Os surtos de doença de Chagas aguda estão relacionados ao consumo de comidas e bebidas contaminadas por fezes de triatomíneos contendo tripomastigotas de T. cruzi, tornando assim a infecção oral uma importante via de transmissão. Ambas as infecções vetoriais e orais desencadeiam importantes manifestações cardíacas nos hospedeiros e que estão relacionadas a uma desregulada resposta imune. O objetivo deste estudo foi avaliar a parasitemia sanguínea durante a fase aguda da infecção, a produção de macrófagos peritoneais, alterações nas sub-populações de linfócitos CD4+/CD8+, citocinas Th1 e Th2, concentração de nitrito e a histopatologia cardíaca. Um grupo de ratos Wistar foi intraperitonealmente infectados com 1X105 com cepa Y de tripomastigotas sanguíneos de T. cruzi e outro grupo de ratos Wistar foi infectado oralmente com 8X105 de tripomastigotas metacíclicos da mesma cepa. A infecção intraperitoneal desencadeou um aumento estatístico no aumento do número de parasitas, macrófagos peritoneais, proliferação de células T CD4+ e CD8+, aumento na concentração de NO, IL-12 e elevado foco inflamatório cardíaco quando comparado à infecção oral. Entretanto nossos resultados demonstraram que a infecção oral pode levar a uma doença sistêmica e que as diferentes vias de inoculação promovem respostas imunes distintas. / Acute Chagas disease outbreaks are related to the consumption of food or drink contaminated by triatomine feces containing trypomastigotes of T. cruzi, thus making oral infection an important route of transmission. Both vectorial and oral infections trigger important cardiac manifestations in the host that are related to a dysregulated immune response. The aims of this work were to evaluate the number of blood trypomastigotes during the acute phase of infection, peritoneal macrophages, possible alterations of lymphocyte CD4+/CD8+ sub-populations, Th1 and Th2 cytokines, nitrite concentrations and cardiac histopathology. One group of male Wistar rats was intraperitoneally infected with 1X105 blood trypomastigotes of the T. cruzi Y strain, and another group of Wistar rats was orally infected with 8X105 metacyclic trypomastigotes of the same strain. The intraperitoneal infection triggered statistically enhanced parasite numbers, peritoneal macrophage numbers, proliferation of CD4+ and CD8+ T cells, increased concentrations of NO and IL-12 and elevated cardiac inflammatory foci when compared to the oral infection. Therefore, our results demonstrate that oral infection can lead to systemic disease and that the different inoculation routes promoted distinct immune responses.

According to recommendations from the Facility Guidelines Institute (FGI) of the American Institute of Architects (AIA), World Health Organization (WHO) and Center for Disease Control and Prevention (CDC), a common engineering approach to isolation room design is to maintain the air ventilation rate at a minimum of 12 air changes per hour (ACH) for mixing and dilution, and a negative pressure in the room to direct airflow inwards, instead of leaking outwards. / In collaborations with physicians in the Respiratory Division and the Intensive Care Unit (ICU) at the Chinese University of Hong Kong (CUHK), a series of experiments were carried out to verify the ventilation performance of an All room at the Princess Margaret Hospital (PMH). Experiments investigated the effects of ACH, the control of airflow direction, the air tightness of the automatic swing door and the application of positive pressure ventilation procedures, such as high flow rate oxygen masks, jet nebulizers and NPPV. These were extensively tested in two different isolation rooms of the Prince of Wales Hospital (PWH) and PMH, under common clinical circumstances and environmental conditions. / Many patients with severe respiratory infection require supportive therapy for respiratory failure. Common interventions involve supplemental oxygen to improve tissue oxygenation. In the worst scenario, mechanical ventilation via non-invasive positive pressure ventilation (NPPV) may be required. Since a large amount of aerosols is generated during these interventions, there is a great risk of spreading infectious aerosols from the respiratory tract of the patient to the surrounding environment. / The aerodynamic data in this thesis infonns architects and engineers on how to improve the hospital ward ventilation design so as to avoid aerosol and ventilation leakage. Ultimately, it is hoped that this work may play a role in preventing devastating nosocomial outbreaks in the future. / The design of airborne infection isolation (AII) room has become one of the major research domains following the emergence of the global concern of acute respiratory diseases in this century. These include severe acute respiratory syndrome (SARS) in 2003, H5N1 avian influenza, and pandemic influenza H1N1 in 2009. All of which have claimed thousands of lives. Even with the current stringent design and practice guidelines, nosocomial infection of healthcare workers (HCWs) and inpatients continues to occur. This implies that