although the test rs11251056 have borderline values, becoming an possible direction for future work and finally the last test was the meta-analysis by the method of Woolf, presented results indicating the association found in tests for chromosome 2 with ZNF638 related to cell differentiation and also on chromosome 10 that contains lincRNAs and gene NGR3, two runs with a significant p value, where we can infer that these two regions can actively participate in the differentiation of the response to Leishmania antigen.

Histone deacetylases (HDACs) are key mediators of gene expression and, thus, major regulators of cell function. As such, HDACs play a role in orchestrating tumor biology, and the use of small inhibitors targeting theses proteins is attractive for the field of cancer therapy. Indeed, several HDAC inhibitors have received FDA-approval for the treatment of malignancies, while a myriad of these compounds continue to be evaluated in clinical trials. Besides their direct impact on tumor growth, HDAC inhibitors have been shown to increase immunogenicity of cancer cells, facilitating generation of a productive immune response against tumors. Immunotherapeutic approaches take advantage of the intrinsic ability of the immune system to manifest an anti-tumor response. Mechanisms of immune escape are often developed by cancer cells, neutralizing activity of the immune system. For example, upregulation of the PD1 ligands PDL1 and PDL2 by tumor cells negatively regulates the anti-tumor functions of PD1-expressing infiltrating T-cells. Importantly, strategies targeting this inhibitory axis have shown outstanding clinical benefit for the treatment of solid and hematological malignancies. The mechanisms by which HDAC inhibitors modulate tumor and immune cells biology were explored herein. Initially, treatment of melanoma cells with pan- and class I-selective HDAC inhibitors resulted in upregulation of PDL1 and PDL2 molecules. These effects were observed in mouse and human cell lines, as well as in tumor cells resected from metastatic melanoma patients. This upregulation was robust and sustained, lasting at least 96 hours in vitro, and validated in vivo using a B16F10 syngeneic mouse model. Enhanced expression of PDL1 mediated by HDAC inhibitors was found to result from enhanced histone acetylation at the PDL1 gene promoter region. Combination therapy of HDAC inhibition and PD1 blockade was explored in the tumor setting, leading to synergistic effects in terms of reducing melanoma progression and increasing survival of B16F10 melanoma-bearing mice. These data provide a clinical rationale for combination therapy of epigenetic modifiers (e.g. HDAC inhibitors) and PD1 blockade as means to augment cancer immunotherapy, improving patient outcomes. As a second pillar of this research, the impacts of HDAC-selective inhibition were explored on immune cell biology, since the broad nature of pan-HDAC inhibitors was shown to be detrimental to T-cells in vitro and in vivo. Based on screening assay results, novel implications of treating melanoma patient T-cells ex vivo with the HDAC6-selective inhibitor ACY1215 were investigated. Treatment with this compound was unique among pan- and isotype-selective HDAC inhibitors in modulating T-cell cytokine production and showing minimal impact of T-cell viability. ACY1215 tempered Th2 cytokine production (i.e. IL-4, IL-6 and IL-10), and maintained Th1 effector cytokines (e.g. IFNγ and IL-2). Furthermore, ACY1215 increased expression of surface markers, including CD69 activation marker and ICOS co-stimulatory molecule. In addition, ACY1215 treatment enhanced accumulation of central memory T-cells during ex vivo expansion of tumor infiltrating T-cells harvested from resected tumors of metastatic melanoma patients. Importantly, ACY1215-mediated inhibition improved tumor-killing capacity of T-cells. These results highlight an unexplored ability of selective HDAC inhibitor ACY1215 to augment T-cell expansion during protocols of adoptive cell therapy. While the discoveries presented here warrant further investigation of cellular and molecular mechanisms associated with ACY1215-treated T-cells, the clinic implications are clear and rapidly translatable.

