mormaeche@dge.gob.pe / Objective: To assess the seroprevalence of hepatitis B virus (HBV), syphilis, and HIV and associated risk factors in pregnant women and their male partners from six indigenous populations of the Peruvian Amazon Basin. Methods: A cross-sectional study was performed in six indigenous populations from the Peruvian Amazon Basin. Blood samples were obtained and tested for HBV (antibodies to the hepatitis B core antigen (anti-HBc) and hepatitis B surface antigen (HBsAg)), for syphilis (rapid plasma reagin and microhemagglutination assay for Treponema pallidum antibodies), and for HIV (ELISA and indirect immunofluorescence test). A survey was also performed to identify associated risk factors. Results: One thousand two hundred and fifty-one pregnant women and 778 male partners were enrolled in the study. The seroprevalence of anti-HBc in pregnant women was 42.06% (95% confidence interval (CI) 39.28–44.85%) and in their male partners was 54.09% (95% CI 50.32–57.86%). The seroprevalence of HBsAg in pregnant women was 2.11% (95% CI 0.78–3.44%) and in their male partners was 3.98% (95% CI 1.87–6.08%). The seroprevalence of syphilis in pregnant women was 1.60% (95% CI 0.86–2.33%) and in their male partners was 2.44% (95% CI 1.22–3.66%). HIV seroprevalence in pregnant women was 0.16% (95% CI 0.02–0.58%) and in their male partners was 0.29% (95% CI 0.04–1.03%). Sexual risk factors were strongly related to blood markers of syphilis and HBV. Conclusions: Hepatitis B was found to be hyperendemic and strongly related to sexual factors, suggesting an important sexual component in the transmission of the disease in the populations studied. Syphilis was found to have an endemicity in pregnant women above the national level and this may be indicative of high mother-to-child transmission. HIV has started to show its presence in indigenous populations of the Amazon Basin and the results suggest the epidemic is concentrated. / Revisión por pares

Periodontal disease is a progressive inflammatory condition that is characterized by the reabsorption of alveolar bone, the destruction of connective tissue, and edentulism. It is caused by a dysbiosis in the oral microbiome as a result of a shift from a Gram-positive aerobic bacterial population, to one that becomes more Gram-negative and anaerobic. Treponema denticola is thought to drive this diseased state based on its role as a keystone periopathogen. A major component of T.denticola’s invasiveness is its motility, which allows the spirochete to penetrate and disseminate through tissues. This motility, which has been seen to be crucial to the invasiveness in other spirochetes through deletion studies, is often regulated by the second messenger, c-di-GMP. In this study, biochemical and biophysical assays were utilized to determine that the predicted diguanylate cyclase TDE0125 converts GTP to c-di-GMP. This elucidates further function of the c-di-GMP regulatory network in T. denticola.

Thesis (Ph. D.)--University of Washington, 2002. / Vita. Includes bibliographical references (leaves 89-111).

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv., 2007. / Härtill 5 uppsatser.

Thesis (Ph. D.)--University of Washington, 1998. / Vita. Includes bibliographical references (leaves [63]-80).

