Thesis (Ph.D.) -- University of Adelaide, Dept. of Paediatrics, 2004. / "March 2004" Includes bibliographical references (leaves 213-272).

Diss. (sammanfattning) Uppsala : Uppsala universitet, 2009. / Härtill 5 uppsatser.

Thesis (M.S.)--University of Missouri-Columbia, 2004. / "December, 2004." The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Includes bibliographical references.

Thesis (Ph. D.)--University of Washington, 2006. / Vita. Includes bibliographical references (leaves 78-85).

Made available in DSpace on 2015-07-08T12:28:19Z (GMT). No. of bitstreams: 2 6.pdf: 926522 bytes, checksum: f6e46b30951c9cffb7632bb9dd259770 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2013 / Fundação Oswaldo Cruz. Instituto Nacional de Controle de Qualidade em Saúde / As bactérias do gênero Haemophilus, família Pasteutelleaceae têm ainda na espécie influenzae influenzae (Hi)a mais importante nas infecções humanas. Hi, que inclui os sorotipos capsulares (a-f) e os NT é responsável por diversos quadros infecciosos. Haemophilus influenzae b (Hib) era um dos principais responsáveis pelos casos de meningites em diversos países do mundo. Estava também associado a outras infecções graves como epiglotite, artrite séptica, bacteremia, pneumonia e septicemia, principalmente em crianças. As doenças associadas ao Hib são preveníveis pela vacina conjugada formada do PRP e uma proteína carreadora, incluída no PNI/MS em agosto de 1999. Após a introdução da vacina conjugada contra o Hib houve redução expressiva das doenças causadas pelo Hib, nos diversos países que introduziram a vacina em seus calendários de imunização, porém, os outros sorotipos, inclusive os HiNT passaram a ser isolados com maior frequência como agente infeccioso de meningites sendo também atualmente um dos principais agentes etiológicos da Otite Média Aguda (OMA), o que o torna alvo de importantes pesquisas para novas vacinas. A colonização desta é fundamental para que ocorra a infecção e, crianças de creches e orfanatos apresentam variáveis taxas desta bactéria. A importância deste estudo continuando uma linha de pesquisa sobre o Hi no INCQS-FIOCRUZ, com ênfase no estudo de cepas isoladas após a implantação da vacina está associada à necessidade de verificar a situação de portadores, em uma população fechada (creche) para contribuir com o conhecimento da situação do Hi após treze anos da introdução da vacina conjugada contra o Hib no Brasil, que reduziu, mas não impediu a ocorrência da doença. O objetivo principal deste estudo foi determinar a ocorrência de Hi em crianças frequentadoras de uma creche em Jacobina, Bahia. Realizou-se coleta de material da nasofaringe de 73 crianças. / The Bacterial genus Haemophilus is inclued in the family Pasteutelleaceae and the influenzae specie (Hi) is the most important in human infections. Hi includes capsular serotypes (a-f) and non-capsulated strains (NT), which are responsible for many infections. Haemophilus influenzae b (Hib) has been one of the main cause of meningitis in many countries worldwide. It has been also associated with other severe infections such as epiglottitis, septic arthritis, bacteremia, pneumonia and septicemia, mainly in children. Since 1988 these infections are preventable by Hib conjugate vaccine consisting of PRP and a carrier protein, and it has been included in the national vaccination program (PNI/ MS) in 1999. After the introduction of Hib conjugate vaccine, diseases caused by Hib had significantly decreased in several countries where the vaccine was introduced into their immunization schedules however, other serotypes, including HiNT, has been isolated with greater frequency as agents of infectious meningitis which is currently one of the main etiological agents of Acute Otitis Media (AOM) and the subject of important researches on new vaccines. Colonization is essential to start the infection, and children from daycare centers present variable rates of the bacteria in their nasopharynx. The importance of this study on an ongoing line of research at INCQS-FIOCRUZ with emphasis on the study of strains isolated after the introduction of the vaccine, is associated with the monitor of carriers in a closed population (daycare) to contribute to the knowledge of the circulation of Hi after thirteen years using conjugate vaccine against Hib in Brazil, which reduced but did not prevent the occurrence of the disease. The main objective of this study was to determine the occurrence of Hi in children attending a daycare center in Jacobina, Bahia. For the study we collected material from the nasopharynx of 73 children. We found a colonization rate of 78.08% and all isolates were classified as HiNT. Biotype III was the most prevalent. The study of antimicrobial susceptibility showed ampicillin-resistant strains producing and non-producing β-lactamase. We also observed high rates of resistance to ampicillin (41.8%) and trimethoprim+sulfametoxazol (67.2%). The gene blaTEM, which is primarily responsible for ampicillin resistance was found in 81.8% of the strains. The blaROB gene, was not observed among the strains analyzed. The PBP3 gene of βLNAR samples was sequenced and changes were observed, the major changes were in positions 547V → 547I found in three samples, 422N→ 422S and 273S→ 273F. Strain susceptibility was evaluated against amoxicillin/clavulanic acid, ceftriaxone, rifampicin, chloranphenicol and were susceptible. This study confirms the efficacy of the vaccine against Hib and points to the relevance to vigilance HiNT, as presented high percentage of colonization and resistance to some antimicrobials.