there might be limitations in current isolation ward designs. / The experiments implemented a high-fidelity human patient simulator (HPS) which could be programmed with different lung breathing conditions and oxygen flow rate settings. The patient exhaled air dispersion distances and airflow patterns were captured in detail with a non-intrusive, laser light sheet, smoke particle scattering technique, designed for this thesis. Thin laser light sheets were generated by a high energy YAG laser with custom cylindrical optics. Smoke concentration in the patient exhaled air and leakage jets was estimated from the intensity of light scattered, which was then expressed as nonnalized particle concentration contours using computer programs developed for this study. / The study quantitatively revealed the distinctive patient exhaled airflow patterns and the extent of bioaerosol, generated directly from the patient source with the application of different oxygen delivery interventions for different patient lung conditions and oxygen flow rates. It was found that contamination was more critical during the administration of oxygen therapies, which is common in clinical circumstances. Source control is therefore the most efficient and effective approach to the reduction and even elimination of patient exhaled bioaerosol contaminants. Thus, when working in an isolation room environment, full preventive measure should be taken and it is essential to consider the location of mechanical vents and the patient exhaled airflow patterns. It has also been shown in experiment that applications of bacterial viral filter could be a solution to the problem. / Chow, Ka Ming. / Advisers: Puay Peng Ho; Jin Yeu Tsou. / Source: Dissertation Abstracts International, Volume: 73-09(E), Section: A. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 115-147). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. Ann Arbor, MI : ProQuest Information and Learning Company, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Les Bradyrhizobium photosynthétiques sont capables d'induire la formation de nodules fixateurs d'azote chez certaines légumineuses du genre Aeschynomene. La découverte récente que certaines de ces souches ne possèdent pas les gènes canoniques nodABC indique l'existence d'un nouveau processus symbiotique rhizobium-légumineuse indépendant des facteurs Nod. L'objectif de ce travail de thèse a consisté à avancer dans la compréhension des mécanismes mis en jeu lors de cette nouvelle interaction. Dans un premier temps, à travers différentes approches cytologiques, le processus par lequel la bactérie infecte la plante en l'absence de facteurs Nod a été décrit. Dans un deuxième temps, afin de mettre en évidence les bases moléculaires de cette interaction, une banque de 15 000 mutants Tn5 de la souche ORS278 a été criblée sur plante. Ce criblage a permit l'identification de plus d'une centaine de gènes bactériens intervenant durant le processus symbiotique. Les résultats obtenus nous ont conduits à proposer un modèle dans lequel la mise en place de la symbiose Nod-indépendante impliquerait, d'une part, la synthèse bactérienne d'une cytokinine permettant le déclenchement de l'organogenèse nodulaire, et d'autre part, d'autres signaux bactériens intervenant dans l'étape de reconnaissance avec la plante hôte. Enfin, nous avons mis en place une technique de transformation génétique d'Aeschynomene et validé cet outil à travers l'étude de l'expression hétérologue de la noduline précoce MtENOD11. Il peut à présent être envisagé de conduire des études fonctionnelles sur Aeschynomene en vue de caractériser la voie de signalisation Nod-indépendante. / The photosynthetic Bradyrhizobium are able to induce the formation of nitrogen-fixing nodules in some legumes of the Aeschynomene genus. The recent discovery that some of these strains lack the canonical nodABC genes indicates the existence of a new symbiotic rhizobium-legume process that is independent of Nod factors. The aim of this work was to improve our understanding of the mechanisms involved in this new interaction. First, through various cytological approaches, the process by which the bacterium infects the plant in the absence of Nod factors has been described. Second, in order to decipher the molecular basis of this interaction, a library of 15,000 Tn5 mutants of the ORS278 strain was screened on plant. This screening allowed the identification of about one hundred bacterial genes involved in this symbiotic process. These results led us to propose a model in which the establishment of the Nod-independent symbiosis involves, on one han d, the synthesis of a bacterial cytokinin that triggers nodule organogenesis, and on the other hand, others bacterial signals that permit the recognition with the host plant. Finally, we developed a genetic transformation procedure of Aeschynomene and we validated this tool by studying the heterologous expression of the early nodulin MtENOD11. Now, functional studies on Aeschynomene are possible to permit the characterization of the Nod-independent signaling pathway.

