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No. of bitstreams: 3 TESE_Évelyn Costa Lira.pdf: 5199119 bytes, checksum: 3542649866fd36a1d250872b70b7f22e (MD5) Diploma de Doutorado ou Ata de Defesa.pdf: 129558 bytes, checksum: 3cdafbe7ae427bf6a30e4dd767a402e5 (MD5) Carta de Encaminhamento para Autodepósito.pdf: 33896 bytes, checksum: cbb2fa1f7645982868cec33dd021fd5d (MD5) Previous issue date: 2017-11-30 / FAPEAM - Fundação de Amparo à Pesquisa do Estado do Amazonas / The role of the human microbiota in health and disease processes has received increased attention due to the ease of characterization of microbial communities using independent culture methods, which are based on the analysis of hypervariable regions of the gene 16S rRNA. The vaginal microbiota is inhabited by microbial communities that play a very important role in the maintenance of vaginal homeostasis and in the presence of colonization by pathogenic microorganisms, but the mechanisms by which they exert this influence are not yet so well defined. in this context, an understanding of their relative abundance and variations is necessary for the recognition of potential pathogenic microorganisms and the physiological processes of protection of this microbiota. This study evaluated the vaginal microbial diversity in four different conditions: I Microbiota Normal II Vaginal Candidosis Microbiota III Bacterial vaginosis microbiota IV microbiota presenting pre-malignant and malignant lesions of the cervix. Cervical samples from 187 women were collected, characterized and diagnosed at the molecular level for the presence of HPV, C. trachomatis, T. vaginalis e N. gonorrhoeae. HPV positive samples were genotyped at ABI 3500 and HPV co-infection were related to the presence of other pathogens and socio-economic and clinical factors obtained through the questionnaire filled out by the volunteers. Significance and association analyzes were performed using the Chi-square Test of Pearson, exact of Fischer’s Test and Kruskal-Wallis test using R 2.9.0. After the groups were composed, four libraries of amplicons of the regions V1-V2 of the gene 16S rRNA and, later sequenced on Ion PGM platform. The sequences generated were analyzed using the QIIME and classified by comparison in the Greengenes database. the prevalence found for HPV was 51.33%, and the most prevalent types were HPV 16, 58 and 33. The prevalence found for CT and TV were 6.42% and 12.83%, respectively, with co- HPV / CT infection in 5.20% and HPV / VT in 6.25% of the women. The DNA of N. gonorrhoeae were not found in samples. as to the estimation of microbial diversity, the number of samples sequenced was enough to guarantee coverage of the total diversity found. Overall abundancy revealed a predominance of the genus lactobacillus on the four study groups, followed by the genres Ureaplasma, Prevotella and Shuttleworthia, followed by the study groups. the greatest microbial diversity was found in the vaginosis group, with the most abundant genera Prevotella, Shuttleworthia and Megasphaera, followed by the lesion group, abundance of genders Ureaplasma, Prevotella e Shuttleworthia. The differences at the species level are extremely necessary for the understanding of the physiological role of the vaginal microbial in the maintenance of autochthonous balance and of pathogenic mechanisms in the development of diseases related to vaginal microbiota. In addition, an understanding of the functionality of the types of CST is necessary to complement what we already know about its structure. Studies are needed to investigate the changes and stability of this microbiota. / O papel da microbiota humana nos processos de saúde e doença tem recebido maior atenção devido à facilidade para a caracterização das comunidades microbianas utilizando os métodos independentes de cultivo, que se baseiam na análise das regiões hipervariáveis do gene 16S rRNA. A microbiota vaginal é habitada por comunidades microbianas que exercem papel importantíssimo para a manutenção da homeostase vaginal e prevenção da colonização por microrganismos patogênicos, mas os mecanismos pelos quais exercem essa influência ainda não são tão bem definidos. Neste contexto, uma compreensão quanto a sua abundância relativa e variações é necessária para o reconhecimento de microrganismos patogênicos potenciais e de processos fisiológicos de proteção dessa microbiota. Este estudo avaliou a diversidade microbiana vaginal em quatro condições distintas: I. microbiota autóctone; II. Microbiota apresentando candidose vaginal; III. microbiota apresentando vaginose bacteriana e; IV. microbiota apresentando lesões pré-malignas e malignas do colo do útero. Amostras cervicais de 187 mulheres foram coletadas, caracterizadas e diagnosticadas a nível molecular quanto à presença de HPV, C. trachomatis, T. vaginalis e N. gonorrhoeae. Amostras HPV+ foram genotipadas em ABI 3500 e a co-infecção do HPV foi relacionada à presença dos outros patógenos e fatores sócio-econômicos e clínicos obtidos através do questionário preenchido pelas voluntárias. Análises de significância e associação foram realizadas através dos Testes Qui-Quadrado de Pearson, Exato de Fischer e Kruskal-Wallis utilizando o R 2.9.0. Após a composição dos grupos, foram montadas quatro bibliotecas de amplicons das regiões V1-V2 do gene 16S rRNA e, posteriormente sequenciadas em plataforma Ion PGM. As sequências geradas foram analisadas utilizando o QIIME e classificadas por comparação no banco Greengenes. A prevalência encontrada para HPV foi de 51,33% sendo que os tipos mais prevalentes foram HPV 16, 58 e 33. As prevalências encontradas para CT e TV foram de 6,42% e 12,83%, respectivamente, sendo observadas co-infecção HPV/CT em 5,20% e HPV/TV em 6,25% das mulheres. O DNA de N. gonorrhoeae não foi encontrado nas amostras. Quanto à estimativa da diversidade microbiana, o número de amostras sequenciado foi suficiente para garantir a cobertura da diversidade total encontrada. A abundância geral revelou predominância do gênero Lactobacillus nos quatro grupos de estudo, seguido pelos gêneros Ureaplasma, Prevotella e Shuttleworthia entre os grupos estudados. A maior diversidade microbiana foi encontrada no grupo Vaginose, tendo como gêneros mais abundantes Prevotella, Shuttleworthia e Megasphaera, seguido pelo grupo Lesão, com abundância dos gêneros Ureaplasma, Prevotella e Shuttleworthia. As diferenças a nível de espécie são extremamente necessárias para o entendimento do papel fisiológico da microbiota vaginal na manutenção do equilíbrio autóctone e de mecanismos patogênicos no desenvolvimento de doenças relacionadas à microbiota vaginal. Além disso, uma compreensão da funcionalidade dos tipos de CST é necessária para complementar o que já sabemos sobre sua estrutura. Estudos são necessários para investigar as mudanças e a estabilidade desta microbiota.