Orientador: Marcelo Lancellotti / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-23T12:32:22Z (GMT). No. of bitstreams: 1 Cury_GiseleCristianeGentile_D.pdf: 5956781 bytes, checksum: b28217e4d95beb4734f0357ed4181696 (MD5) Previous issue date: 2013 / Resumo: O resumo poderá ser visualizado no texto completo da tese digital quando for liberada / Abstract: The abstract is available with the full electronic document when available / Doutorado / Bioquimica / Doutora em Biologia Funcional e Molecular

No description available.

Orientador: Marcelo Lancellotti / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-22T07:21:21Z (GMT). No. of bitstreams: 1 Varela_JuliaNogueira_M.pdf: 1464845 bytes, checksum: 930cccae996588333b99f1aaa50988c0 (MD5) Previous issue date: 2013 / Resumo: Haemophilus influenzae é uma bactéria causadora de doenças tipicamente associadas ao trato respiratório superior e inferior. Tal bactéria é classificada em linhagens capsuladas e não capsuladas - as não tipadas. As grandes responsáveis por patogenias mais severas são as capsuladas, especialmente as do sorotipo b, a existência de uma vacina para somente esse sorotipo, faz com que ocorra uma emergência de casos com H. influenzae não tipado - NTHi. A crescente resistência a antibióticos dessa bactéria está associada à plasmídios de resistência, bem como sua competência natural. A presença desses patógeno é maior em países nos quais não existe acesso a vacina, devido ao alto custo da mesma, que acabam utilizando antibióticos mais acessíveis como o cloranfenicol no tratamento. Esse trabalho estudou a transferência horizontal do gene ompP2 em diversas cepas de H. influenzae com a ajuda de nanopartículas de óxido de grafeno. Essas nanopartículas mimetizam uma atmosfera rica em partículas suspensas como as grandes cidades e zonas de agricultura precoce, já que, nesses locais ocorrem com maior frequência mutações e adaptações desse patógeno. Quando as nanopartículas encontravam-se no meio de cultura, verificou-se um aumento da taxa de transformação dessas bactérias. Assim como uma modificação no padrão de adesão celular das bactérias mutadas quando comparadas com as selvagens em linhagens celulares distintas e expostas ao antibiótico de resistência, levando a um aumento da taxa de adesão das cepas mutadas com relação às cepas selvagens. Como esse gene é e de possível aquisição entre cepas de H. influenzae em seu ambiente natural seria possível utilizá-lo para obtenção de uma proteína recombinante, com possível antigenicidade. Uma vez que a taxa de adesão aumenta com a presença do mesmo, levando a uma possível nova vacina que também protegeria contra cepas não tipadas e não somente capsuladas / Abstract: Haemophilus influenzae is a bacteria that causes diseases typically associated with the upper and lower respiratory tract. Their strains are divided in capsulated and non-capsulated - the non typable. The major responsible for more severe cases are the capsulated types, specially the b type. The existence of a vaccine for the serotype b, allows the emergence of cases of non typable H. influenzae - NTHi. The growing resistance is associated with resistance plasmids, and with its natural competence, that enables the bacteria to acquire DNA fragments between it's' species. Since this pathogen is common in countries that there is no access to this vaccine, therefore the use of accessible and cheaper antibiotics, such as chloramphenicol for treatment is. This work studied the horizontal transference of the ompP2 gene from multiresistant strains of H. influenzae, with the aid of grafen oxide nanoparticles, that mimesis an atmosphere rich in suspended particles, such as great urban areas and ancient agricultural zones. In these environments a great frequency in mutation and adaptations of these bacteria is verified. When we look at the adhesion patterns of these bacteria we can see that it is modified when they are mutated and exposed to the resistance antibiotic. Leading to an augmentation of the adhesion patterns when we compare to the wild strains. Since this gene was present in all strains and it was of easy acquisition between strains, it would be possible to use it to obtain a recombinant protein with likely antigen properties. Because the adhesion tax enhances with the presence of this gene. Leading to a possible new vaccine target, for NTHi and capsulated strains also / Mestrado / Fármacos, Medicamentos e Insumos para Saúde / Mestra em Biociências e Tecnologia de Produtos Bioativos