Masters of Science / Background: Infection with HIV results in gradual loss of immunologic functions, especially those mediated by CD4+T helper cells with consequent impairment of the immune response leading to severe manifestations of periodontal disease. The lower the CD4+T lymphocyte cell count or the higher the level of immunosuppression, the higher the incidence of periodontal disease in those patients will be. Putative periodontopathic bacteria namely Porphyromonas gingivalis, Treponema denticola and Tannerella forsythia, commonly referred to as "red complex", and many other bacterial species have been implicated in the initiation and progression of periodontal disease. Objective: The present study tests the association between different CD4+T lymphocyte levels and "red complex" pathogens using BANA, in HIV-positive patients with chronic inflammatory periodontal disease (CIPD). Methods: 120 HIV-positive patients from the infectious disease clinic at Tygerberg hospital participated in the study with a mean age of 33.3 years. The CD4+T lymphocyte counts were obtained from patient's medical records. The six Ramjford teeth were used for evaluating periodontal clinical parameters such as plaque index, gingival index, periodontal probing depth and clinical attachment loss. Subgingival plaque samples were collected and analyzed by the enzymatic BANA test for the detection of the "red complex". Results: The CD4+T lymphocyte mean level was 293.43cells/mm3. Statistically significant associations were found between CD4+T cell counts and probing depth (p= 0.0434) and clinical attachment loss (p= 0.0268). Significant associations were found between BANA with all the clinical indices (p= <0.05). However no association was found between CD4+T cell counts and BANA. Conclusion: HIV-positive patients show a high prevalence of "red complex" pathogens subgingivally. Immunosuppression seems to favour the colonization of these species, resulting in periodontal disease manifestations.

Submitted by Leonardo Freitas (leonardo.hfreitas@ufpe.br) on 2015-04-15T16:03:12Z No. of bitstreams: 2 Dissertaçao SUELY FERREIRA GOMES.pdf: 1499649 bytes, checksum: 4355682f16ea80e03822f54890573e37 (MD5) license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) / Made available in DSpace on 2015-04-15T16:03:12Z (GMT). No. of bitstreams: 2 Dissertaçao SUELY FERREIRA GOMES.pdf: 1499649 bytes, checksum: 4355682f16ea80e03822f54890573e37 (MD5) license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Previous issue date: 2013-04-30 / A Sífilis na gestação, quando não tratada ou inadequadamente tratada, pode ocasionar a Sífilis Congênita, que é um agravo de saúde passível de eliminação como problema de saúde pública. No Brasil, os números de morbidade e mortalidade são preocupantes. Várias políticas de controle de prevenção da doença foram implantadas nos últimos anos exigindo dos profissionais de saúde a adoção de condutas adequadas diante de uma gestante com sífilis, ou seja, a realização do pré-natal de qualidade. Este estudo tem como objetivo analisar os conhecimentos, atitudes e práticas dos médicos e enfermeiros da Estratégia de Saúde da Família na assistência pré-natal das gestantes com Sífilis da cidade do Recife. Trata-se de um estudo descritivo, de corte transversal, com abordagem quantitativa, desenvolvido nas Unidades de Saúde da Família do município. Os principais resultados mostram que os profissionais de saúde, médicos e enfermeiros, possuem um conhecimento parcial com relação a várias ações relacionadas ao diagnóstico, tratamento e acompanhamento da gestante com sífilis, apenas 57% das questões foram respondidas corretamente pelos profissionais. Em relação à adoção de práticas preconizadas pelo Ministério da Saúde, o estudo evidenciou que 59% dos enfermeiros não realizam a prescrição da medicação para tratamento da sífilis na atenção básica. Para um bom desempenho, os profissionais que realizam o pré-natal na atenção básica precisam de capacitação técnica, oferecida nos espaços de educação permanente, mudanças das práticas clínicas, para realização de uma assistência pré-natal integral e mais humanizada, especialmente para às gestantes com sífilis.

