F. tularensis is an intracellular pathogen, and is the causative agent of tularemia in humans. The ability of F. tularensis to parasitize host cells is largely dependent upon genes within a pathogenicity island (FPI), including those in the iglABCD operon. Specific mechanisms and gene products involved in regulation of the FPI are not well understood. I initiate the study of this regulatory system by creating an efficient Tn5-based mutagenesis system optimized for use in F. tularensis, and utilize this system to construct a lacZ reporter library. I identify genes differentially regulated in response to growth on two different media, including those in the iglABCD and fslABCD operons, and identify iron availability as a factor contributing to the differential regulation. One of these reporter strains, carrying a chromosomal iglB-lacZ fusion, is used as the basis for a secondary transposon mutagenesis to identify mutations that affect iglABCD expression. One such mutation is in FTL_1542 (migR), a hypothetical protein, and reduces expression of the iglABCD approximately 8-fold. The effect of this mutation on igl expression is likely through its effect on another known virulence regulator, fevR, as demonstrated by data from RT-PCR experiments. I compare the phenotypes of LVS fevR and migR mutant strains in primary macrophage and epithelial cell lines and in neutrophils. The mutation in migR effects growth and intracellular trafficking in macrophages but not epithelial cells, and reverses the ability of wild type F. tularensis to block the respiratory burst in neutrophils. When similar mutations were examined in the human virulent F. tularensis strain Schu S4, migR retained its regulatory role, but did not impair replication in macrophages. The migR mutation in Schu S4 did however have an attenuating effect when administered to mice intranasally. Comparison of LVS and Schu S4 in primary human airway epithelial cell infections revealed an inability of LVS to replicate within these cells, which is in contrast to the robust replication of LVS in cultured epithelial cell lines. Together, this work contributes to the understanding of regulatory mechanisms governing virulence gene expression in F. tularensis and highlights differences between LVS and Schu S4 strains.

Made available in DSpace on 2016-04-04T18:28:02Z (GMT). No. of bitstreams: 1 Vivian Mascella.pdf: 1282816 bytes, checksum: b8e395c9ccb4c074672d95f2c19c8223 (MD5) Previous issue date: 2011-12-02 / The current research sought to evaluate stress, anxiety and depression in women whith Migraine and women whith Tension-type Headache. A total of 31 women participated in the research .Sixteen out of 31 women suffered from Migraine and 15 out of 31 women showed Tension-type Headache. All the participants were between 18 to 72 years old. After being diagnosed by doctors, the participants were attended at a neurology and brain surgery clinic located in the interior of the State of S?o Paulo. A Semi-Structured Interview Form, Lipp Inventory of Stress Symptoms for Adults (LISS), Beck Anxiety Scale (BAS), and Beck Depression Scale (BDS) were used to collect data.The analysis of the data was both quantitative and qualitative. For LISS, BAS, and BDS, the answers were evaluated according to the charts and to the norms of their manuals. For the open answers of Interview form, the analysis of contend was made according to Bardin.The results revealed that 100% of the women who had Migraine showed stress; 50% of them were in a resistant period and 43,75% were in a nearly-exhausted period. 66,67% of the women who had Tension-type headache showed stress; 53,33% of them were in a resistant period and 6,67% in a nearly-exhausted period. Concerning anxiety, 31,25% of the women who had Migraine revealed a moderate level, while 60% of the women who had Tension-type Headache showed a minimum level. Concerning depression, 37,50% of the women who had Migraine showed moderate depression, and 53,33% of the women who had Tension-type Headache showed minimum levels of anxiety. It was observed that women who had Migraine showed more serious levels of stress, anxiety and depression than women who had Tension-type Headache. It is relevant to note that, although the survey sample is decreased, the results found are similar to other studies, and confirms the need to develop a psychological treatment suitable for women who suffer from Migraine and tension-type Headache, aiming to promote health and improves the quality of life. / A presente pesquisa teve como objetivo avaliar stress, ansiedade e depress?o em mulheres com Migr?nea e mulheres com Cefaleia do Tipo Tensional. As participantes foram 31 mulheres, sendo que 16 apresentavam Migr?nea e 15 apresentavam Cefaleia tipo tensional, na faixa et?ria de 18 a 72 anos. As participantes foram encaminhadas ap?s o diagn?stico dos m?dicos e atendidas numa cl?nica de Neurologia e Neurocirurgia no interior do Estado de S?o Paulo. Para a coleta dos dados, foi utilizado um roteiro de entrevista semiestruturada, Invent?rio de Sintomas de Stress para Adultos de Lipp (ISSL), Escala de Ansiedade de Beck (BAI), Escala de Depress?o de Beck (BDI). A an?lise dos dados foi quantitativa e qualitativa. As respostas dadas ao ISSL, BAI e BDI foram avaliadas de acordo com as tabelas e normas de seus manuais, e as respostas abertas foram submetidas ? an?lise de conte?do de Bardin. Os resultados revelaram que 100% das mulheres com Migr?nea apresentavam stress, sendo que 50% estavam em fase de resist?ncia e 43,75% se encontravam em fase de quase-exaust?o. Enquanto que 66,67% das mulheres com Cefaleia tipo tensional apresentavam stress, sendo que 53,33% estavam em fase de resist?ncia e 6,67% em fase de quase-exaust?o. No que se refere ? ansiedade 31,25% das mulheres com Migr?nea estavam num n?vel moderado, enquanto que 60% das mulheres com Cefaleia tipo tensional apresentavam um n?vel m?nimo. Com rela??o ? depress?o, 37,50% das mulheres com Migr?nea estavam em n?veis moderados, e 53,33% das mulheres com Cefaleia tipo tensional apresentavam n?veis m?nimos de ansiedade. Verifica-se que as mulheres com Migr?nea apresentam n?veis mais graves de stress, ansiedade e depress?o do que mulheres com Cefaleia tipo tensional. ? relevante destacar que, embora a amostra da pesquisa seja reduzida, os resultados encontrados s?o semelhantes aos de outros estudos, e confirma a necessidade de elabora??o de um tratamento psicol?gico adequado para mulheres que sofrem com Migr?nea e Cefaleia tipo tensional, visando ? promo??o da sa?de e melhora da qualidade de vida.

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