The aim of this study was to determine the prevalence of extended-spectrum b-lactamase (ESBL)- producing Klebsiella pneumoniae isolated from urinary tract and bloodstream infections in a rural hospital in Manhic¸a, Mozambique. ESBLs were investigated among ceftriaxone-non-susceptible K. pneumoniae clinical isolates recovered between 2004 and 2009. Characterisation of blaCTX-M, blaSHV, blaOXA and blaTEM genes was performed by PCR and sequencing. Epidemiological relationships were established by phylogenetic analysis, repetitive extragenic palindromic PCR (REP-PCR), pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST), whilst plasmid transferability was evaluated by conjugation. In addition,the presence of class 1 and 2 integrons was studied.A total of 19 K. pneumoniae were analysed. The blaCTX-M-15 gene was found in all strains. Other ESBL genes were found concomitantly, including blaSHV-5, blaSHV-2, blaSHV-2A, blaSHV-12 and blaSHV-38. In addition, other b-lactamases such as blaTEM-1 and blaOXA-30 were also detected. REP-PCR identified 15 different epidemiological profiles. MLST analysis also showed great variability of sequence types. The blaCTX-M-15 gene showed a high transfer capacity. The presence of class 1 integrons was high. High levels of multidrug resistance were also found. In conclusion, these data show the dominance of the CTX-M-type ESBL, particularly CTX-M-15, supporting its worldwide dissemination, including in areas with limited access to third-generation cephalosporins. This finding is a matter of concern for clinical management as third-generation cephalosporins are an alternative for treating severe cases of multidrug-resistant infections in this community. / Revisión por pares

Introdução: A infecção de sítio cirúrgico representa a terceira causa de infecção relacionada à assistência à saúde, que mais acomete pacientes na Europa e no Brasil. Considerando as altas hospitalares cada vez mais precoces e que grande parte das infecções do sítio cirúrgico se manifestará até o sétimo dia após a cirurgia, a vigilância pós-alta se faz extremamente necessária, uma vez que sua não realização pode levar a subnotificação, dificultando as ações de prevenção e controle. Existe carência, até o momento, de ferramentas que tenham sido validadas para a identificação de potenciais casos de ISC durante a vigilância pós-alta. Objetivo: Criar e validar um instrumento para a detecção pós-alta de potenciais casos de ISC. Método: Estudo psicométrico para a criação e validação de um instrumento, empregando-se as etapas de validação de conteúdo, critério e constructo. Resultado: O instrumento, após validação de conteúdo com juízes especialistas, apresentou coeficiente de validade de conteúdo total igual a 0,87. Para a validação de critério e constructo o instrumento proposto foi aplicado em uma amostra de 100 pacientes e comparado ao exame físico por profissional de saúde, observando-se coeficiente alfa de Cronbach igual a 0,87; Kappa de Cohen igual a 0,83; sensibilidade igual a 76,4%; especificidade de 100%; valores preditivo negativo de 92,5%, e preditivo positivo igual a 100%; precisão de 94%. Conclusão: O instrumento proposto foi validado e demonstrou ser uma ferramenta útil para detecção de possíveis casos de infecção de sítio cirúrgico na vigilância pós-alta. / Introduction: Surgical site infection (SSI) represents the third cause of healthcare-associated infections, which affects patients in Europe and Brazil. Considering hospital discharges that are becoming more precocious and that most SSI will manifest themselves up to the seventh postoperative day, postdischarge surveillance is extremely necessary, since its failure to perform can lead to underreporting, making it difficult to prevention and control actions. There is a lack, until now, of tools that have been validated for the identification of potential cases of SSI during postdischarge surveillance. Objective: To create and validate an instrument for the postdischarge surveillance of potential SSI cases. Method: a psychometric study for the creation and validation of an instrument, using content, criterion and construct validation. Result: The instrument, after validating content with expert judges, presented coefficient of total content validity equal to 0.87. For the validation of criterion and construct the instrument was applied in a sample of 100 patients. Comparing the instrument to the physical examination by the health professional, Cronbachs Alpha was equal to 0.87; Cohen\'s Kappa was equal to 0.83; sensitivity equal to 76.4%; specificity of 100%; negative predictive values of 92.5%, and, positive predictive equal to 100%. The accuracy of the instrument was 94%. Conclusion: The proposed instrument was validated and constitutes a useful tool to detect possible cases of Surgical Site Infection in postdischarge surveillance.