Recent bioinformatic analyses identified an operon encoding a potential Type I Secretion System (T1SS) in Treponema pallidum that we hypothesize functions to export key treponemal virulence factors that may contribute to the unique invasiveness and pathogenesis of this spirochete. The membrane fusion protein component (MFP) of T1SSs in other organisms has been shown to play a role in substrate recognition. Hence, the objective of this project is to use the putative MFP, Tp0965, of the potential T. pallidum T1SS to investigate protein-protein interactions with the T. pallidum virulence factor pallilysin (Tp0751) and assess the possibility of the latter being a T1SS substrate. Moreover, protein-protein interactions between Tp0965 and a Treponema phagedenis lysate are investigated with the goal of identifying putative T1SS substrates in this spirochete that could result in the discovery of novel T. pallidum virulence factors via amino acid sequence similarity. Plate-based binding studies and pull-down assays showed a low level of interaction between recombinant Tp0965 and the previously characterized host-component-binding protease, pallilysin, suggesting that the export of this virulence factor could occur via the putative T1SS. Additionally, bioinformatic analyses of the related but cultivable model spirochete T. phagedenis predicted the presence of a potential T1SS homologous to the putative T1SS in T. pallidum. Thus, a more global and unbiased pull-down assay using “bait” Tp0965 and a “prey” T. phagedenis lysate was carried out, followed by mass spectrometric analysis to identify putative novel T1SS substrates with potential homologs in T. pallidum. We successfully identified a T. phagedenis protein, TphBIg, that showed evidence of an interaction with Tp0965. TphBIg seems to possess characteristics of a T1SS substrate suggesting it may be secreted via this system in T. phagedenis. Upon bioinformatic analysis, it was found that TphBIg showed weak amino acid sequence similarity as well as some structural similarity to the T. pallidum protein, Tp0854. Tp0854 is predicted to contain a sialidase and a phosphatase domain with an RTX motif, which is characteristic of some T1SS substrates. Thus, it was hypothesized that if Tp0854 had characteristics of a T1SS, it may interact with Tp0965. Therefore, the phosphatase domain containing the RTX motif was produced recombinantly and plate-based binding studies indeed suggested an interaction with Tp0965, confirming the in silico-predicted interaction. Future experiments to characterize the potential T1SS and substrates in T. pallidum could comprise the functional and structural characterization of the novel putative T1SS substrate, Tp0854. This would include assays to investigate the putative sialidase and phosphatase activities of Tp0854, as well as the identification of Tp0854-Tp0965 interacting sites. Moreover, as a more definite test for T1SS substrate secretion, T. pallidum pallilysin and/or Tp0854 could be expressed heterologously in an E. coli strain harbouring an endogenous T1SS and test for secretion. Similarly, the reconstitution of the T. pallidum putative T1SS in liposomes could be used to further investigate the secretion of pallilysin and/or Tp0854 via this system. Additionally, the optimized unbiased pull-down technique could be further applied to detect more protein-protein interactions within T. pallidum and potentially lead to the identification of more virulence factors that may be secreted via the T1SS. These studies constitute the first investigation of a putative T1SS and substrates within T. pallidum. Thus, insight gained will lead to a better understanding of the mechanisms facilitating T. pallidum host invasion and may reveal new potential vaccine targets to prevent bacterial dissemination and chronic infection. / Graduate

Made available in DSpace on 2014-06-12T23:02:15Z (GMT). No. of bitstreams: 2 arquivo969_1.pdf: 1672515 bytes, checksum: 57a925ef24be8c7dd06dd31514a160d4 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2009 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / O presente trabalho teve por objetivo avaliar a presença dos periodontopatógenos que formam o complexo vermelho (Tannerella forsythia (Tf), Porphyromonas gingivalis (Pg) e Treponema denticola (Td)) e o Aggregatibacter actinomycetemcomitans (Aa) em pacientes portadores de periodontite crônica. A amostra foi constituída de 29 pacientes com diagnóstico clínico e radiográfico de periodontite crônica de acordo com os critérios da AAP (2000). Todos os dentes foram sondados em seis sítios para registro de profundidade, perda de inserção clínica e sangramento após sondagem. As amostras para análise microbiológica foram coletadas dos 4 sítios com maior profundidade de sondagem para cada paciente, totalizando 116 amostras. Estas amostras foram processadas através da técnica de PCR convencional e foram observados os seguintes resultados: 46,6% apresentaram resultado positivo para a bactéria Pg; 41,4% para Tf; 33,6% para Td e 27,6% para Aa. Não se verificou associação significante entre a presença dos periodontopatógenos e as variáveis faixa etária, sexo e sangramento à sondagem. Para a bactéria Pg verificou-se associação significante (p<0,05) com a variável placa visível, e a presença das bactérias Pg e Tf foi mais prevalente (p < 0,05) em bolsas periodontais &#8805; 8 mm. Nos sítios com profundidade &#61619; 8 mm foram observadas com maior freqüência as combinações Pg + Tf (23,2%) e Pg + Tf + Td (20,0%). Foram estatisticamente significantes (p < 0,05) as associações entre a presença simultânea das bactérias Aa + Pg, Aa+ Tf, Pg + Tf e entre Tf + Td. Concluiu-se que as bactérias analisadas, principalmente as do complexo vermelho, estiveram fortemente relacionadas com a periodontite crônica, e que as bactérias Pg e Tf foram mais frequentes em bolsas periodontais profundas

