Submitted by Alisson Mota (alisson.davidbeckam@gmail.com) on 2015-07-08T20:15:35Z No. of bitstreams: 1 Tese - Norma Suely de Lima Freitas.pdf: 8476174 bytes, checksum: b8e0fdb20a7b419aea33bc83321484fa (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-07-09T13:56:33Z (GMT) No. of bitstreams: 1 Tese - Norma Suely de Lima Freitas.pdf: 8476174 bytes, checksum: b8e0fdb20a7b419aea33bc83321484fa (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-07-09T13:54:31Z (GMT) No. of bitstreams: 1 Tese - Norma Suely de Lima Freitas.pdf: 8476174 bytes, checksum: b8e0fdb20a7b419aea33bc83321484fa (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-07-09T14:05:09Z (GMT) No. of bitstreams: 1 Tese - Norma Suely de Lima Freitas.pdf: 8476174 bytes, checksum: b8e0fdb20a7b419aea33bc83321484fa (MD5) / Made available in DSpace on 2015-07-09T14:05:09Z (GMT). No. of bitstreams: 1 Tese - Norma Suely de Lima Freitas.pdf: 8476174 bytes, checksum: b8e0fdb20a7b419aea33bc83321484fa (MD5) Previous issue date: 2012-08-28 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Despite the high prevalence and the risks associated with Chlamydia trachomatis in Brazil and other countries, little is known about the distribution of genotypes in Brazil and the biological variability of this important transmitting agent of sexually transmitted diseases (STDs). The absence of an inquiry routine for C. trachomatis and effective treatments can cause serious complications and consequences for individuals as pelvic inflammatory disease, infertility, ectopic pregnancy and neonatal infections. Currently, C. trachomatis presents 19 serotypes A-C associated with trachoma, D-K responsible for urogenital infections and L1, L2 and L3, agents responsible for lymphogranuloma venereum syndrome. The MOMP, which is recognized as the immunodominant antigen encoded by the ompA gene displays a large area of nucleotide variation of four variable domains (VDI to VDIV). Studies suggest that mutations occur frequently among genotypes and that these mutations may indicate differences between the immunogenic MOMP genotypes of C. trachomatis. The present work aims to identify the genotypes from samples positive by PCR for C. trachomatis in women diagnosed with infertility and with pregnant women in the city of Manaus, Amazonas - Brazil, in addition to sequence and analyze the ompA gene. The study population consisted of 96 infertile women and 53 pregnant women. Genotyping was performed by the technique of Polymerase Chain Reaction (PCR) from the ompA gene sequence and the following genotypes were identified in pregnant women: D [50.0%], E [25.0%] and F [12.5%] and I [12.5%]. In infertile women genotypes were identified: E [16.7%] F [16,7%] and K [66,7%]. The frequency of genotype K and D found in this study are considered high (66,7%) and (50,0%) and for phylogenetic analysis, we found that the genotypes analyzed shares the same ancestor. It is suggested that these variations in the sequences of genotypes identified arise from point mutations, or possibly by VD recombination in MOMP. The VDII region showed to be the most variable in the sequences analyzed. / Apesar da alta prevalência e dos riscos associados à Chlamydia trachomatis no Brasil e outros países do mundo, pouco se conhece sobre a distribuição dos genótipos no Brasil e a variabilidade biológica deste importante agente transmissor de doenças sexualmente transmissíveis (DST). A ausência de uma investigação rotineira para C. trachomatis e tratamentos efetivos pode originar sérias complicações e conseqüências para os indivíduos como doença inflamatória pélvica, infertilidade, gravidez ectópica e infecções neonatais. Atualmente, a C. trachomatis apresenta 19 sorotipos: A-C associados ao tracoma, D-K responsáveis por infecções urogenitais e L1, L2 e L3, agentes responsáveis pela síndrome do linfogranuloma venéreo. A MOMP, reconhecida como o antígeno imunodominante codificado pelo o gene ompA, exibe uma extensa área de variação nucleotídica sendo por sua vez, conferido por quatro domínios variáveis (VDI a VDIV). Estudos sugerem que as mutações ocorrem frequentemente entre os genótipos e que essas mutações podem indicar diferenças imunogênicas entre as MOMP de genótipos de C. trachomatis. O presente trabalho tem por objetivo identificar os genótipos a partir de amostras positivas por PCR para C. trachomatis de mulheres com diagnóstico de infertilidade e em gestantes na cidade de Manaus, Amazonas – Brasil, além de sequenciar e analisar o gene ompA. A população de estudo consistiu de 96 mulheres inférteis e 53 mulheres gestantes. A genotipagem foi feita pela a técnica de Reação em Cadeia de Polimerase (PCR) a partir da seqüência do gene ompA e os seguintes genótipos foram identificados em gestantes: D [50,0%]; E [25,0%]; F [12,5%] e I [12,5%]. Em mulheres inférteis os genótipos identificados foram: E [16,7%], F [16,7,%] e K [66,7%]. A freqüência de genótipo K e D encontrada neste estudo são consideradas elevadas (66,7%) e (50,0%) e quanto à análise filogenética, verificamos que os genótipos analisados compartilham do mesmo ancestral. Sugere-se que as variações encontradas nas sequências dos genótipos identificados surgem dos pontos de mutação ou possivelmente pela recombinação dos VD na MOMP. A região VDII foi que mais apresentou variações nas seqüências analisadas.

Protein aggregation is deleterious to human health and detrimental to therapeutic shelf-life. The physical processes that induce aggregation are the same processes that drive productive folding reactions. As such, protein aggregation is a non-productive form of protein folding. To gain insight into the steps that serve as a partition between the folding and aggregation reactions, the folding mechanisms of several β-rich proteins with links to human disease or medicine were examined. In the ALS-linked protein, SOD1, a subpopulation of the unfolded ensemble is found to be a common source of both nonnative structure and frustrated folding. These behaviors are only observed upon the reduction of the intrinsic disulfide bond, indicating that this covalent interaction wards against aggregation. The nonnative structure presents an attractive target for the development of new therapeutic agents. In VH domains from therapeutic mAbs, the intramolecular disulfide bond protects against aggregation. However, it can also introduce complexity to the folding mechanism. This complexity is linked to the formation of a strained orientation of the disulfide bond. This strained orientation of the disulfide in certain VH domains is energetically unfavorable enough to disrupt the formation of the disulfide in the full length mAbs. The novel relationship observed between disulfide orientation, folding complexity, and incomplete oxidation warrants further examination in other Ig domains. Overall, these results demonstrate that mapping the folding free energy landscape for proteins with roles in human disease or therapeutics can provide valuable insights for developing and improving treatment options.