Le diabète sucré est un problème majeur de santé publique. L’une de ses principales complications est l’ulcère du pied (UPD) à l’origine d’infections et de nombreuses amputations. Ces plaies sont majoritairement polymicrobiennes. La principale bactérie isolée est Staphylococcus aureus (SA) qui pose d’importants problèmes en thérapie courante. Bactérie commensale notamment des fosses nasales, elle est un vrai pathogène au niveau des UPD. La compréhension du passage du commensalisme à la pathogénicité reste peu connue. Or récemment une souche colonisante de SA présentant un faible potentiel de virulence, et isolée sur un UPD a été identifiée. Cette souche présente un phage (ROSA-like) inséré dans son génome dont la stabilité a été démontrée. Ce phage est responsable de la faible virulence de cet isolat. Ce travail de doctorat a permis : i) de mettre en évidence l’implication du phage ROSA-like dans la dérégulation des mécanismes d’acquisition du fer, mécanismes essentiels pour la survie du SA ainsi que dans le métabolisme global de cette souche. Ces résultats ont confirmé le rôle clé du phage dans la baisse de la virulence du SA colonisant ; ii) à partir d’un modèle in vitro d’infections chroniques mimant les conditions rencontrées dans les UPD, d’étudier le comportement de souches cliniques de SA (colonisantes/infectantes) en fonction des différentes conditions environnementales : hyperglycémie, anaérobie, antibiotiques (linézolide et vancomycine). Les résultats ont montré qu’une exposition prolongée (plus de 24 semaines) à ces conditions de stress notamment aux antibiotiques, réduisait la virulence des souches de SA ; iii) d’évaluer la virulence des SA en présence d’autres bactéries qui l’entourent, élément essentiel du polymicrobisme de l’UPD. Ce travail s’est particulièrement intéressé aux interactions existant entre SA et Helcococcus kunzii, bactérie fréquemment isolée en association avec le SA dans les UPD. Il a montré pour la première fois une baisse de la pathogénicité des souches de SA par une bactérie commensale non virulente. / Diabetes mellitus is a global problem. One of the main complications is diabetic foot ulcer (DFU) which can evolve towards infection and induces lower limb amputations. Those wounds are mostly polymicrobial and Staphylococcus aureus (SA), the most prevalent pathogen isolated in this situation causes numerous therapeutic problems. This bacterium, known to be a commensal organism present notably in nasal cavity, is a real pathogen in DFU. However the comprehension between the commensalism and the pathogenicity of SA remains still unknown. Recently, a colonizing SA strain with very low virulence potential was discovered in DFU. This strain possesses a phage named Rosa-like responsible of its low virulence.This work helped us: i) to highlight the involvement of the Rosa-like phage in the disregulation of iron uptake mechanisms, the main source for SA survival and its global metabolism. The obtained results confirm the key role of the phage in the decrease of the colonizing SA strain virulence; ii) to study the behaviour of clinical SA strains (colonizing/infecting) using an in vitro chronic infection model miming different environmental conditions encountered in DFU: hyperglycemia, anaerobic condition, antibiotics (linezolid and vancomycin). Results showed that prolonged exposure (up to 24 weeks) to those stress conditions notably antibiotics reduced SA strains virulence; iii) to evaluate SA virulence in presence of other bacteria present in DFU. Particularly, in this work a focus was performed in the interaction between SA and Helcococcus kunzii, gram positive bacteria frequently isolated with SA in DFU. It was shown for the first time a decrease of SA pathogenicity by a commensal non virulent bacterium.