Syfilis är en sexuellt överförbar infektion som orsakas av spiroketen Treponema pallidum subspecies pallidum. Laboratoriediagnostik utgörs i första hand av serologiska test tillsammans med kliniska fynd men på grund av olika faktorer är dessa inte helt pålitliga. Ett flertal olika PCR-metoder utvecklats som påvisar patogenen med hjälp av T-pallidum-specifika gener. En väl studerad gen med hög specificitet är polA-genen som anses vara en mycket robust och känslig metod och lämpar sig väl för att påvisa T. pallidum i kliniska material. Syftet med projektet är att utveckla och optimera en kvalitativ realtids-PCR i singelplex format för verifiering av T. pallidum, för diagnostisering av syfilis. I detta projekt har en realtids-PCR riktad mot polA för detektion av T. pallidum utvecklats. Metodens prestanda utvärderades med en kommersiellt framställd DNA-kontroll med avseende på sensitivitet, specificitet detektions-gräns. När samtliga moment verifierats och fastställts testades metoden på kliniskt material från sår. Den T. pallidum-specifika PCR-analysen visade att det inte förekom korsreaktivitet mot andra agens som förväntas finnas i sår från patient med misstänkt syfilis. Metoden uppvisade en känslighet vid spädning 1:100 (cirka 13 kopior/µl) och en precision på 36,3±0,8 Ct. Det kliniska provet visade på förekomst av T. pallidum dock är känsligheten något sämre än existerande referensmetod. Metoden kan användas i rutindiagnostik av T. pallidum dock bör utfallet från extraktion av DNA från sår studeras ytterligare för att eventuellt öka utbytet och detektera lägre koncentrationer i kliniskt material. / Syphilis is a sexually transmitted infection caused by the spirochete Treponema pallidum subsp. pallidum. Laboratory diagnostics consist primarily of serological tests together with clinical findings for different reasons these methods are not reliable. A variety of PCR methods has been developed that target the pathogen using T. pallidum-specific genes. A well-studied gene with high specificity is the polA gene which is considered very robust and sensitive method and well suited for detection of T. pallidum in clinical materials. The aim of the project is to develop and optimize a qualitative real-time PCR in single-plex format for the verification of T. pallidum, for the diagnosis of syphilis. A real-time PCR targeting polA was developed for detection of T. pallidum. An evaluation of the method's performance was done with a commercially produced DNA control with regards to sensitivity, specificity detection limit and precision. When all test where verified and established, the method was tested on clinical material from ulcers. The results of the T. pallidum-specific PCR-assay showed that there was no cross-reactivity to other agents that are expected to be in ulcers from patients with suspected syphilis. The method showed a sensitivity at dilution 1:100 (about 13 copies/μl) and a precision of 36.3 ± 0.8 Ct.The clinical specimen showed presence of T. pallidum, however, the sensitivity was not as good than the existing reference method. The method can be used in routine diagnostics of T. pallidum, however, the outcome of extraction of DNA from ulcers should be further studied in order to increase the yield and detect lower concentrations in clinical material.