Acute Lung injury (ALI) and the more severe acute respiratory distress syndrome (ARDS) are respiratory maladies that present immense clinical challenges. ALI affects 200,000 individuals annually and features a 40% mortality rate. ALI can be initiated by both pathogenic and sterile insults originating locally in the lungs or systemically. While immense research has been poured into this disease in an effort to find a therapeutic strategy, the heterogeneously diffuse nature of the disease has not yielded a cure for the disease. Death from this disease is strongly attributed to reduced gas exchange from a severely compromised alveolar-capillary barrier. The only way currently to manage this disease is through enhanced ventilation and hyperoxic therapy. Hyperoxic therapy is a common treatment given to over 800,000 patients each year to treat respiratory maladies such as ALI. Prolonged exposure to oxygen at high concentrations results in the development of a condition known as hyperoxic acute lung injury (HALI). In this disease, the formation of reactive oxygen species damages healthy tissue and impairs gas exchange. Hyperoxia is also a well-documented murine sterile lung injury model that replicates the symptoms of ALI in lung injury patients. The ability of non-lethal dosages of hyperoxia to resolve without lung fibrosis also enables the study of molecules associated with ALI resolution and repair, a process not clearly understood. Inflammation in ALI is associated with disease progression, however pharmaceutical interventions aimed at targeting the inflammatory cascade have failed in clinical trials for ALI. Recent reports point to an aberrant injury resolution mechanisms that may be more strongly correlated with morbidity and mortality. There seems to be a homeostatic imbalance between endogenous inflammation progression and resolution initiation. This is especially the case with HALI, as significant ROS generation results in depletion of redox regulating antioxidants. Resolution mechanisms associated with ALI in the oxygen toxicity setting is poorly understood. Polyunsaturated fatty acids such eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are essential fatty acids that show immense antioxidant and anti-inflammatory action in cases of acute injury. The lung mucosa is rich in DHA and following inflammatory insult DHA is readily converted to resolution phase interaction products (resolvins), which have shown immense proresolutionary potential in recent reports of acute injury. In the presence of aspirin, more potent and longer-acting aspirin-triggered resolvins are formed. The effects of resolvins and their aspirin triggered epimers have not been studied in an oxygen toxicity setting and are the focus of this dissertation. For the first time, we show that one of these resolvin molecules, aspirin triggered resolvin d1 (AT-RvD1), can enhance resolution of hyperoxic acute lung injury. In vitro results reveals that AT-RvD1 treatment resulted in reduced interaction of two key players in the HALI inflammatory cascade, the macrophage and alveolar epithelium. AT-RvD1 was able to blunt macrophage cytokine secretion as well as inhibit epithelial cell cytokine secretion and adhesion molecule expression. More importantly, AT-RvD1 blunted cytokine mediated leukocyte-epithelial cell interaction in vitro. In a sublethal hyperoxic injury model, mice given AT-RvD1 following hyperoxia exposure displayed reduced HALI pathological severity. ATRvD1 treatment resulted in reduced alveolar-capillary permeability, tissue inflammation, proinflammatory mediator secretion, epithelial cell death, and leukocyte influx. Taken together these novel results demonstrate the therapeutic potential of resolvins in the oxygen toxicity setting. These results also arouse the idea that resolvins could be used to lessen the comorbidities associated with oxygen therapy and improve recovery times of ALI patients.

Infections with dengue viruses (DENV) constitute both a global problem as well as locally in Florida. DENV comprise four distinct serotypes of single-stranded RNA viruses and belong to the family Flaviviridae. DENV are among the most medically important arboviruses in the world and cases may currently exceed 400 million per annum. Additionally, dengue established its first recorded endemic transmission cycle in the state of Florida in over a half century, first within the Florida Keys during 2009-10 followed by an unrelated outbreak in Martin County in 2013. The clinical profile of DENV infections ranges from a mild febrile illness to severe illness including hemorrhaging and/or shock, occasionally leading to death. Asymptomatic and mild cases also play a role in maintaining transmission cycles. The early diagnosis and management of patients at the clinical level have both proven to be major obstacles in the control of DENV and are important at both the individual and community levels. Individually, the proper management of patients that will progress to severe illness demands that they are identified in order to receive supportive treatment and mitigate associated morbidity and mortality. At the community level, early diagnosis may reduce transmission by limiting the possibility of vector contact with viremic individuals. Early diagnosis is dependent on the detection of viral markers, while a number of host factors may inform prognosis. The microsphere-based immunoassay (MIA) is capable of detecting up to 100 different targets in a single sample and therefore would be useful as a single assay for determining both. This study attempted to develop a diagnostic and prognostic MIA using the DENV NS1 glycoprotein and 5 host markers as targets. For the purposes of DENV NS1 detection in MIA, a set of monoclonal antibodies (mAbs) were subjected to immunoprecipitation and/or Western blot analysis in order to determine immunoreactivity. Two mAbs, 3A5.4 and 3D1.4, were chosen for use in MIA as a capture antibody and a detection antibody, respectively, and the results compared to a commercially available DENV NS1 ELISA. The 5 markers chosen for MIA trials included GM-CSF, IFN-γ, IP-10, IL-10, and MCP-1 and were selected from a panel of 27 markers screened initially in two in vitro models of DENV infection in addition to serum samples. The two cell lines investigated were HPMEC ST1.6R and u937 as both are thought to play important roles in models of DENV pathogenesis. The results of the DENV NS1 detection MIA were initially promising but were ultimately unsuccessful. When measuring host markers in the MIA, results pointed towards certain profiles that may be of future use. IL-10 was found to be elevated in a statistically significant manner in DENV qRT-PCR+ samples (p=0.035) when compared to negative sera. MCP-1 elevation was found to be of borderline significance (p=0.058). Other potential markers based on the results reported here include IP-10, IL-6, IL-8, VEGF, and RANTES. The ultimate goal of measuring host markers is to gain the ability to differentiate patients that will progress to severe illness from those that will recover. In conclusion, despite the failure of the MIA to detect DENV NS1 in human sera, our results in determining host markers and developments leading to successful DENV NS1 detection ELISAs elsewhere lead us to believe that this approach remains promising. Major drawbacks of this study included the lack of samples from patients that experienced severe DENV illness as a comparative group in addition to small sample sizes. Future goals should include determining the reasons for the failure of the MIA in detecting DENV NS1, selecting a panel of appropriate markers to differentiate non-severe from severe cases of DENV prior to progression, and optimizing the inclusion of these markers to an appropriate number.