A crescente preocupação sobre as infecções relacionadas à assistência à saúde levou a Organização Mundial da Saúde a promover a criação da Aliança Mundial para a Segurança do Paciente, enfatizando ações básicas de controle de infecções. A redução de sua incidência implica em proteção para o paciente, profissionais e instituições, favorecendo a prestação da assistência livre de riscos. Dentre as mais frequentes, a infecção do sítio cirúrgico deve ser compreendida como elemento qualificador do cuidado e, os dados relacionados, o ponto de partida para as ações preventivas e de controle. Os hospitais têm dificuldades para realizar a vigilância epidemiológica pós-alta, tarefa complexa haja vista a tendência de redução do tempo de internação. Conhecer dados sobre a readmissão contribui para melhorar os dados de vigilância pós-alta. Foi realizado um estudo exploratório com o objetivo de descrever a epidemiologia das readmissões por infecções de sítio cirúrgico em pacientes em um hospital público de Belo Horizonte (MG). Foram avaliados 98 registros médicos e da Comissão de Controle de Infecção Hospitalar referentes aos indivíduos readmitidos por este motivo entre janeiro/2008 e dezembro/2009. Os resultados demonstraram que um quarto dos pacientes que desenvolveu infecção do sítio cirúrgico na instituição necessitou de nova internação. A idade média dos indivíduos foi de 57,2 anos; mais da metade apresentava diabetes mellitus, e um terço, hipertensão arterial sistêmica; no entanto, o risco anestésico para a maioria foi classificado como baixo. As infecções ocorreram com maior frequência em cirurgias limpas e potencialmente contaminadas, e as especialidades com maior número de pacientes readmitidos foram ortopedia, com mais da metade dos procedimentos relacionados à correção de fraturas, e cirurgia geral, na qual as hernioplastias/rafias e colecistectomias predominaram. Metade dos indivíduos recebeu biomaterial em suas intervenções. Os tempos cirúrgicos e de internação não diferiram dos estudos encontrados na literatura. Praticamente todos os pacientes utilizaram antibioticoprofilaxia, segundo protocolo da instituição. Os primeiros sinais e sintomas surgiram, em média, após 33,2 dias, e metade dos infectados teve o diagnóstico nos primeiros 30 dias de pós-operatório. A classificação das infecções mostrou que metade dos pacientes apresentou infecção de órgãos e cavidades e, para 60% deles, foram necessárias novas intervenções cirúrgicas, além do tratamento antimicrobiano. O micro-organismo predominante nas infecções ortopédicas foi Staphylococcus aureus e nas infecções da cirurgia geral Escherichia coli, ambos com perfil de resistência abaixo daqueles encontrados na literatura. O estudo permite concluir que a vigilância pós-alta e o monitoramento das taxas de readmissão contribuem para o redimensionamento do problema e definição de ações pontuais para seu controle. / The growing concern about infections related to health care led the World Health Organization to promote the creation of the World Alliance for Patient Safety, emphasizing basic actions to infection control. The reduction in the incidence of surgical site infection implies protection for patients, professionals and institutions favoring the assistance free of risks. Among the most frequent, surgical site infection should be understood as a qualifying element of care, and related data, the starting point for preventive and control measures. Hospitals have difficulties in achieving post-discharge surveillance, complex task given the trend of reduced hospital stay. Knowledge of the of readmission rates helps to improve reporting of post-discharge surveillance. An exploratory and descriptive study was conducted in order to describe the epidemiology of readmissions for surgical site infections in patients in a governmental hospital in Belo Horizonte (MG). We evaluated 98 medical records and the reports of the Hospital Infection Control Committee related to readmitted patients for this reason among january/2008 and december/2009. The results showed that one quarter of patients who developed surgical site infection in the hospital needed a new hospitalization. The mean age of subjects was 57.2 years; more than a half had diabetes mellitus, and a third, hypertension, however, the anesthetic risk for the majority was classified as low. The infections occurred more frequently in clean and potentially contaminated surgeries and the medical specialties with higher number of patients readmitted was orthopedic, with more than half of surgeries related to the correction of fractures, and general surgery, where the hernia and gallbladder surgery predominated. Half of the subjects received biomaterial in its proceedings. The surgical and hospitalization times did not differ from others studies in the literature. Virtually all patients received antibiotic prophylaxis for indications mentioned in the protocol. Early signs and symptoms appeared after an average of 33.2 days, and half of the patients were diagnosed within the first 30 days postoperatively. The classification of infections showed that half of the patients had infection of organs and cavities and 60% were required surgical interventions, in addition to antimicrobial treatment. Staphylococcus aureus predominated in orthopedic procedures and Escherichia coli in general surgery, both with multi-resistance profile below the results presented in other studies. The study concludes that the post-discharge surveillance and monitoring of readmission rates contribute to the scaling of the problem and define specific actions for its control.