The supraspinal control of descending inhibition of genital reflexes (such as ejaculation) is poorly understood but is important in our global comprehension of how neural signals are integrated to produce sexual behavior, and in our understanding of sexual dysfunction. Sexual dysfunctions, such as premature ejaculation/delayed ejaculation in men, and involuntary vaginal spasms, dyspareunia, and anorgasmia in women, are common. An underlying dysregulation of genital reflexes may produce these dysfunctions, especially in those individuals being treated for depression and anxiety with serotonergic drugs. The nucleus paragigantocellularis (nPGi) of the rat medulla has been described as a descending inhibitory system for genital reflexes in rats, and a homologue is known in humans. Through retrograde tracing of nPGi afferents with the tracer Fluorogold in rats, we found that a number of brain regions implicated in sexual behavior, such as the medial preoptic area, paraventricular nucleus of the hypothalamus, and periaqueductal gray (PAG) provide sexually dimorphic projections to the nPGi, and that many of these regions contain receptors for gonadal steroids and are active during sexual behavior. We also found that excitotoxic lesions of the nPGi with N-methyl-D-aspartate facilitate male sexual behavior by reducing the number of intromissions required for ejaculation, and decreasing ejaculation latency. In females, such lesions attenuated sexual behavior by reducing the amount of time the female spent mating and reducing the reinforcement value of vaginocervical stimulation. Lastly, we found that by removing the source of serotonin to the nPGi (from the ventrolateral PAG) with the serotonergic neurotoxin 5,7-DHT in male rats, we were able to mimic the effects of nPGi lesions and facilitated male sexual behavior indicating that serotonin neurotransmission at the level of the nPGi is critical for genital reflex control. Taken together our results indicate that the nPGi is an important site of integration of internal signals for the regulation of sexual behavior that is sexually dimorphic and under serotonergic control. Our understanding of normal and dysfunction genital reflex control, and possible treatment options in people, is complemented by these results.

No description available.

Source: Masters Abstracts International, Volume: 47-06, page: 3500. Adviser: Kim Waller. Includes bibliographical references.

Thesis (Ph. D.)--University of Washington, 2001. / Vita. Includes bibliographical references (leaves 121-150).

published_or_final_version / Medical Sciences / Master / Master of Medical Sciences

published_or_final_version / Pathology / Master / Master of Medical Sciences

The ecological niche of the vagina contains a large number of different microbes that are constantly interacting with each other and the host. Culture methods have not been sufficient in order to resolve the complexity of the normal vaginal flora. Further, the methods for delineating normal flora from not normal flora are not easily handled and are traditionally not based on culture but on microscopy of elements of the vaginal fluid. In the work presented in this thesis, an international collaboration was established that pin-pointed some of the difficulties in classifying vaginal floras, including staining, sampling, and discordance when lactobacilli are few in number, and that emphasized the importance of the size of the vision field in microscopes. As lactobacilli are prominent members of the normal vaginal flora they need to be carefully classified if further work towards more robust scoring tools is to be achieved. Phenotypic methods have not been able to separate the closely related Lactobacillus species of the vagina. Progress in molecular biology has provided possibilities to characterize these lactobacilli, which are mainly from the Lactobacillus acidophilus group. In this work a large number of strains collected by true random sampling were subjected to RAPD-PCR, TTGE and multiplex PCR for species identification. The major species found were L. crispatus, L. gasseri and L. jensenii and the recently described L. iners. The presence of L. iners has not been detected in previous studies due to its special nutrient requirements. Development of pyrosequencing technology also made it possible to match signatures of the two variable regions V1 and V3 of the 16S rRNA gene of the vaginal lactobacilli and identify them to the species level in a high throughput manner. The study confirmed that the dominating flora in women with normal vaginal flora comprises the four species mentioned previously. Repetitive sampling during IVF-treatment with highly varying oestrogen levels demonstrates changes that possibly occur during changes in the natural life cycle. Furthermore, L. iners was found to be the first species to be established after spontaneously resolved or treated Bacterial Vaginosis. These findings can be of help in developing new strategies for regaining and retaining the normal vaginal flora.

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2007. / Härtill 4 uppsatser.

Diss. (sammanfattning) Umeå : Univ., 2007. / Härtill 4 uppsatser.