Current predictions of pollen levels rely strictly on historical Averages, regardless of environmental factors that might affect the timing of pollen release by different plants. For this thesis, the goal was to develop a statistical model that will accurately forecast pollen levels by correlating those daily counts to atmospheric and meteorological conditions. This project used ARIMA modeling on IBM’s SPSS Statistics 24 of daily pollen count information for multiple allergenic pollens in the Sarasota County, Florida area over a 11-year period. The pollen species in question for this project are oak and cypress trees, grass, and ragweed pollens; and Alternaria and Cladosporium mold spores. The total pollen counts for weeds, grass, trees, and overall total are also included in the 11 years of data. The atmospheric variables used to predict pollen levels are high temperature, low temperature, average temperature, precipitation, humidity, wind direction, and wind speed for daily observations over the 11-year period. Results for these models showed that maximum temperature, precipitation, humidity, and wind direction were the driving predictors behind the pollen counts in Sarasota, Florida. The analysis of the pollination periods also showed that there were phenological changes according to the specific species. The models and phenological changes are specific to the Sarasota, Florida area, and would serve as a framework for studying other pollination regions.

Mast cells are critical effectors of allergic disease that can be activated by numerous stimuli. We have examined mast cell control by the inflammatory cytokine, IL-33, as well as IgG. In the first study reported here, we found that the synthetic glucocorticoid, dexamethasone, potently and rapidly suppressed IL-33-induced cytokine production from murine bone marrow–derived and peritoneal mast cells, as well as human mast cells. Dexamethasone also antagonized IL-33-mediated enhancement of IgE-induced cytokine production and migration. Although dexamethasone had no effect on IL-33-induced phosphorylation of MAP kinases or NFκB p65 subunit, it antagonized AP-1 and NFκB-mediated transcriptional activity. Finally, intraperitoneal administration of dexamethasone completely abrogated IL-33-mediated peritoneal neutrophil recruitment and prevented plasma IL-6 elevation. These data demonstrate that steroid therapy may be an effective means of antagonizing the effects of IL-33 on mast cells in vitro and in vivo, acting partly by suppressing IL-33-induced NFκB and AP-1 activity. In the second study reported here, we found that Fcγ receptor crosslinkage activated the transcription factor Stat5B through a Fyn kinase-dependent pathway. We then showed that STAT5B is critical for IgG-induced cytokine production by mast cells but not macrophages. To expand these studies, we employed the K/BxN model of inflammatory arthritis, which has roles for mast cells and macrophages. In this model, Fyn or STAT5B deficiency only affected the arthritic flare that primarily depends on mast cell degranulation, without affecting the severity of the joint swelling. By contrast, Lyn kinase deficiency significantly exacerbated arthritis. These studies indicate a clinically relevant, lineage-restricted role for the Lyn/Fyn-STAT5 cascade. Collectively, our work demonstrates that mast cell activation by diverse stimuli can be suppressed by steroid intervention and selectively targeted by disrupting kinase-transcription factor pathways.