Estudo de desenvolvimento metodológico que teve por finalidade disponibilizar um sistema de avaliação de Programas de Prevenção e Controle de Infecção Hospitalar (PCIH), após validação das propriedades de medidas, que possibilite aplicação prática para diagnósticos situacionais, cujos resultados subsidiem tanto melhorias na área quanto informações concretas à sociedade sobre a qualidade desses PCIH em cada instituição de saúde. Para tal, previamente, foram construídos e realizada validação de conteúdo de quatro indicadores: 1- (PCET) Estrutura Técnico-operacional do PCIH; 2- (PCDO) Diretrizes Operacionais de Controle e Prevenção de IH; 3- (PCVE) Sistema de Vigilância Epidemiológica de IH; 4- (PCCP) Atividades de Controle e Prevenção de IH. Os objetivos específicos foram: realizar validação (construto e discriminante) e testar confiabilidade (consistência interna); realizar diagnóstico parcial de conformidade dos PCIH em serviços de saúde do Município de São Paulo. Os indicadores de PCIH foram aplicados em 50 instituições de saúde, que aceitaram voluntariamente participar do estudo. As características referentes ao perfil do hospital e os escores dos indicadores foram descritos por meio de estatística descritiva. A consistência interna foi analisada usando o coeficiente de Cronbach. A análise da validade discriminante foi realizada comparando-se os escores dos indicadores entre dois grupos de hospitais, (possuem algum tipo de certificação versus não possuem certificação em qualidade). E para a análise da validade de construto foi utilizada a Análise Fatorial Exploratória com matriz de correlação tetracórica. Os indicadores 1-PCET e 3-PCVE variaram pouco, com quase 100% de conformidade em toda a amostra, já os indicadores 2-PCDO e 4- PCCP apresentaram boa consistência interna com variação de 0,67 a 0,80. A validade discriminante desses indicadores indicou médias dos escores de conformidade superiores e com significância estatística no grupo de instituições com processos de qualificação ou acreditação em saúde. Na validação de construto foi possível diferenciar e identificar 2 dimensões para PCDO (fator 1- recomendações para prevenção de IH e fator 2 recomendações para padronização de procedimentos de profilaxia), com boa correlação dos itens que o compõe, o mesmo ocorrendo para PCCP (fator 1 interface com unidades de tratamento e fator 2 interface com unidades de apoio). N avaliação parcial de conformidade do PCIH todos os indicadores, com exceção do 4 PCCP, que variou de 9,5% a 100%, apresentaram escores > 90%,o que mostra que os hospitais participantes possuem um bom nível de qualidade dos PCIH instituídos, com médias superiores nas instituições com processos de qualificação da assistência. Com esse estudo foi possível validar as propriedades de medidas dos indicadores de PCIH e disponibilizar um instrumento factível como ferramenta de avaliação de PCIH de forma ética e científica para diagnóstico de qualidade na área. / The objective of this study was to develop methodology that could ultimately be made available as a system for assessing Hospital Infection Control and Prevention Programs (HICPP), after validation of its measurement properties, which enable practical application in situational diagnoses, whose results provide both improvements in the area and reliable information about the quality of these HICPPs in each healthcare institution. To do so, the content of four indicators was constructed and validated beforehand: 1- (PCET) Technical-operational structure of the HICPP; 2- (PCDO) Operating Guidelines for Control and Prevention of HI; 3- (PCVE) Epidemiological Surveillance System for HI; 4- (PCCP) HI Prevention and Control Activities. The specific objectives were: fully validate (construct and discriminant) and test the reliability (internal consistency) of measurement properties; carry out a partial diagnosis of conformity of the HICPPs in São Paulo Municipal Healthcare Facilities. The HICPP indicators were applied in 50 healthcare institutions, which participated voluntarily in the study. The hospital profile characteristics and the scores of the indicators were described using descriptive statistics. Internal consistency was analyzed using the Cronbach coeficient; the analysis of discriminant validity was carried out by comparing the scores of the indicators between the two groups of hospitals, (those which had some type of quality certification versus those which did not) and exploratory factor analysis with a tetrachoric correlation matrix was used to analyze the validity of the construct. The indicators 1-PCET and 3-PCVE varied little, with almost 100% conformity throughout the sample, whereas the indicators 2-PCDO and 4- PCCP presented good internal consistency with a variation of 0.67 to 0.80; discriminant validity of these indicators showed higher average scores of conformity and were statistically significant in the group of institutions with certification or accreditation; in the validation of the construct it was possible to differentiate and determine 2 dimensions for PCDO (factor 1- recommendations for prevention of HI and factor 2 recommendations for the standardization of prophylaxis procedures), with good correlation of the units of analysis that composed it. The same occurred for PCCP (factor 1 interface with treatment units and factor 2 interface with support units). All of the indicators, with the exception of 4- PCCP, which ranged from 9.5% to 100%, presented scores of > 90%, which show that the HICPPs of participating hospitals have a good standard of quality, with higher average scores in the institutions with certification or accreditation. This study enabled the validation of the measurement properties of the HICPP indicators and produced a practicable HICPP assessment tool in an ethical and scientific manner for diagnosis of quality in this area.