OBJECTIVE: The principal objective of this study was to measure various aspects of immunity in children with invasive infections due to Haemophilus influenzae type b and Staphylococcus aureus. These serious infections are a significant cause of childhood morbidity and mortality in all populations and affect healthy as well as compromised children. Evidence suggests that imbalances or deficiencies in certain aspects of immunity such as IgG subclasses, the capacity to make specific subclass antibodies, antibody affinities, complement isotypes, immunoglobulin allotypes or mannose binding protein may place certain children at risk for developing invasive disease. Investigation of these factors in a group of children with infection necessitated that normal ranges be established for children of comparable ages from the same population. A secondary objective of this study has therefore been to establish normal percentiles for the IgG subclasses in age, race and sex matched healthy controls. METHODS: Patients admitted to the Red Cross War Memorial Children's Hospital with septic meningitis due to Haemophilus influenzae type b and osteomyelitis/septic arthritis due to Haemophilus influenzae type b or Staphylococcus aureus formed the study population. Section A of this thesis describes the methods for establishing, validating and standardizing ELISAs for measuring the IgG subclasses (lgGl, IgG2, IgG3 and IgG4) and subclass antibodies specific to Haemophilus influenzae polyribosylribitol phosphate, Staphylococcus aureus teichoic acid and tetanus toxoid. The relative affinity of antibodies in these ELISAs was determined by the incorporation of diethylamine (DEA). In order to determine the immunoglobulin allotypes ELISAs were developed to measure the G1m(f), G2m(n) and Km(3) allotypes. The frequency of these allotypic markers in the different ethnic groups was established. The relationship between immunoglobulin allotypes and IgG subclass values were investigated in both patient and control groups. RESULTS: ELISA assays to measure IgG subclasses; IgG, IgG 1 and IgG4 tetanus toxoid antibodies; IgG, IgG 1 and IgG2 H. influenzae type b polyribosylribitol phosphate capsular polysaccharide antibodies; IgG, IgG1 and IgG2 S. aureus teichoic acid antibodies and G1m(f), G2m(n) and Km(3) allotypes were successfully established. Where possible the assays were standardized with reference sera and specimens were exchanged with international laboratories. Age, race and sex related percentile charts and tables of normal ranges for IgG and IgG subclasses of Black and Coloured children were established. The IgG and IgG 1 values were higher than those previously reported for children in developed countries. Black children with H. influenzae meningitis had significantly lower IgG 1, IgG2 and IgG3 levels compared to the controls and although similar trends were seen for IgG and IgG4 levels they were not statistically significant. Coloured children with H. influenzae meningitis and Coloured and Black children with H. influenzae osteomyelitis/septic arthritis also showed a similar tendency of lower IgG and IgG subclass levels than the controls but these trends were also not significantly different. All patients responded to tetanus toxoid antigen suggesting normal immunocompetence to protein antigens. H. influenzae type b capsular polysaccharide antibodies were low in children with H. influenzae type b meningitis and osteomyelitis/septic arthritis and did not increase during the illness. IgG and IgG 1 teichoic acid antibodies were raised in patients with S. aureus osteomyelitis/septic arthritis although no further rise in these antibodies was seen when measured several weeks after the illness. The antibody affinity ELISAs showed that IgG 1 tetanus toxoid antibody had a greater affinity than IgG4 tetanus toxoid antibody, the IgG 1 and IgG2 H. influenzae capsular polysaccharide antibodies were of similar affinity and the IgG 1 teichoic acid antibody was of higher affinity than the IgG2 antibody. The G1m(f) and G2m(n) positive allotypes were uncommon in Black but common in the Coloured populations whereas Km(3) was common in both groups. There was a significantly decreased frequency of the G2m(n) positive allotype in Coloured patients with H. influenzae type b meningitis and H. influenzae type b osteomyelitis/septic arthritis which was not found in patients with S. aureus osteomyelitis/septic arthritis. In both Coloured and Black children with H. influenzae meningitis there was a significantly decreased frequency of the Km(3) allotype. No differences in C4 isotypes and mannose binding protein levels were evident in the patient and control groups. CONCLUSION: This study has developed simple, specific and reproducible ELISAs to measure IgG subclasses and subclass antibodies specific to tetanus toxoid, H. influenzae polyribosylribitol phosphate and S. aureus teichoic acid. Age, sex and race related normal ranges for IgG subclasses in the local Black and Coloured populations have been established. Black children with H. influenzae type b meningitis had significantly lower IgG 1, IgG2 and IgG3 levels compared to the controls. There was a clear association between a decrease of the G2m(n) allotype and the Km(3) allotype and susceptibility to invasive infections caused by H. influenzae.

Nosocomial outbreaks of infection due to non-typeable Haemophilus influenzae (NTHi) are rarely described. There are a few published reports that suggest that elderly patients with underlying pulmonary disease are at risk and that person-to-person spread is key to disease transmission. During the summer months of 2005, we documented an outbreak of NTHi infections in a Veterans Affairs nursing home. Thirteen patients developed conjunctivitis or lower respiratory infection involving a β-lactamase-negative biotype III NTHi isolate, with an indistinguishable SmaI macrorestriction pattern. Patients were elderly males usually with underlying cardiac and pulmonary disease. A case-control study failed to demonstrate any specific significant risk factor for NTHi infection and there was no evidence of spatial clustering of cases within the nursing home. A random throat culture survey involving nursing home patients during the outbreak showed that only one of 19 persons was colonized with NTHi. The outbreak concluded following appropriate treatment and an emphasis on universal and respiratory droplet precautions. All patients recovered and a specific inciting event for the outbreak was never defined. Literature review revealed a spectrum of responses to nosocomial NTHi infections and a lack of consensus regarding the infection control approach towards NTHi outbreaks.