L’infection urinaire (IU) est un problème majeur de Santé publique. La cystite aiguë touchant principalement les femmes est la plus fréquente des IU. La bactérie la plus fréquemment isolée au cours de ces IU est Escherichia coli. Une des particularités de la cystite est sa propension à récidiver. Le traitement préconisé pour ces infections est la prise d’antibiotiques, qui peut être fréquente en cas de cystites récidivantes. C’est dans ce contexte que de nouvelles stratégies doivent être développées afin de prévenir et traiter les IU récidivantes. Parmi ces différentes stratégies, l’utilisation de produits naturels tels que la canneberge (Vaccinium macrocarpon) apparaît comme prometteuse. En effet, des études précédentes ont montré que la canneberge a un effet négatif sur l’adhésion des bactéries aux cellules superficielles de l’épithélium vésical facilitant l’élimination des bactéries par le flux urinaire. Cette activité est portée par la proanthocyanidine de type A (PAC-A). D’autre part, une étude menée par notre équipe a montré que l’effet de la canneberge sur l’adhésion et la virulence de souches d’E. coli uropathogènes pouvait être potentialisé par l’ajout d’un autre composé naturel : la propolis. Depuis l’Antiquité ses propriétés anti-bactériennes sont reconnues et des études plus récentes ont démontré son impact sur des bactéries à Gram positif mais également sur deux bactéries à Gram négatif : E. coli et Pseudomonas aeruginosa. Ce travail de thèse a permis : i) de décrire l’impact de la canneberge, de la propolis et de leur association sur le transcriptome d’une souche clinique d’E. coli uropathogène (G50). Cette analyse transcriptomique a montré que la canneberge entrainait une sous-expression de gènes liés à l’adhésion, mais également de gènes liés à la mobilité et à la formation de biofilm. En revanche, la canneberge augmentait l’expression des gènes liés au métabolisme du fer ainsi qu’à la réponse au stress. Ces effets étaient potentialisés par l’ajout de la propolis. En parallèle, des tests phénotypiques menés sur une collection de souches d’E. coli uropathogènes sur la mobilité et la formation de biofilm ont confirmé les résultats précédents ; ii) de développer un test, basé sur les précédents travaux de transcriptomique, permettant une évaluation standardisée de l’effet de la PAC-A sur E. coli, indépendamment de sa concentration car il n’existe pas de techniques standardisées pour doser cette molécule. C’est ainsi que 4 gènes (tsr, ftnA, fecB, feoB) ont été sélectionnés, le suivi de leur expression permettant une mesure de l’activité anti-bactérienne de la canneberge; iii) de mesurer l’effet potentialisateur de la propolis sur l’activité des antibiotiques utilisés dans le traitement des IU. C’est ainsi qu’il a été montré que l’ajout de la propolis permettait d’augmenter l’activité bactéricide des antibiotiques testés et de diminuer les concentrations minimales inhibitrices de ces antibiotiques. / Urinary Tract Infection (UTI) is a major problem of public health. Acute cystitis which touches mostly women is the most common form of UTI. The bacteria which is mostly isolated in an UTI is Escherichia coli. A particularity of cystitis is to come back. In this context news strategies have to be developed to prevent and cure recurrent UTI. One of these strategies is the utilization of natural products like the cranberry (Vaccinium macrocarpon) which is promising. Indeed, previous studies showed the negative impact of cranberry on the adhesion of bacteria on the superficial cells of bladder which help the elimination of bacteria by the urinary flux. This activity is carried by the type A proanthocyanidin (PAC-A). Moreover, a study lead by our team has demonstrated an improvement of the activity of cranberry on the adhesion and the virulence of uropathogenic E. coli (UPEC) by another natural product: the propolis. Since Antiquity its antibacterial activities have been recognize and more recent studies have demonstrated its impact of Gram positive bacteria and also on two Gram negative bacteria: E. coli and Pseudomonas aeruginosa. This thesis has allowed for: i) the description of the impact of cranberry, propolis and its association on the transcriptome of a clinical strain of UPEC (G50). This transcriptomic analyze have shown that the cranberry down regulated genes linked to the adhesion and also genes linked to the motility and biofilm formation. However the cranberry up regulated genes linked to the iron metabolism and the stress response. These effects are improve by the addition of propolis. Concurrently phenotypics tests have been conducted on a collection of UPEC on the motility and the biofilm formation and they confirmed the previous results; ii) the development of a test, based on our transcriptomic results, enable to performed a standardized evaluation of the impact of PAC-A on E. coli, independently of its concentration. Indeed, this molecule cannot be measure in a standard way. Four genes have been selected (tsr, ftnA, fecB, feoB), the monitoring of their expression allow us to measure the anti-bacterial efficiency of the cranberry; iii) the measurement of the potential effect of the propolis of the antibiotic’s activities used to treat UTIs. Thus it have been observed that the addition of propolis improve the bactericide activity of the antibiotics tested and reduces the minimal inhibitory concentration of these antibiotics.

Mycobacterium tuberculosis est la bactérie responsable de la tuberculose (TB), une infection pulmonaire grave. La TB est un problème majeur de santé publique mondial ; un tiers de la population mondiale est porteur de M. tuberculosis et l'émergence de formes de TB résistantes aux antibiotiques confirme la nécessité de développer de nouvelles approches thérapeutiques. Pour atteindre cet objectif, il est crucial de mieux comprendre les mécanismes infectieux de M. tuberculosis. L'équipe de C. Guilhot s'intéresse aux lipides de l'enveloppe bactérienne impliqués dans la virulence de M. tuberculosis, en particulier lors de l'interaction du bacille avec les macrophages de l'hôte. Un type de lipide en particulier s'est démarqué pour son rôle crucial dans cette interaction : les Dimycocérosates de Phthiocérol (DIM). Malgré leur importance, le mécanisme d'action moléculaire des DIM n'est toujours pas connu. Les objectifs de ma thèse ont été 1) d'étudier le rôle des DIM dans le trafic intracellulaire de M. tuberculosis au sein du macrophage, et 2) de comprendre les mécanismes d'action des DIM dans la virulence. Par une combinaison de méthodes biologiques et biophysiques, nous avons montré que les DIM contribuent à l'induction de la rupture du phagosome et de l'apoptose du macrophage infecté par M. tuberculosis, en collaboration avec un autre facteur majeur de virulence bactérien, ESAT-6. Au niveau moléculaire, nous avons confirmé que les DIM sont transférés dans la membrane du macrophage au contact avec la bactérie, et y induisent une rigidification membranaire locale. Nous avons aussi montré que les DIM dans une membrane sont capables de potentialiser l'activité membranolytique d'ESAT-6 et d'autre(s) facteur(s) encore non identifiés. Les DIM ont un rôle pléiotropique dans l'interaction de M. tuberculosis avec le macrophage. Leur mécanisme d'action impliquerait des modifications des propriétés biophysiques membranaires, modifiant l'activité des effecteurs membranaires bactériens et potentiellement de l'hôte. Ces travaux ouvrent la voie à l'étude du mécanisme d'action d'autres lipides de virulence dont certains pourraient également s'insérer dans la membrane du macrophage. / Mycobacterium tuberculosis is the bacterium responsible for tuberculosis (TB), a severe respiratory disease. TB is a major public health threat; one third of the world's population is latently infected by M. tuberculosis, and the emergence of antibiotic-resistant forms of TB confirms that there is a need to develop new therapeutic approaches to control the spread of TB. However, in order to attain that goal, it is crucial to decipher the infectious mechanisms of M. tuberculosis. Research in the team of C. Guilhot focuses on lipids from the envelope of M. tuberculosis which are involved in virulence, in particular those implicated in the interaction of the bacteria with host macrophages. One of these lipids stands out for its crucial role in this interaction: Phthiocerol Dimycocerosate (DIM). Despite its importance, the molecular mechanism of action of DIM is still unknown. The objectives of my PhD were 1) to study the role of DIM in the intracellular trafficking of M. tuberculosis in macrophages, and 2) to decipher the molecular mechanism of action of DIM. By a combination of biological and biophysical techniques, we showed that DIM contributes to phagosomal rupture and induction of apoptosis in macrophages infected with M. tuberculosis, in collaboration with another major virulence factor: ESAT-6. At the molecular level, we confirmed that DIM is transferred to the membrane of the macrophage on contact with M. tuberculosis and induces a local membrane rigidification around the point of contact with the bacterium. We observed that DIM incorporated in membranes is able to promote the membranolytic activity of ESAT-6, and other yet unidentified bacterial factor(s). DIM has a pleiotropic role in the interaction between M. tuberculosis and macrophages, presumably through alterations of the membrane's biophysical properties that influence the activity of membrane effectors from both the bacteria and the host. Thus, this work paves the way for the study of the mechanisms of action of other virulence lipids, some of which could also be inserted in the macrophage membrane.